Note: these are in order of presentation during the week.
Friis, E 1
1 Department of Palaeobotany, Swedish Museum of
Natural History, Stockholm, Sweden
A multitude of small, charcoalified flowers, fruits, seeds and other reproductive structures have been retrieved from sediments of Cretaceous age. The fossils are highly informative, in some cases preserving astonishing morphological and anatomical details. Excellent preservation, together with the application of advanced techniques such as synchrotron-based X-ray microtomography, has resulted in unexpected insights into the diversification and reproductive biology of early angiosperms. Documentation of extinct lineages with character combinations unknown in extant angiosperms are particularly interesting for analyses of character evolution and may help bridge some of the gaps in the phylogenetic tree of extant angiosperms. The fossil flowers of the newly described Canrightia resinifera provide an example of an Early Cretaceous angiosperm with intriguing floral structure that combines features of several extant magnoliid lineages and it is suggested that
Canrightia may be a link between Chloranthaceae and
Piperales close to the root of the eumagnoliid tree.
Phylogenetic and structural signals from the fossil record also shed new light on Darwin's mystery: 'The rapid development as far as we can judge of all the higher plants within recent geological times is an abominable mystery'. The fossils indicate that the major diversification of angiosperms was preceded by a longer, more gradual radiation. Great changes in floristic diversity and distribution from the Early to Late
Cretaceous and between Cretaceous and Cenozoic are evident. Most significant is the high diversity of Early
Cretaceous fossils related to ANITA-grade angiosperms,
Chloranthaceae and alismatalean monocots indicating predominance of herbs, small shrubs and aquatic plants in the early history of angiosperms. These probably had restricted ecological ranges, which may explain the paucity of angiosperm leaves and pollen in strata representing the earliest phases of angiosperm evolution.
Eumagnoliids and eudicots were also established early, but both groups are rare at this level of angiosperm evolution, and it is significant that no core eudicots have yet been encountered among Early Cretaceous angiosperms. However, around the mid-Cretaceous eudicot diversity increased rapidly, particularly through a considerable radiation of various rosid lineages (e.g.,
Fagales) and basal lineages of asterid angiosperms (e.g.,
Cornales and Ericales). Eumagnoliids also show marked diversification at this time with particular development of many taxa related to Laurales. These phylogenetic changes greatly affected vegetation structure resulting in a predominance of woody angiosperms in the Late
Cretaceous floras.
1
Fischhoff, DA 1
Monsanto Company, St Louis, Missouri, USA
Genetic improvement of crop plants in the past two decades primarily focused on the implementation of
DNA marker technologies in plant breeding programs and on the first generation of crop biotechnology traits that provided weed control via herbicide tolerance and systems for insect resistance. Traits developed through both biotechnology and plant breeding efforts that protect yield potential against pests and disease will continue to be important, especially when considering possible climate-related changes in geographical range and reproductive capabilities of weeds and insects. Future generations of pest control traits are being developed through the application of new technologies including protein engineering and RNAi to offer growers products with additional modes of action and ever-improving efficacy against pests. In addition, significant research efforts in crop biotechnology now focus on enhancements to complex traits such as intrinsic yield potential, nutrient utilization and abiotic stress mitigation, which when coupled with similar efforts to improve these traits via advanced plant breeding techniques and agronomic practices, could represent a step-change in crop yield performance. Increasing yields while at the same time significantly decreasing the key resources (water, land and energy) required to produce each unit of output is one of the most important challenges facing agriculture. Leveraging advanced enabling technologies such as high-throughput genome sequencing, functional genomics, and systems biology in both plant breeding and biotechnology disciplines will be required to help deliver the next generation of traits in agricultural crops.
Oostermeijer, G 1
1 Institute for Biodiversity and Ecosystem Dynamics,
University of Amsterdam, The Netherlands
Already in the 1980s, pioneer conservation biologists discussed the possibly important role of genetics in the viability of small and isolated populations. Since then, an enormous amount of research has shown that such populations are experiencing loss of genetic variation through drift and inbreeding, and that gene flow is often insufficient to compensate for that. However, when it comes to the question how detrimental this is to their viability, we are still not able to present very many empirical studies that clearly demonstrate this. This is largely due to our failure to effectively integrate genetics with other important components of plant population viability, such as demography and reproductive biology.
Nevertheless, the studies performed on each of these components separately suggest that there are major changes in each of them. The still painfully scarce studies that have attempted to integrate them into single PVA's
1

show that demography, reproduction and genetics act synergistically, and cause the often mentioned but rarely demonstrated extinction vortex. As a result of habitat destruction and fragmentation, many plant populations are genetically depauperate, and show reduced viability that cannot be alleviated by habitat management and restoration alone and requires additional genetic rescue measures. Our knowledge of the implications of outbreeding depression after genetic rescue is still insufficient to provide conservation managers with scientifically sound advice. The rapid developments in ecogenomics will provide exciting new avenues of implementing genetics into plant conservation.
Nevertheless, we still need to combine these new approaches with appropriate field experiments in order to obtain the best answers to still urgent questions. To better implement genetics into practical conservation, researchers need to (i) pay more attention to translating their scientific results, and (ii) focus on the specific questions of conservation managers.
Prusinkiewicz, P 1
1 University of Calgary, Calgary, Canada
Branching architecture is a conspicuous feature of most plants . It is also difficult to conceptualize, as it is not coded directly in the plant genotype, but emerges from a hierarchy of morphogenetic processes in an indirect and often non-intuitive way. As early as the 1960s and 1970s, computer models were introduced to investigate the relations between local control of development and the global branching form of plants . Over the last decade, models rooted in molecular-level data and hypotheses have offered further insights. At the basis of these models lies a postulated feedback between the concentrations and flow of auxin, and the distribution of auxin transporters, in particular PIN1 proteins . Operating in the shoot apical meristems, this feedback can explain the spatial arrangement of buds around their supporting axes
(phyllotaxis). Operating along the plant axes, a related feedback mechanism explains which buds will eventually develop into branches, and at which points of time (bud activation). Of particular interest are strategies that plants employ to avoid overcrowding if the branching processes repeat and several orders of branches are formed . One strategy, employed by herbaceous plants and inflorescences, is to gradually reduce the size of branches, producing fractal patterns . Another strategy, widely employed by trees, is to avoid overcrowding through competition between buds and branches for light and space . Recent models show that a wide variety of temperate-climate tree architectures results from different biases in this competition . Although many gaps in our knowledge remain, the link between molecular-level processes and the branching architecture of herbaceous plants and trees begins to appear.
Higashiyama, T 1
1 Nagoya University, ERATO Higashiyama Live-Holonics
Project, Japan
In the fertilization process of flowering plants, gametophytic interactions between the pollen tube and the embryo sac occur deeply inside the pistil of the flower. Due to the inaccessibility, it still remains unclear how gametophytic cells communicate to achieve double fertilization. We developed the in vitro Torenia system, whereby pollen tubes growing through a cut style are attracted to a protruding embryo sac and cause double fertilization. By using this system, the synergid cell was shown to emit some diffusible attractant(s) (Higashiyama et al., Science, 2001), which had been sought for more than 140 years. We investigated genes expressed in the synergid cell of Torenia , by collecting isolated synergid cells. We found that cysteine-rich peptides (CRPs) were abundantly expressed in the synergid cell. Among the
CRPs, at least two defensin-like polypeptides, named as
LUREs, showed strong activity to attract pollen tubes.
By developing a laser-assisted thermal-expansion microinjector, LUREs were finally identified as attractants derived from the synergid cell (Okuda et al.,
Nature, 2009). One of our goals is to clarify the mechanism of directional control of pollen-tube growth by LUREs. In this talk, I will show our recent progress, including results based on visualization of LURE molecules as well as pollen tube guidance of Arabidopsis in vivo. On the other hand, we developed a method for live cell imaging of fertilization in Arabidopsis . I will also discuss the mechanism of double fertilization based on the direct observation of fertilization processes
(Hamamura et al., in press).
Myburg, AA
J
L
5
6 , Rouzé, P 6
U 7,8
Bristow, J 8
1
, Schmutz, J
, Grattapaglia, D
5 , Hefer, CA
, Goodstein, D 7,8
1
2
, Van De Peer, Y 6
, Tuskan, GA
, Pappas Jr, GA
, Hayes, R 7,8
, Rokshar, DS 7,8 , Barry, K
, The Eucalyptus Genome Network
,
9
4
2,3
, Jenkins,
, Sterck,
Hellsten,
8 ,
1 Dept of Genetics, Forestry and Agricultural
Biotechnology Institute (FABI), University of Pretoria,
2 Pretoria, South Africa; Plant Genetics Laboratory,
EMBRAPA Genetic Resources and Biotechnology -
3 Genomic Sciences Program, EPqB, Brasilia, Brazil;
Universidade Católica de Brasília - SGAN, Brasília,
Brazil; 4 Environmental Sciences Division, Oak Ridge
5 National Laboratory, Oak Ridge, USA; HudsonAlpha
Genome Sequencing Center, Huntsville, USA; 6 Dept of
7
Plant Systems Biology, Ghent University, Gent, Belgium;
Center For Integrative Genomics, Dept of Molecular and Cell Biology, University of California, Berkeley,
USA; 8 DOE Joint Genome Institute, Walnut Creek, USA;
9 www.eucagen.org
Almost all of the more than 700 known species of
Eucalyptus L'Hér. (Myrtaceae) are endemic to Australia.
They are a dominant component of the flora of
2

Australasia occupying 79% of the native forest estate.
Worldwide, eucalypt species and hybrids constitute the largest hardwood fibre crop with over 20 million hectares of plantations grown primarily for timber, pulp and paper. Fast-growing eucalypts are also a promising source of lignocellulosic biomass for energy production.
The Eucalyptus genome sequence, the first for an
Australian plant, will offer important benefits for ecological and evolutionary biology studies, as well as for tree breeding and biotechnology. Over the past three years, the Department of Energy (DOE) Joint Genome
Institute (JGI) has performed whole-genome (8X) shotgun sequencing for Eucalyptus grandis (est. genome size 640 Mbp, n = 11), combining 7.7 million Sanger reads from plasmid, fosmid and BAC libraries of a partially inbred (S1), but fully viable 17-year-old tree,
BRASUZ1. Due to high residual heterozygosity, initial genome assembly resulted in approximately 25% of the assembly occurring in two haplotypes of 3–4X coverage, while the remainder of the genome assembled into a single haplotype of 6–7X coverage. High-density linkage maps with >2000 DArT and microsatellite markers guided the subsequent assembly of 11 large chromosome
(pseudomolecule) scaffolds containing 88% (605 Mbp) of the draft assembly. Similarity searches with 1.6 million E. grandis ESTs suggested that a large proportion
(96%) of expressed gene loci are located in the mapped chromosome assemblies. Ab initio and homology-based annotation performed in parallel efforts at the JGI and the
University of Ghent was further supported by over 4 million 454-FLX-Titanium ESTs produced by JGI, as well as Sanger and Illumina EST data provided by collaborators. The annotations produced by the two groups revealed that the 11 chromosome scaffolds contain more than 90% (41,201 – JGI, 43,219 – UGent) of the predicted protein-coding loci, of which more than
70% have EST support. A preliminary analysis of genome duplication performed at UGent suggested that, in addition to the ancient hexaploidization event shared by Rosids and Asterids, the Eucalyptus genome most likely contains one more recent duplication event. The two genome annotations were released in public databases (www.phytozome.net, http://bioinformatics.psb.ugent.be/webtools/bogas/) in
Jan 2011 . The E. grandis genome sequence will be the first reference for the Rosid order Myrtales and will therefore be informative for comparative genomic studies within the Eudicots .
Quail, P 1
1 Plant and Microbial Biology, UC Berkeley, and Plant
Gene Expression Center, Albany, USA
A central goal of current phytochrome (phy) research is to define the cellular, molecular and biochemical mechanisms involved in the primary steps of the lighttriggered, intra-cellular transduction process utilized by this photoreceptor family (phyA to phyE). The present paradigm charting this signaling pathway asserts that transduction involves rapid translocation of the lightactivated photoreceptor molecule from the cytoplasm to the nucleus, where it interacts physically with a subset of members of the bHLH transcription factor family, termed
Phytochrome-Interacting Factors (PIFs), inducing transcriptional responses in target genes. We have shown that a quadruple pif mutant (pif1pif3pif4pif5 abbreviated pifq) exhibits a constitutively photomorphogenic (cop)like phenotype in completely dark-grown seedlings, indicating that these transcription factors collectively repress photomorphogenesis in post-germinative darkness, and that photoactivated phy reverses this repression by inducing rapid degradation of the PIF molecules upon initial exposure to light. This process involves rapid, phy-induced phosphorylation of the interacting bHLH protein, followed by degradation via the ubiquitin proteasome system. Using random mutagenesis, we have identified a surface-exposed binding site for the PIFs in the light-sensing knot region of the photoreceptor, that is necessary for normal phy signaling in inducing seedling deetiolation, suggesting that this site is integral to the biochemical signal transfer process from the activated photoreceptor to target proteins in the cell. Microscopic examination of the pifq mutant indicates accelerated oil body mobilization and chloroplast differentiation in dark-grown seedlings, phenocopying the normal light-induced deetiolation process in the wild-type. Genome-wide expression profiling of the pifq seedlings shows robust derepression of a broad array of nuclear genes encoding chloroplasttargeted proteins in sustained darkness, consistent with the visible and cellular cop-like phenotypes. Collectively, the data support the notion that the PIF subfamily of bHLH transcription factors function constitutively to promote skotomorphogenic development in seedlings emerging from buried seed, repressing premature photomorphogenic development in subterranean darkness, until this repression is relieved by proteolytic degradation of the PIFs upon photoactivation of the phy system by initial exposure to light at the soil surface . The transcriptome analysis has identified a subset of genes that both are regulated by the PIFs in darkness in the pifq mutant, and are rapidly responsive to light in the wildtype . These genes are thus potential direct targets of phy regulation via the PIF transcription factors . Recent evidence indicates that the PIFs may have a role in multiple signaling pathways in addition to light, thereby suggesting that they function as components of a central cellular signaling hub.
Wendel, JF 1
1 Dept of Ecology, Evolution, & Organismal Biology,
Iowa State University, USA
Increasingly powerful technologies arebeingused to study the ecology and evolution of polyploids in many plant groups, resulting in dramatic discoveriesof novel genomic interactions and processes. Gossypium includes classic allopolyploids arising from a biological reunion1-
2 MYAof divergent diploids from different hemispheres.
This serendipitous merger generated a spectrum of genomic responses, including gene silencing, intergenomic gene conversion, and genome-wide
3

disruption and reconciliation of ancestral gene expression patterns. Using several microarray platformsand other technologies, we are studying transcriptional changes in synthetic and natural Gossypium allopolyploids and reconstructed F1 and polyploid hybrids, using differing tissues and genetic backgrounds. Allopolyploid formation induces massive alteration in gene expression and complex transcriptomicresponses, including genomic dominance and novel (transgressive) expression patterns.
Using microarrays and sequencing protocols that distinguish transcript levels for each member
(homoeolog) of each duplicated gene,we show that allopolyploidization entails significant homoeolog expression modulation that is temporally partitioned into alterations arising immediately as a consequence of genomic merger and secondarily as a result of long-term evolutionary transformations in duplicate gene expression.About one fourth of homoeolog expression biases occur instantaneously with hybridization, with the remaining arising from long-term evolutionary forces such as duplicate gene neofunctionalization and subfunctionalization. Expression is biased toward the paternal D-genome, demonstrating an overall unequal contribution of two genomes to the transcriptome. We are exploring gene expression during cotton fiber development and evolution, taking advantage of a wellestablished phylogenetic framework and the unique opportunity offered by the existence of multiple, parallel domestications of different wild progenitors by aboriginal peoples in both Africa–Asia and
Mesoamerica. The fibertranscriptome is extraordinarily complex, with homoeolog expression varying widely even at the level of development and maturation of a single cell. Most homoeolog expression bias reflects polyploidy rather than domestication, but domestication has increased expression bias in fibers towards the Dgenome.We are exploring the functional consequences of gene duplicationin cotton and the possibility of novel gene recruitment following genome doubling.This work provides insights into the genetic architecture underlying the evolution of morphology, as well as the potential evolutionary significance of genome doubling.
Crisp, M 2
1 This is the 2011 Nancy Burbidge Memorial Lecture of the Australasian Systematic Botany Society
2 The Australian National University, Canberra, Australia
Australia has changed from a rainforest-covered
Gondwanan landscape to an arid island-continent dominated by sclerophyll plants such as eucalypts and acacias. What were the key evolutionary and ecological drivers of this transformation? By integrating the fossil record with molecular phylogenetics, we can help reconstruct the time line of evolutionary history and environmental changes can be identified by their historical signatures in phylogenies . Not surprisingly, most lineages can be traced to ancestors in wet habitats .
However, reconstructions of fire-adaptive traits in
Myrtaceae and Proteaceae suggest that flammable sclerophyll communities might have existed in the early
Palaeocene, 60 Myr ago. After Australia separated from
Gondwana 30 Myr ago, the climate became more seasonal and the monsoon tropical flora was assembled both by in situ adaptation of Gondwanan lineages and from immigrants from the north as Australia drifted towards southeast Asia . Comparisons of divergence times among multiple pairs of lineages on either side of the arid Nullarbor Plain barrier support the hypothesis that mid-Miocene aridification (15 Myr ago) isolated the temperate sclerophyll floras of southwestern and southeastern Australia . The largest and youngest biome, the arid zone, is thought to contain a mix of young taxa adapted from the mesic near-coastal regions, Asian immigrants and relicts of the ancestral rainforest, clinging to aquifers in the central Australian ranges .
Surprisingly, one 'relict' group, the Livistona fan palms, have turned out to be very recent immigrants . These findings have raised more questions than they have answered and I will discuss some of these new challenges .
Somerville, C 1 , Gu, Y
A 1 , Anderson, C 1
2 , Chen, S
, Sorek, N 1
1 , Wallace, I 1 , Carroll,
, Paredez, A 1
1 University of California, Berkeley, USA;
State University, USA
2 Pennsylvania
Cellulose is thought to be the most abundant component of terrestrial biomass because of its role as the major structural element of plant cell walls. Cellulose is also a component of many commodities such as lumber, paper, textiles and animal fodder. Thus, it is surprising that relatively little is known about how the properties of cellulose are controlled during synthesis. In 2006 we reported that it was possible to observe what appeared to be individual cellulose synthase complexes synthesizing cellulose in live cells using GFP-labeled complexes and spinning disc confocal microscopy. This method of observing the process has been used by us and others to observe the effects of genetic and environmental perturbations of the process and to infer some aspects of the molecular details of the process . This general approach has shown that cellulose synthase is staged in the Golgi from where it is secreted in vesicles to the plasma membrane. At the plasma membrane the complex is activated and associates with the cortical cytoskeleton which plays a role in orienting the deposition of cellulose fibrils in a pattern that is regulated dynamically in response to developmental and environmental cues .
Recent results indicate that phosphorylation of the cellulose synthase complex participates in some aspect of the interaction between the cellulose synthase complex and cortical microtubules in primary cell walls. Several of the kinases that act on cellulose synthase have recently been discovered and may open up new opportunities to identify the signal transduction pathways that link the process to cellular functions . Additionally, a number of other proteins that have been implicated in cellulose synthesis by mutant studies have been shown to alter the kinetics of the process and several appear to colocalize with the cellulose synthase complex . The complexity revealed by the study of the many factors that affect the overall process suggest that cellulose synthesis is a much
4

more complex process than merely polymerizing glucose. We have come to think of it as a molecular process that is similar in complexity to transcription or translation; resolving the details of the process will require a similar level of ingenuity as was required to elucidate those processes
Asymmetries of DNA methyltransferases activity between male and female gametes cause imprinting
Arabidopsis
Berger, F 1 , Thiet, VM 1 , Jullien, PE 1
1 Temasek Lifescience Laboratory, Singapore
Imprinted genes are expressed predominantly by one parental allele. In plants, most imprinted genes are expressed in endosperm, the tissue that nurtures embryo development inside the seed. The endosperm derives from the fertilized central cell. Imprinted expression relies on DNA demethylation occurring in the central cell but not in sperm cells. After fertilization, the asymmetry of DNA methylation acquired at imprinted loci during gametogenesis is inherited to endosperm where a transcriptionally active maternal allele coexists with a silenced paternal allele, resulting in an imprinted status.
Demethylation in the central cell relies both on the inactivation of MET1 transcription and the expression of the DNA demethylase DEMETER. Hence imprinting has so far been linked exclusively to cytosine methylation on
CG contexts, which depends on MET1 activity. The
RNA dependent DNA methylation (RdDM) pathway causes de novo methylation of cytosine residues in any context and is involved in silencing transposons, participates to the maintenance of constitutive heterochromatin around centromeres, as well as the silencing of transgenes. We report that the RdDM pathway is active only in male gametes but not in the central cell. The inactivation of the RdDM pathway in the central cell relies on specific transcriptional activation of the major player of this pathway by the retinoblastoma pathway in a manner similar to that reported for MET1. The asymmetry of RdDM activity is sufficient to cause imprinting of several loci. We present the case of an imprinted gene controlled by the RdDM pathway and extrapolate further generalization of the mechanism.
A credible molecular framework for small RNAbased transgenerational effects in plants
Brosnan, C 1 , Dunoyer, P 1 , Voinnet, O 1
1 Swiss Federal Institute of Technology, Zurich,
Switzerland
We have recently demonstrated that endogenous and exogenous small RNAs move from cell–to–cell and over long distances in plants. Hundreds of small RNAgenerating loci are found in the Arabidopsis genome, and we have shown that their expression, processing and activity may be influenced by specific stresses and may also vary extensively from one plant ecotype to another.
Because a significant fraction of mobile small RNAs has the potential to induce DNA methylation and chromatin modification at homologous loci, a model emerges whereby somatic integration of environmental cues might be converted into large bulks of mobile genetic information that could ultimately pattern gene expression into the meristems, the new growth and, possibly, the gametophytes. This process may not only adapt dynamically the new growth to the changing environment, but may also have long-term transgenerational consequences owing to the action of plant maintenance DNA methylases and the absence of resetting in these organisms. It may also explain the formation of ‘spontaneous epialleles’ arising under specific environmental conditions. I will present a series of molecular data and striking observations, which, collectively, constitute a credible framework for this proposed transgenerational small RNA-based memory in plants.
Heterochromatin reprogramming by small RNA regulates germ cell fate and transposon silencing in polyploid hybrids
Martienssen, R 1
K 1 , VanEx, F
Ernst, E 1
1
, Tanurdzic, M
, Auer, P 3
, Vielle-Calzada, J
1 , Finigan, P
, Vaughn, M
4 , Doerge, R 3
1
1 , Creasey,
, Slotkin, K 1,2 ,
1 Cold Spring Harbor Laboratory, Cold Spring Harbor,
NY, USA; 2 Ohio State University, Columbus, OH, USA;
3 Dept of Statistics, Purdue University, West Lafayette,
IN, USA; 4 CINVESTAV, Irapuato, Mexico
In many plants, including Arabidopsis , hybrids between species and subspecies encounter post-fertilization barriers in which hybrid seed fail to develop, or else give rise to infertile progeny . In Arabidopsis , some of these barriers are sensitive to ploidy and to the epigenetic status of donor and recipient genomes . Recently, a role has been proposed for heterochromatin in reprogramming events that occur in reproductive cells, as well as in the embryo and endosperm after fertilization . 21nt epigenetically activated small interfering RNA
(easiRNA) from transposable elements accumulate in cultured cells and in pollen, and are translocated from the surrounding pollen grain into the sperm, while in the maturing seed 24nt siRNA are primarily maternal in origin. Thus maternal and paternal genomes likely contribute differing small RNA to the zygote and to the endosperm. As heterochromatic sequences also differ radically between and within species, small RNA sequences will diverge in hybrids . If transposable elements in the seed are not targeted by small RNA from the pollen, or vice versa, this could lead to hybrid seed failure, in a mechanism reminiscent of hybrid dysgenesis.
Unexpectedly, mutants in the easiRNA pathway lead to specification of diploid functional megaspores from somatic cells in the ovule, reminiscent of apospory in other species, as well as meiotic defects. Thus heterochromatin reprogramming may play a role in apomixis, and may utilize a similar mechanism .
5

The nature of species boundaries in plants
Rieseberg, L
Scasitelli, M 1 ,
1 , Kane, N 1
Strasburg, J 2
, Andrew, R 1 , Renaut, S 1
1 University of British Columbia, Canada; 2
,
Indiana
University, Bloomington, USA
Despite numerous reports of hybridization in natural populations of plants and animals, hybridizing species typically remain morphologically distinct due to the action of multiple reproductive barriers. Less is known about pattern of molecular genetic differences between hybridizing taxa. In a recent monograph on speciation,
Jerry Coyne and Allen Orr argue 'much of the genome cannot move between species because it is linked to divergently selected alleles'. However, recent population genomic studies suggest that reproductive barriers may be more porous than conventionally believed, but most such studies lack sufficient resolution to determine the number and sizes of regions of low migration or to identify the factors responsible for them . Here we report on high resolution genomic scans of migration/differentiation across several pairs of sunflower ( Helianthus ) taxa that vary in divergence time, strength of reproductive isolation, and degree of geographic separation. We show that genomic regions of differentiation are small, except in areas of low recombination, such as near the breakpoints of chromosomal rearrangements. We also demonstrate that the degree of geographical isolation is a better predictor of patterns of introgression than the strength of biological isolation. On a more general level, our results add to a growing body of evidence indicating that the extent of introgression is far greater (and the effects of 'speciation genes' far smaller) than implied by speciation texts. Thus, the genomes of hybridizing plant species are complex mosaics that exhibit fine-scale variation in ancestry.
Divergent natural selection and plant speciation
Ortiz-Barrientos, D 1
1 The University of Queensland, Brisbane, Australia
Natural selection sometimes drives the evolution of new species as traits favoured within populations may incidentally create reproductive isolation between populations. Here, I synthesize the various mechanisms by which natural selection can lead to the evolution of reproductive isolation, and discuss its impact during the early stages of divergence. I discuss recent data on the origin of Senecio pinnatifolius parapatric ecotypes and the various forms of reproductive isolation emerging between them. I combine ecological and population genomics approaches to demonstrate how both biotic and abiotic selective agents maintain morphological differences in the face of gene flow, and discuss the role of coadapted gene complexes and genetic architecture in the initiation of reproductive isolation between populations. Finally, I show how drift and natural selection interact to produce striking patterns of genomic differentiation across large expanses of land, possibly leading to adaptive radiations and the parallel evolution of traits and species. Altogether, recent results on plant speciation provide strong evidence for the role of natural selection in creating reproductive barriers between plant populations, and suggest that their genetic basis could lead to rapid speciation.
From genes to GIS: the origin of diversity in
Solanum
Moyle, L 1
1 Indiana University, Bloomington, USA
Adaptation and reproductive isolation both contribute to the formation of new species and therefore the generation of biodiversity. Nonetheless, surprisingly little is known about the specific mechanisms that underpin these two fundamental evolutionary processes. Working with tools that range from species distribution modelling to finescale genetic mapping, we are using the plant group
Solanum section Lycopersicon (the wild tomatoes) to dissect the genetics and evolution of adaptive trait differences and reproductive isolating barriers among species. Tomatoes are separated by a range of pre- and postzygotic isolating barriers; our Quantitative Trait
Locus (QTL) mapping indicates that the genetic architecture of these barriers is oligogenic and strongly epistatic. Species also differ markedly in phenotypic, physiological, and biochemical traits; our modeling and quantitative genetic analyses indicate that much of this variation is due to adaptive responses to natural variation in abiotic and biotic environments. Using ecological, evolutionary, and genomic approaches, our goal is to integrate our investigations of both adaptation and isolation, to understand mechanisms of evolutionary diversification in this charismatic and biologically diverse group.
Strigolactones: their roles as endogenous hormones
1 and rhizosphere signals
Yamaguchi, S 1
RIKEN Plant Science Center, Yokohama, Japan
Shoot branching is a major determinant of plant architecture and is highly regulated by endogenous and environmental signals. Previous studies using a group of enhanced shoot branching mutants suggested the involvement of a novel carotenoid-derived hormone in inhibiting the outgrowth of axillary buds. We have recently shown that strigolactones (SL), a group of terpenoid lactones, act as this new hormone or its biosynthetic precursor. SLs were first discovered in root exudates in the 1960s, as seed germination stimulants of root parasitic plants, such as Striga and Orobanche species. Later, they were shown to have a function in symbiotic interaction with arbuscular mycorrhizal fungi, which facilitate the uptake of inorganic nutrients by plants in the rhizosphere. Thus, SLs act as an endogenous hormone as well as being released from roots as an allelochemical in symbiosis and parasitism with neighboring organisms. Earlier studies have demonstrated that SL production is highly elevated by
6

phosphate and/or nitrate starvation in several plant species, which is thought to contribute to successful symbiosis with arbuscular mycorrhizal fungi. Our recent experiments using rice mutants suggested that the elevated SL production under phosphate-deficient conditions also plays a role in minimizing shoot branching for efficient utilization of the limiting phosphorus resource. These findings support the idea that the dual role of SL as an endogenous hormone and a rhizosphere signal might be closely related to adaptive responses to inorganic nutrient availability.
Secreted peptide signals required for maintenance of root stem cell niche in
Arabidopsis
Matsubayashi, Y 1
1 Graduate School of Bio-Agricultural Sciences, Nagoya
University, Japan
Secreted peptides are now recognized as important members of intercellular signals that coordinate and specify cellular functions in plants. Some of the secreted peptide hormones undergo complex post-translational modifications that are mediated by specific enzymes which recognize multiple target peptides. Because such modifications are generally critical for the functions of individual peptide hormones, the presence of novel peptide hormones should be revealed through phenotypic analysis of the mutants of post-translational modification enzymes. Tyrosine sulfation is a post-translational modification that has been found in several peptide hormones in plants. This modification is mediated by tyrosylprotein sulfotransferase (TPST). In loss-offunction mutant of TPST (tpst-1), root meristematic activity is considerably decreased and root stem cells are not maintained. Because known tyrosine sulfated peptide hormones did not recover these defects of tpst-1, we speculated that an as-yet undiscovered tyrosine-sulfated peptide(s) regulates root meristematic activity and maintenance of the stem cell niche in Arabidopsis . To identify this peptide signal, we searched the Arabidopsis genome for genes likely to encode sulfated peptide(s), determined their mature peptide structures and tested their activities to recover defects of tpst-1 mutant. We identified a novel family of functionally redundant sulfated peptides that restore root stem cell and transit amplifying cell proliferation in tpst-1. We named these peptides root meristem growth factors (RGF). RGFs are expressed mainly in the stem cell area and the innermost layer of central columella cells. We also found that RGF1 defines expression levels and patterns of the root stem cell transcription factor PLETHORA mainly at the posttranscriptional level. The RGFs function independently of the auxin pathway. Our results reveal that secreted peptide signals play a crucial role in root meristem development . Reference: Matsuzaki Y. et al. 2010.
The quest for florigen: are we there yet?
Schmid, M 1
1 Max Planck Institute for Developmental Biology,
Tübingen, Germany
The induction of flowering is a central event in the life cycle of plants. When timed correctly, it helps ensure reproductive success, and therefore has adaptive value.
Because of its importance, flowering is under the control of a complex genetic circuitry that integrates environmental and endogenous signals, such as photoperiod, temperature and hormonal status. Based on grafting experiments, it has been long proposed that photoperiod is perceived in leaves where it leads to the induction of a flower-forming substance, or 'florigen'.
The florigen is then thought to be transmitted to the shoot apex where it induces the transition to flowering. The molecular nature of florigen has eluded characterization for 70 years. Genetic analyses in Arabidopsis thaliana have identified mutations in a number of genes such as
CONSTANS (CO) and FLOWERING LOCUS T (FT) that control flowering in response to inductive photoperiod. Whereas accumulation of the CO protein appears to be at the core of measuring photoperiod, recent data suggested that the FLOWERING LOCUS T
(FT) protein constitutes a mobile signal that transduces the information to induced flowering from the leaves to the shoot apex. Interestingly, the function of the CO/FT module seems to be evolutionary conserved, as mutations in homologous genes in other species also disrupt flowering in response to inductive day length. The regulation of FT expression, the relative contribution of
FT mRNA and protein movement to the floral transition and finally the role of FT in orchestrating the floral transition at the shoot apex will be discussed.
Female gametophyte patterning and function in grasses
Dresselhaus, T
ML 1 ,
Sprunck, S 1
1
Amien, S
, Krohn, N
1
1 , Srilunchang, K
, Juranic , M 1
1 , Marton,
, Leljac-Levanic, D 1 ,
1 Cell Biology and Plant Biochemistry, University of
Regensburg, Germany
After three rounds of mitotic nuclei divisions, the female gametophyte of flowering plants (angiosperms) develops from a megaspore into a multicellular haploid structure harbouring two female gametes, egg and central cell, respectively. These gametes produce embryo and endosperm after the angiosperm-specific process of double fertilization has been executed. In addition to gametes, the female gametophyte contains accessory cells, namely two synergid cells representing glandularlike cells involved in pollen tube attraction and perception as well as up to 40 antipodal cells in grasses.
During the past few years, the female gametophyte emerged as an exciting system to study fundamental biological question in plant cell and developmental biology such as nuclei migration and karyogamy, cellularization, establishment and maintenance of cell specification and polarity as well as cell recognition and activation mechanisms. We are studying maize as a model as it generates a typical grass female gametophyte containing large cells allowing their manual dissection.
Using transcript profiling of isolated female gametes and accessory cells, we found a number of genes being specifically up-regulated in either of the female
7

gametophyte cell types. Among these especially transcripts encoding predicted small secreted proteins and those for proteins involved in post-translational protein modification are highly abundant and have been selected for functional studies. We will report here about the role of a MATH-BTB protein homologous to animal
MEL-26 being known for its role in cytokinesis and as a component of an E3-ligase for meiosis-mitosis progression. A gene encoding a grass-specific diSUMOlike protein was identified as a regulator of nuclei positioning and cell viability of the female gametophyte.
Other genes for small secreted proteins being involved in cell specification include an EA1-box protein or small cysteine-rich proteins (CRPs), which play major roles for pollen tube guidance, burst and sperm delivery. Although plant hormone biosynthesis genes seem to be lowly expressed inside the immature female gametophyte, exogenous hormones, especially auxin, seem to play an important role to polarize the female gametophyte also in grasses .
Recognition of 'self': conversations that commit 'self' pollen to suicide
Franklin-Tong, N (VE) 1
1 University of Birmingham, UK
Cellular responses rely on signalling. Selfincompatibility (SI) is an important geneticallycontrolled mechanism used by many angiosperms to prevent self-fertilization and inbreeding. A multi-allelic S locus allows discrimination between ‘self’ (incompatible) pollen from ‘non-self’ (compatible) pollen when it lands on the stigma. Interaction of matching S-determinants from the pistil and pollen to allow ‘self’ recognition, which triggers rejection of incompatible pollen. In
Papaver rhoeas (poppy) the S-determinants are PrsS
( Papaver rhoeas stigma S) and PrpS ( Papaver rhoeas pollen S). PrsS is a small novel cysteine-rich protein that acts as a signalling ligand that interacts with its cognate pollen S-determinant PrpS, and represents one of the few examples of plant ligands whose interactors are known.
PrpS is a small novel transmembrane protein, so is interestingly, not a ‘classic’ receptor. Interaction of PrsS with incompatible pollen stimulates SI-specific Ca 2+ influx. We recently identified SI-activated conductance(s) that is not voltage activated, but is voltage sensitive. Evidence suggests that it may represent a Non-Specific Cation Channel (NSCC). Recent data also implicate ROS and NO signalling. Downstream targets include the actin and microtubule cytoskeletons, a soluble inorganic pyrophosphatase, Pr-p26.1, and activation of a MAP kinase, PrMPK9-1. Ultimately, and probably the major target for SI signals is initiation of programmed cell death (PCD) involving several caspaselike activities in incompatible pollen. This provides a very neat way to get rid of unwanted ‘self’ pollen and prevent self-fertilization. I will describe recent progress in identifying the signaling components and downstream targets of SI signals, and will attempt to provide an overview integrating our knowledge of how this SI system operates. We have recently transformed
Arabidopsis thaliana , which is a self-compatible, with
PrpS, and I will present data showing that when transgenic pollen expressing PrpS interacts with recombinant PrsS protein, it elicits a remarkably similar
8 response to that triggered in incompatible Papaver pollen. This demonstrates that Papaver PrpS is functional when transferred to an unrelated species and suggests that Arabidopsis has all the cellular machinery required to elicit the downstream events triggered by interaction of PrsS and PrpS. These preliminary data suggest that the poppy SI system may be potentially useful for manipulating crop plants to produce F1 hybrids more easily.
Pollen allergens – molecules with impact beyond plant reproduction
Singh, MB 1
1 ARC Centre of Excellence for Integrative Legume
Research, Melbourne School of Land and Environment,
The University of Melbourne, Parkville, Australia
Pollen allergy is the most common elicitor of seasonal respiratory allergic diseases such as hay fever and seasonal asthma. The prevalence of both hay fever and allergic asthma has increased significantly in recent decades. Recent research has pointed towards the possible stimulatory effects of global environmental change on pollen allergen production and thus the potential for pollen-related allergic symptoms to increase. These allergic symptoms that are based on immunoglobulin E (IgE) recognition of specific allergen molecules affect almost 20% of the population worldwide. After landing on the moist human respiratory epithelial surfaces, individual pollen grains release copious amounts of soluble proteins that are potential triggers of immediate immunological hypersensitivity symptoms. This presentation will focus on grass pollen allergens; their structural properties, biological roles and allergic potential, and current efforts to generate vaccines for immunotherapy of pollen induced allergic diseases.
Improving projections of plant species and ecosystem
1 responses to climate and atmospheric change
Midgley, G 1
SANBI, South Africa
Current projections of species and ecosystem responses to anticipated climate change have well known limitations, but advances are being made that will improve our understanding of the risk of these changes to plant biodiversity. In this paper I will outline how greater integration and coordination between modeling, experimentation and monitoring efforts are required to develop our ability to make more credible projections of species and ecosystem shifts as climate and atmosphere continue to change. It is particularly important to recognize that the environmental changes to which we are committed over the next decade or two provide an opportunity to test our predictive capabilities, but only if we are able to coordinate efforts to make the required observations.

Environmental change and the rise of flowering plants in evolutionary time
Bond, W 1
1 Botany Dept, University of Cape Town, South Africa
As we face a period of rapid projected climate and environmental change, it is useful to explore how the world’s modern flora came to be over evolutionary time scales. Flowering plants are now the dominant component of the world’s flora, but the ways in which angiosperms achieved this dominance are still being elucidated. Adaptive responses to key environmental changes, including climate, fire regime and atmospheric
CO
2
concentration, seem all to have been critical in the rise of the angiosperms. This deep time perspective provides an important context for appreciating the magnitude of ongoing and projected anthropogenic changes to the world’s disturbance regimes, climate and atmosphere, and for understanding the adaptive potential of modern floras in the face of these changes.
Climate change susceptibility and adaptability: how much is too much change for global species?
Foden, W 1
1 IUCN, Cambridge, UK
There is growing evidence that climate change will become one of the major drivers of shifts in species dominance and geographic range, and even species extinctions, in the 21st century. How might we begin to predict which species will be more vulnerable to climate change effects than others? The prediction of these relative vulnerabilities has most commonly been carried out using models of geographic range change, which have many well-known limitations. I present a collaborative IUCN-led study that introduces an alternative approach based on species' traits, which provides an opportunity to take into account species biology (e.g. demographic, physiological and ecological traits) and expert knowledge that may be derived from species' evolutionary history (e.g. fire damage susceptibility and ecological plasticity). Based on assessments of individual species' biological sensitivity, their predicted climate change exposure (derived from
General Circulation Model projections), as well as their anticipated adaptability to climatic change, the study has assessed relative climate change susceptibility of global birds, amphibians and corals, as well as various plant groups from Southern and East Africa. I will highlight some of the species groups predicted to be at greatest risk from climate change and identify the regions of their greatest concentrations. Exploring the relationship between pilot species' predicted climate change vulnerability and existing Red List status allows an assessment of future threats in terms of this measure.
These results present a new kind of assessment for climate change impacts on species dominance shifts and potential extinctions. I will discuss implications of the findings for global plant conservation, including on the development of strategies for assisted colonisation and management of possibly novel communities of the future.
Megatrends and ecological intensification of major
1 cropping systems
Cassman, K 1
University of Nebraska, Lincoln, USA
Achieving climax human population of about 9.5 billion and then an orderly decline can occur this century if economic development can alleviate poverty and support universal education. Adequate food supply at reasonable cost is a prerequisite because high food costs reduce economic growth rates and undermine political stability.
Unfortunately, yield growth rates of our major food crops are not sufficient to meet expected food demand without massive expansion of crop production at expense of remaining carbon-rich and biodiverse rain forests, wetlands, and savannah. Avoiding this scenario requires accelerating yield growth rates while concomitantly reducing the environmental footprint of agriculture through a process of ecological intensification. Regions will suitable soils and climate to support high yields must produce near the yield potential ceiling (Yp)determinedby biophysical limits set by temperature, water supply, and solar radiation. In some cropproducing regions, however, yields are already plateauing because average farm yields are 80-85% of
Yp. In other regions, current cropping systems are degrading soil and water quality because climate and soils cannot support intensive agriculture. A global atlas that estimates the difference between yields and Yp on all currently farmed land can identify where crop yields are likely to plateau and where large exploitable yield gaps justify investments in ecological intensification.
Ensuring food security also requires continued breakthroughs in crop genetic improvementand in management of soil, water, and nutrients to increase resource capture and the efficiency with which resources are used.
Integrated approaches to accelerate crop genetic improvement
Richards, R 1
1 CSIRO, Canberra, Australia
The rate of genetic improvement in our major crops threateningly lags behind the anticipated future food requirements of our growing global population. We must accelerate the rate of grain yield improvement if we are to avoid future food catastrophes. Where will genetic increases come from? Empirical plant breeding will remain the cornerstone of genetic improvement as this effectively integrates yield improvement with tolerance/resistances to pests and diseases and with grain marketability. However, alone this is not going to be enough. Genetic engineering in the next 20 years will make major contributions to pest and disease resistance and will result in improved management practices but it is unlikely to result in significant genetic advances in yield potential or tolerance to abiotic stress. The main advances in the next 20 years are likely to be achieved
9

from (i) smarter, faster and cheaper phenotyping and genotyping methods and (ii) identifying key traits responsible for the improved capture of the resources required for crop growth (light, water, nutrients) and then convert this to grain. These traits will be required to design and breed crops for target environments and for management practices. A challenge is that the most important traits contributing to increased yield under both favourable and unfavourable conditions are multigenic and can be difficult to select in breeding programs. Beyond 20 years is difficult to predict .
Hopefully photosynthesis will then be more amenable to improvement than it is now.
Can we increase net primary productivity of major food crops?
Amthor, J 1
1 University of Sydney, Australia
In the face of our growing population, finite arable land area, and desire for globally increased standards of living, the need for further crop yield increase is inevitable. Enhanced crop net primary production (NPP) is one mechanism, among others, to increase yield.
Extending crop growth duration could increase NPP and so too might 'improvements' to primary metabolism– specifically, faster photosynthesis, slower photorespiration, and/or more efficient growth and respiration. These improvements to primary metabolism are considered herein. In principle, faster photosynthesis might be achieved by improved kinetics of photosynthetic enzymes, particularly that of rubisco.
Some possibilities are reviewed, but past successes are limited. Conversely, the demonstrated ability to shortcircuit the photorespiratory pathway in intact plants, and to release the resulting CO
2
directly within the chloroplast, could be the foundation for significantly increased crop NPP. Also, revived interest in converting rice from a C
3
to a C
4
crop (with attendant increase in
NPP) is intriguing, even if yet unproven. Slowing crop respiration might increase NPP, but caution is warranted because rapid growth requires rapid respiration and some
'maintenance respiration' is essential. A knowledge gap remains concerning the possibility of otiose respiration, and how it would be eliminated if found. Speculation about time frame for improving primary metabolism on a farmer's field is just that –speculation. Some advances perhaps might be seen within 10 years, but I expect several decades will pass before significantly improved crop-plant primary metabolism becomes an on-farm reality.
BiodiversityWorld
and the
Catalogue of Life
: tools for studying the ecology and behaviour of the entire global biota in a virtual laboratory
Bisby, FA 1
1 University of Reading, UK
10
The dream of the biodiversity informatics community is to provide a virtual laboratory equipped with electronic tools and datasets with which to study the global biota.
BiodiversityWorld and the Catalogue of Life provide pointers to two themes that will come together with powerful outcome. Significant progress with the
Catalogue of Life , alongside partners including ITIS,
WoRMS, FADA, and The Plant List , means that we are close to having an electronic catalogue of the world's known species, including Plants, Animals, Fungi and
Micro-organisms. This taxonomic backbone provides a framework for species-based global datasets. Initiatives to assemble ecological and functional traits across taxa on a global scale are now feasible, as are the possibilities for Taxa of the World and Biota of the World services for access to comparable species lists for each area of the world and potentially for each biome and ecosystem.
Converging with this growth of datasets is the idea behind the BiodiversityWorld pilot-project. A suitably empowered distributed computing environment could provide the platform, both for access to global biodiversity datasets, and for libraries of analytical routines, a toolbox of agreed and experimental workflows, and workspace for wide-ranging experimental, analytical and modelling investigations: a true virtual laboratory or cyber-infrastructure. This convergence of platform and data sets will enable the start of the interesting part: the elucidation of models, processes and even rules for the behaviour of biodiversity at the large scale – a space we need to occupy, and the start of a truly evidence-based global biodiversity science.
Intelligence systems for emerging plant pests and
1 diseases
Burgman, M 1 , Nunn, M 1 , Lyon, A 1
The University of Melbourne, Melbourne, Australia
Plant pests and diseases are dealt with most efficiently when they are identified early, and ideally, before they arrive in a susceptible area. Biosecurity intelligence encompasses a range of methods for gathering, filtering and communicating information on emerging threats, so that decisions can be made early to mitigate risks. This presentation outlines three strategies for biosecurity intelligence: web-based intelligence software, informal social networks, and foresighting. Web-based systems for animal diseases are relatively well developed. Our research shows they have a range of features that may be useful for plant biosecurity. Foresight activities are extremely useful for assisting users to tune filters to identify critical pieces of information. Professional networks may be supported and developed to disseminate biosecurity information so that it contributes effectively to decision making .
A risk assessment protocol for managed relocation
Possingham, H 1 ,
Martin, T 1
Rout, T
, Armstrong, D 1
1 , McDonald-Madden, E 1 ,
1 The University of Queensland, Brisbane, Australia
Managed relocation (sometimes called assisted colonisation) is the deliberate movement of a species to a

location outside its known range in response to unabatable threats . The threat might be climate change, but it could equally well be any other human-induced change that society is unwilling to halt (e.g. land-clearing) . We present a quantitative framework for assessing the relative costs and benefits of such an action that accounts for factors like: the chance the translocation is successful; the chance, and amount, of negative impact; and the negative effect on the source population . If this benefit is positive we show how that, using information about costs, such moves can be prioritised in a rational fashion .
Finally I note, that, despite the resistance to managed relocation, we seem to have done it an awful lot.
Developing durable resistance to potato late blight disease using an understanding of how the pathogen infects its host
Birch, PRJ 1
1 Division of Plant Sciences, University of Dundee (at
JHI), Invergowrie, Dundee, UK
The oomycete Phytophthora infestans is the cause of late blight, the most significant global disease of potato.
Notoriously, it precipitated the Irish Potato Famine in the mid-19 th century – seldom can a plant disease have had such a dramatic effect on the social, political and demographic structure of a country. More than a century of breeding for late blight resistant potatoes has met little success, as most deployed resistances are overcome in a few growing seasons. Late blight control has thus relied heavily on application of prophylactic chemicals. In
Europe, new legislation is set to reduce the range and levels of chemical application, driving an urgent need to better understand the molecular basis of late blight disease, in order to seek new approaches to combat infection.
Over recent years we have learned that, during infection,
P. infestans delivers proteins called RXLR effectors inside living potato cells. One clear role of these effectors is to suppress the plant immune system, creating an environment in which the pathogen can colonize its host.
The genome sequence of P. infestans has revealed hundreds of candidate RXLR effector genes, residing in regions rich in transposons where they are likely to evolve rapidly. We have learned also that these effectors are the ‘targets’ of resistance receptor genes that are also evolving rapidly in natural populations of wild potato species. I will describe how we have been studying
RXLR effectors, finding those that are critical to late blight disease and are prevalent across the pathogen population. Such effectors have sparked a search, in wild potato collections, for resistance genes that recognise them. I will argue that the careful combination of such resistances in potato cultivars will provide late blight disease resistance that is more durable.
Reducing the threat of wheat rusts to global food security
Park, RF 1
1 The University of Sydney, Plant Breeding Institute,
Narellan, Australia
Wheat is the most important cereal crop, and the three rust diseases that infect it are among the most important biotic impediments to stable wheat production in many regions. The development of our understanding of the wheat rust pathogens, their hosts, and the interactions between these two organisms, is a remarkable chapter in the history of the plant sciences. The work began in earnest in the latter part of the 1880s in Australia with a series of Rust-in-Wheat conferences, which followed a stem rust epidemic in 1889 that was estimated to have caused some £2–3 million in losses. While advances in our understanding of this host : pathogen system have permitted considerable success in developing and deploying rust resistant wheat cultivars, these diseases continue to impact on global wheat production principally because of their abilities to overcome the resistance genes incorporated by breeders. The two most significant wheat rust challenges to emerge over the past
10 years have been the intercontinental spread of two pathotypes of the stripe rust pathogen with increased aggressiveness in the early 2000s, and the detection of stem rust pathotype 'Ug99' in 1999 Uganda and its subsequent spread to Kenya, Ethiopia, Yemen, Iran and
South Africa. In both cases, regional epidemics have reduced production and increased costs to growers due to the need for fungicide intervention. These challenges have refocused international rust control efforts, providing a strong impetus to develop and implement new tools to monitor rust pathogens and accelerate the development and adoption of rust resistant wheat cultivars.
The genome of the wheat and barley crown rot pathogen
Fusarium pseudograminearum
reveals horizontally transferred virulence functions
Gardiner, DM 1
M 3 , Kazan, K 1 , Chakraborty , S 1
1
Manners, JM 1
, Solomon, P 2 , McDonald, M
,
4 , Marshall,
McDonald, B 4 ,
CSIRO Plant Industry, Queensland Biosciences
Precinct, Brisbane, Australia; 2 Research School of
3
Biology, College of Medicine, Biology and Environment,
The Australian National University, Canberra, Australia;
Queensland Facility for Advanced Bioinformatics, The
University of Queensland, Brisbane, Australia; 4 Plant
Pathology, Institute of Integrative Biology, Zürich,
Switzerland
Fusarium crown rot disease is a chronic problem in wheat and barley in arid environments such as Australia and there are currently no fully resistant wheat cultivars.
In Australia, F. pseudograminearum is the pathogen predominantly associated with crown rot although related
Fusarium species such as F. graminearum and F. culmorum have the ability to cause crown rot in some regions. These three Fusarium species can also cause globally important head blight disease of wheat. To increase our understanding of factors affecting pathogen
11

virulence, we have sequenced the genome of an F. pseudograminearum isolate and compared it to the publically available genome sequence of F. graminearum . Despite overall sequence conservations, striking differences have also been observed between the genomes of these two Fusaria. Most strikingly we also identified a gene encoding an amidohydrolase that appears to have been acquired by horizontal gene transfer. This gene has a clear orthologue in the genome of the wheat pathogen Stagonospora nodorum but not in any other fungal genome and the next closest sequence matches are from bacteria. Deletion of this gene from F. pseudograminearum resulted in a reduction in virulence on barley but not wheat. Population studies suggest this acquisition of this gene in both species may have been independent. Its presence in these two unrelated pathogens but not in any other fungal species suggests a role for this gene in a common pathogenesis mechanism that targets an important defence pathway in cereals .
Coalescent global communities for species-level plant taxonomy
Knapp, S 1 , Baker, W 2
1 Dept of Botany, The Natural History Museum, London ,
UK;
UK
2 Royal Botanic Gardens Kew, Richmond, Surrey,
Plant taxonomists have in recent years embraced a number of new initiatives and technologies in attempts to meet global targets and accelerate species discovery and delimitation. It is often assumed that the community of taxonomists describing species is small and fragmented, but recent work suggests this is not the entire picture.
Despite this, the imperative to both increase and accelerate our acquisition of knowledge about the species of plants is great – much attention has been focused on the so-called 'taxonomic impediment'; taxonomic capacity and research itself not keeping up with demand for taxonomic information. Many solutions to this problem have been proposed, ranging from the purely technological (for example, bar-coding and automatic identification) to largely people-centred (for example,
EU and UK-funded projects using the web and NSF's
PBI Initiative). Taxonomy, like physics or astronomy, is most definitely big science and can learn a lot from these other disciplines. Can taxonomists come together as coalescent communities to speak with one voice while retaining the differences and debate characteristic of a vibrant scientific discipline? Does the development of national strategies make sense in a globalised scientific enterprise? Some of the social issues associated with working in large distributed groups of researchers have been explored by a variety of projects over the last few years; the human factors involved in the science of plant taxonomy - both in terms of research partnerships and audiences and how individuals function within institutions are critical to achieving our goals. These issues relate to the use of new technologies as well as to interactions between people themselves. People are often perceived as the problem, but they must be the solution as well if plant taxonomy is to truly take advantage of the
21st century.
Beyond the Plant List: plant systematics in the 21st century
Raven, PH 1 , Hopper, S 2
1 Missouri Botanical Garden, St Louis, USA;
Botanic Gardens Kew, UK
2 Royal
Gaining an adequate knowledge of the approximately
380,000 valid named and described species of plants and finding the perhaps 75,000 or more awaiting recognition poses a significant challenge for the science of plant systematics. We know very little about the great majority of even the named species, many of which are known from only one or a few specimens. Of great use will be
The Plant List, an integrated synonymized list of vascular plants that was produced by the common efforts of the
Royal Botanic Gardens, Kew, and the Missouri Botanical
Garden and put on line at the end of 2010. Continually upgraded over the years, and with many other institutions and individuals – virtually the entire plant taxonomic community – involved, TPL, by indicating directly such details of authorship and bibliographical citation and providing a standard reference to what has already been established, will relieve botanists of much of the routine that currently occupies a great deal of their time.In addition, all of the literature of systematic botany, rapidly being accumulated in the Biodiversity Heritage Library and Botanicus, will soon be instantly available for consultation, and images of type and other specimens will be fully available on line soon. With this substantial mass of information available, systematists should be able to devote themselves to the study of plants in nature, the determination of their characteristics, their role in ecosystems, and their conservation status, using molecular tools, automated character recognition and identification software, rapid imaging, space technology, and many other novel means of understanding them.
Because of the rapid destruction of habitats; spread of alien invasives, pest, and diseases; and especially global climate change, conservation should become the primary object of our efforts in this century, with as many as half of all species at risk of extinction over the coming decades.
Plant DNA barcoding: the future of taxonomy,
1 ecology, and species discovery
Kress, J 1
Dept of Botany, United States National Herbarium,
National Museum of Natural History, Smithsonian
Institution, Washington, DC, USA
How many plant species remain to be discovered and described? As taxonomists, how and where will we find them? As ecologists, how will we investigate their life histories and community interactions? As conservationists, how will we protect and preserve them?
Natural habitats continue to be degraded around the world and the deadline for finding and describing biodiversity approaches at an ever-increasing rate . Now more than in the past we need new scientific tools to supplement our time-tested methods for species
12

discovery and understanding. DNA barcodes provide one such tool . Based on short stretches of DNA that are easily sequenced and variable enough to distinguish among species, barcodes provide a rapid assessment of taxonomic similarity and genetic distinctness . As a first pass in identifying new taxa, DNA barcodes alert us to the existence of unknown species and may lead us to test the validity of previously described taxa . In addition the assembly of DNA barcode libraries for species-rich natural communities greatly enables ecological studies where accurate species identifications are required .
Finally, well-resolved molecular phylogenies of naturally co-occurring species derived from DNA barcode sequences have the potential to improve investigations of the mechanisms that underlie the assembly of plant communities and the evolution of plant functional traits .
DNA barcodes, if applied appropriately, will enhance future research focused on the interface between species discovery, taxonomy, ecology, and evolution .
Patterns in the history of plant diversity: testing, reconciliation and accounting for missing data
Crane, PR 1
1 Yale School of Forestry and Environmental Studies,
New Haven, USA
Evidence of the structure, biology and diversity of extinct plants derived from paleobotanical research over the past
200 years has greatly expanded our view of the history of plant life and provided insights into key events in plant evolution that would have been inaccessible from studies of living plants alone. In general, there is remarkable consistency between the large-scale patterns of plant evolution inferred from studies of living plants and those obtained from studies of fossils . However, two areas have emerged since the last botanical congress where this reassuring consistency seems to have broken down, and where paleobotanical and neobotanical evidence appear to conflict significantly: the pattern of phylogenetic relationships among seed plants, and the antiquity of certain lineages of angiosperms (with the associated implications for the antiquity of certain kinds of biomes) .
In both instances questions of missing data seem to be at the heart of the problem . As a paleobotanist I worry about the impact of extinct taxa on inferences of seed plant relationships based on living plants alone . While acknowledging the inadequacies of current phylogenetic analyses based on structural data I am unwilling to abandon those results entirely in favor of other patterns of relationship that while based on vast amount of molecular data are likely to be from an inadequate sample of taxa . At the same time, those who use molecular dating techniques to determine the age of clades acknowledge the limitations of current models of molecular evolution from which such ages are derived.
Nevertheless, they seem more comfortable with their own results than trusting to the completeness of the fossil record.
In this talk I will argue that neither neobotanists nor paleobotanists will ever have the data that they would ideally want to address these questions, but combined efforts and opportunities for partial reconciliation offer the promise of more robust hypotheses . Fresh perspectives on key questions of plant evolution are possible, but are less likely to come from approaches that fail to account for all of the available data.
The future of plant diversity
Coates, D 1
1 Dept of Environment and Conservation, Western
Australia, Australia
A recent assessment by the Sampled Red List Index indicates that as many as one fifth of the world’s estimated 380,000 plant species are under threat. This suggests that some 76,000 plant species are in danger of extinction from a broad range of threats including habitat loss, habitat transformation, over-exploitation, alien invasive species, pollution and rapid environmental change. Although the disappearance of such a critical component of biodiversity poses one of the greatest challenges for the future it is encouraging that there are now key strategies being developed and implemented which are directed towards addressing and recovering this potential loss. For example, the Global Strategy for
Plant Conservation set 16 outcome-oriented global targets for 2010 that provided an important starting point for improving world plant diversity conservation. One such target Target Eight of The Global Strategy for Plant
Conservation recommends that ‘60% of threatened plant species’ should be ‘in accessible ex situ collections, preferably in the country of origin and 10% of them’ should be ‘included in recovery and restoration programs’ by 2010. Based on current estimates by the end of 2010 some 4,560 plant species worldwide should be in reintroduction programs. Such goals may be ambitious and considered unrealistic by some. However, as shown in Western Australia, with a flora of 12,200 species, they can be achieved and even exceeded with
70% of the threatened flora in ex situ conservation and
13% used in reintroductions. Equally the goals for ecological restoration can set very high benchmarks but a bold and proactive approach where actions are implemented now, perhaps without full knowledge, but in an adaptive management framework will need to considered if successful plant diversity conservation is to be achieved. Loss of some plant diversity is inevitable but the magnitude of that loss will depend on our ability to implement strategies already in place, develop new strategies and integrate current knowledge in plant conservation science with on ground actions. For example, the introduction of plant species outside their natural range in response to the threat of climate change has been raised recently as a key tool in plant translocations. While achieving long term persistence particularly in the face of rapid environmental change may require a reassessment of how we manage genetic diversity, maximise evolutionary potential and deal with local adaptation. Also recent strategic approaches to combining ex situ and in situ conservation that involve the creation of artificial populations of plants that can be subsequently used for reintroductions and restoration have considerable potential for enhancing plant diversity conservation. These approaches will be discussed in the context of future goals and actions that will be needed to halt and even reverse the global loss of plant diversity.
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Plant diversity today: how well do we know what we have now?
Nic Lughadha, E 1 , Allkin, R 1 , Rivers, M 2 , Saltmarsh, A 1
1
2
Royal Botanic Gardens, Kew, Richmond, UK;
University of St Andrews, St Andrews, Scotland
The volume of plant diversity data available on our desktops has increased by orders of magnitude over recent years. New technologies and reductions in the costs of digital storage have greatly facilitated the capture of new information and the conversion of existing information to digital form. Most of us now have at our fingertips more data on plant diversity than we could hope to explore in lifetime . But how effective have our efforts been to analyse, synthesize and apply this information? How good an overview of plant diversity do we have currently? Estimates of the number of known plant species have varied widely, though the creation of consolidated checklists of accepted species with synonymy has reduced the range of likely values. Full species lists have been published for several megadiverse countries but most still lack a basic inventory of what is known . Species considered new to science continue to be described at a steady rate but inventory effort is taxonomically and geographically patchy. Current data provide limited support for reliable estimates of how many species remain to be discovered . Distribution data at country level or below are readily available for well over half of all known species, but most of the point data for most species remains locked up in our herbaria . Thus we can identify hotspots of diversity and endemism on a broad scale but generally lack the data required for detailed analysis and conservation planning more locally.
The vast majority of online records relate to widespread, temperate plant species and though we can find at least one image for most plant species these images are often far from ideal for identification purposes . As systematic botanists we recognize that any failure to synthesize and to present the results of such synthesis more effectively has consequences not only for related disciplines such as conservation and ecology but also, more broadly for human health and welfare. . In this review we attempt a score-card for our knowledge of plant diversity in 2011 and consider how targeted efforts over the next decade might address deficiencies in our current overview and our ability to organize current knowledge .
Making a botanical institute (nearly) from scratch
Sohmer, S 1
1 Botanical Research Institute of Texas, Fort Worth, USA
The Botanical Research Institute of Texas (BRIT) was formed on paper in 1987. Based on the collections accumulated at Southern Methodist University, BRIT moved into warehouse space in downtown Fort Worth in
1991, has grown its herbarium from about 400,000 specimens to the nearly 1,100,000 today, and its
14 horticultural and botanical library from about 50,000 books and journals to about 125,000 during the same period. The budget has grown from about $175,000 per annum in 1993 to nearly $3,000,000 today, and its staff from three to over 30. All of its operating costs are obtained as donations from the private sector and most of its projects are funded by grants from private foundations and state and government agencies, such as the National
Science Foundation. BRIT has established leading programs in horticulture, taxonomy, ecology and database management in the Peruvian Amazon, major collection activities in Papua New Guinea and the
Philippines, and is producing the definitive volumes on the flora of Texas through the Illustrated Flora of Texas
Project. It maintains a leading press publishing a respected scientific journal and a number of significant books each year. It has nearly completed a $48 million capital campaign, the first in its history, that has resulted in a spectacular facility that will be only the 8 th LEED platinum level building in the entire state of Texas. There are 168 geothermal wells, solar panels on top of the collection wing, a piece of the Fort Worth Prairie growing on top of the research wing, and all manner of sustainable and material in the building. Dr Sohmer will focus on the means by which this institute arose from nearly nothing but a dream and has thus far prospered almost entirely on private philanthropy.
Sing of Botany
Silk, WK 1, 2
1 Dept of Land, Air, and Water Resources, University of
California at Davis, Davis, CA, USA
One in five plant species – the basis of life on earth – are estimated to be under threat of extinction; sickness of pollinators is threatening food production; and pollution is threatening human health . More than ever in human history there is need for new access to scientific knowledge and new paradigms for collaboration. At the
University of California at Davis, biologist Diane Ullman has established an ArtScience fusion program in the hope that artists will better access scientific literacy, while scientists will better access art as a means of expression .
In 2009 the course 'Earth, Water, Science, Song' was introduced to the curriculum. Students write and perform songs to communicate their understanding of the lectures and readings in environmental science. Instruction features conventional science lectures, participation of guest artists from the community, and studio sessions on music theory, lyric writing, and performance skills. The hope is that the act of translating from science lectures to an art form will foster understanding of the science.
Assigned topics for songwriting include the hydrologic cycle, the carbon cycle, the nitrogen cycle, spatial and temporal variation, soil formation, eutrophication, and fundamental processes such as photosynthesis and transpiration . Instead of writing a final exam the students perform in a university lecture hall and in a local art gallery. A spirit of intense collaboration developed with the collaborative work among musicians and music neophytes, science majors and nonmajors. The repetition of rehearsals fostered retention of course content .
Student energy was high. Students became teachers, as their performances attracted a large audience from the

community . In this lecture-demonstration we will see some videos of botanical songs and hear songs designed to inform, motivate, and celebrate research progress in botany.
2 Work supported by the US National Science Foundation under Grant No. RCN-UBE 0956196. Any opinions, findings, and conclusions or recommendations expressed in this material are those of the author and do not necessarily reflect the views of the National Science
Foundation.
PU01: Fruits of the vine – future climates and wine
Barlow, S 1
1 The University of Melbourne, Melbourne, Australia
Wine produced from the grapevine, Vitis vinifera , has been consumed for more than 10,000 years, evolving alongside cereals as essential part of early civilizations in the Middle East. Since those times many of our great civilizations have evolved and developed the sophistication of both viticulture and winemaking to provide wines of distinction and consistency. A key part of this development has been the matching of particular wine varieties, such as Shiraz and Chardonnay ,to special combinations of soils and climate allowing them to fully express their characteristic flavor and aroma profiles.
This French concept of terroir encompasses the combination of climate, soils, geology and topography, that determines the characteristics of the grapes and ultimately the wines. Wine regions have effectively
'patented' and trademarked the successful combinations of terroirs and grape varieties that produce great wines such as Bordeaux and Burgundy in the form of French appellation system. Climate change challenges these established terroir relationships by changing the climate component thereby altering ripening temperatures and the resulting wines. Rising temperatures in wine regions around the world are resulting in earlier ripening and earlier vintages. In some areas of Australia and Europe vintages have moved forward by as much as a day per year over the past 30–40 years. While some of these vintage temperatures are within the range of season to season variation many are not and certainly will not be in the future. How has climate change altered the climate in well known terroirs and how will the global wine industry respond to these challenges to established terroirs particularly if we ,as consumers, wish to continue to savior the 'grassiness' of Malborough Sauvignon Blanc or the 'white pepper' of cool climate Shiraz ? What sort of wines can we look forward to in the future ?
PU02: The world of plants
Raven, PH 1
1 Missouri Botanical Garden, St Louis, USA
We depend for all fundamental aspects of our lives on plants. The dominant angiosperms evolved at least 120 million years ago, diversified in the face of climate evolution and change, coevolved with insects and other herbivores, and adapted to diverse habitats. Angiosperms provide our food, most of our medicines, building materials, cloth, chemical feedstocks, and many other products and possibilities for the future. In addition they provide an enormous array of ecosystem services and add meaning and beauty to our lives. At least 400,000 species of angiosperms exist, the great majority of them poorly known, with perhaps 20% of the species and a much higher proportion of the genetic diversity threatened with extinction over the next decade or two and probably more than half by the end of the century. In the face of human pressures on the environment, already estimated to be using a rapidly growing 150% of the world’s sustainable productivity on an ongoing basis, we must find effective ways of conserving the plants on which we depend and which offer so much promise for the future.
Learning about them and disseminating the information efficiently, conserving natural areas in the face of growing adverse changes, building seed banks, and educating people to know and love what they are losing – these are elements of the strategy now so badly needed to ensure a future that begins to match the present.
PU03: Brave New World: can we solve tomorrow’s environmental and energy problems by using life itself? – Public discussion panel 1
1 Supported by Royal Botanic Gardens and Domain
Trust, Sydney, Australia; coordinated by Janelle
Hatherly, Manager Public Programs.
Studying life in all its forms is exciting at this time of great technological change. Computers and modern scientific techniques have provided us with an understanding of life processes at the molecular level in a way never before possible. Yet we know little about the unicellular organisms that make up most of the Tree of
Life. Much of our scientific research efforts and investments go into the study and conservation of relatively few multicellular creatures and ecosystems.
Research on the rest of life focuses mostly on controlling harmful microorganisms rather than looking for useful ones.
Is the time right to prioritise research into useful microbes, harnessing them to convert significant amounts of CO
2
into biomass and biofuels and to capture and store significant amounts of carbon to slow climate change? Plants feed us and nature sustains us, but could microorganisms give us the ‘biggest bang for our buck’?
These questions will be discussed and debated by a distinguished panel, moderated by Robyn Williams, eminent Australian Broadcasting Commission science journalist. The format is conversational with Robyn setting the scene and then each speaker presenting a point
15

of view for 5–10 minutes. Then Robyn will ask/take questions – from the floor or via live links.
Speaking for the plants:
* Prof. David Mabberley, Royal Botanic Gardens, Kew,
UK; soon to be Executive Director of Sydney’s Royal
Botanic Gardens and Domain Trust
* Dr Kevin Thiele, Curator Western Australia
Herbarium, Perth, Australia
Speaking for the microbes:
*Dr Jeff Powell, Microbial ecologist and lecturer,
University of Western Sydney, Australia
*Assoc. Prof. Kirsten Heimann, Cell biologist and biofuels expert, James Cook University, Townsville,
Australia
Sister Water Lily meets the Big Bad Banksia Man
Bernhardt, P 1 , Meier, R 1
1 Saint Louis University, St Louis, USA
Can a whimsical and largely discarded branch of illustration be used to reinvigorate botanical education?
We review the works of C.M. Barker (England), W.
Crane (England), May Gibbs (Australia), J.J. Grandville
(France) and M.T. Ross (America). All produced detailed illustrations featuring anthropomorphic flowers, stems and edible plants. The tragic J.I.I. Gerad (a.k.a.
Grandville, 1803–1847) began this trend in floral fantasy to amuse a mature audience of sophisticated Parisians but his techniques were assimilated by later author/artists of children’s books. Within little more than a century
(1847–1952) their combined oeuvre reminded or taught viewers simple, amusing and often visually accurate lessons in plant morphology/identification, phenology, economic botany, plant-animal mutualisms and phytogeography. These drawings, often reinforced botanical information by employing visual puns and satirizing the old, European, ‘language of flowers’. We could certainly use a little of this style today with the fundamentals of plant biology taught so infrequently in many countries. In particular, author/illustrator, May
Gibbs (1879–1969) developed the art of ‘people plants’ to introduce generations of Australian children to plant diversity common to southeastern and southwestern
Australia.
The
Atlas of Living Australia
: infrastructure for
1 biodiversity research
Hobern, D 1
Atlas of Living Australia, CSIRO Entomology,
Canberra, Australia
The Atlas of Living Australia is a national initiative focused on making Australia's biodiversity information more accessible and useable online. In short, 'an online encyclopedia of all living things in Australia' . The Atlas website already holds more than 23 million distribution records for Australia's fauna and flora, integrated with over 300 environmental layers for mapping and analysis.
In addition the site brings together photos, maps, identification tools, reference lists of species names and classifications, databases on biological collections and literature. All of these resources are freely available at http://www.ala.org.au/ . Ultimately, the Atlas aims to enable anyone to find, analyse and map information on all aspects of Australian biodiversity online . Researchers, managers and others can use the Atlas in their efforts to protect, manage and monitor Australia's biodiversity and biosecurity, from conserving species to tracking invasive pests . As a research tool, the Atlas can help to create species distribution models, predict areas that could be suitable for a species, or work out how a species will be affected by a change in climate, among other things . The
Atlas enables researchers to provide policy and decision makers with targeted and useful information, presented in accessible ways. Members of the public can contribute sightings and photos of species and help to build a more complete picture of Australia's biodiversity . Funded by the Australian Government, the Atlas is a collaboration between CSIRO, Australia's national science research agency, and more than 60 biological collections from
Museums and Herbaria, Federal and State Departments, universities and microbial collections.
Note: abstracts are arranged by theme and then by symposium number and then by speaker order within each symposium.
Where a symposium is divided into two sessions (A and B), the abstracts are in chronological order under the one symposium number.
Theme
01: Ecology, environmental change and conservation
Symposium numbers
1, 2, 9–27, 126, 127, 149, 150, 158, 163–167
28–36, 152, 159–163 02: Economic botany including biotechnology, agriculture and plant breeding
03: Genetics, genomics and bioinformatics
04: Physiology and biochemistry
05: Structure, development and cellular biology
06: Systematics, evolution, biogeography and biodiversity informatics
07: Plants in society
37–42, 45–48, 123, 157
49–55, 124, 145, 151, 153, 156
56-71
4–8, 72–122, 128–144, 155, 168–172
3, 177
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Mistletoe specialist frugivores: diligent dispersers or self-serving free-loaders
Watson, DM 1 , Rawsthorne, J 1120
1 Environmental Sciences, Charles Sturt University,
Wagga Wagga, Australia
Parasitic plants necessarily depend on host plants for their water and mineral nutrition, and therefore differ from most plants in the specificity of safe sites needed for germination and establishment. Most groups of parasitic plants use animal-mediated directed dispersal to increase the likelihood of seeds landing on or near suitable host plants. This is exemplified by mistletoes, which attach to single hosts above ground, unlike rootparasites which typically attach to multiple hosts below ground. These specific dispersal requirements have encouraged reciprocal adaptation of mistletoes and birds, and mistletoe specialist frugivores have evolved independently in seven avian lineages. Most of the research conducted on mistletoe dispersal has been restricted to these taxa, their nutritional reliance on mistletoe fruit presumed to confer the status of principal seed dispersers. Recent research has called this into question, however, and the role of mistletoe specialist frugivores as principal dispersers warrants critical reappraisal. In this contribution, we demonstrate that mistletoe specialist frugivores move from infected hosts to other infected hosts, thereby intensifying existing infections far more frequently than initiating new infections (by dispersing fruit to uninfected host). So, even though these birds may remove most seeds, they may reduce individual and population-level fitness of mistletoes by loading hosts with increasing numbers of parasites, lowering overall levels of recruitment. In addition to reviewing recent research on these specialists, we summarise several studies of more generalist foragers that include mistletoe in their diet, demonstrating that they are far more likely to disperse seeds to uninfected hosts and stands. Finally, we compare continental regions with mistletoe-specialist frugivores with other regions
(including Madagascar, New Guinea, New Caledonia,
New Zealand and Europe) where all dispersal is performed by dietary generalists, evaluating the ecological and evolutionary consequences for the plants themselves. Although these coevolved specialists may have been instrumental in the evolution of dispersal syndromes and diversification of mistletoes, we suggest that their dietary specialization has diminished their importance as seed dispersers and determinants of mistletoe distribution, and they may not be the helpful mutualists which they are frequently considered.
Bird–mistletoe interactions with a depleted avifauna: what determines mutualism failure?
Kelly, D 1 , Ladley, J 1 , Robertson, A 2
1 Biological Sciences, University of Canterbury,
Christchurh, New Zealand;
University, New Zealand
2 Ecology, INR, Massey
The Loranthaceae worldwide have some of the most apparently-specialised bird-plant mutualisms both for pollination (such as 'explosive' flowers that cannot open without a pollinator) and for dispersal (with directed dispersal to particular sized host branches). However, the actual level of plant dependence on the mutualism for successful reproduction varies widely among species.
Here we use the New Zealand Loranthaceae to explore the factors which predict sensitivity to mutualism failure.
The question is important in New Zealand because both the mistletoes and their bird mutualists have declined in abundance and some are of conservation concern. For pollination, both Peraxilla spp. and Alepis flavida have ornithophilous flowers, but A. flavida routinely autonomously self-pollinates so is immune to mutualist failure unless there is inbreeding depression (which is known absent in Peraxilla spp., but impossible to test in
A. flavida ). In contrast Peraxilla spp. have explosive flowers and are widely pollen-limited due to a shortage of pollinating birds. However, many flowers are also lost to a native florivorous moth Zelleria maculata , so the relative impacts of pollinaton failure versus flower predation are complex and both vary with habitat fragmentation. For dispersal, all tested loranths have an absolute requirement for passage through a bird gut to allow adhesion to new host branches. However, levels of mutualist service for dispersal seem to be much more adequate than for pollination, even though the same bird species are providing both services. This may reflect a general rule that dispersal is 'easier' than pollination when both are done by vertebrates. There is no evidence for directed dispersal to small branches in the NZ species, so seed losses at the defecation stage are probably very high. Overall the greatest threats to New
Zealand Loranthaceae are introduced herbivores and pollination failure, with dispersal less at risk.
Studies on the biodiversity and host-specific interaction of some parasitic angiosperms (mistletoes) in the Himalayan terrain of Uttaranchal, India
Mondal, AK 1 , Mondal (Parui), S 2
1 Vidyasagar University, Midnapore, India;
Brabourne College, Kolkata, India
2 Lady
An extensive survey of the Himalayan terrain of Nainital,
Almora, Ranikhet, Kausani and adjoining areas of the state of Uttaranchal, India was done to record the parasitic angiosperms growing in this area. Parasitic angiosperms have been found to have significant ecological effects on the plant community structures by altering the competitive balance between host and nonhost species. Six species of parasitic plants or commonly called mistletoes were recorded from this zone. Among these, five were hemi-parasites, which included Cassytha
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filiformis, Dendrophthoe falcate, Loranthus longiflorus,
Phoradendron serotinum, Viscum album , while the sixth member Cuscuta reflexa was a holo-parasite. The present paper reports the distribution, dominance and the host specific interaction of this parasitic species. Viscum album was found to be the most dominating species seriously parasitizing the host Pyrus pashia (Rosaceae), which is an economically important plant. Key words:
Biodiversity, host-specific interaction, parasitic angiosperms.
Soil nutrient accumulation under two species of mistletoe produce different soil patchiness in a semiarid open woodland
Facelli, JM 1 , Watling, J 1 , Binney, A 1
1 The University of Adelaide, Australia
Mistletoes accumulate higher concentrations of nutrients in their foliage than their hosts. Because of their abundance in nutrient-poor systems they may play important functional roles in nutrient cycling. In arid lands they grow on long-lived plants that frequently create patches of high nutrient availability, and may therefore affect the patchiness in nutrient distribution typical of these systems. Here we report a study of patterns of nutrient accumulation under two mistletoes,
Amyema quandang and Lysiana exocarpii , growing on different hosts ( Acacia papyrocarpa and Alectryon oleifolius respectively), in chenopod shrublands of South
Australia. We quantified litter production and litter nutrient concentrations of the four species. We also compared soil nitrogen (N), phosphorus (P), and organic carbon (OC) content in the soils beneath both hosts and mistletoes, and conducted a growth bio-assay using these soils and a co-occurring perennial shrub, Enchylaena tomentosa , that grows almost exclusively under canopies and is known to require high levels of nutrients.
Mistletoes produced up to 6 times more litter than either host, regardless of infection status of the latter. Litter N concentration was similar in A. quandang and its host A. papyrocarpa , but L. exocarpii had slightly higher litter N content than A. oleifolius . In contrast, both parasites had significantly higher litter P concentrations than their hosts. The soil beneath the hosts had higher nutrient contents than soil from adjacent, open spaces, and the soil directly beneath mistletoes had significantly higher
N and P concentrations than soil beneath parts of the host canopies without mistletoes. While there was no difference between the soils underneath the two hosts when they had no mistletoes, the patches underneath the two species of mistletoe were different. The soil underneath A. quandang had less organic carbon and phosphorus, but more nitrogen than the soil underneath
L. exocarpii . These differences were biologically important since biomass of E. tomentosa was higher when grown in soil from beneath L. exocarpii than in soil from underneath A. quandang . Biomass was also higher in soil from under mistletoes than from under canopies of the hosts of from open spaces. Mistletoes contributed both more litter and more nutrients to soil than either tree species, resulting in significant heterogeneity in nutrient availability across this nutrient poor ecosystem.
Enrichment of soils beneath mistletoes resulted in higher growth of a perennial shrub, suggesting that mistletoes could play a significant functional role in nutrient cycling within this semi-arid woodland.
Cytotoxic activity of Mexican Mistletoe (
Cladocolea loniceroides
)
Serrano Maldonado, MJ
,
1 , De la Paz Perez Olvera, C 1
Soriano Santos, J 1
,
1
Guerrero Legarreta, I 1
Universidad Autónoma Metropolitana, Itzapalata,
Mexico
Mistletoe, as a parasitic plant, grows attached to and within the branches of trees and shrubs. Cladocolea loniceroides seed, a Mexican endemic mistletoe, can be spread through the feces of birds moving from tree to tree throughout the country. It significantly infests them and can be a serious pest in forest landscapes and cities’ gardens. Its impact is rather negative causing great tree mortality. The most effective way to control it is to prune out infected branches, but mistletoe is useless. Thus, the aim of the study was to investigate both in vitro and in vivo antioxidant activity of C. loniceroide s extract, as obtained of leaves, stems or fruits (L, S, F), so as to observe the in vitro cytotoxic activity against breast cancer cells with the purpose of using this plant to obtain an added-value product. Extracts from L, S or F were obtained by using methanol, water or saponification reaction. Then each extraction was assayed to assess total polyphenolics by means of the Folin-Ciocalteu reagent
(fruit>leave>stem: 189.5 ± 4.9; 93.33± 0.94 and 74.53 ±
0.95 mg gallic acid (GA)/g, respectively). Antioxidant activity (AOxA) was evaluated by DPPH•;
ABTS/laccase; metal ions chelating activity and reducing power methods. L, S or F extracts showed up to 90.27 ±
0.13 % DPPH inhibition; similar results were observed when AOxA was evaluated by ABTS/laccase system.
The reducing power of all extracts was higher than that of reduced glutathione, which was used as control. On the other hand they exhibited a lower Fe2+ chelating activity than that of EDTA. Lipoperoxidation induced in rats was inhibited when those animals were fed with a fruit aqueous extract (FAE) and there was not significant differences (p<0.05) when compared to the rat control group. Later on, FAE showed a citotoxic activity on
ZR75 breast cancer cell line (LD50= 0.1 mg GA/mL).
Therefore C. loniceroides may be processed to obtain an antioxidant standardized extract which might be useful in treating breast cancer.
Diversity and host-specificity of invertebrates inhabiting an Australian mistletoe and its host eucalypts
Burns, AE 1,2 , Watson, DM 2 , Cunningham, SA 3
1 Monash University, Clayton, Australia;
University, Australia;
2 Charles Sturt
3 CSIRO Ecosystem Sciences,
Canberra, Australia
Parasitic plants, such as mistletoes, are important components of tree canopies, providing an alternative habitat and resources for a range of animals. However, the diversity and host-specificity of invertebrates on mistletoes and their host-plants has received little attention; thus over-looking the contribution of these component communities to the biodiversity of tree
18

canopies. This study investigated the species composition and host-specificity of invertebrates inhabiting hemiparasitic box mistletoe, Amyema miquelii (Lehm. ex
Miq.) Tiegh. in Eucalyptus trees. Insects, spiders and mites (i.e. arthropods) were sampled from box mistletoe and its host eucalypt trees in remnant woodlands in temperate Australia. The same orders of arthropods were found on box mistletoe and the host eucalypts but the density of arthropods was greater on the eucalypts than the mistletoe plants. Two groups of arthropods in different trophic groups were investigated in more detail.
The species composition of the herbivorous psyllid
(Hemiptera: Psylloidea) insect assemblages (21 species total) differed significantly between the plant species, but the spider (Araneae) assemblages, which consisted of 42 species in total, were similar in species composition between the mistletoes and eucalypts. Elucidation of tourist species revealed less than 1% similarity in community composition between the psyllid assemblages on the mistletoes and eucalypt trees. In comparison, the spider assemblages displayed 40% similarity in community composition between the mistletoes and eucalypts. Therefore, the greatest contribution of mistletoe plants to canopy arthropod diversity was at the lower rather than higher trophic levels of arthropods.
Mistletoe plants in general are likely to support different herbivorous insect assemblages compared to their hostplants due to differences in plant physiology. By comparison, predatory arthropod assemblages are likely to be similar in species composition between mistletoes and their host-plants due to similarities in habitat structure, but differ in abundance due to variation in prey densities on the plants.
A fresh look on desiccation-tolerant vascular plants from an anatomical perspective
Korte, N 1 , Porembski, S 1
1 University of Rostock, Mecklenburg–Vorpommern,
Germany
Rock outcrops such as inselbergs can be found in all climate zones but are especially abundant in the tropics.
They form a center of diversity for desiccation-tolerant plants which is a very rare trait among higher plants
(1.500 spp.), especially angiosperms (300 spp.). The physiological mechanisms of desiccation tolerance have been the subject of many research efforts but comparative anatomical and morphological studies under the aspect of poikilohydry are lacking. Therefore, desiccation-tolerant Cyperaceae (A frotrilepis pilosa,
Microdracoides squamosus ), Velloziaceae
( Barbaceniopsis castillonii, Vellozia andina, Xerophyta sp.,), Borya nitida (Boryaceae), Tripogon spicatus
(Poaceae) and the dicotyledonous Myrothamnus flabellifolius and M. moschatus (Myrothamnaceae) were chosen to compare leaf anatomical traits (turgescent and dry). Main objectives included the determination of important anatomical structures, which facilitate the survival of complete desiccation, and the question whether an ‘anatomical desiccation-tolerant plant syndrome’ exists. It could not be confirmed that desiccation tolerance, leaf anatomy and morphology are necessarily related, but differences among and between monocotyledonous and dicotyledonous species were abundant: leaves of Cyperaceae and Velloziaceae folded symmetrically due to the existence of bulliform cells above the midrib, which collapse during desiccation and facilitate the folding of the leaf. All monocotyledonous leaves showed a high amount of vascular, sometimes marginal, sclerenchyma and single sclerenchymatic islands in the mesophyll as stabilizing elements. The leaves of M. flabellifolius and M. moschatus had less to no xeromorphic characteristics, were mainly parenchymatic, and folded accordion-like. Summarizing, it can be stated that (leaf) anatomical traits are a piece in the puzzle of desiccation tolerance but cannot be generalized across species or species groups; similar traits can also be detected in other xeromorphic species.
The adaption of desiccation-tolerant plants to subsequent de- and rehydration is not only to be found in their anatomy and morphology but naturally on the physiological level as well.
Hybridization and interespecific gene flow in
Pitcairnia
(Bromeliaceae): the maintenance of species cohesion among isolated populations adapted to rock outcrops in southeastern Brazil
1
Palma-Silva, C
Cozzolino, S 4
1 , Wendt, T 2
, Lexer, C 5
, Pinheiro, F
, Barbarà, T 6
Instituto de Botânica, São Paulo, Brazil; 2
1 , Fay, M 3 ,
Departamento
3 de Botânica, CCS/IB/UFRJ, Rio de Janeiro, Brazil;
Jodrell Laboratory, Royal Botanic Gardens Kew,
4 Richmond, Surrey, UK; Dipartimento di Biologia
Strutturale e Funzionale, Complesso Universitario di
Monte S. Ângelo, Università degli Studi di Napoli
Federico II, Napoli, Italy; 5 Unit of Ecology and
Evolution, Dept of Biology, University of Fribourg,
Switzerland; 6 University of Fribourg, Switzerland
The reasons for high rates of speciation promoting species richness and endemism of rock outcrops are not entirely understood. The isolation among rock outcrops together with their long persistence have contributed to high levels of differentiation of endemic species, that were crucial in speciation processes in these habitats.
High population differentiation among rock outcrops indicates that they could be comparable to oceanic islands on studies of evolutionary processes.
Bromeliaceae has several genera adapted to Neotropical rock outcrops. Recently bromeliads attracted research interest in population connectivity and its consequences for speciation and species cohesion in isolated environments. Rock outcrops insular features allow us to use them as models systems for testing the role of low intra-specific gene flow and high and continuous interspecific gene flow during plant speciation. Here we studied several populations four sympatric Pitcairnia species adapted to rock outcrops using plastidial and nuclear microsatellite loci. Our aim was to understand the relative role of the gene flow (intra- vs. inter-specific) and the reproductive barriers (prezygotic vs. postzygotic) during speciation processes in rock outcrop bromeliads species. Extensive haplotype sharing and nuclear admixture analyses indicated inter-specific gene flow
19

among species (hybridization and introgression). Nuclear migration rates and highly geographically structured haplotype sharing support that intra-specific gene flow is low among populations (< 1 Nem per generation), likely due to restricted seed dispersal and long term interspecific gene flow during bromeliad adaptive radiation on rock outcrops. Patterns of nuclear genomic diversity and admixture indicate that both pre and postzygotic barriers, although incomplete, can potentially contribute to the reproductive isolation, limiting gene flow between species. Traits that contribute to assortative mating such as differences in flowering time, pollinator specificity, and mating systems (selfing vs. outcrossing rates) are important pre-zygotic barriers. Post-zygotic barriers, such as nuclear-nuclear and cyto-nuclear incompatibilities are also of great importance in shaping reproductive isolation among species. Hybridisation and introgression could contributes to bromeliad biodiversity by enriching local gene pools on rock outcrops. Thus, inter-specific gene flow should be important for these plants, especially because intra-specific gene flow is low.
Consequently, rock outcrops with multiple sympatric bromeliads deserve special attention in conservation programs. Finantial support: FAPESP; CNPq; CAPES
Evolution of rock outcrop endemics in eastern Brazil: a phylogenetic case study in the saxicolous bromeliad genus
Orthophytum
Beer
Louzada, R 1 , Schulte, K 2 , Silvestro, D 3 , Wanderley, M 4 ,
1
Zizka, G 5
Dept of Botany, Biosciences Institute, University of São
Paulo & Núcleo de Pesquisa Curadoria do Herbário,
Instituto de Botânica, São Paulo, Brazil; 2 Australian
Tropical Herbarium, James Cook Unversity, Cairns,
Australia; 3 Dept of Botany and Molecular Evolution,
Senckenberg Research Institute & Biodiversity and
Climate Research Centre, Frankfurt, Germany; 4 Núcleo de Pesquisa Curadoria do Herbário, Instituto de
Botânica, São Paulo, Brazil; 5 Dept of Botany and
Molecular Evolution, Research Institute Senckenberg &
Dept of Diversity and Evolution of Higher Plants,
Institute of Ecology, Evolution and Diversity, Goethe-
University, Frankfurt/Main, Germany
Eastern Brazil harbors a great variety of rock outcrop formations, most notably the inselbergs within the
Atlantic Rainforest domain and the campos rupestres of the Espinhaço Range. These rock outcrops are characterized by a highly specialized flora adapted to high insolation, periods of severe drought, and different substrates, and are rich in endemics and microendemics.
Molecular phylogenetic studies of Brazilian rock outcrop plants elucidating the factors contributing to their species diversity and to the evolution of endemism are still scarce. Here we present an AFLP study of the saxicolous bromeliad genus Orthophytum (60 spp.) that diversified extensively along rock outcrops of eastern Brazil and which comprises a high proportion of species endemic to inselbergs and campos rupestres. More than 50% of the species were described in the last two decades and assigned to the two major morphological complexes and six sub-complexes that are discerned within the genus based on morphological characters, however no molecular phylogenetic study has been conducted so far to infer relationships. In the AFLP study 48 species and
20
101 samples were included, covering the entire geographical range of the genus, and investigated with 12
AFLP primer pairs yielding 4679 characters. The resulting binary matrix was analyzed using Neighbor
Joining, Maximum Parsimony, and Bayesian methods to infer phylogeny. A NeighborNet analysis as implemented in the program SplitsTrees was conducted to detect possible conflicting signals within the data set as can be caused by phenomena like hybridization and introgression. A Bayesian clustering algorithm as implemented in the software Structure was used to assess the main genetic structure within Orthophytum and possible admixture between main clusters. Detailed distribution data was collected from herbarium specimen at 26 herbaria (ALCB, ASE, B, BHCB, CEPEC, ESA,
HAL, HB, HEID, HUEFS, IPA, K, MAC, MBM,
MBML, NY, P, R, RB, SP, SPF, UEC, UFP, VIC, W,
WU) to assess ecological niche parameters of the taxa via the WorldClim database. The data were used to model the potential distribution of the taxa and to infer phylogenetic shifts in ecological niches. The phylogenetic reconstructions obtained are well resolved showing six monophyletic groups within Orthophytum sensu stricto: the foliosum, mello-barretoi, magalhaesii, glabrum, estevesii and saxicola clades. The two traditionally recognized major sub-complexes, the pedunculate and the sessile inflorescence group, are clearly paraphyletic. Based on the phylogenetic evidence, different hypotheses regarding the historical biogeography of Orthophytum (migration routes, past refugia) are tested and the importance of past range expansions and contractions during Cenozoic climatic oscillations for the diversification of the genus along the rock outcrops of eastern Brazil are discussed.
Endemism and speciation on rocky outcrops of
1
Western Ghats, India
Janarthanam, Malapati 1
Goa University, Goa, India
The Western Ghats of India along with Sri Lanka is one of the 34 biodiversity hotspots of the World. The
Northern Western Ghats and narrow plains between West
Coast and Central & Northern Western Ghats are characterized by the occurrence of lateritic outcrops that form plateaus and table lands. They provide unique habitat to monsoon flora between the months of June and
October. Several hundred herbaceous species show temporal succession and complete their life cycle during this period. A good proportion of these species are endemic to W. Ghats – some very narrow in their distribution and some others known only from their type localities. Most of these endemics are habitat specific, i.e. they are seen only on lateritic outcrops. The major and prominent groups with high proportion of endemics are
Andropogoneae (Poaceae), C eropegia (Apocynaceae),
Utricularia (Lentibulariaceae), Smithia (Fabaceae),
Impatiens (Balsaminaceae) and Eriocaulon
(Eriocaulaceae). Among grasses, endemic species-rich C
4 genera such as Ischaemum and Glyphochloa are extensively distributed. Interestingly all these endemic species rich taxa belong to the terminal higher level taxa of respective clades. Close relatives of these endemic taxa extend their distribution to Africa and Australia.
Most of their relatives also show high degree of

endemism in these distant lands. Recent sporadic floristic studies on lateritic outcrops brought forth several new species to the fore during the past two decades. Most of these endemic taxa show sympatric distribution.
Occasionally some populations show intermediate or overlapping characters. These facts suggest that endemic taxa are fast evolving and speciation is taking place at a rapid rate. The driving force behind the speciation on these rocky outcrops seem to be hybridization and polyploidy. Heterogeneity in the habitat at finer level appears to be another factor. This heterogeneity is seen in the form of patchy distribution of some species even in small areas though others are dominant throughout the area. The heterogeneity is also evidenced from the physical parameters such as the exposed hard lateritic rocks, amount of lateritic gravel, thickness of soil (if any) covering the rocks, size and depth of puddles (rock pools) on lateritic rocks and ultimately precipitation pattern. These factors ultimately decide the moisture regime and water availability, especially during interprecipitation events. These combined factors appear to be responsible for high endemism and speciation. However, these rocky outcrops have not attracted sufficient attention from the botanists thus leaving lot of gaps in understanding of the flora and the processes that drive the speciation and endemism. On the other hand, these habitats with rich endemic species and fast evolving flora are under threat due to habitat degradation warranting urgent attention of botanists and conservationists.
The challenges of determining success for reintroductions of threatened long-lived plants
Monks, L 1 , Dillon, R 1 , Coates, D 1
1 Dept of Environment and Conservation, Bentley,
Australia
Reintroduction to new, current or extinct locations is increasingly becoming an important action to recover some of our most threatened plant species. The challenge is to devise appropriate criteria which allow us to assess the success of the reintroduction program. Many reintroduction programs monitor establishment, growth, reproduction and recruitment over multiple generations before attempting to determine whether the population is viable and self sustaining. Only when a reintroduced population is shown to be viable and self-sustaining should it be taken into consideration when determining the conservation ranking of the taxon. However for reintroductions programs of long lived woody shrubs determining success is particularly challenging given the timeframes over which these taxa reproduce and recruit new generations. Since 1993 the Department of
Environment and Conservation, the government agency responsible for managing all flora in Western Australia, has commenced reintroduction programs for 52 longlived woody plant taxa. Just five of these reintroduced taxa have recruited a second generation. In determining success long-term monitoring of survival, growth and reproduction is being undertaken on reintroduced populations and compared to natural populations.
However, clearly there needs to be some surrogate method by which long term success can be predicted in the timeframes in which a management agency is required to operate. Population viability analysis models and mating system analysis are likely to be useful tools in predicting long term population trends.
Community ecology and reintroduction of
Tigridiopalma magnifica
, a rare and endangered herb
Ren, H 1 , Zeng, S 1
1 South China Botanical Garden, Chinese Academy of
Sciences, Guanghzhou, China
Tigridiopalma magnifica , a perennial herb and the only species in the genus Tigridiopalma (family
Melastomataceae), is rare and endemic to China. Twelve sites with populations of T. magnifica have been identified (1 extinct, 11 extant). T. magnifica only grows on the surface soil of stone walls or rocks under the canopy of secondary forests and plantations and has no specific associated plant. Canopy closure, soil water content, and the distance to the closest stream are major factors affecting T. magnifica distribution. We reintroduced T. magnifica tissue-culture plantlets into three field sites; one site was near an original distribution area, one was 11.5 km away, and one was 400 km away.
After 11 months, the survival rate ranged from 40 to
58%, but survival was higher and plantlet crowns were larger at the site near the original distribution area than at the other two sites. The combination of advanced propagation techniques and ecological restoration could enable successful reintroduction and conservation of T. magnifica and other rare and endangered plants.
Conservation strategies for rare and endangered medicinal and aromatic plants
Jamzad, Z 1
Mohebi, J 1 ,
, Charkhchian, MM 2
Pourmirzaii, A 4 , Hasani-Nejad, M 1
Maroofi, H 5 , Safikhani, K 6
, Hatami, A
, Araghi, MK
, Ahmadi, S 7
3
1
,
,
1 Research Institute of Forests & Rangelands, Tehran,
Iran; 2 Research Centre of Agriculture and Natural
Resourses, Ghazvin, Iran; 3 Research Centre of
4
Agriculture and Natural Resourses, Shiraz, Iran;
Research Centre of Agriculture and Natural Resourses
Krman Iran; 5 Research Centre of Agriculture and
Natural Resourses, Kordestan, Iran; 6 Research Centre of
7
Agriculture and Natural Resourses, Hamadan, Iran;
Research Centre of Agriculture and Natural Resourses,
Lorestan, Iran
The increasing uses of medicinal and aromatic plants which in most cases are harvested directly from the nature has caused sever damages to this important plant groups in Iran. The country supports around 8000 species of spermatophytes belonging to 150 families and is one of the major centers of endemism in Irano-Turanian phytogeographical region. Twenty two percent of these species are categorized as endemics. There are narrow endemics among some of the currently used medicinal and aromatic plants. In this project we screened 20 perennial species of medicinal and aromatic plants with an emphasis on endemic species. Their natural habitats, reproduction and soil seed banks were studied. The
21

project was designed with the following objectives: (1)
Propagation and cultivation of them in National
Botanical Garden of Iran; (2) keeping them as a germplasm and conserving them in a controlled environment; (3) reintroducing them to their natural habitats and recreating their natural populations. These studies revealed that the seeds of some of them are scant in soil seed banks of their habitats and living stands are few with little or no regeneration. Factors that have caused this situation include harvesting the plants before seed setting, lack of seed production as a result of physiological disorders, pest infestation, and habitat disturbance or draught. Among the aforementioned factors, inaccurate harvesting and draught were the most important ones in our study group. We thus collected the seeds of selected species from their natural habitats, germinated them in the greenhouse under controlled conditions providing them with sufficient moisture through mist propagation units. The seedlings were then transferred outdoors in the garden and their phenological behaviors were recorded. The seeds of cultivated specimens were then collected and saved for replanting in the garden and in their original habitats. The seedlings of new grown specimens will be transferred to their original habitats to give them a chance to produce new populations. The cultivation experience of
Dracocephalum kotschyi, Gontscharovia popovi; Nepeta pogonosperma; N. rvularis; N. assurgens; Origanum vulgare; Thymus daenensis; T. kotschyanus; T. persicus;
T. pubescens; Salvia oligophylla; Ziziphora clinopodioides showed their remarkable adaptability to new ecological conditions different from their own.
The study of population genetics in the endangered species
Pleomele hawaiiensis
in Hawaii
Lu, P-L 1
1 Botany Dept, University of Hawaii at Manoa, USA
The Hawaiian Archipelago includes various endemic species that are the result of speciation subsequent to isolation from source populations. Pleomele hawaiiensis
Degener & Degener is an USA federally listed endangered species by the US Fish and Wildlife Service
(USFWS) since 1996 and listed in the International
Union for the Conservation of Nature and Natural
Resources (IUCN)’¦s Red List of threatened species since
1997. The main reasons for its rarity might be due to habitat loss by development encroachments and the introduction and spread of invasive animals. There is no previous research about its ecological, genetic, and physiological biology. Based on the flower and seed structure and morphology, bees are potential pollinators and birds are the possible seed dispersal candidate.
According to Wagner’s classification, there are six endemic Pleomele species in Hawaii Islands. The other species ( P. aurea (H.Mann) N.E.Brown, P. auwahiensis
St John, P. fernaldii St John, P. forbesii Degener, and P. halapepe St John) are distributed throughout the other high islands of the Hawaiian archipelago. Only the single species P. hawaiiensis is recognized as occurring on the
Big Island. P . hawaiiensis is endemic to Hawai’i in the state of Hawaii in six to eight subpopulations totaling approximately 300 individuals in Puuwaawaa, the
Kaloko/Kaloao area, the Kapua/Kahuku area, and on
Holei Pali (located in the Hawai¡¦i Volcanoes National
22
Park). Currently, no young seeding exists around and within those populations. Typically, they grow in dry forest on lava fields. Because of its rarity and small population sizes, it may go extinct or become more severely depressed quickly in few decades if no proper preservation actions are taken. In the past ten years, the
US government tried to recover this species by artificial planting in the conservation areas in Hawai’i. My research goal is to perform a molecular genetic analysis of the DNA polymorphism in populations of Pleomele hawaiiensis aiming to determine the level of their genetic structure within this species. RAPDs and ISSR results indicate the remaining wild population has lower genetic diversity than the artificial planted population, and shows this species faces severe genetic bottleneck. This species may have no longer long distance seed dispersal and lack efficient pollinators. The data supports other researches that genetic deterioration may result from decreasing population size and populations tend to have lower genetic variation, especially in rare plant species. This study points out the reintroduced plants can have better genetic diversity for form a more healthy population in the endangered plant species if the seeds resource are collected from different populations. That will help the endangered species recovery in reality. A better understanding of population genetic structure will benefit the conservation work and explain the species’ evolutionary history. Selecting seeds from the most different genetically diverse individuals from the genetic data of population structure is important to preventing the endangered plant species toward towards extinction from the wild.
Experimental establishment in situ of
Dactylanthus taylorii
(Balanophoraceae), an endangered holoparasitic flowering plant in New Zealand
Holzapfel, S 1 , Dodgson, J 2
2
1 Dept of Conservation, Hamilton, New Zealand;
Kawhia, New Zealand
Holoparasitic flowering plants provide considerable challenges for conservation management due to their absolute dependency on a host plant, and because many aspects such as germination, host attachment and growth are often not well understood. We present results from the first successful, quantified field trial to establish from seed populations of dactylanthus ( Dactylanthus taylorii ), the southern-most member of a family of rootholoparasitic angiosperms (Balanophoraceae).
Dactylanthus is an acutely threatened species endemic to
New Zealand, growing underground as a perennial tuber attached to the root of native host trees and shrubs.
Nectar-rich inflorescences break through the forest floor, where they are pollinated by a ground-foraging endemic bat. The resulting copious small seeds form long-lived, dense local seed banks and germinate independent of host presence. Browsing of inflorescences by introduced mammals is limiting the species’ recruitment and has led to its disappearance over 96% of its pre-human distribution range. Being able to establish new populations by sowing would provide an important tool for the recovery of the species. A known quantity of
Dactylanthus seed was sown into twenty-four field plots at four sites differing in host species dominance, host age and canopy closure. Two sowing methods were used at

each site, replicating individually dispersed seeds and an entire buried infructescence, respectively. Over ten years plots were monitored annually at the peak of flowering time. Successful establishment was first confirmed in two plots four years after sowing; six years later
Dactylanthus was present in twenty-two plots. Plants generally appeared healthy and were still growing in number and size at the conclusion of the trial. Average and maximum inflorescence numbers per plot were high compared to protected wild populations, though successively smaller increases each year indicated that the population was approaching its maximum flowering capacity. The two sowing methods resulted in similar number of successful plots overall, while broad sowing resulted in a greater number of inflorescences and, by proxy, individual plants per plot. Establishment and inflorescence numbers were similar at sites with different host species and host ages, but were lower for the open canopy site. Female inflorescences strongly outnumbered males at all four sites and in every flowering year, though the proportion of male inflorescences increased each season. This is in stark contrast to wild populations, which are usually characterised by an over-abundance of plants producing male inflorescences. These results, and the rare occurrence of hermaphroditic inflorescences in a species usually regarded as dioecious, are explored as possible evidence of previously undetected monoecy, subdioecy, or sex-switching of individuals. While the success of this study has direct applications for the conservation management of Dactylanthus , results also provide important new insights into the biology and ecology of the species and holoparasitic flowering plants in general.
Conservation and reintroduction of four threatened trees in southwest China
Sun, W 1
1 Kunming Botanical Garden, Kunming Institute of
Botany, Chinese Academy of Sciences, China
Magnolia sinica ( Manglietiastrum sinicum ), M. phanerophlebia and Michelia coriacea from
Magnoliaceae are critically endangered endemics to southwest China’s Yunnan Province. Among the three extant species from genus Trigonobalanus (Fagaceae), T. doichangensis is a globally endangered plant native to
North Thailand and South Yunnan of China (firstly recorded in 1981). M. sinica, M. phanerophlebia,
Michelia coriacea and T. doichangensis are scientifically important and ornamentally valuable, and they are facing an extremely high risk of extinction in the wild.
Kunming Botanical Garden (KMBG) under Kunming
Institute of Botany, CAS, has carried out comprehensive studies on the population/community ecology, reproductive biology/ecology, and cytological and genetic variations of the four threatened tree species since 2001. Based on good understanding of the threats, scientifically sampled seed collections and propagation from seeds, a great number of seedlings/saplings of M. sinica, M. phanerophlebia, Michelia coriacea and T. doichangensis have been well conserved ex situ at
KMBG. Meanwhile, actions of reintroduction / reinforcement for the four trees to the semi-natural areas which they formerly occurred or to the individually scattered current habitats have been undertaken since
2007, in cooperated with Botanic Gardens Conservation
International (BGCI), Fauna & Flora International (FFI), the Forestry Department of Yunnan Province (FDY) and the local forestry bureaus or nature reserves. The presentation is trying to summarize all the scientific achievements and conservation activities on M. sinica,
M. phanerophlebia, Michelia coriacea and T. doichangensis , and is aiming to document these valuable cases for globally threatened trees conservation.
Laser-based trace gas detection with applications in biology and agriculture
Harren, FJM 1
1 Radboud University, Nijmegen, The Netherlands
There is a strong growing interest to develop laser systems (quantum cascade lasers, optical parametric oscillators) in the mid-infrared wavelength region for trace gas detection with applications in environmental sciences, biology, agriculture and medical sciences.
Nowadays, such lasers provide output at relatively high power and narrow linewidth and emit at any desired wavelength within the infrared wavelength range 2.5 to
12 micrometer. Accurate detection of specific gases becomes into reach thanks to the infrared fingerprint absorption spectrum of molecular gases in this wavelength region and the exact tuning capabilities of lasers. When the lasers are combined with sensitive spectroscopic techniques, such as photoacoustic spectroscopy or optical cavity enhanced spectroscopy, gases can be determined extremely sensitive under atmospheric conditions. Examples will be given on the detection of the important plant hormone ethylene, detection of fermentation products of fruit (acetaldehyde and ethanol) and the detection of NO under pathogen attack. In addition, we will discuss whether plants do produce methane, using C-13 enriched plants and detecting carbon-13 methane via infrared spectroscopy.
In contrast to our expectations we could detect no significant methane emissions.
Comprehensive GCxGC-TOFMS analysis of plant
1 volatiles: a newcomer in the plant signaling toolbox?
Gaquerel, E 1
Max Planck Institute for Chemical Ecology, Jena,
Germany
Metabolomic tools provide novel opportunities for system-wide analysis of complex functions involving single chemical compounds or structure groups. Volatile compounds emitted by plants mediate important ecophysiological functions such as attraction of pollinators to flowers or the recruitment of natural enemies of herbivores. Plant volatiles are products of a complex network of biochemical pathways, which, although well mapped from a biochemical point of view, remains only partly understood with regard to its physiological and
23

genetic regulation. Our group uses comprehensive twodimensional gas chromatography coupled to time of flight mass spectrometry (GCxGC-TOFMS) to track herbivory-induced reconfigurations of the volatile metabolome of Nicotiana attenuata, a wild tobacco species. Changes in volatile components must be extracted from the thousands of molecular fragments that constitute a typical GCxGC map. To that end, data-sets are first filtered and aligned by similarity matching with a synthetic sample reference and compared using univariate and multivariate techniques. The use of this exploratory technique to elucidate the interactive control exerted by herbivore salivary components and plant signaling transduction pathways on volatile emissions will be discussed.
1
Ethylene and cell death signaling in plant and algal cells
Woltering, EJ 1 , Iakimova, ET
2
Wageningen University, Wageningen, The Netherlands;
Institute of Ornamental Plants, Sofia, Bulgaria
Programmed cell death (PCD) is a process where cells or tissues are broken down in an orderly and predictable manner, whereby nutrients may be re-used by other cells, tissues or plant parts. Currently two main types of PCD are recognised in plants: autophagic and non-lysosomal
PCD; the latter is often referred to as apoptotic-like PCD.
True apoptosis, which is accompanied by digestion of
(parts of) the dying cells in the lysosome of other cells, has not (yet) been observed in plants. Dying cells of either type of PCD show characteristic features such as shrinkage of the cytoplasm and compaction and/or fragmentation of the nucleus. In plants PCD is involved in many developmental and formative processes including xylogenesis, reproductive events, formation of leaf perforations and in the senescence of flowers and leaves. PCD is involved in the response to adverse environmental conditions such as chilling and heat stress, exposure to toxic chemicals or UV radiation and as a result of oxygen depletion. In addition, PCD is involved in the (resistance) reaction to pathogens such as bacteria and fungi. In many systems ethylene plays a role in PCD, although it may not be the primary inducer of cell death.
The involvement of ethylene in cell death in plant and algal cells will be discussed.
Nitric oxide and ethylene interactions in plant disease development and resistance
Mur, LAJ
Harren, F 2
1 , Sivakuruman, A 1 , Hall, MA 1 , Cristescu, S
1 Abersytwyth University, Wales, UK; 2
2 ,
Radboud
Univerisity, Nijmegen, The Netherlands
The gaseous hormones nitric oxide and ethylene have been shown to play important roles in responses to plant disease development and in disease tolerance. To gain further insights into the roles of these hormones trace gas detection methods were employed based on
Photoacoustic and Quantum Cascade lasers (PA and
QCL, respectively) were used for on on-line, in planta measurements. To strengthen the power of these analyses, they included transgenic and mutated plant
24 lines which were altered in either the perception of generation of NO or C
2
H
4
. Initially focusing on tobacco
( Nicotiana tabacum ) and Arabidopsis NO and ethylene production during attack by Pseudomonas syringae were assessed. Gaseous hormone production during interactions leading to disease and resistance linked to the formation of a localised cell death known as the
Hypersensitive Response (HR) were compared. Through the use of lines with modulated nitrate reductase (NIA) activity, this was confirmed that nitrate reductase to be the major source of NO during responses to pathogens.
By far the highest rates of NO production were to be observed during HR and this was shown to be essential for full plant resistance to disease. Equally, detoxification of NO by plant-encoded flavohaemoglobins influenced the speed and extent of HR formation. Examination of
C
2
H
4
production during the HR indicated that the biphasic generation pattern was greatly influenced by
NO. Perturbation of the ethylene biphasic wave compromised HR-mediated resistance against pathogens.
The role of NO and C
2
H
4
was examined in tobacco
(Solanum esculentum) following challenge by the economically important pathogen Botrytis cinerea. This is a necrotrophic pathogen, causing plant cell death as part of its pathogenic process. With this interaction considerable rates of NO and ethylene production were observed, greater than were previously observed with the
Pseudomonas syringae elicited HR. This undoubtedly reflects the infection strategy of Botrytis and demonstrated that NO/ C
2
H
4
production in this case is disease associated and thus, are not invariably associated with resistance. Examination of NO- or C
2
H
4
insensitive tomato mutants showed that disease development was reduced, confirming their roles in disease development.
The regulation of and effects of these NO /C
2
H
4 interactions are currently being investigated but these studies demonstrate how novel insights can be gained when genetic approaches are integrated with careful – in this case, gaseous, hormone measurements.
Airborne signaling between undamaged plants – effects on plants and insects
Glinwood, R 1 , Ninkovic, V 1 , Pettersson, J 1
1 Swedish Univ. Agricultural Sciences, Uppsala, Sweden
Plants damaged by herbivores or pathogens release volatiles active in defense signaling between or within plants, but undamaged plants may also engage in chemical interactions. We present results that contribute new perspectives on allelopathy and use of volatile cues by plants and insects. In a system consisting of different plant species and different barley genotypes we show that airborne signaling affects plant biomass allocation (1), reduces attractiveness to an insect herbivore (2, 3) and affects interactions with the herbivore’s predators (4). In barley, the inducing and responding capacity is shown to be genotype-related. Volatile profiling shows differences in the volatile blends released by different barley genotypes. Increased understanding of chemical interaction between undamaged plants may give a new outlook on the ecology of mixed genotype cropping and weed-crop interactions and their role in sustainable crop production (5). (1) Ninkovic 2003. (2) Glinwood et al.
2007. (3) Glinwood et al. 2004. (4) Glinwood et al. 2009.
(5) Ninkovicet al. 2009.

Tracing hidden herbivores by analyzing plant volatile emissions
Van Dam, NM
Harren, FJM 2
1 , Crespo, E
, Cristescu, SM
2
2
, Laska, A 2 , Hordijk, CA 3 ,
1
2
Radboud University/IWWR, Nijmegen, The Netherlands;
Radboud University/Trace Gas Lab, Nijmegen, The
Netherlands; 3 Netherlands Institute for Ecology,
Wageningen, The Netherlands
Plants are attacked by many different herbivores, both above and below the ground. It is relatively easy to see whether, when and how much plants have been damaged by above ground herbivores. Even when the herbivores have left the plant, the remaining damage can be used to identify the insect species that has been feeding. This is more difficult to assess in the case of belowground feeding root herbivores. Here we illustrate how trace gas analyses can be used to trace infestations by the cabbage root fly ( Delia radicum ). The larvae of this fly species are a serious pest damaging roots of many different crucifer species. Their feeding activities reduce the yield and market value of rape seed, broccoli, Brussels sprouts and other cruciferous crops. Previous studies have shown that the headspace of root fly infested Brassica nigra plants contains specific volatiles. One of these compounds, dimethyldisulfide (DMDS), attracts natural enemies of D. radicum in the field and repels aboveground biocontrol organisms. On-line Proton-
Transfer Mass-Spectrometry (PTR–MS) analysis showed that DMDS emissions, and those of the structurally related methanethiol and dimethylsulfide (DMS) from the roots indeed increased significantly 12–16 hours after root fly infestation. Additionally, infested plants showed markedly increased emission rates of m/z 60. The increased emission of m/z 60 was much faster than the induction of sulfides, starting within 4 h after infestation.
Interestingly, m/z 60 emissions ceased when larvae pupated, thus making it a marker for actively feeding larvae. Artificial wounding of the roots briefly increased the emission rates of m/z 60 as well, whereas root fly larvae alone did not produce an m/z 60 signal. This suggests that m/z 60 is produced by the plant and likely is a product of glucosinolate conversion by myrosinase and other enzymes involved in this process. Analyses of pure (iso)thiocyanates revealed that ethyl and allyl
(iso)thiocyanates both yield m/z 60 as a main fragment in the PTR-MS. Eventually we identified m/z 60 as thiocyanic acid using a pure reference in the PTR-MS.
Our results show that trace gas analyses of root emitted volatiles can be used to assess which plants are infested with actively feeding root fly larvae. Trace gas analyses thus may be applied in quarantine procedures aiming to assess whether imported plants or produce is infested with cryptically feeding pests. Additionally, these results are valuable for plant breeders aiming to select crops with enhanced attractiveness to natural enemies of D. radicum .
Progress in implementing the Global Strategy for
1
Plant Conservation
Wyse Jackson, P 1
Missouri Botanical Garden, St Louis, USA
In 2002 the international community adopted the Global
Strategy for Plant Conservation through the United
National Convention on Biological Diversity (CBD). The
Strategy and its different elements quickly became well known worldwide and highly influential in helping to shape the programmes and policies of many plantorientated institutions and organisations. Its objectives and the 16 outcome-oriented targets it incorporated, to be achieved by 2010, became major drivers for many national and international plant conservation initiatives too. However, after eight years of its operation, when the
GSPC has being renewed by the CBD, with a set of 16 updated targets, it is time to review progress in its implementation to help evaluate what new steps are need in order to achieve its objectives for the second period of the GSPC up to 2020. This paper will provide an overview and evaluation on whether the Strategy is achieving what it set out to do. It will also consider the following questions. Are countries fully engaged in its implementation? What new measures will be needed to ensure that the targets are met by 2020? Issues that are hindering progress in its implementation will also be reviewed, including lack of data, tools and technologies; limited funding; poor sectoral coordination and limited institutional capacity and capabilities, as well as lack of appropriate policies and legal frameworks at the national level. The ways in which individual botanical institutions can seek to address these shortcomings will be discussed.
The existing and future roles of the Global Partnership for Plant Conservation (GPPC) in supporting the GSPC will be highlighted.
1
A working list of known plant species – progress and lessons learned for the GSPC
Paton, A 1 , Nic Lughadha, E 1
Royal Botanic Gardens, Kew, UK
Target 1 of the GSPC was, ‘A widely accessible working list of known plant species, as a step towards a complete world flora’, prior to revision at COP10 in Nagoya in
October 2010. Good progress has been made towards that Target with The Plant List launched at the end of
2010, pulling together available resources to produce a
‘working list’ of known species. One of the major reasons for this success was that organisations such as
Kew and Missouri existed with business aims which closely matched the Target. GSPC Targets where facilitating organisations with business aims relevant to the Target were not identified, were generally less successful. Perhaps the greatest criticism of work done at the global level on Target 1 was the relatively weak integration between national and global level processes.
Strengthening these links will be increasingly important
25

post 2010 as we seek to improve the quality of the working list and build towards the post 2010 Target of
‘An online Flora of all known plants’. One of the barriers frequently identified is the lack of national level taxonomic capacity. However, capacity building needs to be focused on key capabilities and linked to potential outputs. What lessons can be learned from progress on
GSPC Targets to date which can help us achieve the post
2010 Targets? A key element will be to ensure that ‘the
World Flora’ and supporting activities relate to national policies and make the most of existing expertise.
Furthering the GSPC goals by building capacity for plant conservation in Latin America
Montiel, OM 1
1 Missouri Botanical Garden, St Louis, USA
The Global Strategy for Plant Conservation, approved by the Conference of the Parties to the Convention on
Biological Diversity in 2002, called for building capacity for the conservation of plant diversity. The Strategy stressed the need for development of cadres of trained biologists and conservation practitioners with access to adequate resources and facilities who would be equipped to address the challenges of understanding and conserving plants. The Missouri Botanical Garden has incorporated the Strategy into its programs by engaging in capacity building through tiered training designed for people at different levels of educational background, including professional botanists and conservationists, university students, and representatives of rural communities. Together, these various programs aim to provide all levels of formal and informal training. The
Global Strategy’s recently revised and approved targets for 2020 call once more for capacity building and public engagement in accordance with national needs. The
Missouri Botanical Garden will continue to work with partners towards the fulfillment of these new goals. A particular challenge for implementation of the Strategy in
Latin America will be its incorporation into national plans and initiatives and the development of national targets to guide these efforts. We are committed to encouraging and supporting these plans and initiatives and to continuing to implement training programs that respond to national needs.
Assessing conservation status of crop genetic diversity:
Oxalis tuberosa
as a test case for new methodologies for clonally propagated crops
Emshwiller, E 1 , Epperson, BK 2
Medina, T 4 , Girón, RC 5
Catacora, P 8 , Tineo, J 9
, Nina, V
, Cruz, W 3
, Theim, T 1
6
, Tay, D 3
, Ángeles, JE 7
, Vivanco, K 3
,
, Pérez, I 10
,
1
2
University of Wisconsin – Madison, Madison, USA;
3
Michigan State University, East Lansing, USA;
4 Instituto International Potato Center, La Molina, USA;
Nacional de Innovación Agraria, Sede Central, La
Molina, USA; 5 Instituto Nacional de Innovación Agraria,
EE Santa Ana; 6 Instituto Nacional de Innovación
Agraria, EE Andenes, Peru; 7 Instituto Nacional de
Innovación Agraria, EE Baños del Inca, Peru; 8 Instituto
9
Nacional de Innovación Agraria, EE Illpa, Peru;
Instituto Nacional de Innovación Agraria, EE Canaan,
Peru; 10 Universidad Nacional de San Antonio Abad del
Cusco, Peru
One target of the Global Strategy for Plant Conservation involves conserving genetic diversity of crops. In addition, the UN-FAO's 2nd State of the World's Plant
Genetic Resources for Food and Agriculture
(SoWPGRFA) report recommends that more attention be given to 'minor crops,' recognizing that even those crops that are not among the most important ones worldwide are nonetheless crucial components of the food systems of particular world regions. The ex-situ conservation of vegetatively-propagated crops faces particular challenges compared to seed-propagated crops that can be maintained as frozen seed. Thus, complementary in-situ conservation strategies are even more needed to maintain local diversity as a vital resource for food security in rural communities. One goal of the SoWPGRFA report is to 'develop better indicators and methodologies to assess conservation status and threats.' Because conservation of crop diversity requires information about how that diversity is distributed geographically, the use of GIS techniques for the study of crop diversity is increasing.
However, this does not usually include the use of methods from geographical genetics and spatial statistics.
We conducted spatial statistical analyses of the Andean tuber crop 'oca' Oxalis tuberosa , as a model to study the evolution of clonally-propagated crops under human influence. Because clonal crops differ from seedpropagated crops in their conservation needs, we used oca as an example to study how human-mediated dispersal affects the genetic structure of clonal crop populations, i.e., how the exchange of planting material among farmers determines the distribution of oca's clonal genotypes in traditional Andean agriculture. Cultivated oca was sampled randomly in carefully distributed localities throughout the Peruvian Andes, and samples were subsequently cultivated in a highland experiment station of INIA (Instituto Nacional de Innovación
Agraria) for morphological and molecular analyses.
Spatial statistical analyses of AFLP data of 954 oca individuals from 38 localities were done for both individual AFLP alleles and clonal genotypes, the latter determined by both AFLP and morphological data. One finding with important conservation implications is that many oca clonal genotypes have very restricted geographic distributions; some were found in only four or fewer communities. Most of these were restricted to a particular area in Peru, whereas a few others were found in scattered, discontinuous areas. Our initial spatial autocorrelation analyses (Moran's I and join-count) yielded many very strong but unusual patterns across both spatial scales and genotypes. This indicates that there are other factors besides geographic distance which have shaped the distribution of genotypes, and we continue to investigate what these factors may be. In addition to the cultigen, wild O xalis species were also collected, including a wild, tuber-bearing taxon found in highland provinces of Lima Department. The latter taxon was included in continuing research to determine the progenitors of cultivated oca, but the results using AFLP data did not support these populations as likely progenitors of oca.
26

Mediterranean flora conservation – the case of
Calabria (S Italy)
Uzunov, D 1 , Caruso, G 1 , Gangale, C 2
1 Dept of Environmental and Crop Science, Marche
Polytechnic University, Ancona, Italy; 2 Natural History
Museum of Calabria, University of Calabria, Italy
Calabria region represents the southernmost part of
Italian Peninsula and is situated in the centre of the
Mediterranean basin. This area has a peculiar geological history as a part of the Calabrian Arc, quite different from the rest of the Apennines. Its flora counts more than
2500 species and subspecies, about 150 families and 850 genera. As a typical Mediterranean flora therophytes and hemicryptophytes are dominant and geophytes (16%) are relatively abundant. Species with Mediterranean distribution are about 55% and about 10% are endemic.
In the regional Red List 317 species are included (1 EX,
3 EW, 14 CR, 24 EN, 106 VU, 108 LR and 61 DD).
Present work summarizes the experiences for data collection and analysis of different type of rare and endangered species occurring in Calabria with particular attention to: Woodwardia radicans (L.) Sm. and Primula palinuri Petagna as relict species inhabiting very narrow ecological niches; Sarcopoterium spinosum (L.) Spach. and Lomelosia crenata (Cirillo) Greuter & Burdet subsp. pseudisetensis (Lacaita) Greuter & Burdet at the limit of their distribution; Ptilostemon gnaphaloides (Cyr.) Sojak,
Retama raetam (Forssk.) Webb & Berthel. subsp. gussonei (Webb) Greuter, Aizoanthemum hispanicum L. and Gentianella crispata (Vis.) Holub as examples of species with long disjunctions, Pinguicula crystallina
Sibth. & Sm. subsp. hirtiflora (Ten.) Strid. as a very rare and localized endemic taxon and Limonium sp. div. as a group with active speciation processes. The experience for application of the IUCN criteria at global and regional level for conservation status evaluation is summarized and critically analyzed. The area is under active transformation since ancient time and there is a trend to urbanizing coastal zone and abandon of mountain regions. The impact of this socio-economical processes on the flora conservation and the cumulative effect with other natural and anthropic factors are difficult to evaluate. Some approaches and models are discussed together with an analysis of the efficiency of the system of protected areas and Natura 2000 sites for plant species protection and as a response to the Global
Strategy for Plant Conservation recommendations. A model for integration of a dynamic approach into the classic ex situ and in situ conservation strategies is proposed.
Botanical education of the next generation to face national and global grand science challenges: simulating climate change research in the secondary classroom
, Thomsen, MA 1 , Thurston, S 2
1
Gerber, DT 1
University of Wisconsin-La Crosse, La Crosse, WI,
USA; 2 American Association for the Advancement of
Science, Washington DC, USA
In a recent report assessing the botanical capacity of the
United States, each of the ‘Recommendations to fill critical gaps in botanical capacity’ involved an educational component (1). In essence, an increase in a botanically educated work force would be required emphasizing the need for botanically-related student preparation at the K–12 and collegiate levels. A recent study (2) in Science suggested that research experience for K–12 classroom teachers improved high school student achievement in the classroom. To improve K–12 botanical education, we present a model for providing botanical research opportunities to pre-service teachers, i.e. college students in secondary (6th –12th grade) teacher preparation programs, connected with the creation of middle and high school level Science
NetLinks’ lesson plans. Based on the pre-service teachers’ botanical research experience, the free, online, vetted plans allow middle/high school classroom teachers to simulate climate change research conducted by field plant ecologists. References: (1) Kramer et al. 2010; (2)
Silverstein et al. (2009).
Adaptation to flooding and submergence – the
1 hormone story
Jackson, MB 1
School of Biological Sciences, Bristol, UK
Flooding of the soil or deeper submergence to include some or all the above-ground shoot threatens the survival of all vascular plant species. At the heart of the threat is severe interference with the ready gas exchange required for life-sustaining rates of aerobic respiration and photosynthesis. The longer plants are inundated the greater the risk of their failing. Even species that grow well and compete strongly in over-wet conditions cannot survive them unchanged indefinitely, although endurance varies with species, stage of development and environmental conditions such as temperature and turbidity of the flood water. Response-mode changes that increase the probability of survival are varied but may conveniently be split into metabolic adaptations that improve the efficiency with which scarce CO
2
and O
2
are used or into structural adaptations. The latter either improve accessibility to CO
2
and O
2
or reduce the severity of the consequences of damage to shoot systems by root dysfunction. The structural adaptations to flooding and submergence involve modifications to developmental pathways that are under the influence of the major hormones (auxins, cytokinins, gibberellins, abscisic acid and ethylene). Hormone production, accumulation and action are, therefore, an integral part of numerous adaptive responses of plants to flooding or submergence. Ethylene, in particular, is closely involved because it is a gas and thus prone to fast and substantial increases in concentration within plants as a result of slow escape rates imposed by external water. This offers a fast and direct signalling route that can give rise to prompt adaptive changes within hours or minutes. The paper reviews hormone involvement in developmental adaptations to flooding. These include adventitious root formation; lysigenous aerenchyma formation in roots and leaves; hypertrophic stem swelling; root to shoot
27

signalling of stomatal closure or leaf senescence and submergence-induced fast underwater elongation by stems or leaves. Some of these hormone-mediated phenomena have been studied in great depth and the findings hold immediate significance for agricultural productivity through molecular-based methods of selection. Others are less thoroughly understood but are increasingly amenable to modern methods of analysis and exploitation.
Waterproof plants: escape or quiescence
Voesenek, LACJ 1 , van Veen, H 1 , Vashist, D 1
1
Sasidharan, R 1
Utrecht University, The Netherlands
,
The semi-aquatic dicot Rumex palustris responds to complete submergence by upward movement of leaves
(hyponastic growth) and elongation of young petioles.
These two escape responses together can bring leaves above the water surface, thus restoring gas exchange with the atmosphere and increasing survival in flood-prone environments. So far our work suggests that these two responses are regulated via an ethylene-driven signaling network in which apoplastic acidification, expansin action and the activity of the hormones abscisic acid
(ABA) and gibberellin (GA) are important. R. acetosa lacks this escape response when submerged and is therefore less tolerant to flooding. The entire transcriptome of these two species during submergence was characterized using next generation sequencing technologies. The results will be discussed with special attention to the complement of genes regulated by submergence and the differences between the two species.
Mechanisms of submergence tolerance in lowland rice
Ismail, A 1 , Septiningsih, E 1 , Mackill, D 1
1 International Rice Research Institute, Philippines
Hydrological conditions in flood-prone rice ecosystems are harsh and unpredictable, leading to low and unstable productivity, as most modern varieties are sensitive to flooding. Major constraints include poor crop establishment, high mortality after complete submergence and low yield in partially or stagnant flooded areas. Prospects for enhancing yields in these lands are becoming evident with the recent progress in genetics and physiology of tolerance. Tolerance of anaerobic conditions during germination is important because of increasing interest in direct seeding. Traits associated with tolerance were studied and major QTLs were identified and are being fine-mapped for gene cloning and for use in breeding. Considerable progress was made in understanding tolerance during vegetative stage after cloning of the Submergence-1A (SUB1A) gene. Submergence induces SUB1A expression by ethylene, which interrupts the elongation escape strategy by limiting ethylene-induced GA-promoted elongation.
This conditional induction allows SUB1 to substantially enhance tolerance without changing the original traits of the popular varieties into which it is introduced. Rice accessions with reasonable tolerance of stagnant floods were recently identified and are being studied. An update
28 on current progress will be presented. Traits associated with tolerance of excess water stress offer considerable opportunities for developing resilient rice varieties to cope with current problems and with the progressively worsening conditions credited to climate changes.
Internal aeration development and the zonation of
1 plants in wetlands
Sorrell, B 1 , Tanner, C 2 , Sukias, J 2
Aarhus University, Denmark; 2 National Institute of
Water & Atmospheric Research, New Zealand
Permanent and indefinite survival of plants in flooded soils depends primarily on adaptations that increase the supply of oxygen to tissues in anoxic soil and water, usually associated with features such as (i) increased tissue porosity, (ii) changes in tissue permeability to gases, and (iii) changes in the topology of root and rhizome architecture. Differences in the degree of development of these features contribute not only to survival and growth, but are also responsible for species zonation in relation to hydrological gradients, as will be shown with examples of a range of species differing in flooding tolerance. Maintaining species diversity in managed wetlands therefore involves hydrological conditions suitable for a variety of plants that differ in their flooding tolerance. The shallowest areas of wetlands, in which soils are waterlogged but there is little standing water, can support many species which have root aeration adaptations but are otherwise unspecialised for aquatic life. Permanent standing water is a much greater challenge for plants, and survival here is restricted to species with special adaptations to their oxygen transport physiology such as the development of pressurized gas flows in their aerenchyma. These close linkages between flooding tolerance and species distributions are key considerations for maintaining species diversity in wetlands.
Submergence tolerance of plants: leaf gas films enhance oxygen and carbon dioxide exchange under water
Colmer, T 1 , Pedersen, O 2
1 The University of Western Australia, Australia;
2 University of Copenhagen, Denmark
Submergence can impact on terrestrial plants in wetlands and other low-lying areas. Plant survival underwater depends upon gas exchange for respiration (O
2
) and photosynthesis (CO
2
). Leaves of many wetland species
(e.g. Pragmites australis, Phalaris arundinacea, Oryza sativa, Hordeum marinum ) retain a thin surface layer of gas when under water. Leaf gas films facilitate O
CO
2
2
and
exchange with floodwaters. As examples, with gas films present: (i) O
2
uptake rates in darkness were up to
5-fold higher, and (ii) CO
2
uptake during light periods
(i.e. net photosynthesis) was enhanced up to 6-fold, as compared with when these films had been removed.
Improved O
2
entry during darkness and higher net photosynthesis during light periods, in submerged shoots, both enhanced internal O
2
movement via aerenchyma to roots in an anoxic substrate. When plants were submerged for 7 days with gas films removed, tissue

sugars declined and shoot and root growth were reduced, compared to submerged plants with intact gas films. In summary, leaf gas films enhance O
2
and CO during submergence, resulting in higher O
2 more CO
2 submergence tolerance of some plants.
2
exchange
in tissues and
for photosynthesis, and thus contribute to
Underwater CAM photosynthesis: benefits for carbon gain in some aquatic plants
Pedersen, O 1 , Colmer, TD 2
1 Freshwater Biological Laboratory, University of
Copenhagen, Denmark; 2 School of Plant Biology, The
University of Western Australia, Australia
CAM photosynthesis in terrestrial plant is thought to have evolved as a trait to conserve water and is thus particularly frequent among desert plants. CAM is also present in some aquatic plants inhabiting freshwater lakes and temporary pools. Here, CAM is unimportant as a water-conserving feature and instead it is considered of adaptive importance in carbon limited environments where CO
2
is stored as malate during the night and subsequently released during the day and fixed in the normal Calvin cycle. As a consequence of extensive underwater photosynthesis, CO
2
drops to very low concentration during the afternoon in vegetation-rich temporary pools. In contrast, CO
2
builds up to several times air-equilibrium during the night when respiration processes dominates. Under these environmental conditions, plants with CAM photosynthesis may maintain positive underwater net photosynthesis also during times when the external CO
2
concentration drops to sub air-equilibrium values. However, CAM photosynthesis may also restrict photorespiration which is often high in aquatic plants as a consequence of the slow gas diffusion in water compared to in air; CO addition, O
2
2
is only slowly replenished at the site of Rubisco. In
tends to build up in the tissue because of the lower solubility of O
2
compared to CO
2
. CAM activity in the present study was indicated by 9.7-fold higher leaf malate at dawn, compared with at dusk, and CAM activity was confirmed also as changes in titratable acidity (µmol H+ equivalents) of leaves. Leaves high in malate not only showed higher underwater net photosynthesis at low external CO
2
but also less apparent photorespiration. Suppression by CAM of apparent photorespiration was evident at a range of O
2 concentrations, including values below air equilibrium.
At high O of 2.2-fold atmospheric equilibrium, net
2 photosynthesis was reduced substantially and although it remained positive in leaves containing high malate, became negative in those low in malate. In situ measurements of internal leaf O
2
concentrations in
Isoëtes australis , an Australian aquatic CAM plant inhabiting granite rock pools, showed that late afternoon
O
2
increased to 32 kPa in the leaf lacunae while dropping to below 1 kPa during the night. It is thus suggested that
CAM in aquatic plants enables higher rates of underwater net photosynthesis over large O suppression of photorespiration.
2
and CO ranges in floodwaters, via increased CO
2
2
concentration
fixation and
Using ancient plant DNA to study prehistoric
1 agriculture
Brown, T 1
Manchester Interdisciplinary Biocentre, University of
Manchester, Manchester, UK
The beginning of agriculture around 10,000 years ago was a major transition in the human past. The cultivation of plants enabled humans to exert a measure of control over their food resources, protecting them from climatic and environmental uncertainty. As a result, populations grew rapidly, leading to stratified societies and the elaborate civilisations and world systems of the historic period. For the plant, domestication was an equally dramatic event that involved selection for alleles conferring phenotypes that are beneficial for the farmer, followed by enforced climate change as the plants were taken to new geographical regions. Recent work with modern landraces is emphasising the extent to which the genomes of crop plants have been shaped by adaptive evolution that has occurred since domestication. Some information on the effects of domestication on plant genomes can be inferred from comparisons of DNA sequences from landraces and wild plants. However, the study of modern material can only access genotypes that have survived to the present day, and hence does not account for the possibility that some, possibly many, of the original cultivated varieties did not survive and are not represented among modern landraces. It is also difficult to understand the tempo of the evolutionary changes from comparisons of crops and wild plants that diverged only 10,000 years ago. To address these problems, ancient DNA from cultivated plants is being studied. This work has been held back by the patchy survival of DNA in charred cereal grains, which are the commonest type of botanical material found in the archaeological record. Greater progress has been made with desiccated plant remains, which are much less widespread but which survive in reasonable quantities in
South America and in some arid areas of Africa and
Asia. In particular, it has been possible to use ancient
DNA from desiccated maize cobs from Brazil and the
Andes to map the trajectories followed during the prehistoric introduction of this plant into South America.
Much of the work with plant remains has relied on the traditional approaches to ancient DNA analysis, in which small regions of the genome are targeted by PCR and sequences obtained from the amplified fragments.
Recently, the possibility of obtaining much larger amounts of sequence data by ‘next generation’ methods has been explored. These methods are proving particularly well suited for desiccated plant remains, and extensive DNA sequences have been obtained from ancient cotton from South America and Africa as well as maize from South and Central America. It might therefore be possible to acquire complete genome sequences of archaeological plant remains, or at least for the ‘exome’, the component of the genome that codes for protein. RNA preserved in maize and cotton specimens has also been sequenced, enabling gene expression
29

patterns to be studied and regulatory RNA molecules to be identified.
Archaeogenomic evidence of local adaptation in barley
, Palmer, S 1 , Smith, O 1 , Rose, P 2 , Clapham, Allaby, R 1
1
A 3
University of Warwick, UK; 2 The Austrian
Archaeological Institute, Austria; Cairo Branch,
Zamalek, Sharia Ismail Muhammed, Cairo, Egypt;
3 Worcester Historic Environment and Archaeology
Service, Woodbury, University of Worcester, UK
Evidence from phylogeographic patterns suggests that many crop lineages existed in situ at geographic locations for millennia, giving the opportunity for specific local adaptation. The genetic variation associated with useful adaptations may be applicable to modern crop development. Barley grown by the ancient Nubians is interesting because of a phenotypic abnormality that gives the crop a 2-row appearance, despite archaeogenetic evidence that the crop was originally 6row. Moreover, apparently the same crop lineage was passed through five successive cultures, even through violent transitions. This evolutionary sequence is likely to be explained by drought adaptation, which was undoubtedly a source of stress in the area in which the archaeological site, Qasr Ibrim, is situated, upstream of the first cataract of the River Nile. Using next generation sequencing with archaeobotanical material we have established that large-scale genomic level change in plants of this region has occurred in a punctuated fashion illustrating the rapidity with which domesticated plants have evolved within the Holocene. We have also studied the archaeoepigenome and found fundamental differences in microRNA levels between the Nubian archaeological barley, and modern barley, which centre around the ‘Green Revolution’ gene interactions. These suggest that the archaeological barley was more primed than modern for germination, perhaps linked to the very short growing season available, but also had down regulation of mitochondria, which may provide the first clues as to how the abnormal phenotype was generated.
We are also employing a DNA capture approach to study the portion of the barley genome most likely to be involved with drought adaptation. The archaeogenomic evidence so far has turned up a number of surprising and independent connections to mutations associated with that part of the genome associated with the ‘Green
Revolution’ that is leading us to speculate about Green
Revolutions across the ancient world.
Molecular reconstruction of Holocene vegetation in
Scandinavia based on pollen and sediment ancient
DNA analysis
Matetovici, I 1 , Haile, J 2 , Fontana, SL 3 , Parducci, L 1
1 Dept of Ecology and Genetics, Evolutionary Biology
Centre, Uppsala University, Uppsala, Sweden; 2 Murdoch
University Ancient DNA Laboratory, Perth, Australia;
Centre for GeoGenetics, Natural History Museum of
3 Denmark, Copenhagen, Denmark; Dept of Palynology and Climate Dynamics, University of Göttingen,
Göttingen, Germany
30
Analysis of pollen and sedimentary ancient DNA isolated from sediment cores is a new emerging tool allowing past vegetation reconstruction complementary to traditional pollen and macrofossils analysis. We compared results from pollen-based vegetation analysis with results obtained using pollen and sedimentary DNA and we evaluated the results of DNA analysis using material from two Holocene sediments sampled in the
Scandes Mountains in central Scandinavia.
Genetic analysis of wheat landraces enables the location of the first agricultural sites in Italy to be identified
Brown, KA 1
1 Manchester Interdisciplinary Biocentre, Faculty of Life
Sciences, University of Manchester, UK
We typed five microsatellite loci in 52 landraces of
Italian emmer wheat to determine if genetic analysis of cereals can provide information relevant to the spread of agriculture. Each of the five loci was polymorphic with
43 allele combinations identified in the 52 landraces. The allele combinations fell into two groups. Group 1 comprised 27 genotypes found in 42 landraces and Group
2 comprised 15 genotypes found in 10 landraces. The landraces with Group 1 genotypes showed a strong correlation between geographical and genetic distances but those with Group 2 genotypes did not. We inferred that the Group 1 landraces might therefore retain a phylogeographical structure that reflects ancient events.
We present a phylogeographical model for the spread of agriculture that enables the point of origin of crop cultivation to be predicted by comparison between the genetic and geographical distances between landraces.
We applied this model to the Group 1 landraces by positioning 131 hypothetical points of origin around the coastline and northern border of Italy. The highest correlation coefficients between genetic and geographical distances were seen for hypothetical points of origin located on the coast of northern Puglia. We repeated the analysis with 1040 hypothetical points of origin located within the Italian peninsula. Again, the highest correlation coefficients were located in northern Puglia.
These predicted points of origin correspond with the location of the earliest agricultural sites in Italy, dated to c.6000 BC. The results show that plant genetics can be used to identify the location of early agricultural sites.
Bryophyte DNA extracted from soil for paleo- and extant community studies
Gussarova, G 1
K 2
, Bellemain, E 1 , Halvorsen, R 1
, Stenøien, HK 2 , Coissac, E 3
, Hassel,
1 National Centre for Biosystematics, NHM, University of
Oslo, Norway; 2 Section of Natural History, NTNU-
3 Trondheim, Norway; Laboratoire d’Ecologie Alpine,
Université Joseph Fourier, Grenoble, France
Bryophytes, especially mosses, are important components in a variety of ecosystems and well known as indicators of climate change. However, methodologies utilizing bryophyte ancient DNA in paleoenvironmental studies are lagging behind those for animals and vascular plants. We use parallel sequencing technologies and

DNA barcoding that enables species identification from environmental samples to infer both paleo- and extant species assemblages. To allow reliable species identification we built a DNA sequencing database
(chloroplast trn L intron) for the 500 most important arctic and boreal bryophyte species. Our sampling comprises several hundreds of permafrost soil cores collected throughout the Arctic region, dated from 10000 to several hundred thousand years before present. To recover bryophyte DNA from such samples, we designed new DNA barcoding markers that are suitable for amplification of degraded DNA. We also analyze modern soils sampled along a set of permanent vegetation transects to correlate soil DNA identifications with the above-ground species composition. We are now summarizing DNA sequencing datasets from the permafrost samples to be used for paleoenvironmental reconstructions and modeling. Data obtained from the modern soils will be used for developing a survey methodology to complement traditional plant community descriptions for monitoring bryophyte community structure and dynamics, including possible effects of climate change.
Paleoecology from the ground up: ancient DNA of fungi
Cross, H 1, 2 , Cooper, A 2
1 State Herbarium of South Australia, Adelaide,
Australia; 2 Australian Centre for Ancient DNA, Adelaide
University, Australia
Genetic information of subfossil fungi can provide crucial insight into the paleoecology of a region.
However, paleogenetic research on of fungi has lagged behind other areas of ancient DNA research due to critical limitations. One such constraint is the high risk of contamination from environmental sources of any potentially ancient material: fungi are found everywhere, so eliminating them is difficult. However, perhaps the most serious issue preventing progress in this field is the limited taxonomic information that is available for fungi.
Only an estimated 10% of fungal species have been described. Therefore, with any ancient DNA sequence of a fungus it is difficult to determine whether the differences observed correspond to chronological changes within taxa or merely the presence of a previously unknown modern species. Careful work in our laboratory on sedimentary and other material from a range of ages have led to progress in resolving some of the taxonomic impediments in research on ancient fungal
DNA, though serious challenges remain. All work was done under the strictest standards for ancient DNA with multiple negative controls, and environmental samples were taken from both the laboratory and source of the specimens, thus reducing the possibility of contamination. Despite the lack of genetic and taxonomic information on many fungi, by utilizing existing sequence databases and through careful analyses of results we have begun to gain an understanding of fungal species composition and diversity in past environments.
The impact and potential of fungal ancient DNA research for paleoecological studies will be discussed.
Leaf and wood traits and their afterlife effects on litter decomposition and fuel
Cornwell, W 1 , Cornelissen, H 1
1 Vrije Universiteit, Amsterdam, The Netherlands
Worldwide decomposition rates depend both on climate and the legacy of plant functional traits as litter quality.
To quantify the degree to which functional differentiation among species affects their litter decomposition rates, we brought together leaf trait and litter mass loss data for both wood and leaf litter. We show that: (i) the magnitude of species-driven differences is much larger than previously thought and greater than climate-driven variation; (ii) the decomposability of a species' litter is consistently correlated with that species' ecological strategy within different ecosystems globally, representing a new connection between whole plant carbon strategy and biogeochemical cycling. This connection between plant strategies and decomposability is crucial for both understanding vegetation–soil feedbacks, effects on fire, and for improving forecasts of the global carbon cycle.
Savannas, trees and fires across three continents: similarities and some interesting differences
Archibald, S 1 , Lehmann, C 2 , Hoffmann, B 3 , Bond, W 4
1 Natural Resources and The Environment, CSIR, South
Africa; 2 School for Environmental Research, Charles
Darwin University, Darwin, Australia; 3 Dept of Plant
4
Science, North Carolina State University, Raleigh, USA;
Dept of Botany, University of Cape Town, South Africa
Understanding the biogeography of the savanna biome has historically been coloured by the continental affiliations of the researchers involved, and contradictory perspectives are often apparent in the literature.
Moreover there is ongoing debate on the extent to which disturbance-driven feedbacks are essential for the maintenance of savanna vegetation across its environmental range. Here we report on a suite of crosscontinental analyses in Africa, Australia, and South
America which were initiated by the ARC–NZ
Vegetation Function Network. We demonstrate that much of the perceived divergence in the environmental limits of savanna between continents can be reconciled with reference to drivers of canopy closure and disturbance frequency, but that intriguing differences still remain. In particular, there is divergence between continents in the extent of arid savanna which points to a lack of understanding of the factors maintaining closedcanopy formations in arid systems. When environmental factors can not explain continental differences then phylogenetic and historical factors might be at play. We present data exploring how differences in tree allometric relationships between continents might impact rainfallbiomass relationships – with feedbacks to fire spread.
31

Xylem functional traits: coordination and scaling among hydraulic traits of plants worldwide
Jansen, S 1 , Brodribb, T
Westoby, M 3 , Choat, B 5
2 , Gleason, S 3 , Mitchell, P 4 ,
1 Ulm University, Germany;
Hobart, Australia;
Australia; 4
3
2
CSIRO, Australia;
University, Canberra, Australia
University of Tasmania,
Macquarie University, Sydney,
5 Australian National
In woody plants, xylem structure must balance three competing requirements, namely transport efficiency, safety from hydraulic failure and the mechanical support of the plant. Understanding the traits that govern this triad is important to identifying the drivers of xylem evolution as well as our ability to predict species’ response to future climate change scenarios. Because water loss is an unavoidable consequence of photosynthesis, the rate of water transport through the xylem must match the demand for water at the leaf surface; otherwise the plant may wilt and die. The key threat to hydraulic function is the suction of air into the xylem (cavitation) which occurs because plant water transport takes place under tension. If sufficient in number, the dysfunctional, air-filled conduits that result from cavitation can substantially reduce the rate of water transport and down regulate leaf gas exchange. Transport efficiency is chiefly a function of conduit size and density, but how does efficiency and its associated xylem features balance with cavitation resistance and mechanical support across a phylogenetically broad, global sampling of woody plants? With this in mind, we present a synthesis of literature data examining the functional diversity of xylem traits using a database of
681 woody species from across the globe. The database encompasses root and shoot level traits for angiosperms and gymnosperms from a broad range of biomes and phylogenetic affinities. Specifically, our analyses address
(a) how xylem traits known to confer drought-stress resistance or hydraulic efficiency vary across environmental gradients, (b) the links between transport efficiency and conduit dimensions, (c) the relationships between hydraulic and biomechanical traits, and (d) the co-ordination of xylem traits and branch level sufficiency of water transport to the leaves. We also apply principal components analyses to identify how variation in transport efficiency, cavitation resistance and biomechanical support is coordinated in multivariate trait space.
Plant population dynamics: how much variation yearto-year, site-to-site, and between species?
Buckley, YM 1 , Ramula, S
, Wardle, G 6
2 , Ehrlen, J 3 , Burns, JH 3 ,
1
Crone, EE 5
2
The University of Queensland, Brisbane, Australia;
3 University of Turku, Turku, Finland;
University, Stockholm, Sweden; 4
Stockholm
University of
California, Davis, USA;
Montana, USA; 6
5 University of Montana,
University of Sydney, Sydney, Australia
Explaining variation in population growth rates is fundamental to predicting population dynamics and population responses to environmental change. In this study, we used matrix population models, which link
32 birth, growth and survival to population growth rate, to examine how and why population growth rates vary within and among 50 terrestrial plant species. Population growth rates were more similar within species than among species; with phylogeny having a minimal influence on among-species variation. Most population growth rates decreased over the observation period and were negatively autocorrelated between years; that is, higher than average population growth rates tended to be followed by lower than average population growth rates.
Population growth rates varied more through time than space; this temporal variation was due mostly to variation in post-seedling survival and for a subset of species was partly explained by response to environmental factors, such as fire and herbivory. Stochastic population growth rates departed from mean matrix population growth rate for temporally autocorrelated environments. Our findings indicate that demographic data and models of closely related plant species cannot necessarily be used to make recommendations for conservation or control, and that post-seedling survival and the sequence of environmental conditions are critical for determining plant population growth rate.
What plant traits facilitate long distance dispersal?
Linder, HP 1 , Arroyo, M 2 , Jordan, G 3
1 University of Zurich, Switzerland;
Chile; 3
2 University of Chile,
University of Tasmania, Australia
Long distance colonisation has been established as the most frequent cause of trans-oceanic disjunctions in plants. Most recent research has been focussed on the environmental correlates of colonisation, these have been shown to be habitat or biome similarity, dominant wind and ocean direction, and the width of the ocean barriers.
Such extrinsic parameters may influence the overall frequency of dispersal among land areas, but cannot account for variation among major groups of plants.
Although plant traits that facilitate dispersal attracted much research attention in the past (e.g. strand floras, diaspore salt tolerance, ecto- and endozoochory), there have been no recent, phylogenetically corrected, investigations. Yet some traits should facilitate dispersal and establishment, while others can be assumed to be inimical to either or both these processes. Here we use a large dataset of over 11,000 austral plant species, from
45 clades, which include more than 200 long distance dispersal events. These clades were selected from all major groups of the seed plants. We use these clades, and the dispersal events they contain, to ask which traits are positively, which neutral, and which negatively associated with trans-oceanic colonisation events.
Potential traits include diaspore size, diaspore dispersal mode, life history traits (annuals vs. perennials) and habit
(herbaceous vs woody). The size of the dataset allows us to take phylogenetic effects into account by analysing each clade separately, and also to contrast traits of species which disperse only within continents, to those that show trans-oceanic dispersal. Being able to take into account taxon-specific traits that influence long distance colonisation may improve the predictiveness of environmental factors in accounting for colonisation direction and frequency.

Introducing PrometheusWiki: a Wiki for methods in ecological and environmental plant physiology
1
Nicotra, A 1 , McIntosh, E 1 , Sack, L 2
The Australian National University, Canberra,
Australia; 2 UCLA, USA
Science moves most rapidly when the majority of researchers use similar methods and can easily repeat and build upon each others' discoveries. There is a strong need for standardizing in ecological and environmental plant physiology. Traditionally information about protocols has been communicated either through conventional scientific peer-reviewed publications, which are slow and formal, or through word-of-mouth interactions and protocol sharing between specific labgroups, which is inefficient in what is now a global field.
PrometheusWiki seeks to build a new method of communication about methods within the research community. Its goal is to combine fast and informal wiki-based interactions with selected elements of traditional scientific publishing, to provide editorial review and a high quality resource. On the
PrometheusWiki site researchers can upload protocols
(and associated images, videos, code, etc) into given topics and post comments and commentary in a constructive and civil manner. PrometheusWiki uniquely combines the open communication of the www with the traditions of peer review under the oversight of an
Editorial Board of internationally recognized experts in the field. Together this ensures delivery of an up-to-date, high quality resource for anyone conducting experiments in the field of ecological and environmental physiology – students and experienced researchers alike.
PrometheusWiki launched in July 2010; here we introduce its guiding principles and structure and reflect on what has happened on the site in its first year.
Size matters: selection favours larger flowers in males but not females in a dioecious herb
Vaughton, G 1 , Ramsey, M 1
1 Botany, University of New England, Armidale, NSW,
Australia
Background and aims – Sexual selection theory predicts that male mating success is limited by access to females, whereas female mating success is limited by access to resources. In animal-pollinated plants, pollinators mediate male and female mating success rather than direct interactions between plants. Accordingly, for plant species with separate sexes (dioecy), males should be selected to enhance attractiveness to pollinators by investing more than females in floral traits that increase pollinator visitation. Such sexual dimorphism is expected to vary with sex ratio, and be greater in male-biased populations. We examine sexual selection in Wurmbea dioica by addressing the following questions: Is sexual dimorphism for flower size related to male sex ratio? Is female mating success limited by pollen availability?
Does flower size exhibit genetic variation? Does selection on flower size differ between male and females?
Methods – We assessed male sex ratio and male:female flower size dimorphism in 18 populations. In 11 populations, we assessed pollen limitation of female mating success by comparing seed set of hand-pollinated plants with that of naturally pollinated plants. In a population with a balanced sex ratio, we assessed broadsense heritability for flower size using clonal replicates.
We also used a selection-gradient approach and estimated phenotypic selection on flower size in males and females with pollen removal and seed production as fitness surrogates.
Key Results – Male sex ratio varied from 0.46–0.89 among populations. Male:female flower size dimorphism increased with increasing male sex ratio. Female seed set was not pollen-limited. Broad-sense heritability for male and female flower size was 0.78 and 0.82, respectively.
We detected strong directional selection for increased flower size in males, but weak stabilizing selection in females.
Conclusions – Sex ratio represents a unique context for selection on floral traits in W. dioica . Consistent with sex ratio-dependent selection, we found greater flower size dimorphism in male-biased populations than in lessbiased populations. We suggest that sexual selection for increased male flower size was stronger when males competed to attract pollinators, as would have occurred in male-biased populations. Interestingly, female mating success was not pollen limited in any of the populations, and we detected weak stabilizing selection for flower size. Such asymmetric selection is consistent with the male-biased attraction hypothesis of sexual dimorphism in flower size.
1
Influence of pollinator visitation patterns on multiple paternity in
Mimulus
and
Narcissus
Karron, JD 1 , Medrano, M 2 , Mitchell, RJ 3 , Herrera, CM
University of Wisconsin–Milwaukee, USA;
Biologica de Donana, CSIC, Spain; 3
2
2
Estacion
University of Akron,
USA
Animal-pollinated angiosperms cannot directly control gamete receipt or export. Instead, these plants depend on movements of pollinators whose foraging behaviors often fail to optimize the quantity or quality of pollen deposited on stigmas. Furthermore, the stochastic nature of pollinator visits often leads to striking among-flower differences in mating patterns. One factor likely to contribute to this among-flower variation is the timing and composition of sequential pollen loads deposited on each stigma. Each pollen load may differ markedly in sire representation, especially if successive pollinators approach a flower from distinct compass directions and carry pollen from different donors. Although pollen deposited during an initial floral visit may have a siring advantage due to priority, pollen grains deposited during later visits may also compete successfully for access to ovules. This multiple mating is likely to increase the genetic diversity in a sibship, which may influence competitive interactions among developing seeds within fruits and among seedlings in the field. We experimentally tested the hypothesis that sequential
33

pollinator visits contribute to the exceptionally high levels of mate diversity in fruits of Mimulus ringens
(Phrymaceae), a wetland perennial native to N. America.
We documented all bee visits to individual flowers, quantified resulting seed set, and determined paternity for
20 seeds per fruit. Most flowers (76%) received multiple visits, and the interval between visits was usually < 30 min. Flowers visited multiple times produced 44% more seeds than flowers visited once. All fruits were multiply sired. Flowers receiving a single visit averaged 3.12 outcross sires per fruit, indicating that single visits deposit pollen from several donors. The number of sires was even greater following three or more visits (4.92 outcross sires), demonstrating that sequential visits bring pollen from donors not represented in the initial visit.
Sequential pollinator visits may also contribute to the high levels of multiple paternity in Narcissus longispathus (Amaryllidaceae), a wild daffodil species endemic to a few mountain ranges in SE Spain. The effective mate number per flower varies widely among populations and years, but can be as high as 5.2 effective sires per fruit. Narcissus longispathus flowers are longlived, lasting 16.5 days in natural populations. We are currently experimentally testing the hypothesis that successive pollinator probes over multiple days increase mate diversity. We are also exploring whether visits by pollinators likely to have distinct pollen carryover characteristics lead to differences in mate diversity.
Reproductive functionality, paternity and pollen dispersal in restored
Banksia
populations
Krauss, Siegy 1
1 Kings Park And Botanic Garden, Perth, Australia
Implicit, but rarely assessed, in the success of ecological restoration projects is the management of genetic variation of restored populations and, critically, their offspring. Management of genetic variation is critical to achieving functional, self-sustaining restored populations that are resilient to environmental challenges. The delivery of robust pollinator services for seed set, inbreeding avoidance and genetic connectivity is vital, especially for animal pollinated outcrossing species. In a keystone Banksia , we assessed genetic variation, structure and differentiation of restored and natural populations, and their offspring. We also characterised mating systems and assigned paternity to offspring to characterise pollen flow, and genetic connectivity, within and among restored and adjacent natural populations, and assessed inbreeding depression through glasshouse growth trials. Restored populations and their offspring were genetically undifferentiated from, and displayed similar levels of genetic variation to, adjacent natural populations, indicating initial sourcing of genetically diverse local provenance seed. Mating system parameters were similar between populations, and extensive realized pollen dispersal within and between restored and adjacent natural populations highlight effective delivery of pollinator services and genetic connectivity and explain the absence of inbreeding depression in restored population offspring. Our results indicate successful genetic management of a restored population, from which we identify general principles relating to seed sourcing. These results add to our understanding of the importance of highly-mobile bird pollinators for the
34 facilitation of wide outcrossing and high multiple paternity for much of the flora of South West Australia.
Herkogamy influences mate diversity in fruits of the wild daffodil
Narcissus longispathus
1 , Karron, JD 2 , Mitchell, RJ,
1
Medrano, M 1
Herrera 1 , CM 1
, Requerey, R
Estación Biológica De Doñana, CSIC, SPAIN;
2 University of Wisconsin–Milwaukee, USA; of Akron, USA
3 University
The great majority of floral features have traditionally been interpreted as adaptations that enhance attraction and effectiveness of pollinators and/or promote outcrossing. Here we propose that one of the most ubiquitous floral traits among angiosperm plants, the spatial separation of anthers and stigmas within flowers
(herkogamy), may have another important function which is to enhance pollen donor diversity within a maternal sibship. We hypothesize that flowers with greater anther-stigma separation may be able to capture proportionally more diverse outcross pollen on their stigmas, and consequently increased opportunity for intense pollen competition and/or higher maternal
?ltering of male gametes could occur. This may lead to greater heterogeneity among pollen donors in seed siring success, and increased mate diversity within fruits. We test this hypothesis in Narcissus longispathus , a wild daffodil species with extensive variation in anther-stigma separation, both within and among populations (range =
0.5 to 10.4 mm). The effects of herkogamy on both the rate of outcrossing and the correlation of outcrossed paternity within fruits, and whether there is spatial or temporal variation in the effects of herkogamy on these mating parameters were studied. Mating system estimates were calculated separately for plants with high herkogamy (anther-stigma separation > 6mm) and plants with low herkogamy (anther-stigma separation < 6mm).
N. longispathus plants with high herkogamy had similar outcrossing rates (0.650) to plants with low herkogamy
(0.676). However the diversity of pollen donors siring seeds of high-herkogamy flowers was consistently greater than the diversity of pollen donors siring seeds of low-herkogamy flowers. This is the first study to demonstrate that naturally-occurring variation in a floral trait influences mate diversity within fruits.
Estimation of pollen dispersal curves and heterogeneity in male fecundity from genetic data in
1 plants
Austerlitz, F 1
Laboratoire d'Eco-Anthropologie et Ethnobiologie,
CNRS/MNHN/Universite Paris 7, Museum National d'Histoire Naturelle, Paris, France
The availability of highly polymorphic markers has yielded the possibility of estimating directly contemporaneous gene flow in plant species, in particular pollen flow. The methods use as an input genotypic data of mothers, potential fathers and of seeds collected on the mothers. They are either direct methods, based mostly on categorical or fractional paternity analyses, or indirect methods based on the comparison between the allelic

frequencies in the pollen cloud of the mothers (e.g.
TwoGener or Kindist). Both kind of methods aim at estimating the dispersal curve and the heterogeneity in male fecundities. We have now applied these methods on a substantial set of plant species, including several forest tree species. We have shown that pollen dispersal is leptokurtic in many cases, with many events of reproduction at short distances but also a small but nonnegligible part of long-distance dispersal events. We have also detected in several cases a heterogeneity of male fertilities that is generally connected with differences in some phenotypic traits. These results are important for instance in the context of conservation biology for threatened fragmented species, or more generally for the question of the spreading of advantageous and deleterious mutations over the landscape.
Patterns of paternity in populations of the endangered
Grevillea iaspicula
(Proteaceae)
Hoebee, S 1 , Young, A 2
1 La Trobe University, Bundoora, Australia;
Plant Industry, Canberra, Australia
2 CSIRO
Microsatellite loci were used in a source-sink approach to establishing patterns of paternity and to directly estimate pollen immigration into populations of the endangered shrub Grevillea iaspicula (Proteaceae). Open-pollinated progeny arrays from 5-8 individuals in each of four populations were used. In two populations, the spatial distribution, height of plants and the interaction between these parameters were assessed in order to identify whether they influenced the observed mating patterns.
Two philosophical approaches were adopted with regard to the analyses. The first approach restricted pollen donors to the source population in the first instance, then for any seed where paternity was unresolved, or there were no possible sires, paternity was re-assessed from all other populations. In the second approach, the complete dataset was used in the assignment of paternity without restriction. The majority of all seed (82%) were sired by plants located within the seed source populations. The number of pollen donors contributing to seed arrays was low (range: 1–12; mean = 5.35 ± 1.97) compared to the number of reproductive individuals, and there was significant variance in reproductive success among paternal plants. Pollen immigration rates ranged between
13.6% and 33.1%, reflecting moderate rates of gene flow
(Nm = 1.9 to 2.7). Results regarding the influence of spatial distribution and plant height were context dependent. Paternity in the smaller population was significantly correlated with plant height. Conversely, in the larger population inter-plant distance was more influential. However, multiple regression indicated that there was no simple relationship among the parameters that was consistent between populations and between approaches to paternity assignment. For G. iaspicula populations, factors such as low mate availability, only moderate gene flow and large variance in male fitness may result in reduced effective population size, compounding the effects of already small population sizes and leaving them more susceptible to environmental, demographic and genetic stochasticity, thereby reducing their long term viability.
The 'mutualism–parasitism continuum' in arbuscular mycorrhizal symbiosis: it's time for a re-think
Smith, FA 1 , Smith, SE 1
1 School of Agriculture, Food & Wine, University of
Adelaide, Australia
Growth responses of plants colonized by arbuscular mycorrhizal (AM) fungi range from highly positive to negative. Responses depend primarily on individual plant and fungal taxa and levels of soil nutrients, especially phosphate (P) and combined nitrogen (N), and have been equated to a ‘mutualism–parasitism continuum’.
Mutualism results from transfer of P and N to the plant in exchange for organic carbon (C). Lack of positive growth responses, or depressions, have been assumed to result from C transfer to the fungus with little or no P transfer to the plant, i.e. fungal parasitism. However, recent research has demonstrated that the amount of P delivered via the fungus, shown by supplying radioactive P to the external mycelium, can be very high not only where plants respond positively but also where they show no or negative responses. In the last two cases, the AM P uptake remains ‘hidden’ unless tracer P is used. Because there is no ‘extra P uptake’ into non-responsive plants, the direct P uptake via root hairs and epidermis must be lower than in non-mycorrhizal (NM) comparators; the underlying mechanism is unknown. At the same time, P transporter genes in root cortical cells are induced by AM colonization, again both in positively responsive and non- or negatively responsive plants. The transporters are involved in P uptake by the plants from the symbiotic apoplast, and expression of genes encoding them provides a marker for the potential operation of the AM pathway. The demonstration of significant P uptake via the AM pathway and reduction of direct uptake requires that researchers re-think the importance of AM symbiosis in plant nutrition at a range of scales from cells to ecology and agricultural production. About 80% of terrestrial plants, including major cereal and vegetable crops, are naturally AM in the field, except in extreme soil habitats where AM fungi are lacking. Thus, the NM condition is most often a (laboratory) artefact. At the cellular level, delivery of significant amounts of P to negatively responsive plants (which include some cereals) means that AM fungi cannot be regarded as parasitic, and the mechanism underlying the growth reductions may be P deficiency, rather than ‘excessive C drain’ as thought previously. Furthermore, research at the cellular and molecular genetic levels to understand and enhance P uptake by crops must take both the large contribution of the AM pathway and decreased direct uptake into account. Research directed to understanding and preventing decreased direct uptake may prove productive in increasing agricultural yield. Further at the whole plant level, ‘hidden’ AM P uptake may give advantages to non-responsive AM plants competing with
NM ones, that are not seen when plants are grown singly.
Overall, new information on integration of the two uptake pathways offers a possible explanation for evolutionary persistence of AM symbioses in plants that
35

are non-responsive in laboratory experiments. It is clear that the ‘parasitism’ end of the mutualism–parasitism continuum is less widespread than originally thought.
Accounting for multifunctionality in mycorrhizal symbioses when assessing the impacts of environmental change
Powell, JR 1,2 , Sikes, BA 3 , Rillig, MC 1
1 Freie Universitaet Berlin, Germany;
Western Sydney, Australia; 3
2 University of
University of Texas, Austin,
USA
While measuring the symbiotic effects of mycorrhizal fungi on their plant hosts and the ecosystem-level consequences of these interactions, researchers typically quantify either the net effects of one partner on another
(e.g., mycorrhizal dependency) or a functional trait associated with a symbiont (e.g., tissue P concentration).
However, benefits associated with mycorrhizal functioning may be derived, simultaneously or independently, via multiple mechanisms (such as nutrient uptake, pathogen protection, and modified soil-water relations, among others). Multifunctionality in mycorrhizal symbioses can make it more difficult to predict the effects of environmental change on the functioning of mycorrhizal communities. Effects may be perceived to be idiosyncratic but are actually due to the identity of the symbionts present in the study or changes in behaviour of the symbionts under different environmental conditions. In addition, complex experimental designs are required to sort out the effects of several factors at once (e.g., fertilization x pathogen inoculation x drought x mycorrhizal inoculation); having additional treatment levels for any factor quickly reduces the feasiblity of the study. An alternative approach is to target multiple traits, each associated with a functional pathway, and to simultaneously quantify these traits in manipulative experiments and observational studies where the primary gradient is one or more types of environmental change. Statistical approaches that allow for the testing of causative hypotheses from correlational data (e.g., structural equation modelling) can then be used to establish the relative importance of these multiple functions along environmental gradients based on the covariance of traits and estimates of fitness. Previous work suggests that traits such as fungal root colonization and soil hyphal length or higher-order taxonomic affiliation (family, order) can predict arbuscular mycorrhizal fungal functioning via different pathways for different host plant species, at least in one system (an abandoned agricultural field in Canada). Utilizing this approach will allow researchers to gauge the relative importance of functions they isolate and, perhaps more importantly, those they did not consider across different types of environmental change. An important consideration will be the choice of traits, or surrogates of traits, to be measured; these variables need to be useful predictors of functional outcomes during symbiotic plant-fungal interactions under the conditions that they will be estimated, but should also be suitable for highthroughput analysis. The mechanistic understanding of plant-fungal interactions facilitated by these approaches should enhance our ability to predict and prepare for the effects of environmental change in natural and managed ecosystems.
36
Arbuscular mycorrhizas and plant nutritive value in
1 a changing environment
Cavagnaro, T 1, 2 , Gleadow, R 1 , Miller, R 1
School of Biological Sciences, Monash University,
Australia; 2 Australian Centre for Biodiversity, Monash
University, Australia
The soil biota and the ecosystem services they provide are increasingly recognized as having an essential role in sustainable land management. Most terrestrial plant species, including the majority of crops, form arbuscular mycorrhizas (AM). These associations between plant roots, and a specialized group of soil fungi, play an important role in plant growth, nutrition and ecology.
Under elevated atmospheric CO plant demand for nutrients is typically increased, due to the so called CO
2
2
concentrations (eCO
2
)
fertilization effect. Given the role of
AM in plant nutrient acquisition, it has been hypothesised that AM will have an increasingly important role to play in meeting increased plant demand for nutrients in a high CO world. This however, is only
2 half of the story. Once plants acquire nutrients, irrespective of the mechanisms involved, they are incorporated into a wide array of primary and secondary metabolites, including nucleic acids, amino acids, defence compounds and many others. In our research, we have begun to consider the link between mycorrhizal acquisition of nutrients, and their fate(s) in plants. In this paper we first consider the impacts of eCO
2
on the formation and functioning of AM, with an emphasis on their role in plant nutrient acquisition. Second, we discuss the impacts of eCO
2
on plant allocation of resources (specifically N) to secondary metabolites involved in plant defence against herbivores. Finally, we begin to explore how these two quite different aspects of plant biology may be linked, and conclude with a number of readily testable hypotheses with a view to stimulating further work in this area. In so doing, we seek to better understand where our own areas of research can inform one another, and to encourage a more integrated approach to investigation of all aspects of plant responses to eCO
2
.
Effects of elevated atmospheric CO
2
, increased temperature and drought on symbiotic fungi of
Australian eucalypts
Anderson, I 1
Chambers, S 1
, Drigo, B 1 , Keniry, K 1 , Curlevski, N 1
, Ghannoum, O 1 , Tissue, D 1 , Cairney, J 1
,
1 Hawkesbury Institute for the Environment, University of
Western Sydney, Australia
Mycorrhizal fungi are central to forest carbon and nutrient cycles, but little is known about how they will respond to future climate change, especially in the context of Australian sclerophyll forests. Our recent research has used a combination of controlled environment glasshouse and field experimentation to investigate the interactive effects of elevated atmospheric
CO
2
, increased temperature and drought on Australian eucalypt mycorrhizal fungi. In a glasshouse experiment,
Eucalyptus saligna and E. sideroxylon seedlings were grown in field soil for a period of 5 months under subambient (280 ppm), ambient (380 ppm) and elevated

(640 ppm) atmospheric CO
2
conditions at both 26°C and
30°C. Molecular analyses were conducted on both DNA extracted from soil and from sand-filled hyphal ingrowth bags which select for ectomycorrhizal fungal mycelia.
Multivariate analyses showed a significant (P < 0.035) separation between soil fungal communities associated with the two different tree species. While there was no further separation based on CO
2 lower temperature (28 o
concentration at the
C), there was a clear separation between the soil fungal communities from the 280, 400 and 640 ppm CO
2
treatments at 34 o C. This response appeared to be plant-dependent at 280 and 400 ppm CO
2 however, all 640 ppm CO
2
;
samples clustered together regardless of tree species. In the field experiment, E. saligna trees were grown in 12 whole tree chambers for three years under controlled temperature conditions and exposed to either ambient (ca. 380 ppm) or elevated (ca.
640 ppm) atmospheric CO
2
concentrations and different watering regimes. Multivariate analyses showed that elevated CO
2
intensified the effect of drought stress by significantly altering fungal community composition.
Collectively, our recent data demonstrate that alterations to atmospheric CO
2
concentrations, temperature and drought conditions modify mycorrhizal and other soil fungal communities associated with Australian eucalypts.
We are currently investigating the knock-on effects of these changes for fungal driven soil processes given the potential for soil microorganisms to significantly influence the direction and magnitude of terrestrial ecosystem/atmosphere feedbacks that regulate global change.
Crassulacean acid metabolism as an ecological
1 adaptation: current status and future prospects
Smith, JAC 1 , Winter, K 1 , Holtum, JAM
University of Oxford, UK
1
Crassulacean acid metabolism (CAM) represents, together with C
3
and C
4
photosynthesis, one of the three major pathways of net CO
2
assimilation found in land plants. Because of its close association with waterlimited habitats, CAM photosynthesis is regarded as one of the clearest examples of the metabolic basis of an ecological adaptation in the plant kingdom. Whereas the underlying biochemical and physiological characteristics of CAM have been established for some time – comprising in essence nocturnal CO
2
fixation coupled with daytime stomatal closure – considerable research is now focused on the ecological diversity and evolutionary origins of this adaptation. Furthermore, as scientists contemplate a warmer world in which water availability will represent a major limiting factor for plant growth, there is intensified interest in understanding how CAM plants are so well adapted to survive (and show surprisingly high productivities) in stressful environments. Lessons learned from these studies are expected to be valuable in developing more stresstolerant, water-use-efficient crop plants, and possibly in the exploitation of CAM plants themselves on marginal land. The following areas of contemporary research on
CAM plants will be covered: (1) Biodiversity and systematics – in the light of new surveys, approximately
6 % of all angiosperm species are now estimated to possess CAM, approximately twice the number with C
4 photosynthesis, and many of these CAM plants are important components of threatened biomes, such as xerophytic succulent scrub and tropical montane cloud forests. (2) Evolutionary origins – in the absence of any significant fossil record, molecular-phylogenetic methods are being applied to certain key groups and have shown that CAM has arisen multiple times, even within individual families; date-calibrated phylogenies are being used to generate specific hypotheses about the palaeoecological context in which particular CAM lineages arose. (3) Ecological plasticity – the apparent physiological flexibility of CAM has been much discussed in the past and is being reappraised in the context of new experimental evidence revealing the extent to which this is developmentally or environmentally controlled, leading to a reassessment of the adaptive significance of plasticity in photosynthetic pathway expression. (4) Bioenergy potential – there is considerable interest in the potential for CAM plants as a sustainable bioenergy feedstock, exploiting the exceptional stress tolerance of high-biomass CAM plants such as agaves by making use of marginal, degraded land unable to support other major crops, thereby avoiding competition with land required for food, and offsetting the pressure for clearance of primary vegetation for biofuel crops in tropical regions of the world.
CAM and C
4
photosynthesis as separate but related evolutionary trajectories: examples from the
Portulacineae (Caryophyllales)
1
Edwards, E
Dept of Ecology and Evolutionary Biology, Brown
University, Providence, Rhode Island, USA
CAM and C
4
photosynthetic pathways have much in common: they employ a shared biochemical pathway that enables the concentration of CO
2
inside plant cells, they are both considered to be adaptations to stressful environments, and they are both arguably among the most convergent of complex traits, having each evolved multiple times in various plant lineages. Historically, however, they have largely been investigated as separate and unrelated adaptations, which has led us to mostly focus on the differences between CAM and C
4
plants, rather than their similarities. A broad look at the distribution of CAM and C evolutionary origins across a
4 very large phylogeny of angiosperms highlights that while CAM photosynthesis seems to have evolved more often and in a greater number of lineages, both CAM and
C plants show tight and overlapping clustering in certain
4 regions of the tree, suggesting that certain plant lineages are pre-conditioned to evolve both pathways.
Additionally, preliminary dating analyses suggest that the origins and subsequent diversification of many CAM and
C
4
lineages were contemporaneous in time. The late
Miocene/Pliocene is emerging as a critical moment in angiosperm evolutionary history, having witnessed many
C
4
origins in eudicots, the rise of C
4
-dominated grasslands, and the diversification of many of the world's ecologically important succulent CAM lineages. A closer
37

look at one of these lineages, the Portulacineae and relatives (Caryophyllales), reveals multiple origins of both CAM and C
4
pathways in a relatively small group of ~ 2200 species. I present preliminary data on vein spacing, quantitative measures of tissue succulence, gene
, recruitment, ecological habitat, and life history in C
3
CAM, and C
4
lineages in the Portulacineae and
Molluginaceae to begin to reconstruct the various steps in the evolution of both syndromes, and to identify any potential 'no turning back' scenarios, where a certain assemblage of precursor traits strongly favors the evolution of one syndrome over the other.
Ecological plasticity of crassulacean acid metabolism: drivers and constraints
Borland, A 1
1 Newcastle University, Newcastle Upon Tyne, UK
Approximately 7% of plant species, encompassing 33 families and 328 genera are known to possess a capacity for crassulacean acid metabolism (CAM). This taxonomic diversity is mirrored by the range of habitats favoured by CAM plants which range from semi-arid deserts, through tropical rainforests to aquatic ecosystems. It has been suggested that the ecological plasticity of CAM is driven by the extent to which nocturnal CO
2
uptake can be engaged or disengaged in response to perturbations in water and/or CO
2 availability. On the other hand, the anatomical and physiological traits that underpin the photosynthetic performance of CAM plants have the potential to constrain ecological plasticity in particular environments.
This presentation will highlight the mechanisms that underpin the photosynthetic plasticity of CAM and will discuss examples of how photosynthetic divergence between closely related species can be reconciled with the ecological constraints imposed by CAM.
CAM plasticity drives evolutionary success in tropical plants: a case study in Chilean
Puya
(Puyoideae,
Bromeliaceae)
Schulte, K
Smith, JAC
1,2
5
, Silvestro, D 3
, Crayn, DM 1
, Schmidt, M 3
, Novoa, P 6
, Winter, K 4
, Zizka, G 2,3,7
,
1
2
Australian Tropical Herbarium, Cairns, Australia;
Biodiversity and Climate Research Centre, Frankfurt/M,
Germany; 3 Dept. of Botany and Molecular Evolution,
4
Research Institute Senckenberg, Frankfurt/M, Germany;
Smithsonian Tropical Research Institute, Ancon,
5 Balboa, Republic of Panama; Dept of Plant Sciences,
University of Oxford, Oxford, UK; 6 Jardín Botánico
Nacional, Viña del Mar, Chile;
Frankfurt/M, Germany
7 Goethe University,
The Bromeliaceae are an important Neotropical plant family that has successfully conquered a great variety of tropical habitats. Within the family, crassulacean acid metabolism (CAM) has evolved at least four times independently: in Tillandsia, Hechtia , the Dyckia clade and in the clade comprising Puya and the Bromelioideae.
Within the latter, approximately 25 % of species in the genus Puya and 90 % of Bromelioideae are CAM plants.
Within Puya , a clade consisting of the seven Chilean species is sister to the rest. This clade is distributed along
38 a steep ecological gradient ranging from the coastal habitats of the Atacama desert to the Mediterranean and montane regions of central Chile, and contains both C
3 and CAM plants. This renders the Chilean Puya clade an important group to obtain insights into the evolution of
CAM within the more derived Bromeliaceae and a promising group to explore the correlation between the occurrence of C
3
/CAM plants and environmental parameters. In this study, phylogenetic relationships within the Chilean Puya clade were inferred using AFLP fingerprints obtained from 7 primer pairs yielding 984 characters. Samples were taken from populations of all seven species ( P. alpestris, P. berteroniana, P. boliviensis, P. chilensis, P. coerulea, P. gilmartiniae, P. venusta ) covering their geographical distribution. The photosynthetic pathway of the samples included in the phylogenetic analysis was determined from leaf tissue carbon-isotope ratio, delta 13 C. This value reflects the degree to which plants use the C pathway, in which the
3 primary carboxylating enzyme is ribulose–1,5– bisphosphate carboxylase–oxygenase, as compared with the CAM pathway, in which the primary carboxylating enzyme is PEPC, because of a kinetic isotope effect.
Extensive distribution data were collected from specimens from 27 herbaria and used to infer the ecological niche parameters of the species via the
WorldClim database. Phylogenetic analysis using maximum parsimony and Bayesian inference consistently revealed three well supported main clades termed the chilensis ( P. chilensis, P. boliviensis, P. gilmartiniae ), alpestris ( P. alpestris, P. berteroniana ), and coerulea groups ( P. coerulea, P. venusta ). The frequency distribution of delta 13 C values revealed an unexpectedly high proportion of plants with intermediate values between -22 and -18 parts per thousand indicating that these plants do not fully rely on one of the two photosynthetic modes but are using CAM in a more flexible way. Only a small proportion of plants exhibited delta 13 C values less negative than -18 parts per thousand indicative of predominantly CAM photosynthesis. The occurrence of plants with delta 13 either C
3
, CAM, or C
3
C values indicative of
/CAM intermediates was unevenly distributed among clades. The alpestris clade exhibited the greatest variability in the photosynthetic mode, whereas the coerulea clade was the least variable with a predominance of C
3
plants. Among the different climatic parameters, mean annual precipitation showed the strongest correlation with photosynthetic mode. The alpestris clade exhibited the widest ecological niche breadth, the highest variability in the photosynthetic mode employed and the widest geographical distribution.
The findings of this study lend support to the hypothesis that CAM plasticity is an important factor for the success of tropical CAM plants.
1
Bioenergy potential of CAM plants
Holtum, J 1
James Cook University, Douglas, Australia
The sources of most biofuel of plant origin are species developed for food production that are grown on productive, well-watered land best suited for growing food. The use of such prime arable land for the production of biofuel feedstock is unsustainable in light of the increasing demands for food production. Similarly,

it is generally economically unrealistic to grow biofuel feedstock on extremely marginal land because plants grow slowly when stressed, even those species that are well adapted to survive stresses. The most suitable land for growing biofuel crops tends to be land that is suboptimal for conventional crops but productive for biofuel species. In the tropics and sub-tropics such land is often seasonally dry, the type of habitat favoured by plants with crassulacean acid metabolism (CAM), a water-use efficient mode of photosynthesis. CAM species can exhibit high rates of biomass accumulation when grown in seasonally dry habitats under agronomic conditions.
For example, perennial CAM species in genera such as
Agave, Clusia and Opuntia can accumulate dry biomass at rates of 30+ tonnes ha -1 annum -1 when grown under rain-fed conditions. Such rates are comparable to, or better than, rates obtained with C
3
and C
4
biofuel crops under irrigated conditions. For most conventional biofuel crops, irrigation would be required to ensure commercially meaningful rates of biomass accumulation on seasonally dry land, thereby increasing the cost of production and reducing the net energy output of the crop. The CAM species with potential as biofuel feedstocks that are attracting commercial interest are principally within the genus Agave . Agaves accumulate fructans which have a multiplicity of uses other than for biofuel production, and contain low-lignin fibres which are amenable to hydrolysis or can be burnt to produce energy. Species such as A. tequilana are not new to agriculture, they have been bred and cultivated for many generations for alcohol production. Trials are underway in Australia and in Mexico to assess the suitability of A. tequilana as a feedstock. Other species also being considered as biofuel feedstocks include A. americana,
A. fourcroydes, A. sisalana and A. salmiana . Fastgrowing tropical Clusia that can switch between C and
3
CAM photosynthesis in response to changes in the availability of water also show promise. Physiological and morphological features of CAM enable high biomass production in periodically water-limited environments by allowing succulent CAM tissues to maintain, in a nutrient efficient manner, daily and seasonal carbon gain in the face of fluctuations in soil water supply.
The role of plant species traits contributing to invasiveness is context dependent and needs to be assessed in concert with other, potentially confounding factors
1
Pyšek, P 1,2 , Jarošík, V 1,2 , Chytrý, M 3 , Pergl, J 3
Institute of Botany, Academy of Sciences of The Czech
Republic; 2 Faculty of Science, Dept of Ecology, Charles
University Prague, Czech Republic; 3 Dept of Botany and
Zoology, Masaryk University Brno, Czech Republic
The major dynamic processes occurring within urban areas. i.e. the introduction of new plant species and their ability to naturalize and invade, need to be assessed in the overall context of biological invasions. Determining which biological and ecological traits predispose a species to become invasive is a fundamental question of invasion ecology and many studies have attempted to profile successful invaders, but traits affect invasiveness in concert with other factors. In the last decade, it has been recognized that the traits of a species are only part of any explanation for invasion success, and that other factors such as residence time and propagule pressure codetermine whether a species will successfully naturalize or invade in a new region. This recognition requires exploration of the role of traits by using models that analyze, at the same time, several groups of potentially confounding factors that co-determine the outcome of invasions. Because of this context dependence, the role of traits in plant invasions can only be assessed properly when the main confounding factors are controlled for.
Further, it becomes obvious that different factors are important at each stage of an invasion, such as introduction vs. naturalization or naturalization vs. invasion, with socioeconomic factors being generally important initially, and biogeographical, ecological, and evolutionary factors increasing in importance in later stages. This implies that different traits are crucial at different stages of the invasion process and that the relative importance of traits vs. other factors depends on the stage of invasion. Our recent studies that (i) took the above-mentioned confounding factors into account, (ii) addressed the role of traits in different stages of invasion process, and (iii) used data on the occurrence of the source species pool in other parts of the world, hence partly eliminating introduction biases, have shown that the success of particular species in early stages of invasion can be predicted by characteristics reflecting propagule pressure, residence time and adaptation to a wide range of climate, aquired in their native distribution ranges, whereas species’ biological traits are important for determining success at later stages, affecting which species become serious invaders. The results from our studies have potential practical implications: Species which have large native ranges and are common within these ranges are likely to have been favoured in terms of residence time, propagule pressure and climatic tolerance, and should be paid increased attention upon introductions. Biological traits that proved to have a direct effect on invasiveness should be taken into account in screening systems applied to evaluate deliberate introductions of alien plants to new regions. From a scientific point of view, future studies searching for invasive traits need to take into account confounding factors to avoid misleading conclusions.
Plant traits and extinction in urban areas: a metaanalysis of eleven cities
Williams, N 1 , Clements, S
McCarthy, M 1
Thompson, K 6
2 , Corlett, R
, McDonnell, M 4
, Vesk, PA 1 , Duncan, RP 7
3 , Hahs, A 4
, Schwartz, MW 5
,
,
1 The University of Melbourne, Australia;
Botanic Garden, Brooklyn, New York, USA;
4
2
3
Brooklyn
National
University of Singapore, Singapore; Australian
Research Centre for Urban Ecology, Royal Botanic
Gardens Melbourne, Victoria, Australia; 5
6
The University
University of Sheffield, UK;
7 of California, USA;
Australia Lincoln University
Urbanisation can cause local and sometimes global extinction of plant species. Using a plant functional trait approach may help to distinguish species that persist in
39

urban areas from those that have become extinct. We analyzed historical and recent survey data from 11 cities from across the globe (Adelaide, Auckland, Chicago,
Hong Kong, Los Angeles, Melbourne, New York, San
Diego, San Francisco, Singapore, Worcester) using hierarchical logistic regression models. Data were obtained for habitat and the following plant traits: growth form, clonal spread, dispersal mode, nutrient uptake strategy, spinescence, pollination system, photosynthetic pathway, plant height and seed mass. We then combined the results using meta-analysis to look for common and differing responses to urbanization. The proportion of native species that became locally extinct varied substantially among the 11 urban areas, ranging from less than 1% in San Diego to nearly 28% in Singapore. Five urban areas had extinction rates of less than 0.05% species per year, which made identifying clear correlates of extinction in noisy data difficult. For these cities there appear to be no consistent determinants of extinction outcomes and chance and idiosyncratic factors may overwhelm any trait-extinction patterns. Six cities
(Auckland, Chicago, Melbourne, New York, Singapore and Worcester) had extinction rates that exceeded 0.1% species per year which provided more power to detect patterns. In these cities both seed mass and height were strongly associated with extinction with small seeded, short plants consistently more likely to become extinct.
Given that plants in urban environments face a similar suite of filters that would be expected to select for certain traits the lack of strong and consistent patterns across cities suggest other factors such as initial abundance or preferential habitat loss may be more important drivers of urban plant extinction than plant traits.
The phylogeny of plant extinctions in Melbourne,
Australia
Hahs, A 1, 2 , Stajsic, V 3
1
McDonnell, M 3
, Udovicic, F 3 , Dobbs, C 3 ,
Australian Research Centre for Urban Ecology, Royal
Botanic Gardens Melbourne, Australia; 2 School of
Botany, The University of Melbourne, Australia;
3 National Herbarium of Victoria, Royal Botanic Gardens
Melbourne, Australia
Over half the world’s human population now lives in cities and towns. If we wish to maintain biodiversity within these urban areas, it is important to begin developing a stronger understanding of how the impacts of urbanisation affect the indigenous biota of these regions. Over the past 165 years, Melbourne, Australia has grown from a small village to a city of 4 million people. This development has not occurred without consequences for the indigenous plants of the area, with the native plant communities (Ecological Vegetation
Classes; EVCs) now represented by only 4 to 20% of their original extent. Associated with this loss of habitat has been the local extinction of fewer native plants that would be predicted by a species area curve. Recent research suggests that Melbourne may be carrying an extinction debt of 55% for its native plant species.
Examining the patterns of loss within the families and genera of species that are already locally extinct may provide some insight into which of the remaining plant species may be most vulnerable to eventual extinction.
This research investigates whether environmental
40 filtering is acting on the flora of Melbourne, and if so, which lineages appear to be more prone to extinction than others. In addressing these questions we will be building upon recent research examining variability in extinction probabilities of Melbourne’s plant species in different communities, and with different functional trait characteristics. We hope by creating a stronger understanding of the patterns of plant extinctions within
Melbourne we can facilitate the development of more effective management and conservation strategies, thereby ensuring a higher level of native biodiversity in
Melbourne than would otherwise be possible.
How lichen floras respond to urbanisation: a case study
Blanchon, D 1 , Elliott, C 1 , Kooperberg, R 1
1 Unitec Institute of Technology, Auckland, New Zealand
Lichens sensitive to air pollution have declined or disappeared in many urban areas, with only pollutionresistant species remaining. Conversely, reduction in air pollution, particularly sulphur dioxide, has led to lichens recolonising some cities. In addition to reducing pollution, many urban authorities are making efforts at restoring natural habitats. Lichens are usually not considered in ecological restoration, despite being an important part of ecological functioning, providing food, habitat, and in cyanobacterial lichens, nitrogen. Lichens often appear to have poor diversity in replanted areas, despite appropriate substrate tree species being present.
Auckland is a city centred on an isthmus on the North
Island of New Zealand. The urban area covers around
500 km 2 of a much larger (c. 6 000 km 2 ) region which includes agricultural and horticultural land, urban areas, parkland of relatively unmodified native forest, and basaltic volcanic cones and lava flows. Knowledge of the original vegetation cover is based on a few remaining urban fragments, relatively intact regional parks, herbarium records and historical accounts. This information is used to inform ecological restoration.
Originally it is thought there would have been areas of kauri ( Agathis australis ) and mixed broadleaf–podocarp forest, coastal broadleaf forest dominated by pohutukawa
( Metrosideros excelsa ), broadleaf lava forest and some wetlands. The original vegetation of the isthmus and surrounding areas have been modified by c. 700 years of human habitation, particularly forest clearance and replacement with non-native tree species. Many of the volcanic cones and lava flows have been quarried or are now covered in grasses and exotic trees. Lichens are a very visible component of the Auckland landscape, but there is no published information available on the original lichen flora of the Auckland area, or how lichens may have been affected by habitat loss or air pollution.
This project aims to identify reference ecosystems for urban Auckland, with a particular focus on corticolous lichens of lowland/coastal forest and saxicolous lichens on basaltic lava flows by examining and recording the lichens of relatively untouched areas in the nearby
Waitakere Ranges and lava flows such as those of
Rangitoto Island and the Otuataua stonefields. These reference ecosystems will then be compared with a range of datasets from Auckland, including four ‘BioBlitz’ studies of urban forest remnants and parks, in combination with surveys of the lichens of urban exotic

street trees, restoration plantings of native trees and rock walls to investigate the effects of urbanization on the lichen flora of Auckland. Initial results suggest that although high light/edge species are common in urban
Auckland and are able to colonise a range of planted native and exotic street trees, the lichens adapted to low light conditions and higher humidity (forest interior) are rarely found in urban settings. Most of the lichen species found on lava flows are still present in urban areas on rock walls.
Assessing exotic and invasive plant species of Pune urban area (India)
Mahajan, D 1 , Patil, A 2 , Waman, M 3
1
2
Dept of Botany, Waghire College, Saswad, Pune, India;
Dept of Botany, Dr D.Y. Patil College, Pimpri, Pune,
India; 3 Board of College and University Development,
University of Pune, India
Any change in floristic composition is bound to alter the primary productivity of plant species on spatio-temporal scale. Therefore, investigation of exotic species has become an imperative issue as invasion is considered a serious ecological and socio-economic problem in the
Indian subcontinent and also at a global level. The aim of this study was to document the composition of exotic and invasive plants of the Pune urban area (India). All habit forms including trees, shrubs, palms, herbs and climber species are taken into consideration. The field surveys revealed that the number of alien/exotic or introduced species has increased considerably in Pune urban area.
They were introduced initially for ornamental purpose.
Out of total exotic species the trees represented 31%, shrubs 32%, herbs 19%, climbers 10% and palms 8%.
These alien species were surveyed and documented with respect to their place of origin, habit, flowering and fruiting period, mode of propagation, and family along with the socio-economic and ecological relevance of the few species. Out of the total exotic species reported, the
American species (40%) seems to be dominating followed by Australia (16%). The species from China,
West Indies and Japan represent 7% each; Africa and Sri
Lanka 6% each, Burma 5%, Malaysia 4% and Iran 2%.
The proportion of invasive species was 18% and that of non-invasive species was 82%. However, occurrence of too many exotic species and their naturalization cannot be considered safe for native and endemic flora, especially for a region like Pune, which is in close vicinity to the globally considered biodiversity 'hotspot' belt i.e. Western Ghats. Although exotics are generally considered harmful, they do sometime play a beneficial role in ecological restoration, soil conservation and in generating new economic benefits. Field surveys and investigations reveals that deliberate introduction of a few exotics like Eucalyptus sp., Casuarina , Leucaena leucocephala, Prosopis juliflora has significantly catered to the demand of fuel, fodder and timber. Exotic species like Callistemon lanceolatus, Cassia sp., Acacia auriculiformis, Gliricidia sepium and Bougainvillea sp. have contributed largely to the aesthetic beauty of the region. Species like Jacaranda mimosifolia, Delonix regia , were also reported as dominant ornamental species in parks and gardens. The gymnospermous exotics like
Araucaria sp., Cycas revoluta, Cycas circinalis , Thuja compacta and Cupressus sp. grow luxuriantly without disturbing the native flora. However, some herbaceous
( Parthenium hysteriphorus, Cosmos sp., Cassia uniflora ), shrubby ( Eupatorium sp., Lantana camara ) and tree exotics like Leucaena leucocephala, Prosopis juliflora , etc., have turned harmful for mankind and its invasion and aggressiveness has caused ecological havoc to indigenous floristic composition. Xanthium indicum and Argemone mexicana have also created ecological problem. Parthenium hysteriphorus and Lantana camara were found to cause many allergic problems. These exotics have been naturalized and were distinguishable from the local flora only by their known history. Since the Pune urban area is located on the fringes of Western
Ghats, which is a biodiversity hotspot, introduction of alien plant species may become harmful to local species.
Eukaryotic algae colonizing building surfaces in the temperate zone – unexpected biodiversity and ecomorphological approach
Hofbauer, W 1 , Gärtner, G 2 , Rennebarth, T 1 , Breuer, K 1
1 Fraunhofer-Institute for Building Physics, Holzkirchen,
Germany;
Austria
2 Innsbruck University, Dept for Botany,
From the humid tropics it is known that aerophytic
Cyanoprokaryota form a substantial part in the colonization of building surfaces in urban areas whereas in temperate regions eukaryotic algae, especially
Chlorophytes, are prominent on this surfaces. At the
Fraunhofer-Institute for Building Physics, Holzkirchen
(Germany) and at the Botanical Institute of the
University of Innsbruck (Austria), primary microbial colonization of modern building surfaces was investigated and its biodiversity and taxonomy studied in cultures completed by physiological and genetic methods. Taxonomic investigations of the primary biological succession on the outer surface of buildings showed an unexpected rich biodiversity. The results listed in total more than 75 different species of algae
(Cyanoprokaryota and eukaryotic algae), apart from fungi, bacteria, lichens and animal organisms (in total more than 180 species), which are maintained mainly in cultures. About 20 algal species have been identified for the first time as components of the primary biological succession of biological crusts on buildings. Apart from the identified species, some need further investigation and some were found to be new species. Among them, for instance, a very peculiar species of Xanthophyceae has been isolated which appeared as an unknown species of the genus Excentrochloris . Within the last few years some new genera and species of eukaryotic algae have been described from various habitats. However it is surprising that even on the surface of modern buildings hitherto unknown species were found. The biodiversity of algae appearing on buildings is composed of different special adapted eco-forms. According to ecomorphological characteristics of the principal life form seven main units (groups, clusters) of aerophytic pioneer algae at building surfaces in the temperate region could be outlined. Unit 1 consists of unicellular forms.
This group may be divided into several sub-groups according to morphological features as form of cell, structure of cell wall, form and number of chloroplasts and ecological characteristics like light dependency or surface preference. Unit 2 is characterized by unicellular
41

forms enclosed within a shell. This group mainly is connected to rather moist surface conditions. Unit 3 comprises ecomorphologies with cells embedded in a gelatinous matrix. Unit 4 is represented by forms with a sarcinoid cell organisation and different drought resistance. Unit 5 treats unbranched filaments which sometimes may disintegrate to single cells. This group can be further organized according to nutrient requirements. Unit 6 treats with algae that form branched filaments. Unit 7 comprises macroscopic folious thalli.
Members of Trebouxiophyceae are predominant in several groups since this taxon shows a preference for aerophytic habitats. Many of the recorded algae are also known as phycobionts of different lichens. In a later stage of succession the presented units may also represent different vegetation communities, depending on prerequisites of substrate, microclimate (nanoclimate) and nutrient-supply.
Plants and global change: the ubiquity of plasticity
Valladares, F 1 , Nicotra, A 2 , Gianoli, E 3
1 CSIC, Madrid, Spain; 2 The Australian National
University, Canberra, Australia; 3 Universidad de la
Serena, Chile
When plants are exposed to changing environments they exhibit a range of phenotypic changes involving a plethora of physiological, developmental and reproductive traits. These changes have consequences not only for the performance of individual plants but also for biotic interactions and ecosystem functioning. Efforts of classifying plants according to their functional traits are challenged by the importance of adaptive plasticity for plant fitness because variation in a trait can be functionally as important as its mean, characteristic value
(or even more). Efforts of predicting future distribution of plants under different climatic scenarios are challenged by the capacity of plants to accommodate to a range of conditions by means of phenotypic plasticity.
Efforts of understanding trait mediated interactions are challenged by the fact that trait values can vary and thus the intensity and even the sign of the interaction can vary accordingly. Phenotypic plasticity is commonplace for sessile organism such as plants as a way to cope with changing environments. And phenotypic plasticity is equally ubiquitous in the research of plant responses to global change. But we are just beginning to grasp the role of plasticity not only in changing but also in real, complex scenarios of interacting factors and species.
Modelling exercises of the outcomes of species interactions and of species distributions under different climatic scenarios are hampered by the lack of progresses in this direction. We argue that progresses can be achieved by two complementary approaches: i) compiling available information and explore general trends that could be used both to model plant responses and to plan experiments and specific research, and ii) integrating conceptual approaches from different disciplines where plasticity is relevant (e.g. ecophysiology, evolutionary ecology, biogeography,
42 community ecology), which could enhance our understanding of the overall impact of plasticity for ecological and evolutionary processes involved in coping with global change. We discuss potential avenues for fruitful collaborative research illustrating these two approaches with representative examples.
Mechanisms of phenotypic plasticity and rapid evolution in novel and changing environments
Richards, C 1 , Schrey, A 1
1 Dept of Integrative Biology, University of South Florida,
Tampa, FL, USA
Understanding the mechanistic basis of phenotypic plasticity has long been of interest to evolutionary ecologists. While our understanding of the translation from genotype to phenotype is still in its infancy, most researchers across diverse fields can agree that the ability of an organism to express plasticity in a given trait must be mediated at the molecular level. Several well described environmental signaling pathways and regulatory processes in particular are amenable to experimental manipulation, and may lead to important predictions about phenotypic response to changing climates. Still, teasing apart the contributions of DNA sequence level polymorphisms from the ability of the same genotype to produce multiple phenotypes will remain a challenge. Invasive species may offer a unique opportunity for understanding the mechanisms underlying phenotypic plasticity given the reduced genetic variation following what is typically a population bottleneck effect of invasion. While the classic assumption is that dramatically reduced genetic variation will severely constrain evolutionary potential, a growing number of studies have found that many plant invaders are successful, and can even colonize new habitats with low DNA sequence based diversity. In fact a recent survey of 80 species of plants reported that surprisingly large losses (up to 50%) of heterozygosity compared to source populations do not necessarily translate into a loss of phenotypic variation. The Japanese Knotweed invasion of the northeastern part of the United States provides a good example. We used AFLPs to document very low or no DNA sequenced based diversity in phenotypically diverse populations of Japanese knotweed growing in roadside, beach and salt marsh habitats.
Using classic greenhouse and reciprocal transplant studies, we described high levels of plasticity and persistent differences in plasticity for most traits and fitness, as well as significant differences in phenotypic response to the three environments. We also found strong evidence of differentiation among populations. Given the low level of AFLP diversity, these studies highlight the possibility that phenotypic differences can contribute to establishing in novel environments and that these differences are heritable, despite low measurable DNA sequence based diversity. Recent studies show that heritable phenotypic differences may also arise from epigenetic effects and several authors have argued for the potential importance of epigenetic effects for studies in ecology and evolution. Epigenetic effects have been shown to play a role in response to hybridization and exposure to stressful or novel environments, which are circumstances often experienced by invasive plants.
Using methylation sensitive AFLP, we present evidence

that epigenetic diversity within and among Japanese knotweed populations is much higher than DNA sequence based diversity and suggest that these differences could indicate an epigenetic source of the phenotypic differentiation found in these as well as other invasive plants.
Rapid evolution of adaptive plasticity to future environments in an invasive plant
Sultan, SE 1 , Horgan-Kobelski, T 1
1 Biology Dept, Wesleyan University, Middletown, CT,
USA
As climate and CO
2
conditions quickly change, it is crucial to understand the potential of plant species to adapt to these changes. Rapid evolution of adaptive phenotypic plasticity can allow plants in natural populations to maintain fitness in the face of novel environmental conditions. Our recent work has demonstrated this kind of rapid evolutionary change of phenotypic expression in populations of the Asian shade annual Polygonum cespitosum from its introduced range in northeastern North America. In a series of glasshouse growth experiments, we tested genotypes sampled from the same three populations in 1994 and again in 2005 to track recent adaptive evolution of this invasive annual species. In only 11 generations of natural selection, genotypes from these P. cespitosum populations have evolved changes in physiological and morphological plasticity that are associated with increased plant fitness in high light conditions. These changes in plasticity are consistent with the very recent spread of the species into more open sites in its introduced range. It is particularly critical to know whether plants can maintain growth and fitness in the entirely novel environments that are predicted to arise through global climate and CO
2 change. Future environments within the North American range of P. cespitosum are expected to include high light, greater heat, drought stress, and elevated carbon dioxide concentration. We studied the plastic responses of P. cespitosum genotypes to these stresses in a manipulative field experiment at the University of Illinois Free Air
CO
2
Enrichment site (SoyFACE). As in the previous studies, we studied genotypes from the same three natural populations sampled in 1994 and again in 2005 to determine the effect of recent adaptive evolution on plasticity to these novel conditions. We measured physiological, leaf anatomical and life-history responses of these genotypes to two predicted future environments.
Both experimental field environments were hot, dry and in full sun, but one included the elevated CO
2
level predicted to occur by the year 2060.
Our findings reveal several potential limits to plant success in future environments, even in an evolutionarily labile species such as this invasive annual. First, our results show that North American populations of P. cespitosum have rapidly evolved to increase photosynthetic rate and delay senescence under full-sun, dry conditions. However, this adaptive plasticity was not expressed in the elevated CO
2
treatment. Second, the populations differed markedly in reproductive output in these novel, stressful, conditions, as well as in genetic potential for further adaptation. This result suggests that only certain populations of a species may possess the evolutionary potential to cope with rapid environmental changes such as the novel environments being created by global climate change.
Phenotypic plasticity across rainfall gradients: a case study using a congeneric
Erodium
species pair
Davidson, A 1,2 , Sheppard, A 2 , Nicotra, A 1
2
1 Australian National University, Canberra, Australia;
CSIRO, Canberra, Australia
Phenotypic plasticity is frequently referred to as playing an important role in plant invasions and in coping with environmental heterogeneity. It is also increasingly mentioned as one means by which plants, in particular, may cope with climate change. We assess plasticity in a model species pair: Erodium cicutarium, an invasive, and
E.crinitum, an Australian native. The species grow along a natural rainfall gradient and seed was collected from dry and wet ends of the range. A greenhouse experiment with five different water availabilities was conducted to assess plasticity across the full environmental gradient.
Over 30 different physiological, morphological, growth and phenological response traits were measured. We assess selection for different trait values across the water availability gradient and compare these with patterns of plasticity for each of the species and populations.
Evidence of both local adaption and phenotypic plasticity were discovered in response to water availability. The importance of plasticity and its adaptive value often differed between populations from different native rainfalls. In particular physiological characteristics such as water use efficiency (WUE) and stomatal conductance varied greatly in their plasticity between populations and was only sometimes adaptive. The implications of these findings for the species under climate change will be discussed.
Plastic and adaptive response to climatic gradients in
Waratahs (
Telopea
; Proteaceae)
Rymer, P 1 T , Offord, C 1 , Allen, C 1 , Weton, P 1 , Tissue,
1
D 2 , Rossetto, M 1
2
The Royal Botanic Gardens Sydney, Australia;
University of Western Sydney, Hawkesbury, Australia
Climate change is predicted to be the biggest threat to biodiversity. Current models of the impact of climate change on plant species do not adequately account for resilience, dispersal and adaptation, and are therefore misleading for biologists and managers. Investigations on common species across environmental gradients can provide insights into historical patterns and capacity to respond to future environmental change. The distribution of NSW Waratah ( Telopea speciosissima ) across a strong climatic gradient, and the availability of relevant background research, makes it an excellent system to explore the relative importance of phenotypic plasticity and adaptive evolution to changing climate. Nuclear microsatellite, morphological variation and climate modelling indicate coastal and upland populations have differentiated over evolutionary time scales with fluctuating levels of connectivity during Quaternary climatic cycles. We investigated how the seed germination, seedling establishment, and time of flowering of coastal and upland populations respond to
43

climatic variables in field, common garden, and laboratory experiments linked to RNA-sequencing.
Germination shows a plastic response with most events taking place across a range of temperatures (>80% 10–
30˚C). However, a significant differential population response to temperature cues is suggestive of weak adaptive evolution. A pattern supported by a reciprocal transplant experiment (RTE) of seed. Seedling growth and survivorship was explored in the field RTE and in glasshouses manipulating carbon dioxide concentration, temperature and water availability. Coupled with characterisation of morphological traits and cell structure the findings of this experiment provide insights into adaptive plasticity. Flowering in natural populations is separated by up to 8 weeks between the coast and uplands. In a common garden, the pattern flattens but the sequence of first flowering is retained with flowering of coastal followed by upland plants. Flowering is strongly genetically controlled requiring 10 days above 20˚C
(limited plasticity); however, weak differential adaptation is suggested among populations at either end of the environmental gradient. Utilising NGS Illumina platform, a comparative transcriptomic analysis of populations (coastal, uplands), organs (floral bud, germinated seed), and conditions (10˚C, 30˚C) was performed. Variation in gene expression levels and signatures of selection in coding sequences was revealed.
Functional annotation of transcripts was facilitated by the strong link with experiments (genotype–response– transcript). The final analysis will provide detailed information on the capacity for germination and flowering to change through phenotypic plasticity and adaptive evolution. The applied (robust predictions of population resilience to future climate scenarios) and theoretical (relative importance of plastic and adaptive response) will be discussed.
Below-ground stress communication in plants
Novoplansky, A 1 , Falik, O 1
1 Ben-Gurion University of The Negev, Israel
One of the main limitations of phenotypic plasticity is the lag between the perception of the environment and the time the products of the plastic responses are fully operational. Accordingly, selection is expected to promote responsiveness to cues that bear information regarding probable future conditions. We tested the hypothesis that plants are able to perceive cues of anticipated stress by eavesdropping on their stressed neighbors. Plants were grown so their roots shared their rooting volume with the roots of different neighbors.
Unstressed target plants rapidly closed their stomata following drought induction of their neighbors but no such communication took place when the roots of the stressed and the target plants did not share rooting volumes, demonstrating that the stress cues were communicated amongst roots rather than shoots.
Furthermore, shortly after the drought induction, more remote unstressed target plants, which did not share their rooting volume with the stressed plant also closed their stomata. The results demonstrate a novel type of communication whereby unstressed plants not only eavesdrop on their stressed neighbors but also leak warning signals which are perceived by additional unstressed plants. The results may have significant
44 implications for our perception of the functional organization of plant populations and communities.
Desertification of Antarctic terrestrial communities provokes rapid community change
Robinson, SA 1 , Wasley, J 2 , King, D 1
Ryan-Colton, E 1 , Mullany, K 1
, Turnbull, JD 1 ,
1 Institute for Conservation Biology and Environmental
Management, University of Wollongong, Wollongong,
2 Australia; Environmental Protection and Change;
Australian Antarctic Division; Dept of Sustainability,
Environment, Water, Population and Communities;
Kingston, Australia
Climate change is predicted to affect high latitudes first and most severely, and major changes in temperature, wind speed and stratospheric ozone have already been observed in Antarctica. Corresponding changes in vegetation have already been documented in maritime
Antarctica, where temperature changes have been particularly pronounced. For the remainder of the continent, vegetation changes have yet to be reported, and the Intergovernmental Panel on Climate Change therefore recommended improved monitoring and analysis of long-term data sets in 2007 (Anisimov et al.
2007). However, changes in temperature have also been less severe on the continent and, given the slow growth rates of the vegetation, expectations were that change might therefore be difficult to detect. Here we report the first results from a long term monitoring study established in 2003 to assess changes in vegetation in the
Windmill Islands region of East Antarctica. This area, close to Casey Station, is a biodiversity hotspot for
Antarctica, supporting some of the best-developed and most extensive plant communities on the continent. Our study monitors vegetation along a moisture gradient at two sites, transitioning from bryophyte to lichen domination with increasing aridity. Fine scale analysis shows that relative abundance of several key components of the ecosystem changed between 2003 and 2008.
Evidence that this is part of a long-term transition associated with changing water availability will be presented. Our results suggest that Antarctic terrestrial communities may be responding to climate change much faster than previously predicted, raising concerns for these old growth moss forests.
Vulnerability of Antarctic mosses to freezing injury with climate warming
Lenne, T 1 , Egerton, J 1
Bergstrom, D 4 , Ball, M 1
, Bryant, G 2 , Robinson, S 3 ,
2
1 Australian National University, Canberra, Australia;
RMIT, Melbourne, Australia; 3 University of
Wollongong, Wollongong, Australia;
Antarctic Division, Kingston, Australia
4 Australian

Productivity of mosses in Antarctica is restricted to a short growing season limited by freezing temperatures and the availability of liquid water. Analysis of the responses of mosses from three genera to freezing temperatures revealed interspecific differences in ice nucleation temperatures and tolerance of freeze-induced dehydration consistent with hydraulic anatomy and distribution along moisture gradients. During summer conditions, exposure to dehydrating effects of external ice is generally sufficient to limit vulnerability to intracellular freezing. Similarly, dehydration protects over-wintering mosses from intracellular freezing when ice nucleation temperatures of fully hydrated tissues remain well above ambient winter temperatures.
However, the mosses can become vulnerable to freezing injury when rehydration during unseasonal warming events is followed by rapid return to low winter temperatures, as occurred during the winter of 2009.
Thus, climate warming may extend the growing season, but paradoxically, increase potential injury from freezing, contributing to rapid change in the distribution and composition of moss vegetation.
Toasting plants: how extreme winter warming events may reverse the greening of the Arctic
Phoenix, G 1
1 University of Sheffield, UK
The Arctic is experiencing greatest warming in winter and a greater frequency of extreme climatic events, yet the impacts of winter climatic change and extreme events have been little studied. Of considerable concern are extreme winter warming events, during which temperatures increase rapidly causing snowmelt at landscape scales and subsequently exposing ecosystems to unseasonably warm temperatures. Following warming, ambient cold winter temperatures can return rapidly, exposing vegetation and soils to extreme cold due to the absence of the insulating snow layer. Here we report consistent findings from both a field simulation study and a natural event that show major damage to plant communities occurring at landscape scales resulting from extreme winter warming events. Extreme winter warming events were simulated in sub-Arctic heathland using heating lamps and soil warming cables. Warming resulted in up to 21-fold greater frequency of dead shoots and 47% less shoot growth of the dominant dwarf shrub
Empetrum hermaphroditum . This damage was strikingly similar to that observed following a real extreme winter warming event that occurred in December 2007. In the growing season following this event, dead dwarf shrub vegetation could be observed over large areas. Ground measurements revealed 16 times greater frequency of dead shoots and 87% less summer growth of E. hermaphroditum in visibly damages areas compared to neighbouring undamaged areas. Satellite-derived
Normalized Differential Vegetation Index values (a proxy for leaf area or photosynthetic capacity) confirmed the landscape extent of this damage and showed a considerable 26% reduction in NDVI over an area of
1424 km 2 . With increasing winter temperatures predicted along with a greater frequency of extreme climatic events, these findings suggest major consequences for the productivity and diversity of arctic ecosystems.
Critically, the damage from these winter events is opposite to the shrub expansion and greening of parts of the arctic currently attributed to summer warming. Given that the arctic is warming more in winter than summer, these impacts place a significant challenge in predicting the future of arctic vegetation in a warmer world.
Responses of boreal dwarf shrubs to changing snow conditions – a snow manipulation experiment
Saarinen, T 1 , Lundell, R 1 , Åström, H 1
Hänninen, H 1
, Rasmus, S 1 ,
1 University of Helsinki, Finland
Snow is known to have a major impact on vegetation in arctic and alpine ecosystems, but little is known about how snow affects plants in boreal forests, where the snowpack is uneven because of the tree canopy. The responses of two ericaceous dwarf shrubs, the evergreen
Vaccinium vitis-idaea (lingonberry) and the deciduous V. myrtillus (bilberry), to changing snow conditions were studied in a boreal coniferous in southern Finland. The following manipulations were carried out on 1 m 2 sample plots: (1) partial removal of snow to 15–20 cm; (2) addition of snow (the amount added was equal to the amount removed in the first treatment); (3) compression of snow to 20 cm; (4) formation of an artificial ice layer by adding distilled water on the plots. The manipulations were maintained through the winter and the ecophysiological and phenological responses were recorded. Additionally, the diurnal variation of CO
2 exchange in lingonberry leaves was estimated in late
March at the time of maximum snow depth. The thermal environment of the overwintering shoots differed substantially among the treatments. Both partial removal and compression of snow increased the occurrence of low-temperature extremes. No significant differences in winter damage, as assessed by the electrolyte leakage of the leaves (lingonberry) or shoot tips (bilberry), were recorded among the manipulations. However, partial removal of snow resulted in decreased Fv/Fm ratios of chlorophyll fluorescence in the leaves of lingonberry, which indicates increased winter stress under a thin layer of snow. Like in our previous studies, lingonberry retained its photosynthetic capacity through the winter.
The diurnal course of CO
2
exchange, estimated at the time of maximum snow depth, showed that net gain of
CO
2
was achieved at noon on all the manipulation plots excluding those of snow addition, where the intensity of
PAR remained low through the day. When integrated over the whole day, the subnivean photosynthesis compensated for a substantial proportion of the respiratory CO losses. On both density increase and
2 control plots, the compensation percentage was 50. The lowest percentage of compensation was calculated for the addition plots (15 %) and the highest percentages for the density increase and partial removal plots (72% and 80%, respectively). Snow melted on the partial removal plots on 14 April and on the addition plots on 25 April. The subsequent phenological development of lingonberry showed no significant differences among the treatments.
Bud burst occurred on 7 May and the new leaves completed their unfolding by 1 June. The timing of phenological events in bilberry showed small but significant differences among the treatments. Bud burst occurred two days earlier on the addition plots than on partial removal plots. Comparable differences were
45

observed in the timing of leaf unfolding and the beginning of flowering. In conclusion, the present study shows species-specific responses of boreal plants to changing snow conditions. Both the thickness and physical properties of snow, both of which are expected to change due to global warming, affect the overwintering success and photosynthetic performance of plants and, to lesser extent, the phenological development.
Australian alpine seed germination strategies
Hoyle, G 1 , Steadman, K
Nicotra, A 1
2 , Good, R 3
1 The Australian National University, Canberra,
2 Australia;
Australia; 3
, McIntosh, E 1 ,
University of Queensland, St Lucia,
Australia National Botanic Gardens,
Canberra, Australia
Successful plant reproduction via seed is closely associated with the seasonal timing of germination, influenced most significantly by temperature, soil moisture and light. Timing of germination dictates a seedling’s seasonal experience and thus has strong fitness consequences. Physiological dormancy mechanisms are one way seeds control the timing of germination, often postponing germination until a time when the emerging seedling has optimum chances of survival, despite dispersal into pseudo-favourable conditions. We investigated the germination strategies of 24 Australian tall alpine herb field species, spanning 11 families.
Further we examined the composition of the soil seed bank and the prevalence of dormancy in those species.
Here we present the evident range of germination phenologies throughout Australian alpine flora, in relation to plant and seed traits, and the potential consequences of a warmer, drier future climate on the optimisation of seedling establishment in the Alps.
Determining when seeds will germinate in the field contributes to our understanding of fundamental processes of population establishment, range expansion and geographic spread in order to estimate the resilience of plant and plant communities to future climate scenarios.
Current status and future predictions of UV radiation and climate change interactions
Bornman, JF 1
1 University of Waikato, New Zealand
A depleted stratospheric ozone layer (10–50 km above the Earth’s surface) has caused concern in the last few decades because of ozone’s important UV-filtering role.
In particular, the UV-B portion (280–315 nm) of the sun’s spectrum is most affected by small changes in stratospheric ozone and thus any decrease in this UVabsorbing layer has implications for the environment and living organisms. With the successful adoption of the
46
Montreal Protocol for phasing out production of ozonedepleting substances a catastrophic situation has been prevented where the UV Index, or sunburning UV radiation, could have tripled by 2065 at mid-northern latitudes (Newman et al. 2009). However, because of the long life-times of the ozone depleting substances, there are projected lag times for stratospheric ozone recovery, further complicated by a warming climate. Current prediction for the return to 1980 levels of annually averaged total column ozone at a global scale is between
2025 and 2040, while that for southern and northern midlatitudes is 2030–2040, and 2015–2030, respectively.
Over the poles, the prediction for the return to 1980 levels is between 2045–2060, and 2020–2035, for the
Antarctic (October) and Arctic (March), respectively
(Scientific Assessment of Ozone Depletion 2010). In parallel with the projected recovery of stratospheric ozone, UV irradiance will not necessarily follow this recovery, but will likely be modified by other climateinduced processes such as changes in cloud cover, aerosols, land-use (following, e.g., deforestation), and decreasing vegetative cover from rising desertification in certain regions. Changes in ozone also influence natural and human-induced climate processes, and this is resulting in complex feedback reactions not only on a climate scale, but also on the affected ecosystems. The challenge ahead in the endeavour to achieve a more stable climate will involve addressing potential future outcomes of the linkages between climate change and ozone. These include not only the biophysical, but also the political links between the Kyoto and Montreal
Protocols. There is increasing focus on modifications to plant systems by UV radiation and the interactions of different climate variables. This approach has widened our perspective on the responses to multiple stresses.
Cross-tolerance to different stresses in plants reflects some of the relationships among stresses that may lead to resilience or increased sensitivity. Thus terrestrial ecosystem response and functioning are being altered by the complexity of the impacts and feedback processes of climate change, and this is likely to intensify.
Consequently, the likely continuous evolving of plant acclimation and adaptation strategies, or in some cases, impaired functioning, underlines the importance of UV-B radiation effects within the framework of a changing climate. Regulatory and acclimative adjustments to changes in UV-B irradiance include accumulation of
UV-screening compounds, induction of antioxidants, stimulation of repair processes, as well as anatomical modifications.
Environmental effects of UV radiation on terrestrial ecosystems: cross-tolerance and ecological strategies
Hofmann, R 1 , Reichman, S 2
1
Campbell, B 3
, Wong, C 1 , Stilwell, S 1 ,
Faculty of Agriculture and Life Sciences, Lincoln
University, Canterbury, New Zealand; School of Civil,
Environmental and Chemical Engineering, RMIT
3
2
University, Melbourne, Australia; Plant & Food
Research, Mt Albert Research Centre, Auckland, New
Zealand
In their natural environment, plants are exposed to a wide spectrum of environmental variables. This may include peak levels of ultraviolet (UV) radiation in summer,

limited availability of water and nutrients, temperature extremes, salinity, pressure from herbivory and diseases, as well as other abiotic and biotic factors. While the effects of these variables can impose considerable stress on plants, frequently they can also contribute to crosstolerance. In this, one or more stress factor/s can render plants more resistant to another stress factor. A particular interest in our lab is the role of UV radiation in aboveground – belowground stress interactions. In many of these cases, flavonoids have been implicated as biochemical mediators of cross tolerance. This can be attributed to a wide array of functions for these secondary plant metabolites, including UV-screening, energy dissipation, antioxidant and herbivore-deterring capacities, involvement in signalling and modification of plant growth regulator functions. Here we use several examples to illustrate the role of flavonoids in the interplay of UV radiation with other environmental factors in natural and agricultural ecosystems. Our studies highlight the importance of UV radiation in preconditioning plants to better withstand drought stress.
In turn, we also show the importance of drought stress for mitigation of UV sensitivity, e.g. under elevated UV radiation levels in a long-term pasture ecosystem study in
New Zealand. Plant strategy theory implies that species investing constitutively into biochemical means of stress protection (‘stress tolerators’) are likely to invest less carbon into constitutive productivity. ‘Competitors’ are plant species with the opposite strategy, usually showing high levels of biomass productivity, but also higher stress sensitivity. Our findings indicate that flavonoid accumulation comes at a cost for productivity attributes in white clover populations grown under New Zealand outdoor conditions. We also present findings that show an inhibitory role of UV-B radiation on important belowground functions in wheat plants exposed to Fe deficiency. Similarly, Fe-deficiency resulted in higher
UV-B-induced increases of UV-absorbing compounds.
Our findings are currently being implemented in breeding programs for future cultivar development. In conclusion, understanding the relationships of UV radiation with other environmental variables will be crucial in the context of global climate change, where many of these interactions are likely to increase.
Increased exposure to UV-B radiation during early development leads to enhanced photoprotection and improved long-term performance of a model crop species – the role of ultraviolet radiation in sustainable crop production
Wargent, J 1 , Elfadly, E 2 , Moore, J 2 , Paul, N 2
1 Massey University, New Zealand; 2 Lancaster
University, UK
Ultraviolet (UV) radiation is a significant environmental factor, and the range of responses regulated by UV, specifically UV-B radiation (290–320nm), is vast.
Historically, responses to UV radiation have been studied largely due to concerns regarding stratospheric ozone depletion, and there is marked current uncertainty regarding the possible influence of global climate change on local UV environments throughout this century.
Despite the fact that UV-B responses have typically been considered to be damaging in nature, there are now good indications that UV-B may act as a valuable tool to enable exploitation of typical UV-B responses, leading to beneficial changes in crop quality and yield. We have evaluated such consequential changes in several crop production systems, with subsequent increases in yield observed in leafy vegetable crops following early-stage exposure to UV-B, and a variety of casual mechanisms are considered. The knock-on effects of manipulating the
UV environment for plant pathogens, herbivorous pests, and other biotic and abiotic components which comprise the majority of agroecosystems must also be considered when appraising the effects of UV-B exposure within the context of crop production. Increasing our understanding of those inherent trade-offs between negative and positive outcomes of exposure to sunlight for agroecosystems as a whole can only strengthen our ability to understand and predict UV-B response in plants and related interactions in order to further consider the now growing role of UV radiation as a regulator of crop productivity.
Cell wall sunscreens: moss sends important message about protecting against elevated UV radiation
Waterman, MJ
SA 1
1, 2 , Smith, LB 1 , Keller, PA 2 , Robinsons,
1 1Institute for Conservation Biology and Environmental
2
Management, University of Wollongong, Australia;
Centre for Medicinal Chemistry, University of
Wollongong, Australia
Ceratodon purpureus is a cosmopolitan moss species that can grow in harsh environments ranging from the icy deserts of Antarctica to hot, dry and high ultraviolet (UV) radiation Australian climates. High concentrations of ultraviolet-B (280–320 nm; UVB) absorbing compounds within the cell walls of Antarctic C. purpureus have been identified as an important photoprotective mechanism against damaging UVB radiation levels due to reduced stratospheric ozone. These compounds allow this species to be highly UV tolerant even whilst desiccated. We aimed to determine 1) the specific chemical structure of the cell wall UVB absorbing compounds in C. purpureus and 2) whether their production was induced specifically by UVB radiation. Using Australian C. purpureus as a model system we show that mosses grown in low UV environments (glasshouse conditions) have reduced cell wall UVB screening compounds. Such moss samples were then exposed to screening treatments that either 1) blocked UVB only, 2) blocked UVA and UVB or 3) transmitted 90% of solar radiation (including both UVA and UVB radiation) for 12 weeks. Moss gametophyte tissues harvested during this period were freeze-dried for methanol and alkali hydrolysis extractions. These plant extracts were analysed using UV-visible spectrophotometry and analytical high pressure liquid chromatography to monitor any change in total and specific UVB absorbing compound concentrations. These experiments show that the synthesis of cell wall-bound
UVB screens in Australian C. purpureus is induced by
UV radiation and that the time taken for UV induction suggests this enhanced synthesis occurs as new cells grow. This implies that these cell wall UVB screens have a long-term protective role in this species. This role and the structural elucidation of the cell wall-bound UVB absorbing compounds that are specifically induced will be discussed.
47

A meta-analysis of plant responses to UV-B radiation exposure in polar regions
Newsham, K 1 , Robinson, S 2
1 British Antarctic Survey, Cambridge, UK; 2 Institute for
Conservation Biology, University of Wollongong,
Australia
Vegetation in polar regions is exposed to elevated springtime UV-B radiation arising from stratospheric ozone depletion, with possible consequences for plant growth and ecosystem functioning. However, conflicting data exist in the literature as to whether plants in polar regions respond to UV-B exposure or not. In order to arrive at a consensus for how plants in these regions respond to UV-B exposure, meta-analytical methods were used to analyse data from 34 field studies into the effects of UV-B radiation on Arctic and Antarctic bryophytes and angiosperms. The studies measured plant responses to decreases in UV-B radiation under screens, natural fluctuations in UV-B irradiance, or increases in
UV-B radiation applied from fluorescent UV lamps.
Exposure to UV-B radiation was found to increase the concentrations of UV-B absorbing compounds in foliage by 7% and 25% (expressed on a mass or area basis, respectively). Exposure to UV-B radiation also reduced aboveground biomass and plant height by 15% and 10%, respectively, and increased DNA damage by 90%. No effects of UV-B exposure were found on total carotenoid or chlorophyll concentrations, photosynthetic parameters, belowground biomass, total biomass, individual leaf mass, total leaf area or specific leaf area (SLA). The methodology adopted was found to influence the concentration of UV-B absorbing compounds expressed per unit of mass, with screens and natural fluctuations in
UV-B irradiance promoting significant changes in the concentrations of these pigments, but fluorescent UV lamps failing to elicit a response, raising concerns about the use of fluorescent lamps in cold environments.
Greater reductions in leaf area and SLA, and greater increases in the concentrations of carotenoids, were found in experiments based in Antarctica than in those in the Arctic. For all but one of the response parameters
(UV-B absorbing compounds expressed on an area basis), bryophytes responded in the same way to UV-B exposure as angiosperms. Regression analyses indicated that the percentage difference in UV-B dose between treatment and control plots was positively associated with the concentrations of UV-B absorbing compounds and carotenoids, and negatively so with aboveground biomass, height and leaf area. By comparison with two other meta-analyses of plant responses to UV-B radiation, we show that the vegetation of polar regions responds to UV-B exposure in a similar way to higher plant-dominated vegetation at lower latitudes. In broad terms, plants appear able to protect themselves from elevated UV-B radiation by synthesizing UV-B absorbing compounds, whilst aboveground biomass and height are reduced by UV-B exposure, and DNA damage is increased.
Drought effects on growth and physiological status of
1 trees
Veneklaas, E 1
Plant Biology, The University of Western Australia,
Australia
Suboptimal water availability affects vegetative and reproductive growth and causes significant changes in chemical and physical properties of plant biomass. These changes may make plants more or less attractive for other organisms as a source of food, habitat, or other use. In addition, drought affects plant defence mechanisms that can control pest and disease. This paper will summarize general responses of plants to drought that are likely to affect associated organisms such as fungi and insects.
Water deficits result in reduced water status in plants, which is usually expressed (relative to well-watered plants) as reduced water content, lower turgor, more negative water potentials, and higher osmotic values. The significance of different aspects of water deficit depends on the type of relationship between host plant and pathogen or pest organism. For example, fungal endophytes are embedded in the host and their water status is closely coupled with that of the host. Leaf- and stem-sucking insects are probably more affected by hydrostatic pressures. Insects that eat whole leaves or parts of leaves may not be as much affected by water status. Insects that eat through wood have been reported to prefer lower water contents. Low water availability inevitably requires plants to reduce their water use.
Plants may reduce rates of transpiration by reducing leaf area and/or controlling stomata. These responses differ in their implications for the amount of leaf biomass present and the rate of growth, but both will reduce whole-plant productivity. Changes in leaf turnover and associated leaf age structure can alter the nutritional value of the foliage.
Longer-term adjustments of plants to low water availablity include anatomical changes. Trees produce denser wood under drought and leaves become more scleromorphous, and possibly less palatable. Shorterterm and more reversible effects of drought include changes in chemical composition of tissues. Besides dehydration-induced concentration effects, chemical changes are due to changes in metabolism and transport processes. Carbohydrates can accumulate or be depleted, depending on severity and duration of stress, and sourcesink relationships. Certain organic and inorganic compounds accumulate in leaves and roots in the process of osmotic adjustment, aimed at maintaining turgor despite dehydration. Nutrient concentrations may be affected to a greater or lesser extent by drought, depending on the relative magnitude of impacts on growth and root nutrient uptake. Belowground effects of drought are much less studied and probably more variable than aboveground effects, even in terms of root growth and physiology. Moderate drought can stimulate root growth, whereas severe drought reduces it. Roots in dry soil are architecturally, structurally and chemically different from roots in moist soil. Soil fauna and microbes that consume roots or have parasitic or
48

symbiotic relationships with roots are therefore affected by changes in soil moisture. Methodological limitations as well as the huge diversity and complexity of belowground microbial and fauna communities will continue to pose challenges to our understanding of host– pathogen/pest relationships.
Environmental influences on the cyanogenesis pathway in Forage Sorghum
O'Donnell, N 1 , Blomstedt, C
1
Hamill, J 1 , Gleadow, R 1
School of Biological Sciences, Monash University, Vic,
Australia; 2 Pacific Seeds, QLD Australia
Sorghum is a C
4
1 , Stuart, P 2 , Neale, A 1 ,
plant with high water use efficiency, making it drought and heat tolerant. Forage Sorghum is used widely in the dry tropics as fodder for cattle.
However it also produces the cyanogenic glycoside, dhurrin, and when the plant tissue is disrupted hydrogen cyanide (HCN) is released. It is thought that dhurrin may be produced as a defence to herbivory. The HCN potential (HCNp) of sorghum differs greatly between varieties, and within a variety sorghum is subject to spatial and temporal regulation. The HCNp of sorghum is known to be affected by environmental factors including drought, frost and soil fertility and varieties can differ from one season to the next. Experienced farmers know that young sorghum plants, or older plants that have been subjected to drought, are often toxic and can kill cattle.
There are three key genes that catalyse the cyanogenesis biosynthetic pathway in sorghum, two cytochrome P450s
(CYP79A1 and CYP71E1) and a (UDP)glycosyltransferase (UGT85B1). However, it is not known how these genes are regulated. We have conducted numerous experiments that have looked at the accumulation of dhurrin in various parts of sorghum plants subjected to certain stresses such as water stress, high/low nitrogen and different hormone treatments. The results have shown that the dhurrin concentration is increased under water stress, high nitrogen and when treated with some hormones. We have also look at whether there is an increase in the transcript levels of the three genes in the cyanogenesis biosynthetic pathway.
When interpreting the results we have tried to determine if the increase in dhurrin concentration is due to a reduction in plant growth or increase in activity of the three biosynthetic genes, or both. These findings are relevant to the efficient production and use of forage sorghum for farmers globally.
Cadmium exposure modifies biotic and abiotic stress signaling pathways in hyperaccumulator
Thlaspi praecox the cadmium
Llugany, M 1 , Martín, S 1 , Barcelo, J 1 , Poschenrieder, C 1
1 Universidad Autónoma De Barcelona, Spain
Metal-based defense is among the most attractive hypothesis concerning the evolutionary advantages of metal hyperaccumulation in plants. Among the different modes of action of metal defenses, the metal-therapy hypothesis is based on the frequent observation that hyperaccumulators, when grown with low metal supply, usually are highly sensitive to biotic stress.
Overproduction of signaling molecules like salicylate, as observed in a Ni-hyperaccumulating Thlaspi species, has been proposed to render these plants insensitive to infection signaling. In fact, biotic and abiotic stress responses in plants share common signaling molecules that can activate unspecific and/or specific defense mechanisms. Such biotic and abiotic elicitors include, among others, microbial cell wall molecules, compounds present in herbivore oral secretions, molecules derived from the mechanically or enzymatically damaged plant cell walls, and certain heavy metals. Jasmonic (JA) and salicylic acids (SA) are well-established components of signal transduction pathways. These molecules can act synergistically or antagonistically in the response to a variety of stress factors, leading to fine-tuning of the complex defense reactions. To further test the metaltherapy hypothesis in the heavy metal hyperaccumulation phenomenon here we analyzed endogenous levels of jasmonic and salicylic acid in T. praecox , a Cd hyperaccumulator, exposed to different biotic and abiotic stressors. Hydroponically grown plants treated or not
(control) with 50 µM Cd were exposed to different stressors: snail herbivorism, mechanical wounding, and infection by biotrophic fungus Erysiphe cruciferarum .
Endogenous levels of SA and JA were analyzed in leaf extracts of stressed and unstressed plants by liquid chromatography–electrospray ionization tandem mass spectrometry system in multiple reaction monitoring mode. Leaf SA levels in T. praecox were relatively high; in the same range as those reported by others in Ni hyperaccumulator T. goesingense . Both fungal and herbivore attack enhanced SA production in control plants. Cd supply alone tended to increase leaf SA levels, while plants exposed to both Cd and fungal or snail attack showed lower SA levels. Only when exposed to a combination of mechanical wounding and Erysiphe infection the high Cd-containing leaves showed enhanced
SA levels. Leaf JA concentrations were not enhanced by
Cd supply. This is in contrasts to the results in non hyperaccumulating species reported by others. Fungal infection and mechanical wounding tended to increase
JA levels both in controls and Cd-treated T. praecox plants, while herbivore attack induced JA production only in control plants with low Cd leaf concentrations. In conclusion, high Cd leaf concentrations in hyperaccumulator T. praecox hamper biotic-stress induced SA production, while JA signaling is less affected by Cd hyperaccumulation. To our best knowledge this is the first report on the endogenous levels of the stress signaling compounds JA and SA in a
Cd hyperaccumulator exposed to different biotic and abiotic stressors. Our results support the view of a strong influence of metal hyperaccumulation on SA signaling.
Acknowledgment: Supported by the Spanish Ministry of
Science and Innovation projects BFU2007-60332/BFI &
BFU2010-14873/BFI
Characteristics of drought-induced tree mortality and the activity of woodboring insects in the northern
1 jarrah forest of Western Australia
Matusick, G 1 , Ruthrof, K 1 , Hardy, G 1
Centre of Excellence for Climate Change Woodland &
Forest Health, Murdoch University, Australia
49

A persistent and long-term downturn in annual precipitation accentuated by an historic drought is thought to be the cause of a recent mass collapse of forest canopy species on multiple, key sites in the Northern
Jarrah-dominated forest (approx. 10,500 km 2 ). The patterns of mortality were characterized using data obtained from aerial sketch-mapping, aerial photography and geospatial databases. High levels of mortality were associated with shallow soils, which were determined by distance from outcropping granite. Eucalyptus marginata
(jarrah), Corymbia calophylla (marri), Banksia grandis , and Allocasuarina fraseriana (she-oak) have been severely affected along with a host of understorey associates. The prominence of woodboring beetle larvae
(Cerambycidae) feeding in the active sapwood and cambium of affected jarrah and marri suggests the potential for epicormic re-growth and survival is low.
These observations are exceptional considering both jarrah and marri commonly prevent mass woodborer attacks even while sustaining extensive crown dieback.
Given the exponential increase in woodborer populations and the possibility of continued tree mortality, the potential collateral effects on the jarrah forest will be discussed based on examples from native woodborer populations from Australia and around the world.
Fiendish flexibility: novelty and constraint in the adaptive phenotypic plasticity of invasive plants
Rice, K 1
1 University of California – Davis, California, USA
Very early discussions on the potential importance of evolutionary processes in biological invasions included the concepts of phenotypic plasticity and suggested that a
‘general purpose genotype’ strategy may represent an important adaptive route for invasive species. Phenotypic plasticity was considered to represent a key mode of adaptation for species invading new ranges often characterized by novel and highly variable selective pressures. I will discuss some new modes of adaptive plasticity in invasive plants, in particular, the potential importance of trans-generational plasticity (also known as maternal environmental effects) as a factor promoting plant invasion into stressful or heterogeneous environments. Trans-generational plasticity represents a form of adaptive cueing and can affect traits ranging from physiology to dispersal. As a counterpoint to this discussion of the effectiveness of adaptive plastic responses in invasive plants, I will also provide examples of factors that constrain the expression of adaptive plasticity. For example, within the invasive grass,
Bromus tectorum , genetic differentiation in phenology reduces the capacity of certain ecotypes to respond in an adaptively plastic manner to shifts in precipitation regimes. I will also present results indicating a reduction in physiological plasticity within populations of the desert weed, Bromus madritensis ssp. rubens , that have rapidly evolved lower rates of conductance in response to elevated atmospheric CO
2
. I will also discuss how phenotypic canalization, associated with stressful
50 maternal environments, may reduce the capacity of the serpentine soil invader, Aegilops triuncialis , to respond to selection. Finally I will discuss how the expression of adaptive plasticity in the invasive legume Medicago polymorpha may be amplified or constrained by variation in its rhizobial symbiont.
Temperate Asian bamboos lack common characteristics of plant invaders but not the potential
1 to invade
Mack, RN 1 , Smith, MC 1
Washington State University, Pullman, USA
Bamboos play prominent roles in Asian coniferous forests: persistent colonizers of sites after disturbance, strong competitors for light and nutrients, a source of seed and vegetative growth for predators and grazers, and fuel for recurring fires. Montane coniferous forests in the
Western United States bear striking floristic and physiognomic similarity to these Asian forests, although these forests (and much of North America north of
Mexico) are devoid of native bamboos. The increasing potential for frost-tolerant Asian bamboos to escape cultivation and become naturalized and even invasive in
U.S. coniferous forests rises as their popularity grows in ornamental horticulture. This potential occurs in spite of characteristics of these grasses that do not fit the common paradigm of invasive plants: each bamboo species displays synchronous flowering, in which each flush of flowering may be separated by many decades.
Persistence and local spread is then almost totally dependent on vigorous vegetative growth. As a result, genetic diversity in the introduced range is extraordinarily low, with representatives of single clones widely dispersed. In contrast, many bamboos possess substantial phenotypic plasticity to shade, drought and low temperature, thereby allowing them to occupy a broad environmental gradient with little or no genetic variation among individuals. Unresolved is whether their infrequent opportunities for out-crossing, natural selection and adaptive evolution will prove a severe constraint as they encounter new predators, grazers and especially parasites, while becoming much more widely distributed in North America.
How does genetic variation change during biological invasion? A study of
Pinus strobus
Mandak, B 1 , Hadincova, V 1 , Mahelka, V 1 , Wildova, R 1
1 Institute of Botany, Pruhonice, Czech Republic
Many alien species show a substantial time lag between initial establishment and the appearance of strong ecological impacts. Time lags can exist for both ecological and evolutionary reasons. Basically two scenarios could be taken into account. Firstly, a time lag can result simply from the fact that a new alien species requires time to disperse into favourable habitat patches throughout the region and to build up populations capable of producing abundant offspring. Secondly, an initial population may lack evolutionary adaptations that permit explosive population growth. After some period, through genetic reorganization within a population, an evolutionary breakthrough may occur, enabling the alien

to become an invasive species. We have tested abovementioned hypothesis using North American tree
Pinus strobus invasive in the Czech Republic. We have tested the differences in population genetic composition between native and introduced populations and compared genetic diversity among invasive and non-invasive populations within the Czech Republic. The European populations of Pinus strobus are more likely to be derived from several regions in the native range. Further analysis of population structure indicate that intraspecific hybridization among genotypes from geographically distinct regions of the native distribution range occurred following colonization in the Czech Republic.
Ecological genomics of species invasions: identifying adaptation, hybridization, and genetic tradeoffs in invading lineages
Dlugosch, K 1
1 University of Arizona, Tucson, USA
Two of the most commonly identified threats to plant diversity are loss of habitat and invasion by non-native species. While there are many clear and direct impacts of habitat loss on species abundance, it is often less clear why introduced plants become problematic for native species. Invaders can seem to be competitivey superior without displaying obvious trait differences from native weedy species. One major explanatory hypothesis for invasions is that invaders have evolved along fitness tradeoffs that are common to all plants, gaining competitive advantages at the expensive of other functions not selected in their new environment (such as defenses). An alternative (though not mutually exclusive) hypothesis is that hybridization associated with species introductions generates novel genetic benefits for invaders. Together with my collaborators, my work is aimed at evaluating the contributions of both adaptation and hybridization to species invasions, using genomic approaches to augment our understanding of the sources of trait variation in plants.
In particular, we are currently using comparative genomic techniques to identify traits and tradeoffs that facilitate the establishment and superior growth of yellow starthistle ( Centaurea solstitialis ), a noxious invader of grasslands in Mediterranean climates. We are quantifying fitness levels in response to various biotic and abiotic environments, comparing expression level and genomewide sequence variation among native and invading genotypes, and mapping the genetic basis of observed evolutionary increases in growth rate. Integrating these approaches, we show evidence for tradeoffs in fitness between benign and stressful environments, identify loci putatively under selection along these tradeoffs, and discuss potential roles for admixture and cryptic hybridization in this evolutionary change.
I also report on several methodological issues for which we have released freely-available public software, including the editing and assembly of next-generation transcriptome (EST) sequence data, and the comparison of genetic variation among outbred (wild) individuals with incomplete genomic information.
The evolutionary dynamics involved during and following colonization in an invasive plant (
Silene latifolia
)
1
Wolfe, L 1
Georgia Southern University, Statesboro, USA
The perennial plant Silene latifolia has emerged as a model system for investigating the evolutionary dynamics involved in biological invasions. This species' post-glacial expansion involved both neutral and adaptive evolution of life history traits along gradients of environmental variation in its native Europe and introduced North American range. Experimental common garden experiments conducted on both continents reveal that there has been an evolutionary shift in life history traits with N. American plants expressing a more aggressive phenotype with shorter lifespans compared to populations in the native range. One explanation for invasiveness is that selection resulting from enemy escape favors a shift in investment from defense to growth and reproduction. A large-scale survey of populations across the species' European and N.
American ranges revealed that Silene has escaped a suite of specialist enemies during colonization. Interestingly, when N. American plants are grown in the native range, they perform worse than their European conspecifics indicating that adaptation to the introduced range has occurred at the expense of performance in the native range. Yet, in addition to the importance of natural selection, aspects of Silene's colonization history suggest that it is important to consider the role that genetic drift may play in the evolution of novel phenotypes.
Molecular studies reveal the presence of genetic structure in the native range and that both a bottleneck and founder event occurred during colonization. These results hint at the prospect that the non-random introduction of a preadapted phenotype was involved during this species' initial colonization of N. America.
Rapid evolution in chemical defense despite genetic constraints in the invasive plant
Melaleuca
1 quinquenervia
Franks, S 1
Fordham University, New York, USA
Introduced species are likely to show rapid evolution, but rarely have quantitative genetic analyses been used to evaluate past and predict future evolutionary changes in invasive populations. I collected seeds of known maternity of the invasive plant Melaleuca quinquenervia throughout its native range in Australia and introduced range in Florida and planted them in common garden plots in Florida with and without the presence of two biological control insects. I quantified 20 terpenes and terpenoids from plant leaves using GC/MS. Eleven of these compounds showed higher concentrations in the native compared to introduced populations, and only one compound had a higher concentration in introduced populations. This finding indicates a potential loss of defense in the new range, in keeping with predictions of the evolution of increased competitive ability (EICA) hypothesis. I focused on three compounds (1-8 cineole,
E-nerolidol, and viridiflorol) for quantitative genetic
51

analyses. All three compounds showed positive selection differentials. Genetic variances and covariances were significant and generally larger in the native than in the introduced range populations. This suggests that the species introduction resulted in the loss of quantitative genetic variation. I found that this loss of genetic variation would potentially constrain (though not prevent) evolution based on analyses of selection gradients and G-matrix geometry. Overall the results suggest that evolutionary changes in secondary compounds have occurred following introduction of this invasive species and that the direction of evolutionary change is generally consistent with predictions from
EICA and from estimates of quantitative genetic parameters.
Algal productivity in the 21st century : the impacts of environmental change
Beardall, J 1 , Young, E 2 , Raven, J 3
1 School of Biological Sciences, Monash University,
Clayton, Australia; 2 Dept of Biological Sciences,
University of Wisconsin–Milwaukee, USA; 3 Division of
Plant Sciences, University of Dundee at SCRI, Scottish
Crop Research Institute, UK
Climate and environmental change scenarios suggest various direct and indirect impacts on algae and aquatic plants. Key environmental change factors likely to affect productivity and species diversity of marine and freshwater algae include (1) effects of elevated atmospheric CO
2
; (2) effects of elevated temperatures on algal growth and physical and chemical processes in waterbodies; and (3) changing nutrient availability related to climate change influences on ocean currents and terrestrial runoff. Ocean acidification is already occurring from increasing anthropogenic CO
2
released to the atmosphere which dissolves in the ocean increasing the dissolved CO
2
concentration. This results in a much smaller relative increase in HCO carbon, but a decrease in CO
3
3
2-
and total inorganic and release of H + , reducing pH. Experiments designed to predict effects of ocean acidification on phytoplankton have varied in methodology and experimental organisms and have suggested a variety of responses. While most phytoplankton are likely to show little direct effect of elevated CO
2
and total inorganic C, some species, including some strains of the ecologically important coccolithophorids in marine systems, are likely to show significant stimulation of photosynthesis and growth. In addition to effects of the changed inorganic carbon system on algal photosynthesis, decreases in CO
3
2 affect extracellular calcification in benthic coralline red algae, and increase the energy cost of intracellular calcification, as occurs in coccolithophores, although effects may vary with species and strain. Functional calcified structures are on the surface of organisms, so are vulnerable to dissolution if the surface ocean becomes undersaturated with aragonite (affecting many calcified macroalgae) or calcite (affecting coccolithophores). The projected rises in temperature associated with elevated greenhouse gases
52 will stimulate growth of some species but in some areas may increase ocean temperatures beyond the optima for some algal species, potentially influencing species competition. More importantly, increasing global temperature will stimulate water column stratification in lakes and in tropical and mid-latitude areas of the oceans, which will exacerbate nutrient limitation in surface waters as the photic zone is isolated from nutrient-rich deeper water. Shallower mixed layers in these areas could also increase mean exposure to photosynthetically active and UV radiation, inducing greater photoinhibitory damage to phytoplankton. Physiological experiments suggest that effects of elevated CO on algae need to be
2 considered in context with photosynthetically active radiation and the supply of other limiting nutrients.
Freshwater and coastal algae will be affected by changing nutrient availability from terrestrial runoff resulting from altered precipitation patterns and increased intensity of storm events, as well as atmospheric nutrient deposition related to anthropogenic emissions. Many of these changes will induce additive stress factors upon algae, e.g. nutrient limitation can increase sensitivity to
UVB radiation. Environmental change factors are also not expected to have uniform effects on all taxa within a habitat and so will contribute to alterations in phytoplankton and benthic algal assemblage composition and diversity. Finally, there are very few studies on genetic adaptation of algae to components of environmental change: more data are needed.
Ocean acidification: calcifiers are only the tip of the iceberg
Hurd, C 1 , Roleda, M 1 , Hutchins, D 2
1
Boyd, P 3
, Fu, F 2 , Hunter, K 1 ,
University of Otago, New Zealand;
Southern California, USA; 3
2 University of
National Institute of Water and Atmospheric Research, New Zealand
The world’s oceans have absorbed up to 50% of the carbon dioxide released by human activity since the
Industrial Revolution. The result is that our oceans are becoming more acidic, a process termed ocean acidification. Research has focused on how ocean acidification will affect the ability of marine shellfish and corals to build and maintain their calcium carbonate shells and skeletons. Marine algae (seaweeds and microscopic phytoplankton) are responsible for half of global primary productivity, and require carbon dioxide for photosynthesis. Because ocean acidification alters the availability of carbon dioxide, and other dissolved carbon sources to algae, it has the potential to affect the productivity of ALL marine algae. This fact has been largely overlooked but changes to algal productivity have wide ranging implications for marine food webs and fisheries. Using targeted physiological studies we examine which algal groups, calcifying and noncalcifying, macroscopic and microscopic, will be most susceptible to ocean acidification. Results allow predictions of the future vulnerability of algal-based coastal and open ocean ecosystems to ocean acidification.

Taxonomic and ecological studies on cyanobacteria in the catchment of Te Waihora – Lake Ellesmere
Merican, F 1 , Broady, PA 1
1 University of Canterbury, New Zealand
The distribution patterns of algal periphyton in streams are controlled by environmental factors that include land use, catchment geology, nutrient supply, shading by riparian vegetation and currents associated with stream bed morphology. These patterns could be used to monitor stream health and to inform management so as to avoid potentially detrimental proliferations. A taxonomic survey is being conducted on periphytic cyanobacteria in
Kaituna River, a tributary stream of the highly eutrophic
Lake Ellesmere / Te Waihora. Selection of sites included locations above and below human settlements and agricultural areas. Descriptions have been made of all visible macroscopic growths in the form of mats, crusts, tufts and gelatinous colonies from different in-stream microhabitats. Twenty-one morphospecies have been recorded. Gelatinous colonies comprised four morphospecies of Nostoc and one of Placoma . Matforming morphotypes are dominated by three morphospecies of Phormidium , one of Oscillatoria and one of Anabaena . The remaining thirteen morphospecies from the genera Heteroleiblenia , Tapinothrix , Calothrix ,
Chlorogloea , Cyanodermatium , Pleurocapsa and
Xenococcus are major components of epilithic crusts and tufts. Distinct patterns have been revealed in the distribution of different macroscopic growths along the stream. Although growths are more extensive in the lower reaches, fewer morphospecies are present.
Increased cover of potentially toxic oscillatorialean mats are encountered downstream of intense dairy farming activity while epilithic crusts are more common upstream where the catchment consists of native vegetation in a reserve and patches of regenerating native vegetation in low intensity sheep pasture. Cultures have been established of several dominant morphospecies for more detailed morphological analysis and investigation of morphological plasticity under different conditions. It is planned to extend the study to Selwyn River in order to compare algal periphyton in this contrasting catchment of different geology and intensity of human activity.
Algae provide solutions for global energy, water, and nutrient-sustainability challenges
Young, E
B 2
1 , Graham, L
, Zulkifly, R 2
2 , Graham, J 2 , Pfleger, B
1 University of Wisconsin–Milwaukee, USA; 2
2 , Smith,
University of Wisconsin–Madison, USA
While productivity and diversity of marine and freshwater algae in natural habitats will be affected by a range of environmental change factors, algae can also offer opportunities for mitigating some of the challenges related to global energy and water quality. The major environmental problems faced by societies worldwide include (1) dwindling supplies of fossil fuels and greenhouse gas pollution associated with their continued use, (2) the economic and ecological impacts of excess eutrophication of freshwater resources and marine coastal habitats resulting in harmful algal blooms and dead zones, and (3) increasing costs of fertilizer inputs to improve agricultural production. Algal cultivation can offer low-cost solutions to all three sustainability problems: algae can produce renewable algal biofuel feedstocks and other high-value materials, algal growth in wastewaters can remediate effluents to reduce mineral nutrient content before discharge to aquatic ecosystems, and harvesting of the algae also offers recovery of nutrient resources from effluent waters. The growth of algae as a source of biofuels has been contemplated since before the 1970s, but traditional approaches to algal cultivation for biofuels production have focused on unialgal cultures in ‘bioreactors’ which present considerable challenges for culture maintenance and cell harvesting and have often used artificial lighting or ambient conditions only available at lower latitudes.
However, recent engineering life cycle comparisons indicate that algal cultivation in wastewater has the potential to offset many of the environmental burdens of algal biomass production for biofuel feedstocks over a wider range of climatic regimes. We also propose that use of natural algal assemblages, often including a range of taxa well adapted to eutrophic conditions, is a more feasible, cheaper and is thus a more sustainable approach to algal cultivation, and can be combined with growth in wastewater effluents. Our recent experimental advances include cultivation of hypereutrophic-adapted diatoms producing up to 60 µg total lipids/mL and high cell population densities (>2.5 million cells/mL). Using a sewerage effluent water medium, we have also demonstrated cultivation of a natural community of carbohydrate-rich filamentous green algae, which are easy to harvest and can sequester substantial N and P from the effluent water, drawing soluble reactive P down to below 0.005 mg/L. This filamentous green algal community also contains a particularly desirable chemical form of cellulose that we have readily purified and hydrolyzed to glucose feedstock for bacteria genetically-modified to produce specific and uniform biofuel substrates. The addition to wastewater treatment facilities of algae-based cultivation for nutrient removal and recovery offers industry opportunities to meet increasingly stringent regulatory standards for effluent waters. This combined with a mechanism for recovering costs by the sale of organic feedstocks for industrial production of biofuels and fine chemicals offers financial incentives for municipalities and benefits to the public and environment. The co-construction of revenueproducing algal feedstock production facilities could allow developing nations to build wastewater treatment plants, thereby reducing nutrient pollution in water resources world-wide.
Promising culture medium for
Chlorella
to reduce atmospheric CO
2
Aizawa, K 1 , Miyachi, S 1
1 Institute for Clean Earth, Japan
Green house gases such as CO factors which cause climate change. Biological mitigation of atmospheric CO
The rate of CO
2
8 g CO
2
/m 2
fixation for the forest is estimated at ca.
.day. In contrast, most aquatic microalgae can more efficiently utilize CO
2
2
2
and N
2
O are important
relies on photosynthesis.
(max. 60 g CO
2
/m 2 .day, [1,
2]) at high (0.5 ~ 60 %) and air level of CO
2
53

concentration [3, 4]. CO
2
fixation, wastewater treatment, and/or the production of valuable chemicals can be carried out by the production of microalgal culture. Such production does not compete with the conventional crop production and the forestry. Therefore, photosynthetic microalgal culture has a high potential for mitigating
CO
2
. However, it has been shown that the present microalgal culture eventually emit CO
Especially, the amount of CO
2
2
and N
emitted (i.e., CO
2
2
O.
debt) during the production of chemicals used for the culture media of microalgae is relatively high. Therefore, the current well-known culture media cannot be used for
CO
2
mitigation. We have tried to develop new culture media for several microalgae which are applicable to
CO
2
mitigation. At present, an improved autotrophic culture medium for freshwater Chlorella sp. which used urea at N-limiting growth conditions was found to have very low CO
2
debt. The cells such as Chlorella regularis grew well in the new medium under high CO
2
condition as in the case of conventional ones.
Dimethyl sulphide production by marine phytoplankton: implications of climate change for an important biogeochemical process
Beardall, J 1 , Larsen, S 1
1 School of Biological Sciences, Monash University,
Clayton, Australia
Phytoplankton are major producers of dimethylsulphoniopropionate (DMSP) and its product dimethyl sulphide (DMS) which is thought to play a role in sulphur cycling and modulation of climate through cloud formation and its effect on Earth's albedo. With global climate change leading to increasing sea surface temperatures and, in some cases, freshening of marine waters, it is important to understand the impacts of these parameters on DMSP production and calcification. In this paper we present an overview on DMSP production by algae, its modulation by environmental factors such as temperature and salinity and evidence for its potential role in regulating global change. We will also present some new data on DMSP production by the coccolithophorid Gephryocapsa oceanica. DMSP production rates and cellular DMSP concentration both decreased with increasing temperature, but increased with increasing salinity. Net DMSP production became less sensitive to temperature as salinity decreased. The implications of this data for the proposed role of DMSP as a regulator of global warming will be discussed.
Ecological associations of plants–fungi from Late
Paleozoic coal-ball deposits
Wu, W 1 , Labandeira, CC 1 , Wang, S-J 2 , Hilton, J 3
1 Dept of Paleobiology, Smithsonian Institution, National
Museum of Natural History, USA; 2 State Key Laboratory of Systematic and Evolutionary Botany, Institute of
Botany, Chinese Academy of Sciences, Beijing, China;
3 School of Geography, Earth and Environmental
Sciences, University of Birmingham, Edgbaston, UK
Ecological associations among plants, fungi and arthropods long have been recognized as a critical trophic step in the evolution of wetland ecosystems, despite a sparse and uninformative fossil history. Our study examines fossil fungal and their associated plants and arthropods from anatomically preserved, permineralized coal balls from three Late Paleozoic deposits. The earliest deposit is the Late Carboniferous Calhoun Coal of the Mattoon Formation, of Kasimovian–Gzhelian age from Berryville, Illinois, USA. Of Early Permian
Asselian–Sakmarian age is a coal-ball deposit of the
Taiyuan Formation, from Xiedao Village, Xishan
Coalfield, Shanxi Province, northern China. The youngest deposit is a Late Permian coal-ball deposit from the Wangjiazhai Formation, of Changhsingian age, from the Shuicheng Mining District of Guizhou Province, southern China. The relationship of the Chinese Taiyuan floras have been determined as a major, timetransgressive expression of similar plant assemblages established earlier in wetland communities of
Euramerica. The Cathaysian floras floristically diverged during the Mississippian and continued into the Early
Permian. The latest Permian floras of southern China record the persistence of key elements of lowland swamp plant communities from the Early Permian of China and the Late Carboniferous to earliest Permian of Europe and
North America. Collectively, these units represent considerable evolution during an interval of 55 million years and a position within 15º of the paleoequator. For earlier Calhoun vegetation, plant–fungus interactions occurred principally in the extinct marattialean tree-fern
Psaronius , the dominant plant constituent, and less so for the subdominant seed fern M edullosa and the sphenophyte Sphenophyllum . For Psaronius , symbiotic and parasitic fungal structures have been found endophytically, respectively, in root and rachis tissues.
Spores and vesicular and arbuscular mycorrhizae with intra- and intercellular hyphal structures frequently occur in root tissues. Fungi in rachis cells feature complete reproductive and vegetative life stages. In one endophytic association between an insect galler and its Psaronius plant host, we note fungal colonization in the vacuities among insect coprolites (fossilized fecal pellets) and galled tissue, probably indicating a diffuse, tritrophic association. By comparison, the plant–fungus–arthropod interactions in the Xiedao plant assemblage exhibit more pervasive relationships with subdominant cordaites rather than the dominant lycopsids. The Latest Permian
Shuicheng deposit display the more active relationships between the arthropods and the host plants, including the aerial and subterranean parts. The symbiotic fungi occur principally within tissues of Psaronius group, though these tree ferns were not the dominant plants at the close of the Permian.
Aneuraceae (Metzgeriales) and Tulasnelloid fungi
(Basidiomycota): model organisms for early steps in symbiotic fungus–host plant interactions
Nebel, M 1 , Garnica, S 2 , Krause, C 2 , Quandt, D 3
1 Staatliches Museum für Naturkunde, Stuttgart,
Germany; 2 Eberhard Karls Universität Tübingen, FB1
Biologie, Tübingen, Germany; 3 Rheinische Friedrich-
54

Wilhelms-Universität, Nees-Institut für Biodiversität der
Pflanzen, Bonn, Germany
Liverworts of the family Aneuraceae are suitable organisms for comparative ultrastructural analyses because they are widespread, common, and have a relatively simple morphology. The Aneuraceae show a wide range of fungus–host plant interactions including full heteromycotrophy, which is unique among liverworts. Mycorrhizal fungi from the order
Tulasnellales (Basidiomycota), which are common in orchids, are found among liverworts only in the
Aneuraceae. Molecular phylogenetic reconstructions of the Aneuraceae resolve Riccardia as sister to the remaining monophyletic genera, with Verdoornia being the sister to Aneura (incl. Cryptothallus ) and
Lobatiriccardia . Morphological characters from the gametangia and the sporophyte formation and the mycothallus development corroborate these molecular results. The following states of fungus–host plant interactions can be observed in the Aneuraceae: 1) The fungus-free stage (e.g. Riccardia amazonica, R. andina )
2) The epiphytic stage (e.g. R. multifida ) 3) The intercellular stage ( R. intercellula ) 4) The endophytic stage: 4A) – in epidermal cells (genus Riccardia ) 4B) – in epidermal and adjacent parenchymal cells (genus
Riccardia ) 4C) – in parenchyma cells (and occasionally in epidermal cells): 4Ci) – The Riccardia type (genus
Riccardia ) 4Cii) – The Aneura type (genera Aneura,
Verdoornia and Lobatiriccardia ) 4Ciii) – The exploitative mycothallus ( Aneura [ Cryptothallus ] mirabilis ) Some observations support the hypothesis that the ecological influence is decisive for the occurrence of a particular type of fungus in a host species at the population level. For example, plants of Riccardia palmata and R. latifrons that grow on logs host closely related fungi that are usually not found in Aneura pinguis and R. multifida , which grow in wet soil and also often share closely related tulasnelloids. Different colonisation patterns of the same fungus may indicate a control of the fungal penetration by the host plant. This is the case in A. pinguis and R. multifida , where the fungi showed identical or nearly identical LSU and ITS sequences, but the colonisation type is totally different and specific.
A review of the mycorrhizae in ferns and lycophytes: their potential significance in phylogenetic and ecological studies
Lehnert, M 1 , Kessler, M 2
1
2
State Museum for Natural History, Stuttgart, Germany;
Systematic Botany, University of Zurich, Switzerland
We gathered information from literature and personal surveys about the presence, type, and abundance of mycorrhizal fungi in the sporophytes of ferns and lycophytes. Records for 1009 species and 36 of the 40 recognized fern and lycophyte families show an average presence of mycorrhiza of 68%. Only mycorrhizae involving glomeromycetes (a.k.a. arbuscular mycorrhizal fungi, AMF; 75% of infected samples) and dark-septate endophytes including ascomycetes (25% of infected samples) are known from these taxa. Glomeromycetes are dominant in terrestrial ferns and change in prevalence according to the phylogenetic position of their hosts.
Phylogenetically old lineages tend to have high colonization rates of up to 100 % while some young lineages have below-average presence of glomeromycetes. The general pattern is disturbed by young clades with strong presence of glomeromycetes
(Onocleaceae) and by old lineages with low rates of colonisation (Equisetaceae, Selaginellaceae). Epiphytic lineages are either non-mycorrhizal or have switched to another type of mycorrhizal fungus (e.g., ascomycetes), irrespective if they belong to older (Hymenophyllaceae) or younger lineages (Polypodiaceae). A reassessment of ancestral state reconstructions adding the information of mycorrhiza supports the hypothesis of a synchronous evolution of the epiphytic habit in ferns, coinciding with the angiosperms’ gain of dominance of the worldwide vegetation. The information about the mycorrhizal colonisation of some groups (e.g., lycophytes,
Oleandraceae, Dennstaedtiaceae, Tectariacaeae) is still insufficient and calls for more intensive studies. Reports for gametophytes indicate a much clearer trend of successive independence of ferns and lycophytes from mycorrhizae with advancing evolution.
Insights into the multiple origins of symbioses between
Acrospermum
and polypod ferns: evidence from plant and fungal sequence data
Sundue, MA 1 , Doyle, V 2 , Ranker, TA 3
1 Pringle Herbarium, Plant Biology Dept, University of
Vermont, Burlington, USA;
Garden, Bronx, NY, USA; 3
2 The New York Botanical
Dept of Botany, University of
Hawaii, Honolulu, Hawai'i, USA
Among ferns, Polypodiaceaeare unique in having welldeveloped relationships with the bitunicate ascomycete genus Acrospermum (Dothidiomycetes +
Arthoniomycetes), a genus of about 60 spp. known from temperate and tropical America, and Europe. Most species of Acrospermum are saproprobic; however, some are epibiotic and are known to inhabit mosses and ferns.This is best developed within two clades of tropical
American grammitid ferns which correspond to the genera Ascogrammitis Sundue(17 spp.) and Mycopteris
Sundue ined. (ca. 30 spp.)These genera of ferns have a near constant relationship with Acrospermum ; the ascomes of which can be found on nearly every leaf of every plant throughout the range of both genera. One species, A. maxonii Farlow ex Riddle, has been described from the leaves of grammitid ferns, and is accepted as the species that is present upon all grammitid ferns.
However, morphometric studies suggest that above 3000 m on the eastern slope of the Andes, a second undescribed species with more orbicular ascomes is present instead of A.maxonii
. The non-grammitid polypod ferns Pleopeltis ballivianii (Rosenst.) A.R.Sm.,
P. furfuraceum (Schltdl. & Cham.) A.R.Sm., and
Campyloneurum lorentzii (Hieron.) Chingare also host to
Acrospermum , but with less consistent relationships associations, and with the range of the host-plant exceeding that of the fungus. Phylogenetic analysis of a broad sampling of Ascomycetes using [SSU, LSU] support the monophyly of Acrospermum , and the sister relationship between A. maxonii and the aforementioned undescribed high-elevation species. Phylogenetic analyses of polypod ferns using chloroplast markers
[ atp B, rbc L, trn L-F] support four independent colonization events of Acrospermum onto these ferns.
55

Results also suggest that within the grammitid clades that are associated with Acrospermum , the fungi are more closely associated with habitat than they are with particular host species.
Mycorrhizal colonizations of ferns and lycophytes on the island of La Réunion in relation to nutrient availability
Kessler, M 1 , Jonas, R 2 , Strasberg, D 3 , Lehnert, M 4
1 University of Zurich, Switzerland;
3
2 University of
Göttingen, Germany;
Réunion; 4
University of La Réunion, La
Staatliches Museum für Naturkunde, Stuttgart,
Germany
Very little is known about the degree and ecological correlates of mycorrhization of lycophytes and ferns, although mycorrhizae are believed to be crucial for the majority of land plants. We screened the degree of mycorrhizal colonization for all 75 fern species recorded in nine survey plots across a wide range of soil conditions on the island of La Réunion, including five plots on lava flows of different age. Overall, 66% of all samples had mycorrhizae, but ground-dwelling species had significantly higher colonization levels (78%) than epiphytes (58%). Among ground-dwelling species, 98% of the individuals belonged to species with mycorrhizae, which was significantly higher than at the species level.
Most samples had glomeromycetes as fungal partners, but septate endophytes were found in 41% of the colonized samples. While we found no significant relationship between the percentage of fern species with mycorrhizae per plot and seven ecological parameters, the percentage of fern individuals with mycorrhizal colonizations per plot significantly increased on shallow soils with high pH values and high base-saturation. This supports the idea that mycorrhizal colonizations confer an ecological advantage to colonized individuals, and that this advantage is more pronounced on nutrientdeficient sites. Our study thus provides evidence for an ecological advantage of mycorrhizae for ferns, but raises the question why, despite this advantage, species-level mycorrhization among ferns (68%) is so much lower than the average for land plants (85%).
Self-incompatibility and interspecific hybridization in
Senecio
1
Hiscock, SJ 1 , Brennan, AC 2
School of Biological Sciences, University of Bristol,
Bristol, UK; 2 Royal Botanic Garden Edinburgh,
Edinburgh, UK
The Senecio (ragwort) study system is a well known example of recent homoploid hybrid speciation. Two species of Senecio , S. chrysanthemifolius and S. aethnensis , are capable of producing fertile hybrids and form a natural stable hybrid zone along an altitudinal gradient on Mount Etna, Sicily. A third species, S. squalidus , is recognized as a recent homoploid hybrid
56 species derived from this hybrid zone that originated following the introduction of hybrid plants to the UK about 300 years ago. Invasive S. squalidus now occupies novel urban wasteground, railway, and motorway habitats throughout the UK. All three Senecio species have an outcrossing mating system based sporophytic self-incompatibility (SI). Self-incompatibility in
Asteraceae is controlled by a single S locus expressing pollen and pistil allele-specific recognition factors and is subject to negative-frequency-dependent selection, a form of balancing selection that favours new or rare alleles due to their greater population mate availability relative to more common S alleles. Balancing selection typically results in extensive S allele polymorphism over evolutionary time. We investigated the population genetics of self incompatibility in these species to determine how their different population histories have impacted upon their mating system and to examine the interaction between SI and hybridization. Strength of SI was measured and S alleles were identified in population samples of these species and wild hybrids through controlled cross-pollination studies and analysis of subsequent diallel cross results. Data on S allele diversity was combined with molecular genetic marker genotype data to identify the effects of balancing selection on patterns of diversity. These investigations confirmed a reduction in S allele diversity and mate availability in invasive S. squalidus due to the population bottleneck incurred during introduction. However, strength of SI was similar across all study species indicating that sporophytic SI can be robust to extreme population events such as introductions. We found extensive sharing of functional S alleles between species and reduced population structure at the S locus relative to molecular markers indicating that balancing selection at the S locus is an important force for introgressive gene flow between related species.
Evolution of breeding systems in Brassicaceae
Shimizu, KK 1
Kaiser, P 1
, Yasumoto, AA
, Shimizu-Inatsugi, R 1
1 , Tsuchimatsu, T 1 ,
1 University of Zurich, Institute of Plant Biology,
Switzerland
In flowering plants, the transition from outcrossing to predominant selfing through loss of self-compatibility
(SI) is one of the most prevalent evolutionary trends. In many species of Brassica ceae including Arabidopsis , selfing is prevented by the sporophytic SI system, which is controlled by SRK and SCR/SP11, encoding the female and male SI specificity determinants, respectively, at the S-locus. Dozens of highly divergent sequence groups at the S-locus, called S-haplogroups (or
S-haplotypes or S-alleles), confer specificity in selfrecognition. Haplogroups are characterized by low nucleotide diversity within each haplogroup and by high divergence and suppressed recombination between haplogroups. We have studied two genera Arabidopsis and Cardamine , in which the transition to SC occurred many times independently. Cardamine is one of the largest genera in Brassica ceae with >200 species. The prevalence of allopolyploid species suggests the importance of hybridization. To study the reproductive barriers and breeding systems, we conducted reciprocal crossing among 13 species. When outcrossing species are

used as maternal parents, fruit elongation was rarely observed. In contrast, when selfing species serves as maternal parents, seed development tends to start although many of then abort later. The results suggest that the breeding system of maternal parents is critical for the hybrid development, and may be consistent with SI X
SC rule.
Evolution and maintenance of mixed si/sc mating system in
Hypochaeris salzmanniana
DC.
(Asteraceae), an endemic species to the west
Mediterranean
Casimiro-Soriguer, R 1 , Ortiz, MA 1
, Talavera, M 1
, Garcia-Castaño,
, Balao, F 1 , Stuessy, JL 1
TF 3
, Tremetsberger, K 2
, Talavera, S 1
1 University of Seville, Dpto Biología Vegetal y Ecologia
(Area Botanica), Spain; 2 University of Natural Resources and Life Sciences, Vienna, Austria; 3 Institute of Botany,
University of Vienna, Austria
Hypochaeris salzmanniana DC. (Lactuceae, Asteraceae) is an annual herb endemic to the Atlantic coasts of S
Spain and NW Morocco. Previous research has shown that this species exhibits a flexible sporophytic selfincompatibility system, including populations with only self-incompatible individuals, populations with only selfcompatible individuals, and mixed populations with selfincompatible and self-compatible plants. Little is known about the inheritance of self-compatibility and how become to be fixed within the populations. Following different approaches as molecular techniques (AFLPs), demography and reproductive traits (incompatibility tests in the glasshouse and in the wild, pollinator activity) we study the population dynamics, the stability and diversity of the mating systems in natural populations of
Hypochaeris salzmanniana . We found a geographical pattern, from south to north, related with the genetic diversity and the proportion of self-incompatible and self-compatible individuals in the populations. The south side of the distributional area (Morocco) correspond with self-incompatible plants with the highest levels of genetic diversity, probably the ancestral area of the species. The northern populations exhibit different levels of incompatibility in its populations as well as the genetic diversity being lowest in the complete self-compatible populations and vice versa. The establishment of the
Spanish populations probably was originated by migration events from Morocco, when the sea level was lower than today, and the founder effect followed by bottleneck events favoured the appearance and establishment of the self-compatible individuals in the populations. In addition, the pollinators, mainly solitary bees, were efficient enough to get high levels of reproductive success in the wild. Therefore pollinators, at least nowadays, seem to cause no selective pressure on the self-compatible plants.
Are self-incompatible plant species particularly threatened by pollen limitation effects in fragmented populations: what the theory tells us and why empirical approaches are inconclusive?
Vekemans, X 1
1 Université Lille, France
Plant conservation biologists have raised the issue of a potentially greater fragility of self-incompatible species to population fragmentation. This would be caused by a reduction in compatible mate availability due to a lower number of alleles maintained at the self-incompatibility locus (S-locus) in small isolated plant populations.
Indeed, when the number of incompatibility alleles is reduced in a population, a larger fraction of pollen landing on a given pistil will share the same specificity as the pistil, and will thus be rejected. If the pollination service is inefficient for ecological reasons associated to fragmentation, this will lead to an overall reduction in seed production that is expected to be stronger in selfincompatible, as compared to self-compatible species.
This phenomenon is expected to generate a 'mate-finding
Allee effect' specific to self-incompatible species
(sometimes called the' S-Allee effect'), a process generating a positive relationship between reproductive success and population size because of an increase in access to mates in larger populations. Theory predicts that the strength of the S-Allee effect will strongly depends on the genetics of the self-incompatibility system, on the number of extant alleles present in the populations, which itself depends on current and past population sizes and on the rate of gene flow among populations, on the spatial genetic structure at the Slocus, and on ecological attributes of the populations, such as the efficiency of the pollination system.
Empirical studies in the field, however, have failed to demonstrate formally the S-Allee effect. This is because molecular or genetic typing of alleles at the S-locus has rarely been performed in these studies, and also because alternative processes influencing seed production have not been assessed simultaneously. In this presentation, we review ecological genetics theory about selfincompatibility systems in small populations. We present results from experimental approaches aiming at testing the S-Allee effect. Then, we review empirical approaches in the field, and try to highlight the difficulty to disentangle different causes of seed production constriction in small populations. Finally we suggest areas for further investigation, and provide methodological guidelines.
Polyploidy and evolution of breeding systems in
1
Tibetan poppies (
Meconopsis
Vig., Papaveraceae)
Xie, H 1 , Nicotra, A 1 , Ash, J 1 , Cunningham, S 2
Research School of Biology, The Australian National
University, Canberra, Australia; 2 Australian National
Insect Collection, CSIRO Entomology, Canberra,
Australia
The genus Meconopsis Viguier (1814) was erected to contain a single European poppy ( M. cambrica ) that had an elongate style: in contrast to other Papaver species in which the stigma is broad and sessile on the carpels.
Subsequent exploration revealed poppies with a style
(placed in Meconopsis ) in California and especially in the
Himalayan–Tibetan region: including species with large blue, yellow or red corollas that became prized in horticulture. Recent DNA-based phylogenies suggest that the style and flower colour are both variable traits within poppy clades, and Meconopsis now refers only to a clade of about 45 cold-tolerant polyploid Himalayan–Tibetan species. It is arguable that Meconopsis is a subgenus
57

within Papaver . Self-incompatability (SI) is well understood in diploid Papaver (2n=14); however, some polyploid Papaver are self-compatible (SC). The breakdown of incompatability seems necessary for novel polyploids (with no possible mates) to persist, though it may re-evolve. Breeding systems in eight Meconopsis species were tested: species with 2n=56 were SI but those with 2n=76 or 84 were partially SC. Based on phylogenetic reconstructions, some SC species have undergone additional changes: petals enclosing anthers and stigma; a short style; and a shift from blue-violet to yellow or red petals. The SI species that were examined, e.g. M. racemosa , were buzz-pollinated by Bombus or by large Diptera: well suited to outcross pollination. In contrast, enclosed SC species had abundant small Diptera
(e.g. Agromyzids in M. punicea ) that feed and remain within flowers, facilitating self-pollination. We speculate that (1) creation of 2n=76–84 species (from putative
2n=56 + 28 ancestors) was associated with a breakdown in SI, enabling (2) a shift from dependence on large outcross vectors to smaller Dipterans that could live and self-pollinate within flowers, and (3) this led to selection for changes in colour and morphology of flowers that favoured the small Dipterans.
Characterising self-incompatibility in Australian
1 populations of Wild Radish (
Raphanus raphanistrum
)
Mable, B 1 , Newbigin, E 2
University of Glasgow, UK; 2 University of Melbourne,
School of Botany, Australia
Self-incompatibility (SI) in the Brassica ceae is controlled by a single Mendelian locus (the S-locus) but with separate genes coding for male (SCR) and female (SRK) recognition proteins. These genes must be maintained in tight linkage to one another to maintain the ability to recognize self-related proteins in order to prevent selffertilization. Due to this lack of recombination at the Slocus and the 'obligate outcrossing' resulting from having a genetically controlled SI system, high inbreeding depression is expected if the system breaks down to allow self-fertilization. Nevertheless, transitions from outcrossing to inbreeding are frequent between, but also within, species. Although mixed mating used to be considered to be evolutionarily unstable, it is now clear that even in species with genetically controlled SI systems, variation in outcrossing rates occurs across populations in many species. In addition, in some species there is variation in the strength of SI among individuals within populations. In Arabidopsis lyrata , we have found that some populations that maintain high outcrossing rates contain a mixture of self-incompatible (SI) and selfcompatible (SC) individuals, while others contain mostly
SC individuals and are highly inbreeding. This emphasizes that a shift to inbreeding in species with a genetically controlled SI system is a two-step process, with different types of selection pressures potentially operating during the two phases. Deciphering the mechanisms for loss of SI can thus be obscured by subsequent changes associated with selection for inbreeding. In addition, the ‘selfing syndrome’ that is often observed in inbreeding species may evolve later to improve the efficiency of inbreeding. We have been using A. lyrata as a model to understand the genetic and ecological causes and consequences of this multi-phase transition from SI to inbreeding.
Incorporating genetics in the population viability analysis of threatened plants
Gerard Oostermeijer 1
NK 2 , Edwards, GE 3
, Voznesenskaya, EV 2 , Koteyeva,
2
1 Institute for Biodiversity and Ecosystem Dynamics,
University of Amsterda, Amsterdam, Netherlands;
Laboratory of Anatomy and Morphology, VL Komarov
Botanical Institute of RAS, Russia; 3 School of Biological
Sciences, Washington State University, Pullman, USA
Already in the 1980s, pioneer conservation biologists discussed the possibly important role of genetics in the viability of small and isolated populations. Since then, an enormous amount of research has shown that such populations are experiencing loss of genetic variation through drift and inbreeding, and that gene flow is often insufficient to compensate for that. However, when it comes to the question how detrimental this is to their viability, we are still not able to present very many empirical studies that clearly demonstrate this. This is largely due to our failure to effectively integrate genetics with other important components of plant population viability, such as demography and reproductive biology.
Nevertheless, the studies performed on each of these components separately suggest that there are major changes in each of them. The still painfully scarce studies that have attempted to integrate them into single PVAs show that demography, reproduction and genetics act synergistically, and cause the often mentioned but rarely demonstrated extinction vortex. As a result of habitat destruction and fragmentation, many plant populations are genetically depauperate, and show reduced viability that cannot be alleviated by habitat management and restoration alone and requires additional genetic rescue measures. Our knowledge of the implications of outbreeding depression after genetic rescue is still insufficient to provide conservation managers with scientifically sound advice. The rapid developments in ecogenomics will provide exciting new avenues of implementing genetics into plant conservation.
Nevertheless, we still need to combine these new approaches with appropriate field experiments in order to obtain the best answers to still urgent questions. To better implement genetics into practical conservation, researchers need to (i) pay more attention to translating their scientific results, and (ii) focus on the specific questions of conservation managers.
Molecules, monitoring and modelling: integrating genetic and demographic data into conservation planning for the grassland endemic
Rutidosis leptorrhynchoides
Young, A 1
S 4
, Murray, B 2
, Dudash, M 5
, Thrall, P 1 , Pickup, M 3 , Hoebee,
58

1 CSIRO, Canberra, Australia;
New Zealand; 3
2 University of Auckland,
University of Toronto, Canada;
Trobe University, Bundoora, Australia; 5
4 La
University of
Maryland, USA
Effective management of endangered plants must account for both genetic and demographic limitations to population viability and local species persistence that may compromise long term conservation outcomes. The endangered daisy Rutidosis leptorrhynchoides is endemic to the temperate grasslands of south-eastern Australia where it has been subject to severe habitat loss. This species has been the subject of a long term research study combining analysis of reproductive ecology, population demography, population genetics of S allele diversity, cytogenetic variability and outbreeding depression in an effort to understand population trajectories and develop active conservation strategies. These results are integrated in a simulation modelling framework to provide predictions of population viability, identify key threatening processes, and provide management options that maximise the likelihood of long-term maintenance of current populations. Implications for the establishment of new populations are also discussed.
Conservation implications of recognizing polyploid plants: are more species better?
Severns, P 1
1 Washington State University–Vancouver, USA
Small population size, inbreeding depression, and the intentional inflation of within population genetic diversity to cope with impending global climate change induced habitat modifications, are common rationales for within species, inter-population lineage transfer. Assisted migration and intentional between population lineage transfer (from seeds and transplants) carries the possibility of introducing incompatible, as well as, beneficial, compatible genotypes. Conventionally, the primary concern with inter-population lineage mixing has been the unintentional introduction of incompatible genotypes that, upon crossing, produce progeny with lower relative fitness and vigor than crosses within either parental population. Outbreeding depression can yield comparable reductions in plant fitness and vigor to inbreeding depression, but crosses between
‘intraspecific’ individuals that differ in ploidy (and other types of cytological variants) commonly produce sterile progeny, defining them as separate biological species.
With the well-developed genetic barriers that isolate ploidal variants from each other, conservation genetics research into the landscape patterns of chromosome number variation should be common, yet it is not. In this talk, I will present evidence that polyploid species are likely to be more common in protected, rare plants than realized, that the delimitation of polyploid species is important research if rare plant persistence depends on reintroduction and supplemental plantings, and that the formal recognition of polyploid species could enhance rare plant conservation despite policy tradeoffs and an increase in taxonomic complexity.
Are island endemics characterized by low levels of genetic diversity? A case study from the Canary
Islands
Meloni, M 1 , Reid, A
Palacios, JM 4
2 , Caujapé-Castells, J 3
, Soto Medina, M 3 , Conti, E 1
, Fernández-
1 University of Zurich, Institute for Systematic Botany,
Zurich (Switzerland); 2 Institute of Integrative Biology,
ETH Zentrum, Zurich, Switzerland; 3 Jardin Botanico
Canario 'Viera y Clavijo', Unidad Asociada CSIC, Las
Palmas de Gran Canaria, Spain; 4 Island Ecology and
Biogeography Research Group, Universidad de La
Laguna, Tenerife, Islas Canarias, Spain
Islands constitute unique ecosystems, often comprising species found nowhere else on Earth. These endemics, characterized by a unique evolutionary history, represent irreplaceable natural treasures. Island endemics are often rare and/or endangered. One of the reasons for the susceptibility of island endemics to extinction is the low level of genetic diversity that is thought to characterize these species. Genetic variation allows populations to evolve in response to environmental changes; low levels of this ‘evolutionary potential’ limit the ability of insular species to genetically adapt, making them more prone to endangerment and extinction. While there is a trend for insular/rare species to exhibit reduced genetic diversity, recent studies show that island endemics exhibit equivalent levels of genetic diversity compared to closely related, widespread species. Therefore, an extensive knowledge of the amount and distribution of genetic diversity of endemic species and the mechanisms that underlay this diversity are fundamental for their conservation. A population genetic analysis is being performed on three species of the genus Ruta endemic to the Canary Islands. Newly developed SSR
(microsatellites) markers are used to investigate the genetic structure and diversity at both the intra- and interspecific level. By studying the patterns of genetic variation within and between islands, we provide important information on the genetic effects of isolation and small population size on the evolution of insular species. In addition, our results will afford insight on the possible consequences of human-driven processes (i.e. habitat fragmentation, species invasion in response to global warming, etc.) that share some similarities with the evolutionary processes of island colonization.
Finally, knowledge on the genetic variability of endemic species is fundamental for maintaining species’ survival in a changing environment and to direct conservation programs. Águedo Marrero, Ricardo Mesa-Coello and
Félix Manuel Medina contributed samples that were necessary for the molecular genetic analyses performed for this project.
Conservation and genetics of
Acacia
in arid NSW – the tyranny of distance, climate and clonality
Denham, AJ 1 , Ayre, DJ 2
Roberts, DG 2 , Gilpin, A-M
, O'Brien, EK
2
3 , Forrest, CN 2 ,
2
1 Office of Environment and Heritage (NSW), Australia;
University of Wollongong, Australia; 3 University of
Bristol, UK
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Active conservation of arid zone plant species is often necessary to prevent their declines due to chronic recruitment failure. This is largely attributed to direct effects of grazing pressure from domestic, feral and native herbivores and other anthropogenic impacts like soil erosion and climate change. In western NSW some acacias are keystone species – a major structural component of vegetation and thus are likely to be integral to ecosystem function and biodiversity. Management of grazing impacts alone has rarely led to recovery from population declines in these species. Compounding this recruitment failure is the fact that seed production has rarely been observed in many of these acacias, while clonal reproduction of root suckers is common.
Reproductive failure probably reflects either disruption of normal pollination systems or genetic effects of reduced population size and increased isolation.
Nevertheless, it is possible that in stable environments, clonality has been favoured by selection because it replicates successful (i.e., locally adapted) genotypes and episodes of sexual reproduction are naturally rare.
However, sexual reproduction with genetic re-assortment and seed production facilitating dispersal and colonisation should be valuable attributes in the face of significant climate change. Our research programme will attempt to unravel potential genetic and pollination bottlenecks in a number of arid zone Acacia species with a view to facilitating or increasing seed production. We first aim to utilize controlled breeding experiments determine if reproductive success is currently limited either by severe inbreeding depression (in selfcompatible species) or by lack of variation at histocompatibility loci (in outcrossing species), and to compare the diversity of pollinators and patterns of pollinator behaviour for populations and species that vary in their level of seed production. We will also use genetic surveys to determine if populations with little or no seed production are generally lacking in variation at nuclear gene loci and ultimately aim to determine when and how genetic rescue techniques may be applied to promote self-sustaining populations. To date we have begun genetic surveys of several Acacia species including
Acacia carneorum (federally listed as Vulnerable), A. homalophylla/melvillei (an Endangered Ecological
Community in NSW), A. ligulata (not considered at risk),
A. loderi (an EEC in NSW), and A. pendula (a federally listed EEC in part of its range). AFLP surveys of genetic variation in A. carneorum found that all populations surveyed were genetically distinct, implying that outcross pollinations would enhance genetic variation in recipient populations. There was no consistent trend of within population genetic diversity and natural seed set, but the two fruiting populations did have intermediate and high levels of polymorphic loci. We have commenced pollination studies of these species and found that for the relatively successful A. ligulata , natural flower visitation and pollen harvesting is dominated by the exotic honeybee Apis mellifera , resulting in high levels of self pollen transfer. However, hand pollination trials involving outcross pollen transfers both within and among populations have yielded good levels of seed production.
Extended genetic effects of a forest dominant: the case of
Eucalyptus globulus
Potts, B 1
B 1
, O’Reilly-Wapstra, J 1
, Freeman, J
Whitham, T 2
1 , Hamilton, M
, Vaillancourt, R 1
1
, Barbour, R 1
, Bailey, J 1
, Gosney,
, Schweitzer, J 1 ,
1 University of Tasmania, Australia; 2 Dept of Biological
Sciences, and The Merriam-Powell Center for
Environmental Research, Northern Arizona University,
USA
There is increasing evidence that genetic variation in a foundation species may have significant flow-on effects to the associated biota and even to ecosystem processes.
Such extended genetic effects are no better seen in forest trees which dominate many terrestrial ecosystems and provide habitat for numerous dependent organisms. We have been studying these extended genetic effects in
Eucalyptus globulus , a dominant tree of lowland forests of south-eastern Australia. While native to Australia, the species is widely grown in plantations in temperate regions of the world and its molecular and quantitative genetic diversity is amongst the most extensively studied of any forest tree species. An extensive network of progeny trials from large, range-wide open-pollinated seed collections from native races and sub-races of E. globulus have been established for breeding and research purposes. The natural colonization of these pedigreed field trials by local fungi, insect and marsupial species have provided a robust experimental system in which to study individual species, as well as community and ecosystem level responses to genetic variation in E. globulus . This talk will over view the insights we have gained on the extended genetic effects on canopy and bark communities on E. globulus as well as more distal effects on litter and log decay communities. Community responses have been explored at multiple genetic levels from genetically divergent geographic races, additive genetic effects within races, to within family QTL effects. We address the genetic drivers of insect, marsupial and fungal responses to this genetic variation at the species and community level, the relative importance of direct and indirect genetic effects, and the potential for co-evolution through genetic co-variance amongst dependent organisms and feedbacks on tree fitness.
Historical evolutionary dynamics results in convergent ecological consequences for biodiversity and ecosystem function
Bailey, J 1
1 University of Tasmania, Australia
Understanding how historical evolutionary dynamics impact contemporary ecological interactions represents a major frontier linking ecology and evolution. Whether genetic variation in one species can affect associated species interactions and the ecosystem processes mediated by those interactions is fundamental to
60

understanding the links between ecological and evolutionary dynamics. Over the last decade, an emerging body of work provides support for the hypothesis that genetic variation in one species can have extended consequences for associated species. These genetically-based species interactions can also result in significant variation in biodiversity as well as patterns of energy flow and nutrient cycling. Building upon the links between genes and ecosystems, emerging studies suggest that evolutionary divergence in one species can also alter community structure and nutrient cycling. While these studies show that historic evolutionary patterns of race formation can impact biodiversity and soil nutrient availability, it is difficult to determine whether the ecological effects that have been observed are an idiosyncratic consequence of historic evolutionary dynamics or do similar evolutionary processes lead to convergent ecological outcomes? Here we show that parallel evolutionary dynamics can result in convergent ecological outcomes.
Parasitic plants – puppets or puppet masters of community genetic effects?
Rowntree, J 1
1 University of York, UK
Rhinanthus is a genus of hemi-parasitic plants (yellow rattles) common in the grasslands of Europe and North
America. Species of Rhinanthus are generalist parasites with a wide range of documented host plants, particularly associated with grasses and legumes. They have been described as keystone ecosystem engineers, capable of driving change not only in the diversity and structure of associated plant communities, but also the structure of arthropod and soil microbial communities and the cycling of nutrients through the system. Until recently, the role of genetic diversity in these complex grassland systems has been ignored. In greenhouse experiments, we have demonstrated that genetic diversity within a grass host species, and among populations of Rhinanthus , alters the suitability of an individual host plant and the impact of infection by the parasite. We have also shown that parasitic infection of a host grass changes intraspecific competition among aphid herbivores feeding exclusively on the host, while genetic diversity among aphids determines survival of the parasitic plant. Both the effects of the parasitic plant on the aphids and the aphids on the parasitic plant are mediated via genetic diversity in the host plants themselves. Therefore, within species genetic variation in all interacting species (host plants, parasitic plants and herbivores) determines the outcome at various levels of the tripartite interaction. So, are the parasitic plants really driving community level changes or is their success controlled by indirect interactions with herbivores? Current work focuses on teasing out and determining the importance of community genetic effects in natural populations of Rhinanthus .
Genotype by genotype interactions: above- and belowground consequences in
Solidago
Genung, M 1
1 University of Tennessee, USA
Community genetics research has emphasized the idea that species are embedded in a matrix with many other species in variable environments, and this perspective suggests that the effects of genetic variation should not only be assessed in a focal species but also in the neighboring species with which the focal species interacts. Plant–neighbor interactions represent a common type of genotype by environment interaction in which the ‘environment’ (e.g., neighbor plant) contains genes. Additionally, in plant–neighbor interactions participants are fixed in space and forced to interact for resources. Therefore, genetic variation in an individual’s neighbors can have important consequences for overall plant fitness and performance. However, plant–neighbor studies which have examined genotype by genotype interactions have typically focused on the aboveground productivity and fitness of the plants involved, and less is known about how these interactions affect belowground traits and associated communities. We used common garden experiments which manipulated genotype identity and neighbor genotype identity to examine the effects of genotype by genotype interactions on a range of plant traits, including rhizome biomass, coarse root biomass, vegetative biomass, and floral biomass. The effects of plant genotype identity were stronger for aboveground traits than for belowground traits, while the opposite pattern was observed for the effects of neighbor genotype identity. These results show that genotypic variation in a plant’s neighbors can have significant ecological consequences, and that studies which only examine aboveground biomass may underestimate the importance of plant–neighbor interactions.
Genetic feedbacks impact ecosystem processes:
1 linkages between above and belowground processes
Schweitzer, J 1 , Bailey, J 1 , Madritch, M
University of Tasmania, Australia; 2
2
Appalachiain State
University, USA
A growing interest in the consequences of biodiversity on ecosystem processes has led to researchers examining the effects of intraspecific variation on energy flow and nutrient cycles. Fine-scale intra-specific variation in plant traits among genotypes, evolutionarily diverged plants or genetic variation at the population level leads to phenotypic variation, such as differences in productivity and phytochemistry. Across multiple plant groups this variation often leads to heritable organic matter dynamics in the soil system by having large effects on rates of litter decay, influences the composition and activity of heterotrophic soil microorganisms and influence rates of carbon and nitrogen cycling. When seedlings from across a genetic gradient as well as individual genotypes are planted in soils previously conditioned with other species or genotypes, a home-field advantage occurs whereby seedlings have 2-fold higher survival, perform up to 35% better and maintain higher rates of genetic variance in their home soil than in soil conditioned by other plants.
These data suggest that there may be evolutionary consequences for tight plant-soil linkages and that positive feedback may have a role in maintaining plant populations and community dynamics in forest ecosystems.
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Conservation issues associated with the community genetics of foundation forest trees as drivers of community diversity, structure, stability, and evolution
Whitham, T 1
1 Northern Arizona University, USA
The community phenotypes and genetic structure of foundation species, often forest trees, are especially important to quantify as these species are by definition,
‘community and ecosystem drivers’. Using examples from Populus and Pinus , our findings show that different tree genotypes support different communities of organisms (soil microbes, mycorrhizae, arthropods, vertebrates, understory plants, lichens, pathogens) and that these differences can be quantified as heritable plant traits. Thus, genetic diversity in foundation tree species affects biodiversity and is very important to conserve even when these trees are extremely common on the landscape. Using diverse examples, the community phenotypes of forest trees can be traced from the individuals possessing the trait, to the community, and to ecosystem processes such as leaf litter decomposition and N mineralization. Any agent of selection such as climate change that affects the distribution and genetic structure of foundation trees is likely to have cascading impacts, both ecological and evolutionary, on the rest of the ecosystem. Such a community genetics approach allows us to place community and ecosystem ecology within an evolutionary framework, and make better management decisions concerning conservation, biodiversity, climate change, and genetic engineering.
Distribution of plants with extrafloral nectaries
, Weber, MG 2
1
Keeler, KH 1
School of Biological Sciences, University of Nebraska–
Lincoln, USA; 2 Dept of Ecology and Evolutionary
Biology, Cornell University, Ithaca, NY, USA
Extrafloral nectaries (EFNs) are plant glands that secrete sugar water and are located outside the flower. Most studies of their function show them attracting invertebrate predators whose presence reduces herbivory.
EFNs have been reported on a few ferns and in diverse
Angiosperms but no Gymnosperms. The actual number of plant species with EFNs is unknown but more than
4000 species, from over 750 genera in 117 plant families have been reported, about 1.9% of vascular plants.
Plotting the known plant families with EFNs in the angiosperm phylogeny (P. F. Stevens, Angiosperm
Phylogeny Website, Version 9, May 2010) shows EFNs widely but irregularly scattered throughout the Orders.
They are uncommon in the basal Magnoliid families, although present in the Aristolochiaceae. EFNs are infrequent in monocotyledons, yet found on a few grasses, among the Liliaceae (sensu latu), and frequent on the pedicels and peduncles of orchids. Basal Eudicot families such as the Proteales and Gunnerales have no
62 reported EFNs, although they are present within the
Ranunculaceae. Core Eudicots include many families, genera and species with EFNs, including species within the Asterales, Lamiales and Gentianales although EFNs have not been reported in the Apiales. Within some
Orders and a number of families, clustering allows use of
EFNs as taxonomic characters, as within the Fabaceae. In some families, e.g. the Ebenaceae, they are present on all species. Having all or nearly all species with EFNs is most common within genera, e.g., Passiflora
(Passifloraceae), not at higher levels of classification.
The data is consistent with separate origins of EFNs in different lineages (e.g., Viburnum , Adoxaceae) and likewise with loss of the trait in lineages within some taxa where EFNs are otherwise common (e.g. Ipomoea ,
Convolvulaceae). EFNs are morphologically diverse and can occur on virtually every aboveground plant tissue.
They have not been reported underground. All life forms have EFNs, but they seem particularly common in vines and particularly uncommon in annual herbs. EFNs are largely if not entirely absent from aquatic plants. EFNs appear to be more common in tropical taxa than temperate ones, even allowing for the greater species richness in the tropics. Considering that EFNs release water, they are surprisingly common on desert plants.
The duration of activity and the complexity of EFN structure varies greatly between taxa. The diversity of the morphology and distribution of EFNs suggests a strong role of selection on them, but the joint impact of phylogeny and ecology has made recognition of patterns difficult. New technologies and better phylogenies offer much hope for new insights in the coming decades.
Nectaries in ferns: their distribution and ecological
1 role in pteridophytes
Koptur, S 1
Florida International University, USA
The oldest group of plants in which nectar secretions have been observed are the pteridophytes; nectaries have been reported in a dozen extant genera. The function of these nectaries has been investigated in several fern species, and in some circumstances has been demonstrated to have an antiherbivore role, attracting and maintaining biotic defense (ants and/or other predatory arthropods). The widespread bracken fern, Pteridium aquilinum , has been studied in many places around the world; the benefit from the nectaries varies with geographic location, nectary visitors involved, and array of herbivores present in each location. More experimental work is needed in genera with nectaries about which we have little ecological information, especially about their plant/animal interactions. Nectaries in ferns are most often morphologically simple, and may be overlooked if the exudates are removed by various visitors in nature. When plants are grown in a greenhouse, or in protected areas such as gardens or nurseries where potential nectar drinkers are absent for various reasons, one may observe either droplets of nectar, or sooty mold that grows on, and consumes, accumulated nectar. The presence of sooty mold may be the first indication that nectar is being produced (once honeydew from phloem-feeding insects is ruled out), and then a simple sugar detection method can reveal if nectar is the reason for the sooty mold accumulation. Sooty

mold occurs at the base of fronds of Pleopeltis crassinervata , a widespread epiphyte growing on a variety of trees found in cloud forest areas of Veracruz,
Mexico. This species, segregated from the former genus
Polypodium , is not in the section of Polypodium known to possess leaf nectaries, and so is a new record for this species, and this group of ferns. Previous experimental work on other epiphytic species of Polypodium demonstrated the protective role of ants for developing fronds, so we conducted similar experiments (using nylon polish to cover nectaries, rather than excluding ants as in earlier work) as the fronds of Pleopeltis crassinervata are small, and often lie flat against the trees upon which they grow. Fronds with nectaries covered suffered greater damage from herbivores over the course of their development, compared with fronds that had nectaries functioning normally. A parallel experiment conducted on a sympatric fern species without nectaries showed no difference between manipulated and control fronds. Seven species of ants were observed visiting nectaries and on fronds of P. crassinervata , but differed in their response to herbivore larvae placed on the ferns.
The presence of nectaries, and the relationship with ants, may explain why this epiphytic fern is the most widespread and persistent species, resilient to disturbance, and found on shade trees in coffee plantations throughout its range.
Diversity and evolution of extrafloral nectaries in
Leguminosae
1
Marazzi, B
Salinas, A 3
1 , Bronstein, JL
, Luckow, MA
2
4
1 , Simon, M
, Sanderson, MS 1
2 , Delgado
Dept of Ecology And Evolutionary Biology, University of Arizona, USA; Embrapa Genetic Resources and
Biotechnology, Brasília, Brazil; 3 Instituto de Biología,
4
Universidad Nacional Autónoma de México, Mexico;
Dept of Plant Biology, Cornell University, Ithaca, New
York, USA
Extrafloral nectaries (EFNs) are nectar-secreting organs on plant parts external to flowers, which commonly attract ants that feed on the nectar and, in return, protect the plant from herbivores. EFNs occur in up to 117 angiosperm families, and are common in Leguminosae.
Not only is Leguminosae the third largest and economically the second most important angiosperm family, but it also dominates many kinds of vegetation worldwide and is highly diverse in its interactions with ants and other organisms. Indeed, ant-legume interactions may play an important role in shaping the ecology and evolution of entire ecosystems. Leguminosae display an impressive diversity in EFN morphology, location on the plant, nectar features, and ecological role. The phylogenetic distribution and evolution of EFNs have not yet been characterized for this family. Here we explore:
How many legume genera have at least some species with EFNs? How many times and when did EFNs evolve in legumes? Of the 105–116 legume genera with EFNs
(in at least some species), we compiled, primarily from published reports, 46–47 belonging to the subfamily
Mimosoideae, 32–37 to the Papilionioideae, and 27–32 to the Caesalpinioideae grade. Within subfamilies, EFN genera represent only 7–8% of all papilionoid genera, whereas caesalpinioid and mimosoid EFN genera represent up to 16–19% and 56–57% of their genera, respectively. The phylogenetic distribution of EFNs in
Leguminosae thus appears to be diffuse, dense, and patchy simultaneously, with most EFN genera – and probably also most EFN species – concentrated in the sister clade of the Papilionoideae, which includes many caesalpinioid lineages and the Mimosoideae. Foliar and stipular EFNs appear to be most common (ca. 65% and
15% of the genera). Parsimony reconstruction of EFN presence reports mapped on a Bayesian mega-phylogeny of 829 legumes (ca. 400 genera) inferred at least 35 independent EFN origins (not considering reversals or gains after losses). While EFNs seem to have originated multiple times in the Caesalpinioideae grade and
Papilionoideae (at least 24 and 10 times, respectively), they appear to have originated only once in
Mimosoideae. Based on Bayesian divergence time analyses of the legume mega-phylogeny, we present results of the tempo and mode of EFN evolution in legumes. We also explore implications of our results for the current definition(s) of EFNs in Leguminosae and the associated levels of homology/homoplasy. Our findings provide new insights that increase our understanding of the evolution of EFNs in Leguminosae, as well as of the natural history of ant-plant associations and ant-mediated anti-herbivore defense strategies.
Passiflora
as a model system for studying nectary diversification: insights and implications
Krosnick, S 1 , Gasser, C 2 , Potter, D 3
1 Dept of Biology, Southern Arkansas University,
Magnolia, USA; 2 Dept of Molecular and Cellular
Biology, University of California, Davis, USA; 3 Dept of
Plant Sciences, University of California, Davis, USA
Passiflora is a diverse genus consisting of more than 520 species of vines, lianas and small trees. The group displays remarkable variation in floral and vegetative features, even among closely related species. A key characteristic of the genus is the presence of complex nectaries throughout the plant. These nectaries encourage pollination, deter herbivores, and attract bodyguards.
Floral and extrafloral nectaries are present on all species of Passiflora within flowers, on floral bracts, stipules, petioles, on leaf margins, and abaxial leaf surfaces. In
Passiflora , the position, shape, size, and arrangement of nectaries is so variable that homology assessment among the different nectary classes is difficult. As nectaries likely represent a key morphological innovation for the genus, the need for clear statements of homology among the different nectary classes is great. Examination of nectary form and function in Passiflora has yielded several important insights regarding the evolution of nectaries in the genus. First, initial hypotheses of homology between floral and extrafloral nectaries have been rejected based on molecular and morphological data. This is in contrast to the situation in other Eudicots, where the two types of nectary have been shown to share molecular similarities. Second, the use of nectar sugar profile data to assess variation among extrafloral nectary classes has not yet yielded informative differences in sucrose, glucose, or fructose. Amino acid content may be of greater importance for extrafloral nectaries compared to floral nectaries. Nectar secretion occurs much earlier in development than originally expected and appears to be passive in the mode of release. This calls into question
63

earlier accounts citing cuticle rupture as the primary mode of nectar release. Third, scanning electron microscopy has revealed a unique thinning pattern in the cuticle of petiolar nectaries. These thinning areas eventually develop into pore-like structures in the cuticle, a relatively uncommon mode of nectar release in angiosperms. Fourth, anatomical and developmental data collected thus far suggest similarities between glandular leaf teeth found on the margins of stipules and the lamina with those found on the petiole and abaxial surface.
These data suggest that a similar developmental program is acting during leaf development to create diverse kinds of extrafloral nectaries. Thus, the final location and form of the nectaries may depend most on the maturity of the leaf tissue that this shared developmental program is active within. These data are considered with regard to the overall phylogenetic framework of Passiflora . The patterns of diversification in extrafloral nectaries are closely tied to the lineages in which they are found, supporting a preliminary hypothesis that nectaries do in fact represent a key innovation for the genus.
Implications for homology assessment are considered not only for Passiflora , but for other relevant angiosperm lineages as well.
Glands and geography: geographic variation in extrafloral nectaries in the genus
Adriana
(Euphorbiaceae)
Mackay, D 1 , Whalen, M 1
1 Flinders University, Adelaide, Australia
Plants in at least 93 families possess extra-floral nectaries
(EFNs) that secrete nectar that is attractive to ants
(Koptur 1992). Plants in a wide range of habitats that attract ants to their foliage by providing nectar from
EFNs often experience reduced herbivore loads or levels of herbivory. Temporal and spatial variation in the quality of the defence provided by ants against herbivores has also been noted by various authors and has been ascribed to variation in weather conditions, altitude, the density and diversity of ants and herbivores and the quality of nectar rewards (Koptur 1992, Bluthgen et al. 2004, Rico-Gray and Oliveira 2007). To further investigate factors influencing variability in the outcomes of this association, we examined geographic variation in the associations between ants and plants of Adriana , a genus of dioecious shrubs which is widely distributed in
Australia. Both male and female plants possess stipular
EFNs at the bases of the petioles and nectaries on the floral bracts. Here we review and discuss a series of studies examining the associations between ants and
Adriana plants across a continental scale. Consequences of ant visitation to male and female plants were studied at sites near Broome, Western Australia; Toowoomba in south-east Queensland; east Gippsland in Victoria and near Adelaide in South Australia. Ant and herbivore communities on adrianas varied among geographic locations, as did the effects of experimental ant exclusions. Ants in five to ten genera were observed visiting EFNs at different locations and significant effects of ants in lowering herbivore densities were observed at several locations. Significant effects on levels of foliar herbivory were only detected near
Adelaide and only on male plants which were visited by four times as many ants as were female plants. The
64 studies above were conducted under variable climatic and local conditions, some in periods of drought, and we explore the association between geographic variation in climate and the potential role of EFNs as a plant defence.
The provision of water-based nectar from EFNs may incur greater costs in attracting ant bodyguards under dry conditions, and in general, conditionality in the outcomes of defensive mutualisms may reflect variation in the relative costs of different defences. We examined the effects of variation in climatic variables, including rainfall, on geographic variation in nectary sizes across the ranges of three taxa within Adriana in Australia. For each taxon, there was a significant association between gland size and climatic variables related to moisture availability. A glasshouse experiment testing the effects of water stress on nectary longevity also revealed that secretory activity declined more rapidly on droughtstressed plants. Conditionality in the outcomes of associations between ants and plants with EFNs are discussed in the context of predicted patterns of future rainfall and also in relation to the expected investment by male and female plants in anti-herbivore defences.
Extrafloral nectaries the basis for ant–plant mutualistic networks
Rico-Gray, V 1 , Diaz-Castelazo, C 1
1 Instituto de Ecologia, AC, Xalapa, Mexico
Interspecific interactions evolve as a geographic mosaic, generating not only differences across space and time in traits and ecological outcomes but also in the number of species involved. The network structure of these interactions influences the nature of these ecological and evolutionary outcomes. Mutualistic networks exhibit complex but predictable patterns of interaction, frequently involving many species that form complex networks of interdependence. Recent analyses have shown that mutualistic networks among free-living species are nested. The overall pattern is one of weak and asymmetric links among species. These network patterns may be explained by ecological variables and evolutionary history but few data exist on their spatial and temporal stability. Several questions have been addressed using ant–plant interactions: Are ant–plant mutualistic interactions nested? Which ecological conditions may result in differences in nestedness among communities? Does interaction intimacy affect network structure and species richness? Do networks change in time? Most of the results are based on the analysis of the structure of a multispecific network of interacting ants and extrafloral nectary-bearing plants sampled between
May 1989 and April 1991, and again between October
1998 and September 2000 in La Mancha, Veracruz,
Mexico. Data analysis and statistics were performed using: Microsoft Excel, Pajek, Aninhado, UCINET, and
XL-Stat. The structure of ant–plant networks exhibits nested patterns. Interaction intimacy markedly affects the structure of these networks. When interaction intimacy is high, the interactions are compartmentalized, when interaction intimacy is low, they exhibit nested patterns.
These results support a promising approach for the development of multispecies coevolutionary theory, leading to the idea that specialization may coevolve in different but simple ways in antagonistic and mutualistic assemblages, and that simple features of mutualistic

interactions are likely to have important consequences for the coevolutionary process and the patterns it generates in the organization of biodiversity. When compared in time, the nested topology of the network remains similar, group dissimilarity increases, standardized number of interactions for ant species increases and shifts towards higher values for plant species, more ant species and less plant species constituted the core of the more recent network, and presence of invasive/ruderal species increased while their contribution to nestedness remained the same. Generalist species characterized these patterns and appeared to maintain the stability of the network, since the new invasive/ruderal species incorporated in the communities were linked to this core of generalists: the overall network structure remains unmodified.
Where is the greatest diversity of liverworts and where are the threats greatest?
Söderström, L 1
A 3
, Hagborg, A 2 , Von Konrat, M 2 , Séneca,
1 Dept of Biology, Norwegian University of Science and
Technology, Norway; 2
Museum, Chicago, USA;
Dept of Botany, The Field
3 Dept of Biology, Faculty of
Science, University of Porto, Portugal
Bryophytes are important components of the vegetation in many regions of the world, constituting a major part of the biodiversity in moist forest, wetland, mountain and tundra ecosystems. Because of their sensitivity to the immediate microhabitat, these groups of plants are important environmental indicators and have been used as predictors of past climate change, to validate climate models and as potential indicators of global warming.
Yet, alarmingly, our understanding of the degree of threat of these organisms is scant. In order to understand the threat level of bryophytes one needs to know i) where they occur (geographically and habitat) and ii) what the threats are for that region or that habitat. This talk will give a global overview of the liverwort diversity using the Early Land Plants Today (ELPT) database. The diversity will be compared with the threat to habitats using the 'Global 200' Ecoregions as larger scale units of threatened regions. In addition, human population density is added as an additional layer of threat. This will be compared with figures for threats for areas where
IUCN Red Lists occurs to evaluate the overall trends. We also test correlations between high levels of liverwort diversity and existing international priority areas for conservation.
Recent research on bryophyte ecology in relation to climate change
Slack, N 1
1 Biology Dept, The Sage Colleges, Troy, USA
Bryophytes are currently being used in many types of climate change research. Not only global warming but the effects of increased carbon dioxide and ultraviolet light as well as changes in precipitation are being studied, using bryophytes, in many different countries on several continents. Research in a variety of ecosystems from desert to peatlands to rain forest will be discussed. Some of this research has recently been published in Bryophyte
Ecology and Climate Change. In addition our current research on the effect of temperature increase on rare alpine plants including bryophytes in Northeastern U.S. mountains will be reported. The habitat for alpine bryophytes is diminishing and plants from lower elevations are invading the tundra habitat, as is also true in the European Alps.
Nothofagus
as a keystone host for bryophyte diversity
1 in cool temperate rainforest
Dalton, PJ 1
School of Plant Science, University of Tasmania,
Hobart, Australia
Cool temperate rainforest (CTR) in Tasmania is a climax vegetation characterised by the dicot tree species from the genera Nothofagus, Atherosperma and Eucryphia and the conifer tree genera Athrotaxis, Phyllocladus and
Lagarostrobos . In comparison to tropical rainforests,
CTR have fewer woody tree species and the major floristic component is the bryophytes, whose occurrence dominates the epiphytic flora. A total of 61 bryophytes were recorded from the trunk of N. cunninghamii across its distribution in western Tasmania with hepatics outweighing mosses up to 5:1. Most species were facultative epiphytes and the study analysed overall cover, species richness and liverwort to moss ratios.
Results clearly indicate that there was considerable variation and it would appear that microclimatic factors rather than bark characteristics are having the greater influence for a change in composition of the epiphytic flora from one forest system to another. Within a single type of cool temperate rainforest, there is a marked zonation of the epiphytic flora on the major host tree and it is further proposed that moisture/humidity is primarily responsible for the partitioning of bryophyte species. The results of these studies support the hypothesis that
Nothofagus cunninghamii is a keystone species for diversity of bryophytes in cool temperate rainforest.
Conservation of the New Zealand bryoflora
Fife, AJ 1 , Glenny, D 1
1 Allan Herbarium, Lincoln, New Zealand
New Zealand is considered a global hotspot for bryophyte diversity. The latest figures for the number of documented and described species are 534 mosses, 616 hepatics and 13 hornworts. There is a high level of endemism: c. 21% of mosses, c. 53% of hepatics, and c.
60% of hornworts are considered endemic. A panel of
New Zealand specialists has met 3 times since 1992 (and once prior) to evaluate rare and poorly documented bryophyte taxa using a nationally developed 'New
Zealand Threat Classification System'. In the most recent ranking threatened or uncommon status has been applied to 269 described taxa (c. 24% of the total) and to another
25 entities, mostly taxa awaiting description. Forty-four
65

species of bryophytes are considered Threatened, with 28
(12 mosses; 16 hepatics) Nationally Critical, 9 Nationally
Endangered (2 mosses; 7 hepatics); and 7 Nationally
Vulnerable (4 mosses; 3 hepatics). A further 138 taxa and undescribed entities are considered to be At Risk and 135
Data Deficient. Two mosses ( Lindbergia maritima
Lewinsky and Epipterygium opararense Shaw & Fife) and two hepatics ( Petalophyllum australe Colenso and
Schistochila nitidissima R.M.Schust.) are highlighted to illustrate difficulties and successes associated with their listing and with preliminary efforts at active conservation. Each species present unique challenges.
We need to distinguish naturally rare species from those which have become rare through human activities, but sometimes cannot do this confidently. Lindbergia maritima and Schistochila nitidissima illustrate the former condition, while the rarity of Epipterygium opararense and Petalophyllum australe probably results from an interplay of both factors. For species truly in decline, we need to assess the threat/s and plan appropriate management efforts. Such efforts are in their infancy in New Zealand. There is a lack of resources for those who know the plants sufficiently well to conduct the needed surveys, although a limited amount of funding for investigating 'Data Deficient' taxa is now available through the NZ Dept of Conservation. This should assist assessing the main threat types and permit efforts to reverse them for a limited number of taxa. Our most urgent needs are additional surveys and learning how to raise bryophytes in vitro and to transplant them back into the wild.
Conservation of bryophytes in tropical oceanic islands: study case of La Réunion
Ah-Peng, C 1,2 , Bardat, J 3 , Hedderson, T 1 , Strasberg, D 2
1
2
University of Cape Town, Botany Dept, South Africa;
Université de La Réunion, La Réunion; 3 Muséum
National d'Histoire Naturelle, Département de
Systématique et Evolution, Paris, France
Islands are unique systems due to their isolation, restricted area and natural oceanic barriers. Their environments are particularly vulnerable to the impacts of climate change, natural hazards (tropical storms, volcanic eruptions) and the pressures of human activities.
Despite the fundamental importance of their biodiversity for economic development, less interest has been focussed on small organisms. Réunion Island
(Mascarenes archipelago), along with Madagascar and the neighbouring islands forms a hotspot of diversity in the Western Indian Ocean. The island possesses the highest summit in the geographical area, the Piton des
Neiges (3070 m), providing a myriad of habitats along this extensive climatic gradient. Compared to the sister islands, the bryoflora of Réunion has always been more well known especially now with the increase of ecological studies on the island. Presently more than 800 species of bryophytes are reported from Réunion, its diversity is by far dominated by mosses followed by liverworts and hornworts. Interestingly the rate of endemism for bryophytes is low (<10 %) in comparison with the 30 % for the vascular plant flora. Bryophyte richness on the island peaks in the cloud forests (~ 1150 m), which provide favourable habitats for their development. Due to a growing population, especially in
66 the littoral area, some habitats and consequently some bryophytes species are endangered. As a consequence it is necessary and urgent to address which measures should be taken to protect this rich flora. Multiple actions have been initiated with the aim of increasing knowledge and awareness of the diversity and distribution of bryophytes on Réunion. A taxonomic and nomenclatural index was created, a GIS for bryophytes was implemented allowing the distribution of species to be mapped and areas of interest identified both in terms of diversity and where distribution data of bryophytes was lacking. Conservation tools in the form of floras and Red
Lists are a priority for the island. Training in bryophyte identification remains key for enhancing knowledge and interest of the local bryoflora. Indeed, it is important to determine research directions and measures to be taken for these lesser-known plants that, nevertheless, form an important part of the diversity on the island. How do we best conserve this important and diverse bryoflora on such a small island which is threatened by high anthropogenic pressures? In the coming future we wish to use Réunion as a model for conservation actions for bryophytes in similar systems (tropical oceanic islands), setting up a methodology for taking into account bryophytes by nature managers, and promoting target actions to encourage authorities to include bryophytes in their conservation plans. Réunion is part of the European outermost regions and territories, these places have an exceptional biodiversity and the pressures they face make them sentinels of global change, there is a target goal/task to unite actions to slow down biodiversity depletion before even describing it between these different regions (Indian, Atlantic and Pacific Oceans).
What are we trying to conserve? The role of phylogenetics in taxon recognition and prioritization
1 for conservation
Mishler, B 1
University of California, Berkeley, USA
Much greater attention needs to be paid to conservation of bryophytes, and to do this effectively a number of aspects of their biology must be advanced. I will focus on systematic aspects of the problem – what is to be conserved? Many bryophyte taxa, especially in the tropics, are poorly characterized, and the names given them under the current code of nomenclature cannot clearly indicate their meaning and status. Because of many problems with instability and lack of comparability of ranks in the current Linnaean system, we need a new rank-free phylogenetic basis for taxonomy (e.g., the
Phylocode). In such a system, all taxa to be named should be hypothesized monophyletic groups. Not all such groups need be named, but those that are named formally should be given unranked (but hierarchically nested) uninomials. The applicability of a name should be fixed using multiple specifiers rather than a single type specimen as in the current system. What about the species rank? This is controversial even within the
Phylocode community, but I argue that the least inclusive taxon (formally known as 'species') should be treated in the same rank-free manner as taxa at other levels. Current entities ranked as species are not comparable in age, internal genetic diversity, ecology, the amount of morphospace they occupy, the size of the phenetic 'moat'

around them, or the amount of interbreeding within them
(or lack thereof between them), nor can they be made to be comparable through any massive realignment of current usage. Finally, I will explore the practical implications of eliminating the rank of species for such areas as education, ecology, evolution, and conservation, and conclude that these purposes are better served by this move. The basic element of biodiversity is not species, but rather the semaphoront, a specimen along with all associated metadata. These semaphoronts can be built up into hypotheses of clades using molecular and morphological data, to study all aspects of biodiversity science. How can we inventory biodiversity without species? New quantitative measures for phylogenetic biodiversity, which take into account the number of branch points (and possibly branch lengths) that separate two lineages, should be applied. What does ‘rarity’ mean without ranks? The phylogenetic worldview described above can clarify greatly what 'rarity' means. Rarity fundamentally means having few living close relatives, where ‘few’ and ‘close’ are measured quantitatively on cladograms. Conservation priorities can be better guided by phylogenies rather than a list of species names. The need for biodiversity inventories has never been greater; fortunately recent advances have been made in the discovery, characterization, and presentation of biodiversity information. High-throughput molecular techniques, faster phylogenetic methods, highly improved geographic information systems and databases, and better web visualization approaches are all welcome aids in this crucial task. Biodiversity assessment no longer needs to rely on pre-evolutionary, species-centric, list-making approaches, but instead can focus on biodiversity discovery and understanding at many phylogenetic scales. Examples will be presented from the
Moorea Biocode Project in French Polynesia, and the
Jepson Flora Project in California.
Diversity of medicinal plants in Maruthamalai,
Coimbatore District, Tamil Nadu, India
1
Narayanasamy, V 1
Sir Thiagarajar College, Chennai, India
Biodiversity is the totality of genes, species and ecosystems of a region. It is not evenly distributed due to variation in environmental conditions. Diversity consistently measures higher in the tropics and lower in other climatic zones generally. The aesthetic value of biodiversity is well realized and it could not be valued ideally with the current economic valuation models. Of late, biodiversity is more reckoned for its unbelieved potential for supplying novel genes for a variety of applications in biotechnology and pharmacology.
Western Ghats are themselves renowned as one of the world’s great biodiversity hot spots. Maruthamalai, a part of Western Ghats in Tamil Nadu have arid climatic condition with tropical dry deciduous forest vegetation.
In the present study 62 medicinal plants of 32 families were identified. Medicinal plants like Capparis zeylanica, Flueggea leucocephala, Erythroxylon monogynum, Spergula arvensis, Croton bonplandianum,
Cleome viscosa, Mollugo ceruviana, Oldenlandia umbellate, Grewia hirsuta, Pavetta indica, Gardenia resinifera, Eucalyptus longiflorus, Zizyphus trinervia, etc., are used for various ailments such as cough, cold, headache, fever, jaundice, diabetes. Still to bring out the information on therapeutic uses, further studies are suggested in this region.
Ethno-pharmacological studies of medicinal flora in
1
Palamalai, the Western Ghats, Tamil Nadu, India
Srinivasan, N 1
Gurunanac College, Chennai, India
The conventional knowledge of folk-medicine is now deteriorating drastically along with loss of species in many biodiversity nations. The floral diversity of
Palamalai is recognized for conventional therapeutic values. However, no floristic list was prepared with respect to medicinal uses in this region. Therefore, it is necessary to collect and document medicinal plants in
Palamalai. In this regard, a survey on the medicinal plants of Palamalai was made. The study revealed that the plant species such as Commiphora wightii, Grewia obutifolia, Sapindus emarginatus, Cassia auriculata,
Phyllanthus reticulate; Waltheria indica, Plumbago zeylanica, Trichodesma indica, etc., were tested for different pharmacological activities viz., antipyretic, antiulcer, anti-inflammatory, antidiabetic, anticancerous, etc. The study confirmed the traditional on these species for medicinal uses.
Genetic diversity and its conservation implications of two endangered medicinal plants of the Western
Ghats of South India using ISSR markers
John De Britto, A 1
1 PG & Research Dept of Plant Biology and
Biotechnology, St Xavier’s College (Autonomous),
Palayamkottai, Tamilnadu, India
The demands of the majority of the populace for medicinal plants have been met by indiscriminate harvesting of spontaneous flora, including those in forests. This has resulted in severe loss of habitat and genetic diversity. As long as the destruction of forest continues, medicinal and aromatic plants and their natural habitats will remain under the threat of overexploitation than ever before. The ultimate goal of conservation is to preserve the natural habitats of vulnerable medicinal plant species and to achieve their sustainable exploitation in less vulnerable areas.
Conservation genetics plays a major role in guiding relocation and reintroduction efforts and in prioritizing species for conservation. Genetic approaches allow us to assess the variability in these populations, as well as to assess whether there is any history or future danger of loss of genetic variability. Genetics can help to do viability analyses by testing hypotheses concerning how long genetic variation might persist into the future. Two endangered medicinal plants Piper barberi Gamble and
Adhatoda beddomei C.B.Clarke were collected from ten locations in the Western Ghats of south India and the genetic variability and genetic differentiation within each species were examined. Inter-simple sequence repeat
67

(ISSR) marker data were obtained and analyzed with respect to genetic diversity, structure and gene flow.
Natural factors such as unprecedented climatic changes and anthropogenic pressures such as habitat degradation and over harvesting are largely responsible for genetic depletion and rarity of these medicinal plant species.
New means and mechanisms are to be worked out for long-term conservation and sustainable utilization of these rare and economically important medicinal plants.
The populations of both the species urgently need protection to maximize the genetic diversity and diminish further substantial loss within the populations.
Conservation strategies for these two endangered species are proposed. The strong genetic differentiation among populations of these two species indicates that the management for the conservation of genetic variability should aim to preserve every population of these species.
It is suggested that in situ conservation be an important and practical measure for maintaining the genetic diversity of these species. Ex situ conservation should sample from different populations across the distribution range of these species to conserve high genetic diversity.
High altitudinal aromatic plants and their uses as religious purposes by local peoples and bhotias in
Ganga Valley of Uttarkashi District (Uttrakhand),
India
Kumar, S 1
Rather, MA 2
, Dhingra, GK 1 , Arya, R 1 , Rawat, AS 1 ,
1
2
RCU Govt PG College Uttarkashi, Uttarakhan, India;
Dept of Chemistry, RCU Govt PG College Uttarkashi,
Uttarakhand, India
It has been known for a long time that aromatic substances (essential oils, extracts) contained in plants often exert psychological effects ranging from sedative to excitatory actions. Medicinal effects have also been confirmed through numerous experiences. Aromatic compounds from plants for the purpose of affecting a person's mood or health. In ancient times, aromatic trees and herbs were offered to deities, mostly as incenses that were believed to carry people's wishes, such as requests to cure sick people, and to gain Para psychological experience, to Heaven. It therefore may not be surprising that aromatics were often used in medicine, which once had close ties with shamanism, animism and other religious activities. The ingredient or raw materials used in hawan are basically herbs and roots of various aromatic plants. The study area is concentrated around the Ganga (Bhagirathi) Valley (Harsil, Bhagirathi Peak,
Gangotri-1, Gangotri-2, Tapoban, Nandan Ban,
Banderpunch, Satopant and Gowmukh, etc.) of
Uttarkashi district in Uttarakhand state. The purpose of this research work is to describe the importance of aromatic plants, habits, identification and their uses by local peoples/ Bhotias in the form of Dhoop sticks,
Agarbatii sticks and Havan as religious purposes. Plants belonging to
Caryophyllaceae, the families
Chenopodiaceae,
Boraginaceae,
Compositae,
Cruciferae, Gramineae, Labiatae, Leguminosae,
Geraniaceae, Liliaceae, and Rosaceae were reported highly aromatic in nature at research sites. In study areas
Geranium himalayense, Ocimum bacillicum, Lavandula angustifolia, Cymbopogon Citratus Eucalyptus globulus,
Rosmarinus officinalis, Actinopteris dichotoma, Skimmia
68 laureola, Skimmia anquetelia and Artemisia sp., etc, were recorded. Local peoples and Bhotia tribes used these plants to made agarbattis, Dhoop and Havan materials, etc. Powdered gummy wood as an adhesive medium for binding the paste to the stick. The use of hands and fingers has resulted in the hand rolling process as no machines were used since ancient times.
Traversing the continent: local adaptation in poplar
(
Populus trichocarpa
) along western North America
1 geographic clines
McKown, AD 1 , Guy, RD 1
University of British Columbia, Vancouver, Canada
Cottonwood poplar ( Populus trichocarpa ) is a widespread, deciduous tree in western North America that shows local adaptation and selection for traits in relation to local growing season and climate. The genomically sequenced tree is a model for understanding tree biology, and there is strong interest for its use in the agro-biofuels industry. Over 2000 trees (representing replicate plantings of 499 collected genotypes in 150 provenance localities from Alaska to Oregon) were grown in a common garden in Vancouver, Canada. Our studies investigated inherent trait variation within P. trichocarpa focusing on numerous traits relating to seasonal (phenological) events, biomass accumulation and growth rates, C and N isotopes, and photosynthetic traits. Traits were analyzed in relation to photoperiodic
(latitudinal) and climatic (temperature, precipitation) clines, and in trait-trait combinations to understand relationships between plant structure and plant performance. Collaborative work using single nucleotide polymorphism data showed that there is no population structure in P. trichocarpa across western North
America; however, there is evidence for selection in some genes suggesting local adaptation. Our findings showed that trees grown in a common garden still demonstrate substantial differences in phenological events, growth rate, biomass accumulation, C and N isotopes, and photosynthesis for the different genotypes.
Many traits had strong correlations with latitude, and some showed weak association with climate parameters.
In general, the northern genotypes tended to show faster incipient growth rates, higher photosynthesis and greater stomatal conductance. Total biomass in these trees was far lower than southern genotypes because northern trees set bud entering ‘fall phase’ earlier and lost their leaves sooner than southern genotypes. Thus, despite faster rates of growth and higher photosynthesis in northern genotypes, the innate response to photoperiod shortened the active growing season resulting in smaller trees with less accumulation in biomass. Southern trees showed the opposite trends, and generally showed higher water use efficiency, lower leaf mass per area, and lower nitrogen use efficiency. Numerous traits measured in this study point to response to photoperiod (i.e. latitude) as a strong driver of local adaptation in P. trichocarpa , and multiple lines of evidence from our common garden experiment suggest that northern genotypes show selection for

specific traits relating to maximizing a shortened active growing season, compared to southern genotypes. Thus, northern trees of P. trichocarpa are intrinsically adapted for a short growing season by adopting fast growth rates and investing in short-lived, high-output leaves, whereas southern genotypes of the same species are adapted for a longer growing season overall.
Diversity patterns of plant dispersal syndromes found in Colombian tropical forests, and their potential biotic and environmental determinants
Correa-Gómez, DF 1 , Stevenson, P 1 , Álvarez-Dávila, E 2
1 Centro de Investigaciones Ecológicas de la Macarena
(CIEM), Bogota, Colombia; 2 Instituto de Investigaciones
Alexander von Humboldt (IAvH), Colombia
Dispersal syndromes are key functional traits found in every species of vascular plants. Because of their importance for plant recruitment, they help us to uncover the complex relationships found in tropical forests, and in a similar way to structural diversity and floristic composition, they show patterns along environmental and anthropogenic gradients. Through this study, we evaluated the possible relationships found between dispersal syndromes and biotic and environmental factors, using the largest database of vascular plants found in permanent vegetation plots in Colombia (101 permanent plots, equivalent to 101 hectares). The plots are found throughout the different biogeographic regions of Colombia (Pacífico, Amazonas, Orinoco, Magdalena
Medio, Andes, Magdalena Alto, and Caribe), and include vascular plants with diameter at breast height (DBH) equal or higher than 10 cm. Based on morphological features, dispersal syndromes were assigned to each species, calculating their abundance and percentage per plot. Preliminary results show that forest fragmentation, flooding influence, wind speed and annual mean rainfall are related with the observed dispersal syndromes trends.
Sinzoochory was negatively related with forest fragmentation, hydrochory increased in flooded areas, anemochory increased in windy areas and endozoochory diminished from wet to dry regions. It is proposed that further spatial correlation analysis, as well as plants phylogenetical relationships are needed in order to correctly assess the influence of environmental and biotic factors in the observed dispersal syndromes trends. Also, additional field studies on animal dispersers would give more insight on dispersal syndromes patterns found along tropical forests.
Adaptability of rare edge populations to climate change
Schwarzer, C 1 , Heinken, T 1 , Luthardt, V 2
1 University of Potsdam,
Research/Systematic Botany, Germany;
, Joshi, J 1
Biodiversity
2 University of
Applied Sciences, Eberswalde, Vegetational Science and
Applied Plant Ecology, Germany
The mires of Northern Germany are inhabited by several rare plant populations, which grow there at the southern margin of their species´ ranges. During the Pleistocenic glaciations, these cryophilous species were possibly widespread in Central Europe followed by range contractions to higher latitudes and altitudes during periods of global warming. Today, the few remaining populations in Northern Germany persist in isolated habitat fragments. According to the ‘Rear-edgehypothesis’, these populations experienced a comparatively long selection for tolerances of heat and drought stress, which could have resulted in specific local adaptations lacking in the Northern ‘core’ populations. Therefore, the role of these habitats for the past and future evolution of bog plant communities could be crucial as a possible shelter for well adapted populations to future climate changes. To test for differences in adaptive traits between core and edge populations, we planted populations of several bog plant species collected along a gradient from Northern Sweden to Northern Germany in a common garden into microcosms that (1) differed in species diversity and (2) were subject to different environmental conditions mimicking future climate projections (experimental drought in spring, artificially enhanced temperature in winter, enhanced nutrient supply). The expected data will allow predictions on prospective responses of wetland plant communities to climate change. These data may be important for the conservation of the few mires left in
Central Europe and for other populations that occur at the margin of their species´ ranges especially if the southern populations perform better in a drier and warmer climate compared to their northern conspecifics. In the first season, the dominant matrix species, Sphagnum magellanicum, grew better in mixtures than monocultures. This effect was especially pronounced in the most Northern populations that therefore benefitted most from species diversity.
Do large flowers have to be short-lived in stressful environments? Insights from the Cistaceae in a
Mediterranean ecosystem
Teixido, AL 1 , Méndez, M 1 , Valladares, F 2
1 Biodiversity And Conservation Area, Rey Juan Carlos
University, Mostoles, Spain; 2 Instituto De Recursos
Naturales, Consejo Superior De Investigaciones
Científicas, Spain
Larger flowers receive more pollinator visits increasing the reproductive success, but may also involve high maintenance costs associated with water use, especially in hot and dry ecosystems. In this context, short-lived flowers could play an important role to buffering such costs. We evaluated the factors influencing floral longevity in the Cistaceae, a large-flowered
Mediterranean family. Specifically, a study of floral longevity in response to differences in temperature, flower size and pollen deposition was conducted in the largest-flowered species, Cistus ladanifer . Floral longevity was recorded in 12 species covering diverse ecological ranges and differing in flower size. For C. ladanifer we conducted an observational study along an altitudinal gradient and an experimental manipulation of floral longevity. We pollinated and capped flowers along with flowers exposed to natural pollination. Different
GLMMs were used to evaluate both phenological and treatment effects on floral longevity. We included air temperature (ºC) and flower size as additional predictors to statistically control their potential effect on floral longevity. Flowers of Cistaceae tended to last one day,
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independently of differences in flower size amongspecies. However, our specific study of C. ladanifer reported some intraspecific variability in floral longevity, which was negatively related to flower size, pollen deposition and, especially, temperature. Flower size was significantly variable both among and withinpopulations, and also within-individuals; overall, larger flowers tended to be shorter-lived along the populations.
Relative to treatment effects, capped flowers lasted longer (1.56±0.55 days) that control (1.25±0.36 days) and hand-pollinated (1.04±0.11 days) flowers. Lastly, high temperature (approx. 20ºC) had a differential effect since limited floral longevity to one day independently of pollen deposition. Our results show a relationship between large and short-lived flowers in stressful environments such as the Mediterranean, and highlight the importance of floral longevity in this context. They also show that intraspecific variability in flower size as well as variation in temperature and disproportion in pollination can adjust floral longevity. We suggest that the adjustment of floral longevity in response to flower size and temperature is adaptative in large-flowered plants inhabiting Mediterranean ecosystems, where an adequate water use determines the floral maintenance costs under these conditions. The observed intraspecific variation in flower size could buffer the stress, minimizing these costs, and thereby, smaller flowers are persistent in populations.
Stomatal density of
Quercus pannosa
change over altitudinal gradient in Himalayas
Zhou, Z-K 1 , Xing, Y 2 , Su, T 2 , Huang, Y 2 , Hu, J 2
1 Kunming Institute of Botany, Chinese Academy of
2 Science, China; Xishuangbanna Tropical Botanical
Garden, CAS, China
The observation that changes in atmospheric CO
2
partial pressure have led to changes in stomatal density provided a proxy to reconstruct paleo-elevation in geological time by counting stomatal density from fossil leaves.
Quercus pannosa is a middle size tree which is widely distributed from 2500 to 4500 m in the Himalaya region. A lot of fossil leaves were found from the Neogene in the same region. Quercus pannosa has been identified as the nearest living relative of those fossils. Variations of stomata of this species among altitudinal gradient were reached in order to find the relationships between stomatal and venation density with CO
2 reconstruct paleo-CO
2 with altitude in density and
density. Stomatal density increase
Quercus pannosa from 565 to 752 /mm 2 when altitude increasing from 2500 to 4160 m. However, stomatal density is sharply decreased from 752 to
539/mm 2 , when altitude is over 4200 m. Fossil stomatal density of Quercus pannosa were counted. The CO
2 density in the Neogene and present-day can be compared.
Anthropogenic influence on atmospheric composition can be discussed. The project is support by National
Natural Science Foundation of China (41030212,
30970206)
The relationship between leaf margin characters and climate: a case study in China
Su, T 1,23 , Zhou, Z 1 , Jacques, FMB 4 , Xing, Y 5 , Huang, Y 5
70
1 Key Laboratory of Biodiversity and Biogeography,
Kunming Institute of Botany, Chinese Academy of
Sciences, Kunming, China; 2 Graduate University of the
Chinese Academy of Sciences, Beijing, China; 3 Current address: Key Laboratory of Tropical Forest Ecology,
Xishuangbanna Tropical Botanical Garden, CAS,
Mengla, China; 4 Dept of Palaeobotany and Palynology,
Nanjing Institute of Geology and Paleontology, CAS,
Nanjing, China; 5 Xishuangbanna Tropical Botanical
Garden, CAS, Mengla, China
The morphology of a leaf is largely affected by the surrounding environment. In this study, fifty samples from mesic to humid forests in China were chosen to understand the relationship between leaf margin characters and climate. the proportion of woody dicotyledonous species with untoothed leaves (P) shows a strong linear regression with mean annual temperature
(MAT), among the several currently proposed LMA equations, the one resulting from North and Central
American and Japanese data, yields the closest values to the actual MATs of the Chinese samples (mean absolute error = 1.9
˚ C). A new equation derived from the Chinese forests is therefore developed, where MAT ( ° C) = 1.038
+ 27.6 x P. Besides, the relationship between P and precipitation is found. P shows a high correlation to growing season precipitation (GSP, which is the total precipitation with mean month temperature more than
10 ° C). Whereas former works demonstrate that leaf size correlates most to precipitation parameters among leaf characters. A new equation is proposed, where GSP
(mm) = 200.9 + 1755.9 x P. This study improves the unique relationship between leaf margin characters and climate under the influence of monsoon over China.
1
A primary study on invasive plants in China
Ma, J 1 , Yan, X 1
Shanghai Chenshan Plant Science Research Center,
Chinese Academy of Sciences, China
Plant invasions have been attracting increasing attention worldwide because of their significant ecological impacts and economic costs. China is the world's third largest country with high plant diversity, spanning 50 degrees of latitude and five climatic zones, so it is vulnerable to the establishment of invasive species, especial alien species.
Therefore, it is essential for us to investigate the invasive species and their distribution area in China. In order to better understand their invasive characteristics, we intend to collect the primary data of these invasive or potential invasive plants in China by literature review and investigation, including the composition of plant taxa, their habits, invasive origin, introduced time, ways of introduction, most invasive habitats, invasive grades, and distribution in each province of China. A geographic and taxonomic overview of the invasive plants in China is given based on various reports in references. A total of
563 species have been recognized as invasive plants or potential invasive plant from China. The invasive plants are represented by 308 genera within 71 families, of which the Compositae, Gramineae and Leguminosae are

most important. Most invasive species in China originate from the Americas and Asia. Southeast coastal areas and south provinces of China, such as Guangdong, Hainan,
Fujian, have higher number of invasive plants than that in north areas. In order to evaluate the harmfulness, we divided these invasive species into 5 grades: (I) Most aggressive with large economic loss, like Eichhornia crassipes (Pontederiaceae), Eupatorium odoratum
(Compositae), etc; (II) Aggressive with ecologic and economic loss, but less than largely or broadly, like
Mikania micrantha (Compositae), Spartina alterniflora
(Gramineae), etc.; (III) Common weeds or unwelcome trees or shrubs, with damage less or not too evident, like
Solidago canadensis (Compositae), etc.; (IV) Locally aggressive, especially in Jilin, Beijing and Hong Kong, etc.; (V) Invasive occasionally, rarely, sparsely, scattered, or once invasive or widespread but more or less invasive, or not invasive yet but with potential ability in the future. The aim of our study is to provide basic information on the number and harmfulness of invasive plant species in China, and to seek more attention of the public for the increasing problems with invasive plants, besides further research work in the future.
How to conquer the desert? The neophytic flora of the
1
2
Southern Atacama Desert
Stoll, A 1,2 , Nickerl, J 3 , Squeo, F 4
Center of Advanced Studies in Arid Zones, Chile;
University of La Serena, Chile; 3 Dept for Botany,
Technical University Dresden, Germany; 4 Institute of
Ecology and Biodiversity
Neophytic invasions are one of the most challenging issues within conservation biology. Chile, like most of the other parts of the world, is faced with that problem with the particularity of being a biogeographical island.
Human activities, especially land use change, and globalization have become more intensive with a lasting effect on the Chilean flora. In our study we focus on the neophytic flora of 29 priority areas for biodiversity conservation defined in Atacama Region (Chile). Within these priority areas we registered 59 neophytes, mostly with a Holarctic origin (76%), but currently are cosmopolites. We reveal that neophyte plant species are spread over nearly 40% of the surface of the priority areas. More than half of this extent is occupied by neophytic annuals (61%), which are also the most represented life form (56%). In contrast native annuals only constitute 24% of the flora in Atacama Region.
Parallels between the patterns of native and neophytic flora are detected in terms of the most diverse families
(Asteraceae and Poaceae) and altitudinal distribution of species diversity (high in the Coastal sector). A comparison with similar regions in the world reveals highest similarities with the neophytic flora of Argentina and Australia. The main invaders of different native ecosystems are Erodium cicutarium and
Mesembryanthemum crystallinum , special attention is laid on worldwide invaders as Arundo donax and
Tamarix ramosissima . Meanwhile most species still not present invasion characteristics, they represent a potential risk for the conservation efforts of the threatened native flora and natural ecosystems of the Southern Atacama
Desert.
Catchment scale dispersal of the invasive willow,
Salix cinerea
, in south-eastern Australia
1
Hopley, T 1,2 , Young, AG 2 , Zwart, A 2
Research School of Biology, Australian National
University, Canberra, Australia; 2 CSIRO Plant Industry,
Canberra, Australia
Willows are aggressive exotic components of many river systems in south-eastern Australia and they have the potential to expand their range. Current control efforts for the most highly invasive willow species, Salix cinerea , are extensive, costly and not always successful due to rapid post-removal reinfestation. An improved knowledge of the reproductive ecology and dispersal dynamics of this species will help to minimise future expansion and make current control efforts more effective. The species was found to be predominately insect pollinated but can exhibit wind pollination which has the potential for long distance dispersal. Seed production is highly variable across the catchment with average seed production over 300,000 seed per tree, equating to over 150 million seed produced in just 12 km of infested river. The sites with the highest density of larger trees are producing the most seed. A disproportionally high reduction in propagule pressure may be attained by first clearing those sites with the highest contributions. Clearing just 20% of the sites sampled would reduce seed production in the catchment by over 50%. This high reproductive output combined with high initial germination rates, above 95%, gives the species the potential to reinfest cleared areas and expand its range. A survey of populations in the Ovens River catchment of south-eastern Australia was undertaken to determine seed and pollen movement within and between populations of S. cinerea . Paternity analysis using molecular markers show that nearly 40% of seed on trees are sired from outside the home location. Genetic profiling of populations in surrounding rivers has allowed us to identify the most likely pollen and seed sources to give an estimate of the scale of gene flow.
Results suggest that at least 40% of pollen is moving more than 15 km between rivers. Seed dispersal shows similar patterns with 50% of seed dispersal at more than
15 km and a small amount travelling over 30 km. These results suggest that land managers need to co-ordinate control efforts across rivers for more effective long term eradication.
Impacts of the invasive vine
Merremia peltata
on the rainforest tree
Dillenia biflora
and its bat pollinators in Fiji
Scanlon, A 1 , Petit, T 1
1 University of South Australia, Adelaide, Australia
The invasive vine Merremia peltata (Convolvulaceae) blankets tree canopies in the rainforest and cultivated areas of Fiji, but its effects on forest processes is unknown. We monitored flower and fruit production in two groups of the common rainforest tree Dillenia biflora (Dilleniaceae), a bat-pollinated species. Five trees were cleared of the vine and five remained covered with it. We counted buds, flowers, and fruits each month for
19 months (2009–2011). Evidence of animal visits to
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flowers (bite marks and infrared camera footage) was recorded. Trees with no M. peltata produced significantly more buds, flowers, and fruits than did covered trees, but flower abortion rates did not differ between the two treatments. The proportion of D. biflora flowers that were pollinated under M. peltata was much smaller than that of flowers without the M. peltata blanket, indicating that the vine is decreasing access to the bat pollinators.
The study shows that M. peltata decreases forest productivity, including resources for bats. M. peltata should be controlled in protected rainforest reserves.
The adaptive capacity of native and exotic grasses under climate change
Godfree, RC 1
1 CSIRO Plant Industry, Canberra, Australia
Over the coming century, an increase in the frequency and severity of extreme climatic events is expected to result in global shifts in vegetation structure, composition and distribution, as plant populations increasingly face abiotic stresses that lie beyond their physiological tolerances. However, in most cases we know little about the specific traits that are likely to be placed under selection pressure, the magnitude and distribution of population- and species-level variation in these traits, and the relative roles of demography, evolution and plasticity for coping with increased abiotic stress. Here I present preliminary results from a study aimed at determining the adaptive capacity of populations of two perennial, mainly inbreeding stipoid grass species, Austrostipa bigeniculata , which is native to Australia, and the invasive exotic Nassella neesiana , to future climate change. Four populations from both species sourced from a dominant climatic gradient were assessed for variation in climate-linked physiological and morphological traits, and then, using a new experimental system for simulating climate change in the field, subjected to a range of abiotic stress levels. The results suggest that interpopulation variation for most climate-linked physiological and morphological traits is higher for N. neesiana than for A. bigeniculata , but that withinpopulation genetic variation and heritability is low in both species. This suggests that populations of many dominant native and exotic Australian grasses may have limited capacity to respond to climate change via selection for climate-related traits.
Biotechnology and ex situ conservation: the ‘last resort’ for critically endangered plants in biodiverse regions
Bunn, E 1,2 , Turner, S 2 , Dixon, K 2
1
2
Botanic Gardens And Parks Authority, Perth, Australia;
2Faculty of Natural and Agricultural Sciences, School of Plant Biology, University of Western Australia,
Nedlands, Australia
Implicit in ‘Priority for change 1: Building ecosystem resilience’ in Australia’s Biodiversity Conservation
Strategy 2010–2020 document is the need to ‘Maintain and enhance ex situ conservation measures as a last resort for biodiversity conservation’. Biotechnology has a key role to play in the form of molecular, in vitro and cryogenic research that provides ‘emergency treatment’ for saving endangered species threatened with imminent extinction. Selected provenances that can ensure sufficient genetic representation for restoration can be identified by genetic fingerprinting technology. In vitro research can be utilized to micropropagate scarce plant material (and where seed may not be available), then in vitro culture lines of key provenances can be maintained ex situ permanently in culture collections and/or cryopreserved in liquid nitrogen until required for propagation and restoration of declining populations or reintroduction of species that may have become extinct in the wild. Cryopreserved material can be efficiently stored than revived and utilized to ensure remedial stocking of restoration sites to boost genetic representation as required. Monitoring and further genetic sampling can provide early warning of inbreeding depression among offspring in repatriated populations. Examples of biotechnology research for conserving critically endangered plants in biodiverse regions (such as the
SWAFR) are presented.
Predicting success in the reintroduction of the endangered Hawaiian fern
Marsilea villosa
: an
1 experimental case study
Chau, M 1 , Ranker, T 1
University of Hawaii at Manoa, USA
Conservation of rare and endangered plants can be accomplished by the practice of outplanting, but ecological management is often required for new populations to become established. In species with narrow habitat requirements, it can be difficult to predict which management methods will be successful at new locations. Marsilea villosa (‘ihi‘ihi) is an endangered
Hawaiian fern with only seven populations, all in ephemerally flooding drylands. Among its unique traits are long-lived sporocarps (i.e., highly modified leaves containing sporangia and spores), a requirement of flood and drought to complete its sexual life cycle, and vegetative growth in absence of flood. Much of Hawai‘i experienced an extended drought from summer 2009 to winter 2010, prompting concern for many endangered plants. Despite ‘ihi‘ihi’s high drought-tolerance, its cover at three O‘ahu populations decreased from January 2009 to January 2010 (peaks of rainy seasons). A common garden experiment was performed to address the question: Which restoration management techniques will facilitate growth of outplanted M. villosa ? The following effects were tested in a split-plot factorial design: two flooding levels (once/none), two light levels (50% shade/full sun), two weeding levels (bi-monthly/none), and the interactions of these factors. Clumps of ‘ihi‘ihi rhizomes were collected from a natural population at
Lualualei, O‘ahu and grown in pots, and 48 similar plants were randomly transplanted into experimental plots, in soil also collected from Lualualei (containing a weed seed bank). Percent cover was measured for M. villosa and analyzed using multiple-factor ANOVA to determine
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how much it expanded in response to treatment combinations. Sporocarps were counted at the end of the
12-week experiment. Results showed that, as expected, flooding had the most significant single-factor effect on increased M. villosa cover (P=0.001) and sporocarp production (P=0.002). However, shade also increased cover over the full sun treatment (P=0.004) when the plants began to experience drought at the end of the experiment. There was an interaction effect of light*flooding (P=0.004) because M. villosa grew best in flooding, shaded plots. Weeding had no significant effect throughout the experiment. Beyond protected status, current in situ management of ‘ihi‘ihi populations consists entirely of weed management, by mowing or herbicide. While managers hope that extant populations can recover with subsequent flooding, there is concern that the dry environments where M. villosa currently exists may only become drier with climate change.
‘Ihi‘ihi was historically found in a few moderately wetter valleys on O‘ahu, but those populations were lost to land development. The results of this study show that laborintensive weed management may be unnecessary if reintroduced M. villosa is planted under conditions of flooding and moderate shade, and if planted at the start of a rainy season, will require minimal management to become a self-sustaining new population.
Sussex communities: plants and people
Sutcliffe, J 1
1 Manhood Wildlife And Heritage Group, West Sussex,
UK
The importance of volunteers and the need for local communities to be involved in protecting their local wildlife is enshrined in the Convention on Biological
Diversity (1992) and, in turn, by national governments such as that of the United Kingdom. In 1997, an innovative project was established by local people in
Selsey, West Sussex, UK. Their work is presented as a case study. This pilot initiative has been extended to involve all 11 parishes on the Manhood (derived from
Main Wood) Peninsula which lies south of Chichester. In
2004, the creation of a Biodiversity Duty for public bodies and an increasing workload which exceeded the time available to the volunteers led to the group appointing a part-time member of staff and a new working relationship with Chichester District Council.
2010 was the United Nations International Year of
Biodiversity. In June, the group was awarded the
Queen’s Commendation for Voluntary work – a fitting tribute for some 200 volunteers who have contributed over 11,000 hours to conservation.
Landscape structure and pollinator behaviour do not predict reproductive connectivity among populations of the bird-pollinated shrub
Eremophila glabra
Elliott, CP 1,3
S 4 , Young, A 1
, Zwart, A 2 , Lindenmayer, D 3 , Cunningham,
1 CSIRO Plant Industry, Canberra, Australia; 2 CSIRO
Mathematics, Informatics and Statistics, Canberra,
Australia; 3 Fenner School of Environment and Society,
ANU, Canberra, Australia; 4 CSIRO Entomology,
Canberra, Australia
The maintenance of viable populations in heavily fragmented landscapes is thought to be mediated by connectivity among populations. Connectivity can be measured structurally (spatial configuration of the landscape) and functionally (behavioural responses of organism's to fragments and landscape structure). Given this, understanding how structural connectivity translates to functional connectivity is important, especially from a management perspective, because such landscapes are often managed structurally but it is the functional connectivity that is the desired conservation management outcome. To investigate this relationship we assessed connectivity in fragmented landscapes in south-eastern
Australia, by examining plant-pollinator interactions of a common, bird-pollinated, autohexaploid shrub
( Eremophila glabra ssp. glabra ). We contrasted populations occupying three replicated landscape contexts that ranged from large intact vegetation patches to small isolated linear strips. We compared the response of pollinator communities and plant reproduction to these conditions, and measured functional connectivity by linking the movement of pollinators and pollen together.
Reduced structural connectivity resulted in significantly different pollinator communities and lower seed set in the small linear strips compared to large intact patches.
However, two lines of evidence suggested this did not translate into reduced functional connectivity. First, pollinator behaviour of the different communities was similar among the different populations, and second, all had similarly high levels of pollen immigration as assessed by paternity analysis. We attribute this maintenance of high functional connectivity to notably mobile pollinators and a self-incompatibility system that enhanced the genetic connectivity among populations by favouring foreign pollen. Therefore, we conclude that reduced structural connectivity did not result in equivalent loss of functional connectivity, and pollinators were key to maintaining the current immigrant gene flow crucial for these highly fragmented plant populations.
Floral biodiversity in New Guinea: how many species are there?
James, S 1 , Allison, A 1 , Snow, N 1
1 Bishop Museum, Honolulu, USA
New Guinea, the world’s largest (~890,000 km 2 ) and highest (4,980 m) tropical island, is considered one of the most species rich areas on Earth, hosting approximately
6% of the world’s flora and a high level of species endemism. However, there remains considerable speculation as to the size of the vascular flora, with published estimates ranging from 11,000 to 25,000 species. An ongoing systematic and collaborative effort to develop a species checklist from published literature, online resources, taxonomic experts, and cross-correlated with specimen vouchers, suggests that the number of currently accepted native vascular plant species is about
13,500, with almost a quarter of the species comprising the extremely diverse orchid family. Because New
Guinea is one of the most poorly collected regions of
Melanesia, localities predicted to be species rich and currently under-collected are actively being surveyed.
The databasing of herbarium label data, georeferencing of locality information, imaging of specimens, and making this data available online, is aiding the
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clarification and study of the biodiversity and distribution of the vascular flora of New Guinea. Such uncertainty in the knowledge of the taxonomy and extent of botanical diversity of the region hampers both the prediction of the effects of anthropogenic actions, such as forestry, mining, and climate change, and the effective development of conservation plans. With forest across
New Guinea being removed or degraded at the rate of at least 1.4% per year, the number of species threatened with extinction is, no doubt, greater than the 1% currently listed by the IUCN.
Understanding New Guinea botany: collecting density and floristic patterns
De Kok, R 1 , Utteridge, T 1 , Briggs, M 1
1 Royal Botanic Gardens, Kew, UK
The large tropical island of New Guinea is considered one of the world's last remaining wilderness areas with extensive areas of intact lowland forest surrounding a mountainous central range. Botanical exploration on the island has a long history closely linked to European settlement in SE Asia and Australia and the associated scientific development in the region. We will briefly discuss these historical collecting programmes and the resulting legacy of botanical research available for contemporary researchers. Using data from historic herbarium collections as well as data from modern general collecting floristic programmes, we will examine collecting patterns on the island. Through this we hope to highlight particular habitat types and areas of the island that can be considered over- and under-collected. We will discuss these patterns in light of conservation needs, and hope that this paper will provide the botanical, as well as the wider science and conservation communities, with a direction for future work on the island.
Seed ecology of threatened species: implications for long-term management
Turner, S 1 , Bunn, E 2 , Dixon, B 2 , Merritt, D 2 , Dixon, K 2
1
2
University of Western Australia, Perth, Australia;
Kings Park and Botanic Garden, Perth, Australia
For the long-term conservation of threatened flora there are many factors that need to be investigated to ensure the maximum likelihood that a species will continue to exist in situ. A number of these factors are well recognised including conservation genetics, ecology and the factors that are likely to influence in situ reintroduction. The seed ecology of threatened species is one of those often overlooked research areas that may take longer to yield results (potentially many years) but nevertheless is of significant importance as it will allow current and future managers to predict the likely longevity of the soil seed bank or even provide information as to the likelihood that a species that is locally extinct may still persist in the soil seed bank and what factors are likely to stimulate its germination after
74 many years of absence. Since the beginning of 2009 the seed biology and seed ecology of two critically endangered species, Commersonia sp. Mt Groper
(Malvaceae) and Symonanthus bancroftii (Solanaceae), has been investigated. Commersonia sp. Mt Groper has fewer than 80 plants remaining in the wild and is only known from three small populations from the Albany region near the south coast of WA. In comparison,
Symonanthus bancroftii is a small dioecious shrub known from only one location near Bruce Rock in the WA wheat belt, where just two plants were discovered in remnant bushland in 1996 and 1998. Seed from both species when fresh were found to be highly dormant – the seeds of C . Mt Groper were found to possess physical dormancy while seeds of S. bancroftii were found to have physiological dormancy. Subsequent research has since found that C . Mt Groper seeds readily germinate when they are either nicked or exposed to hot water prior to incubation, while fresh seeds of S. bancroftii only germinate when precision nicked and treated with 100 ppm GA (81%) – other treatments assessed (i.e. Kar1 and dry heat (100°C for 10 mins)) proved to be completely ineffective. The use of nicking and GA for S. bancroftii while a good way to stimulate germination under laboratory conditions nevertheless does not shed any light on the drivers of dormancy loss under natural conditions. Subsequent experimentation has since found that S. bancroftii seeds become Kar1 (smoke) responsive during after-ripening though the rate of after-ripening is relatively slow with only 41 % germination after 12 months dry storage. A seed burial trial (still underway) has also confirmed that seeds maintain high viability
(90%) during soil storage over the first season and also become Kar1 responsive. Base on these preliminary results it appears that S. bancroftii may be a fireresponsive species only recruiting following a bushfire after a period of after-ripening in the soil seed bank, while Commersonia Mt Groper seeds in comparison have physical dormancy and as with other species with this form of dormancy are likely to recruit following some type of soil disturbance as well.
Management rather than climate explains demographic variation in
Caladenia orientalis
, a threatened orchid from south-eastern Australia
, Duncan, MJ 1
1
Coates, F 1
Dept of Sustainability and Environment, Heidelberg,
Victoria, Australia
Caladenia orientalis (G.W.Carr) Hopper & A.P.Br. is a critically endangered orchid. The largest known populations are confined to fire-managed coastal heathland in southern Victoria. Trends in population dynamics at two closely occurring sites were evaluated against time since fire and rainfall to provide ecological and biological information relevant to population management. At both sites, decreased plant size was inversely correlated with time since fire and the number of non-reproductive plants was positively correlated with time since fire. Rates of flowering were inversely correlated with time since fire at only one site. The vegetation at this site rapidly accumulated after fire, whereas recovery was relatively slow at the other site.
Rainfall was not correlated with rates of flowering or leaf width at either of the study sites, although there was a

weak inverse relationship between rainfall and the number of non-reproductive plants at one site. Rates of pollen transport and fruit set were within reported ranges for deceptive species. Fruiting plants were significantly smaller in the following year, whereas non-reproductive plants remained the same size. The results suggest that there may be substantial costs associated with reproductive effort, and that hand-pollinating plants to boost seed production may lead to decreased plant size in the following year. Annual variation in rates of flowering may be influenced by previous reproductive effort.
However, long-term population trends are better explained by competition from dominant shrubs, which become increasingly abundant with a lack of fire. Both populations are fire-managed according to prescriptions generally considered necessary to maintain the structure and floristics of heathland although sites differ markedly in their topographic position. This difference may underpin the differences in behaviour between the two populations and may provide insights for site management. Although increased drought is predicted in south-eastern Australia as a consequence of global warming, implementation of appropriate fire regimes is likely to be a more immediate issue than rainfall at our study site. The use of fire to manage C. orientalis habitat, prevent over-abundance or further encroachment of the dominant shrub Leptospermum laevigatum and preserve broader biodiversity values is likely to be a prime challenge for preventing the decline of this critically endangered orchid. However, these relatively large C. orientalis populations occur close to land now highly sought after for development and there is increasing concern that prescribed fires might escape control lines and damage adjacent properties. Our results imply that alternative methods to boost fecundity of selected plants, such as hand-pollination, may actually compromise population fecundity. Developing site-specific fire regimes to promote better habitat is the way forward.
Demographic studies result into adapted management strategies for the rare
Helosciadium repens
(Apiaceae)
Ronse, A 1
1 National Botanic Garden of Belgium, Meise, Belgium
Helosciadium repens (Jacq.) W.D.J.Koch, an umbellifer formerly known as Apium repens (Jacq.) Lag., is a clonal plant species that is rare and endangered throughout its natural distribution area (mainly Atlantic northwest
Europe). It is mentioned in annex II and annex IV of the
Habitat Directive of the European Union (Directive
92/43/EEG), listing the species of community interest in need of strict protection. In Belgium, that lies in the central part of its distribution area, it is a very rare species and only five populations exsist. Aiming at developing a conservation strategy for this species, a research programme on its conservation biology was carried out. In a preparatory phase European populations of Helosciadum repens were screened on their habitat ecology and vegetation characteristics, and phytosociological releves were made. From 2003 to 2007 demographic monitoring measurements were carried out six times a year in twenty permanent plots in four
Belgian populations. Germination, flowering, seed setting and clonal growth were recorded in the plots. At the same time several parameters relating to grazing or other management regimes were recorded, such as vegetation height and openness of the vegetation, as well as groundwater levels near the plots. Climatological data were obtained from official meteorological stations in the vicinity. Soil samples were taken in order to investigate the nutrient requirement of the species. Additionally, the soil seed bank of two populations was analysed by means of a long lasting germination experiment. Our field observations and demographic results have given a good insight in the life history characteristics of this unsufficiently known species, as well as in the threats that weight upon it. A life cycle graph was developed, making a distinction between main ramets, grown up from seedlings, and clonally formed side ramets of different orders, thus quantifying both the clonal and the sexual reproduction. The plants have a perennial life cycle, individual ramets being able to survive for several years in favourable conditions. They form a soil seed bank with a germination that is delayed over several years. Analyses of population matrices display different growth rates in different localities and periods. There have been dramatic decreases in three of the four populations during our investigation period. Population growth rates are related to the applied management regimes, showing this to be a key factor for survival of this species. Nonetheless, large differences in number of ramets were also found, that were only attributable to climatological parameters, mainly drought. We found a significant relationship between the number of ramets, the groundwater level and some management parameters, such as vegetation height. The flowering and seed setting were influenced by climatological parameters. Overall, it appears that the main threat for this endangered species is the loss of suited habitat, and that there has been a shift in type of habitat.
Conservation challenges on Mediterranean
1 gypsophytes
Moreno Saiz, JC
Fuster, MM 3
1 , Fernández-Mazuecos, M 2
, Pérez García, N 4
, Génova
, Domínguez Lozano, F 5
Dept Biología (Botánica), Universidad Autónoma de
Madrid, Spain;
Spain; 3
2 Real Jardín Botánico de Madrid, CSIC,
Dept Producción Vegetal, Universidad
Politécnica de Madrid, Spain; 4 Dept Biología Vegetal,
Universidad de Barcelona, Spain; 5 Dept Biología
Vegetal I, Universidad Complutense de Madrid, Spain
Semiarid environments over gypsum soils occupy a large area in the Mediterranean basin and, accordingly, in the
Iberian Peninsula as well (c. 150,000 ha). The flora of peninsular Spain includes no less than 140 gypsophytes
(J Mota et al. 2009) usually occurring in open grasslands and scrublands within a matrix of heavily humanmodified landscapes. Among the most characteristic taxa of those communities is the sub-shrub Vella pseudocytisus L. (Brassicaceae), including two endemic subspecies in Spain. Their distribution areas are heavily fragmented and placed c. 200 km apart. Both subspecies have been included as Endangered in the national Red
List (J.C. Moreno. 2008) and protected by regional laws in Madrid and Aragón. Since 2001, our research group performs a demographic monitoring of Vella pseudocytisus using matrix models to analyze population dynamics and future trends in vital rates. Seedlings experience mortality rates of 90%, but mortality drops
75

dramatically as individuals reach the adult stages. Adult survivorship is key for population viability, with changes in vital rates at that stage having a strong impact in overall population dynamics. Fortunately, a dendrochronological study in course shows that the species is a relatively long-lived shrub with their eldest individuals achieving nearly 50 years. Although some between-population variability in demographic rates has been assessed in our studies, 30 to 100 years projections performed for all populations result in a negative growth rate. Such observed population trends are discussed in relation to other studies performed on threatened Spanish or Mediterranean gypsophytes and xerophytes. Global and land-use changes will play a crucial role in the conservation of these species and their communities.
Creating rare from common species – seed sourcing and restoration of iconic yellow box (
Eucalyptus melliodora
)
1
Broadhurst, L 1
CSIRO Plant Industry, Canberra, Australia
Yellow box ( Eucalyptus melliodora ) is a valuable and iconic tree once broadly distributed from western
Victoria, New South Wales and southeastern
Queensland. The soils coinciding with this distribution are favourable to cropping, horticulture and grazing which has resulted in severe loss and fragmentation of this species and the ecological community in which it resides. This ecological community is now protected under both Commonwealth (EPBC) and State (NSW) legislation. Yellow box is valued for its shade, shelter and timber qualities as well as being important for wildlife habitat and honey production. As such it is important restoration species throughout its distribution.
However, given the high levels of fragmentation that this species has undergone it is likely that low genetic diversity and elevated inbreeding may limit the utility of some populations to act as restoration quality seed sources. Seed collected under current seed sourcing protocols from across the southern tablelands of NSW were assessed for levels of genetic diversity to help improve yellow box restoration within this region.
Excess seedlings from this analysis were later planted into two provenance trails and similarly assessed to determine how much diversity was captured during this process. Finally, mating patterns and pollen dispersal were assessed in five reproductively mature restored sites to determine whether these would be suitable seed sources for future restoration projects.
Plant breeding systems in alpine ecosystems in the southern South American Andes – searching for signs of vulnerability under climate change
Arroyo, MTK 1 , Humaña, AM 1
1 Instituto de Ecología y Biodiversidad, Facultad de
Ciencias, Universidad de Chile, Santiago, Chile
Alpine plants may cope with climate change by upward migration along steep altitudinal gradients. The ability of plants to keep up with the pace of climate change will be influenced by dispersal capacity and breeding system.
Among species with different breeding system, those
76 with strong autonomous selfing capacity should be best adapted for migration along altitudinal gradients on account of their capacity to produce seed without the intervention of biotic pollinators. At the other extreme, successful migration of self-incompatible and sexually dimorphic species with specialized pollination might be impeded if pollinators and plants migrate upward at different rates, as would be expected from their respective life history attributes. The autonomous selfing hypothesis posits that generally cold temperatures and variable weather conditions at high elevations will select for autonomous selfing in high elevation ecosystems, which might suggest that alpine plants are well adapted to cope with climate change. Here using data from a published community study for the Patagonian alpine
(50°S) in southern Chile and a parallel ongoing study in the central Chilean Andes (33°S) (together >200 species) we investigate the incidence of breeding systems along the self-incompatibility (SI)-self-compatibility (SC) gradient. Considering the dominant perennial herb lifeform, where good breeding systems records are most abundant, both alpine sites show a wide range of breeding systems, there being large contingents of selfincompatible and partially self-compatible species habitually serviced by biotic pollinators, in addition to strongly autogamous species. Contrary to expectation, the frequencies of SC and SI species in the warm and sunny central Chilean alpine and the cold and windy
Patagonian alpine are similar, despite wide communitylevel differences in flower visitation rates. Large breeding system data sets are scarce worldwide. The available information suggests that in spite of the cold and variable weather conditions at higher elevations, SI in perennial herbs is as well represented in the high South
American Andes as in some lowland communities, and that many alpine species in this region of the world could indeed be vulnerable under climate change. Research financed by Fondecyt Grant 1085013 and IEB grants,
Chile.
Plant architectural and genetic diversities in
Coffea native from Madagascar: towards an architecturalfunctional plant growth model applied to
Coffea biodiversity perservation
Sabatier, S 1 , Andrianasolo, D 1
, de Reffye, P 1
, Rakotomalala, J-J 2
, Letort, V 4
,
1
Hamon, P 3
4
CIRAD, UMR AMAP, Montpellier, France;
Antananarivo, Madagascar;
2 FOFIFA,
3 IRD, Montpellier, France;
ECP, Paris, France
The Coffea genus includes 103 species with 53 endemic to Madagascar (namely Mascarocoffea).The three main centres of species diversity for coffee trees are
Madagascar (naturally low-caffeine species), Cameroon and Tanzania. However, Madagascar has the higher
Coffea diversity in terms of total number of species due to the great variety of forest types including littoral, evergreen, gallery, mixed deciduous, dry, xerophytic and high-altitude. During these last 20 years, deforestation in
Madagascar has become dramatic and has drastically reduced the distribution area of Coffea species. The collection maintained at the Kianjavato Research Station is unique since no living material is authorized to be conserved out of Madagascar. Conservation strategies based on scientific data are needed and become urgent in

terms of biodiversity preservation and sustainable development. Recent studies in Coffea trees showed that their phenotypic plasticity could be explain the restricted distribution of C offea species. Our recent preliminary results showed a high architectural diversity (i.e. architectural model, growth process, branching degree, branch location) between Coffea species and an interspecific variability of functional traits (Leaf Area
Mass, wood density). The functional-structural model
GreenLab is a dynamic model taking into account architectural plasticity of the plants and biomass allocation to organ level. Potentials of GreenLab model to represent environment x genotype interactions, in particular through its stable endogenous parameter as such the organ sink functions have been demonstrated in some species and they will be tested for Coffea species.
The present study focuses on 5 species endemic to
Madagascar with different phenotypic traits and forest habitats. For each species, architecture and genetic comparative analyses between individuals growing in situ (natural forest) and ex situ (common garden test) will be performed. In addition, we have set up two experimental stands to analyse the site effect. These whole results will be used to introduce genetic factors into the GreenLab model. The global aim of our approach is to analyse the adaptive capacity and phenotypic plasticity at the individual level within Coffea populations.
4
3
C
4
Rice Project
1
Quick, WP 1
International Rice Research Institute, Philippines
Future global population increases of some 2 billion people by 2050 will require an extra 250 million tonnes of rice per year in Asia alone. To feed this increased population, agriculture will need to improve food production by some 50% equivalent to a second green revolution that first occurred in the 60's and 70's. The C
4
Rice Project was launched 18 months ago to address this problem and will need substantial financial investment now and in the future if it is to succeed. Excellent initial progress has been made in the project which is predicted take 15–20 years. This talk will provide a description of the consortium, outline the research strategies being employed, give an update on current research progress and outline some of the challenges to be tackled in the coming years.
1
Engineering the C
4
pathway into rice
Hibberd, J 1 , Covshoff, S 1 , Woodfield, H
Slamet-Loedin, I 2 , Quick, P 2
University of Cambridge, UK; 2
1 , Karki, S
IRRI, Philippines
2 ,
It has been proposed that integrating C
4 into C
3
photosynthesis
crops could be used to increase their maximum yields. This is a challenging undertaking because the C
4 pathway is complex and involves alterations to the biochemistry, cell biology and development of leaves.
However, C photosynthesis has evolved independently
4 at least 62 times, and this biological precedent implies that changes to a relatively small number of factors may generate the complexity of a C
4
leaf. Recent advances in proteomics, transcript profiling and the release of genome sequences for sorghum and maize, two important
C crops, have filled some of the gaps in our
4 understanding of mechanisms underlying C
4 photosynthesis. Further advances are being made by comparative analysis of closely related C
3
and C
4
plants because this reduces signal noise associated with phylogenetic distance. We will consider both our understanding of global alterations to gene expression in
C
4
compared with C
3
leaves, and also provide specific examples of how the regulation of genes has altered as they are recruited into the C
4
pathway. We will discuss evidence that distinct lineages of C
4
plants share regulatory circuitry that generates accumulation of photosynthesis proteins in specific cells. A summary and progress report of current efforts at placing the biochemistry of C provided.
4
photosynthesis into rice will be
1
Evidence for the essential components of the C
4 mechanism
Furbank, R 1 , Pengelly, J
Voznesenskaya, E
Von Caemmerer, S
4
2
2
, Grof, C
, Martin, A 3
3 , Byrt, C 3
, Evans, J
, Edwards, G
2
5
High Resolution Plant Phenomics Centre, CSIRO Plant
Industry, Canberra, Australia; 2 Research School of
,
,
Biology, The Australian National University, Canberra,
Australia;
Australia; 4
3 University of Newcastle, Newcastle,
VL Komarov Botanical Institute, St
Petersburg, Russia; 5 School of Biological Sciences,
Washington State University, Pullman, USA
Recently there has been a renewed interest in the C
4 photosynthetic mechanism due to the desire to improve yield potential and radiation use efficiency in C
3
crops by introducing elements of the C
4
pathway (C
4
Rice
Consortium; http://beta.irri.org/projects15/c4rice). To facilitate this process, strategies need to be developed to identify the subset of key genes necessary to transfer a functional C
4
-like mechanism to C
3
crops. This presentation describes development of phenotypic screens for ‘C
4 transgenic C
3
-ness’ which can be used in analysis of
plants in which partial C
4
pathway components are being installed, analysis of photosynthesis in sorghum with impaired lignification and secondary thickening of leaf tissues, and photosynthetic characterisation of maize husk, and leaf sheath, tissue thought to operate a biochemical and structural blend of C
4
and C
3
photosynthesis. These data
77

are synthesised to delineate the challenges of measuring
‘C
4
-ness’, the role of bundle sheath cell wall properties in the efficiency of the C
4
CO
2
concentrating mechanism and the key steps in attaining a C
4
-like C
3
crop plant.
Do rice mutant populations offer variation in traits useful for C
4
engineering ?
Murchie, E 1 , Quick, P
, Feldman, A 1
2 , Leung, H 2 , Ahamadeen Nagoor,
1
MM 1
University of Nottingham, School of Biosciences, Div.
Plant and Crop Sciences, UK; 2 International Rice
Research Institute, Los Banos, Philippines
A large proportion of global rice production takes place in conditions where high temperatures induce substantial levels of photorespiration resulting in deterioration of radiation use efficiency. The introduction of the C
4 pathway would be of substantial benefit under these conditions and provides a solution for the attainment of
2050 yield targets. The rice genome may contain sufficient ‘plasticity’ to enable the introduction, or preconditioning of some traits that would be essential for the engineering of the C
4
pathway in rice. The C
4
rice consortium at the International Rice Research Institute
(IRRI) in the Philippines has begun a high throughput screening program for available rice mutant collections including both irradiation/chemical mutagenesis lines and tagged lines. To date around 12,000 M4 lines from the existing IR64 deletion mutant collection have been screened at IRRI for vein density and 5,000 have been screened for tolerance to low CO
2
. Ten progeny of each line were visually scored for vein density and then screened in a chamber for 6 days at 30
µ
L L -1 CO
2
. High vein density candidates were further screened for leaf structure including inter-veinal mesophyll cell number and size. Initial results indicate a detection frequency of
1–2 candidates per 1,000 lines with both increased vein density and reduced mesophyll cell number. Vein density was stable during different growth stages in the field.
After 6 days at low [CO ] mutant lines were identified
2 that had a similar loss of chlorophyll to the C
3
–C
4 intermediate Panicum milioides control which suffered only marginal loss. We are testing the hypothesis that chlorophyll loss is a sufficiently robust and convenient proxy to use in high-throughput screens. A continuous program of phenotype confirmation, backcrossing and genetic analysis is underway in order to seamlessly incorporate promising traits into a breeding program.
‘State of play’ results will be described and discussed.
The roles of C
4
Leegood, R 1
acid decarboxylases in plants
1 University of Sheffield, UK
Any attempt to engineer C
4
photosynthesis into crop plants will require the recruitment of a decarboxylase in the form of NAD- or NADP-malic enzyme, or PEP carboxykinase (PEPCK). NAD-malic enzyme is probably ubiquitous in plant cells (although it may be enriched in some of them), but both NADP-malic and PEPCK show discrete locations. For example, NADP-ME is present in the trichomes, hydathodes, vasculature (cytosolic) and guard cells. PEPCK is present in many plant tissues and
78 appears to be located exclusively in the cytosol, although it may be plastidic in diatoms. Within leaves, although not abundant overall, PEPCK may be present in phloem companion cells, hydathodes, trichomes and stomata and its abundance is often dependent on developmental stage.
PEPCK is regulated in some, but not all, plant tissues by reversible protein phosphorylation, whereas the regulation of NADP-malic enzyme lacks such mechanisms and the regulation of NAD-malic enzyme remains unclear. In recent years it has become apparent that PEPCK functions in far more metabolic processes in plants than was previously realised, but, as in many animal tissues, its function has been less easy to discern.
It has been identified in a range of tissues, such as developing seeds, flowers, roots, vascular tissue and fruit. In developing seeds laying down storage proteins it is implicated in the metabolism of incoming nitrogenous compounds, as it also is in the phloem. In stomata, new data from Arabidopsis knock-outs suggest that it plays an important role in stomatal closure, a role that may also be played by NADP-malic enzyme.
Is corn husk photosynthesis C
4
Pengelly, J
-like?
1 , Evans, J 1 , Voznesenskaya, E 3
1
, Edwards, G 4 ,
1
Furbank, R 2 , Von Caemmerer, S
Research School of Biology, The Australian National
University, Canberra, Australia; 2 High Resolution Plant
Phenomics Centre, CSIRO Plant Industry, Canberra,
Australia; 3 VL Komarov Botanical Institute, St
Petersburg, Russia; 4 School of Biological Sciences,
Washington State University, Pullman, USA
The husk tissue surrounding the ear of Zea mays
(maize/corn) has been described as operating a partial C
3 photosynthetic pathway in contrast to its foliar leaves which function using the NADP-ME C
4
pathway for CO
2 assimilation. Here we characterized photosynthesis in maize husk and leaf, measuring the defining C
3
/C
4 characteristics including vein density, combined gas exchange and carbon isotope discrimination, the O dependence of the CO
2
2
-compensation point, and photosynthetic enzyme localisation. Vein density in husk tissue was 88% less and an altered stomatal number and distribution were observed in relation to leaf tissue. The
CO than in leaves and did not saturate at high CO
CO
2
2
response of CO
2
assimilation rate in husk was less
2
indicating
diffusion limitations. However, at high CO
2
and irradiance, photosynthetic rates were similar between leaf and husk tissue when expressed on a chlorophyll basis.
The CO
2
compensation points of husk tissue were high compared to leaf tissue, but did not increase with increasing O
2
C
3
concentration as is commonly observed in photosynthetic systems suggesting compartmentalisation of the primary carboxylating enzyme Rubisco as it operates in C
4
plants. Both Rubisco activity and the ratio of PEP carboxylase to Rubisco activity were reduced in husk and immunolocalisation measurements indicated Rubisco was present not only in bundle sheath cell chloroplasts but also in mesophyll cells distant from the vascular bundles. However realtime carbon isotope discrimination measured concurrently with gas exchange indicated that the
Rubisco present in distant mesophyll cells of maize husk was unlikely to be contributing significantly to fixation of CO
2
as a low discrimination against 13 C was observed

(~ 4 ‰). From this study we conclude that CO
2 assimilation in maize husk tissue is most likely operating via a C
4 pathway with the expression of Rubisco in distant mesophyll yet to be explained.
C
4
rice – an ideal arena for systems biology research
Zhu, X 1
1 Partner Institute for Computational Biology, Xuhui,
China
Engineering C
4 photosynthetic pathway into C
3
crops has the potential to dramatically increase the yields of major
C
3
crops. The genetic control of features involved in C
4 photosynthesis are still far from being understood; which partially explains why we gained little success in C
4 engineering so far. Next generation sequencing techniques and other high throughput technologies are offering an unprecedented opportunity to elucidate the developmental and evolutionary process of C
4 photosynthesis. Two contrasting hypotheses about evolution of C
4
photosynthesis exist, i.e. master switch hypothesis and incremental gain hypothesis. These two hypotheses demand two different research strategies to proceed in parallel to maximize the success of C
4 engineering. In either case, systems biology research will play pivotal roles in identifying key regulatory controlling development of C
4
features, identifying essential biochemical and anatomical features required to achieve high photosynthetic efficiency, elucidating genetic mechanisms underlining C differentiation and
4 ultimately identifying viable routes to engineer C
As a highly interdisciplinary project, the C
4
4
rice.
rice project will have far-reaching impacts on both the basic and applied research related to agriculture in the 21st century.
Halophytes: what makes them special? Revealing ionic mechanisms of salinity tolerance
Shabala, S 1 , Moreno, AR 2
, Bose, J 1
, Hariadi, Y 3
, Jacobsen, S-E 5
, Mackay, A 1 ,
1
Tian, Y 4
School of Agricultural Science, University of Tasmania,
2 Australia; Unitat de Fisiologia Vegetal, Universitat
Autònoma de Barcelona, Spain;
University of Jember, Indonesia;
3 Dept of Physics,
4 Institute of Grassland
Science, Northeast Normal University, China; 5 Dept of
Agriculture and Ecology, University of Copenhagen,
Denmark
The increasing problem of global land salinisation and associated multibillion dollars losses in crop production require a better understanding of key physiological mechanisms conferring salinity tolerance in crops. The effective way of gaining such knowledge comes from studying halophytes. Halophytes have always attracted the attention of plant physiologists, due to their remarkable ability to tolerate and even benefit from salt concentrations that kill most other plant species. At the very least, halophytes may provide genes that allow transgenic conference of salinity tolerance to crops. In addition, some halophytes have already been tested as vegetable, forage and oilseed crops in agronomic field trials, whilst others show good potential to be developed as crops. Surprisingly, our knowledge of fundamental ionic and molecular mechanisms conferring salinity tolerance in halophytes is rather limited, and at best is restricted to several model species. This talk summarises the current knowledge of physiological mechanisms regulating ion uptake and sequestration in halophytes and provides insights into the identity of membrane-transport systems mediating ion transport in halophyte root and leaf tissues. The focus of this study was on two species: quinoa ( Chenopodium quinoa Willd.) and Atriplex
( Atriplex lentiformis L.). A range of physiological techniques (leaf gas exchange and photosynthetic characteristics; sap osmolality; tissue elemental composition) were used to reveal mechanisms of osmotic adjustment and tissue-specific ion compartmentation in both species at the whole-plant level. It was found that
95% of osmotic adjustment in old leaves and between 80 and 85% of osmotic adjustment in young leaves was achieved by means of accumulation of inorganic ions
(Na + , K + and Cl ) when plants were grown at elevated (up to 500 mM NaCl) salinity levels, whilst the role of organic osmolytes was very limited. Both species also possessed an efficient mechanism to control Na + and K loading into the xylem, as well as for efficient Na
+
+ sequestration in leaves. Whole-plant experiments were complemented by a range of microelectrode studies (noninvasive ion flux measurements; membrane potential; patch clamp) aimed to reveal the identity of specific ion transporters mediating the above process. We compare the kinetics of net K + and Na + fluxes between different root zones (e.g. elongation vs mature zone) and report a differential sensitivity of quinoa and Atriplex root tissues to NaCl and oxidative (hydroxyl-generating Cu/ascorbate mixture) stress. We show that regulation of both depolarization-activated outward-rectifying K + -selective
(GORK) channels and non-selective cation (NSCC) channels are instrumental to halophytes adaptation to saline conditions, and reveal an important role for a H + -
ATPase pump in this regulation. We also demonstrate a feasibility of using the MIFE technique to map ion flux profiles from intact plant leaves and report, for the first time, in situ data on patterns of net K + , Na + and H + flux kinetics from halophyte bladders in response to a range of salinity treatments.
Nitrogen stress-induced alterations in the leaf
1 proteome of wheat varieties
Ahmad, A 1 , Chandna, A 1 , Iqbal, M 1
Molecular Ecology Laboratory, Dept of Botany,
Hamdard University, New Delhi, India
Nitrogen (N) deficiency in agricultural fields is a major limitation that poses a serious threat to the food supply in the world. Differential nitrogen efficiency of 135 wheat genotypes was investigated on the basis of their performance under N-insufficient and N-sufficient conditions in terms of growth and N uptake and assimilation capacities. Changes in the proteome of Nstress sensitive and N-stress tolerant wheat varieties were studied using two-dimensional gel electrophoresis (2-
DE) via liquid chromatography–tandem mass spectrometry (LC-MS/MS) in order to find out the
79

mechanism involved in the differential response of these genotypes to N levels. Forty protein spots were significantly altered by exposure to the nitrogen treatments. Among them, 10 protein spots were downregulated and 30 protein spots were up-regulated. The proteins involved in the energy production/regulation and metabolism in plant leaf tissue were found to be differentially expressed under N treatments. The proteins involved in energy production included those having a role in glycolysis, respiration, electron transport and photosynthesis whereas the proteins involved in metabolism included those concerning the metabolism of sugars, polysaccharides, sterols and cofactors. Our data suggest that Rubisco and glutamine synthetase enzymes might act as suitable targets for genetic manipulation of plants, which can grow and yield well at low nitrogen levels.
Implications of inefficient use of nitrate by sugarcane
Robinson, N 1 , Brackin, R 1
Lakshmanan, P 3
, Vinall, K 1
, Schmidt, S 1
, Rennenberg, H 2 ,
1
2
University of Queensland, St Lucia, Australia;
University of Freiburg, Freiburg, Germany;
Indooroopilly, Australia
3 BSES Ltd,
Recommended N fertilizer rates for sugarcane crops varies greatly between countries, ranging from 120–220 kg N ha -1 in Australia to 750 kg N ha -1 in China. Only
20–40 % of N-fertilizer is recovered in sugarcane systems supplied with 100–200 kg N ha -1 and up to 10% of fertilizer-N is emitted as greenhouse gas N
2
O. To discover reasons for the comparatively low N-fertiliser use efficiency of sugarcane, we examined the use of nitrate and ammonium by sugarcane, its ancestral species and grain crops sorghum and maize. N forms were supplied simultaneously and mimicked the N availability in sugarcane soils. Results indicate that (1) commercial sugarcane varieties and ancestral Saccharum spontaneum under non-limiting N supply incorporated substantially less nitrate (40–45% of ammonium uptake) than related species Erianthus sp., sorghum and maize, and (2) intact roots in a commercial crop incorporated less nitrate than ammonium. We conclude that commercial sugarcane varieties and ancestral species have a much lower ability to use nitrate than related species in the supertribe
Andropogonodea. Our study challenges the paradigm that nitrate is the preferred N source for crops, and provides an explanation for the inefficient N use in sugarcane systems. Potential avenues for improvement of
N-fertilizer use include selection of sugarcane genotypes with better nitrate use and improved fertilizer and soil management aimed to reduce production of nitrate.
Complex organic nutrients as nutrient sources for plants and triggers of root growth
Paungfoo-Lonhienne, C 1
D 2 , Schmidt, S 1
, Lonhienne, TGA 1 , Rentsch,
80
1 School of Biological Sciences, The University of
Queensland, St Lucia, Australia; 2 Institute of Plant
Sciences, University of Bern, Bern, Switzerland
Pollution of land, water and air derived from inefficient use of nitrogen (N) fertilisers is of great global concern.
Phosphorus (P) deposits are rapidly being depleted and accompanying price increases will threaten food security in coming decades. Organic nutrients are increasingly considered as alternatives to currently used inorganic and urea-based fertilisers, but the long-held view remains that plants rely on inorganic forms of nutrients and entirely depend on microbial conversion of soil organic nitrogen and phosphorus prior to uptake into roots. Using green fluorescence protein (GFP) and fluorescent-labelled phosphorothioate oligonucleotides (S-DNA) as sources of organic N and P, we show that protein and DNA are taken up by plant roots. We further show that addition of protein or DNA to nutrient-replete growth medium enhances the growth of lateral roots and root hairs. While uncertainty remains about the mechanisms of uptake, these findings change our view of the spectrum of N and
P sources accessible to plants and also demonstrate that protein and DNA trigger morphological responses. The proliferation of roots in response to the tested organic compounds may enhance access to organic matter-rich patches in the soil. Our research challenges the current paradigm that plants rely on microbes and soil fauna for break-down of organic matter and provide further evidence for heterotrophy in plants.
Role of organic nitrogen in sugarcane cropping
Lakshmanan, P
1
Brackin, R 2
1 , Holst, J
, Schmidt, S 2
2 , Robinson, N 2 , Vinall, K 2 ,
1Bureau of Sugar Experimental Stations, BSES Ltd,
Indooroopilly, Australia; 2 2School of Biological
Sciences, The University of Queensland, St Lucia,
Australia
Sugarcane is a crop of industrial scales and has a great potential of sugarcane to supply food, biomaterials and fuels in future economies. However, sugarcane farming in Australia and elsewhere is accompanied by a large nitrogen footprint with up to 750 kg N applied per hectare per crop cycle. In Australia, up to 70% of the applied N fertiliser in sugarcane cropping is lost to the environment and pollutes waters of the Great Barrier
Reef and causes emissions rates of potent greenhouse gas nitrous oxide which exceed IPCC estimates several fold.
Our research shows that high N losses occur within the first 2-3 months of fertiliser application indicating that plant-N demand and soil-N supply are ill matched.
Analyzing the contribution of inorganic and organic
(amino acids) N forms in the soil over a year's crop cycle, we show that organic N is a likely N source for sugarcane, representing up to 70% of soluble, lowmolecular weight N in soils. We discovered that sugarcane prefers ammonium and amino acids as N sources for growth and discriminates against nitrate when provided with a choice of N sources. In addition to being a potential N source which has a longer resident time in soil than inorganic N, organic N also enhances root branching in sugarcane which could benefit the acquisition of resources from soil. We will present an overview of Australian farming practices including the

techniques of 'trash blanketing' which results in a thick blanket of plant residue on the soil surface after harvest.
How organic N can contribute to sustainable sugarcane farming is discussed.
Improvement in the growth and quality of green onion (
Allium cepa
L.) plants by some bioregulators in the new reclaimed area at Nobaria region, Egypt
Abd El Wahed, M 1 , El-Awadi, M 1
1 National Research Center, Dokki, Egypt
Two field experiments were conducted in the
Experimental Station of the National Research Center at
Nobaria region, Behira Governorate, Egypt during two successive seasons 2007–2008 and 2008–2009. The trial was carried out to study the effect of some bioregulators
(glutathione, cysteine and methionine) on growth, quality and some chemical constituents of green onion ( Allium cepa L. cv Giza 6) plants. Cultivation of onion bulb (1 cm diameter) was performed on 29th Oct. in both seasons. The treatments included one foliar application of glutathione, cysteine or methionine at four concentrations i.e. 0, 25, 50, and 75 ppm after 33 days from sowing.
Results indicated that foliar application of the three bioregulators promoted growth and quality of green onion criteria: shoot length, length of white part, bulb diameter, No. of leaves and fresh and dry weight of plant.
Chemical constituents were significantly influenced by bioregulators treatments especially in plants treated with the lower concentrations (25 ppm) of glutathione, cysteine and methionine. Our results cleared that significantly increment of photosynthetic pigments, fixed oil percentage, total protein, total phenol, and total free amino acids. On the other hand, flavonoid content and total soluble salts were significantly reduced.
Review of developmental and anatomical changes under the influence of cesium uptake in
Chenopodium album
Tajadod, G 1 , Mogoei, R 1 , Mazhar, F 1
1 Islamic Azad University,North Tehran Branch, Iran
Chenopodim album belongs to the Chenopodiaceae family and is an annual plant which can be found in most parts of Iran. This plant has the ability to absorb and refine heavy metals, specially cesium, from the environment. Cesium (Cs) is one of the alkali metals in soil which plants desire to absorb because of its similarity to calcium. While absorption of Cs from the environment, its impacts upon plant developmental and anatomical structure were studied. For this study seeds of
C. album were grown using hydroponic culture with
Hogland solution. After 2 months, seedlings (6 cm height
) were transferred to a medium with different amount of
Cs. Three groups, including one control and two treated groups (watered with 2 and 5 ppm Cs ), were studied.
Considering the ability of absorption of Cs by C. album , and its accumulation in the plant, cells suffer serious injuries in terms of anatomical and developmental structure. By increasing Cs concentration, changes, including destruction and irriguarity of cells, especially in stem meristem layers is headed with shortening of plant height and yellow color of its leaves.
Novel functions of pea MCM6 single subunit as DNA helicase and in promoting salinity stress tolerance without yield penalty
Tuteja, N 1
1 International Centre for Genetic Engineering &
Biotechnology, New Delhi, India
The initiation of DNA replication starts from origins and is controlled by a multiprotein complex, which involves many protein factors. One of the important factors is hetrohexameric minichromosome maintenance (MCM2-
7) protein complex which is evolutionarily conserved and functions as essential replicative helicase for DNA replication. However, to date, helicase activity in the single eukaryotic MCM subunit and the role of MCMs in abiotic stress tolerance in plants have not been reported.
Salinity stress tolerance is an important trait that is required to overcome salinity-induced reduction in plant productivity. Here, we report the following: (a)Pea
MCM6 single subunit (cDNA: 2.98 kb, protein: 93 kDa) alone contains 3’-5’ DNA helicase, ATP-binding and
ATPase activities. (b) The DNA helicase activity was stimulated by replication fork-like substrate. (c) In vitro it forms a homohexamer and is localized in nucleus and cytosol. (d) The MCM6 transcript is upregulated in pea plant in response to high salinity and cold stress and not with ABA, drought, heat stresses. (e) MCM6 overexpression in tobacco plants confers salinity tolerance. The T1 transgenics plants were able to grow to maturity without yield penalty. (f)It was observed that in salt-grown T1 transgenic plants Na + ions is mostly accumulated in mature leaves and not in seeds. These findings provide first direct evidence that single subunit
MCM6 contains DNA helicase activity which is unique to plant MCM6 protein, as the activity was only reported for heteromultimers of MCM proteins in animal system.
These findings also provide first direct evidence that single subunit MCM6 is involved in salinity stress tolerance without yield loss. Overall, this discovery should make an important contribution to a better understanding of DNA replication in plants and the findings also suggest that DNA replication machinery can be exploited for promoting stress tolerance in crop plants.
1
New resources for hybrid rice breeding
Toriyama, K 1
Graduate School of Agricultural Science, Tohoku
University, Japan
Hybrid rice has an average 15% to 20% yield advantage over inbred lines. In China, it is planted on approximately 16 million hectares – more than half of
China’s total rice area of 28 million hectares (Barclay
2007). Other than China, Vietnam, India and Philippines have commercialized hybrid rice. Bangladesh, Indonesia,
Pakistan have also achieved recent success with hybrid rice. Hybrid rice is expected to be a magic wand for world hunger. For hybrid rice breeding, cytoplasmic
81

male sterility (CMS) has been widely utilized. Most of the commercial rice hybrids are based on a single CMS source, the WA (wild abortive) cytoplasm.
Approximately one-tenth of the world rice fields rely on a single CMS-cytoplasm. If WA-CMS-specific disease would emerge, Hybrid rice would be completely destroyed, leading food panic. To avoid the potential threat of genetic vulnerability of rice hybrids, the development of several CMS/Rf systems is desirable. We have been studying three types of CMS in rice: BT-CMS originated from Chinsurah Boro II, LD-CMS from Lead
Rice and CW-CMS from Chinese Wild rice. We have achieved positional cloning of the fertility restorer genes,
RF1 for BT-CMS, RF2 for LD-CMS and RF17 for CW-
CMS. RF1 has been shown to encode pentatricopeptide repeat protein, which binds mitochondrial atp6-orf79
RNA and promote RNA processing, suppressing the translation of cytotoxic orf79 (Kazama et al. 2008). We have shown that RF17 encodes an unknown protein containing a part of acyl-carrier protein synthase-like domain and involved in perception of retrograde signaling from CW-mitochondria and, named RF17 as
RETROGRADE-REGULATED MALE STERILITY
(RMS) (Fujii and Toriyama 2009). We have recently found that RF2 encodes a glycine-rich protein (Itabashi et al. submitted). We have also determined the whole genomic sequences of LD- and CW-mitochondria and found unique ORFs in each genome (Fujii et al 2010).
Elucidation of the RF genes and unique mitochondrial
ORFs will enable us to incorporate LD-CMS/RF2 and
CW-CMS/RF17 systems, as well as BT-CMS/RF1 system, to practical hybrid rice breeding program.
Molecular findings of these mechanisms also provide us useful information for engineering hybrid varieties in various crops for the future.
The reproductive biology of wheat and barley: from developmental genetics to crop improvement
Trevaskis, B 1
1 CSIRO Division of Plant Industry, Canberra, Australia
Advances in molecular biology and functional genomics that have been pioneered in model plant systems are now being applied to the study of reproductive development in cereals. This is providing profound insights into the biological processes that control flowering and grain production in economically important crops such as wheat and barley. Genes controlling the seasonal timing of flowering of temperate cereals have now been isolated, including genes that regulate the vernalization response (the promotion of flowering by prolonged cold during winter) and the long-day flowering-response. This presentation will describe the genes controlling vernalization and daylength flowering-responses in cereals and examine how interactions between these genes regulate the process of floral development in wheat and barley. The historical importance of these genes to the Australian grains industry will be used to highlight how cereal breeders have utilised variation in genes controlling these seasonal flowering-responses to adapt modern varieties to a wide range of growing regions, sowing times and climates. The potential for these genes to contribute to future crop improvement strategies will also be discussed.
82
Study of storage protein composition of barley with a
1 focus on improving the baking quality
Vincze, E 1 , Hansen, M
Langkilde-Lauesen, A 1
1 , Aaslo, P
Aarhus University, Denmark
1 , Falkiewicz, A 1 ,
Barley is grown in many regions throughout the world, in part due to its ability to adapt to less favourable growing conditions. Approximately two-thirds of the worldwide annual barley production is used for feed, one-third for malting and a small (2%) but increasing percentage used as food and in food processing. As barley has the potential to offer considerable human nutritional benefits, there is renewed interest in developing barley for a wider application in the food sector. Considerable focus is on supplementing wheat-based breads with barley to introduce the inherent nutritional advantages of barley flour. Until recently, the most important selection criteria of breeding under modern high input agriculture conditions have been yield. Consequently, the present day cultivars are characterised by unfavourable protein content with inferior leavening and baking properties.
Using knowledge from wheat baking quality parameters, one of the most important storage protein called glutenins has its highly homologue counterpart in barley, called Dhordein. The first step in our study was to gain more knowledge of the storage protein accumulation and AA metabolism in the barley grain. The chosen method was gene expression analysis using cDNA microarrays.
Grain-specific cDNA microarray was used to investigate amino acid biosynthesis and storage protein accumulation in the developing grains of field-grown barley. A distance matrix was constructed which enabled the identification of three clusters corresponding to the early, middle and late phase of grain development. The gene expression pattern associated with the clusters was investigated using a pathway-specific analysis with specific reference to the temporal expression levels of a range of genes involved mainly in the photosynthesis process, amino acid and storage protein metabolism.
Furthermore we have been able to show that different members of the storage protein families, including Dhordeins are synthesized during different stages of grain filling. Studying the importance of D-hordein, the predicted allelic variation and exploiting the substantial genetic variation in storage protein composition in the barley gene pool might enable us improving the baking qualities of barley flour. From our high protein barley collection 49 were selected and grown in field conditions. We analysed storage protein concentration, protein patterns and AA composition and performed small scale baking trails. Our results indicate that there is genetic variation in the relative proportions of the storage proteins affecting the amino acid compositions and baking quality. Some of the cultivars have better baking quality and the better baking quality correlates with the higher MW protein band(s). We isolated these higher
MW protein band(s) from the SDS-PAGE gels and showed by mass spectrometry that the proteins belonged to the D-hordein family. The analysed protein from different cultivars had no major differences on the 5’ or
3’ ends. The highly repetitive middle part of the protein was not accessible by the mass spectrometry studies. The first cloned corresponding genes showed differences in the number of repetitive motifs. These variations could

be further exploited in conventional breeding programs as well as by genetic modification (under progress) to increase this crop baking and nutritional qualities.
Transcriptomics and proteomics to identify component isoenzymes in the metabolic pathway for triricinoleate biosynthesis in Castor bean
Slabas, A 1 , Kroon, J 1 , Brown, A 1 , Simon, W 1
1 University of Durham, Durham, UK
Castor bean [ Ricinus ] oil is a rich source of ricinoleic acid [12-hydroxyoleic acid: 18:1 12-OH] a raw material for the production on N-11, a polymer extensively used by the automotive industry to carry hydraulic fluids at high temperatures. The key gene involved in conversion of oleic acid to ricinoleic acid was identified using an
EST approach in 1995. Expression of the 18:1 12-OH lase in transgenic plants has resulted in up to 20% 18:1
12-OH in seed oil which is well below the 90% level found in Ricinus . To try and identify components involved in the assembly of Triricinoleate in Ricinus we have employed a dual transcriptomic and proteomic approach. Key to the proteomic approach has been the use highly enriched endoplasmic reticulum and the use of
MUDPIT. The availability of the complete genome sequence of Ricinus supplied the necessary framework for identification purposes. Here we will describe our approach to the identification of key isoenzymes which are candidates for the assembly of ricinoleic acid onto the glycerol backbone.
Rice genotypes responses to elevated CO
2 from two free air CO
2
Hasegawa, T 1 , Sakai, H 1 , Tokida, T 1
obtained
enrichment field trials
1
Zhu, C 1 , Usui, Y 1
National Institute for Agro-Environmental Sciences,
Japan; 2 Taiyokeiki Co. Ltd, Japan
Atmospheric CO
2
concentration [CO
2
, Nakamura, H 1 ,
] is an important environmental variable for plant growth, because it is the substrate for photosynthesis. It is less variable than air temperature, solar radiation and precipitation, so the effect of day-to-day or season-to-season variation is not large. However, the increase in [CO
2
] since the industrial revolution has already exceeded 100 ppm (280 ppm in
1800 vs 388 in 2009), and is projected to increase to
470–570 ppm in by the middle of the century (IPCC,
2001). This is the range in which plants are highly responsive. The current level of [CO
CO
2
fixation process of C
3
2
] is limiting to the
plants, so that rising [CO
2
] promotes leaf-level photosynthesis and thereby biomass production and grain yield. This CO
2
fertilization effect is one of few positive impacts on crops by the climate change, which may even change the sign of the overall climate change impacts on crop production. Free-air CO
2 enrichment is so far the best approach in determining the
CO
2
CO
2
fertilization effect under field conditions. However,
responses could be dependent on environmental conditions. Considering a wide range of environmental conditions where rice production is practiced, we need to determine the FACE environment interactions for the results to be widely applicable. Recent rice FACE studies have shown that there is a significant difference in grain yield among genotypes. Identifying the genotypes FACE environment will provide us with invaluable information as to the rice adaption strategy to climate change. We started our rice FACE study at Shizukuishi (39º38' N)
Japan since 1998 but finished in 2008. However, we have just opened a new FACE site at Tsukuba (35º 58' N) in
2010. The two sites differ significantly in the growth temperature conditions; seasonal mean air temperature was 19.9ºC at Shizukuishi and 23.5ºC at Tsukuba. This allows us to test the CO
2
responses of the varieties common to both Shizukuishi and Tsukuba FACE sites. In this study, therefore, we attempted to determine the traits that can confer better CO
2
responsiveness of the rice genotypes based on the two FACE experiments in Japan.
Exacerbation rather than amelioration of drought stress in soybean grown at elevated CO
2
in the field
1
Leakey, A 1 , Gray, S 1
University of Illinois At Urbana–Champaign, USA
By 2050, summer precipitation in the Midwest U.S. is projected to decrease by 5–40%. Concurrently, atmospheric [CO
2
] is projected to increase to at least 550 ppm. Many models of future ecosystem function and food supply assume that elevated atmospheric CO
2
, by reducing stomatal conductance, will decrease plant water uptake and soil moisture depletion, ameliorating the impacts of drought on photosynthesis and productivity.
We tested whether this mechanism operates in fieldgrown soybean exposed to ambient (385 ppm) or elevated (585 ppm) CO
2
combined with either naturally occurring precipitation precipitation (low-H
2
(high-H
2
O) or reduced
O) at the soybean free air CO
2 enrichment (soyFACE) facility in Champaign, IL.
Rainfall was intercepted with retractable awnings to reduce Jun–Sep water inputs into low-H
2
O plots by 57% in 2009 and 32% in 2010. Contrary to the current paradigm, soybean grown at elevated CO
2
responded to soil moisture deficit with greater decreases in stomatal conductance and photosynthesis compared to ambient
CO
2
-grown plants. This was associated with greater root length in shallow, drying soil layers at elevated CO
2
. We are currently testing if greater chemical root-to-shoot signaling was responsible for the enhanced sensitivity of soybean to drought under elevated CO
2
. Greater drought sensitivity can eliminate the yield benefits of elevated
CO
2
, and raises the possibility that current projections of ecosystem function and food supply are overly optimistic.
Will elevated CO
2
change comparative performance of dryland wheat cultivars?
Tausz, M 1 , Posch, S
Bourgault, M
Chapman, S 5
5
2 , Thilakarthne, LC 2
, Dreccer, F 5
, Fitzgerald, G 4 , Seneweera, S 2
, Lakmini, R 3
, Wockner, K 5
,
, Palta, J 5
1 Dept of Forest and Ecosystem Science, Melbourne
School of Land and Environment, The University of
Melbourne, Australia; 2 Dept of Agriculture and Food
,
,
83

Systems, Melbourne School of Land and Environment,
The University of Melbourne, Australia;
Plant Nutrition Institute, Australia; 4 Victorian State Dept of Primary Industries, Australia;
3 International
5 CSIRO Plant
Industries, Climate Adaptation Flagship, Australia;
Wheat ( Triticum aestivum ) cultivars have long been bred to perform under dry and hot conditions, for example, by selecting for traits such as superior transpiration efficiency or low tillering types. With climate change expected to affect rainfall patterns and raise temperatures in dryland agriculture regions, the importance of drought and heat tolerance will most likely increase. On the other hand, elevated atmospheric CO
2
, the ultimate cause for predicted climate change, can have a positive direct effect on crop growth and performance. Cultivars with superior performance under elevated CO
2
will therefore be important for sustainable future crop production, yet there is some evidence that past breeding activities have not resulted in high CO
2 certainty of rising CO
2
responsiveness. Despite the
concentrations, intra-specific variation of crop response to elevated CO
2
has received little attention. We have little information on what particular traits enable maximum gains under elevated
CO
2
, and even less knowledge about how to ensure that optimum responsiveness to elevated CO
2
is compatible with other desirable properties such as superior transpiration efficiency, reliable yield under dry conditions, or stress tolerance. Climate chambers are the ideal set-up to test CO
2
responsiveness of wheat cultivars with specific traits under fully controlled conditions with high throughput capacity. Tunnel exposure systems allow field exposure under semicontrolled conditions including controlled variations in temperature. The Australian Grains Free Air
Carbondioxide Enrichment facility (AGFACE) in the dryland wheat cropping region in Horsham, Victoria, allows exposure of a range of wheat varieties to elevated
CO
2
under realistic field conditions. A combination of results from such different systems will give a robust picture of how different traits govern CO
2
responsiveness of wheat. Contrasting pairs of commercially available wheat cultivars were tested in these systems including: near-isogenic lines Drysdale and Hartog, with Drysdale previously selected for superior transpiration efficiency using carbon isotope ( 13 C) composition. H45 and Yitpi, with H45 selected for reliable performance in dry areas showing lower tillering, and Yitpi a freely tillering type.
In addition to growth and yield parameters, we measured in situ leaf gas exchange and chlorophyll fluorescence to investigate whether elevated CO
2
changes the comparative performance of the contrasting pairs of cultivars in terms of stomatal behaviour, leaf level carbon assimilation, light use and photoprotection characteristics. Such information is important to guide future cultivar selection and breeding programmes.
Acknowledgements: The presented research received funding from The Australian Commonwealth Dept of
Agriculture, Fisheries and Forestry (DAFF) and the
Grains Research and Development Corporation (GRDC).
Effects of free-air CO
2
enrichment on grain quality traits of spring wheat (
Triticum aestivum
)
Hoegy, P 1 , Wieser, H
Schwadorf, K 1
Zhunusbayeva, Dina 1
2 , Selmair, P
, Breuer, J 1
2 , Koehler, P 2
, Franzaring, Jürgen 1
, Fangmeier, Andreas 1
,
,
84
1 Universitaet Hohenhei, Germany; 2 German Research
Centre for Food Chemistry and Hans-Dieter-Belitz-
Institute for Cereal Research, Gemany
Understanding how elevated CO
2
will affect food quality is currently an often overlooked aspect in terms of climate change and food security in the 21st century.
Spring wheat ( Triticum aestivum L. cv. Triso) was thus exposed to elevated and ambient atmospheric carbon dioxide (CO
2
) concentrations (571.0 versus 418.0 µl l-1,
24-h average) using free-air CO enrichment (FACE)
2 technology at Stuttgart-Hohenheim (Germany) in the year 2008 to examine its impacts on crop yield and grain quality. In the high-CO
2
treatment aboveground biomass production and grain yield were promoted by 8.5% and
10.8%, respectively, although these effects were not statistically significant. The thousand grain weight was significantly increased by 5.4% as size distribution was shifted towards larger grains, which may directly relate to a higher market value. In contrast to these positive effects in terms of food quantity, most grain quality traits were adversely affected. Total protein concentration decreased significantly by 7.9% in the high-CO
2 treatment, which is associated with a deterioration of important grain quality parameters for end-use purposes.
Correspondingly, elevated CO
2
resulted in an overall decrease in the concentrations of protein bound amino acids by 4.2 to 7.9%. Amino acid composition remained unaffected as calculated on a per protein basis except for a negative trend for serine. Moreover, CO -induced
2 impacts on the composition of protein fractions are important for bread-making processes due to their association with mixing and rheological properties of dough. Minerals such as calcium and iron decreased significantly and there was a negative trend for magnesium, suggesting that adjustments of the diet may be required to tackle malnutrition if C
3
crops such as wheat provide the primary source of both protein and micronutrients under CO
2
enrichment. The resulting decline in the nutritive value of wheat grains was probably not simply caused by conversion of the extra
CO
2
into carbohydrates such as fructose, raffinose and fructan, the concentrations of which were increased.
Although soluble non-starch polysaccharides constitute less than 3% of the total grain mass, they are important as they substantially contribute to the water binding capacity of flour thus enabling the production of highquality bread. In contrast, starch, as the main compound in grains, was significantly decreased by 5.2% due to elevated CO
2
, which is in accordance with our findings on lower concentrations of total non-structural carbohydrates and carbon. At the same time, concentrations of total lipids, which are essential for the milling properties of flour and also for bread-making quality, remained unaffected by the CO
2
treatment.
Overall, the carbon to nitrogen ratio was significantly increased by 8.5% under CO
2
enrichment. On the basis of the results described before, it is concluded that implications may occur for consumer nutrition and health as well as for industrial processing of wheat grains in a future high-CO
2
world.
Will elevated CO
2
change the form and function of wheat root systems?
Palta, JA
Milroy, SP 2
1 , Benlloch-Gonzalez, M 1 , Bramley, H 2 ,

1 CSIRO, Plant Industry, Wembley, Australia; 2 The
University of Western Australia Institute of Agriculture,
Crawley, Australia
Several studies investigating the effects of elevated atmospheric CO
2
on wheat have concluded that the main effects are increases in photosynthesis, biomass, grain yield and the efficiency of water use. These are effects measured directly in the above-ground parts of the plant.
Measurements of the below-ground parts, particularly the root system, have received little direct attention, despite the root system being a key part of the crop. One of the reasons is because root systems cannot be readily observed in the soil environment. However, the recent use of new experimental techniques to observe root system growth, proliferation, rooting patterns and their functioning will provide opportunities to elucidate the effects of elevated atmospheric CO
2
on wheat root systems. The aim of this paper is to explore the effects that elevated atmospheric CO
2
may have on the form and function of wheat roots systems. In particular we will focus on whether elevated atmospheric CO
2
can affect root system traits for water and nutrient uptake. In answering this question we used published and unpublished data collected from wheat genotypes grown in pots and glass-walled root growth boxes under elevated atmospheric CO
2
in glasshouse conditions. We focus on root systems traits such as increased root distribution at depth, deeper rooting, fast root elongation rates, reduced diameter of the xylem vessel in the seminal roots, root proliferation through branching and improved root:shoot ratio of dry matter.
Acknowledgements: The presented research received funding from CSIRO, Australian Commonwealth
Department of Agriculture, Fisheries and Forestry
(DAFF) and Grains Research and Development
Corporation (GRDC).
Systems model of canopy photosynthesis
Zhu, X 1
1 Partner Institute for Computational Biology, Xuhui,
China
Optimizing canopy photosynthesis, instead of leaf photosynthesis, is critical to gain crop yields. Though leaf photosynthesis and canopy photosynthesis have both been studied extensively in the past, we lack good method to scale from leaf to canopy photosynthesis. This is partly due to the complex canopy architecture and variability of photosynthetic properties inside canopy.
We developed systems models of canopy photosynthesis which include kinetic models of photosynthesis and a three dimensional model of canopy architecture. This model enabled us to study the responses of canopy photosynthesis upon changes in the expression levels of different proteins involved in photosynthesis and related metabolism. This model provides a new framework to enable study the adaptation, optimization and evolution of features associated with canopy photosynthesis under the past, present and future changing climates.
The evolutionary ecology of plant chemical defenses in agroecosystems: past, present and future
McKey, D 1 , Gleadow, R 2 , Cavagnaro, TR 2
1 Centre d'Ecologie Fonctionnelle et Evolutive,
Montpellier, France; 2 School of Biological Sciences,
Monash University, Clayton, Australia
Plant chemical defenses mediate interactions between plants, their herbivores and pathogens, and the natural enemies of these plant parasites. In domesticated plants, farmers are added to this web of interactions. Plant defenses have been subjected to complex selection pressures under domestication, leading both to decreases and to increases in their concentration, and to shifts in their intra-plant distribution. This complexity is explained partly by variation in the kinds of plants farmers chose to domesticate, and partly by the kinds of environments that nature and farmers combined to create in agroecosystems. Today, the interactions mediated by plant defenses are affected by changing climate, rising temperature and increased atmospheric concentrations of carbon dioxide. These changes can threaten food security by lowering not only the production of food but also its nutritional quality. The strategies proposed to counter these threats envisage a wide range of ways, not mutually exclusive, to manage biotic interactions in agroecosystems. These include modifying crop plants’ natural constitutive and induced defenses and other sources of resistance; engineering crop plants to express novel defenses; and increasing the effectiveness of
‘integrated pest management’ (through natural enemies, other components of agrobiodiversity and pesticides) by modifying the structure and functioning of agroecosystems. In this presentation, we examine these proposed solutions through the evolutionary ecologist’s lens: What are their respective costs and benefits, and how do these affect the ecological situations in which each could help farmers produce sufficient amounts of high-quality food? How can the coevolutionary dynamics of interactions between crop plants and their biotic environment be managed to avert catastrophic fluctuation of yield?
Growing populations under changing climates call for more nutritious crops
Johnson, AAT 1,2 , Kyriacou, B 1 , Stangoulis, J 3 , Tester,
1
M 2
School of Botany, The University of Melbourne,
Australia; 2 Australian Centre for Plant Functional
Genomics, Glen Osmond, Australia; 3 Flinders University of South Australia, Australia
The ‘Green Revolution’ that occurred between the 1940s and 1970s saw an enormous increase in agricultural productivity worldwide. The adoption of semi-dwarf and hybrid cereal varieties, as well as improved irrigation, fertilizer and pesticide practices, greatly increased food production in developing countries of the world.
However, not all changes brought about by the Green
85

Revolution were desirable. Cereal crops such as rice and wheat, while high in calories, do not contain significant quantities of nutritionally important micronutrients such as iron (Fe), zinc (Zn) and Vitamin A. As increasing numbers of people have come to rely on cereal crops to meet their energy needs we have seen a reduction in the number of people suffering from caloric hunger but a massive increase, in fact billions of people, suffering from micronutrient hunger – a nutritional disorder often referred to as ‘hidden hunger’ because the effects, while devastating, are not always apparent at first glance. The effects of climate change are likely to further compound this problem. In particular, rising atmospheric carbon dioxide concentrations are predicted to decrease the protein concentration of most crops. Additionally, the concentrations of many essential elements in food crops, such as Fe and Zn, are predicted to decrease as a result of increased atmospheric carbon dioxide. These findings suggest that climate change may further compound severe micronutrient malnutrition problems already affecting billions of people throughout the world and highlight a need for the development of more nutritious crops. Because rice ( Oryza sativa L.) serves as the primary source of calories for more than half of the world’s population, we are working to improve the micronutrient content of the white rice grain.
Nicotianamine (NA) is a natural metal cation chelator that plays important roles in metal acquisition and transport in all plants. To generate rice lines with increased levels of Nicotianamine, the three rice
Nicotianamine synthase (NAS) genes – OsNAS1,
OsNAS2, and OsNAS3 – were constitutively overexpressed using the 35S cauliflower mosaic promoter. Elemental analysis showed that the resulting rice lines contain three-fold more Fe and two-fold more
Zn in white rice. Seed Nicotianamine content is tightly correlated with Fe and Zn content, providing strong evidence that increased Nicotianamine levels are responsible for the observed increased loading of Fe and
Zn into white rice grain. The levels of Fe in these biofortified rice varieties could meet the recommended daily allowance of Fe in a daily serving of rice. We anticipate that biofortification of all major food staples will take on an increasingly vital role as the human population continues to grow under changing climatic conditions.
Abiotic factors alter defense of cyanogenic lima bean
(
Phaseolus lunatus
L.) against a natural insect herbivore
Ballhorn, DJ 1 , Hegeman, AD 1 , Schmitt, I 2 , Kautz, S 1
1 University of Minnesota, Dept of Plant Biology, St Paul,
2 USA; University of Minnesota, Dept of Horticultural
Science St Paul, USA; 3 Biodiversity and Climate
Research Centre BiK-F, Frankfurt, Germany; 4 Goethe
University Frankfurt am Main, Frankfurt, Germany;
5 Field Museum of Natural History, Dept of Zoology,
Chicago, USA
Plants cannot escape when being attacked or exposed to unfavorable conditions, but they are far away from being helpless organisms. In the last decades, research has demonstrated how plants can actively up-regulate a wide array of defensive traits when subjected to stress factors.
However, effects of changing environmental conditions
86 on variability of plant defensive traits and consequences of this variability for higher trophic levels are still little understood. We used wild lima bean (Fabaceae:
Phaseolus lunatus L.) to analyze effects of various abiotic factors (drought and salt stress, nitrogen availability, and CO
2
) on the expression of two exemplary chosen plant defensive traits: cyanogenesis
(nitrogen-based) and phenolics (carbon-based). In growth chamber experiments, exposing cyanogenic lima bean plants to drought stress, soil salinity as well as enhanced nitrogen-availability resulted in a quantitative upregulation of cyanogenic precursor concentration in leaves by approximately 12, 7, and 9%. In contrast to plant cyanogenesis, concentration of phenolics remained unaffected by the treatments. Feeding trials with Mexican bean beetles (Coccinellidae: Epilachna varivestis Muls.), a specialist herbivore, showed that the up-regulation of plant cyanogenesis resulted in a significantly improved defense against larvae and adult beetles. While the effects of drought, salt stress and nitrogen availability on cyanogenesis were observed among leaves of different age, treating plants with enhanced CO
2
(ambient conditions to serve as control as well as 500, 700 and
1000 ppm CO
2
) atmospheres resulted in a distinct chemical responses depending on leaf developmental stage. While under elevated CO
2
the amount of cyanogenic precursors decreased in young, intermediate, and mature leaves, the concentration of total phenolics was significantly increased in intermediate and mature leaves. As a result of this dramatic change in chemical composition among different leaf developmental stages, in feeding trials with Mexican bean beetles we observed a complete shift from highest herbivore damage in mature leaves in controls to highest damage of young leaves under elevated CO
2
. Using cyanogenic lima bean as model plant we could demonstrate that (i) different abiotic parameters quantitatively affect the plant chemical phenotype, that (ii) changes of plant defensive traits can have strong impacts on natural insect herbivores, and (iii) that changes in plant chemical phenotype and effects on herbivores can depend on ontogenetic development of specific plant organs. Since in the long run preferred consumption of young leaves as we observed under elevated CO
2
concentrations can strongly affect plant fitness, effects of abiotic factors on different leaf stages should receive highlighted attention in future research.
Nutrient supply, below ground processes and elevated
CO
2
change the partitioning of resources between growth and defence in cyanogenic plants
1
Gleadow, R 1
S 2 , Miller, R 1
, Cavagnaro, T 1 , Isherwood, S
Monash University, Australia; 2
1 , Seneweera,
The University of
Melbourne, Australia
The challenge for agriculture in the 21st century is to increase productivity in the face of rising global concentrations of atmospheric CO
2
, higher temperatures, reduced water availability, and possibly in conjunction with more expensive fertilisers. Our focus is on the changing balance between plant growth and resource allocation with climate change and its impact on food security. Plants grown at elevated CO
2
experimentally have lower leaf nitrogen and typically allocate less of

that N to protein and photosynthesis. There is very often a concomitant increase in concentration in secondary metabolites. This is expected to decrease the nutritional quality of pastures. We are developing Trifolium repens
L. (clover) as a model species suitable for high throughput experiments in temperate Australia as a companion to our work on the tropical crops, sorghum and cassava. Clover is one of over 2000 species
(including many crops) that produces cyanogenic glycosides, which release hydrogen cyanide (HCN) in response to plant tissue damage (cyanogenesis), primarily as a defence against hervicores. Clover is polymorphic for cyanogenic glycosides and represents an ideal system to strengthen knowledge of defence chemistry, allocation and resource trade-offs under different environmental conditions. It forms symbiotic associations with mycorrhizae as well as Rhizopus , both of which enhance nutrient uptake. Previous contained environment experiments showed that clover allocates more N to endogenous cyanogenic glycosides when grown at elevated CO
2
but only when phosphate is added. We grew clover (cv Haifa) plants for 5 months at ambient and elevated CO
2
under field conditions in the
AGFACE facility at Horsham. Half the plants received supplemental water. In a parallel experiment we grew cyanogenic and non-cyanogenic clover in agricultural soil in a glasshouse and supplied them with fertiliser containing different proportions of N and P. Plant growth and colonisation rates were determined and leaves analysed for cyanogenic glycosides, nitrogen, phosphorus and micronutrients. Ratios of stable isotopes of N were used to indicate the proportion of N in the plant taken up through N fixation, and C isotopes were used to measure the degree of water stress. We found that the change in relative proportions of Carbon, Nitrogen and Phosphorus affect the allocation of resources to defence. We were, however, unable to detect any cost to the plants (in terms of growth sacrifice) in the production of cyanogenic glycosides. Understanding what drives changes to resource allocation in plants, and particularly the role of below ground processors, will be crucial in determining successful management of agriculture and animal health into the future.
FTIR spectroscopy for characterisation of sorghum cell wall variants
Martin, A 1
Grof, CPL 1
, Byrt, CS 1 , Furbank, RT 2 , Sirault, XRR 2 ,
1 University of Newcastle, Australia; 2 CSIRO Plant
Industry and High Resolution Plant Phenomics Centre,
Australia
Sorghum bicolor cell wall composition was investigated using Fourier Transform Infrared (FTIR) spectroscopy and Focal Plane Array (FPA)-FTIR. Cell walls from stem tissue of a number of Sorghum cell wall variants, including reduced lignin (Brown midrib, Bmr) lines, were isolated and FTIR spectra were collected. The digestibility of the isolated cell wall samples were determined by a cellulase digestion method and the composition of isolated cell walls, such as Acetyl bromide lignin, cellulose and hemicellulose content were measured. FTIR absorbance peaks were assigned in accordance with the literature. Peaks associated with lignin and cellulose differed for the Bmr lines as compared with the wild type lines, consistent with wet chemistry data. Pre-treated spectral data was analysed by principle component analysis (PCA). Examination of the loadings plot for the principal components revealed which peaks most influenced the PC scores. FTIR data clearly distinguished differences in cellulose, lignin, and lignin monomers in the Sorghum variants. The correlation between peak assignment data, collected by
FTIR spectroscopy and data for cell wall components such as cellulose, total lignin, lignin monomers and suberin, measured using traditional methods, was investigated. Spectral differences were mapped in stem sections, using FPA-FTIR, and the images were compared with data from histological staining, including berberine, calcofluor white and mäule staining. A partial least squares (PLS) predictive model was developed which incorporated spectral and cell wall digestibility data. For an independent validation set of samples the model accurately predicted the digestibility of the
Sorghum stem tissue from spectral data. This model may be useful for high-through-put screening of Sorghum germplasm to identify variation in cell wall recalcitrance to digestibility.
A free air CO
2
enrichment (FACE) study of the important staple food cassava (
Manihot esculenta
)
Miller, RE
RM 1
1,2 , Rosenthal, DM 3 , Slattery, B 4
, Cavagnaro, TR 1,2 ,Fauquet, CM 6 , Ort, DR
, Gleadow,
3,4,5
1 School of Biological Sciences, Monash University,
Australia; 2 Australian Centre for Biodiversity, Monash
University, Australia; 3 Global Change and
Photosynthesis Research Unit, Agricultural Research
Service, United States Dept of Agriculture, Urbana,
USA; 4 Depts of Plant Biology and 5 Crop Sciences,
University of Illinois, Urbana, USA; 6 International
Laboratory for Tropical Agricultural Biotechnology,
Donald Danforth Plant Science Center, St Louis, USA
Cassava ( Manihot esculenta Crantz., tapioca, manioc) is an important staple food for over 800 million people in
Africa, South America, Asia and the Pacific. Tubers are the main agricultural product, although leaves are also eaten. Globally, up to 70% of cassava is used for food, but cassava is particularly important in sub-Saharan
Africa. Sometimes referred to as the drought, war and famine crop of the developing world, cassava yields well under low nutrients and dry conditions, and is easy to propagate. Consequently, its production worldwide has doubled in the past 30 years, reaching 250 million tonnes in 2009, due largely to increases on small-scale subsistence farms in Africa, but also to greater cultivation for industrial applications. While cassava is an incredibly important source of carbohydrates, it is low in nutrients and protein, and also contains cyanogenic glycosides that release the respiratory toxin hydrogen cyanide. Many cultivars require processing to reduce cyanogen content before consumption, but intake of insufficiently processed cassava can result in chronic medical conditions such as permanent paralysis of the legs (Konzo). Given the importance of cassava to food security, it is essential that we understand how this crop will respond to global climate change, both in terms of yield and chemical composition. The few studies of the
87

response of cassava to increasing atmospheric CO
2
(eCO
2
) to date – conducted in glasshouses or small chambers – have produced contrasting findings. While one study reported an increase in tuber yield, a recent study reported a decrease in tuber yield under 550 and
710 ppm CO
2
. Furthermore, in the latter study, increases in the toxicity of tissues were also reported under eCO
2 suggesting the prospect of both reduced yield and nutritive value under future CO
2
concentrations. It is now well established that the extent of stimulation of C yields by eCO
2
3
crop
varies not only with species and environmental conditions, but also with experimental methodology. Clearly, there is a need for experimentation to assess how cassava might respond to elevated CO
2
under fully open air CO
2
enrichment
(FACE). We conducted a field-based eCO
2
study of cassava at the soyFACE facility, Illinois, USA, as there are currently no FACE sites in areas where cassava is cultivated. Physiological, growth and biochemical responses of cassava to eCO
2
(585 ppm) were measured.
In the fertile soils at soyFACE, cassava responded as typical C
3 to eCO
2
plant, showing increased biomass in response
. In addition, there were significant changes in the nutritional quality of both foliage and tubers. The consequences of the growth and biochemical responses of cassava to eCO
2 food security.
will be discussed in relation to future
Plant signalling networks underpinning crop yield under environmental stress
Davies, W 1
1 Lancaster University, UK
Soil drying commonly limits plant productivity even when there is still a substantial amount of water in the soil. This limitation occurs, initially at least, through an impact on both gas exchange and canopy development and if we are to minimise these restrictions of growth and yielding, it is important to understand their mechanistic basis. We highlight the importance of the influence of both hydraulic and chemical limitations of functioning, growth and development of plants in drying soil and suggest that by processing and responding to these signals, plants are able to optimize growth and development relative to the amount of water and nutrient available in the soil. We propose that via enhancement or suppression of different ‘root signal’ cascades we can intervene to sustain plant yielding under different environmental stresses (Morison et al. 2007). We review the identity of signals limiting leaf conductance, leaf growth and harvest index (e.g Foulkes et al. 2010) and consider prospects for manipulation of signalling, with special reference to the impact of changes in sap pH, modified fluxes of abscisic acid (ABA) and of the ethylene precursor ACC (e.g. Jia and Davies 2007;
Wilkinson and Davies 2009). We can use both synthesis and sensitivity mutants and transgenics to investigate the genetic manipulation of signalling in agriculture but there are many other ways to modify stress signalling cascades
88 for the benefit of crop yield. For example, management techniques such as deficit irrigation, fertilizer applications and buffer treatments can all impact long distance chemical signalling. Such techniques can be operated in conjunction with plant improvement to sustain yielding and increase water use efficiency in dryland agriculture.
Root thermotolerance mechanisms from whole-root
1 to molecular biology
Huang, B 1
Rutgers University, New Brunswick, USA
Heat stress is becoming an increasingly significant limitation to the growth of temperate plants due to global warming. Roots are more sensitive to heat stress than shoot growth due to their lower temperature requirements for optimal growth. Understanding mechanisms of root thermotolerance is of great significance for developing heat-tolerant plants that can adapt to areas with elevated temperatures. This presentation will review current research in mechanisms of root-mediated heat tolerance from whole-plant to molecular biology. From the wholeplant level, root dieback precedes growth decline and leaf senescence under heat stress, suggesting that root systems play critical roles in sensing high soil temperatures and regulating whole-plant responses to heat stress. Root responses to heat stress are reflected in changes of water uptake, nutrient uptake, and hormone synthesis. Along the three major root functions, hormone metabolism is most sensitive to heat stress. Roots are primary sites for the synthesis of some plant hormones, such as cytokinins. Cytokinins metabolism in roots may affect leaf responses to heat stress by regulating leaf senescence and other physiological processes, including leaf photosynthesis. Roots depends on shoots for carbon supply, therefore, efficient carbon utilization could play important role for maintaining root survival under elevated temperatures. Heat tolerance in root systems is also associated with changes at proteomic, metabolic, and molecular levels. The specific proteins, metabolites, and genes associated with the whole-plant physiological changes and superior heat tolerance will be discussed.
Altering root architecture of wheat to increase the
3
2
1 water productivity in India and Australia
Wasson, AP
Prasad, S
CSIRO
4
1 , Richards, RA
, Singh, SK
Christopher, J 5
2 , Rebetzke, GJ
, Watt, M 1
Plant Industry,
1 , Chatrath, R
1
2 , Misra, S
, Kirkegaard, JA 1
3 ,
,
Canberra, Australia;
Directorate of Wheat Research, Karnal, India;
Agharkar Research Institute, Pune, India; 4 Indian
Agricultural Research Institute, Regional Wheat
Research Station, Indore, India; 5 Dept of Employment
Economic Development and Innovation, Leslie Research
Centre, Toowoomba, Australia
India and Australia produce approximately 100 million tonnes of wheat per year. Almost all of the 75 Mt grown in India is consumed within country; over half of the approx 25 Mt grown in Australia is exported such that wheat is Australia’s most important soft commodity.
Both countries are focused on increasing crop yields per

unit available water to meet future food demands. The aim of our joint research is to identify and validate new wheat lines that have faster growing, deeper root systems that capture more water around flowering and grain development in both Australia and India. Modelling and field evidence has shown that relatively small increases in root system descent rate and root length density at depth can lead to up take of an extra 10 mm of water that is converted into 0.5 tonnes of grain – a very high water productivity. The approach is largely field-based because mature wheat roots cannot be selected in the laboratory, and the mechanisms and genes regulating deep root growth are not known. Diverse wheats being evaluated include unique CSIRO lines with double the shoot and root vigour of released varieties in controlled conditions, lines with modified shoot and root architecture through tiller inhibition, prolonged green-leaf duration, synthetic wheats, and varieties from India and Australia with historically high yields in water-limited regions. A novel planting configuration is used with one line per square meter for high density and replication and uniform water availability per line. Non-destructive imaging methods are used to measure leaf transpiration and green leaf area, and root water uptake is measured using electrical resistivity tomography. At the end of the season roots are measured directly by coring to 2 metres in Australia and
India then relate to shoot measurements. An objective is to develop and validate a rapid non-invasive method for breeders. The best lines for further study are narrowed down annually using a ranking algorithm. These lines will be used to identify the mechanisms underlying root architecture variation and to quantify the water productivity gain in India and Australia. In future they may be used for the development of mapping populations to identify molecular markers. Two seasons at six sites in
India and Australia indicate more than two fold variation in the root and shoot parameters.
Hormonal changes in rice plants under alternate wetting and drying irrigation
Yang, J 1 , Zhang, H 1 , Huang, Z 1 , Zhang, J 2
1 Key Laboratory of Crop Genetics and Physiology of
Jiangsu Province, Yangzhou University, Yangzhou,
Jiangsu, China; 2 Dept of Biology, Hong Kong Baptist
University, Hong Kong, China
Plant hormones may reflect soil water status and mediate rice ( Oryza sativa L.) growth and development. This study investigated the changes in hormonal levels in rice plants and their relations with tillering, panicle initiation, and grain filling under alternate wetting and drying irrigation. Two rice cultivars were field grown. Three irrigation regimes, alternate wetting and moderate soil drying (WMD), alternate wetting and severe soil drying
(WSD), and conventional irrigation (CI, continuously flooded), were imposed during the whole growing season. The WMD significantly increased, whereas the
WSD significantly decreased, productive tiller number, spikelet number per panicle, grain filling rate, and grain yield when compared with the CI. Abscisic acid (ABA) levels in root bleedings, leaves, and spikelets were significantly increased during the soil drying period under both WMD and WSD regimes. During the soil drying period, levels of ethylene and 1aminocylopropane -1-carboxylic acid (ACC) were decreased under the WMD, but they were increased under the WSD. The WMD increased, while the WSD reduced, the ratio of ABA to ACC (ABA/ACC). Levels of zeatin (Z) + zeatin riboside (ZR) in both root bleedings and shoots (spikelets and leaves) at the soil drying time under the WMD were comparable with those under the CI, but they were significantly increased when plants were re-watered. The WSD significantly reduced
Z + ZR contents in root bleedings and shoots either during the soil drying or during the re-watering period.
Level of indole-3-acetic acid (IAA) and gibberellins
(GA1 + GA4) in rice plants were significantly reduced under the WSD, and they showed no significant difference between WMD and the CI regimes. Productive tiller number, spikelet number per panicle, grain filling rate, and grain weight were very significantly correlated with the ratio of ABA to ACC and Z + ZR levels in plants. The results suggest that an increase in ABA/ACC in plants during the soil drying period and an enhancement in cytokinin levels during the re-watering time contribute to increases in rice growth and grain yield under a moderate wetting and drying regime.
Exploiting rhizobacterial impacts on hormonal rootto-shoot signalling to ameliorate water stress
Dodd, I 1 , Arkhipova, T 2 , Belimov, A 3
1 Lancaster University, UK; 2 Ufa Science Centre, Russia;
3 All Russia Research Institute of Agricultural
Microbiology, Russia
Soil drying alters the synthesis, metabolism and xylem loading of various plant hormones (1). Plant growth promoting rhizobacteria (PGPR) are commonly found in the rhizosphere (adjacent to the root surface) and may promote plant growth via several diverse mechanisms, including the production or degradation of the major groups of plant hormones that regulate plant growth and development. Although rhizobacterial production of plant hormones seems relatively widespread (as judged from hormone measurements of bacterial culture media), there is increasing evidence (particularly from seedlings grown under gnotobiotic conditions) that rhizobacteria can modify plant hormone status (2). This not only has local effects on root elongation and architecture, thus mediating water and nutrient capture, but can affect hormonal root-to-shoot signalling that alters shoot hormone status and regulates leaf growth and gas exchange. Water stress can stimulate production of the plant hormone ethylene, which is often inhibitory to root and shoot growth. Soil inoculation of pea ( Pisum sativum ) seedlings with the PGPR Variovorax paradoxus
5C-2 containing the enzyme ACC deaminase (which breaks down the ethylene precursor ACC into carbon and nitrogen sources for bacterial growth) attenuated an increase in xylem ACC concentration caused by soil drying, thereby promoting vegetative growth, and increasing plant yield (3). The plant hormones known as cytokinins stimulate cell division and elongation and retard leaf senescence, yet water stress decreases plant cytokinin concentrations. Soil inoculation of lettuce
( Lactuca sativa ) seedlings with the cytokinin-producing
PGPR Bacillus subtilis IB-22 doubled shoot cytokinin concentrations presumably by enhancing root-to-shoot cytokinin transport, and promoted shoot vegetative growth (and thus plant yield) of plants grown in drying
89

soil (4). Further work is required to substantiate whether individual PGPR affect root-to-shoot signalling of more than one hormone class, and whether these systemic effects described under controlled environment conditions occur when field crops are inoculated.
Nevertheless, rhizobacterial amendments offer the potential to ameliorate deleterious effects of water stress on shoot hormone status and thence crop yield (5).
Acknowledgements: We thank BBSRC, DEFRA, RFBR and the Royal Society for supporting work in our laboratories. (1) IC Dodd (2005). (2) IC Dodd et al.
(2010). (3) AA Belimov et al. (2009). (4) TN Arkhipova et al. (2007). (5) IC Dodd (2009).
Effects of soil organisms on aboveground multitrophic interactions are consistent among plant
1 genotypes mediating the interaction
Kabouw, P 1 , Kos, M 2 der Putten, W 1
, Vockenhuber, E
, van Dam, N 1
3 , Biere, A 1 , van
Netherlands Institute of Ecology (NIOO-KNAW), Dept of Terrestrial Ecology, The Netherlands; 2 Wageningen
3
University, Laboratory of Entomology, The Netherlands;
Georg-August-University, Agrarökologie, Germany
In an experimental study we demonstrate that manipulation of interactions between plants and soil organisms can profoundly affect the herbivore load aboveground. To assess whether the outcome of these below–aboveground interactions can be affected by plant genotype we selected two white cabbage ( Brassica oleracea var. capitata ) cultivars. These cultivars were known to differ in their resistance against below- and aboveground pests due to their different profiles of secondary metabolites, including glucosinolates.
Belowground we inoculated soils of the cultivars either with nematodes or micro-organisms and included a sterilized soil as a control treatment. Aboveground we monitored population development of aphids and fitness parameters of parasitoids as a bio-control mechanism.
Soil treatments affected aphid population sizes; microorganisms increased and nematodes tended to decrease aphid population growth. Soil treatments did not affect parasitoid performance, thus not decreasing their efficiency for bio-control. The glucosinolate gluconasturtiin was lacking from one of the cultivars, making this cultivar more susceptible to plant feeding nematodes. Nevertheless, the relative effect of soil treatment on aphid population growth or parasitoid performance was similar for both cultivars. This indicates that, although the soil community did affect the aboveground community, this effect was independent of the cultivar that was mediating the interaction.
Additionally the effect of cultivar on the aboveground community was overall larger than soil treatment effects.
The two cultivars sustained different aphid population growth rates and the parasitoids had a higher mass and a faster developmental time on the cultivar that had the highest aphid population growth. In conclusion, in our study altering belowground interactions consistently altered herbivore loads aboveground.
Inflorescence development in maize
1
McSteen, P 1
Division of Biological Sciences, University of Missouri-
Columbia, Columbia, USA
Maize ( Zea mays or corn) produces two types of inflorescence – the male inflorescence or tassel, which produces the pollen and the female inflorescence or ear, which produces the kernels and is harvested. An understanding of yield potential requires an understanding of the development of the inflorescence and in particular the activity of meristems within the inflorescence. We are using a genetic approach to determine the mechanisms regulating inflorescence development in maize. The spikelet (a small branch bearing the florets) is the fundamental unit of yield in all cereals. Maize and all species in the tribe Andropogoneae produce paired spikelets while grasses outside this clade produce single spikelets. To identify genes controlling spikelet number in grasses, we have been identifying maize mutants which produce single spikelets. One class of these mutants have defects in the biosynthesis, transport or response to the plant growth hormone, auxin, implicating auxin in the process of axillary meristem initiation and spikelet production. This class of mutant often produces no ear shoot, implicating auxin also in the initiation of the ear in maize. Another class of mutants have smaller meristem size implicating the signaling pathways regulating meristem size in the control of spikelet number in the ear. A third class of mutants have defects in nutrient metabolism implicating essential nutrients in the maintenance of the apical meristem and hence ear size. An understanding of the process of inflorescence development is essential for efforts to sustain and improve yield in all cereals. Our current understanding of the mechanism of inflorescence development in maize will be presented.
Specification of spikelet meristem identity directly influences the inflorescence form of rice
Kyozuka, J 1
1 Graduate School of Agriculture and Life Sciences,
University of Tokyo, Japan
Number of grains, one of major determinants of yield in grass species, depends on the inflorescence form.
Flowers of grass species are formed in spikelets and the inflorescence form is described as the pattern of spikelet formation. During rice inflorescence development, panicle branch formation precedes spikelet formation. A delay of spikelet identity specification leads to prolonged branch formation, which results in the formation of bigger inflorescences. On the other hand, precocious specification of the spikelet identity causes production of small inflorescences containing less spikelets. We identified TAWAWA 1(TAW1) and PANICLE
PHYTOMER 2 (PAP2) genes as novel key players in control of the spikelet meristem identity in rice. Semidominant taw1-D1 and taw1-D2 mutants exhibit a highly
90

branched inflorescence phenotype, indicative of the delayed specification of the spikelet meristem identity.
TAW1, encoding a nuclear protein of unknown function, is expressed in the SAM during vegetative phase and its expression level gradually decreases after transition to the reproductive phase. Higher levels of TAW1 mRNA accumulate in taw1-D1 and -D2 mutants and the severity of the phenotype well correlates with the level of TAW1 expression. Based on these observations, we propose
TAW1 as a negative regulator of the spikelet meristem fate in rice. PAP2 is a positive regulator of spikelet meristem identity. Loss of PAP2 function causes an increase of the inflorescence branch formation indicative of the delayed specification of spikelet identity. PAP2 encodes OsMADS34, a member of SEP subfamily
MADS box proteins. Phylogenetic analysis indicated that
SEP subfamily is expanded in grass species and PAP2 is in a grass specific sub-clade of the SEP genes. Our analysis suggested that PAP2 is central in the control of early stages of rice inflorescence development. The genetic interaction between TAW1, PAP2 and other genes which are involved in the control of spikelet identity will be discussed. Possible application of TAW1 and PAP2 genes for rice breeding to increase yield will be also discussed.
Molecular pathways controlling the response to high temperature in cereals
Hemming, MN 1 , Dennis, ES 1 , Trevaskis, B 1
1 CSIRO Plant Industry, Canberra, Australia
Rising global temperatures have the potential to decrease yields in staple crops such as cereals. High temperature accelerates flowering in cereals, but nothing is known about the molecular pathways controlling this response.
Using barley as a model, we are examining the effect of increasing ambient temperature on vegetative and reproductive development in cereals at the physiological and molecular level. Plants are grown under highly controlled and replicable growth conditions in the High
Resolution Plant Phenomics Centre in Canberra to screen germplasm for variability in response to temperature and to conduct gene expression analysis. We aim to identify genes controlling the developmental response to high temperature. I will present a model for the way in which changes in gene expression control reproductive development in response to increasing ambient temperature and describe the contribution of key flowering-time genes to development under high temperature. Finally, I will discuss how identification of key genes controlling the response to increasing temperature in cereals will enable selection of allelic variants that provide positive adaptation to specific climatic conditions.
Genetic regulation of spike development and yield potential in wheat
Swain, S 1 , Cavanagh, C 1
1 CSIRO, Canberra, Australia
Cereal yield potential is determined by a combination of the number of grains formed in a given area and the average grain weight. The number of grains produced by each plant is determined by the number of fertile florets that develop, each of which has the potential to form a single grain. Consequently, a major limit on yield is the number of fertile florets/grains that develop. Fertile florets per plant is a major target for genetic improvement of cereals, including wheat. In wheat, florets are produced by groups of approximately 20 specialized branches (spikelets) which together form a structure known as a spike. The number of florets produced by each spikelet, and the number of spikelets per spike, are under strong genetic (and possibly epigenetic) control. Altering spike architecture, by modifying both the number and position of spikelets, and the number of fertile florets produced by each spikelet, can potentially increase the number of florets/grains per spike, and hence yield potential. We are using a unique wheat mapping population that segregates for variation in several aspects of spike development, including both spikelet number and florets per spikelet, to investigate the relationship between spike architecture and yield potential. One trait we are investigating is the formation of supernumerary spikelets which can form adjacent to normal spikelets leading to a pair of spikelets at a single rachis node of the spike. Detailed analysis reveals that the formation of these paired spikelets varies along the length of the spike, and their number is also affected by different growing conditions, suggesting a link between this trait and assimilate partitioning to the developing spike. Quantitative Trait Locus (QTL) analysis reveals several major loci that control paired spikelet formation and ongoing work is aimed at identifying the genes involved based on the function of rice and maize genes that regulate related processes in these crops.
1
High temperature stress and plant reproduction
Sage, T 1
Dept of Ecology & Evolutionary Biology, University of
Toronto, Canada
One consequence of global climate change is the detrimental effect of increased frequency of high temperature stress (HTS) on crop yields. The primary focus of this research is to assess the impact of HTS
(32ºC and 36ºC) on morphological yield components contributing to Harvest Index in the eudicot, Arabidopsis
(accessions Ler and Cvi) and the monocotyledonous species, Brachypodium distachyon and Oryza sativa .
HTS has a negative impact on Harvest Index all species by significantly reducing initiation of vegetative branching and flowers. In Arabidopsis , 32ºC reduces seed set/silique in Cvi to almost zero whereas Ler still produces a high number of seed/silique. Pollen deposition is directly correlated with reductions in seed yield/silique such that 32ºC almost eliminates pollen deposition in Cvi but not in Ler. Pollen deposition, and hence seed production, is also severely reduced at 32ºC in Oryza and Brachypodium . HTS of 36 C eliminates seed set for all species examined. In comparison to processes leading to reductions in seed set at 32ºC, an absence of seed set at 36ºC results from termination of microspore mother cell development prior to meiosis or abortion of uninucleate microspores. Developmental studies indicate that miscarriage during maturation of microspore mother cells and uninucleate microspores involves autophagic cell death. Analysis of reproduction
91

at 32ºC in LerXCvi Recombinant Inbred Lines has identified QTL involved in filament and anther development that play an important role in anther dehiscence and pollen deposition. Within these QTL are genes involved in cell division and elongation, ethylene response and signaling, and heat shock. With the use of cryofixation to examine ultrastructural features of anther development at 32ºC in Oryza and Brachypodium , it has been determined that HTS prevents anther indehiscence by interfering with endothecial contraction and not the development of the stomium and pollen swelling. These results shed light on the stages of reproduction that are most sensitive to HTS in both a eudicot and monocots;
HTS has an unfavorable impact on Harvest Index by interfering with male reproductive development and suppressing vegetative and reproductive meristem development. QTL that influence these developmental processes in Arabidopsis will be of interest for future studies aimed at improving crop yield of both eudicots and monocots under increased global temperatures.
Bioprospecting flavonoid and terpenoid pathways in target plants tea,
Picrorhiza and
Arnebia
Singh, K 1 , Rani, A 1
M 1 , Dutt, S 1
Sharma, M 1
, Kawoosa, T
, Kumar, S 1
, Ahuja, PS 1 ,
1 ,
Kumar, S
Singh, RS
, Gulati, A 1
1
1 , Mahajan,
, Bhushan, S 1 ,
1 Biotechnology Division, Institute of Himalayan
Bioresource Technology, Palampur (HP), India
Plants are the source of a range of bioactive metabolites, which are used for good health, to treat various ailments and as important constituents of colours, dyes, flavours and fragrances. Most of these metabolites are synthesised through flavonoid (FL), and terpenoid [2-C-methyl-Derythritol 4-phosphate (MEP) and mevalonate (MVA)] pathways. One of the critical issues is to unravel the molecular control of synthesis of these compounds in the relevant plant systems. Catehcins, shikonins and picrosides are medicinally important metabolites synthesized in tea ( Camellia sinensis ), arnebia ( Arnebia euchroma ), and picrorhiza ( Picrorhiza kurrooa ), respectively. Using a range of genomic and biochemical tools, molecular regulation of FL pathway in tea, MVA pathway in arnebia, and MVA and MEP pathways in picrorhiza were deciphered and analyzed. Use of germplasm with varying levels of metabolites and environmental cue mediated modulation of metabolite content, followed by transcriptome analysis and functional validation of genes led to important outcomes as follows: (1) deciphered various genes involved in the biosynthetic pathways, (2) regulatory genes of the pathway were identified, (2) within a plant system, crosstalk existed between the connecting pathways, (3) early genes were important in determining the final product content, (4) environmental cues determined metabolite level through gene regulation; identified plant system specific critical cues, (5) product mediated feedback inhibition was evident. Using clues from the data generated, heterologous expression of important genes of FL pathway produced flavonoides in alternative
92 system and also fermentor-based shikonins production system was developed. Plants of medicinal value have been analyzed in detail to define pathways responsible for the synthesis of compounds of interest to human health. The symposium brings together a diverse range of interests dealing with gene networks involved in the production of compounds of medicinal interest as well as those involved in detoxifying environmentally harmful pollutants.
Improving the micronutrient content of the white rice
1 grain
Johnson, A 1
The University of Melbourne, Australia
The 'Green Revolution' that occurred between the 1940s and 1970s saw an enormous increase in agricultural productivity worldwide. The adoption of semi-dwarf and hybrid cereal varieties, as well as improved irrigation, fertilizer and pesticide practices, greatly increased food production in developing countries of the world.
However, not all changes brought about by the Green
Revolution were desirable. Cereal crops such as rice and wheat, while high in calories, do not contain significant quantities of nutritionally important micronutrients such as iron (Fe), zinc (Zn) and Vitamin A. As|increasing numbers of people have come to rely on cereal crops to meet their energy needs we have seen a reduction in the number of people suffering from caloric hunger but a massive increase, in fact billions of people, suffering from micronutrient hunger – a nutritional disorder often referred to as 'hidden hunger' because the effects, while devastating, are not always apparent at first glance. The effects of climate change are likely to further compound this problem. In particular, rising atmospheric carbon dioxide concentrations are predicted to decrease the protein concentration of most crops. Additionally, the concentrations of many essential elements in food crops, such as Fe and Zn, are predicted to decrease as a result of increased atmospheric carbon dioxide. These findings suggest that climate change may further compound severe micronutrient malnutrition problems already affecting billions of people throughout the world and highlight a need for the development of more nutritious crops. Because rice ( Oryza sativa L.) serves as the primary source of calories for more than half of the world's population, we are working to improve the micronutrient content of the white rice grain.
Nicotiana mine (NA) is a natural metal cation chelator that plays important roles in metal acquisition and transport in all plants. To generate rice lines with increased levels of Nicotiana mine, the three rice
Nicotiana mine synthase (NAS) genes – OsNAS1,
OsNAS2, and OsNAS3 – were constitutively overexpressed using the 35S cauliflower mosaic promoter.
Elemental analysis showed that the resulting rice lines contain three-fold more Fe and two-fold more Zn in white rice. Seed Nicotiana mine content is tightly correlated with Fe and Zn content, providing strong evidence that increased Nicotiana mine levels are responsible for the observed increased loading of Fe and
Zn into white rice grain. The levels of Fe in these biofortified rice varieties could meet the recommended daily allowance of Fe in a daily serving of rice. We anticipate that biofortification of all major food staples

will take on an increasingly vital role as the human population continues to grow under changing climatic conditions.
Lupin–wheat bread protein: modification of the bread matrix for improved health attributes
Shahidul, I 1
R 1
, Ma, W 1 , Yan, G 1 , Gao, L 1 , Yan, Y 1 , Appels,
1 Murdoch University, Australia
Nutritional qualities of lupin suggest bread rich in lupin has the potential to provide a number of health benefits.
Mixing of lupin flour to wheat has major effects on bread properties including the extractability of proteins that is crucial to define the ultimate attributes of food. Present study investigated the lupin and wheat proteome dynamics as affected by the baking of lupin–wheat bread by using 2-D electrophoresis and direct mass spectrometry. Many of the proteins from both lupin and wheat remained unchanged in baked lupin–wheat bread, while the others were incorporated into the bread matrix and could not be extracted. Most of the
α
-conglutins could be readily extracted from the lupin–wheat bread even at milder extraction. In contrast, most of the
β
conglutins lost extractability. The structural attribute that correlates with this difference in behaviour is the greater in thermal stability of
α
-conglutins relative to
β
conglutins. Most of the beta and gamma conglutins showed relatively higher peptide sequence coverage of corresponding matched proteins compare to alpha conglutin due to lack of information regarding this group of protein in the database.
Diversity of seed storage protein among narrowleafed lupin cultivars (
Lupinus angustifolius
L.) with reference to contributing to health
Shahidul, I
G 1
1 , Ma, W 1 , Appels, R
1 Murdoch University, Australia
1 , Buirchell, BJ 1 , Yan,
Narrow-leafed lupin (NLL) is one of the major legume crops in Australian farming system which is largely used as animal feed. A number of modern cultivars have been developed through breeding making NLL possible to be consumed as human food. Significant health benefits have been recognised when NLL seeds are consumed.
This study characterised protein polymorphism among the 25 Australian cultivars through mass spectrometric
(MALDI-TOF) with the aim of developing molecular breeding strategies to improve protein quality and content. Altogether, 364 seed storage proteins were identified by the MALDI-TOF profiling and fifty proteins were cultivar specific. Nine common proteins
(present in all cultivars) and 61 rare proteins (present in
2–3 cultivars only) were recorded. Phylogenic analysis based on the protein profile clustered the cultivars into 2 major groups and 5 subgroups which are generally supported by the pedigree information. Small number
(2.4%) of common proteins among the cultivars suggested a high level of genetic diversity in seed storage protein of NLL.
Interactions between regulatory pathways for abiotic stress tolerance in cereals
Dolferus, R 1 , Ji, X 1
1 CSIRO Plant Industry, Canberra, Australia
The reproductive stage is very sensitive to a variety of abiotic stresses (drought, cold, heat and salinity).
Adverse environmental conditions during flowering lead to loss in grain number and high levels of sterility in the world's most important staple crops. Experiments under controlled environment conditions using precisely controlled applications of stress periods have indicated that the young microspore stage of pollen development is the weakest link in the reproductive cycle. Both cold and drought stress cause abortion of pollen development in rice and wheat. This is associated with stress-induced repression of sugar transport in the anthers. Germplasm tolerant to cold and drought stress has been identified in rice and wheat respectively and it was shown that tolerance to cold and drought stress is associated with maintenance of sink strength. Cold and drought tolerant germplasm in rice and wheat is able to maintain lower
ABA levels in the spike during stress conditions. ABA represses cell wall invertase expression in anthers and transgenic rice plants over-expressing ABA 8'hydroxylase under the control of a strong tapetumspecific promoter results in significantly improved coldtolerance. The control of ABA homeostasis is therefore critical for maintaining pollen fertility and grain number during abiotic stress conditions. The high degree of similarity between cold-induced pollen sterility in rice and drought-induced sterility in wheat suggests that a common regulatory mechanism is involved in tolerance to both stresses and possibly other abiotic stresses.
Drought-tolerant wheat germplasm is also very tolerant to shading stress, suggesting that different stress tolerance pathways (including response to light intensity and quality) converge at a common regulatory node that controls reproductive development. Preliminary data suggest that auxins may play an important role in controlling ABA homeostasis and pollen development.
Molecular and genomics approaches, combined with
QTL mapping, will be used to identify genes responsible for reproductive-stage abiotic stress tolerance in cereals.
1
Water deficit and the anther tapetum
Parish, R 1
Botany Dept, La Trobe University, Melbourne Campus,
Victoria, Australia
Pollen fertility is one of the limiting factors in crops such as rice and wheat. Interference with normal tapetal development leads to pollen abortion and tapetal cells in many crops are sensitive to environmental stress.
Knowledge of the mechanisms underlying this sensitivity may facilitate the development of molecular approaches aimed at increasing the stress tolerance of the tapetum.
There is now considerable information available about the genes regulating tapetal development in Arabidopsis
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and rice, many of which are homologs. The timing of programmed cell death is critical for pollen development and any interference results in loss of pollen fecundity.
One of the transcription factors we are studying regulates the timing of tapetal PCD by controlling the transcription of an aspartic protease gene.
Stem carbohydrate remobilization as a key variable for wheat undergoing terminal drought
Zhang, J 1 , Dell, B 1 , Appels, R 1
1 Murdoch University, Australia
Terminal drought is an increasing risk for wheat production in many parts of the world, especially those with mediterranean type climate. The improvement of drought tolerance has been identified as a research priority. Under terminal drought, the impact of stem carbohydrate remobilization makes significant contributions to grain filling. Our study showed that the stem water soluble carbohydrate (WSC, main components fructan) level is not, on its own, a reliable criterion for the drought tolerance, the performance of the key enzymes involved in the WSC remobilization, which varied among different varieties, were critical in the terminal drought response. The genes involved in the mobilization of stem WSC, 1-FEHs and 6-FEHs (fructan exohydrolases), have been studied in detail to define their role in contributing to water deficit tolerance. The study includes the isolation of mutations where genes of interest have been deleted.
The cell wall invertase (IVR1) gene from wheat
Webster, H
P 1
1 , Fosu-Nyarko, J 1
, Bellgard, M 1 , Appels, R 1
, Keeble, G 1 , Moolhuijzen,
1 Centre for Comparative Genomics, Murdoch University,
Perth, Australia
Genetic analyses of controlled crosses in wheat have identified a region on chromosome 3B (short arm) that controls variation in water stress tolerance. The region, near the Sr2 resistance locus, is covered by a series of well studied BAC clones that are being sequenced as part of a large 3B sequencing program in France (led by
Catherine Feuillet, INRA, and the International Wheat
Genome Sequencing Consortium). A 1Mb region from this part of the wheat genome (ctg506) was assembled based on sequence data from overlapping BAC clones.
The sequencing of the BAC clones combined standard
Sanger sequencing (5-8 x coverage) and Solexa/Illumina short read sequencing. Annotation of the sequence identified the IVR1 gene considered to be important in conferring drought and frost tolerance to wheat when the stress occurs early in head development. Alignment to the rice and Brachypodium genome sequences indicated very little synteny and detailed analyses provided a sequence based model accounting for the evolutionary instability in this region of the genome.
Molecular dissection of genotypic variation in accumulation of water soluble carbohydrates in wheat
McIntyre, CL 1
D 1 , Dreccer, MF
Xue, GP 1
, Casu, RE 1 , Rattey, A 1 , Kam, J
1 , Van Herwaarden, AF 1
1 , Seung,
, Shorter, R 1 ,
1 CSIRO Plant Industry, QBP, St Lucia, QLD, Australia
Water soluble carbohydrates (WSC) stored in the wheat stem are an important contributor to wheat grain yield and grain size in all environments but especially when water is limiting. We are using both genetic and genomic approaches to determine the genetic basis of this important drought-adaptive trait. We have identified quantitative trait loci (QTL) for WSC in several wheat populations with individual QTL explaining small amounts of phenotypic variation. We have evaluated variation in gene expression in high and low WSC wheat progeny lines from a wheat population and found significant differences in expression of gene from different gene categories. For example, high WSC progeny lines have higher levels of expression of genes involved in carbohydrate metabolism and lower levels of expression of genes involved in cell wall and amino acid metabolism than low WSC lines. Genetic mapping reveals that several candidate genes co-locate with QTL for WSC. In addition, we have identified expression QTL
(eQTL) for candidate genes that co-locate with WSC
QTL; co-location of the genes and eQTL with WSC QTL make these genes stronger candidate genes for the WSC trait.
Generating plant reference genomes
Xun, X 1 , Jun BGI, W 1
1 BGI-Shenzhen, China
For those important crops, only one reference is insufficient to capture (for example) the more than 300 functional novel genes that differentiate between maize inbred lines. The variations between two different strains were as significant as the difference between human and chimpanzee. To understand the genomes, use it as a reference or find markers, the old strategy by only 'resequencing' is not enough for such variety species, especially for plant species. In potato genome annotation, the transcriptomes from more than 50 tissues or stages were mainly used and demonstrated the power of this approach for annotation especially where genes have long introns. Due to the complexity of some plant genomes such as highly heterogeneity (such as potato), polyploidy (wheat, cotton) and highly repetitive sequence
(maize, wheat), finishing a reference genome is a major challenge. To solve these problems, the BGI assembly now utilizes the BAC by BAC approach purely by Hiseq
2000 data, making it high quality and cost efficiency.
This illustrated by showing finished variation maps for some important crops, including maize, soybean and rice.
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A multifaceted approach to enhance switchgrass as a bioenergy crop
Devos, K 1
1 University of Georgia, Athens, USA
Switchgrass is a grass native to the prairies of the United
States. In the 1990s, it was earmarked as a crop with a potential for cellulosic bioenergy production because of its ability to produce high yields under marginal inputs.
However, it is only in the past few years that genetic research on switchgrass has started in earnest.
Switchgrass belongs to the subfamily Panicoideae, tribe
Paniceae. Lowland switchgrass is usually tetraploid
(2n=4x=36), with a 1C content of 1350 Mb. It is also an obligate outcrosser, which greatly complicates genetic analyses. We therefore are using a multifaceted approach to create the genetic, genomic and breeding resources that are required for building a successful switchgrass biofuel production program. A comprehensive genetic map using SSR and DArT markers has been constructed in a cross between Alamo AP13, a lowland clone, and
Summer VS16, an upland clone. The map is used for both trait mapping and to enhance our understanding of the organization of the switchgrass genome relative to that of its sequenced relative, foxtail millet ( Setaria italica ). A fosmid library with ~6X coverage has been generated and >30 fosmids containing genes involved in switchgrass cell wall synthesis/composition have been selected, sequenced and annotated. More than seven million ESTs have been sequenced from AP13 and VS16 and placed in a searchable database. The EST data will be used for the identification of SNPs to further enhance the genetic map. We are also assessing the diversity that is present in switchgrass germplasm. A panel of 384 switchgrass genotypes has been genotyped with SSR markers to assess the population structure and is currently being phenotyped for the presence of natural variation in components that affect recalcitrance to ethanol production, biomass yield and disease resistance.
This information is fed into the breeding program to enhance the development of switchgrass cultivars with desirable cell wall composition and increased yield for commercialization. In summary, our project provides the essential resources to conduct structural and functional analyses in switchgrass.
Foxtail millet as a model for biofuel feedstocks
Bennetzen, J 1
1 University of Georgia, USA
Grasses of the genus Setaria , including the domesticated
Foxtail millet ( S. italica ) and one of the world’s most problematic weeds, Green foxtail ( S. viridis ), have many of the properties that are essential for the development of a model system for basic and applied research. In addition, Setaria is closely related to several candidate biofuel feedstocks, including Switchgrass ( Panicum virgatum ), Napier grass ( Pennisetum purpureum ) and
Pearl millet ( Pennisetum glaucum ). S. viridis is the apparent progenitor of Foxtail millet and is one of the most widely distributed grasses on the planet, including its common occurrence as a weed wherever wheat is grown. Both S. italica and S. viridis are true diploids, with some Green foxtail accessions that mature at less than 20 cm and produce mature seed in as little as 8 weeks after planting. The genome of a Foxtail millet cultivar, Yugu1, has now been fully sequenced (8.4X) by the Sanger approach, along with 454 sequencing of numerous EST libraries. Annotation is underway, as is the analysis of one deeply sequenced (by an Illumina approach) S. viridis accession, A10, and eight other S. viridis accessions sequenced at a lower redundancy.
Projects are underway to assess diversity, population structure, plant morphology, biomass productivity, herbicide tolerance, cell wall composition, and several other properties in a range of germplasm and in mapping populations. Results of these studies, especially the comparison to other grasses and to the Setaria genetic map, will be discussed.
1
The amaizing genomes of maize
Chia, J-M 1
Cold Spring Harbor Laboratory, USA
The first high quality reference sequence of Zea mays
(B73) provided insight into its origin from ancient tetraploidy and is propelling research to describe the tremendous diversity of this species. The reference coupled with the enabling sequencing technologies allow us to capture genomic, epigenetic and expression variation. I will discuss the results of several collaborative projects aimed at mapping variation and understanding its sources and consequences. The Maize
Diversity Project has developed a high-resolution variation map that integrates millions of segregating
SNPs, indels, and copy number variations (CNV) obtained from more than 100 different maize backgrounds. This panel included domesticated improvement lines, early-domestication landraces, and pre-domestication teosinte, which were subjected to resequencing at extremely deep coverage (500X). This sequence depth and sample breadth enabled us to identify regions of genetic fluidity versus rigidity in the face of selection pressures. A separate study focused specifically
CNV and presence–absence variation (PAV) within a large panel of diverse inbreds and teosinte lines. These polymorphisms were found to be more prevalent among particular gene-classes, including members of tandem arrays, large families, recently duplicated, and recently evolved genes. These results suggest that many gene losses are buffered by redundant functions encoded elsewhere in the genome.
Updated genome information of
Lotus japonicus
and status of legume comparative genomics
Sato, S 1
1 Kazusa DNA Research Institute, Japan
In order to investigate the whole genetic system of legume species, we have been analyzing the genome of a model legume, Lotus japonicus . In 2008, we have released the 315 Mb sequences determined by combination of two independent approaches; clone-byclone sequencing of large insert clones and random sequencing of selected genomic regions. We continued our efforts on sequencing of genome clones, and the
95

sequence information on 460 clones has been accumulated. Thus, we have updated the sequence information by adding these clone sequences. As a result, the total length of the pseudomolecule, which means genome regions covered by genome clone sequences, was increased from 167 Mbp to 201 Mbp, and the total size of finished (phase 3) sequences was increased from
97 Mbp to 139 Mbp. The detailed information on the updated sequences and annotation is available through our web database, 'miyakogusa.jp'
(http://kazusa.or.jp/lotus/). Using the updated genome sequence and predicted gene information, we have carried out comparative genome analysis against
Medicago truncatula and soybean. As a result, high level of syntenic relationship was identified among these legume species. We extended our efforts on comparative genome analysis in legume species to red clover
( Trifolium pratense ) and white clover ( T. repens ) using their genetic linkage maps created by EST-derived markers, and identified significant level of syntenic relations against L. japonicus, M. truncatula and soybean. Toward the completion of L. japonicus genome sequencing, we started an additional approach by using the sequence data from the second generation sequencers.
By integrating paired end sequences generated from 454 and Solexa sequencers, scaffolding and gap filling of current contigs are on going. Status of this integration approach will also be presented.
Structure–function studies in the wheat genome
Keeble, G 1
Bellgard, M 1
, Black, M 1
, Appels, R 1
, Moolhuijzen, P 1 , Barerro, R 1 ,
1 Centre for Comparative Genomics, Murdoch University,
Perth, Australia
Two separate 1Mb regions of the wheat genome comprising extensive stretches of repetitive sequences were assembled based on sequence data from overlapping BAC clones located in a section of the chromosome 3B of particular interest with regards to disease resistance. The sequencing of the 24 BAC clones combined standard Sanger sequencing (5-8 x coverage) and Solexa/Illumina short read sequencing (>300 x coverage). In addition, available BAC end and 454-based sequences were utilized to provide a good quality reference sequence that was annotated with a particular focus on the repetitive elements. Alignment to the rice and Brachypodium genome sequences identified a repetitive locus that is conserved between these genomes and the wheat genome. The conserved repetitive protein kinase like genes at this conserved locus showed a good alignment to the bacterial effector protein AvrPto
(Weiman et al. 2007).
1
A systems approach to defining C
4
networks in the grasses
Brutnell, TP 1
C
Boyce Thompson Institute, USA
4
photosynthesis has evolved at least 60 times in the angiosperms and at least 17 times in the grasses, leading to some of the most productive food, feed, and bioenergy crops on the planet. Yet, we know very little of the regulatory networks that drive this process in any major crop plant. I will describe our efforts to use maize and
Setaria viridis as models for a systems biology approach to understanding regulatory networks driving C
4 differentiation. We have recently analyzed the maize leaf transcriptome using Illumina sequencing (RNAseq) to define gene structure and to quantify transcript abundance along a calibrated leaf developmental gradient and in mature bundle sheath and mesophyll cells.
Complementing this analysis, detailed histological, proteomic and metabolite datasets have been collected along the same gradient and analyzed by several collaborators. We are now developing methods to integrate these diverse datasets and I will discuss our current strategies and findings. To provide community access to this data, we implemented Gbrowse and an electronic fluorescent pictograph browser to display and query the datasets. Finally, I will describe our recent efforts to develop Setaria viridis as a model system for dissecting C
4
gene expression.
Plant epigenetics : stories from beyond the double helix
1
Rival, A
F 1
1 , Jaligot, E 1
, Adler, S 1
, Beule, T 1
, Debladis, E 1
, Richaud, F 1
, Ilbert, P 1
, Aberlenc,
, Finnegan, J 2
UMR DIADE. IRD, Cirad BioS, Montpellier, France;
2 CSIRO Plant Industry, Canberra, Australia
Epigenetics is the study of heritable changes in gene function that occur without a change in the DNA sequence. In recent years, this field has attracted much attention as more epigenetic controls of gene activities are being discovered. Such controls involve a complex interplay of DNA methylation, histone modifications, and RNA-mediated pathways from non-coding RNAs, notably silencing RNA (siRNA) and microRNA
(miRNA). In plants, although epigenetic mechanisms help to protect cells from parasitic elements, this defence can complicate the genetic engineering process through transcriptional gene silencing. Furthermore, these phenomena have economic relevance, for example, in somaclonal variation: a genetic and phenotypic variation among clonally propagated plants from a single donor genotype. The loss of phenotypic fidelity is now a major impediment to the development of large scale propagation of plants through in vitro processes such as somatic embryogenesis. Examples of aberrant phenotypes in regenerated plants include abnormal leaf structures and variant floral morphology, both organs
96

being of paramount importance for applications in horticulture and/or agriculture. Changes in DNA methylation have been hypothesised playing a key role in the mechanism underlying tissue-culture induced changes. Indeed, studies of both global methylation levels and the methylation of specific sites show that variation in DNA methylation occurs frequently during growth in tissue culture. In vitro plant regeneration, like somatic embryogenesis, bypasses the normal developmental process of fertilisation and plant development, thus potentially resulting in the instability of epigenetic patterns. The large-scale clonal propagation of oil palm ( Elaeis guineensis Jacq.) is being hampered by the occurrence of the mantled somaclonal variation.
Indeed, this abnormality which presents a homeotic-like conversion of male floral organs into carpelloid structures, hampers oil production since the supernumerary female organs are either sterile or produce fruits with poor oil yields. Beyond its primary interest in the search for discriminating markers against an economically crippling phenotype, the study of the mantled abnormality also provides a unique opportunity to investigate the regulation of reproductive development in a perennial tropical plant. The present interest on food and energy security as well as the concerns raised by the possibility of climate change further stress the need for a global comprehension of how crop plants react to their fluctuating environment and how their productivity can be affected.
Comparative genomics in clovers (
Trifolium
spp.) using
Medicago truncatula
Gaertn. as a reference for candidate genes controlling agronomic traits
Ghamkhar, K 1
Ryan, MH 4
, Isobe, S
, Snowball, R 3
2 , Nichols, P
, Sato, S 2
3 , Faithful, T
, Appels, R 5
1 ,
1 University of Western Australia, Australia;
DNA Research Institute, Japan;
2 Kazusa
3 Dept of Agriculture &
Food Western Australia, Australia; 4 School of Plant
Biology, Faculty of Natural and Agricultural Sciences,
The University of Western Australia, Australia; 5 Centre for Comparative Genomics, Murdoch University,
Australia
Background: Molecular markers and genetic linkage maps are pre-requisites for molecular breeding, but no genetic map is currently available for subterranean clover
( Trifolium subterraneum L.), a diploid and inbreeding pasture legume. A total of 2,646 microsatellite or simple sequence repeat (SSR) markers either available in the public domain or unpublished obtained from Kazusa
DNA Research Institute, were screened on four genotypes (Denmark, DGI007, Woogenellup and Daliak) that are parents of two F2 mapping populations. Results:
A total of 343 SSR loci could be mapped into 8 linkage groups with 6–31 loci per linkage group and 27 SSRs shared between the two populations. Phenotyping data obtained for flowering time, isoflavonoid content
(formononetin, genistein, biochanin A), hardseedness, leaf marks, calyx pigmentation and hairiness of stem were analyzed together with genotyping data. For each trait, 1–3 QTLs were identified; the phenotypic variation explained by these QTLs was in the range of 5.5–59.8%.
In addition, alignment of linkage groups was possible with the genetic maps of red clover ( Trifolium pratense
L.), white clover ( Trifolium repens L.)and Medicago truncatula Gaertn. using shared loci. Segmental duplications were identified within subterranean clover genome and between the legume species. Candidate genes for isoflavone content were identified.
Conclusions: The present study reports on comparative genomics among three clover species as well as
Medicago truncatula following the construction of the first genetic map for subterranean clover. It demonstrates the utility of a comparative genomics approach for molecular mapping of QTL controlling seed hardness, flowering time and specifically isoflavone content traits, as well as establishing relationships between subterranean clover and genomes of red clover, white clover and model legume M. truncatula . The comparative mapping carried out demonstrated the value of the map to identify candidate genes controlling important traits such as isoflavone content.
1
Sucrose accumulation in stems of sweet
Sorghum bicolor
Byrt, CS 1 , Milne, R 1
University of Newcastle, Australia
Stem juice sugar content and juice volume were measured in 135 diverse Sorghum bicolor ecotypes.
Pressed juice contained up to 25% sucrose and juice volumes of up to 500 mL per plant were measured. The higher yielding ecotypes produced the equivalent of 10
000 Kg of sugar per ha. Sucrose transporter (SUT) proteins play an important role in the accumulation of sucrose in stems of sweet Sorghum bicolor varieties and six putative SUTs have been identified in Sorghum. The diversity in stem sugar yields and the availability of the full genome sequence makes Sorghum an ideal C
4 monocotyledonous species in which to study the role of
SUTs. Differing patterns of expression were observed when transcript levels of all six SUTs were compared in a low and high sugar line of Sorghum at two developmental stages, immediately prior to anthesis and at the stage of peak stem sugar content. Complementary
DNA (cDNA) of all six SUTs was isolated from a low and a high sugar line and the predicted protein sequences of a number of the SUTs were found to differ between the two lines. A number of SUTs of interest were expressed in Saccharomyces cerevisiae and functional transport characteristics compared. The cellular location of the SUTs was also investigated, using two antibodies.
The PEP2 antisera, raised against a highly conserved region of sucrose transporters between loops 2 and 3 corresponding to amino acids 87 to 106 of the potato
StSUT1, was considered likely to bind to five of the six
SbSUTs, based on peptide sequence homology. The second antibody raised against ShSUT4, was expected to bind specifically to the remaining Sorghum SUT,
SbSUT4. SUT transcript levels, protein localisation and
SUT transport rates will be discussed.
Cross-species characterization of APOSTART for unveiling its role in apomixis
Marconi, G 1
3
, Galla, G 2
, Barcaccia, G 2
, Conner, J 3 , Raggi, L
, Falcinelli, M 1
1 , Ozias-
, Albertini, E 1
1
Akins, P
Applied Biology Dept, University of Perugia, Perugia,
Italy; 2 Environmental Agronomy and Crop Science Dept,
97

University of Padova, Legnaro, Italy; 3 Dept of
Horticulture, University of Georgia Tifton Campus,
Tifton, USA; 4 Leibniz Institut für Pflanzengenetik
Kulturpflanzenforschung, Gatersleben, Germany
Although most desirable crop traits are polygenic, no plant breeding tools exist which allow the efficient fixation of multigenic traits over successive generations.
Among several reproductive system-related strategies for fixation of desirable agronomic traits, one of the best choices is apomixis (i.e. clonal seed production), which would enable the instantaneous fixation of the complete genome of the best plants. Moreover, when coupled with male-sterility systems, apomictic reproduction (e.g. autonomous systems, with no need for male contribution) could help in addressing issues related to transgene escape from GM crops to organic or conventional crops, and thereby allow for better coexistence systems in
Europe. The development of apomixis technology is expected to have a revolutionary impact on food and agriculture production by reducing cost and breeding time, avoiding complications typical of sexual reproduction (i.e. incompatibility barriers) and of vegetative propagation (e.g. viral transfer). The financial and economic impacts of the development of apomixis technology and its application to major crops are considerable. It is estimated that apomixis technology in the production of hybrid rice alone could provide benefits exceeding 1,800 million Euros per annum. The development of apomixis technology in agriculture will require a deeper knowledge of the mechanisms regulating reproductive development in plants. We have isolated one gene, which was termed APOSTART because of its START domain and its putative involvement in apomixis. START was named after the discovery of the StAR gene involved in human congenital lipoid adrenal hyperplasia whose clinical phenotype includes the onset of profound adrenocortical insufficiency shortly after birth and male pseudohermaphroditism. Our results demonstrate that some APOSTART members are expressed exclusively in inflorescences of Poa pratensis . In situ hybridization analyss revealed that APOSTART is expressed during both male and female meiosis in all micro- and megaspores. Strong signals were recorded up to the mature embryo sac stage. Overall data suggest that
APOSTART may be related to the programmed cell death that is involved in the non-functional megaspore and nucellar cell degeneration events that permit enlargement of maturing embryo sacs. To confirm its involvement in apomixis, we have isolated APOSTART members from two other aposporic species: Pennisetum squamulatum and Hypericum perforatum .
Uncovering genome features of crops with high throughput sequencing
Michael, T 1
1 Monsanto, St Louis, USA
High throughput sequencing has fundamentally changed the way that we do genome discovery. The time that it takes from sample to genome can be days for hundreds of dollars, compared to just five years ago when it would have taken months and cost hundreds of thousands.
Moreover, high throughput sequencing has enabled yet
98 another interesting turn of events: sequencing has morphed into functional genomics. The ability to generate trillions of sequences at a time turned sequencing into a genomic discovery tool for such applications as RNA expression analysis, metagenomics,
DNA binding studies, histone and nucleosome mapping, and DNA methylation detection. Sequencing technologies have matured into platforms for molecular genome phenotyping, which changes the paradigm and scale of understanding the link between genotype and phenotype. This fundamental shift has fueled the innovation of now emerging genome tools such as real time single molecule detection (Pacific Biosciences RS), and post-optic sequencing (ION Torrent/PGM).
However, these genome machines cannot deliver biotech products by themselves. New bioinformatic and analysis tools are emerging to derive knowledge from the growing mountains of data. I will talk about how high throughput sequencing is enabling genome discovery in crops.
Comparative mapping in
Eucalyptus genomes
Vaillancourt, RE
AA 2 , Faria, DA 3
1 , Hudson, CJ 1
, Grattapaglia, D 4
, Kumar, AR 2
, Kilian, A 5
, Myburg,
1 University of Tasmania & CRC for Forestry, Hobart,
Australia;
Africa; 3
2 University of Pretoria, Pretoria, South
EMBRAPA, Brasilia, Brazil;
Catolica de Brasília, Brasilia, Brazil; 5
4 Universidade
Diversity Arrays
Technology Pty Ltd, Canberra, Australia
The completion of the Eucalyptus grandis reference genome sequence reinforces the need for a better understanding of genome homology in the genus.
Genome homology can occur at multiple levels from the location of loci on homologous linkage groups (synteny), the congruent ordering of loci on chromosomes
(colinearity), to conservation of DNA sequence. It also involves understanding the extent to which gene polymorphisms (SNP) and their effects on the phenotype
(QTL) are shared across species. Differences in genome structure may explain variation in the fitness of hybrids at various taxonomic scales. In addition, sharing of QTL for genes of adaptive significance may explain cases of phenotypic convergence in the genus. From a practical point of view, this knowledge would help guide the transfer of molecular information across species and accelerate molecular breeding. Most commercially important eucalypts belong to the largest of the eucalypt subgenera: Symphyomyrtus . Despite nearly 50% variation in DNA content across these species, all appear to be diploid with 2n = 22. There is no evidence of chromosome rearrangements, although the chromosomes are so small that these would be difficult to detect microscopically. We conducted comparative genomic studies between Symphyomyrtus species using highdensity linkage maps based predominantly on DArT markers in five large E. globulus (section Maidenaria )

families and a double pseudo-backcross mapping family involving the species E. urophylla and E. grandis (both in section Latoangulatae ). Comparison within sections
( E. globulus versus E. globulus or E. grandis versus E. urophylla ) found very high synteny and colinearity.
While the degree of co-linearity between homologous chromosomes in the inter-sectional comparison ( E. globulus consensus versus the E. grandis – E. urophylla consensus) was similar to that found in the within section comparisons, the percentage of non-syntenic markers increased more than four-fold. While some of these nonsyntenic markers may be mapping errors, there is evidence for three very small rearrangements, which are probably translocations. This is because the map position of these non-syntenic markers were consistent between different maps of E . globulus and consistent between the
E. grandis map and the genome sequence. There is also evidence that some QTL for developmental, defensive chemistry, physical and chemical wood property traits are shared between species. Our results argue for high genome homology overall and high transferability of the
E. grandis genome sequence information and marker/trait associations across the subgenus
Symphyomyrtus . This high transferability will facilitate the identification of genes underlying phenotypic variation in this ecologically and economically important lineage of tree species.
Molecular adaptation to water availability in
Eucalyptus
Bragg, JG 1 , McEvoy, R 1 , Dillon, SK 1 , Thumma, BR 1
1
Southerton, SG 1
CSIRO Plant Industry, Canberra, Australia
,
The genus Eucalyptus radiated during a time of great environmental change on the Australian land mass, and extant eucalypts populate diverse ecosystems across
Australia. The genus therefore provides excellent opportunities for exploring the genomic basis of adaptation to a wide range of environmental conditions.
We are studying adaptation to water availability in
Eucalyptus camaldulensis and several other species. This research is being carried out at a range of different taxonomic and spatial scales (among species and among populations), using both candidate-gene and genomewide approaches. Our aim is to identify loci that exhibit signatures of natural selection in populations from environments that vary in water availability.
Using the
Eucalyptus genome to understand the evolution of plant secondary metabolites in the
Myrtaceae
Kulheim, C 1
W 1
, Webb, H 1 , Wallis, I 1 , Moran, G 1 , Foley,
1 Australian National University, Canberra, Australia
Eucalyptus trees (family Myrtaceae) are well known for their high foliar content of several classes of secondary metabolites and these have a strong effect on the feeding patterns of several species of marsupials and at least some insects. Best known are the essential oils, which are mostly a mixture of terpenoids but there are also significant concentrations of flavonoids and formylated phloroglucinol compounds. Quantitative and qualitative variation within and between species in all these groups is large with evidence that much of this variation is under strong genetic control (heritabilities of between 0.3 and
0.9 for all three groups of compounds). As well as being important ecologically, the terpenes in particular are valued as industrial and medicinal products and Australia supports a strong essential oil industry focused on
Eucalyptus and Melaleuca foliar oils. 1. The Eucalypt genome provides the opportunity to discover the genetic makeup of the biosynthetic pathways for secondary metabolites. We present data from pathways leading into the biosynthesis of terpenes and flavonoids. The homology of genes and gene families were investigated and compared to a variety of model species including poplar ( Populus trichocarpa ), grape ( Vitis vinifera ) and apple ( Malus x domesticus ). For example the gene family that commits the final step in the terpene biosynthesis
(terpene synthase family) has 120 members in the genome of Eucalyptus grandis , compared to 44 and 99 in poplar and grape, respectively. 2. Levels of genetic variation within and between species of the Myrtaceae were compared on the basis of multiple loci from the secondary metabolite biosynthetic pathways. Eleven loci were amplified from 110 individuals of Eucalyptus globulus and seven from 400 individuals of Melaleuca alternifolia . These loci were barcoded for each individual and sequenced on a next-generation sequencing platform.
This allowed to discover and genotype 100s of polymorphisms instantaneously. Population genetics parameters were estimated and compared to other
Eucalyptus species. The gene discovery was made possible through the Eucalypt genome. 3. Understanding the genetic basis of trait variation in quantitative traits may help breeders in the essential oil industry and also provide insights into ecological processes. We have characterized trait associations with polymorphisms from
Eucalyptus globulus , investigating 200 SNPs and roughly
70 traits including, terpenes, formylated phloroglucinol compounds, flavanoids and the functional effects of tannins. We discovered several significant trait associations between allelic variants in the chloroplastic
MEP pathway and monoterpenes and between the cytosolic MVA pathway and sesquiterpenes.
Systems genetics of wood formation in
Eucalyptus
Mizrachi, E 1 , Van Dyk, MM
CA 2 , Joubert, F 2 , Myburg, AA 1
1 , Kullan, ARK 1 , Hefer,
1 Dept of Genetics, Forestry and Agricultural
Biotechnology Institute (FABI), University of Pretoria,
Pretoria, South Africa; 2 Bioinformatics and
Computational Biology Unit, Dept of Biochemistry,
University of Pretoria, Pretoria, South Africa
Advances in Eucalyptus genomics in the past two years have enabled the use of new biological tools for studying fibre and lignocellulosic biomass properties in this genus, which represents one of the most important sources of cellulose globally for pulp, paper, and potentially biofuels. In collaboration with industry partners, we have produced a high quality interspecific mapping pedigree of Eucalyptus grandis and E. urophylla , using the same
F1 hybrid parent in a two-way pseudobackcross, which has produced more than 1000 F2 progeny. These display high levels of genomic and phenotypic segregation, and
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provide a powerful statistical framework for associating genes and markers with multiple traits in this population.
The availability of a high density genetic map for this population constructed using SSR and DArT markers, which have also been linked to the E. grandis reference genome sequence (JGI, V1.0), has allowed the construction of a high confidence framework map for
QTL mapping. In addition to profiling phenotypic traits of interest in over 300 individuals from each backcross, we have in collaboration with ORNL profiled 83 metabolites in the developing xylem of 192 individuals from one of the backcrosses and sequenced the developing xylem transcriptomes of the same 192 F2 individuals using Illumina RNA-Seq, for expression QTL
(eQTL) and metabolite QTL (mQTL) mapping purposes.
We present our strategy for data generation and integration, as well as the applications of Eucalyptus
Systems Genetics in reconstructing transcriptional networks and identifying pathways, candidate genes and regulatory sequences that influence wood and fibre properties of relevance to commercial end uses of wood such as pulp and paper production.
Eucalypt comparative genomics identifies model genes to study the effect of domestication or range depletion on adaptive variation
, Henry, R 3 , Henson, M 2 , Sexton, T 1 , Thomas, D 2
Shepherd, M 1
1 Southern Cross University, CRC for Forestry, Lismore,
2
3
Australia; Forests NSW, Coffs Harbour Jetty, Australia;
Present address: Queensland Alliance for Agriculture and Food Innovation, University of Queensland,
Brisbane, Australia
In plants, balancing selection may cause the retention of adaptive polymorphism over long time spans, with genes for biotic and abiotic stress responses among the best known of those affected. In the past, the inadvertent removal of polymorphism from domesticated populations otherwise maintained by balancing selection, has sometimes met with catastrophic consequences. e.g. Irish potato famine. Similarly, the unintentional threats from the loss of adaptive diversity and inbreeding to the fitness of natural populations that undergo reductions in size or fragmentation, are also widely documented. A better understanding of the prevalence of balancing selection, its modes of action (heterozygote advantage, frequencydependent selection and selection varying in time and space), and the genes it affects, might reveal new ways to manage and mitigate against the loss of adaptive variation during domestication or range depletion. The present study used comparative genome analysis to identify genes under balancing selection. Thirty four candidate genes for wood formation were screened for
SNP common to the Symphyomyrtus and Eucalyptus subgenera of the genus Eucalyptus . These transsubgeneric SNP likely derive from ancestral polymorphisms that are maintained in each lineage postspeciation by balancing selection. Genes containing clusters of trans-specific SNP are good models for monitoring the loss of adaptive variation because the drivers of selection in these genes may be universal in eucalypts. The study showed balancing selection was more common among candidate genes for wood formation than might be expected on average in a
100 genome, nonetheless, this high frequency was congruent with potential pleiotropic roles for these genes in biotic and abiotic stress responses. Strong evidence for balancing selection was found in ten genes where there was a prevalence of trans-subgeneric SNP clustered in functional gene regions. Two of these genes were from the pectin methylesterase gene family, key enzymes that catalyse the de-methyl-esterification of homogalacturonan, the major constituent of pectin. This gene family has pleiotropic functions in wood formation, seed dormancy, fruit ripening and stress responses.
Association testing found both genes also influenced physical and chemical wood phenotypes of interest for tree improvement. Adaptive variation in one of these genes was found to be maintained through a heterozygote advantage. These genes may be useful models for exploring the consequences of diversity reduction at genes subject to balancing selection.
Genomic impacts on eucalypt cultivation and
1 conservation
Wright, J 1
ArborGen Inc., Summerville, USA
The genus Eucalyptus includes species that comprise the most important hardwood plantation resource in the world. There are an estimated 100 eucalypt tree improvement programmes utilizing seedling or clonal propagation techniques with objectives as diverse as bioenergy to furniture grade lumber. These eucalypt improvement programmes are making great advances in productivity, industrial processing, improving rural livelihoods and job creation. However, perhaps only a handful of programmes are making use of recent advances in eucalypt genomics. A concerted effort to map the eucalypt genome would help immensely many current and future eucalypt tree improvement programmes as well as assist in gene conservation with native eucalypts in Australia, Indonesia and Papua New
Guinea. Genomic advances, aided by the sequencing of the euclaypt genome, could be used in marker aided selection, directed species hybridization, overcoming seed/clonal propagation difficulties, developing trees for climate changes as well as developing the eucalypts for more marginal sites including dry, marginal sites.
1
Genomics of wild crop relatives
Henry, R 1, 2
Queensland Alliance for Agriculture And Food
Innovation, University of Queensland, Australia; 2 Centre for Plant Science, Southern Cross University, Lismore,
Australia
Analysis of the genomes of wild crop relatives facilitates their use as genetic resources expanding the available gene pool to satisfy the needs of food security and adaptation to climate change. Genome, transcriptome and specific amplicon sequencing are all contributing to

accelerated knowledge of wild and domesticated plants and revealing new approaches to plant improvement.
Genome sequences have provided genetic explanations for traits selected by humans in domestication that would be deleterious to the survival of plants in the wild.
Analysis of nuclear, mitochondrial and plastid genomes can be achieved at the same time by sequencing of a conventional total DNA extract from the plant. The chloroplast is sequenced at high coverage in shot gun sequencing of total DNA. This provides a whole chloroplast genome sequence to use as a barcode for identifying the plant and phylogenetic analysis. Wild rice relatives from Australia and Asia have been sequenced to determine their relationships. The Australian species have been isolated from domestication in Asia and represent a valuable and diverse resource for use in rice improvement. Transcriptome sequencing allows discovery of biologically important or functionally useful genes in plant populations. Extensive sequencing of the transcriptomes of the developing wheat seed has identified genes that may determine nutritional value and food quality. Amplicon sequencing can be used to characterize wild plant populations. Variation in genes in wild populations found growing across environmental gradients can allow analysis of the genetic differences associated with adaptation to different environments and indicate ways to adapt agricultural plant varieties to climate change. This approach has shown that some grasses have a greater diversity in biotic defense loci in dryer environments. Large scale amplification and sequencing of candidate wood property genes from
Eucalypts has revealed evidence for balancing selection in these genes in wild populations. These insights explain the complex genetics of these traits important for plant performance in the wild and for human use.
Population genomics of non-model invasive species
Dlugosch, K 1
1 University of Arizona, USA
Introduced and invasive species are major threats to plant diversity, but they are also outstanding opportunities to study the ecology and evolution of populations colonizing novel environments. Large numbers of plants species are being introduced to new habitats around the globe, and most of these are non-model species.
Moreover, the genomic features that interest us in these systems necessarily reside in wild, outbread individuals:
How have allele frequencies changed across populations?
Where do we seee evidence of selection? What is the contribution of admixture and hybridization to population expansion? Genomics can allow us to answer these questions, but doing so, particularly without a full reference genome, presents a variety of challenges. I will report on several of these methodological issues for which we have released freely-available public software, including the editing and assembly of next-generation transcriptome (EST) sequence data, and the comparison of genetic variation among outbred (wild) individuals with incomplete genomic information. I will also present data from projects where I and my collaborators are using genomic approaches to investigate the causes and consequences of evolution in several species of invasive plants. One major explanatory hypothesis for invasions is that invaders have evolved along fitness trade-offs that are common to all plants, gaining competitive advantages at the expensive of other functions not selected in their new environment (such as defenses). An alternative
(though not mutually exclusive) hypothesis is that hybridization associated with species introductions generates novel genetic benefits for invaders. We are quantifying fitness levels in response to various biotic and abiotic environments, comparing expression level and genome-wide sequence variation among native and invading genotypes, and mapping the genetic basis of observed evolutionary increases in growth rate. Finally, I also report on a new project to quantify changes in mating system and elucidate the genetic basis of associated losses of inbreeding depression in invading populations.
Build a digital library of plant genomes
Yang, B 1
1 BGI, Shenzhen, China
The application of second-generation sequencing technology in genome sequencing has propelled us into a new era. Using highly parallel processing in microfluidic devices and imaging system, highly accurate sequencing results are obtained. Orders of magnitude larger amounts of data are generated at substantially decreased cost and effort. This has made it possible to sequence plants genomes not just for models. Up to the present, a number of plant genomes have been sequenced. BGI-Shenzhen has developed the cucumber (367Mb), Chinese cabbage
(500Mb) and potato (830Mb) genomes. Plant genomes are essential to understanding how plants function and how to develop desirable plant characteristics. For example, the sequenced cucumber genome affords insight into traits such as its sex expression, disease resistance, biosynthesis of cucurbitacin and ®fresh green odor. Due to the significant impact that knowledge of genome sequence has shown, the genomes of more plants are expected to be completed in the next few years. We are taking a genomics approach to build a digital library of plants. Obtaining the genome sequence of a plant will generate vast amounts of informative datasets and enable the comprehensive understanding of the plant. In particular, we are developing de novo sequencing of clade genomes, which will unveil the set of whole genome sequences in the same family or genus (core genome + variable genomes). Investigators of clade genomes will provide unique insight into mechanisms of the speciation process and functional elements of species that drive diversity in these genomes. Functional elements related to growth, architecture, productivity or resistance can be further applied in plant molecular breeding and benefit for farmers, the seed and processing industries, the environment and the customer in both the industrialized and the developing world.
Genomic footprints of hybridization
1
Barker, M 1
University of Arizona, USA
Natural hybridization has long fascinated botanists as a possible source of novelty and innovation, fueling a long running debate concerning the ultimate role of
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hybridization in plant evolution. One reason this debate has persisted since Darwin is that detecting ancient hybridization and introgression is difficult. Advances in genetic and phylogenetic tools have permitted the identification of a few homoploid hybrid species, but the time and effort required to make these diagnoses is not trivial. Here I present a novel and efficient method for identifying ancient hybridization and introgression by inferring intrologs (introgressed orthologs) in EST data sets. Using a combination of simulations and data from established hybrid species, I explore this new test and demonstrate the type of data required. Application of this novel test to the broadly available plant ESTs I find evidence of many other, previously undocumented examples of ancient hybridization. From this survey of plant ESTs, I also use this new method to discriminate allo- and autopolyploidy and provide an estimate of the relative prevalence of these two forms of polyploid speciation.
The milkweed genome project: sequencing and characterization of the
Asclepias syriaca
(Apocynaceae) genome based on illumina short read sequences
, Fishbein, M 2 , Cronn, RC 3 , Liston, A 1
1
Straub, SCK 1
2
Oregon State University, Corvallis, OR, USA;
3
Oklahoma State University, Stillwater, OK, USA;
USDA Forest Service, Forest Genetics, Pacific
Northwest Research Station, Corvallis, OR, USA
Milkweeds are important models for the evolutionary ecology of plant reproduction, plant hybridization, and plant-herbivore co-evolution. The common milkweed,
Asclepias syriaca L., is the most intensively studied milkweed, and as such is an attractive target for the development of genomic resources. The milkweed genome project aims to sequence the 835 Mbp genome of A. syriaca to a minimum of 25X coverage using the
Illumina platform for massively parallel DNA sequencing. With a lower coverage preliminary data set, most of the high copy fractions and some of the lowcopy fraction of the genome were assembled using a combination of reference guided and de novo assembly strategies. A complete 155 kbp plastome sequence was obtained, revealing that Asclepias has a typical asterid chloroplast genome, except that both accD and clpP are pseudogenes. More than 400 kbp of the mitochondrial genome was assembled and characterized for gene content. A 7.5 kbp nuclear ribosomal DNA cistron was assembled and ribosomal DNA copy number estimated to be approximately 1500 based on sequencing depth.
Low-copy nuclear genes were assembled through a comparative genomics approach utilizing data from other asterids, including Coffea, Solanum , and Capsicum . The repeats present in the milkweed genome were also surveyed. This scan revealed a high number of interspersed repeats associated with the long terminal repeat retrotransposons (e.g., Ty1-copia, Ty3-gypsy). In addition, numerous potential microsatellite loci were identified and evaluated. The markers obtained through this sequencing effort will be applied to milkweed genetic map development and to estimate phylogenetic relationships among the 135 species of Asclepias . This project will provide the first insights into the genomic basis of the incredible morphological and ecological
102 diversity observed among milkweeds. The genome will serve as a community resource for studies of plant-insect co-evolution, floral developmental genetics, chemical evolution, population genetics, and comparative genomics. The results to date also demonstrate the feasibility of extending genome sequencing to a wide range of organisms of evolutionary and ecological interest.
Amborella
: an evolutionary reference genome for plant biology
Soltis, PS 1 , Soltis, DE 1 , Barbazuk, WB 1
1
Walts, BM 1
University of Florida, USA
, Chamala, S 1 ,
The handful of angiosperm genomes that have been sequenced to date represent phylogenetically derived portions of the angiosperm Tree of Life and therefore provide few insights into the characteristics of the
'ancestral angiosperm'. A complete understanding of any gene or genomic feature requires a perspective based on evolutionary lineages, one that can be obtained through analysis of an appropriately broad sampling of genomes, including ancient angiosperm lineages. Recent phylogenetic analyses of the flowering plants have identified Amborella trichopod a, a shrub known only from the island of New Caledonia, as the single sister species to all other living flowering plants. Amborella therefore offers the unique ability to root analyses of all angiosperm features, from gene families to genome structure to morphology, and when coupled with comparisons to non-flowering plants, to reconstruct the features of the 'ancestral angiosperm'. Although the genome structure of the 'ancestral angiosperm' is currently debated, our recent data suggest that a wholegenome duplication predated or coincided with the origin of angiosperms, perhaps catalyzing their success. The
' Amborella Genome Project' is designed to address the fundamental question of genome structure and evolution in the angiosperms, while providing genetic and genomic resources for anyone studying any aspect of angiosperm biology, from morphology to genomics. A better understanding of the structure and evolution of flowering plant genomes will also contribute enormously to society through enhancements to agriculture and forestry.
Patterns of genetic diversity and selection in soybean genome
Lam, H-M 1
SS-M 1
, Xu, X 2 , Liu, X 3 , Song, C 2 , Shao, G 3 , Sun,
1 State Key Laboratory of Agrobiotechnology and School of Life Sciences, The Chinese University of Hong Kong,
2 3 China; BGI-Shenzhen, China; Institute of Crop
Sciences, The Chinese Academic of Agricultural
Sciences, China
Soybean is an important crop for food. Understanding the patterns of genetic diversity and selection in soybean genome will help to locate regions associated to important agronomic traits. To achieve this end, we resequencing 17 wild and 14 cultivated soybean genomes
(~5X depth and >90% coverage for each accession) and generated a total of ~180 Gb sequencing data, using the

Illumina Genome Analyzer II platform. The genomic variations between wild and cultivated soybeans were mapped. The genome-wide re-sequencing data reveal that there is an alarming loss of genetic diversity in cultivated soybeans and that wild germplasms are valuable sources of novel alleles and genes. The extreme high linkage disequilibrium in the soybean genome suggests that marker-assisted breeding will be far less challenging than using map-based cloning with genetic populations. A total of 205,614 single-nucleotide polymorphism tags were identified to facilitate future breeding and quantitative trait analysis. In addition, we also performed de novo sequencing of one wild accession
(~80X coverage) and the genome assembly is now close to completion. We also obtained transcriptome data to enhance the accuracy of gene annotation of the wild genome. By a detailed comparison between the wild soybean genome and the reference cultivated soybean genome using the de novo sequencing and the gene expression data, we can identify expressed genes that are unique to wild or cultivated soybeans. [This work was partly supported by the Hong Kong RGC General
Research Fund 468610 (to H.-M.L.), Hong Kong UGC
AoE Center for Plant & Agricultural Biotechnology
Project AoE-B-07/09, and a special fund from the
Resource Allocation Committee, The Chinese University of Hong Kong (to H.-M.L. and S.S.-M.S.). We also acknowledge funding support from National Natural
Science Foundation of China (30725008), the Chinese
973 program (2007CB815703; 2007CB815705), Chinese
Ministry of Agriculture (948 program), Shenzhen
Municipal Government of China, from Shenzhen Bureau of Science Technology & Information, China
(ZYC200903240077A; CXB200903110066A).]
Sequencing giant genomes: second generation
1 sequencing analysis of
Fritillaria
(Liliaceae)
Kelly, LJ 1, 2
RA 2
, Leitch, AR 2
, Fay, MF
Leitch, IJ 1
1 , Macas, J
, Renny-Byfield, S
3 , Novák, P 3
2 , Nichols,
, Neumann, P
Royal Botanic Gardens Kew, Richmond, UK;
Mary University of London, UK; 3
2
3 ,
Queen
Institute of Plant
Molecular Biology, Èeské Budìjovice, Czech Republic
The genus Fritillaria (Liliaceae) is characterized by an exceptionally large range of genome sizes (c.30,000 to c.
80,000 Mb in the diploid species), and includes some of the largest genomes ever recorded in plants. The range of genome sizes in Fritillaria provides an outstanding opportunity to study the processes involved in genome size evolution. Moreover, the phylogenetic distribution of Fritillaria species with large genomes indicates that two independent shifts towards giant genomes have occurred during the evolution of the genus, allowing comparison of the processes involved in these separate instances of genomic expansion. Studies of plants with small (e.g. Arabidopsis thaliana , c. 157 Mb) and medium
(e.g. Hordeum vulgare , c. 5,500 Mb) sized genomes have revealed that changes in genome size are largely governed by the interplay between the amplification and deletion of repetitive DNA. However, it is not clear whether the same processes control the evolution of giant genomes. Using data from second generation sequencing technologies (e.g. 454 sequencing) we are analysing the composition of the most highly represented sequences within the genomes of multiple Fritillaria species in order to gain insights into the evolutionary dynamics of genomic obesity. We discuss the results of these analyses, and their implications for our wider understanding of how genomes evolve in plants.
Geraniaceae, a natural model system for examining
1 plastid genome evolution
Jansen, R 1
University of Texas at Austin, USA
During the past decade there has been a rapid surge in plastid genome sequencing of flowering plants with over
100 species now publicly available. The sampling of species has increased in two important ways: all major lineages of angiosperms are represented and several highly rearranged genomes have been sequenced. Plastid genome sequences have provided important data for improving our understanding of patterns and rates of genome evolution. Gene and intron content and gene order are highly conserved among the early diverging angiosperms and basal eudicots, but over 60 independent gene and intron losses are present in more derived monocot and eudicots clades. Furthermore, a lineagespecific correlation was identified between rates of nucleotide substitutions and genomic rearrangements.
One of the most remarkable lineages is the geranium family (Geraniaceae). Plastid genome sequencing of 27 species from all five genera in this family have revealed several bizarre phenomena, including include multiple and extreme contractions and expansions of the IR resulting in plastomes with both the smallest (0 bp) and largest (74,571 bp) inverted repeats among all land plants, a remarkable accumulation of dispersed repeats associated with changes in gene order, repeated losses and duplications of genes, and unprecedented increases in rates of nucleotide substitutions, especially in ribosomal protein and RNA polymerase genes. Two unusual changes involve putative gene losses of rpoA in
Pelargonium and all 11 ndh genes in one lineage of
Erodium . The implications of these data for understanding mechanisms of plastid genome evolution will be discussed.
Repetitive DNA and its evolution: genomic strategies
1 to understand the processes and exploit biodiversity
Heslop-Harrison, JS 1 , Schwarzacher, T 1
University of Leicester, UK
Repetitive DNA is comprised of sequence motifs that are repeated hundreds or thousands of times in the plant genome, and includes microsatellites, longer tandem repeats or satellite DNA, DNA transposons and retrotransposons. It shows rapid evolution in copy number, location and sequence, with diverse turnover mechanisms which we will show in the talk. Repetitive sequences often mark the major differences between closely related genomes in, for example, crops and their wild relatives in the same or related species, and hence are critical to study in the context of phylogenetic studies at the DNA level. Because of the repetitive nature, it is hard to analyse by next generation or even wholegenome sequencing methods, leading to the numerous
103

breaks in contigs. In this talk, we will discuss the nature of repetitive DNA evolution, amplification and distribution throughout the genomes and how in situ hybridization, BAC analysis and paired-end strategies can enable repetitive DNA diversity to be analysed.
Using a range of different eudicot and monocot species, with and without reference genomes, we have been able to exploit repetitive DNA to generate markers and examine evolutionary diversification mechanisms in crops, during domestication and that are essential to exploit diversity in plant breeding strategies. Related information, the talk, and references will be available from www.molcyt.com
Organellar genome evolution in parasitic plants – assessing patterns of genome reduction and rate acceleration under relaxed selective pressure
Wicke, S 1,3 , Quandt, D 2 , Müller, K 3 , DePamphilis, C
1
Schneeweiss, GM 1
University of Vienna Rennweg, Austria; 2
4 ,
Rheinische
3
Friedrich-Wilhelms-Universitaet Bonn, Germany;
University of Muenster, Germany;
University, USA
4 Penn State
Among land plants, plastidial and mitochondrial chromosomes show highly divergent patterns in terms of genome structure and substitution rates. While the multipartite plant chondriome exhibits and extraordinary variability regarding size, structure and gene content, the quadripartite structure of the plastid chromosome and its gene content and order is rather conserved. For example, the transition from water to land of green plants is only accompanied by few distinct changes in the plastid genome including at least 8 inversions of plastid chromosomal fractions, some gene transfers to the nuclear genome as well as one gene gain. The present study illustrates the evolution of plant plastomes and chondriomes comparing genome structure and patterns of molecular evolution, with a focus on rate heterogeneity among distinguished gene classes, single genes and gene operons. Reviewing the functions of proteins in the plastid genomes shows that transmembrane proteins as well as proteins required at high copy numbers are retained in the plastid genome. Other genes, however, might be substituted or bypassed by nuclear paralogues which could contribute to the loss of all subunits of an entire protein complex (e.g. genes for ndh-complex, sulfur transporter genes) multiple times during land plant evolution in independent lineages. As a group of plants that exhibit major changes in plastid genome structure, both semi- and non-autotrophic plants will be discussed in order to explore and understand the subtle but continuous reduction of genomes under relaxed evolutionary constraints. The transition from a fully autotrophic way of life towards a complete heterotrophic lifestyle (i.e. complete loss of photosynthesis) via various levels of semi-autotrophy severely affects plastome stability and evolution. To this end, the organellar genomes from several hemiparasitic and holoparasitic representatives from the broomrape family
(Orobanchaceae) have been sequenced using different sequencing strategies. Organellar genomes have been reconstructed de novo and analyzed with respect to colinearity, gene content, pseudogenization and substitution rates. Relaxation of evolutionary constraints appears to
104 occur much earlier during the evolution of parasitism than previously assumed and strongly affects structural integrity of distinct plastome fragments. It seems as if the establishment of obligate parasitism can be viewed as the major prior relaxing the selective pressure on plastome integrity preceding the loss of photosynthesis. The pattern of pseudogenization and rate acceleration is highly lineage specific among different broomrapes.
However, there is a general trend towards the early loss of subunits encoding an eubacterial-type polymerase, and protein subunits involved in electron flow.
Orobanchaceae mitochondrial genes and genomes are apparently not affected by different grades of heterotrophy. They do not exhibit significant alterations in structure and gene content compared to autotrophs, although there is evidence that holoparasites evolve more conservative than hemiparasites. Interestingly, however, is that mitochondrial plastid-derived genes show substitution rates similar to those of plastid genes in holoparasitic lineages.
Evolutionary genomics of diploid
Fragaria
species: insights into the origin of the cultivated strawberry
Liston, A 1 , Govindarajulu, R 2 , Ashman, T-L 2
1 Oregon State University, Corvallis, Oregon, USA;
2 University of Pittsburgh, Pittsburgh, Pennsylvania, USA
The domestication of strawberry ( Fragaria × ananassa ) in the 1700s is well documented, but we have a poor understanding of the evolutionary history of the species that contributed to this important crop. Fragaria
× ananassa originated in Europe from hybridization between two octoploid species, the South American
Fragaria chiloensis and the eastern North American
Fragaria virginiana . Phylogenetic analysis based on plastid genomes indicate that the octoploids are monophyletic, and identify the northwest North
American diploid F. vesca subsp.
bracteata as their cytoplasmic donor. The 240 Mbp genome of the
European diploid Fragaria vesca subsp. vesca has been recently published, and we have used this as a reference to conduct low coverage genome sequencing of F. vesca subsp. bracteata , and two East Asian diploids, F. mandshurica and F. iinumae . Our phylogenomic comparison of 21,563 genes in these four taxa revealed genome-wide patterns of lineage sorting and identified loci that have apparently undergone positive selection.
These results provide novel insights into the genes and genomic regions that were involved in the origin of the octoploid progenitors of the cultivated strawberry.
Unravelling apomixis in aposporous
Hieracium
1
Koltunow, AMG 1 , Johnson, SD
K 3 , Suzuki, G 3
1 , Okada, T 1 , Hu, Y 2 , Ito,
Commonwealth Scientific and Industrial Research
2
Organization Plant Industry, Glen Osmond, Australia;
College of Life Science, Capital Normal University,
Beijing, China; 3 Laboratory of Plant Molecular Genetics

Division of Natural Science, Osaka Kyoiku Universitya,
Osaka, Japan
In apomictic Hieracium subgenus Pilosella species, asexual seed formation is the dominant reproductive mode, however, rare sexual seed are also produced.
Sexual reproduction initiates first in apomicts as the megaspore mother cell (MMC) undergoes meiosis in the ovule. Somatic aposporous initial (AI) cells develop nearby and the sexual pathway is terminated as AI cells undergo mitotic embryo sac formation. Seed formation is fertilization independent. Two dominant, independent loci, LOSS OF APOMEIOSIS (LOA) and LOSS OF
PARTHENOGENESIS (LOP) control apomixis in
Hieracium praealtum . The functions of LOA and LOP and their interaction with the sexual pathway were examined using irradiation mutants, wild-type genetic populations and transgenic analyses. Sexual initiation is required for apomixis initiation because inhibition of
MMC function using a cytotoxic reporter inhibits AI cell differentiation. LOA is sporophytic in action and stimulates both AI formation and sexual suppression.
LOP is gametophytic in action, stimulates fertilizationindependent embryo and endosperm formation and these determinants are tightly linked. Loss of function in either
LOA or LOP results in partial reversion to sexual reproduction. Loss of function in both results in complete reversion to sexual reproduction indicating it is the default reproductive mode. Therefore, LOA and LOP are not essential, mis-regulated elements of the sexual mode of reproduction as some models propose. Sexual and apomictic pathways co-exist, operating in parallel in H. praealtum ovules. Incomplete functional penetrance of these dominant apomixis alleles is likely to give rise to the observed rare sexual progeny. We have identified the genomic region containing the LOA locus and found it to be located on a single chromosome. Transcriptomic analyses are providing information concerning gene expression in AI cells as they transition to aposporous embryo sac formation. This and our progress towards identification of the genes responsible for apomixis initiation will be discussed.
Searching for elements of apomixis in
Zea mays
Brunner, A 1 , Barrell, P 1 , Kessler, S 1 , Grossniklaus, U 1
1 University of Zürich, Zürich, Switzerland
The introduction of apomixis into sexual crop plants bears tremendous promise for agriculture. Most flowering plants reproduce sexually and viable seed formation depends on the fertilization of the egg and central cell, and the coordinated development of the embryo, the endosperm, and the maternal seed coat.
Apomictic plants employ a closely related, yet asexual strategy, in which progeny of maternal origin are produced in the absence of meiotic reduction and fertilization. We proposed that this alternative approach to seed production arose from the spatial and temporal deregulation of ancestral sexual processes (Spillane et al.). Thus, apomixis and sexual reproduction are closely interrelated and tractable, sexual model systems can be used to identify genes that play a role in apomixis. Using maize as a model, we concentrate on two of the three elements of apomixis: apomeiosis and parthenogenesis.
Using transposon-based, forward genetic screens have identified genes that, if mutated, display aspects of diplospory or parthenogenesis. Our progress in the characterization of these maize mutants will be reported.
Targeted sequencing of a chromosome transmitting apomixis
Ozias-Akins, P 1 , Conner, J 1 , Zeng, Y 1
1 The University of Georgia, USA
Apomixis (asexual reproduction through seeds) in
Pennisetum results from aposporous embryo sac development and parthenogenesis of the unreduced egg.
Fertilization of the central cell is required for seed formation. In crosses between the apomictic wild species,
P. squamulatum , and cultivated pearl millet, P. glaucum , a single chromosome is necessary and sufficient to transmit apomixis. From mapping in interspecific F1 hybrids, results indicate that approximately ¼ of the chromosome lacks recombination and is transmitted as a linkage block. Various strategies are being pursued to sequence the chromosome transmitting apomixis including the non-recombining region. These are sequencing of bacterial artificial chromosome (BAC) clones mapping to the non-recombining region, DOP-
PCR products from the microdissected chromosome, and transcriptomes from the apomictic parent and an introgression line containing only the apomixistransmitting chromosome. Candidate genes such as
BABY BOOM have been discovered through sequence analysis. Comparative analyses with sequenced genomes and candidate genes in related species also have been informative for understanding the evolution of this genomic region.
Hybridization delays the sexual development and triggers apomixis in the
Ranunculus auricomus complex
, Pellino, M 2 , Vogel, H 2 , Sharbel, TF 2 ,
1
Hojsgaard, DH 1
Hoerandl, E 1
Dept of Systematic and Evolutionary Botany, Faculty
Center of Biodiversity, University of Vienna, Vienna,
Austria; 2 Apomixis Research Group, Dept of
Cytogenetics and Genome Analysis, Leibniz Institute of
Plant Genetics and Crop Plant Research (IPK),
Gatersleben, Germany
Apomixis, the asexual pathway for seed formation, involves a bypass of meiosis (apomeiosis) and formation of an unreduced female gametophyte. The unreduced egg cell develops parthenogenetically (by pseudogamy or autonomously) into an embryo that is genetically identical to the mother plant. The connection between polyploidy to apomixis is frequently explained in terms of hybridization. We hypothesize that hybridization of different ecotypes and/or polyploidy cause asynchrony of reproductive developmental steps to thus trigger apomixis. In the Ranunculus auricomus complex, the diploid R. carpaticola and tetraploid R. cassubicifolius are the sexual parental species of the natural allohexaploid apomictic nothotaxon R. carpaticola × R. cassubicifolius . To understand and explore the origin and functionality of apomixis we compare the sexual parents with the natural hybrid. Furthermore, we have generated
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laboratory crosses between the early flowering diploid sexual species R. notabilis with the late flowering species
R. carpaticola and R. cassubicifolius . Embryological analyses of the experimental diploid hybrids shows the occurrence of aposporous initial cells as first step in apomictic development, and asynchronous development during megagametogenesis compared with the parental species. The frequency of aposporous cells is lower in the experimental hybrids than in the natural hybrids, and according to flow cytometric seed screening (FCSS), the aposporous pathway is only stable in the natural allohexaploid hybrids since fully-developed apomictic seeds were not recovered in the experimental hybrids.
However, in natural apomictic genotypes, precocious apospory exists and outcompetes sexual development when both pathways are expressed in the same ovule.
Alterations of gene expression and asynchrony in hybrid genomes might be necessary for initiation of apospory, while polyploidy could favour selection of efficient apomictic development. We have thus performed
Illumina transcriptome sequencing in 3 sexual and 2 apomictic accessions to develop a Ranunculus -specific expression microarray. Stage-specific expressionprofiling will inform us to what extent different patterns in gene expression are correlated with asynchronous development to make functional apomixis possible.
Molecular evolution and comparative genomics of soybean orthologs of meiotic genes responsible for gamete reduction
Sherman-Broyles, S 1 , Lee, M-J 1
Doyle, J 1
1 Cornell University, USA
, Mardonovich, S 1 ,
Two recently described mutants of the Arabidopsis genes
OMISSION OF SECOND DIVISION 1 (OSD1) and
PARALLEL SPINDLE 1 (PS1) result in unreduced gametes. We have investigated the evolution of these genes in the duplicated genomes of soybean and its
Australian wild relatives. Cell cycle and meiotic genes have been described as two classes of duplication resistant genes. The genus Glycine has experienced three rounds of whole genome duplication (WGD), the rosiid gamma event (130–240 million years ago (MYA)), the legume event (59mya) and the Glycine event (13 mya).
Arabidopsis has also experienced three WGD events, the rosiid gamma duplication, a beta duplication for which the date is uncertain, and the alpha duplication that occurred 20–100 mya. OSD1 and PS1 show different patterns of retention in Arabidopsis . Arabidopsis retains two homoeologs from the alpha duplication event that show evidence of subfunctionalization, OSD1 is involved in meiosis and UVI4 is involved in endoreduplication.
PS1 is single copy in Arabidopsis but retains two homoeologs in soybean. We report on the sequence diversity, gene phylogenies and expression of these two genes in the genus G lycine r epresented by cultivated soybean, G. sojae , the annual wild relative of soybean and seven species of perennial Glycine .
Marchantia
as a model representing the earliest land plants
Graham, L 1 , Graham, J 2
Cardona-Correa, C 1
, Hanson, D 2 , Cook, M 3 ,
1 University of Wisconsi, USA;
Mexico, USA; 3
2 University of New
Illinois State University, USA
The molecular phylogenetic literature indicates that liverworts form a monophyletic group that is the earliestdiverging clade of modern land plants. The modern paleontological literature provides evidence that liverwort-like plants were present during the Ordovician, a time when their spread could have been fostered by warm, moist conditions that prevailed until the onset of epoch-ending global glaciations. Recent fossil discoveries, as well as structural and stable isotopic comparisons of particular fossil remains to modern
Marchantia suggest that similar liverworts may have been common in the Silurian and Devonian, possibly occupying large areas of terrestrial surface prior to and during the rise of tracheophytes. Liverwort-like early land plants thus early acquired key terrestrial adaptations
– some likely inherited by descendent lineages – that fostered clade persistence through stressful end-
Ordovician glaciations and other periods of dramatic environmental change, to the present time. Identifying such traits is not only useful in modeling the earliest impacts of land plants on global biogeochemistry, but also essential to predicting ways in which current and future environmental change will influence vegetation.
To identify key traits and the order of their appearance, we compared structural, reproductive, and physiological features of Marchantia to those of earlier-diverging liverworts, early-divergent mosses such as Sphagnum , and charophycean green algae grown under subaerial conditions. We have determined that Marchantia and relatives are capable of mixotrophy – the uptake and utilization of exogenous organic compounds such as sugars – indicating that mixotrophy is an early-evolved plant trait that may subsidize the production of increased amounts of tissue that is resistant to microbial attack. By this means, mixotrophy fosters both the ecological persistence of modern Marchantia , and the formation of fossil remains by Marchantia -like ancient bryophytes.
Our recent experimental advances include new evidence that the direction of incident light is the primary environmental signal determining the dorsiventral body orientation of asexual gametophytes produced by
Marchantia and other liverworts, explaining the typical downward growth of rhizoids from lower surfaces and production of gemmae on upper surfaces. A major role for gravitropism in body orientation was disproved by the re-orientation of plant growth in response to light provided only from below, with rhizoids produced from plant surfaces facing away from both light and the gravity vector. Light-directed reorientation allows displaced plants to recover normal spatial orientation, a trait that might have been particularly useful to
Ordovician plants growing on then globally-extensive, but easily disturbed terrestrial quartz sand deposits. Other
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new experimental results derived from comparative analyses illuminate the evolutionary origin and past and present ecological importance of asexual gemmae, bacterial associations, desiccation tolerance, and responses to combined thermal and UV stress, thereby providing examples of ways in which Marchantia models early land plant structure, reproduction, and physiological ecology. Such information can be used to build a conceptual foundation to aid studies focused on the evolution of plant systems biology.
Mutualistic mycorrhiza-like symbiosis in
Marchantia
Beerling, D 1 , Humphreys, C 1 , Bidartondo, M 2
1 University of Sheffield, UK; 2
, Leake, J 1
Royal Botanic Gardens,
Kew, UK
Over thirty five years ago it was hypothesized that mutualistic symbiotic soil fungi assisted land plants in their initial colonization of terrestrial environments. This important idea has become increasingly established with palaeobotanical and molecular investigations dating the interactions between arbuscular mycorrhizal fungi
(AMF) and land plants to at least 400 Ma, but the functioning of analogous partnerships in ‘lower’ land plants remains unknown. Here, we show with multifactorial experiments that colonization of Marchantia , a complex thalloid liverwort, with AMF significantly promotes photosynthetic carbon uptake, growth and asexual reproduction. Plant fitness increased through fungal-enhanced acquisition of phosphorus and nitrogen from soil, with each plant supporting 100-400 m of AMF mycelia. A simulated CO
2
-rich atmosphere, similar to that of the Palaeozoic when land plants originated, significantly amplified the net benefits of AMF and likely selection pressures for establishment of the symbiosis. Our analyses provide essential missing functional evidence supporting AMF symbionts as drivers of plant terrestrialization in early Palaeozoic land ecosystems.
The genome of
Marchantia polymorpha
L.
Yamato, KT 1
Bowman, J 3
, Ishizaki, K 2 , Kohchi, T 2 , Floyd, S 3 ,
1 Kinki University Faculty of Biology-Oriented
2 Technology and Science, Kinokawa, Japan; Kyoto
University Graduate School of Biostudies, Kyoto, Japan;
3 Monash University School of Biological Sciences,
Melbourne, Australia
The liverwort Marchantia polymorpha L. is one of extant species of the first land plants that appeared about 450
Myr ago. The genomes of plastid, mitochondria and Y chromosome in M. polymorpha were the first to be published among all plant species, making significant contributions to plant biology. M. polymorpha has now become a fascinating model organism for plant biology, because of its crucial position in the evolution of land plants and molecular genetic tools that have been recently developed. M. polymorpha is dioecious, and its complete haploid set of chromosomes (approximately
280 Mb) consists of eight autosomes and a single sex chromosome: an X chromosome for a female (n=8+X) and a Y chromosome for a male (n=8+Y). A genetic map has been constructed using >100 markers based on DNA polymorphisms between our two laboratory lines, the number of its linkage group is eight, which is consistent with the number of autosomes. This map can be used for arranging genomic sequences, as well as mapping mutations. Since the Y chromosome has been sequenced, a female standard line of M. polymorpha was selected for whole-genome sequencing at the Joint Genome Institute
(JGI) in USA (PI: J.L. Bowman, Monash University,
Australia) (assembled data not available at the time of abstract submission). To preview the M. polymorpha genome, 30 genomic PAC clones which collectively represent approximately 3 Mb of the genome were sequenced and examined. Based on the obtained data, the
GC content of the M. polymorpha genome is approximately 42%, and the frequency of known transposable elements in the genome was roughly 1 in
100 kb, with an exception of one clone that contains a much larger number of them. Similarity search and EST mapping analyses revealed that the density of proteincoding genes is approximately one per 14 kb. Therefore, the total number of genes in the M. polymorpha genome was estimated to be around 20,000, which is consistent with an estimate from EST analyses. An increasing number of studies indicate the genetic redundancy in the
M. polymorpha genome is low. In angiosperms, many regulatory genes are encoded by gene families, which provides more elaborate regulatory systems. For example, Arabidopsis has five phytochrome genes that were generated by gene duplication and have functionally differentiated. In contrast, M. polymorpha appears to have a single phytochrome gene thus far. Our preliminary data strongly suggest that M. polymorpha has a minimum number of member genes for regulatory systems, such as development and environmental response. It should be also noted that species-specific gene duplication observed in the moss Physcomitrella patens does not appear to be common in M. polymorpha .
Its low genetic redundancy again makes M. polymorpha an excellent model organism. In addition to genomic sequence data, a large number of ESTs have been generated (~300,000 of Sanger reads and >2,000,000 of
454 reads) and will be mapped on genomic data. Since the genome project is ongoing, a more comprehensive set of data is expected to be presented.
Transgenesis of the liverwort
Marchantia polymorpha and its application to developmental genetics
Ishizaki, K 1 , Masuda, A
Yamato,KT 1 , Kohchi, T 1
1 , Saida, Y 1 , Ueda, M 1 ,
1 Graduate School of Biostudies, Kyoto University,
Japan; 2 Graduate School of Science, Kyoto University,
Japan
Transformation technique is an essential component of functional genomics. We have recently developed a simple and rapid Agrobacterium -mediated transformation system for the liverwort Marchantia polymorpha .
Hundreds of stable transformants per sporangium were obtained using immature thalli developed from spores. It took only 3–4 weeks to establish individual transgenic plants after starting liquid culture of spores. DNA analyses verified random integration of 1–5 copies of intact T-DNA between the right and the left borders into the nuclear genome. To avoid potential chimerism, clonal
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transgenics can be rapidly established by using gemmae which arise asexually from single initial cells. Four kinds of selectable markers and various reporters including green fluorescent protein (GFP),
β
-glucuronidase (GUS), and luciferase (LUC), have been successfully used for.
Therefore, it has become feasible to perform various transgenic experiments using M. polymorpha ; for example, overexpression, promoter analysis, and observation of subcellular protein localization.
Agrobacterium -mediated transformation is a powerful tool not only for introduction of constructs of interest into plants, but also for T-DNA insertional mutagenesis.
We have generated over 10,000 T-DNA tagged lines of
M. polymorpha using the binary vector pCAMBIA1300, and screened them for abnormal morphological phenotypes. Because of the haploidy of M. polymorpha , their mutant phenotypes were detectable in the primary generation of transformants. We have isolated 22 lines that exhibited various morphological phenotypes, such as formation of callus-like tissue, loss of air-chamber development, and defect in gemma formation.
Characterization of these mutants is expected to reveal both the common and divergent developmental programs between gametophyte-dominant and sporophytedominant land plants. We will present our recent progress in the genetic characterization of these M. polymorpha morphological mutants.
The role of patterning genes in
Marchantia
: implications for body plan evolution in land plants
Floyd, S 1 , Bowman, J 1
1 Monash University, Vic, Australia
The availability of genetic and genomic resources and tools for Marchantia polymorpha allow the analysis of gene function in a representative of the earliest-diverging land plant clade. Interpretation of mutant and transgenic phenotypes and expression data indicate that in
Arabidopsis the Class III Homeodomain-leucine zipper
(Class III HD-Zip) family of transcription factors are essential for SAM formation, patterning of vascular tissues, adaxial/abaxial polarity in leaves and cotyledons, and lateral root formation. With a role in most major organs of the sporophyte, it is likely that this family of transcription factors has been important in land plant evolution. A survey of these genes in land plants suggested that a role in vascular patterning and meristem function is conserved in vascular plants but so far functional data are lacking outside of flowering plants. I will present data describing the role of the single Class
III HD-Zip gene in Marchantia . Our findings have implications not only for the ancestral function for the
Class III Hd-Zip, but the significance of miRNA regulation in embryophyte development and evolution.
1
Developmental gene regulation by chromatin remodelling factors in basal land plant
Marchantia polymorph a
Dierschke, T 1 , Bowman, J 1
School of Biological Sciences, Monash University, Vic,
Australia
Chromatin remodelling factors are master regulators of gene expression in animals as well as in plants. Research in Arabidopsis thaliana and Physcomitrella patens indicate that these genes are responsible for major transitions in developmental stages during the life cycle.
Arabidopsis thaliana CURLY LEAF (CLF), a homologue of Drosophila ENHANCER OF ZESTE
(E(z)) and member of the Polycomb Repressive Complex
2 (PRC2), is necessary for stable repression of floral homeotic genes. In Drosophila , TRITHORAX Group proteins (TrxG) often oppose the action of Polycomb group (PcG) proteins. Mutations in ATX1 (a TrxG gene) suppress phenotypic defects of clf mutants, providing evidence that PcG/TrxG factors have antagonistic functions in plants by regulation of a shared gene locus.
Each of the three Arabidopsis E(z) homologues CURLY
LEAF (CLF), MEDEA (MEA) and SWINGER (SWN) displays different spatio-temporal expression pattern during plant development and all PRC2-like complexes display variable compositions of the core proteins, demonstrating that CLF, SWN and MEA act partially redundant in A. thaliana . In P. patens the PcG proteins
FIE and PpCLF, both proteins being components of
PRC2 in Arabidopsis , play a key role in phase transitions in the life cycle of P. patens , including the alternation of generations. The basal land plant Marchantia polymorpha is a good model as EST´s are available and its genome has recently been sequenced. Unlike A. thaliana , M. polymorpha shows no redundancy in PRC2 proteins, providing us with the opportunity to study the ancestral role of these genes in land plants. We are undertaking a combination of gene expression analysis and phenotypic analyses of gain-and induced loss-offunction alleles that will provide insights in the roles of the chromatin remodelling factors MpE(z), MpTRX and
MpGYM in the life cycle of M. polymorpha .
Decoding plant environmental strategies with new models for the 21st century
Cheeseman, J 1 , Dassanayak, M 1 , Oh, D-H 1 , Bohnert, H 1
1 University of Illinois, USA
There is no doubt that for the last 20 years, a single major model plant system such as Arabidopsis was well justified: the technical limitations of the early molecular era and first generation sequencing made it impossible to maintain the taxonomic breadth previously characterizing the field of plant physiology. Now, however, the key obstacle to progress in understanding genetic mechanisms by which plants tolerate and ameliorate stress is the fact that all of the current major models are plants adapted to and dependent on abundant resources.
Plants adapted to extreme conditions – those which actually have successful environmental response strategies – have been significantly under-utilized. It is, therefore, opportune at this point to identify those characters to be sought in defining the next generation of plants for which serious genetic, proteomic, metabolomic and physiological resources should be developed. This
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contribution will address four major areas, including those directly relate to stress tolerance, to genetic tractability, to amenability to systems biology approaches involving multiple –omics, and to those related to life history strategies and life styles. The need for multispecies breadth in the post arabidocentric universe will be presented.
Cluster root molecular physiology: regulating the development of an adaptation that mines phosphorus from nutrient-impoverished soils
Finnegan, PM
Lambers, H 1
1 , Jost, R 1 , Siddique, ABM 1 , Ludwig, M 2 ,
1 School of Plant Biology, University of Western
2 Australia, Australia; School of Biomedical,
Biomolecular and Chemical Sciences, University of
Western Australia, Australia
Ancient, highly weathered soils, such as those found in south-western Australia and the Cape region of South
Africa, contain relatively low phosphorus and are extremely poor in plant-available inorganic phosphate
(Pi). Evolution has provided plants endemic to these regions with structural and physiological adaptations to maximise their capacity for acquiring Pi. One of these adaptations is the cluster roots found among Proteaceae.
Cluster roots typically exude substantial quantities of carboxylates that mobilise complexed or sorbed Pi into the soil solution. Once liberated from soil particles, the Pi must be taken across the plasmalemma of cells in contact with the soil solution and redistributed throughout the plant body. We are exploring the molecular physiology of Pi acquisition by 'proteoid' cluster roots and the underlying molecular genetic control of their development in Hakea prostrata . Pi transport across membranes is most likely through members of the PHT1 family of Pi transporters. In those species examined, the steady-state transcript abundance of several members of the PHT1 gene family, especially those expressed in roots, respond to Pi in the soil solution such that transcript accumulation is repressed by high Pi availability. It is unknown if this down-regulation of Pi transporters occurs in H. prostrata and is of interest because previous work showed that this plant, unlike most other plants investigated, seems unable to effectively down-regulate Pi up-take in response to excess Pi (M. W. Shane, M. E. McCully, H. Lambers
(2004). This situation can lead to Pi toxicity through the accumulation of excess Pi in leaves, if Pi availability is raised even slightly above the Pi levels found in this species' natural habitat. The PHT1 gene family of H. prostrata contains at least 13 members. Complementary
DNAs for four of the 13 HpPHT1 genes were isolated from cDNA libraries derived from cluster roots, indicating that these genes are expressed in this tissue.
We are continuing to characterise the PHT1 gene family of H. prostrata to determine the tissue-dependent transcript profiles and Pi responsiveness of its members.
The Pi responsiveness of PHT1 genes in model plants relies on a complex regulatory network that involves proteins and microRNAs. We are undertaking a transcriptome sequencing approach to determine whether the genetic functions known to be involved in regulating
Pi homeostasis in other model species are present in H. prostrata . This analysis will give insights into the evolution of Pi responsiveness in the Proteaceae, and may give insights into why H. prostrata is unable to regulate its Pi responsiveness as effectively as other plants, while at the same time displaying a very effective
Pi acquisition capability.
Evolution of salt tolerance: a phyloinformatic approach to understanding the evolution of complex traits to adapt to harsh environments
Bromham, L 1 , Bennett, T 1 , Flowers, T 2
2
1 Australian National University, Canberra, Australia;
University of Sussex, UK
Salt-affected land is a serious and increasing problem in agriculture and land management, but the large amount of effort directed at developing salt-tolerant crops has produced disappointingly few successes. This is perplexing, given that there are over 1500 plant species that have some natural salt-tolerance. To resolve this paradox, we need to understand why so many different lineages have produced naturally salt-tolerant species, yet why we have been limited in our ability to repeat this success in artificial breeding and associated genetic techniques. We take a comparative, biodiversity-based approach to understanding the evolution of salt-tolerance, surveying all halophytic flowering plant species and asking what traits or preconditions underlie the evolution of salt-tolerance.
Using
Arabidopsis
gene networks to study
1 transcriptome dynamics and stress responses
Ma, S 1 , Bohnert, H 2 , Dinesh-Kumar, SP 1 , Oh, D-H
University of California at Davis, USA; 2
1
University of
Illinois at Urbana–Champaign, USA
Arabidopsis gene networks based on large scale gene expression data have been constructed with various methods, such as relevance network and graphical
Gaussian model. However, little is known about the molecular mechanisms behind these networks. One hypothesis is that the large network can be separated into many coherent sub-networks whose genes’ expressions are driven by various hub proteins, especially hub transcription factors (TFs). To identify such hub TFs, we developed an algorithm to identify promoter motifs associated with the genes in the network, and identify the motifs that would bind to the hub TFs, which we call them hub motifs. Once the motifs are identified, a protein array based method was used to identify the hub TFs that bind to these hub motifs. Such methods are helpful to elucidate the transcriptional modules Arabidopsis uses to counteract various abiotic and biotic stresses.
The genome of an extremophile,
Thellungiella parvula
(Brassicaceae)
Dassanayake, M 1 , Oh, D-H 2
, Bohnert, H 2
, Haas, J 1 , Hernandez, A 1 ,
1
Cheeseman, J 1
University of Illinois, Urbana–Champaign, Illinois,
USA; 2 4Gyeongsang National University and WCU
Program, Government of Korea, KR
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A close relative of Arabidopsis thaliana , Thellungiella parvula is adapted to an extremophile lifestyle. T. parvula is, is notable for its tolerance of high salinity, freezing temperatures, water deficit and resource-poor environments. With a small nuclear genome of ~140Mb in 7 chromosomes (N), T. parvula represents an extremophile model species. We have determined its genome sequence and the transcript profile with the objective of comparing how the plant differs in genome structure and expression characteristics from the model
Arabidopsis , which is a glycophyte. Arabidopsis and
Thellungiella ancestors are estimated to have separated
~15–20 MY ago. A comparison of their respective genomes provides information that will allow for an analysis of the evolutionary pressures that shaped the two species leading to divergent lifestyles. Our sequencing strategy was aimed to retrieve 50x coverage of the estimated genome size by a hybrid sequencing approach utilizing only next generation sequencing technologies:
454 Roche GS FLX Titanium and Illumina GA2. Our de novo assembly of short read sequences provided a high quality draft genome of T. parvula that shows extensive microsynteny with Arabidopsis and Brassica chromosomes. In many instances the observed degenerate synteny with Arabidopsis is due to the absence of close homologs of T. parvula in Arabidopsis .
However, over 70% Arabidopsis transcript models have inferred homologs sharing more than 75% sequence similarity with T. parvula transcripts. Functional categories that are significantly differently represented in
T. parvula compared to Arabidopsis include genes for transporters and receptor signaling functions. Examples of genome structure, gene complement and gene expression, and some obvious differences between T. parvula and A. thaliana , will be further discussed. Our results highlight differences between an extremphile and glycophyte lifestyles at two key levels: first, copy number variation in terms of gene copy number which affect expression strength and second, allelic variation that translates to changes in protein domain structure, where stress-essential proteins show minute alterations from their glycophytic homologs. On the level of future crop breeding in the age of genomics, we argue for the importance of selecting extremophile characters in crop species that could even emerge from wild but stressadapted species.
Auxin response and signaling factors in
Marchantia polymorph a
Kato, H 1 , Ishizaki, K 1
Kohchi, T 1
, Nonomura, M 1 , Yamato, KT 1 ,
1 Graduate School of Biostudies, Kyoto University, Japan
Phytohormone auxin regulates a variety of growth and developmental processes in plants. Recent studies in
Arabidopsis have provided a scheme that F-box proteins,
TIR1/AFBs, function as auxin receptors and degrade transcriptional repressors, AUX/IAAs, allowing ARFs to regulate transcription of auxin-responsive genes. The liverwort Marchantia polymorpha , which belongs to a
110 group of basal land plants, has low genetic redundancy and undergoes simple developmental processes. We have been investigating the mechanism of auxin signaling in
M. polymorpha to understand if the components of the auxin signaling mechanism are conserved among land plants. Application of synthetic auxin, 1naphthaleneacetic acid (NAA), induced development of rhizoids from not only the ventral but also the dorsal side of thallus at 1
µ
M. In addition, treatment with auxin transport inhibitor, 2,3,5-triiodobenzoic acid (TIBA), occasionally caused inversion of the dorsiventrality in thallus. These results suggest that auxin gradient is involved in formation of the dorsiventral axis in M. polymorpha . Next, we examined the distribution of auxin in M. polymorpha by monitoring an auxin-inducible reporter gene fusion, GH3:GUS. Transgenic plants carrying GH3:GUS showed elevated GUS activity in response specifically to auxin treatment, indicating the
GH3:GUS reporter system can measure auxin distribution in M. polymorpha . Histological GUSstaining was observed at the bottom of gemma cups and in developing sporophytes without exogenous auxin, suggesting a role of auxin in gemma and sporophyte development. To investigate the molecular mechanism of the auxin signaling in M. polymorpha , we searched for key genes in auxin signaling. We identified one
AUX/IAA (MpIAA), three ARFs (MpARF1, MpARF2, and MpARF3) and one TIR1/AFB (MpTIR1) genes in the M. polymorpha genome. MpARF1 has a putative target sequence for microRNA167 as in ARF6 and ARF8 of Arabidopsis . MpIAA has a glutamine-rich region in addition to the four known functional domains conserved among AUX/IAAs. In Arabidopsis , it is known that mutation in domain II of AUX/IAA confers dominant auxin-resistant phenotype. The expression of MpIAA that has the mutation in domain II conferred reduced sensitivity to auxin. Our results suggest that the molecular mechanism of auxin signaling is conserved among land plants through the evolutionary shift from gametophyte-dominant to sporophyte-dominant life cycle
Auxin signaling factors in
Marchantia polymorpha
II
1
Flores-Sandoval, E
Monash University, Australia
Auxin is involved in establishing key developmental processes in flowering plants but the role of this hormone is not well understood in the earliest diverging lineages of land plants. We test the morphogenetic effect of lossof-function Auxin Response Factors of Marchantia polymorpha by experiments using the TOPLESS corepressor fused to protein-protein interaction domains of
AUX/IAA and ARF proteins. Dominant negative mutations of TOPLESS were also performed to asses gain-of-function ARF phenotypes. The function of individual ARF lineages was assessed in the context of conserved miRNAs 160 and 390 in liverworts. Our work contributes in understanding land plant evolution by comparing morphogenetic effects of auxin signaling disruption between Arabidopsis and Marchantia , which has a haploid dominant thallus with potentially several ancestral characteristics.

Molecular conservation of MIKC*-type MADS-box proteins in the gametophytic generation of land plants: insights gained from studying bryophytes
1
Zobell, O 1,2 , Faigl, W 2 , Muenster, T 2 , Zachgo, S 1
University of Osnabrueck, Osnabrueck, Germany; 2 Max
Planck Institute for Plant Breeding Research, Cologne,
Germany
Land plants are characterized by an alternation of two generations, the haploid gametophyte and the diploid sporophyte. The development of the small and simple male gametophyte of the flowering plant Arabidopsis thaliana critically depends on the action of five MADSbox proteins of the so-called MIKC* subclade. These
MADS-box genes were isolated from land plants with relatively large and complex gametophyte bodies, namely the bryophytes. We found that the gene family expanded in the mosses Sphagnum subsecundum,
Physcomitrella patens , and Funaria hygrometrica , just as it did in any vascular plant analyzed till date. However, only a single homologue, Marchantia polymorpha
MADS-box gene 1 (MpMADS1), has been retained in the liverwort M. polymorpha . Liverworts are the earliest diverging land plants, and so a comparison of
MpMADS1 with its angiosperm homologues addresses the molecular evolution of a land plant-specific transcription factor over the widest phylogenetic distance. MpMADS1 was found to form a homodimeric
DNA-binding complex, which is in contrast to the
Arabidopsis proteins that are functional only as heterodimeric complexes. The M. polymorpha homodimer, nevertheless, recognizes the same DNA sequences as its angiosperm counterparts and can functionally replace endogenous MIKC* complexes to a significant extent when heterologously expressed in
Arabidopsis pollen. The 11 MIKC* homologues from the moss F. hygrometrica are highly and almost exclusively expressed in the gametophytic generation. Taken together, these findings suggest that MIKC* MADS-box proteins have largely preserved molecular roles in the gametophytic generation of land plants. Currently, we are focusing on elucidating the specific role of MpMADS1 in M. polymorpha .
The role of CLE in
Marchantia
development
Sawa, S 1 , Tabata, R 1
1 Kumamoto University, Japan
CLV3/ESR related (CLE) is supposed to function as peptide hormone to regulate various developmental steps in plants. Marchantia has only one CLE gene, MpCLE1.
In order to examine their developmental function, we produced over expression and knock down transgenic
Marchantia plants. MpCLE1 OX plants fail to develop epidermal cell. Air pore like structure was produced.
However, inside of air pore structure can be observed directly from out side. Interestingly, air pore like structure was produced both of abaxial and adaxial side in the MpCLE1 OX plants. In the case of knock down transgenic plants, excess cell division of epidermal cells was observed. Here we introduce our opinion about conservation and difference of CLE signaling molecular mechanisms and functions during plant evolution.
Functional analysis of
MpLFY
, the homolog of
LEAFY
in
Marchantia polymorpha
1
Sakai, Y 1 , Araki, T 1
Dept of Responses to Environmental Signals and
Stresses, Division of Integrated Life Science, Graduate
School of Biostudies, Kyoto University, Japan
LEAFY (LFY) encodes a plant-specific transcription factor that plays a key role in the formation of flowers in angiosperms. LFY orthologs have been found also in gymnosperms, ferns, fern allies, mosses, and liverworts.
In most cases, LFY is a single-copy gene. In Arabidopsis thaliana , transcriptional networks downstream and upstream of LFY are well studied. Floral homeotic genes encoding MADS-box transcription factors, such as AG ,
AP3 , PI , AP1 and CAL , have been identified as direct targets of LFY . On the other hand, several transcription factors including AGL24 , SOC1 and SPL3 are involved in regulation of LFY expression. Recently, function of the LFY orthologs from non-flowering plants was analyzed in detail in Physcomitrella patens . It was demonstrated that Physcomitrella LFY s ( PpLFY1 and
PpLFY2 ) are involved in the cell division of the sporophyte. Although PpLFY s are expressed not only in the sporophyte (diploid) but also in gametophyte
(haploid), their functions in gametophyte are unknown.
We isolated the LFY ortholog, MpLFY , from Marchantia polymorpha , expecting that the study of function of
MpLFY should provide important information to understand the origin, ancestral functions, and evolution of LFY . Where and how does MpLFY function in M. polymorpha ? Whether does LFY function in gametophyte? Furthermore, which kinds of genes are directly regulated by MpLFY ? To answer these questions, we have performed the expression analysis of MpLFY in various tissues throughout the life cycle. Transgenic plants, in which the expression levels of MpLFY are altered, were produced and analyzed. To better understand the phenotypes of these transgenic plants, developmental processes of wild-type plants were observed in organ-, tissue- and cellular-levels, using live imaging. In addition, by microarray expression analysis, using the EST-based custom-made array and a glucocorticoid receptor (GR)-fusion protein system, it was shown that hundreds of genes were up-regulated in haploid gametophyte by activation of MpLFY-GR protein. To identify the direct targets among these candidate genes, their induction plofiles in the transgenic plants expressing MpLFY-GR are investigated in the presence or absence of cychloheximide. Results of these studies will be discussed.
Diversification of post-Golgi trafficking pathways among land plants
Ueda, T 1
1 The University of Tokyo, Japan
Membrane trafficking plays integral parts in various cell activities in all eukaryotic cells. RAB GTPase and
SNARE are evolutionary conserved regulatory molecules for membrane trafficking. In general, RAB GTPase promotes tethering of transport vesicles/organelles to
111

target membranes, which is followed by membrane fusion executed by assembly of specific combinations of three Q-SNAREs (Qa-, Qb-, and Qc-SNAREs) and one
R-SNARE into a stable complex. Recent comparative genomics suggests that the diversification of membrane trafficking pathways was, at least partly, achieved by paralogous gene expansion of organelle identity molecules such as the RABs and SNAREs. However, it remains completely unknown how newly acquired molecules pioneered a new trafficking route. Plants have acquired several endocytic Rab and SNARE molecules unique to plants during evolution, which strongly suggests that endocytic and/or post-Golgi trafficking pathways have been diversified in a plant-unique way.
ARA6 is a plant-unique RAB5-like GTPase of
Arabidopsis thaliana with several unique structural features. Recently, we have found that ARA6 regulates a novel trafficking pathway from endosomes, which also involves a plant-unique longin-type R-SNARE with a characteristic insertion in its N-terminal longin domain.
Plants also harbor orthologs of animal RAB5, which regulate traffic to the vacuole via endosomes. To understand when and how plants acquired the unique endosomal trafficking system, we are studying structures and functions of endosomal RAB and SNARE molecules in the basal land plant Marchantia polymorpha . We isolated homologs of ARA6 and conventional RAB5 in
M. polymorpha , and compared localization and function of their products. As we expected, these two RAB5 groups act in different trafficking pathways in M. polymorpha and, intriguingly, functions of ARA6-type
RAB5s seem to be diversified between A. thaliana and
M. polymorpha .
Developmental and environmental control of CAM expression in Bromeliads: the role of phytohormones and other signaling molecules
Freschi, L 1 , Rodrigues, MA
Hamachi, L 1 , López, AM 1
1 , Mioto, PT 1 , Pereira, PN 1
, Mercier, H 1
1 University of São Paulo, Brazil
,
The extent to which the biochemical and physiological traits of CAM are expressed can greatly vary both within and between species. The so-called C
3
–CAM facultative plants are probably the best examples of intraspecific plasticity in the expression of CAM while, in contrast, constitutive CAM plants are believed to exhibit a more limited flexibility in the proportion of CO
2
uptake over the day and night. Regardless of these differences in classification, the degree of CAM expression in both facultative and constitutive CAM species has been shown to be strongly dependent on the photosynthetic maturity of the tissues. In this study, we have demonstrated that even pineapple ( Ananas comosus ), an archetypical CAM constitutive bromeliad, displays a remarkable flexibility in the expression of CAM during the initial phases of development. In fact, when challenged by a reduction in the water availability, young individuals of this bromeliad are able to respond with an intense, rapid, and
112 completely reversible up-regulation in nocturnal malate accumulation and in the expression and activity of important CAM enzymes, indicating that the expression of CAM represents an adjustable response rather than a predetermined state during the early ontogeny of this species. Similarly, Vrisea gigantea and Guzmania monostachia , two epiphytic tank bromeliads, also showed a high degree of flexibility in CAM expression, which was strongly dependent on both plant and leaf age.
In these species, most of the up-regulation of CAM under water deficit conditions occurred specifically in the upper leaf portion, which is the more mature and light-exposed region of tank-forming leaves. Taking advantage of the photosynthetic plasticity exhibited by these three bromeliad species, we have characterized the hormonal signals associated not only with the intensification of
CAM but also with the attenuation of this photosynthetic pathway in response to changes in environmental factors such as water, light and nutrient availability. This approach allowed us to identify a substantial correspondence between the signals involved in both up- and down-regulation of the CAM expression, characterizing ABA and cytokinins as the main hormonal components of the signal transduction pathways controlling the levels of CAM activity. For instance, during both up- and down-regulation of CAM in pineapple, the degree of CAM expression was positively and negatively correlated with the endogenous levels of abscisic acid (ABA) and cytokinins, respectively.
Moreover, when exogenously applied, ABA stimulated and cytokinins repressed the expression of CAM.
However, inhibition of water-deficit-induced ABA accumulation did not block the up-regulation of CAM, suggesting that a parallel, non-ABA-dependent signaling route was also operating. The involvement of both nitric oxide (NO) and cytosolic calcium during CAM signaling was also demonstrated, revealing that changes in cytosolic calcium represent a convergence point among water stress, ABA and NO during the signaling cascade controlling CAM expression. Finally, the signaling events modulating the degree of CAM expression during the ontogeny of these bromeliad species will be also discussed.
Lending colour to the CAM story…
Ceusters, J
Calcoen, J 1
1 , Borland, AM
, De Proft, MP 1
2 , Taybi, T 2 , Godts, C 1 ,
1 KULeuven, Heverlee, Belgium; 2 Newcastle University,
Newcastle Upon Tyne, UK
The correct functioning of Crassulacean acid metabolism (CAM) hinges on effective temporal coordination of the metabolic processes that determine the supply and demand for carbon over the day/night
CAM cycle. In general, light is considered a crucial input factor in sustaining the circadian rhythms of CO
2
uptake and release in CAM plants, probably by triggering different light sensitive sensors such as the red/far red absorbing phytochromes and the blue/UV-A absorbing cryptochromes. Moreover, light is considered essential for the daytime processing of nocturnally accumulated malic acid, as plants in continuous darkness maintain high levels of malic acid for several days. However, the interplay of the different components of the light spectrum (i.e. different wavelengths of light) with the key

components of CAM, i.e. stomatal aperture, gas exchange, malic acid processing and carbohydrate partitioning is largely unknown. A 24 h experiment was conducted with the CAM plant Aechmea ‘Maya’ under an 18 h photoperiod but with 4 different wavelengths illuminated by LED, i.e. 470 (blue), 530 (green), 630
(orange – red) and 660 (red) nm administered at 10 µmol m -2 s -1 . Control treatments consisted of an 18 h photoperiod under white light (100 µmol m -2 s -1 ) and continuous dark for 24 h. For each regime, gas exchange was monitored by IRGA and leaf samples were taken every 4 hours for the determination of organic acids, sugars and starch. The data was complemented with activity measurements in vitro of the key CAM enzymes
PEPC and PEPCK, along with measurements of protein and transcript abundance of both enzymes over the diel cycle. By integrating measurements of the key components of CAM at the molecular, biochemical and physiological levels, a picture emerges of the complex interplay between light quality and the functioning of this photosynthetic specialization.
Integrated functional genomics of crassulacean acid metabolism in the Common Ice Plant
(
Mesembryanthemum crystallinum
L.)
Cushman, JC 1 , Albion, RL 1 , Gulle, B 1 , Covington, M 2
Schlauch, KA 1
,
1 1Dept of Biochemistry and Molecular Biology, MS200,
University of Nevada, Reno, USA; 2 2Dept of
Biochemistry and Cell Biology, Rice University,
Houston, USA
Crassulacean acid metabolism (CAM) is a specialized mode of photosynthesis that improves atmospheric CO
2 assimilation in water-limited terrestrial and epiphytic habitats and in CO taking up CO
2
2
-limited aquatic environments by
from the atmosphere partially or predominantly at night. Temporal separation of nocturnal
CO
2
uptake and fixation by phosphoenolpyruvate carboxylase (PEPC) and daytime formation of C
4
organic acids (i.e., malic acid), reciprocal fluxes in storage carbohydrates, and inverse stomatal behavior is predicted to be under circadian clock control, however, the molecular genetic mechanisms that underlie the control of these processes remain largely unknown. In order to investigate circadian clock control of CAM, we have initiated an integrated functional genomics program to simultaneously investigate transcriptomic, proteomic, and metabolomic profiling along with metabolic pathway reconstruction to integrate these diverse data sets in the stress-inducible CAM species Mesembryanthemum crystallinum. To date, we have used a combination of
Sanger sequencing and Roche/454 Life Sciences pyrosequencing to generate over 2.3 million EST reads resulting in 765 MB of cDNA sequence data. These data have been used to generate microarray mRNA expression profiles or to serve as a reference transcriptome for high throughput RNA sequencing to characterize circadian clock control of mRNA transcript changes over a 48 or
72-hour period with sampling every 4 hours (under constant light and temperature conditions) in wild type plants performing either C
3
photosynthesis or CAM. A parallel study was also performed with a CAM-defective mutant in order to discern CAM-specific changes from those adaptive responses arising directly from either salinity or drought stress. In addition to mRNA expression studies, we are performing concomitant protein and metabolite profiling studies to identify potential regulatory hierarchies and interactions that might mediate posttranscriptional and posttranslational modes of circadian clock regulation.
Using RNA-seq to dissect the molecular basis for circadian optimisation of crassulacean acid metabolism
Boxall, SF 1
N 1
, Dever, LV
, Hartwell, J 1
1 , Gregory, R 1 , Knerova, J 1 , Hall,
1 Dept of Plant Sciences, Institute of Integrative Biology,
University of Liverpool, Liverpool, UK
The goal of our research is to identify all the genes involved in the establishment and circadian coordination of crassulacean acid metabolism (CAM) in the obligate
CAM species, Kalanchoë fedtschenkoi . K. fedtschenkoi is an excellent model for the study of CAM photosynthesis for a number of key reasons. Firstly, extensive biochemical and physiological characterization of CAM has previously been performed with this species.
Secondly, we have developed a simple, rapid and efficient stable transformation system. Thirdly, it is readily amenable to molecular, biochemical and physiological analysis permitting detailed phenotypic characterization of transgenic lines. Finally, it performs a clear developmental progression from C
3
to CAM during leaf ontogeny. By performing RNA-seq experiments using both the Roche 454 and Applied
BiosystemsSOLiD second generation sequencing systems, we have compared the transcriptome of C
3
and
CAM leaves from K. fedtschenkoi over a diurnal cycle.
Detailed bioinformatic analysis of the RNA-seq data has permitted us to identify thousands of known and unknown genes with potential roles in either the biochemical steps or the circadian coordination and optimization of CAM. We are focusing downstream analysis on genes encoding regulatory proteins that could play a role in the circadian coordination of CAM; undertaking detailed RT-PCR studies to corroborate the
RNA-seq data, and then manipulating putative circadianregulated CAM genes in transgenic lines using both silencing with RNAi, and constitutive over-expression.
In addition, we have also silenced and over-expressed many of the known major metabolic steps of the CAM pathway (PEPc, PPDK, NAD(P)-ME etc), and have begun detailed phenotypic analysis of these transgenic lines. Some of our preliminary phenotypic analysis of the transgenic lines will be presented, along with an overview of the discoveries achieved through the RNAseq analysis of the diurnal and circadian control of the
CAM transcriptome.
Molecular basis of crassulacean acid metabolism
(CAM) evolution inferred from gene family structures in Neotropical orchids (Subtribe Oncidiinae)
Gulle, B 1 , Albion, BL 1 , Silvera, K 2 , Cushman, JC 1
1 Dept of Biochemistry and Molecular Biology, University of Nevada, Reno, NV, USA; 2 Dept of Botany & Plant
Sciences, University of California, Riverside, CA, USA
113

Crassulacean acid metabolism (CAM) is an important water-conserving photosynthetic pathway present in approximately 7% of vascular plant species from at least
35 families. The Orchidaceae is arguably the largest family of vascular plants with about 30,000 species of which approximately 40% are likely to exhibit some degree of weak or strong CAM. Phylogenetic analysis from 1,103 species native to Panama and Costa Rica revealed that CAM has evolved independently several times within the Orchidaceae. The Oncidiinae, which is the second largest orchid subtribe, was selected for evolutionary analysis, because it has one of the most well established molecular phylogenies available and has been sampled by leaf tissue carbon isotope composition. To better understand the CAM evolution, molecular changes were observed using microarray analysis. Roche/454 pyrosequencing for the strong CAM orchid species
Rossioglossum ampliatum resulted in a total of 189 Mb of data with more than 41,000 contigs and 100,000 singletons. More than 39,000 probe sets were used to design Nimblegen oligonucleotide microarray to detect the expression differences among three closely related orchid species performing photosynthesis in a range of
C
3
to weak CAM to strong CAM. In order to test the hypothesis that CAM evolution is accompanied by gene duplication events and recruitment of discrete isogenes with enhanced mRNA expression, we have investigated gene family structures for phosphoenolpyruvate carboxylase (PEPC), carbonic anhydrase (CAH) and glucose-6-phosphate/Pi translocator (GPT). We have sampled over 2000 cDNA products using a degenerate
RT-PCR strategy to recover an average of five PEPC, four CAH and two GPT isogenes within each family across ten orchid species. Ana24-hour gas exchange analysis showed that weak CAM species exhibit limited amounts of nocturnal CO
2
uptake when compared to strong CAM species. Our current results indicate that the evolutionary progression from the ancestral C
3
state to the derived CAM state is accompanied by selective recruitment of discrete isogenes that show increased mRNA expression in order to fulfill the enhanced metabolic demands of CAM to perform nocturnal CO
2 fixation.
Functional genomics of CAM in the monocot biofuel feedstock crop
Agave sisalana
Boxall, SF 1 , Waller, JL 1 , Gregory, R 1
Hartwell, J 1
, Hall, N 1 ,
1 Dept of Plant Sciences, Institute of Integrative Biology,
University of Liverpool, Liverpool, UK
Climate change predictions based on a variety of models suggest that the land area covered by arid and semi-arid regions will expand this century as the climate warms and extreme weather events become more frequent.
Furthermore, there is a growing realisation that the human population will grow to 9 billion by 2050 and that the yield of the world's major food crops has reached a plateau. There are thus grave concerns about the ability of the world's farmers to generate sufficient food to sustain the predicted population. At the same time, concerns that global warming is being driven by CO
2 released by the burning of fossil fuels are driving a push to develop sustainable, carbon-neutral liquid biofuels from crop biomass. Current biofuels largely rely on food
114 crop species such as sugar cane and maize, and thus compete with food security for available land. With all of these major mounting pressures, there is a growing acceptance that 21st century agriculture needs to think outside the box in order to respond to the challenges and balance food security with the production of crop feedstocks for biofuels. Crassulacean acid metabolism
(CAM) is a metabolic adaptation of photosynthesis that increases water use efficiency relative to C
3 photosynthesis by between 6 and 20-fold. Thus, CAM plants are able to thrive on arid and semi-arid land where major crops such as rice and wheat will not grow productively. Certain CAM crops can achieve very respectable yields of up to ~ 45 tonnes per hectare per year on seasonally-dry land. In particular, high productivity species of the monocot CAM genus Agave , such as Agave tequilana, Agave fourcroydes , and Agave sisalana , have been proposed as excellent biofuels feedstocks crops suited to seasonally-dry lands. A key reason why Agaves are believed to have great potential is their high content of the fructan sugars which can very readily be fermented into ethanol. In addition, they have a high percentage of low lignin cellulose (e.g. sisal fibre), which could be exploited with second-generation biofuels technology aimed at the conversion of cellulose to ethanol. Despite the urgent and pressing need to optimize the exploitation of certain CAM plants for human good, they remain one of the most poorly understood photosynthetic types and little scientific research or breeding has been done to enhance the productivity of CAM crops. Our goal is to develop and exploit genomic resources for CAM species. In particular, we are using second-generation high throughput DNA sequencing to decode the transcriptomes and genomes of CAM species as a basis for discovering all of the genes required for efficient
CAM. We focus on genes involved in the optimisation of the biochemical steps of CAM over the light dark cycle, particularly on components of the circadian clock signalling pathway linking CAM biochemistry to the central molecular circadian clock. This talk will present an overview of our recent progress in understanding the functional genomics of CAM in our chosen biofuels feedstock crop, Agave sisalana .
cis-Golgi localization mechanism of type 2 prolyl 4hydroxylases; targeting domain and possible partner
Matsuoka, K 1,2 , Moriguchi, R 1 , Ohsawa, Y 2 , Suyama, A 1
1 Laboratory of Plant Nutrition, Faculty of Agriculture,
Kyushu University, Fukuoka, Japan;
Science Center, Yokohama, Japan
2 RIKEN Plant
Peptidyl prolyl 4-oxigenase, which is also called prolyl
4-hydroxylase (P4H), catalyzes the formation of hydroxyproline residues in proteins. Two classes of
P4Hs, type1 P4H that contain N-terminal signal anchor and type 2 P4H that contain both N-terminal signal peptide and C-terminal toxin homology domain (tox1 domain) has been found in plants. We cloned two full-

length cDNAs for type 2 P4H (NtP4H2.1, NtP4H2.2) from tobacco and analyzed the encoded proteins. Cell fractionation study using NtP4H2.2-specific antibody and the GFP fusion study indicated that NtP4H2.2 is localized predominantly in the cis-Golgi. Biochemical analysis revealed that NtP4H2.2 is a peripheral membrane protein tightly associated to the luminal side of microsoms. Expression of GFP-tox1 domain into the luminal part of the secretory pathway directed the GFP to cis-Golgi in both tobacco BY-2 cells and Arabidopsis leaf epidermal cells. Membrane fractionation analysis and brefeldin A treatment of tobacco cells expressing
GFP-tox1 also demonstrated that tox1 domain directs
GFP to the cis-Golgi. In addition, membrane association of GFP was enhanced by the fusion with tox1 domain.
Tox1 domains from both Arabidopsis and rice also directed GFP to the cis-Golgi in tobacco BY-2 cells.
These results indicate that tox1 domain is a novel cis-
Golgi targeting domain and its function is conserved in higher plants. In a separate experiment we searched interacting partner of NtP4H2.2 to address the localization mechanisms of this protein. Cross-linking experiment allowed us to identify a couple of possible partners. One of them corresponds to a tobacco ortholog of proteins that are involved in GPI-anchor remodeling.
This protein is of particular interest as many of the precursors to arabinogalactan protein contain GPIanchor. The tobacco cDNA for this protein was cloned and the characterization of this protein is under way. We observed that expression of this protein in yeast mutant complemented the phenotype and HA-tagged this protein in tobacco BY-2 cells was predominantly localized to the cis-Golgi. These observations raise a possibility that proline hydroxylation and subsequent glycosylation are somehow coupled with the remodeling of GPI-anchor during the biosynthesis of arabinogalactan proteins. This work was supported by a grant from Ministry of
Economy, Trade and Industry, Japan. We thank Drs.
Yoshifumi Jigami and Takehiko Yoko-o in National
Institute of Advanced Industrial Sciences and
Technology for yeast strains and expression vectors.
Identification and characterization of fucosyltransferases and galactosyltransferases involved in the biosynthesis of arabinogalactanproteins
Showalter, A 1
Kieliszewski, M
, Wu, Y 1
2 , Faik, A 1
, Williams, M 1 , Liang, Y 1 ,
1 Ohio University, Dept of Environmental & Plant
Biology, Molecular and Cellular Biology Program,
Athens, USA; 2 Ohio University, Dept of Chemistry and
Biochemistry, Molecular and Cellular Biology Program,
Athens, USA
Arabinogalactan-proteins (AGPs) are highly glycosylated hydroxyproline-rich glycoproteins (HRGPs) located at the cell surface and function in various aspects of plant growth and development. Arabinogalactan (AG) polysaccharides are attached to hydroxyproline (Hyp) residues in the protein backbone via a Hyp-Gal linkage and contain arabinose, galactose (Gal), rhamnose, glucuronic acid, galacturonic acid, and occasionally fucose (Fuc). Each protein backbone is decorated with numerous AG polysaccharides which are likely added in the endoplasmic reticulum and Golgi by specific glycosyltransferases. This presentation reports on two non-redundant Arabidopsis fucosyltransferases, FUT4 and FUT6, which add Fuc to AG polysaccharides and on the isolation and characterization of AGP-specific galactosyltransferase (GalT) activities. Multiple lines of evidence indicate that the AtFUT4 and AtFUT6 genes, members of the glycosyltransferase 37 family, encode alpha (1,2)fucosyltransferases (FUTs) for AGPs. First, overexpression of the two genes in tobacco BY2 cells, known to contain nonfucosylated AGPs, resulted in a staining of transgenic cells with eel lectin, which specifically binds to terminal alpha-linked fucose.
Second, monosaccharide analysis by high pH anion exchange chromatography and mass spectrometry indicated the presence of Fuc in AGPs from transgenic cell lines but not in AGPs from wild type cells. Third, detergent extracts from microsomal membranes prepared from transgenic lines were able to fucosylate purified
AGPs from BY2 wild type cells. Susceptibility of
[14C]fucosylated AGPs to alpha (1,2)fucosidase, and not to alpha (1,3/4)fucosidase, indicated that an alpha (1,2) linkage is formed. Furthermore, dearabinosylated AGPs were not substrate acceptors for these enzymes, indicating that arabinosyl residues represent fucosylation sites on these molecules. Testing of several polysaccharides, oligosaccharides, and glycoproteins as potential substrate acceptors in the fucosyl transfer reactions indicated that the two enzymes are specific for
AGPs but are not functionally redundant because they differentially fucosylate certain AGPs. AtFUT4 and
AtFUT6 are the first enzymes to be characterized for
AGP glycosylation. In addition, AGP GalT activities in tobacco and Arabidopsis microsomal membranes were studied with an in vitro GalT reaction system, which utilized acceptor substrates composed of [AO] repetitive units, specifically, a chemically synthesized [AO]7 acceptor and a transgenically produced and deglycosylated d[AO]51 acceptor. Incorporation of
[14C]Gal from UDP-[14C]Gal into the [AO]7 and d[AO]51 acceptors was observed following HPLC fractionation of the reaction products. Hyp-[14C]Gal monosaccharide and Hyp-[14C]Gal disaccharide were identified in the base hydrolysates of the GalT reaction products, indicating the presence of two distinct GalT activities for the addition of the first and second Gal residues to the [AO] peptide. Examination of the
Arabidopsis Hyp:GalT activity using various acceptor substrates, including two extensin sequences containing
SO4 modules and a [AP]7 peptide, indicated this activity was specific for peptidyl Hyp in AGP sequences. Mass spectrometry analysis demonstrated that only one Gal was added per peptide molecule to the C-terminal or pentultimate Hyp residue of the [AO]7 peptide. In addition, [AO]7:GalT and d[AO]51:GalT activities were localized to the endomembrane system of Arabidopsis suspension cultured cells following sucrose density gradient centrifugation. This in vitro GalT assay provides a useful tool for the identification and verification of
AGP-specific GalT proteins/genes.
Enzymes of CAZy GT-family-31: towards unravelling the function(s) of the candidate glycosyltransferase family members in
Arabidopsis thaliana
Egelund, J 1 , Ellis, M 2 , Doblin, M 2 , Fangel, JU 1 , Schultz,
C 3 , Willats, WGT 1 , Bacic, A 2
115

1 University of Copenhagen, Faculty of Life Sciences,
Section for Plant Glycobiology, Denmark; 2 Plant Cell
Biology Research Centre, School of Botany, The
3
University of Melbourne, Australia;
School of Agriculture, Food and Wine, The University of
Adelaide, Glen Osmond, Australia
Arabinogalactan-proteins (AGPs) are highly glycosylated macromolecules usually comprising 90-98% carbohydrate and 2-10% protein. They belong to a large family of plant cell wall (CW) glycoproteins, the hydroxyproline-rich glycoproteins (HRGPs), that are ubiquitous throughout the plant kingdom. AGP backbones are typically rich in hydroxyproline (Hyp), which are usually substituted by type II ArabinoGalactan
(AG) chains. AGP protein backbones are extensively modified in the endoplasmic reticulum/Golgi apparatus prior to secretion to the cell surface. This includes Oglycosylation by GTs (i.e. the addition of type II AG chains), and the addition of a glycosylphosphatidylinositol (GPI) anchor, that enables the attachment of AGPs to the plasma membrane. In addition to the important roles in plant growth and development, AGPs have numerous applications as functional food ingredients and are increasingly recognized as having health benefits. However, the biosynthetic machinery involved in the synthesis of the glycan component of AGPs is poorly understood. We will present an overview of the Arabidopsis thaliana members of CAZy GT-family-31 using our recent research as a working paradigm (Egelund et al., 2011;
Ellis et al., 2010; Qu et al., 2008). In addition, we will reveal the latest biochemical analysis of some of these putative galactosyltransferase (GalT) candidate genes responsible for synthesizing the beta(1,3)-Gal linkages of the type II AG chains.– a step in revealing the catalytic specificity of the Arabidopsis thaliana members of
CAZy GT-family-31.
Mutants with altered O-glycosylation levels of the cell wall protein extensin reveal a role in lateral root development
Gille, S 1 , Roycewicz, P 2 , Malamy, J 2 , Pauley, M 1
1 University of California, Berkeley, Plant and Microbial
Biology Dept, Berkeley, CA, USA; 2 The University of
Chicago, Molecular Genetics and Cell Biology, Chicago,
IL, USA
Plant cells are encased in a cell wall that plays multiple roles in the life cycle of a plant including the maintenance of structural integrity and a conduit for environment to cell signals. Cell walls consist of various polymer networks mainly polysaccharides (cellulose, various hemicelluloses, and pectins) and the polyphenol lignin, but also a glycoprotein network. One major component of this proteinaceous network is extensin, a protein containing a large abundance of hydroxyprolineresidues, which are O-glycosylated with arabinosyl- and to a lesser extend galcatosyl-residues. Proper glycosylation of extensin has been shown to be important for its conformation and polymerization capability, but the physiological role of extensin glycosylation remained elusive. In a forward genetic screen we have identified an
Arabidopsis mutant with a lack of a transcript encoding
XEG113, a CAZy family 77 glycosyltransferase protein.
116
Analysis of the cell wall of the mutant revealed a severe reduction in extensin glycosylation suggesting that
XEG113 represents an extensin:arabinosyltransferase. As a result of extensin underglycosylation mutant plants have a larger appearance and elongated hypocotyls. In addition, under certain conditions lateral roots appeare in the mutants, while wildtype plants lack those. We hypothesize that underglycosylation of extensins lead to a disruption of the wall glycoprotein network, which in turn facilitates the emergence of the lateral roots through the root cortex tissue. Moreover, we identified a knockout mutant of a close homolog, XEG113-like.
Characterization of this mutant and the double mutant
XEG113/ XEG113-like suggests that XEG113-like also represents an arabinosyltransferase and acts additive to
XEG113. The physiological role of XEG113-like is currently investigated and will be discussed including its synergies with XEG113.
Identification of a novel glycosyltranseferase gene
1 family in plants
Shimma, Y-I 1
T 1
, Saito, F 1 , Suyama, A 2
, Matsuoka, K 2 , Jigami, Y 1
, Oka, T 3 , Yoko-o,
Research Center for Medical Glycoscience, National
Institute of Advanced Industrial Science and Technology
2
(AIST), AIST Tsukuba Central 6, Tsukuba, Japan;
Laboratory of Plant Nutrition, Faculty of Agriculture,
3 Kyushu University, Fukuoka, Japan; Dept of Applied
Microbial Technology, Sojo University, Kumamoto,
Japan
Plant glycoproteins such as extensin and arabinogalactan proteins are modified with plant specific O-linked galactosylation on serine and hydroxyproline (Hyp) residues. An assay system for peptidyl serine Ogalactosyltransferase (SGT) activity was established, using UDP-galactose as a donor, and chemically synthesized peptides of extensin-like amino acid sequences as an acceptor. The neighboring Hyp residues adjacent to serine in the acceptor peptide were required for SGT activity. SGT protein was purified from cell extracts of C. reinhardtii , and subjected to Mascot analysis of proteins to determine the amino acid sequences by mass spectrometry. The analyzed peptide sequences were completely matched with an ORF in the
C. reinhardtii genome database. A corresponding DNA fragment was cloned from C. reinhardtii cDNA library.
The SGT activity was confirmed through the cell surface expression system in yeast Saccharomyces cerevisiae, therefore, designated as CrSGT1. Highly homologous
ORFs were found in various plant genomes, such as
Arabidopsis thaliana , Nicotiana tabacum, Oryza sativa , etc. The AtSGT1 and NtSGT1 also showed SGT activity when expressed in yeasts. The SGT proteins contained a conserved DXD motif and showed a type I membrane topology, but no homology with known glycosyltransferases, indicating that SGT1 is a novel glycosyltransferase gene family, existing only in plant kingdom.

Complexity and coordination of root growth under water deficit: recent advances from transcriptomic and cell wall proteomic analyses
Sharp, RE
Bunzel, M 2
1 , Voothuluru, P
Byro-Jilek, M 2
1 , Yamaguchi, M 1 , Zhu, J 1
, Garnett, JS
, Simmonds, J 4
1 , Cho, I-J 3
, Wu, Y 5
, Oliver, MJ 3
,
,
1 University of Missouri, Columbia, USA;
Minnesota, St Paul, USA;
4
3
2 University of
USDA-ARS, Columbia,
Missouri, USA;
Ottawa, Canada;
Agriculture and Agri-Food Canada,
5 South Dakota State University,
Brookings, USA
Previous work on maize primary root adaptation to water deficit showed that the response of cell elongation varies at different positions within the growth zone. In the apical 3 mm (region 1), elongation is maintained at wellwatered rates even under severe stress (water potential of
-1.6 MPa), whereas in the 3–7 mm region (region 2), which exhibits maximum elongation in well-watered roots, elongation is progressively inhibited in stressed roots. In association with these responses, cell wall extensibility is enhanced in the apical region but decreased in the basal region. This presentation will focus on recent advances in understanding of these responses from transcriptomic and cell wall proteomic analyses. Cell wall proteomic analysis (Zhu et al. 2007) revealed region-specific changes in protein profiles between well-watered and water-stressed roots. In particular, several proteins related to reactive oxygen species (ROS) generation increased in abundance particularly in region 1, prominent among them being putative oxalate oxidases (OxO), which produce hydrogen peroxide. To investigate the role of
OxO/apoplastic ROS in root elongation, we have characterized transgenic maize lines constitutively expressing a wheat OxO gene. Results show that OxO activity is increased in a region-specific manner with different profiles in well-watered and water-stressed roots, and that these changes are associated with differential effects on growth and growth-related processes. Interestingly, the results indicate that cell production rather than cell expansion processes are altered in the OxO transgenic lines. Previous work showed that in both regions 1 and 2, the activity of wallloosening expansin proteins is enhanced in water-stressed compared with well-watered roots. In region 1, the wall susceptibility to expansins is also increased, whereas in region 2, expansin-induced wall extension is inhibited.
Ferulates are abundant in the cell walls of monocotyledonous plants and have a role in cross-linking wall polysaccharides and other components. Microarray analysis suggested that differential regulation of ferulate metabolism between the apical and basal regions of the root growth zone could contribute to the different changes in wall extensibility and expansin susceptibility under water stress. To test this hypothesis, the spatial distribution of wall-bound ferulates was determined by histochemical and HPLC analyses. The results show that ferulate content decreases and increases, respectively, in regions 1 and 2 of water-stressed compared with wellwatered roots. To evaluate whether the accumulation of ferulates in region 2 prohibits the ability of expansins to extend the cell walls, root segments were treated with feruloyl esterase (FE), which can release ferulates from a range of esterified substrates. An extensometer system was used to show that FE treatment completely restores expansin-induced wall extension in region 2 of waterstressed roots, while having a minimal effect in the same region of well-watered roots. These studies provide novel insights into the complexity and coordination of root growth regulation and adaptation under water deficit conditions. This research was supported in part by a grant to R.E.S. from Monsanto.
The Plant Ontology: an updated tool for all plant biologists
Walls, RL 1 , Cooper, L
Mungall, C
Jaiswal, P 2
4 , Justin, P 2
2 , Elser, J
, Smith, B 5
2 , Gandolfo, MA 3
, Stevenson, DW 1
1 New York Botanical Garden, USA;
University, USA; 3
2 Oregon State
Cornell University, USA; 4 Lawrence
Berkeley National Lab, USA; 5 University at Buffalo, USA
The Plant Ontology (PO: http://plantontology.org) is a
,
, structured vocabulary and database resource for plant researchers that links plant anatomy and development to the rapidly expanding field of genomics in the domain of all green plants. The PO contains interrelated concepts or terms, organized in a hierarchical tree, that describe plant anatomy and morphology, as well as plant growth and development stages. Like all ontologies, the PO facilitates data sharing and automated analyses through the consistent use of vocabulary with explicit, logical definitions and defined relationships among terms.
Originally designed to span the monocot-dicot divide in flowering plants (based on the model angiosperms Zea mays , Oryza sativa , and Arabidopsis thaliana ), the PO has been redesigned to encompass all angiosperm species and to include terms and annotations from non-flowering plants such as gymnosperms, pteridophytes, and bryophytes. This is the first ontology designed to encompass anatomical structures as well as growth and developmental stages across such a broad taxonomic range. Over the past 18 months, upper level parent terms have been added, renamed, or revised to ensure applicability to the entire plant kingdom, allowing comparative studies across distantly related taxa. At the same time, many new clade-specific terms have been introduced at lower levels, allowing precise annotation of genomic and genetic datasets for individual species.
Throughout the revision process, curators worked to incorporate advances in ontology design and construction. These included the use of logical text and relational definitions and references to external ontologies where appropriate. An essential, powerful feature of the PO is the links through terms to associated structure or growth stage specific genes, proteins, and phenotypes from numerous genomics datasets. One of the main goals of the PO is to facilitate cross-database querying and to foster consistent use of vocabularies in annotation. To this end, the PO is developing annotation tools for the plant science community. Outreach activities include annotation workshops and outreach booths and symposia at international conferences. Currently, the PO includes over 500,000 annotations associated with over
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1,100 terms. In this capacity, it is widely applicable to plant genomic and evolutionary developmental studies.
In addition, image libraries are being created through
PlantSystematics.org and linked to terms to provide reference images for plant structures, along with the definitions. This makes the PO a valuable resource for research and teaching that can be used as a guide to plant structures and the common growth and developmental landmarks in lifecycles of plants across taxa. This presentation will provide an overview of the Plant
Ontology and the annotation database, as well as potential applications of the PO. The PO is publicly available and collaboration with diverse user groups is encouraged. Information will be available for research groups that wish to provide annotations or use the PO in novel applications.
Unraveling Darwin’s ‘abominable mystery’: using phylogenomics to identify proteins of importance in plant evolution
Cibrian, A 1
G 4
, Martienssen, R 2
, Desalle, R 5
, Stevenson, D 3 , Coruzzi,
1 Center for Genomics and Systems Biology, New York
University, USA; 2 Cold Spring Harbor Laboratory, USA;
3 The New York Botanical Garden, USA; 4 Center for
Genomics and Systems Biology, New York University,
USA; 5 American Museum of Natural History, USA
Darwin was famously concerned that the sudden appearance and rapid diversification of flowering plants in the mid-Cretaceous could not have occurred by gradual change. Here I present a phylogeny among the major seed plant groups, i.e. cycads, ginkgo, conifers, gnetophytes, and flowering plants based on complete genomes and unigenes from 150 plant species. This is the largest plant phylogenomic matrix to date. I provide a bioinformatic pipeline in which these phylogenetic relationships can be used as a platform for identifying genes of functional importance in plant diversification.
Genes with positive partitioned Bremer support at major nodes were used to identify overrepresented gene ontology (GO) terms. This is a novel method in which phylogenomic tools are used to postulate hypotheses of gene function in the evolution of major taxa. Although I focus on plants, it can be applied to any group of species with ESTs or genomes available. We provide support for one of the long-standing, most controversial hypothesis of gymnosperm evolution. We also found that genes involved in PTGS (post-transcriptional gene silencing) provide support for the divergence of the monocots in early angiosperm evolution. One of these genes, RNAdependent RNA polymerase 6, is required for the biogenesis of trans-acting small interfering RNA
(tasiRNA), confers heteroblasty and organ polarity, and restricts maternal specification of the germline.
Functional hypotheses such as the role of these genes for plant diversification can be further coupled with expression and genetic data but their phylogenetic importance is identified with our approach first. This can lead to better gene searches, annotations, and functional analysis of genome level studies. This phylogenomic approach and those genes identified here guide plant ecological genomics studies and help understand the precise evolutionary mechanisms driving the diversification of plant species, gradually unraveling
Darwin’s abiding and perplexing mystery.
Investigating trait ontologies to facilitate integrating phenotype and genome sequence level information in
1 wheat
Appels, R 1
Centre for Comparative Genomics, Murdoch University,
Perth, WA, Australia
Commercially important crops such as wheat ( Triticum aestivum L.) have an extensive history of data collection, relating phenotype to genetic information. The catalogue of gene symbols for wheat
(http://www.shigen.nig.ac.jp/wheat/komugi/genes/symbo lClassList.jsp, edited by R.A. McIntosh), provides a well- (meaning manually) curated assessment of the genetic studies carried out to map traits of interest to genetic maps. These traits are assigned symbols, and an example of an entry is: Pre-harvest sprouting QTL:
Several QTL for falling number and alpha-amylase activity, two indicators for pre-harvest sprouting resistance, were identified in {0169}. The most significant were associated with Xglk699-2A and
Xsfr4(NBS)-2A, Xglk80-3A and Xpsr1054-3A,
Xpsr1194-5A and Xpsr918-5A, Xpsr644-5A and
Xpsr945-5A, Xpsr8(Cxp3)-6A and Xpsr563-6A, and
Xpsr350-7B and Xbzh232(Tha)-7B {0169}. Typically, a trait such as pre-harvest sprouting will have several synonyms, such as sprouting index (QSi.crc-5D), dormancy (Q.SD1), or pre-harvest sprouting (Qphs.ocs-
3A.2), which need to be captured for an effective analysis of published information. In order to crossreference genome sequence information to trait/phenotype studies, the available information in wheat is now cross-referenced to detailed consensus molecular genetic
(http://ccg.murdoch.edu.au/cmap/ccg-live/cgimaps bin/cmap/viewer) in the CMap software utilized by
GRAMENE (http://www.gramene.org/). In this presentation, the experience of developing trait ontologies for wheat, using the information established for rice, will be discussed in the context of relating the
CMap-based information that locates published QTL for traits to the genome sequence of wheat. The sequence of a complex genome such as wheat has been challenging to develop, but several new technologies are now converging to establish draft sequence-level definitions of the gene-space.
The past, present and future: ontologies for plant
1 biology
Jaiswal, P 1 , Cooper, L 1 , Elser, J
Oregon State University, USA
1
As a result of current developments in sequencing technology, the number of sequenced plant genomes is rapidly increasing. Along with the deluge of information on nucleotide and amino acid sequences from a genetically diverse set of species, large quantities of data are also being generated on genes, alleles, their polymorphism, protein functions, transcriptome, proteome and metabolomics experiments, mutants and
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their phenotypes, metabolic and regulatory interaction networks, and literature references. In the midst of this influx of data, we also realize that few species are being studied in sufficient detail to understand how plants develop, adapt, and diversify. Therefore, experimental annotations generated for a set of well-studied species are being increasingly used to project annotations for lesserstudied species. This demands that the annotations from the well-studied species be generated meeting current genomics/genetics standards and remain up to date all the times. It is, however, time-consuming and laborious for individual researchers to collect information from multiple original data sets and to rearrange it for their own purposes. The concept of ontology has been introduced to biology to support and encourage researchers to share and reuse information among biological databases to foster consistency and use of common vocabulary. Ontology is a kind of glossary that has a built-in hierarchical structure, logical definitions, and relationships among concepts, and is used to define biological processes, functions and environments from the cellular to the organismal level. In the presentation, I will introduce ontology-based annotation strategies being adopted by the new generation of plant genomics and phenomics projects and their role in the discovery environment.
PODD: an ontology driven architecture for extensible phenomics data management
Kennedy, G
B 1
1,2 , Li, Y-F 2 , Davies, F 2 , Hunter, J 1 , Furbank,
1 The Australian Plant Phenomics Facility, High
Resolution Plant Phenomics Centre, CSIRO, Canberra,
2 Australia; School of ITEE, The University of
Queensland, Brisbane, Australia
Ontologies have found increasing favour in the plant sciences because they can deliver a set of terminologies and understandings about biological concepts that are agreed between researchers. Typically ontologies are used to annotate data on the web, but the notion of a common vocabulary with formally defined semantics makes ontologies the vehicles for representing data and knowledge in the Semantic Web. Ontologies provide unambiguous classifiers and descriptors that are made available in a format other computers can autonomously discover and interrogate, and thus may be linked across disparate databases and repositories. In the Phenomics
Ontology Driven Data repository (PODD) we have taken the notion of classification of experimental concepts using ontologies one step further by using an ontology, the PODD ontology, as the schema of our data management system. We utilise the Semantic Web ontology languages OWL and RDFS to do this because they provide the extensibility and the semantic rigour required. In this ontology-driven architecture the behaviours of domain concepts and objects are captured entirely by ontological entities, around which all data management tasks are carried out. An ideal domain for applying these principles is plant phenomics, the systematic study of the phenotypes of model and crop plants that are a consequence of the individual plant's genome and environment. Phenomics research generates high volumes of heterogeneous data through the use of emerging imaging and measurement technologies and processes. This data is combined with metadata to form complex digital objects and then further associated with provenance metadata on the experimental process. In this context, we describe the development of a phenomics experimental process ontology, and how we have applied the principles of ontology-driven architecture in the development of PODD, a data management system for phenomics based research. Keywords: Phenomics,
Semantic Web, Ontologies, OWL, data management,
PODD
Predicted protein–protein interactions in the moss
Physcomitrella patens
: a new bioinformatic resource
Schuette, S 1 , Corley, A 1 , Lang, D 2 , Geisler, M 1
1 Dept of Plant Biology, Southern Illinois University,
Carbondale IL, USA; 2 Plant Biotechnology, University of
Freiburg, Freiburg, Germany
Physcomitrella patens , the model moss for plant biology, has a protein-coding genome similar in size to
Arabidopsis , but is similar to yeast in efficiency of gene targeting experiments and has a haploid dominant form making an interesting and useful molecular genetic tool for plants. The availability of the moss genome has made possible the exploration of plant diversity at the molecular level. The model moss is fast becoming a tool for bioinformatic and molecular work due to its key phylogenetic position as sister to land plant lineages. We present here the first predicted protein-protein PPI for a bryophyte based on the interolog method. Whole genome sequences from reference species including yeast, nematode worm, fruitfly, mouse, rat, human, bacteria and
Arabidopsis were compared to the genome of
Physcomitrella patens in a pairwise fashion using reciprocal blasts to separate inparalogs from orthologs and outparalogs with INPARANOID software package.
A reference interaction database was assembled using
MySQL by compiling BioGrid, BIND, DIP, and Intact databases. The reference database was queried for which moss orthologs existed for both interacting partners. We predicted more than 60 000 total interactions from different predicting references including 41,936 unique interactions from 4062 different P. patens proteins that were visualized in Cytoscape, a Java-based software package. The twenty most interactive proteins represent strongly conserved pathways that have not altered significantly during eukaryotic evolution. Analysis of gene ontology revealed the most significant categories represented include metabolic processes, intracellular and cytoplasmic likely due to their conserved nature, and protein binding due to physical interaction requirement for inclusion, and catalytic activities. The utility of predicted interactomes lies in the 'guilt-by-association' model of predicting proteins in a pathway under the assumption that orthologous proteins have similar functions. For example, we constructed a Calvin Cycle network to determine the number of proteins associated with this all-important process and discovered an uncharacterized protein with phosphoglycerate kinase activity that interacts directly with NADP-ME, an enzyme involved in C photosynthesis. The addition of
4 moss, a plant representative 200 million years diverged from Arabidopsis , to interactomic research greatly expands the possibility of conducting comparative analyses thus giving tremendous insight into network
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evolution of land plants.
Exploring natural epigenetic variation and its implications for plant evolution
Paun, O 1 , Stuessy, TF 1 , Chase, MW 2
1 Dept of Systematics and Evolutionary Botany,
University of Vienna, Austria; 2 Jodrell Laboratory,
Royal Botanic Gardens Kew, UK
Without altering the underlying DNA sequence, epigenetic information influences the identity of cells and their response to the external and internal environment by modulating gene expression and regulating repetitive elements. Epigenetic changes can be transient, being reset between generations. However, several spontaneous or induced epigenetic variants were shown to be stably inherited across multiple generations. Our laboratorybased mechanistic understanding of epigenetics in model organisms is expanding rapidly. These exciting new findings indicate that divergent epialleles influence phenotypic features and have the potential to affect adaptation and divergence, even in the absence of genetic variability. We have now the first evidence that natural selection acts on epigenetic variation in a similar way as on genetic components and results in novel adaptations.
Epialleles, like random genetic mutations, can alter development and metabolism and therefore have evolutionary implications. However, epigenetic variation is often under direct environmental influence and may be generated at a much higher rate than genetic diversity, especially in rapidly changing environmental conditions.
Different degrees of epigenetic silencing or activation can further produce continuous variation in phenotypes rather than discrete phenotypic classes. Environmentally stimulated disruption of epigenetic control may also simultaneously induce several new epimutations in the same individual, in marked contrast with random spontaneous genetic changes. The tools necessary for epigenetic investigations are already available even for non-model organisms, but so far they have been insufficiently explored to study this level of natural variation. In order to improve the general understanding of natural phenotypic variation and organismic adaptive strategies, we need to complement the gene- and genome-centered view, currently dominating our evolutionary perspective about variability, heritability and adaptation, with a substantial consideration of epigenetic factors.
Trans-generational inheritance in flax – association with genomic re-arrangements and small RNAs
Cullis, C 1 , Johnson, C 1 , Moss, T 1
1 Case Western Reserve University,USA
Flax ( Linum usitatissimum ) can undergo heritable changes in phenotype and genotype in response to the growth environment. These changes can be stable over many generations or be reversible following alterations
120 in the growth environment in subsequent generations.
The responses have the characteristics of epigenetic modifications. They are also associated with reversible genomic reorganization events including the novel insertion event, named LIS-1, which can arise in flax in response to particular growth conditions. LIS-1 was observed to always become both homozygous and inherited in subsequent generations under a particular growth environment, sporadically appeared but was never transmitted to the progeny under another growth environment, while has not been observed either during growth or in the next generation under a third growth environment. In some flax varieties that did not produce stable genotrophs, LIS-1 was subsequently lost unless the appropriate growth environment was maintained. Thus appearance and subsequent inheritance of LIS-1 is specific to particular growth environments and is not part of the normal developmental program of flax. Stable lines (genotrophs) resulting from the initial growth under each of these conditions appear to grow better when grown under the same conditions in subsequent generations suggesting that adaptive changes have taken place. The ability of flax to respond to the environmental conditions (as assayed by the appearance of LIS-1) is genetically controlled. The loci controlling the ability to respond to the growth environment have been mapped in
F2 populations of crosses between responsive and stable lines. and candidate genes being identified.
Responsiveness is dominant over stability. LIS-1 is the result of a precise programmed set of events in response to the growth environment although its origins remain unclear. The element is not present in an intact form in responsive lines prior to its appearance. However, primers spanning putative small RNA encoding segments within LIS-1 can be used to amplify, from DNA isolated from varieties lacking LIS-1, stretches of sequences similar to, and sometimes identical to, the final LIS-1 element. One possible analogy to this novel molecular restructuring process in flax is the series of molecular events that accompany the ciliate macronuclear development which are facilitated by guiding by micro
RNAs. Although it is accepted that small RNAs are capable of causing epigenetic regulation of genes through modulation of chromatin, this system in flax offers the opportunity to determine the involvement of small RNAs in reversible, physical genomic rearrangements induced by environmental stress that may also affect gene expression and possible adaptation. The specificity of the appearance and subsequent inheritance of LIS-1 and associated phenotypes to particular growth environments demonstrates that the environment can act as both the inducer of targeted genetic variation and as the selective agent for advantageous mutations. The evolutionary implications of this mechanism for any organism in which the germline is not set aside very early in development is that a selectable, coordinated set of variations can arise under particular environmental stresses resulting rapid adaptive evolutionary responses.
Epigenetic and genetic variation in imprinted genes between
Arabidopsis thaliana
and
Arabidopsis lyrata
Fujimoto, R 1 , Sasaki, T 2 , Dennis, E 1
1 CSIRO Plant Industry, Canberra, Australia; 2 Gregor
Mendel Institute of Molecular Plant Biology, Vienna,
Austria

DNA methylation is one of the epigenetic marks associated with transposable element silencing and parent of origin-specific gene expression (imprinting) in plants and mammals. In plants, imprinted gene expression occurs in the endosperm by cytosine demethylation of the maternal genome mediated by the DNA demethylase
DEMETER. In Arabidopsis thaliana , imprinted genes often contain invading sequences such as transposons in their flanking regions, and it has been suggested that imprinting evolved from targeted methylation of transposable element. FWA gene is an imprinted genes and silencing of FWA in vegetative tissues is dependent on cytosine methylation in the region of a SINE retroelement. We examined the critical region for DNA methylation in both A. thaliana and A. lyrata using double stranded RNA to direct DNA methylation to target regions. DNA methylation in the region upstream from the transcription start site plays a role in FWA silencing in both species, while DNA methylation in the region downstream from the transcription start site is important for the FWA silencing only in A. thaliana .
Large tandem repeats are present in this region including the transcription start site only in A. thaliana . These results indicate that the acquisition of the large tandem repeats enlarged the critical methylated region in A. thaliana . We also examined DNA methylation of the promoter region of other genes in A. lyrata which are imprinted in A. thaliana . We found both conservation and variation of the methylated regions of these genes between A. lyrata and A. thaliana .
Chromatin in signaling of plant development and response to environmental stress
Dong, A 1
W-H 1
, Zhu, Y 1 , Yu, Y 1 , Berr, A 1 , Ménard, R 1 , Shen,
1 State Key Laboratory of Genetic Engineering, Dept of
Biochemistry, Institute of Plant Biology, School of Life
Sciences, Fudan University, Shanghai, China; Institut de
Biologie Moléculaire des Plantes du CNRS, Université de Strasbourg, France
Chromatin represents the physiological template of genetic information in all eukaryotes. The basic unit of chromatin is the nucleosome, which is composed of 146 base pairs of DNA wrapped around an octamer of two molecules of each of the histones H2A, H2B, H3, and
H4. Nucleosome assembly/disassembly is mediated by histone chaperones and occurs during DNA replication as well as during gene transcription, DNA repair, and recombination. Our work on Arabidopsis histone chaperones has demonstrated that proper nucleosome assembly is crucial for maintenance of genome integrity, cell proliferation and plant response to genotoxic stresses
(1–3). Histone methylation is one type of the epigenetic marks that play essential regulatory functions in chromatin structure organization and genome function.
Our work on Arabidopsis SET-domain histonemethyltransferases has shown that histone H3 lysine 4
(H3K4) and H3K36 methylations are critical for transcriptional activation of several key regulatory genes involved in flowering time control or plant reproduction
(4–7). Remarkably, H3K36 methylation is also dynamically involved in plant defense against fungal pathogens (8). We found that the Arabidopsis PRC1-like ring-finger proteins, which likely read the H3K27 trimethylation, are necessary for repression of embryonic traits and for maintenance of proper stem cell activity during plant vegetative growth (9, 10). We will present and discuss our most recent progress in understanding of nucleosome assembly and histone code as epigenetic mechanisms in signaling of plant development and in plant response to environmental cues. (1) Zhu et al. 2006.
(2) Liu et al. 2009. (3) Liu et al. 2009. (4) Zhao et al.
2005. (5) Xu et al. 2008. (6) Berr et al. 2009. (7) Berr et al. 2010. (8) Berr et al. 2010. (9) Xu and Shen 2008. (10)
Chen et al. 2010.
Epigenetic regulation of pollinator syndromes
Baumberger, R 1,2 , Grossniklaus, U 1
2
1 Institute of Plant Sciences & Zürich-Basel Plant Science
Center, University of Zürich, Zürich, Switzerland;
Pädagogische Hochschule Zürich, Zürich, Switzerland
Until recently, epigenetic variation was not thought to play any significant role in evolution and most text books on evolutionary biology lack any reference to epigenetic processes. Indeed, many biologists have difficulties to see how epigenetic variation could contribute to evolutionary change, as exemplified by the statement:
‘… it is hard to see its [epigenetics] possible evolutionary significance, …’ (1). Nevertheless, over the last few years epigenetic research has taken centre stage and several scientists have argued for a role of epigenetic variation in evolution (2–4). A few years ago, we proposed that apomixis, the asexual reproduction through seeds, may have evolved through epigenetic changes caused by inter-specific hybridization and polyploidization (5, 6), but apomictic systems proved difficult to study in this respect. Here, we present a new model system that may allow us to analyze epigenetic changes and their possible role in evolution at the genetic and molecular level. We will provide evidence that different taxa of Mimulus spp. with distinct pollinator syndromes are in fact epigenetic variants. An insectpollinated plant with yellow flowers can change, over the course of several years, into a plant with bird-pollinated, red flowers. Moreover, this epigenetic state is heritable and behaves like an epimutation. Since the acquired epigenetic state affects the morphology, colour, and scent of the flower and leads to reproductive isolation, it is expected to have a strong effect on population structure and eventually the evolutionary trajectory of this taxon.
(1) Wolpert L 1998. (2) Jablonka E, Lamb MJ 1995. (3)
Rapp RA, Wendel JF 2005. (4) Richards EJ 2006. (5)
Grossniklaus U et al. 1998. (6) Spillane C et al. U 2001.
Genomic consequences of hybridisation and polyploidy in
Senecio
Hegarty, M
Abbott, R 3
1 , Barker, G
, Edwards, K 2
2 , Brennan, A
, Hiscock, S 2
3 , Batstone, T 2 ,
2
1 Aberystwyth University, IBERS, Aberystwyth, UK;
University of Bristol, School of Biological Sciences,
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Bristol, UK; 3 University of St Andrews, School of
Biology, St Andrews, UK
Interspecific hybridisation (often involving a change in ploidy) is an important mechanism by which speciation can occur in plants. To better understand the impact of merging two divergent genomes within a single hybrid nucleus, we conducted a comparative transcriptomics assay in the genus Senecio , which provides examples of both diploid and allopolyploid hybrids formed within the last 300 years. These hybrids are recognised as examples of hybrid speciation, forming either in allopatry or sympatry. Using a custom microarray platform, we identified widespread non-additive changes to gene expression in both types of hybrid, and demonstrated that hybridisation and genome duplication have separate, distinct effects on gene expression. Subsequent research has focussed on investigating the epigenetic factors underpinning these expression changes and has shown that hybrids can display global modifications to DNA methylation in a nonadditive manner similar to that observed for gene expression. Allopolyploid Senecio hybrids have also been demonstrated to display biased expression of parental homoeologues. Current efforts are focused on the development of a draft reference genome for the diploid hybrid Senecio squalidus and the application of comparative genomics techniques to address outstanding questions about the consequences of genomic mergers.
Genetic, karyotypic and genomic evolution in
Tragopogon
allotetraploids
Soltis, DE
Chester, M 1
Muniz, A
Schnable, PS 5
WB 1
1
1 , Soltis, PS 1
, J. A. Tate 4
, Buggs, RJA 2
, Jordon-Thaden, IE 1
, Viccini, LF 3 ,
, Veruska Cruz da Silva
, Symonds, VV
, Chamala, S 1 , Davenport, R 1
4 , Wu W 5 ,
, Barbazuk,
1 University of Florida, USA;
London, UK;
Brazil; 4
3
2 Queen Mary University of
Universidade Federal de Juiz de Fora,
Massey University, New Zealand;
University, USA
5 Iowa State
Understanding polyploid evolution is central to understanding the origin and diversification of most lineages of life, particularly flowering plants. However, despite the prevalence and importance of polyploidy in plants, most of what we know about the genetic and genomic consequences of genome doubling is derived from synthetic polyploids, crops, and model organisms that are ancient polyploids (e.g., Arabidopsis ). To understand better how polyploidy shapes genome evolution and gene function, we have been investigating naturally occurring polyploids in the genus Tragopogon
(Asteraceae). Tragopogon provides two well-known examples of recent allopolyploid speciation and a unique opportunity to investigate the genetic and genomic changes that occur across a continuum from F1 hybrids, synthetic polyploids, and recently and recurrently formed natural populations of allopolyploids ( T. mirus and T. miscellus , each of which formed fewer than 80 years ago; i.e., 40 generations in these biennial plants). Also, this system provides the unusual opportunity to determine whether genome evolution follows a program and if underlying 'rules' govern genomic interactions in polyploids. We report patterns of extensive homeolog
122 loss, karyotypic variation, and shifts in gene expression across populations of both allotetraploid species. Recent advances in high-throughput sequencing technology provide a rapid and cost-effective means of generating sequence data. Given that Tragopogon is an excellent natural model for polyploid evolution, and our ready access to state-of-the-art sequencing technologies, we have assembled a draft genome sequence of the diploid,
Tragopogon dubius , which will provide a foundation for further analysis of the genetic and genomic consequences of allopolyploidy in Tragopogon .
The impact of paleopolyploidy on network evolution in the Brassicales
Edger, P 1
M 2
, Bekaert, M 1
, Jiao, Y 3
, Conant, G 1
, dePamphilis, C
, Hudson, C
3 , Schranz, E 4
1 , Barker,
, Pires, JC 1
1 University of Missouri – Columbia, USA;
Arizona, USA; 3
2 University of
Penn State University, USA; 4 University of Amsterdam, The Netherlands
Ancient whole genome duplications (WGDs), inferred from analyzed genomes and transcriptomes, are prevalent and recurring throughout the evolutionary history of higher eukaryotic lineages. The impact of these events on gene networks (e.g. metabolic, regulatory, signaling, and protein-protein interaction) remains unclear. Despite its small size (157Mb), the Arabidopsis thaliana genome shares the remnants of at least three ancient WGDs. We have constructed a phylogenomic framework, which includes having localized the two most recent WGD events (At-
α
and At-
β
), to investigate patterns of duplicate gene retention (i.e. gene family size evolution), innovation of novel gene functions, and the origin of novel pathways. In addition, we analyzed patterns of surviving duplicates from both At-
α
and At-
β
across the entire Arabidopsis metabolic network. These results has provided insights into the selective constraints that retain specific functional genes in duplicate post-WGD over millions of years, which over longer time periods (once constraints are lifted) may sub- or neo-functionalize.
Allopolyploid genome divergence is associated with genome downsizing and elimination of paternally derived repetitive DNA sequences in tobacco
Leitch, AR
A 3
1 , Renny-Byfield, S 1
, Le Comber, SC 1 , Grandbastien, M-A 4
, Macas, J 5
, Chester, M
, Novak, P
0 , Kovarík,
, Deloger, M
5 , Chase, MW 6
4 ,
1
Nicholas, RA 1
Queen Mary University of London, School of Biological and Chemical Sciences, London, UK; 2 Laboratory of
Molecular Systematics and Evolutionary Genetics,
Florida Museum of Natural History, University of
Florida, Gainesville, FL, USA; 3 Institute of Biophysics,
Academy of Sciences of The Czech Republic, Brno, Czech
Republic; 4 Institute Jean-Pierre Bourgin, INRA-
5 Versailles, France; Biology Centre ASCR, Institute of
Plant Molecular Biology, Czech Republic; 6 Jodrell Lab,
Royal Botanic Gardens, Kew, Richmond, Surrey, UK
Angiosperm evolution is heavily impacted by polyploidy, which has occurred in the ancestry of all, or most, species. Interspecific hybridisation combined with genome multiplication (allopolyploidy) provides a natural experiment in genome perturbation. Genetic

changes, subsequent to polyploid formation, can be inferred by studying the descendants of the two progenitor species and the allopolyploid offspring. In
1984 McClintock first proposed that these changes might be distinctive: that allopolyploidy can induce 'genomic shock', which may produce rapid alteration at the DNA sequence level in addition to significant structural, transcriptional, epigenetic and karyotypic changes.
Moreover such dynamism has been observed to occur extraordinarily rapidly in some model species, occurring after only a few generations. This has led many to envisage a 'genome revolution' where perturbation of progenitor genomes is induced by their unification. To investigate this phenomenon we used next generation sequencing to characterise and compare the genomes of the recently derived allotetraploid, Nicotiana tabacum
(tobacco, < 200,000 years old), with its diploid progenitors, N. sylvestris and N. tomentosiformis . We show that all major types of retroelements found in angiosperms are present. Sequence similarity comparisons and estimates of repetitive DNA abundance all suggest that the three genomes have experienced different evolutionary histories. The diploid N. sylvestris genome exhibits evidence of recent bursts of sequence amplification and/or homogenisation, whilst the genome of N. tomentosiformis reveals only hints of such a pattern. In contrast, the genome of tobacco shows evidence of sequence loss, particularly evident among the
Ty3-gypsy retroelements, but also involving tandem repeats and 35S ribosomal DNA. Moreover the paternally ( N. tomentosiformis ) derived T-genome of tobacco shows evidence of erosion, while the maternal
( N. sylvestris ) S-genome appears materially unchanged, indicating the possibility of preferential loss of paternally derived repetitive DNAs. These observations together are in-line with the genome downsizing and nuclear cytoplasmic interaction hypotheses. In a broad sense, we can consider the genomes of N. sylvestris, N. tomentosiformis and N. tabacum to be dynamic, stable and decaying, respectively.
Reconstruction of the chloroplast genome of
Spartina maritima
, and molecular dating of the divergence between the two genomes reunited in the recent allopolyploid
S. anglica
Bellott, S
Lima, O 1
1,2 , Ainouche, K
, Naquin, D
1 , Chelaifa, H
1 , Ainouche, M 1
1 , Coudouel, S 1 ,
1 Ecosystems, Biodiversity and Evolution, University of
Rennes, France; 2 Systematic Botany and Mycology,
University of Munich (LMU), Germany
The genus Spartina (Poaceae, Chloridoideae) is an important system for analyzing reticulate evolution and recurrent polyploid speciation. Eight of the 14 species are tetraploid (4x) and five are hexaploid (6x), among them some give rise to further allopolyploids (7x, 8x, 9x, 12x).
The birth of S. anglica c. 1890 by such hybridization and genome duplication is among the best-known examples of a recent allopolyploid speciation. Most Spartina naturally colonize intertidal zones of the American and
European Atlantic coasts, and the especially effective sand-fixer S. anglica has been widely introduced, often with massive colonization episodes and marshes silting.
We are developing genomic resources for Spartina in an effort to better understand the changes accompanying hybridization and polyploidy, focusing on S. anglica and its parents. 454 GS FLX pyrosequencing data were used to reconstruct the plastid sequence of the Euro-African hexaploid S. maritima , one of the parents. The 993,229 obtained reads were BLAST-searched against chloroplast genomes of other Poaceae, yielding 25,568 reads corresponding to plastid DNA. The assembled plastid genome was then annotated using Saccharum officinarum as a reference. Results confirm the conserved gene order of grass plastid genomes. A relaxed molecular clock (assuming uncorrelated rates) applied to coding
( rbc L, mat K, ndh F) and non-coding (introns and intergenic spacers) regions of 12 of the 13 species yielded a divergence time between the tetraploid and hexaploid clade of <5 million years (Ma). Because of the uniparental inheritance of plastid genomes, these estimates depend on depends on whether the hexaploid species ( S. maritima and S. alterniflora ) inherited their plastids from their tetraploid ancestor or from their diploid ancestor. Our best estimate is that S. maritima and the American S. alterniflora the parents of S. anglica , diverged around 3 my ago. These results provide a temporal framework for our ongoing work on the evolution of the transcriptomes of S. anglica and other allopolyploid Spartina .
The functional role of intervessel and intertracheid pit structure on cavitation resistance in angiosperm wood
Lens, F 1
E 5
, Sperry, JS 2
, Jansen, S 6
, Christman, M 3 , Rabaey, D 4 , Smets,
4
1 National Herbarium of The Netherlands, Leiden
University, Leiden, The Netherlands;
University of Utah, Utah, USA;
University of Utah, USA
2 Biology Dept,
3 Biology Dept,
Laboratory of Plant Systematics, K.U.Leuven, Leuven,
Belgium; 5 National Herbarium of The Netherlands,
6 Leiden University, Leiden, The Netherlands; Institute for Systematic Botany and Ecology, Ulm University,
Ulm, Germany
Fine-scale anatomical observations of the pit structure in
28 species belonging to several nonrelated lineages, including Acer, Artemisia, Ceanothus–Rhamnus, Lycium and Rosaceae, were carried out using the same native populations that were previously collected to perform hydraulic measurements. All species were observed using transmission electron microscopy to link ultrastructural pit characters with mean cavitation resistance (MCP). The 28 species were selected based on their differences in wood anatomy: seven contain a considerable number of tracheids that are associated near vessels (vasicentric tracheids in Ceanothus, Rhamnus and
Lycium ) and nine show true tracheids throughout the ground tissue (Rosaceae), while the other species studied
123

are characterized by non-conducting fibres. Preliminary observations show that intervessel pit membrane thickness is the most tightly correlated pit character with
MCP when all species are taken into account. Within specific clades, the pit characters correlated with MCP vary, as does the strength of individual correlations. This suggests that most pit correlations are clade-specific implying that resistance to cavitation can be achieved in many different ways. For instance, Ap (= mean pit area per vessel) is most tightly correlated with MCP in
Rosaceae, but not in the other clades studied. Likewise, the strong correlation between pit chamber depth and
MCP in Acer is weak or absent in the other clades. An exception to this clade-specificity is pit aperture fraction
(= pit aperture area per pit membrane area), which is linked with MCP in all but one clade. Among the species having vessels as well as true or vasicentric tracheids, the intervessel pit characters generally show higher tradeoffs with MCP than the intertracheid pit features. When vessel and tracheid pits are compared at the species level, however, there is a general trend for intertracheid pit membranes to be thicker than vessel-tracheid pit membranes, which are in turn thicker than intervessel pit membranes. These preliminary observations support the idea that tracheids act as a safe subsidiary transport mechanism in case too many vessels embolise due to prolonged drought stress.
Formation of freeze/thaw-induced embolism
Ball, M 1
1 Australian National University, Canberra, Australia
While much progress has been made on understanding the mechanisms underlying formation of freeze/thawinduced embolism, many questions remain. Cryo-
Scanning Electron Microscopy (CSEM) was combined with conventional hydraulic techniques to explore the occurrence of freeze/thaw induced embolism in the snow gum, Eucalyptus pauciflora . Measurements were made on detached leaves, detached stems with leaves, and intact plants, and compared with characteristics of samples collected from field-grown plants during natural frost events. Freezing typically initiated in the midvein near the petiole and spread throughout the leaf in approximately 10 s at -4˚C. Exotherms measured on attached, intact leaves show that temperatures remain elevated due to latent heat released during freezing for over 30 min following nucleation events. Such prolonged heating requires movement of water to sites of freezing and raises the possibility that the freezing of leaves may withdraw water from unfrozen stems. Detached leaves supplied with water showed a rapid increase in uptake once freezing began, with average uptake rates of 20 mg per hour sustained with cooling of leaves at 2˚C per hour to -8˚C, declining sharply to undetectable levels with further cooling from -10 to -14˚C. Using dyes to follow the path of water, we found that the uptake of water into freezing leaves occurred via unfrozen vessels. The decline in uptake as freezing progressed was associated with decrease in the number and size of unfrozen vessels.
When flow through stems was blocked, uptake of water by freezing leaves induced cavitation of stem xylem.
Similar measurements were made on intact plants that were either well watered or drought stressed. Drought stress induced loss of stem hydraulic conductance due to
124 embolism. During freezing of these droughted plants, however, the embolized vessels filled with ice as liquid water was drawn into the vessels through their walls.
Vessels remained filled with water and hydraulic conductance was restored following thawing from a moderate freezing temperature of -6˚C, whereas the fraction of embolized vessels was greater after thawing from -12˚C, apparently due to greater tension exerted by greater tissue dehydration at the lower nadir temperature.
The results show that vessels freeze independently and that the probability of freezing depends on vessel diameter and the extent of supercooling, with far reaching consequences for understanding the importance of freeze-induced redistribution of water within whole plants to understanding freeze/thaw-induced embolism.
Aquaporins and control of water flow
Tyerman, SD 1
MC 2 , Dayod, M
Kaiser, BN 1
,
1
Vandeleur, RK 1 , Bramley, H
, Tataranni, G 4 , Gilliham, M 1
2 , Shelden,
, Mayo, G 5 ,
1 School of Agriculture Food and Wine, University of
Adelaide, Waite Campus, Australia;
Agriculture, The University of Western Australia,
Crawley, Australia; 3
2 Institute of
Australian Centre for Plant
Functional Genomics, School of Botany, University of
Melbourne, Parkville, Australia; 4 Dipartimento Di
Scienze Dei Sistemi Colturali, Forestali E Dell'Ambiente
Università Degli Studi Della Basilicata Viale Dell'
Ateneo Lucano, Potenza, Italy; 5 Australian Centre for
Plant Functional Genomics, School of Agriculture, Food, and Wine, University of Adelaide, Waite Campus,
Australia
In the biosphere plants may conduct huge amounts of water through aquaporins, but there are unanswered questions about how aquaporins work and how they link to other nutrient transport such as nitrogen and Ca 2+ .
There are a variety of mechanisms controlling aquaporin activity in plant membranes, notably cytosolic pH and
Ca 2+ concentration. The effect of Ca 2+ is additionally interesting because its transport in the plant is largely coupled to water flow. These enable rapid responses to sudden environmental changes. Abiotic stresses such as drought and hypoxia have interesting effects on aquaporin mediated water transport that differ between species and cultivars within a species. Root anatomy and morphology are linked with the role of aquaporins in these abiotic stresses. The differences between cultivars in their water transport physiology present an opportunity to correlate the physiology with differences in gene expression. Root hydraulic conductance, which is linked with the expression of a PIP aquaporin in grapevine, is correlated with leaf transpiration indicating that long distance signalling is occurring. This can be observed in a variety of species as a correlation between root hydraulic conductance and leaf conductance or transpiration. This ultimately allows regulation to achieve maximum extraction of water from the soil.
Understanding these signals will be as important as is our understanding of the long distance signalling involved in stomatal regulation.

Biology of xylem refilling
Zwieniecki, M 1 , Secchi, F 1
1 Arnold Arboretum of Harvard University, USA
The process of xylem refilling requires coordinated action of stem parenchyma cells adjacent to the embolized vessels. Such coordination must rely on the plant ability to detect embolism which is temporally and spatially localized. We suggest that continuous transpiration in functional vessels removes signal molecules (sugars) released from parenchyma cells preventing their accumulation in conduit walls. If transpirational stream ceases following cavitation, signaling molecules will accumulate in the xylem walls allowing for localized detection of embolism. This detection has to be followed by a biological response that provides energy and facilitates water transport to the site of refilling. Here we present an overview of the Populus trichocarpa transcriptome response to embolism formation. This response includes changes in the expression pattern of genes belonging to diverse gene families including aquaporins, sugar transporters and sugar metabolic pathways. We discuss the transcriptome activity for its consistency with formerly proposed models of xylem refilling process.
Embolism resistance and refilling in plants: new insights using novel imaging techniques
Choat, B 1 , McElrone, A 2 , Brodersen, C 3
1 The Australian National University, USA; 2
Dept of Agriculture, USA; 3
United States
University of California,
Santa Cruz, USA
Plants are capable of rapidly transporting water to heights in excess of 100 m, and of extracting water from extremely dry or saline substrates. To achieve this, plants have evolved a transport system that relies on water sustaining a tensile force, such that the xylem sap is at negative absolute pressures. However, this transport mechanism comes with its own set of problems, most notably that water under tension is prone to cavitation, which results in the formation of a gas bubble
(embolism). Embolism reduces the capacity of the xylem tissue to deliver water to sites of gas exchange and can therefore impact the ability of the plant to maintain a net positive carbon balance. In the extreme, xylem embolism can reach lethal levels causing branch die back and ultimately plant death. Resistance to embolism is now recognised as a key trait determining the extent of woody plant mortality during drought and the limits of plant distribution with regards to water availability. Hydraulic constraints on plant growth and survival also clearly play a role in declining productivity in natural and agricultural systems during prolonged and severe droughts. Plants can avoid extensive build up of embolism in two ways.
First, by refilling conduits after they have cavitated, and second, by achieving hydraulic design that reduces the risk of embolism spreading throughout the xylem as water stress increases. The basic principles of water transport in plants are regarded as well known and there is a general consensus that the cohesion-tension mechanism is the driving force of water movement through the xylem. However, discoveries in the last two decades demonstrate that there are still fundamental gaps in our knowledge of long distance water transport in plants. I will address two outstanding questions to how plants transport water, (a) the ability to plants to refill embolised conduits during active transpiration and, (b) the factors that control the relative resistance to drought induced embolism in plants. Recent insights into embolism formation and refilling gained by x-ray microtomography and magnetic resonance imaging will be presented in the context of structure-function relationships.
Nighttime transpiration and xylem embolism repair in plants: two mysteries explain each other
Schenk, HJ 1
CS 2
, Espino, S 1 , Martínez-Cabrera, H 2 , Jones,
1 California State University Fullerton, USA; 2 of Connecticut, USA
University
Two enduring mysteries of plant physiology are the function of nighttime transpiration and the nature of the mechanism that allows removal of xylem embolisms while a plant’s hydraulic system is functioning under negative pressure. Nighttime transpiration is particularly common in plants from dry environments, which would appear to be least able to afford wasteful water loss.
Refilling of air-filled conduits while the remaining hydraulic system is under negative pressure has been thought to be physically impossible, yet is commonly observed. Here it is shown that the two mysteries can explain each other. The North American desert shrub
Encelia farinosa (Asteraceae) was used as the study system for this research. In this species embolism repair under negative pressure occurs usually at night while the stomata are open and while water potentials are below those potentially generated by root pressure.
Experimental inhibition of nighttime transpiration by bagging of leaves inhibits embolism repair. These findings show that, at least in this species, a transpiration stream is required for embolism repair under negative pressure. Measurements of air flow into artificially created embolisms in the wood revealed that air from these embolisms dissolves into the transpiration stream of functioning conduits. It is hypothesized that air from refilling xylem conduits diffuses through pit membranes into the transpiration stream of functioning conduits.
This happens while temperatures decline during the night, causing gas solubility in xylem sap to increase.
Nighttime transpiration appears to be required to move air-saturated sap towards the leaves before temperatures increase again during the next morning, which would cause gas solubility to decrease and air to come out of solution. Thus xylem appears to operate as a liquid-gas membrane contactor, which is an engineered device that efficiently moves gas via diffusion through large gaspermeable membranes that separate a gas and a liquid phase. Measurements of dissolved gas in xylem sap in woody roots and stems of Encelia farinosa fully supported this theory. The findings should help to put to rest enduring criticisms of the tension cohesion theory of water transport, which are largely based on the false assumption that negative-pressure hydraulic systems cannot self-repair after suffering embolism formation.
Nighttime transpiration, an important flux in the water balance of many ecosystems, is explained at least in part
125

as playing a vital role in the nocturnal recovery from drought-stress experienced during the day.
Are hygroscopic particles on the leaf surface part of the plant hydraulic system?
Burkhardt, J 1 , Hunsche, M 1 , Pariyar, S 1
1 University of Bonn, Germany
The hydraulic system of plants is generally assumed to end near the leaf surface but always still within the leaf.
It is assumed that all liquid water will evaporate there and that only water vapor will exit the plant via the stomata or the cuticle. The establishment of thin liquid water connections along stomatal walls might extend the hydraulic system to the leaf surface and intensify its coupling to the atmosphere, which would support earlier studies pointing to high frequency coupling between atmospheric turbulence and sap flow. In this contribution, we investigate how this 'hydraulic activation of stomata' (HAS) may establish, using environmental scanning electron microscopy to observe drying/wetting cycles of a range of different salts on hydrophobic leaf cuticles. We also investigate the response of stomatal conductance and transpiration to changes in relative humidity. The water vapor absorption to hygroscopic salts is a direct function of relative humidity, and together with HAS this is able to explain the stomatal humidity reaction. This includes the
'feedforward' effect (i.e. decreasing transpiration caused by decreasing relative humidity), which is interpreted as an emergent reaction of a stomatal patch. For this part of the study we use plants grown in particle free environment, in comparison with plants from normal air, and plants treated with different salt solutions. Different salts, placed on hydrophobic tomato cuticles, responded differently to humidity changes around their deliquescence point, but usually the salts expanded after several drying/wetting cycles, sometimes in dentritic form. This shows the capacity to overcome the hydrophobicity of leaf cuticles under conditions of high ionic strength, and further confirms the recently observed capability of thin water films to extend into the stomata.
The humidity reaction of stomata and plant transpiration differed between plants from the particle free environment and plants with different kinds and amounts of particles on their leaf surface. This supports the idea that leaf surface particles are an important part of plant/atmosphere interactions, and may eventually form the end of the plant hydraulic system. Coupling to the atmosphere is intensified by the degree of HAS and may be beneficial at moderate HAS, when acting as ‘humidity sensor’. Strong HAS (e.g. caused by air pollution) may affect plant water relations by wicking of liquid water out of the stomata, with hygroscopic leaf surface particles acting as desiccants.
Coordinated development of veins and stomata matches hydraulic supply with potential transpiration
Brodribb, T 1
Chang, Y 1
, Jordan, G 1 , Lam, V 1
, Brodribb, T 1
, Biffin, E 1
, Graham, SW 1
, Rai, H 1 ,
1 University of Tasmania, Hobart, Australia
Higher leaf vein density enables higher rates of photosynthesis because enhanced water transport allows higher leaf conductances to CO
2
and water. However the total cost of leaf venation rises in proportion to the density of minor veins, meaning that efficient investment in leaf xylem requires minor vein synthesis to match potential transpirational demand. Under conditions where maximum photosynthesis and stomatal conductance are limited by light intensity, optimal allocation of carbon for vein production should occur if variation in vein density
(Dv) is coordinated with variation in stomatal density.
Here we examined leaves of the evergreen tree
Nothofagus cunninghamii to determine whether this was the case. Sun and shade-grown leaves were compared across five populations ranging in altitude and rainfall, and a very strong correlation between Dv and stomatal density was found at all levels of comparison ranging from within-trees to among-populations. Calculated leaf conductances to liquid and vapour-phase water were strongly correlated over a two-fold range in Dv and stomatal density. Our results provide strong evidence that the development of veins and stomata are coordinated in such a way as to produce a highly efficient use of carbon investment in leaf venation. The developmental mechanisms that may integrate veins and stomata are discussed.
Hydraulic properties and photosynthesis in six deciduous and six evergreen tree species in a tropical limestone forest in southern Yunnan, China
Fu, P 1 , Jiang, Y 1 , Wang, A 1 , Cao, K 1
1 Key Laboratory of Tropical Forest Ecology,
Xishuangbanna Tropical Botanical Garden, Chinese
Academy of Sciences, Yunnan, China; 2 Graduate
University of Chinese Academy of Sciences, Beijing,
China
Trees growing on limestone will face frequent drought because of the low water retention capacity of the shallow soil, especially in the 6-month long dry seasons in SW China. And the evergreen and deciduous tree species growing in limestone may use different strategies to adapt to the seasonal drought. In the present study, we compared the stem hydraulic conductivity, leaf gas exchange rates, leaf pressure volume (P-V) curves traits, and leaf water status of six evergreen and deciduous tree species both in dry and wet seasons in a tropical limestone forest in Southern Yunnan, China. We also compared stem vulnerability curves and xylem anatomy of the 12 tree species. Our results showed that the leaf specific hydraulic conductivity, sapwood specific conductivity (K
S
), and maximum leaf mass based net photosynthesis rate (A m
) were much higher in the six deciduous tree species than those of the six evergreen tree species. And the deciduous tree species had much wider vessel diameter than that of the evergreen tree species. The evergreen tree species had lower xylem water potential at 50% loss of hydraulic conductivity than that of the deciduous tree species. The difference of
K
S
between the wet and dry season were not significantly different in most of the 12 species. However the K
S
of one deciduous species Bauhinia variegata (L.) in the dry season was significantly lower than that of the wet season, which might be caused by xylem embolism; and
126

the K
S
of another deciduous tree species Millettia cubitti
(Dunn) in the dry season was higher than that in the wet season, which might be due to the newly growing sapwood in the dry season. The turgor loss point (TLP) that calculated from the P-V curves for leaves or terminal shoots of the evergreen species were more negative than those of the deciduous tree species in dry season, which means the evergreen tree species have greater ability to maintain their leaf turgor than the deciduous tree species.
Our results indicate that there is a trade-off between hydraulic efficiency and safety: the evergreen species exhibit greater ability of drought-resistant with lower P
50 while the deciduous tree species show greater water transporting efficiency and photosynthesis rate in the rainy season. The seasonal exchange of K
S
shows that the declining of hydraulic conductivity of the stem may not be responsible for the leaf shedding of the deciduous tree species.
Linkages among diel carbon and water fluxes in six
Eucalyptus species
Lewis, JD 1, 2 , Phillips, NG 3 , Logan, BA 4 , Tissue, DT 2
1 Fordham University, USA;
Sydney, Australia; 3
2 University of Western
Boston University, USA; 4 Bowdoin
College, USA
Nocturnal water loss via transpiration is substantial in many plant species. While nocturnal transpiration may increase total daily carbohydrate production by increasing early-morning net photosynthetic rates and nutrient uptake, it may also reduce production by increasing water stress. However, the relationships between diel carbon and water fluxes in leaves are poorly understood. In this study, we examined leaf-level carbon and water fluxes of six Eucalyptus species, differing in drought sensitivity, during two 24-hour cycles in a common garden experiment at the University of Western
Sydney in Richmond, NSW. Drought-sensitive species generally exhibited lower net photosynthetic rates (A) and stomatal conductances (gs) at sunrise and midday, and A and gs values peaked earlier in the day than in drought-tolerant species. Variation in light-saturated net photosynthetic rates (Asat) reflected gs and starch accumulation during the day, and the relationship between Asat and gs did not vary among species.
Nocturnal respiration rates varied with leaf N concentration and the rate of soluble sugar loss at night.
As a result, there were no clear relationships between daytime and nocturnal carbon and water fluxes. These results suggest that drought-tolerant Eucalyptus species may maintain higher photosynthetic rates for longer periods of the day than drought-sensitive species.
Water-use of
Eucalyptus species originating from
1 differing climates: is canopy conductance modulated by species?
Bourne, A 1 , Haigh, A 1 , Ellsworth, D 1
Hawkesbury Institute for the Environment and School of
Natural Sciences, Australia
Transpiration by plants controls a major portion of water loss from land surfaces as mediated by stomata and the environmental conditions in which they function. There is currently limited information on species comparisons of transpirational water use for ecologically contrasting
Eucalyptus species despite the predominance of this genus on the Australian continent and in tropical and subtropical plantations world-wide. We asked whether there are differences in canopy conductance and its relationship to the environment among species adapted to contrasting climates. We investigated canopy scaled stomatal conductance (Gc) to determine how tree canopy conductance responded to climate variation amongst sub humid (SH) and humid (H) zone Eucalyptus species across a year. Measurements were made on four tree species from contrasting climate zones, namely E. crebra
(SH), E. dunnii (H), E.
melliodora (SH), E. saligna (H), and E. tereticornis (H). Sap flow techniques were used for the measurements, which were then scaled to canopy conductance by inverting the Penman Monteith formula.
Species originally from contrasting climate zones had different maximal Gc values, as well as different Gc responses to vapour pressure deficit (D). From different maximal Gc's at low D, different species converged on the same minimum Gc at high D. Sub humid Eucalyptus species showed a shallower response of Gc to D than did humid zone Eucalyptus species. This response to increasing D meant that sub humid species used larger amounts of water during dry and hot conditions, whereas humid zone species used larger amounts of water during humid and hot conditions, corresponding to the prevailing conditions in their climates of origin. Thus differences in Gc and the total water use response to changing environmental conditions were attributed to the original climate of the species. Understanding how
Eucalyptus water-use is regulated by stomata and how native species differ in this regard is critical for managing water resources.
4
The evolution of C
4
photosynthesis in the eudicots: lessons for understanding C
4 other monocots
evolution in grasses and
Sage, R 1 , Sage, T 1
1 University of Toronto, Canada
There are currently 62 lineages of C
4
photosynthesis in the plant kingdom, of which 36 are in the eudicots and 18 are in the Poaceae. The C
4
grass lineages have proven
origin, because difficult for addressing hypothesis of C
4 their taxonomy and phylogenetics are currently in flux, many lineages lack close C
C
3
C
–C
4
3
relatives, and there are few
intermediate species. A major exception is the small Australian genus Neurachne , which has numerous
species and a C
3
–C
4 3
species, one C
4
intermediate species. While the anatomy and physiology of Neurachne are well studied, its phylogenetics remain vague, limiting its value for evolutionary work. By contrast, the lineages of C
C
3
4
dicots have been valuable for addressing hypothesis regarding C
–C
4
4
evolution. Multiple species level phylogenies exist in the C
4
clades, there are numerous
intermediates, and closely related C
3
and C species have been identified. Thus, early work on C
4
4 evolution in Flaveria has been built upon by comparative
127

studies of eudicot genera such as Heliotropium, Cleome,
Chamaesyce, Portulaca and Mollugo . These studies demonstrate conclusively that C
4 via the evolution of a photorespiratory CO
2 mechanism, which appeared in C
3
photosynthesis arose
concentrating
species of very hot, monsoon-affected climates where photorespiration is high. C
3
species with high vein density and enlarged bundle sheath cells appear predisposed to evolve C
4 photosynthesis. Our working hypothesis is that high vein density in C
4
ancestors occurred in order to transport water to the mesophyll cells in hot, low humidity environments where transpiration is extreme. With high vein density there are often high numbers of bundle sheath (BS) cells, which are fully incorporated into the leaf photosynthetic apparatus by an increase in chloroplast number along the BS periphery. To partially offset high photorespiration, it is possible that a photorespiratory loop is established along the length of the BS cells, with photorespiratory metabolites diffusing from the outer BS periphery to the inner cell region where mitochondria and glycine decarboxylase (GDC) are localized. Here, release of photorespired CO
2
can be efficiently refixed by adjacent chloroplasts. This possibility is supported by close studies of the immediate
C
3
relatives of the C
4
lineages in Heliotropium and
Flaveria , where mitochondria and glycine decarboxylase only occur along the inner BS wall. The establishment of a BS photorespiratory loop could enable the subsequent establishment of a mesophyll to BS photorespiratory cycle via a mutation that eliminates mesophyll GDC expression. By establishing high numbers of photosynthetic BS cells, and localizing the BS mitochondria to the inner wall, C
3
species are predisposed to survive a loss of mesophyll GDC, thereby allowing evolutionary optimization of an efficient photorespiratory CO
2
concentrating mechanism.
The evolution of beta carbonic anhydrases in C
4 plants
1
Ludwig, Martha 1
University of Western Australia, Crawley, Australia
Both C
3
and C
4
plants contain multiple isoforms of the enzyme
β
-carbonic anhydrase (CA), which catalyse the interconversion of CO
2
and bicarbonate. In C
3
plants, most CA activity localizes to the chloroplasts of the mesophyll cells. C
3
plastidial
β
-CAs have diverse roles, including the facilitation of CO
2
diffusion across the chloroplast envelope, lipid biosynthesis, disease resistance, and stomatal closure. By contrast, high levels of CA activity localize to the cytosol of C mesophyll
4 cells where it catalyzes the hydration of atmospheric
CO
2
, producing bicarbonate, the substrate for phosphoenolpyruvate carboxylase (PEPC), the primary carboxylating enzyme of C
4
plants. Most of our current understanding of the molecular changes responsible for the differences in CA expression patterns between C
C
4
3
and
plants has been gained through examination of the
CAs in species within the dicot genus Flaveria . This group of plants contains species that do either C
3
NADP-malic enzyme-type C
4 other species that carry out C
or
photosynthesis, as well as
3
–C
4
intermediate types of photosynthesis. Results from immunocytochemistry, chloroplast import studies, quantitative reverse transcription polymerase chain reaction assays and
128 sequence analyses indicate that the gene coding for an ancestral C
3
chloroplastic CA was the evolutionary template for the C
4
cytosolic form of the enzyme in
Flaveria , with the loss of the sequence encoding the chloroplast transit peptide from the C
3
CA gene leading to the retention and operation of the C
4
CA in the mesophyll cytosol. Analysis of the upstream region of the C
4
CA gene identified a sequence similar to that of the mesophyll expression module 1 (Mem1) that directs mesophyll cell-specific expression of the PEPC gene in
C
4
Flaveria species. When this CA gene upstream region was used in a promoter/reporter gene construct to transform F. bidentis plants, reporter gene expression was detected specifically in the mesophyll cells of this C
4 species. With respect to the Flaveria intermediate species, cDNA sequence analyses indicated that the C
4
like species F. brownii expresses an ancestral chloroplastic C
3
C
4
-like species,
form of the CA gene while in another
F. vaginata , a cytosolic C
4
form of the
CA3 gene is transcribed. These results indicate that different biochemistries and/or gene expression patterns can result in similar photosynthetic physiologies, and that different molecular evolutionary paths may have been taken to achieve C
4
-ness in Flaveria . Recent work on the molecular evolution of the
β
-CAs in Cleome , another dicot genus that contains C type C
4
3
and NAD-malic enzyme-
species, as well as insights into the molecular evolution of
β
-CA gained from studies focusing on grasses, will also be discussed.
Efficiency of the CO
2 during C
4
concentrating mechanism
photosynthesis: response to changes in light availability and potential of the single-cell C
4
, King, J 1 , Ubierna Lopez, N 1 ,
plants
Sun, W 1
1
Cousins, A 1
Washington State University, USA
Of the total solar energy reaching the Earth’s surface, only a portion is used by higher plants to convert inorganic carbon to organic carbon. This is in part because under current atmospheric CO photorespiration in C
3
2
concentrations,
plants decreases the theoretical maximal efficiency of converting solar energy into chemical energy. In response, some species have evolved
C photosynthesis, which reduces photorespiration rates
4 by capturing atmospheric CO
2
in the C
4
cycle and concentrating it around the compartmentalized Rubisco and the C
3
cycle. The efficiency of the C
4
CO
2
-concentrating mechanism is defined in part by the fraction of CO
2
fixed by the C
4
-cycle that subsequently leaks out of the compartmentalized C
3
photosynthetic
-cycle and is not assimilated by Rubisco. Both the relative biochemical capacities of the C
CO
2
from the compartmentalized Rubisco determine the amount of CO
2
4
and C
3
‘leakiness’. The use of leaf-level models and measurements of 13 CO
cycles and the conductance of
2
isotope discrimination (
δ
13 are powerful tools to estimate the efficiency of the CO
C)
2
concentrating mechanism (Farquhar, 1983). The goal of our research is to use modeled and measured values of
δ
13 C to determine how the efficiency of the CO concentrating mechanism during C
2
-
4
photosynthesis is influenced by changes in light quantity and quality, and by variations in anatomical structures used to facilitate C
4 photosynthesis. Light availability, quantity and quality, are expected to impact the coordination of the C
4
and C
3 cycles, especially if there is unequal distribution and

absorption of light energy within a leaf. Indeed, the rate of C
4
photosynthesis decreases and CO
2
leakiness appears to increase in response to low light availability.
Additionally, CO
2
leakiness changes when illumination shifts from red to blue and blue to red light. Data will be presented describing the response of leakiness and potential mechanisms influencing leakiness in response to changes in light quantity and quality. In addition to the coordination of the C
CO
2
4
and C
3
cycle, the conductance of
diffusion between the compartmentalized C cycles is required for efficient C
4
4
and C
3
photosynthesis. To maintain a low conductance most terrestrial C
4
plants utilize two distinct photosynthetic cells (Kranz anatomy) to concentrate CO
2
around Rubisco and minimize rates of photorespiration. However, the Chenopodiaceae species
Bienertia sinuspersici uses central and peripheral cytoplasmic compartments and Suaeda aralocaspica utilizes distal and proximal compartments, each within a single-cell, to facilitate C
4
photosynthesis. There is strong evidence that these plants perform C
4 photosynthesis; however, questions remain regarding the photosynthetic capacity and efficiency of these singlecell CO concentrating mechanisms. Data will be
2 presented demonstrating that the rates of photosynthesis
concentrating mechanisms and the efficiency of the CO
2 of the single-cell C aralocaspica are similar to those of the more common
Kranz-type C
4
plants.
4
plants B. sinuspersici and S.
Global environmental change and the future of C
4 crops for food and fuel
Leakey, ADB 1 , Markelz, RJC 1 , Strellner, RS 1
1 University of Illinois At Urbana–Champaign, USA
Crops with the C
4
photosynthetic pathway are vital to global food supply, particularly in the Americas and
Africa. While rising atmospheric [CO
2
] is the driving force behind the greater temperatures and water stress which threaten to reduce future crop yields, it also has the potential to directly benefit crop physiology. The nature of C
4
plant responses to global environmental change has been controversial. Recent evidence from
Free-Air Concentration Enrichment experiments suggests that elevated [CO
2
] does not directly stimulate C
4 photosynthesis. Nonetheless, drought stress can be
] as a result of lower ameliorated at elevated [CO
2 stomatal conductance and greater intercellular [CO
2
].
Therefore, unlike C
3
crops for which there is a direct enhancement of photosynthesis by elevated [CO crops will only benefit from elevated [CO
2
2
], C
4
] in times and places of drought stress. This talk reviews the results from recent field experiments investigating the response of maize to the CO
2
x drought interaction, as well as a new project aiming to resolve a key remaining knowledge gap - whether rising temperatures will directly sensitize C
4
photosynthesis to elevated [CO
2
].
1
Ecophysiology of C
4
grasses: a comparative approach
Osborne, C 1 , Taylor, S 2 , Ripley, B 3 , Woodward, I 1
University of Sheffield, Sheffield, UK;
Texas, Austin, USA;
South Africa
3
2 University of
Rhodes University, Grahamstown,
C
4
photosynthesis ranks among the most important evolutionary innovations in plants, and the assembly of ecosystems dominated by C
4
grasses has transformed the terrestrial biosphere. The rapid accumulation of phylogenetic evidence over the past decade has revealed multiple evolutionary origins of the C
4
pathway spanning a diverse range of grass lineages. This accumulated knowledge brings new opportunities for understanding the functional significance of the C syndrome within its
4 evolutionary context. In this talk, we will consider the question: what changes in plant water relations and nitrogen-use have been associated with the evolution of
C
4
photosynthesis in grasses? First, a comparative screening experiment used a total of 34 grass species to sample five independent C
4 groups of C
3
origins, including three
sister taxa. Data supported the a priori expectations of lower stomatal conductance, and greater water- and nitrogen-use efficiency in C
4
than C
3
species.
Further data showed that the soil-leaf water potential gradient was greater in C
3
than C
4
species. However, differences in photosynthetic rate and leaf nitrogen concentration were smaller than expected. A second comparative screening experiment compared the physiological responses to drought in a phylogenetically structured sample of 14 C seven independent C
4 the C
4
3
and C
4
grass species, covering
origins. Under mesic conditions,
grasses had higher photosynthetic rates, but lower stomatal conductance, and a smaller soil-leaf water potential gradient than their C
3
counterparts. Under drought, stomatal conductance declined more dramatically in the C
3
than C
4
species, and photosynthetic water-use and nitrogen-use efficiency advantages held by C
4
species under control conditions were each diminished by 40%. Again, there was only a small effect of photosynthetic pathway on leaf nitrogen concentration. Accounting for the phylogenetic diversity of C
4
grass species has therefore provided new insights into the evolutionary links between photosynthetic pathway and leaf water- and nitrogen-use. A modelling framework for drought mortality is used to interpret these results in the context of ecological selection for C
4 photosynthesis.
A phylogenetic perspective on the responses of C
3
C
4
grasses to fire
and
, Martin, T 1 , Osborne, C 2
1
Ripley, B 1
Botany Dept, Rhodes University, South Africa; 2 Dept of
Animal and Plant Sciences, University of Sheffield, UK
C
4
grasses possess characteristics that potentially advantage growth in fire-prone environments, including high photosynthetic productivity, efficient light and nutrient use and significant allocation to below-ground reserves. Such characteristics allow fast regeneration after fire, and may be the consequence of photosynthetic physiology, phylogenetic ancestry or may have been acquired as adaptations to frequently burnt environments.
The aim of this study was to examine the role of photosynthetic pathway by comparing the fire ecology of
14 species of C
3
and C
4
grasses belonging to the subfamilies Panicoideae (Panicoid), Aristidoideae and
Danthonioideae (non-Panicoid), and drawn from the same regional flora in the Eastern Cape of South Africa.
Its focus was on aboveground characteristics that would contribute to a fire fuel load and the re-growth responses
129

of plants subsequent to a controlled experimental burn during the natural winter fire season. Prior to the burn, but after frost, a greater proportion of the aboveground biomass of C moisture content, and higher flammability than that of the C
3
4
species was dead, and had a lower
plants. These characteristics would all potentially contribute to a greater fuel load and meant that C
3
plants lost a larger proportion of living tissue in the experimental burn. However, these patterns were equally strongly determined by phylogeny, and leaf mortality was greater in the Panicoid than the non-Panicoid grasses. Similarly, re-growth subsequent to burning was more rapid and complete relative to controls in Panicoid than non-Panicoid grasses, and did not differ between C
3 and C
4
grasses within each of these phylogenetic groups.
The re-growth of Panicoid grasses was supported by a stronger re-allocation of larger belowground reserves than was apparent for non-Panicoid grasses and again was not different between photosynthetic types. Results suggest that Panicoid grasses are tolerant of fire and would explain why the number of species is positively correlated to fire frequency. What is not evident is why
C
4
Panicoid grasses dominate frequently burnt ecosystems, as their performance after fire was not significantly superior to that of C
3
Panicoid grasses.
The biophysics of the rhizosphere
Young, IM 1
1 School of Environmental & Rural Science, University of
New England, Armidale, NSW, Australia
Much is know about life in and near the rhizosphere.
Relatively little is known of the consequences of this life in the context of the dark opaque and heterogeneous geohabitats of the rhizosphere, which is their home, as opposed to simple agar blocks. This talk will focus on these heterogeneous habitats and how the soil-microbe interactions within the rhizosphere are defined in many ways by the physical micro-architecture of the soil, and how in turn microbes define their habitats. Using high resolution micro-tomography the talk will explore the life of microbes in soil and what makes them what they are.
Bioactive root exudates: a novel source of allelochemicals and plant protectants
Weston, L 1
1 Charles Sturt University, Wagga Wagga, Australia
Plant root exudates are known to play an important role in community structure and are involved in complex rhizospheric interactions. Our past work with Sorghum spp. has elucidated the role of the allelochemical sorgoleone, a potent inhibitor of plant growth that is released in sorghum root exudates. Graminaecous species including fine fescue ( Festuca rubra ) also produce large amounts of novel secondary products and can selectively inhibit weeds in both field and laboratory conditions.
These constituents are known to play important roles in plant defense against herbivores, insects, pathogens and
130 microbes as well as competing plants. In Australia,
Paterson's curse ( Echium plantagineum ) also produces unique root exudates from two types of root hairs, which are involved in active exudation processes in the plant.
We have discovered that the periderm of both its younger lateral roots and older taproots produce unusual, redcolored chemical constituents which are localized in the outer layers of cells. We have determined the structure of these novel anthroquinones and discovered that they are potent inhibitors of certain microbes and pathogens.
Further analyses on their localization, mode of action, and biosynthesis are underway.
Measurement of allelochemical dynamics in the rhizosphere
Weidenhamer, J 1 , Mohney, B 1
1 Ashland University, USA
Plants produce a wide variety of highly phytotoxic chemicals, some of which have activities comparable to synthetic herbicides. The possibility that plants exert direct chemical, or allelopathic, effects on neighboring plants as well as on soil microflora has attracted considerable research interest. The success of certain invasive plants has been attributed to phytotoxic root exudates. However, the difficulties of monitoring allelochemical concentrations in soil and their dynamics over time have been a major barrier to testing hypotheses of allelopathic effects. We have used diffusive sampling strategies employing polydimethylsiloxane (PDMS) microtubing to map the spatial distribution and temporal dynamics of root-exuded thiophenes from the African marigold ( Tagetes erecta ) and French marigold ( Tagetes patula ). Solid phase root zone extraction (SPRE) probes were constructed by inserting stainless steel wire into
PDMS tubing. Alternately, 1 m lengths of PDMS microtubing have been placed in marigold soil with the two ends of the tubing remaining out of the soil so that solvent could be washed through the tubing to collect samples for HPLC analysis. This silicone tube microextraction (STME) method gave a linear response for á-terthienyl when maintained in soils spiked with 0-
10 ppm of this thiophene. These PDMS-based techniques are experimentally simple and use inexpensive materials, and should be broadly applicable to the measurement of non-polar root exudates. By allowing repeated sampling with minimal disturbance of the soil, these methods provide a means to test hypotheses about the role of root exudates in plant–plant and other interactions.
The evolution and functioning of fungal symbiosis
1 from liverworts to higher plants
Field, K 1 , Tille, S 1 , Cameron, D
University of Sheffield, UK
1 , Leake, J 1 , Beerling, D 1
It was first hypothesised over 30 years ago that symbiotic soil fungi assisted plants in ‘greening the Earth’ over 400 million years ago, by forming mutualistic arbuscular
‘mycorrhizal associations’ through which the fungus gains photosynthate in return for supplying host plants with mineral nutrients and water (Pirozynski & Malloch
1975). Crucial new evidence revealed that liverworts not only occupy the most basal node on the land plant

phylogenetic tree, but were probably the first plants to establish symbiotic partnerships with arbuscular mycorrhizal fungi (AMF) upon land colonisation (Wang et al. 2010). Essential genes required for mycorrhization in liverworts have been vertically inherited through to all the other major groups of land plants (Wang et al. 2010).
These advances in phylogenetics have exposed that virtually nothing is known about mycorrhizal functioning in lower land plants occupying major nodes on the land plant evolutionary tree. The remaining key to understanding the role of plant–AMF symbioses in the
‘greening of the Earth’ is now to resolve their functional relationships in lower plants. We measured carbon-formineral nutrient exchanges in plant–AMF associations in
Pressia quadrata , a thalloid liverwort, Osmunda regalis , a ‘lower’ tracheophyte fern and Plantago lanceolata , a modern ‘higher’ plant. Plants were grown at modern-day ambient (440 ppm) and under simulated Palaeozoic
(1500 ppm) atmospheric CO
2 concentrations.
Carbohydrate allocation from plant to AMF was determined by 14 CO
2
pulse-labelling of thallus or shoots and measurement of 14 C in external mycorrhizal mycelium growing into nylon mesh cores that exclude rhizoids and roots. Transfer of phosphorus absorbed by the fungi from within the cores into plant tissues was assessed by addition of 33P-labelled H
2
PO
4
tracer. The length of mycorrhizal mycelium in the soil supported by each plant provides a measure of surface area available for nutrient absorption and fungal biomass. In the liverwort and fern significant 14 C transfer from plant photosynthates to AMF was detected, but at rates that were more than two orders of magnitude lower than in the higher plant. Phosphorus uptake via the fungal partner showed a similar pattern of greater transfer in the higher plant than the lower plants. However, carbon cost per unit phosphous gained from the mycorrhizal symbiosis was lowest in Pressia and rose progressively through Osmunda to Plantago . This decrease in efficiency was in linear proportion to the hyphal lengths that increased from the liverwort through to the higher plant. Although net P uptake increased dramatically under elevated CO
2
, the carbon costs rose even more sharply resulting in loss of efficiency of the symbiosis compared to ambient CO , and this corresponded with
2 greatly increased hyphal lengths. Our results establish that AMF associations in lower land plants can function in the same mutualistic manner as in higher plants, both in early Palaeozoic and current CO
2
atmospheres. The carbon cost of AMF in the ‘lower’ plants is less than in the higher plants and has decreased with decreasing atmospheric CO concentrations. These findings provide
2 important new insights into the evolution and functioning of fungal symbiosis from liverworts to higher plants.
Competition for N in the rhizosphere of European beech forests
Simon, J 1 , Rennenberg, H 1
1 University of Freiburg, Institute of Forest Botany and
Tree Physiology, Germany
Beech-dominated deciduous forests constitute the potential natural vegetation in Central Europe. Like other forests, they developed on marginal soils characterised by N limitation. Thus, different biota in these forests have to share and, therefore, compete for the N resources.
For example, plant and microbial N fluxes are closely linked in terrestrial ecosystems. Plants may affect microbial N turnover by root exudation, whereas microorganisms may influence plant N fluxes by mineralisation of decaying biomass. When N is limiting in the soil, plants and microorganisms may compete for the same inorganic or organic N sources. This competition may even be enhanced under the prognosticated climate changes (i.e. elevated temperature, enhanced frequency and duration of summer droughts), particularly in tree species susceptible to strong and prolonged periods of drought. Such species
(e.g. European beech) may be negatively affected in their physiological performance and growth. To optimize the exploitation of limited soil N resources, trees have developed several mechanisms. However, the complex interactions between different components of vegetation, mycorrhizal fungi, and soil microorganisms in the competition for N and the regulation of these interactions are currently far from being understood. Therefore, the main objectives of this study were (1) to characterize and quantify the significance of vegetation components and soil microorganisms in the competition for N in Nlimited beech-dominated deciduous forests, and (2) to analyse the factors regulating this competition.
Molecular dissection of local and systemic responses
3
1 to phosphate starvation in
Arabidopsis thaliana
Nussaume, L
Arrighi, J-F 2
Nakanishi, T 6
1 , Desnos, T 1
, Bayle, V 3
, Clement, M 1 , Thibaud, MC 1
, Paz Ares, X 4 , Kanno, S 5
LBDP,UMR 6191 CNRS-CEA, Aix-Marseille II, CEN
Cadarach, St Paul lez Durance,France;
441/2594 INRA,CNRS, Castanet-Tolosan, France;
LRDP, ENS Lyon, France; 4
Cantoblanco, Madrid, Spain;
2
,
,
LIPM UMR
CNB, CSIC,Campus de
5 Graduate School of
Agricultural and Life Sciences,University of Tokyo,
Japan
Phosphate (p i
) is a crucial and often limiting nutrient for plant growth. Ìt is also a very insoluble ion heterogeneously distributed in soil. to cope with such a situation, plants have evolved a complex network of morphological and biochemical processes (1) controlled by various regulatory systems. these signalling pathways are triggered either by p medium (external p i i
concentration in the growth
; 2,3,4), or by plant cells internal P i
.
A split-root assay performed to mimic an heterogeneous environment combined with a transcriptomic analysis was used to identify clusters of genes locally or systemically regulated by P i
starvation (5). The combination of genetic tools and physiological analysis revealed distinct regulatory roles for the internal and the external P i
. They also pointed out a central role of the transcription factor phr1 for genes systemically controlled by low P i
. In addition, results obtained with a set of tools including chemical genetics and manipulations of the entire family of the high affinity P i transporters will be presented. These tools offer novel
131

powerful approaches to dissect P i
perception pathways.
(1) Misson et al. 2005. (2) Reymond et al. 2006. (3)
Svistoonoff et al. 2007. (4) Ticconi et al. 2009. (5)
Thibaud et al. 2010.
Translocation of miRNAs in phloem in response to phosphorus deficiency
Rodriguez-Medina, C 1 , Ho, A 2 , Loughlin, P 3 , Atkins,
CA 4 , Smith, PMC 3
1 Institut National de la Recherche Agronomique (INRA),
2 France; Dept of Molecular Biosciences, University of
Oslo, Norway; 3 School of Biological Sciences, The
University of Sydney, Australia; 4 School of Plant
Biology, The University of Western Australia, Australia
The phloem long-distance translocation system functions as a nutrient delivery system and as a signalling pathway through which molecules such as growth-regulators or bioactive peptides, proteins and RNAs are disseminated throughout the plant. This translocation of signals is involved in coordinating development and allowing the plant to respond to environmental conditions. However, the mechanisms controlling macromolecular trafficking and information flow are not well described. Small RNA molecules like small interfering RNAs (siRNAs), microRNAs (miRNAs) and noncoding RNAs that play an important role in the regulation of gene expression, have been detected in phloem exudate. We have identified 11 different miRNAs in L. albus phloem exudate through hybridisation and cloning. In
Arabidopsis we are investigating whether these miRNAs are translocated in response to nutrient stress using a grafting assay where wild-type tissue is grafted onto a hen1 mutant that is disrupted in miRNA biogenesis. The accumulation of miRNAs in scions and rootstocks was assessed by real-time quantitative PCR. In phosphorus deficient conditions miR399 was translocated as well as two other miRNAs known to respond to P deficiency in phloem. We are currently using small RNA sequencing to identify other miRNAs that are translocated. We are using a GFP sensor construct in Arabidopsis to study the process of translocation of miRNAs.
Phosphate signaling and homeostasis in plants
Pant, BD 1 , Scheible, WR 1
1 Max-Planck Institute of Molecular Plant Physiology,
Germany
Phosphate (P i
) is often limited in majority of the world soils for plant growth and development. To supplement the P i
-deficiency, phosphate is supplied to the soil as a fertilizer. The main source of phosphate fertilizer is phosphate rock which is a limited resource and will be finished in the near future. On the other hand, using excessive fertilizer is not eco-friendly. To cope with low
P i
availability, plants develop a number of adaptive features at morphological, physiological and molecular level. We have identified that a large number of genes are induced in response of P i
-deficiency using ATH1 array and real-time RT-PCR. Developing a real-time RT-
PCR based platform to quantify the primary transcript
(PTs) of all known Arabidopsis miRNAs, we identified many nutrient responsive miRNAs. Among those are
132 miR399s, miR827, miR778, miR156, miR169s, miR399* and miR778*. In addition, by small RNA library sequencing, we identified some additional Pistarvation inducible miRNAs including miR2111. miR399 is induced specifically during P goes down with P i
-resupply to P i i
-limitation and
-starved plants. miR399 negatively regulates an ubiquitin conjugating enzyme
(UBC24/PHO2) by transcript degradation as well as by translational inhibition. Therefore, overexpressor of miR399 (miR399OX) or loss-of-function PHO2 mutant
(pho2) leads to the same biochemical and molecular phenotype. miR399OX or pho2 leads to the accumulation of 2–4 fold high P i
in the shoot. Some P i
starvation inducible (PSI) genes including high affinity
Pi-transporter PHT1; 8 remain induced in miR399Ox or pho2 even under high P
P i
condition. This explains the high i
-accumulation phenotype of miR399OX or pho2 and shows that miR399/PHO2 module is involved in signaling P i
-limitation and allocation of P i
between shoot and root. We identified that PHR1 (a Myb-related transcription factor) is an upstream regulator of miR399.
PHR1 regulates a large number of PSI genes in addition to miR399. During P i
-deficient condition, different plant organs need to communicate and co-ordinate among each other. We identified that miR399 PT is induced to P i
limitation to a greater extent in shoot. The miR399 moves from shoot to the root via phloem stream and in down-regulates PHO2. Consequently, higher P supplied to the shoot to meet its P i i
is
-demand. To our knowledge, this is the first example of a miRNA shown to be a systemic signal. Recently, we have identified a number of new plant miRNAs in Arabidopsis and
Brassica by small RNA sequencing. We are characterizing some of these P reveal their role in P i i
-responsive miRNAs to
-signaling and homeostasis.
Furthermore, being a component of protein degradation pathway, the proteins that PHO2 will target for degradation are unknown and we are doing some experiments to find out its target proteins and its interacting partners.
Genetic and genomic evidence that sucrose is a global regulator of plant responses to phosphate starvation in
Arabidopsis
Lei, M 1 , Liu, Y 1 , Zhang, B
Wang, X-J 2 , Raghothama, KG 3
2 , Zhou, Y 2
, Liu, D 1
, Zhao, Y-T 2 ,
1 Protein Science Laboratory of The Ministry of
Education, School of Life Sciences, Tsinghua University,
2 Beijing, China; State Key Laboratory of Plant
Genomics, Institute of Genetics and Developmental
3
Biology, Chinese Academy of Sciences, Beijing, China;
Dept of Horticulture and Landscape Architecture,
Purdue University, West Lafayette, USA
Plants respond to phosphate (P i
) starvation by exhibiting a suite of developmental, biochemical, and physiological changes to cope with this nutritional stress. To understand the molecular mechanism underlying these responses, we isolated an Arabidopsis mutant, hps1
(hypersensitive to P i
starvation 1), which has enhanced sensitivity in almost all the aspects of plant responses to
P i
starvation. Molecular and genetic analyses indicated that the mutant phenotype is caused by overexpression of
SUCROSE TRANSPORTER 2 (SUC2) gene. As a consequence, the hps1 has a high level of sucrose in both

its shoot and roots tissues. Overexpression of the SUC2 or its closely related family members SUC1 and SUC5 in wild type plants recapitulate the phenotype of hps1. In contrast, the disruption of SUC2 functions greatly inhibits plant responses to P i
starvation. The microarray analysis further indicated that 73% of the genes that are induced by P i
starvation in wild type plants can be directly induced by elevated level of sucrose in hps1 mutants, even when they are grown under P i
sufficient condition. These genes include several important P i signaling components and those that are directly involved in P i
transport and mobilization. Interestingly sucrose and low P i
signals appear interact with each other both synergistically and antagonistically in regulating gene expression. Genetic and genomic studies provided strong evidence that sucrose is a global regulator of plant responses to P i
starvation. Keywords: P i
starvation response, hps1 mutant, sucrose, global regulator
Is the phosphate analogue phosphite interfering with plant phosphorus homeostasis and signalling?
Jost, R 1 , Pharmawati, M 2 , Berkowitz, O 3 , Pearse, SJ 1 ,
1
Lambers, H 1 , Finnegan, PM 1
School of Plant Biology, The University of Western
Australia, Crawley, WA, Australia; 2 Biology Dept,
Faculty of Mathematics and Natural Sciences, Udayana
University Campus Bukit, Jimbaran, Bali, Indonesia;
3 School of Biological Sciences and Biotechnology /
Centre for Phytophthora Science and Management,
Murdoch University, Murdoch, WA, Australia
Phosphorus (P) is a macronutrient that is essential for plant growth, but often has a low availability due to a low solution P pool and sorption to soil minerals. P is taken up by the roots of plants in the form of inorganic phosphate (P i
) and plants feature complex regulatory networks to maintain P homeostasis and optimise their P i uptake and storage capacities to meet metabolic and developmental demand. Phosphite (Phi, HPO32-) is a more reduced form of P that is used as a biostat to enhance plant resistance against Phytophthora species.
These pathogenic oomycetes are a major threat to both food security ( Phytophthora infestans caused the Irish potato famine in the mid 19th century) and natural diversity ( Phytophthora cinnamomi has been termed the
‘biological bulldozer’ due to its devastating effect on plant communities in many biodiversity hotspots around the globe). However, despite its successful marketing as a ‘fungicide’ relatively little is known about its longerterm impact on plant growth and development. Phi is believed to mimic P i
in suppressing the plant’s P i
starvation response and severely inhibits growth of plants with a low P status. Its uptake most likely proceeds via transporters of the PHT1 family and is highly sensitive to competitive inhibition by P i
. Here we show that Phi is not a perfect mimetic of P i
in suppressing P i
-starvation responses. While it does suppress root-hair formation and attenuate the expression of many P i
-starvation-induced genes, it does not reduce the expression of many other well known P i
-signalling genes. In contrast to P i resupply, Phi addition actually leads to a severe inhibition of primary and secondary root elongation and an increase in lateral root density that is much more pronounced than under P i
starvation itself. Anthocyanin accumulation in older leaves is not completely reversed by supplying Phi instead of P i
to P-starved plants. This could be an indication for altered shoot P i
reallocation patterns in those plants. We show that Phi evokes a set of distinct physiological and molecular reactions that distinguish it from well characterised P i
-induced changes and therefore make it an excellent tool to study P-sensing and -signal-transduction pathways. This will also provide new insights into how Phi alters plant defence responses to boost their resistance against parasitic oomycetes.
Tilling identifies mutations in key genes involved in the cyanogenesis pathway in sorghum
1
Blomstedt, C 1 , Neale, A
Stuart, P 3 , Hamill, J 1
1 , O'Donnell, N 1
, Gleadow, R 1
, Møller, B 2 ,
Monash University, Australia;
Copenhagen, Denmark; 3
2 University
Pacific Seeds, Qld, Australia of
Forage sorghum is an important pasture grass with high growth rates, providing good feed stock, and it is drought and heat tolerant. Thus, it is widely grown in dry, tropical regions worldwide. Sorghum plants produce a stable, non-toxic tyrosine-derived cyanogenic glycoside compound known as dhurrin. Dhurrin is a natural defence product that liberates prussic acid (HCN) when the leaf tissue is consumed. Young plants or those experiencing abiotic stresses, particularly drought, can be highly toxic, accumulating dhurrin to high levels. In
Australia alone there is loss of approximately $20 million a year from fields of sorghum that cannot be grazed, due to fears of toxic poisoning. We are using TILLING
(Targeted Induced Local Lesions in Genomes) to identify induced point mutations in a number of key genes in the cyanogenesis pathway of sorghum. These mutant lines are currently being characterised for growth rate, nitrogen use efficiency and stress tolerance. The aim of our research is to produce novel sorghum lines with reduced prussic acid levels.
Smoking out the masters: clustered transcription
1 factors regulating nicotine biosynthesis in tobacco
Shoji, T 1 , Kajikawa, M 1 , Hashimoto, T 1
Nara Institute of Science And Technology, Japan
Alkaloids are one of the most diverse families of natural products derived from plants; over 12,000 different chemical structures have been discovered and some of them exploited as important pharmaceutical agents.
Tobacco ( Nicotiana tabacum ) synthesizes nicotine and related pyridine alkaloids in the root, and their synthesis increases upon herbivory on the leaf via a jasmonatemediated signaling cascade. Regulatory NIC loci that positively regulate nicotine biosynthesis have been genetically identified, and their mutant alleles have been used to breed low-nicotine tobacco varieties. Here, we report that the NIC2 locus, originally called locus B, comprises clustered transcription factor genes of an ethylene response factor (ERF) subfamily; in the nic2
133

mutant, at least seven ERF genes are deleted altogether.
Overexpression, suppression, and dominant repression experiments using transgenic tobacco roots showed both functional redundancy and divergence among the NIC2locus ERF genes. These transcription factors recognized a GCC-box element in the promoter of a nicotine pathway gene and specifically activated all known structural genes in the pathway. The NIC2-locus ERF genes are expressed in the root and upregulated by jasmonate with kinetics that are distinct among the members. Thus, gene duplication events generated a cluster of highly homologous transcription factor genes with transcriptional and functional diversity. The NIC2locus ERFs are close homologs of ORCA3, a jasmonateresponsive transcriptional activator of indole alkaloid biosynthesis in Catharanthus roseus, indicating that the
NIC2/ORCA3 ERF subfamily was recruited independently to regulate jasmonate-inducible secondary metabolism in distinct plant lineages. Possible applications with this class of transcription factors for desired metabolic engineering of medicinal plants will be discussed.
Gene-mediated alterations in
Nicotiana
alkaloid
1 metabolism and effects on primary and secondary metabolism in response to stress
Dalton, HL 1
Hamill, J 1
, Edwards, A 1 , DeBoer, K
Monash University, Australia
1 , Neale, A 1 ,
Increasingly, alkaloids are recognised as important nitrogen-containing defensive compounds, the synthesis of which may increase in response to wounding in order to provide protection from predators in native habitats. A good example is the production of toxic pyridine alkaloids, which are a feature of all species in the genus
Nicotiana and some other genera in the Solanaceae family. Wound- or jasmonate-induced alkaloid synthesis in Nicotiana species involves an increasingly well understood cascade of regulatory genes/proteins leading to elevated transcript and enhanced activity of a number of key alkaloid biosynthetic enzymes such as ornithine decarboxylase (ODC), arginine decarboxylase (ADC), putrescine methyltransferase (PMT) and quinolinate phosphoribosyltransferase (QPT) and an isoflavone reductase-like protein (A622) which is associated with the final stage of pyridine alkaloid biosynthesis.
Production of alkaloids in plants places demands upon vital resources, particularly nitrogen, and thus requires close co-ordination between primary and secondary metabolism. Using various Nicotiana species as experimental models, the present study focuses on the regulation of polyamine and pyridine nucleotide (1°) metabolism and alkaloid (2°) metabolism and the regulatory control mechanisms that operate under normal and stressed conditions. Antisense and RNAi-mediated down-regulation of genes involved in Nicotiana alkaloid biosynthesis allows not only quantitative and qualitative effects upon alkaloid metabolism to be determined, but also effects on activity of related genes and enzymes in primary and secondary metabolism. The capacity of such transgenic Nicotiana tissues to redirect nitrogen into primary metabolism and growth is of particular interest – particularly after exposure to abiotic and environmental stresses such as apex removal, leaf wounding, exposure
134 of plants to drought, elevated temperatures and high light. Recent results show that RNAi-mediated down regulation of ODC transcript levels in N. tabacum , causes a substantial change in the overall alkaloid spectrum of transgenic tissues. These alterations are particularly evident after wound-associated stress with diminished levels of nicotine and an increase in concentrations of anatabine – normally a minor alkaloid.
In separate experiments, RNAi-mediated down regulation of A622 led to plants with scarcely any alkaloids in leaf and young shoot tissues – even when growth is stimulated by apex removal which normally results in a large increase in alkaloid accumulation in newly formed young tissues. These plants were morphologically normal but showed slower growth rates and reduced capacity to recover from damage to aerial tissues. They also showed enhanced photosensitivity which may be associated with alterations in pyridine cycle metabolism.
1
The diverse roles of cyanogenic glucosides in plants
Jørgensen, K 1,3
Sánchez-Perez, R 2
, Kannangara, R 1
, Møllel, BL 1
, Bjarnholt, N 1 ,
University of Copenhagen, Plant Biochemistry
Laboratory, Denmark; 2 Departamento de Mejora
Vegetal, Centro de Edafoloia y Biologia Aplicada del
Segura-Consejo Superior de Investigaciones Cientificas,
Spain; 3 3VKR Research Centre 'Pro-Active Plants',
University of Copenhagen, Denmark
Cyanogenic glucosides are ancient bioactive plant constituents which upon cellular disruption of the plant tissue in which they are present release toxic hydrogen cyanide to deter herbivorous insects and pests.
Cyanogenic glucosides are produced from one of the amino acids valine, isoleucine, leucine, phenylalanine, tyrosine and cyclopentenyl glycine with alsoximes and cyanohydrins as key intermediates. The pathway is catalyzed by two membrane bound cytochromes P450, belonging to the CYP79 and CYP71 family and a soluble
UDPG-glucosyltransferase. During evolution cyanogenic glucosides have been recruited for additional function such as scavengers of reactive oxygen species, transporters of reduced nitrogen and as buffer compounds to balance primary metabolism. Cassava
( Manihot esculenta ) is a vegetatively propagated tropical crop with starch rich tubers which serve as a key stable food in Africa. Major deficits of cassava are low protein content in the tubers, rapid post-harvest tuber deterioration and high content of cyanogenic glucosides.
Careful processing of cassava roots is required to avoid the risk of acute or chronic cyanide intoxication.
Unfortunately, processing to remove the toxins typically results in loss of protein, minerals, and vitamins. Cassava produces the cyanogenic glucosides linamarin and lotaustralin from the amino acids valine and isoleucin, respectively. Young leaves are the main site of synthesis.
These cyanogenic glucosides are then transported and stored in the root tubers. Besides serving a protective role, the cyanogenic glucosides may also function as a source of reduced nitrogen. In transgenic cassava where expression of the genes encoding the first step in the synthesis are blocked using RNAi technology, the lines with the lowest content of cyanogenic glucosides exhibit a specific phenotype in vitro showing elongated

seedlings and poor leaf development . When grown in media with extra nitrogen the phenotypic difference is minor and the nearly acyanogenic plants show wild type phenotype when grown in greenhouse. These observations indicate a role for cyanogenic glucosides during the early developmental phase of cassava. In cassava and sorghum ( Sorghum bicolor ), the seedling synthesises the cyanogenic compounds during the first days of germination. In Sorghum the cyanogic glucoside content per plant peaks 3-4 days after germination and the diminishes due to endogenous turn-over without release of hydrogen cyanide. In contrast to cassava, bitter almonds ( Prunus dulcis ) store the cyanogenic glucoside amygdalin in the seed. When the seed germinate the content of cyanogenic glucosides are turned over and is used by the developing plant. Cyanogenic glucosides have in addition to being defence compunds important roles during plant development and in the interaction between plants and herbivorous insects and pathogens.
Introducing 'Resurrection Plants'
Don F. Gaff 1
1 School of Biological Sciences, Monash University,
Clayton, Australia
Most vascular plants succumb to drought when their water potential falls below –15MPa. Resurrection plants have desiccation-tolerant foliage that survives –300MPa or, in many species, even below –600MPa. Rehydration of their airdry leaves brings rapid recovery – usually within 24h. They can survive the airdry state for 6 months to 2 years, depending on the species and on the ambient relative humidity. Resurrection plants occur over a wide range of climates, latitudes, altitudes, insolations, topography, soils and rock substrates. The hydrated foliage of most resurrection plants is not desiccationtolerant: desiccation tolerance is induced by moderate water deficits – a process accompanied by changes in protein composition and an accumulation of protective substances. Desiccation tolerance exists in only a small proportion of monocot species and in a much lower proportion of dicot species in several unrelated families.
Desiccation-tolerant seed and/or pollen are found in almost all non-resurrection angiosperm species. It seems most likely that desiccation-tolerant foliage has evolved separately in several angiosperm families when extreme selection for improved protoplasmic drought tolerance acted on plants in shallow soils by producing expression of their seed/pollen desiccation tolerance mechanism in buds, then in young leaves and finally in mature leaves
(but never in senescent leaf tissue).
New insights into the role of nucleotides and polyamines during drought stress in
Arabidopsis mutants
Estavillo, G
Crisp, P 1
1 , Chan, K 1 , Phua, SY 1
, Pogson, B 1
, Pornsiriwong, W 1 ,
1 The Australian National University, Canberra, Australia
Plants respond to drought by adjusting gene expression and metabolite composition in order to cope with stress.
We have recently reported that a sulfur-related metabolite, PAP (3'-phosphoadenosine 5'-phosphate), increases during drought stress in Arabidopsis . PAP is an inhibitor of RNA processing enzymes and can alter gene nuclear gene expression. The Arabidopsis mutant alx8 presents altered drought response, and increased PAP levels, which regulate a novel ABA-dependent pathway to control stomatal closure. Using a combination of genetic and biochemical approaches, we investigated the role of PAP during drought, its interaction with ABA and how the subcellular pools of this metabolite could contribute stress responses. Preliminary results suggest that PAP levels are regulated by both ABA-dependent and independent mechanisms. Also, the subcellular location of this metabolite is also important in regulating the drought response. Additionally, using double and revertant mutants with altered levels of both PAP, putrescine and leaf morphology, we investigated the causes of the drought tolerant phenotype of the alx8 mutant. Our preliminary results indicate that polyamines, originally thought to be required for the drought response, are not essential in alx8, and that its leaf morphology is not the main contributing factor of its drought tolerance. This research sheds new light into the roles of metabolites, gene regulation and physiology of drought in plants.
Functional analysis of dehydration responsive genes
1 in the resurrection grass
Sporobolus stapfianus
Griffiths, C 1 , Blomstedt, C 1 , Hamill, J
Monash University, Australia
1 , Neale, A 1
Plant drought tolerance is an increasingly important issue in the current global climate and will remain an issue as global temperatures rise. The aim of this project is to identify the functions of two novel genes that are being transcribed during the dehydration of the resurrection grass S. stapfianus which exhibits extreme drought tolerance. The transcripts from these two genes, SDG3i and SDG4i, accumulate during dehydration particularly at relative water contents (RWC) of 59-40% and 39-20% respectively. This project looks at the phenotypic changes in transgenic Arabidopsis over-expressing these genes, intracellular localisation of the genes, and hormone regulation of the genes in S. stapfianus. Motifs present in the putative protein encoded by SDG3i suggest it may be a membrane channel protein associated with the peroxisome while the motifs in the protein encoded by SDG4i suggest it may have a function associated with chromatin remodelling. The creation of homozygous
Arabidopsis and rice over-expressing these genes is being used to provide clues on how SDG3i and SDG4i function. GFP fusion constructs indicate that SDG4i is localised to the nucleus. The over-expression of SDG4i in Arabidopsis has significantly increased osmotic stress tolerance in comparison to the wild type suggesting that
SDG4i may have a role in regulating desiccation tolerance in S. stapfianus. Furthermore the SDG4i transgenic Arabidopsis also appear to germinate and grow in the cold suggesting at SDG4i is aiding in the regulation of other stress responses. By looking at these genes it is hoped that the information gained on the desiccation tolerance of S. stapfianus can be applied to
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agriculturally important crop species such as rice, corn and sorghum and ultimately render a desiccationsensitive plant desiccation-tolerant.
The evolutionary and biosynthetic origin of cyclic peptides in sunflower seeds
Mylne, J 1
1 Institute for Molecular Bioscience, University of
Queensland, Australia
Sunflowers have universal appeal. Their seeds are popular in many countries both as confectionary and a source of high quality oil. Perhaps less well known is that they contain head-to-tail cyclised peptides that are braced by an internal disulfide bond. One of these, called SFTI, is 14-residues in size and an extremely potent inhibitor of trypsin (1). SFTI has promise as a drug lead in its own right with drug-like inhibition of the oncogenic protease matriptase (2) and has been proposed as an ultra-stable protein engineering scaffold (3). Its structure, ideal for protease inhibition, is used as a starting point for the design of serine protease inhibitors and recently a modified version was shown to specifically inhibit the protease KLK4, which is involved in prostate cancer (4).
In this talk I will describe the unusual biosynthetic route for SFTI, and an even smaller cyclic peptide SFT-L1.
They are not encoded by dedicated precursors, rather they arise as ‘extra’, unrelated products from much larger proteins as these get proteolytically matured. We have been using the model plant Arabidopsis to study this biosynthesis using modified precursor protein transgenes combined with endogenous gene knockouts. Analysis of the sequences for the SFTI and SFT-L1 precursor proteins suggests a conserved processing mechanism requiring the enzyme asparaginyl-endopeptidase, has been converged upon at least thrice by plants to generate cyclic peptides. Upon close inspection, the precursor
DNA sequences also provide some tantalising clues as to the genetic event that gave rise to these sunflower cyclic peptides. (1) Luckett, S. et al. 1999. (2) Long, Y.Q. et al.
2001. (3) Daly, N.L. et al. 2006. (4) Swedberg, J.E. et al.
2009.
Directed evolution and rational design of immune receptor proteins in the flax–flax rust pathosystem
Ravensdale, M 1 , Thrall, PH 1 , Ellis, JG 1 , Dodds, PN
1 CSIRO, Plant Industry, Canberra, Australia
1
Disease resistance in the flax–flax rust pathosystem is determined by a gene-for-gene recognition where flax immune receptors interact with rust effector proteins. In one such example, members of the AvrL567 effector protein family in flax rust interact with members of the L receptor protein family (L5, L6, and L7) in flax. The physical nature of these direct protein interactions has resulted in a co-evolutionary 'arms race' between these organisms. Mutational analysis of amino acid polymorphisms occurring between AvrL567 variants has
136 revealed a number of solvent exposed surfaces that contribute to recognition specificity, suggesting the presence of multiple interacting surfaces on the corresponding L receptor proteins. Directed evolution and rational design strategies have been applied in attempts to understand, and possibly improve, the recognition specificities of L receptor proteins. Chimeric and randomly mutagenized L proteins have been constructed via overlap PCR and error-prone PCR, respectively. Evaluation of these modified L proteins in the yeast-2-hybrid assay has identified specific regions within the leucine-rich repeat domain that mediate interactions with specific polymorphic residues found in
AvrL567 variants, and has also revealed evidence of extensive co-variation between domains and residues within L proteins.
Structural basis of R-protein mediated disease resistance in flax against flax rust
1
Williams, S 1 , Ve, T 1 , Bernoux, M 2
Dodds, P 2 , Kobe, B 1
, Anderson, P 3 ,
School of Chemistry and Molecular Biosciences,
University of Queensland, Brisbane, Australia; 2
Plant Industry, Canberra, Australia; 3
CSIRO
School of
Biological Sciences, Flinders University, Adelaide,
Australia
A plant’s ability to detect an invading pathogen and circumvent a subsequent disease state is essential for its survival. The pioneering work of Harold Flor, using the interaction between flax and flax rust, demonstrated that this ability to detect and resist the infection of a specific pathogen rests with two critical genes; a resistance (R) gene in the plant and a corresponding avirulence
(effector) gene in the pathogen (1). This, so called ‘genefor-gene’ model, has subsequently been shown to apply in many other plant-pathogen interactions and has spawned considerable research efforts directed towards understanding R protein-effector interactions and the consequential disease resistance response. Using the flax/flax rust pathosystem our research aims to further understand the molecular basis of R protein-effector interactions utilising structural and biochemical approaches. Flax R proteins encoding tri-domain proteins with a central nucleotide-binding domain (NB), a Cterminal leucine rich repeat (LRR) and a Toll-interleukin
1 receptor-like (TIR) domain at the N-terminus. We have previously shown that the flax R proteins, M and L6, interact directly with the flax-rust effectors AvrL567 and
AvrM, respectively (2,3) and have defined the structure of the AvrL567 effector protein (4). Here we report the crystal structure of the TIR domain from the flax R protein, L6, at 2.3 Å resolution. The structure reveals important differences from the structures of mammalian
TIR domains, and highlights three separate functionally important protein surfaces, involved in dimerisation, interaction with a downstream signalling partner, and regulatory intramolecular interactions, respectively. We have also determined the structure of the flax rust effector protein, AvrM, at 2.7 Å resolution. AvrM, which has no significant sequence similarity with proteins of known structure, has a novel L-shaped helical fold, with two chains forming a dimmer that create an unusual nonglobular shape. Furthermore, using a combined phenotypic and biochemical analysis of mutants made

within the NB domain of the flax M protein, we further define the molecular requirements for R protein activation. Collectively, these results bring us closer to understanding the molecular basis of R protein mediated plant disease resistance, a perquisite to the future engineering of novel resistance specificities in important crop species. (1) Flor 1971. (2) Dodds et al. 2006. (3)
Catanzariti et al. 2010. (4) Wang et al 2007.
Structural basis of function of plant proteins
Bond, C 1 , Fujii, S 2 , Stanley, W 3 , Small, I 1
1 School of Biomedical, Biomolecular and Chemical
Sciences, The University of Western Australia, Crawley,
Australia; 2 Centre of Excellence in Computational
Systems Biology, The University of Western Australia,
Australia; 3 ARC Centre of Excellence in Plant Energy
Biology, The University of Western Australia, Australia
Organelle gene expression involves a series of more or less coupled processes from transcription through transcript processing to translation. Many of the events that occur during these processes require sequence recognition by RNA processing factors. In plants, pentatricopeptide repeat (PPR) proteins are the most numerous of these factors, with a bewildering array of
450+ family members in all flowering plants studied to date (1,2,3). PPR proteins are found in lesser numbers in all eukaryotes with a mitochondrial genome, and probably play a similar role in organelle gene expression in plants, animals, fungi and protists (4). The sequencespecific RNA binding ability of these proteins is remarkable, given the wide range of different targets bound by different family members, but the molecular basis for target recognition is almost completely unknown (5). Research is hampered by the lack of any experimental data on the detailed structure of these proteins due to the difficulty of producing the proteins in large quantities in a soluble form. However, there is a huge amount of sequence information available from many fully sequenced plant genomes. We show that this sequence data is sufficient to construct a plausible structural model of the PPR motif, and of a tandem array of PPR motifs, using contact site prediction from covarying amino acid pairs. The methods we employed should work similarly well on similar protein families containing tandem repeats for which many sequences are available. Furthermore, the patterns of selection on one particular class of PPR proteins give strong indications about which amino acid residues within the PPR motifs determine RNA binding specificity (6). Based on this, we propose a model for how PPR proteins bind RNA. (1)
Small, ID, & N Peeters 2000. (2) Lurin, C, et al. 2004.
(3) O'Toole, N, et al. 2008. (4) Schmitz-Linneweber, C,
& I Small 2008. (5) Delannoy, E et al. 2007. (6) Fujii, S et al. (in press).
Overgrowth mutants of barley and wheat: new insights into the functions of DELLA proteins
Chandler, P 1
1 CSIRO Plant Industry, Canberra, Australia
Defined dwarf mutants of barley and wheat were treated with a mutagen, sown in the field, and the M2 generation screened for mutants with enhanced growth relative to their dwarf siblings, but which still retained the original dwarfing mutation. Many such ‘overgrowth’ mutants were isolated and characterised, the vast majority being due to new mutations in the principal DELLA genes of these species (Sln1 and Rht-1 for barley and wheat respectively). More than 30 new DELLA alleles have been identified, and this number will increase as more mutants are sequenced. Overgrowth mutants grow faster than their dwarf parent because they have enhanced GA signalling. We investigated whether GA responses other than growth are also affected in such lines. á-Amylase production by endosperm half-grains of barley is normally seen only in the presence of an active GA.
Overgrowth lines showed clear examples where the original parental response was maintained, but also other examples of moderate and of high á-amylase production in the absence of active GAs. Some lines with only a partial restoration of growth showed high levels of áamylase production. Analogous results were found in wheat, with a poor correlation between mature plant height and the maximum extent of coleoptile elongation.
Both sets of results indicate a considerable degree of specificity in the extent to which different overgrowth alleles influence different GA responses. In barley we have observed novel phenotypes associated with some overgrowth alleles. Several new alleles are associated with an increase in grain size of approximately 25%, averaged over several greenhouse generations. This effect has also been observed in back-crossed material in a commercial barley cultivar. A combination of two different overgrowth mutations results in barley lines that grow much better in the presence of exogenous abscisic acid (ABA) than either single mutants or the parental line. Such lines also had faster growth than either single mutants or the wild type when subjected to water deficit.
DELLA proteins contain several conserved amino acid sequence motifs, and the functions of these regions, which include interactions with other proteins such as the
GA receptor, an F box subunit involved in targeting
DELLA for degradation, and PIF (phytochrome interacting factor) transcription factors, are being actively studied in rice and Arabidopsis . The phenotypes of different barley and wheat overgrowth alleles will allow specific changes in DELLA motifs to be related to different roles of this protein in influencing phenotype.
These are likely to involve differences in binding to interacting proteins. In one case, different mutations of the same amino acid residue of the SLN1 protein in barley lead to clearly different phenotypes. DELLA semidwarfs formed the basis of the ‘Green Revolution’ in wheat, and are still used in the vast majority of wheat varieties. By generating many new semi-dwarfing alleles, potentially associated with other useful traits (early vigour, high grain dormancy, rapid á-amylase production), we hope to extend the range of semidwarfing alleles available for wheat and barley breeding.
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Diversification and mycorrhizal specificity of
1 flowering plants living on arbuscular mycorrhizal fungi
Merckx, V 1 , Smets, EF 1,2
Netherlands Centre of Biodiversity Naturalis (section
NHN), Leiden University, The Netherlands; 2 Laboratory of Plant Systematics, K.U.Leuven, Belgium
Fully mycoheterotrophic plants are achlorophyllous during their entire development and obtain all of their carbon from root-associated fungi. Most fully mycoheterotrophic flowering plants are growing in the leaf litter of dense tropical rain forest and associate with arbuscular mycorrhizal fungi (Glomeromycota). Due to the rarity and ephemeral occurrence of tropical mycoheterotrophic plants we know little about the evolution and ecology of the intimate interaction between these plants and their fungi. We use nuclear and mitochondrial DNA data to construct diversification hypotheses for mycoheterotrophic Burmanniaceae,
Thismiaceae, Triuridaceae, and Gentianaceae. Molecular clock inferences allow us to compare the timing of the shifts towards mycoheterotrophy in these different clades. Our results indicate that in some clades a fully mycoheterotrophic mode of life evolved relatively recently, while other clades share an ancient origin. Older mycoheterotrohic clades consist of considerable more species than more recent clades. This demonstrates that mycoheterotrophic lineages are able to persist and diversify over considerable periods of time. In addition, we sampled roots of a subset of mycoheterotrophic species from each clade and analyzed the small subunit ribosomal DNA gene amplified from root DNA extracts using primers specific for arbuscular mycorrhizal fungi.
The results indicate that certain clades of arbuscular mycorrhizal mycoheterotrophs consist of specialized species, that associate with very narrow lineages of fungi.
Other clades contain generalist species, that are able to associate with a broad range of fungi. Generalist mycoheterotrophic species seem to have broader distribution ranges than specialists.
Factors driving the mycoheterotrophic versus autotrophic carbon gain by green orchids
Gebauer, G
H 1
1 , Stöckel, M 1 , Preiss, K 1 , Adam, I 1 , Liebel,
1 BayCEER, Laboratory of Isotope Biogeochemistry,
University of Bayreuth, Bayreuth, Germany
In the mycorrhizal symbiosis plants usually exchange photosynthates for mineral nutrients acquired by fungi from the soil. This mutualistic arrangement has been subverted by a few hundreds of fully mycoheterotrophic plant species that lack the ability to photosynthesize. The most numerous examples of this behaviour are found in the orchid family. Non-photosynthetic orchids are known to be highly specialized exploiters either of the ectomycorrhizal symbiosis of trees or of wood-rotting saprotrophic fungi. These fully mycoheterotrophic
138 orchids are characteristically distinguished in their C and
N stable isotope natural abundance from co-occurring autotrophic plants in a similar way as various functional types of fungi are characteristically distinguished in their isotope signature from co-occurring plants (Gebauer &
Meyer 2003). Also among the numerous green orchid species more and more representatives, specifically from the subfamily Epidendroideae, turn out to possess isotope signatures positioned between fully autotrophic and fully mycoheterotrophic plants. These orchids live obviously partially at the cost of their fungal partners. Therefore, their type of nutrition is called partial mycoheterotrophy
(Gebauer & Meyer 2003). Here we present evidence that the proportional carbon gains from photosynthesis or from the fungal source by these partially mycoheterotrophic orchids are not constant. They are rather related to a couple of biotic and environmental factors. Of specific importance are (1) the functional types of fungal partners, (2) the light climate at the orchid habitat and (3) the leaf chlorophyll concentration.
Orchids associated with fungi simultaneously forming ectomycorrhizae with neighbouring trees gain considerably more carbon from the fungal source than closely related orchids associated with saprotrophic fungi of the polyphyletic Rhizoctonia group (Liebel et al.
2010). Even within orchid species the proportional carbon gain from either source can vary considerably.
Cephalanthera damasonium individuals living in the deepest shade of beech forests and individuals of the same species with variegated leaves and therefore having considerably less leaf chlorophyll concentrations tap considerably more on the fungal carbon source than individuals living in more sunny forest gaps and having fully green leaves (Preiss et al. 2010; Stöckel et al. 2010).
Our results document furthermore that the orchids’ change in proportional reliance on fungi as a carbon source affects leaf carbon and nitrogen concentrations.
Parallel evolution of mycoheterotrophy in land plants: a different story in the tropics vs temperate
1 regions?
Selosse, M-A 1 , Martos, F 1
Centre d'Ecologie Fonctionnelle et Evolutive, CNRS
Montpellier, France;
Basel, Switzerland
2
, Walder, F 2 , Courty, P-E 2
Botanical Institute, University of
The evolution of the land flora has provided repeated emergences of the mycoheterotrophic habit, where achlorophyllous plants exploit carbon from fungi mycorrhizal in their roots. This condition, suggested to be an adaptation to forest environments where little light is available, was mainly studied through two tools: fungal molecular barcoding allowed identification of the
(often uncultivable) fungi from the roots; natural isotopic abundances in mycoheterotrophs were instrumental in supporting that a given fungal guild was providing carbon to the plant. Most classical works investigated temperate and Mediterranean species that proved to have specific basidiomycetes fungal partners, forming the socalled ectomycorrhizae with surrounding trees. Recently, the research interest somewhat shifted to subtropical and tropical forests, where most mycoheterotrophic species are growing, including many species that connect to arbuscular-mycorrhizal (AM) fungi. Studies by our team and others have shown that high specificity for

mycorrhizal fungi is not the rule among tropical mycoheterotrophs. Although some species are specific, other species associated with basidiomycetes or AM fungi show less specificity, even if they remain selective
(i.e. have preferenda among the whole diversity of the targeted fungal guild). Secondly, especially in forest devoid of ectomycorrhizal basidiomycetes, other fungal guilds were targeted: some orchids from unrelated genera receive carbon from saprotrophic, wood-decaying or litter-decaying fungi, as supported by their isotopic abundances. At the same time and even in same forests,
AM fungi were used by other mycoheterotrophs, as stated above, e.g. in the Gentianaceae and
Burmanniaceae families. Our ongoing works suggest that, in the later case, the isotopic properties of the continuum between green plant (providing carbon) – AM fungi – mycoheterotrophic plant shows differences as compared to the analogous continuum for mycoheterotrophs associated with saprotrophic or ectomycorrhizal basidiomycetes. Moreover, C/N values, that are often low in mycoheterotrophs associated to basidiomycetes, are unexpectedly higher in AM associated mycoheterotrophs. Based on our recent data and several published ones, we hypothesize here that different factors contribute to the features recorded for tropical mycoheterotrophs. First, the ability of saprotrophs to support growth of mycoheterotrophs may simply be a question of water availability and duration of activity over the year. Indeed, in every biomes, saprotrophic fungi support a mycoheterotrophic growth during underground germination of orchid seeds, but this stops when plant enlarge and emerge from the soil, into a more desiccant environment. Second, AM-associated mycoheterotrophy may have evolved purely to support carbon need of the mycoheterotrophs, especially in the framework of shaded, but not N-limited tropical forests; at the opposite, the evolution of basidiomycetesassociated mycoheterotrophs may be linked to N acquisition in N-limited, but not always dark, temperate forests. Thus, we discuss that, beyond apparent similarities, the parallel evolution of mycoheterotrophy in land plants followed evolutionary pathways that are linked to the ecology of the respective plant lineages involved, thus differing from one biome to another.
Australian obligate myco-heterotrophic orchids and their fungal associations
Dearnaley, JDW 1 , Le Brocque, AF 2 , Bougoure, JJ
1 Australian Centre for Sustainable Catchments and
Faculty of Sciences, The University of Southern
Queensland, Australia;
Laboratory, USA
2 Lawrence Livermore National
All orchids rely on mycorrhizal fungi for provision of carbon compounds during seed germination and uptake of inorganic nutrients and water throughout the life cycle.
For adult green orchids, the flow of carbon appears to be reversed with photosynthate provided to fungal partners in exchange for their services. A small proportion of the approximately 30,000 orchid species worldwide are not photosynthetically active at adulthood. These obligate myco-heterotrophic orchids rely on mycorrhizal fungi to provide carbon compounds as well as inorganic nutrients and water throughout their life cycles. Our research has been focused on understanding the mycorrhizal biology of four Australian obligate myco-heterotrophic orchids, namely Dipodium variegatum, Dipodium hamiltonianum,
Erythrorchis cassythoides and Gastrodia sesamoides .
Identifying the mycorrhizal fungi of obligate mycoheterotrophic orchids is complicated as the fungi involved are usually impossible to isolate into pure culture. We have circumvented this problem by extracting DNA from whole orchid roots and separating fungal from plant DNA via PCR with fungal specific ITS primers. Amplicons have been cloned and sequenced and
GeneBank BLAST searches used to identify the fungal partners of the four orchid species. To support our interpretations of the results of these molecular analyses, we have used mass spectrometry to study natural carbon and nitrogen stable isotope abundances within orchid tissues. Fungal ITS sequencing suggested that the fungal community of Dipodium variegatum included Russula,
Trichoderma and Verticillium spp. The fungal community of Dipodium hamiltonianum consisted of
Gymnomyces , Russula and Penicillium spp. Analysis of the fungal endophytes of Erythrorchis cassythoides suggests that the orchid is colonized by both ectomycorrhizal fungi such as Russula and Coltricia as well as the saprotrophic Gymnopus . In Gastrodia sesamoides , the main fungal partner revealed by ITS sequencing was a member of the wood-rotting
Campanella genus. This finding was reinforced by stable isotope data which suggested that the orchid receives carbon from saprotrophic fungi. These results suggest that like North American and European obligate mycoheterotrophic orchids, some Australian obligate mycoheterotrophic orchids are colonised by members of the
Russulaceae. As these fungi are ectomycorrhizal and
Dipodium species typically grow at the base of
Eucalyptus , the orchids may be indirect parasites on the trees but this remains to be proven. The involvement of
Gymnomyces spp. as mycobionts in D. hamiltonianum may be one reason why this orchid is becoming rare as the fruiting bodies of this species are much sort after food of fungivorous marsupials which may act as dispersal agents for fungal spores. The occurrence of both ectomycorrhizal and saprotrophic fungi as mycobionts in
Erythrorchis cassythoides suggests that the orchid may be able to survive the death of its host tree by switching from a parasitic mode of nutrition to a saprophytic one.
Gastrodia sesamoides is exceptional among the four orchids studied in that it appears to exclusively receive its carbon from a wood-rotting saprotrophic fungus.
Investigation of the mycorrhizal biology of these
Australian obligate myco-heterotrophic orchids has thus revealed some novel orchid-fungal associations as well as provided insight into the multiple nutritional modes of these unusual plants.
Comparative plastid genome study in
1 mycoheterotrophic Ericaeae
Stefanovic, S 1 , Braukmann, T 1
University of Toronto,Mississauga, Canada
Heterotrophic plants show a wide range of evolutionary degradation of photosynthetic capability and rely entirely or partially on their host autotrophic plants to supply water and nutrients. These plants are usually divided into two morphologically distinct yet evolutionary artificial groups, parasitic plants and mycotrophic plants,
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depending on whether they derive all or part of their nutrients via direct connection with the host plant tissue
(haustorium), or via symbiotic relationship with mycorrhizal fungi, respectively. Haustorial parasitism has evolved at least 11 times independently within flowering plants and there are at least 10 independent origins of mycoheterotrophy in angiosperms plus one in conifers. Each of those lineages of heterotrophs represents an independent natural genetic experiment whose plastids and genes have evolved under relaxed functional constraints and therefore, each represents a unique opportunity to dissect plastome function and evolution. While several of haustorial parasitic groups have been moderately- to well-studied (e.g., Epifagus ,
Orobanchaceae; Cuscuta , Convolvulaceae), very few of mycoheterotrophic lineages were subjects of any significant plastid genome study to date. Hence, little is known about their plastid genome size, content, gene order, and expression. Ericaceae, the heather family, is a large and diverse group of plants known to have elaborate symbiotic relationships with fungi
(mycorrhizae). Grounded in an improved phylogenetic framework and a broad taxonomic sampling, a comparative investigation of plastid genomes was conducted in this family using a Southern hybridization approach. This survey encompassed lineages within
Ericaceae that have quite different life histories and trophic levels, including multiple representatives of hemi-mycoheterotrophs (pyroloids) and holomycoheterotrophs (monotropoids). Also, a number of fully photosynthetic taxa, carefully chosen to best represent the other major clades within the family, were included. More than 50 probes, derived from all categories of protein-coding genes usually found in plastomes of fully photosynthetic plants, were used.
Comparative analyses of the plastomes of plants along the full trophic spectrum, from autotrophy to hemiheterotrophy to full heterotrophy, allow us to assess the degree to which genomic changes take place in hemiheterotrophs prior to complete loss of photosynthesis and to dissect the evolutionary pressures on plastomes exerted by other plastid metabolic functions. Also, comparisons with well-studied parasitic systems provide us with an opportunity to assess whether haustorial parasitism and mycoheterotrophy have different genomic consequences.
Pylogenetics of mycoheterotrophic monocots and gymnosperms using plastid loci
Lam, V 1 , Biffin, E 2 , Rai, H 3 , Chang, Y 1 , Graham, SW 1
1 University of British Columbia, Canada;
Adelaide, Australia; 3
2 University of
Utah State University, USA
Mycoheterotrophic plants obtain their carbon budget from fungi involved in mycorrhizal networks, and thus indirectly from green plants. Full mycoheterotrophs are not photosynthetic and they often have reduced, scalelike leaves and unusual floral biology. Mycoheterotrophy has arisen multiple times in plant evolution, most abundantly in the monocots (i.e., in Burmanniaceae,
Corsiaceae, Iridaceae, Orchidaceae, Petrosaviaceae,
Thismiaceae and Triuridaceae); this nutritional mode is known only sporadically in other land plants, including the only heterotrophic conifer, Parasitaxus usta .
Genomic consequences of the transition from an
140 autotrophic to fully heterotrophic lifestyle are evident in the loss or degradation of photosynthesis-related plastid genes, such as the Rubisco large subunit ( rbcL ).
Therefore, phylogenetic studies of these plants have tended to focus on mitochondrial or nuclear markers, either to study relationships within taxa, or to place them in overall plant phylogeny. Some of these placements, based in part on retained plastid genes, are now uncontroversial. For example, Petrosavia is now recognized as one of two genera of Petrosaviales, one of the deepest diverging lineages in monocot phylogeny.
However, the local phylogenetic placement of other mycoheterotrophic taxa is less clearly resolved, and in general these plants remain poorly integrated with mainstream plant phylogenetic studies, which continue to focus on plastid markers. Essential, non-photosynthetic genes in the plastid genome may be retained regardless of trophic lifestyle, and represent potentially useful sources of phylogenetic markers. They may also provide insights into plastid genome evolution and function in heterotrophic plants. Here we report on our attempts to recover plastid genes involved in different aspects of plastid function: accD (acetyl-CoA carboxylase, involved in fatty-acid biosynthesis), mat K (the plastid Group II intron maturase) and clpP (serine protease involved in hydrolysis of peptide bonds). We surveyed these genes in a broad range of monocots, including the major mycoheterotrophic lineages outside Orchidaceae. We recovered at least one of these genes from the mycoheterotrophic families, including the first plastid sequences reported from Corsiaceae (for Arachnitis uniflora ) and Triuridaceae ( Sciaphila spp.). Phylogenetic analyses were complicated in some cases by long branches subtending sequences from mycoheterotrophs, but in most cases the gene products were open reading frames. Our data are consistent with placements of
Burmanniaceae and Thismiaceae in Dioscoreales, and indicate a (tentative) placement of Corsiaceae in Liliales.
Sequences recovered from Sciaphila are both diverse and divergent, but some of them place this taxon within
Stemonaceae (Pandanales). We also surveyed a broad range of plastid genes in the mycoheterotrophic (and hemiparasitic) conifer Parasitaxus , to place it better in conifer phylogeny. We recovered open-reading frame sequences from plastid genetic apparatus genes (accD, ribosomal protein genes, RNA polymerase subunit genes and mat K). Most photosynthesis-related genes
(photosystem II subunit genes) had interrupted reading frames and are present as multiple recent and divergent copies, but atpB and atpE are present as open reading frames, hinting at possible retention of function. We were able to obtain a preliminary estimate of its divergence of
Parasitaxus from two conifers ( Manoao and
Lagarostrobos ) that we were also able to confirm as its closest relatives in Podocarpaceae.

Evolutionary footprints of higher plants in chemical space
Backlund, A 1
1 Uppsala University, Sweden
Natural compounds and their biosynthesis machinery are developed and honed under evolutionary pressure, displaying a unique diversity of chemical properties and corresponding biological activities. The width of their chemical diversity when compared to various combinatorial chemistry compund libraries, have in the last few years fostered a renewed interest in natural compounds also from e.g. the pharmaceutical industry.
Of utmost importance for a rational characterization of the observed diversity of plant secondary compounds are two aspects: one the identification and charting of the biologically relevant chemical space, the other a similar charting of the corresponding evolutionary space (1, 2 pro parte). The first key to achieve such a goal would be to explore evolutionary space by elucidating and utilising robust phylogenies for the organisms under study. From this basis reflecting the evolutionary history and hence biosynthesis development, further conclusions can be drawn (e.g. 3). The second key would consequently be the coverage of secondary metabolites' chemical space.
For this purpose we introduced ChemGPS-NP (4, 5), with the aim to provide a tool for more efficient and stringent compound comparison, to identify parts of chemical space related to particular biological activities, and to track changes in chemical properties due to e.g. evolutionary traits and modifications in biosynthesis.
Physical-chemical properties not immediately discernible from structural data can be compared. Based on these initial assumptions, the intersection of chemical and evolutionary space have been explored. With regard to e.g. compound classes such as iridoids, betalains, and sesquiterpene lactones, evolutionary patterns of changes in physical-chemical properties are observed and compared. For eight major classes of plant defence peptides analyses of structure base alignments provide arguments for rational classification. References: (1)
Bohlin, L et al. 2007. (2) Bohlin, L et al. 2010. (3) el-
Seedi, H et al. 2005. (4) Larsson, J et al. 2005. (5)
Larsson, J et al. 2007.
The suitability of various classes of natural products as chemosystematic markers: case studies from the
Cichorieae and Gnaphalieae (both Asteraceae)
Zidorn, C 1
1 University of Innsbruck, Austria
Chemosystematics is an interdisciplinary field of natural sciences investigating the distribution of natural products within all groups of living organisms (archaea, bacteria, fungi, plants, and animals). Many primary metabolites are shared by most forms of life (with the notable exception of many archaea) and studies in chemosystematics aiming at the elucidation of relationships between taxa are therefore usually focused on secondary metabolites. Recently, the systematics of the Hypochaeridinae subtribe (Asteraceae, Cichorieae) was re-evaluated based on a combination of published and new molecular data as well as on a combination of these data with phytochemical data. The results required major changes in the systematics of the subtribe. The overall satisfactory level of knowledge of the phytochemistry of the various genera of the
Hypochaeridinae (and the Cichorieae in general) together with the good coverage of molecular data gave the opportunity to systematically compare the degree of systematic information extractable from the distribution patterns of plant secondary metabolites from different classes of natural products. Three classes of secondary metabolites were analyzed, caffeic acid derivatives, flavonoids, and sesquiterpene lactones. Flavonoids are due to the facts that they are easily detectable by simple means, occur ubiquitously in higher plants, and show a high degree of structural variations, one of the favorite classes of secondary metabolites for phytochemical investigations. However, the erratic distribution of some groups of flavonoids, e.g. isoetin derivatives, makes flavonoids suitable systematic markers only at low levels like species and intra-specific level. Sesquiterpene lactones on the other hand are especially in the
Asteraceae excellent chemosystematic markers at the level of tribes, subtribes, and genera. These patterns were also verified for the Hypochaeridinae. In recent studies, the formerly overlooked suitability of caffeic acid derivatives as excellent chemosystematic markers was established for some genera of the Cichorieae, e.g. the morphologically similar genera Crepis and Hieracium are characterized by the presence and absence, respectively, of the caffeoyl tartaric acid derivatives. The alpine Leontopodium alpinum (Edelweiss) features unique caffeoyl glucaric acid derivatives. A phytochemical investigation of European members of the closely related genus Gnaphalium revealed that caffeoyl glucaric acid derivatives currently known only from
Leontopodium are also present in Gnaphalium .
Interestingly, Gnaphalium species ecologically restricted to high alpine environments contain much higher concentrations of these radical scavenging compounds than their lowland relatives. This hints to a major caveat of all chemosystematic studies, the level of many natural products is directly linked to ecological stressors and the analytical absence of a certain secondary metabolite does not necessary imply the inability of the investigated plant taxon to synthesize these compounds, if need be. Taking other related studies into account, a picture emerges, where chemical characters are in their suitability as systematic markers as well as in their dependency from ecological factors similar to morphological characters.
The applicability of various kinds of these morphological markers, too, differs depending on the group investigated; one or the other set of characters might be better suited to characterize taxa at different systematic levels and in different systematic groups.
Shionone synthase, an oxidosqualene cyclase of an
Asteraceae plant that yields a unique tetracyclic triterpene ketone: molecular cloning and evolutionary view
Ayabe, S-I 1 , Sawai, S 1
T 1 , Akashi, T 1
, Uchiyama, H
, Takahashi, T 1
1 , Mizuno, S 1 , Aoki,
141

1 Dept of Applied Biological Sciences, Nihon University,
Japan
Plant triterpenoids contain both primary metabolites, i.e., the precursors of membrane sterols and steroid hormones, and secondary metabolites like pharmaceutically important saponins and ecophysiologically active chemicals. They display a remarkable variety with about 100 mono- to penta-cyclic skeletons, but structures most frequently found are
6/6/6/5 tetracycles and both 6/6/6/6/6 and 6/6/6/6/5 pentacycles. The skeletal variety is attributed to both the diversity of oxidosqualene cyclase (OSC) enzymes
(genes) and multiple products from single OSC reactions.
Molecular phylogenetic analysis indicates that plant
OSCs belong to four families: lanosterol synthase (LAS), cycloartenol synthase (CAS), lupeol synthase (LUP) and the most divergent
β
-amyrin synthase (BAS) families.
Shionone, the major triterpenoid of the roots of Aster tataricus , an Asteraceae plant used in traditional Oriental medicine, has a unique 6/6/6/6 tetracyclic skeleton with a
3-oxo-4-monomethyl partial structure. The A-C ring part of shionone is identical to the same part of a 6/6/6/6/6 pentacyclic triterpene, friedelin. The occurrence of shionone is very limited, whereas friedelin is widely distributed in land plants. While the biogenetic consideration was useful in the structural determination of shionone in the 1960s (1), no actual biosynthetic studies have been performed, and whether a single OSC carries out the construction of the rare skeleton and extensive methyl and hydride shifts to yield shionone or more than one enzymes are involved in its biosynthesis remains unclear. Only two OSCs have so far been reported to yield the 6/6/6/6 tetracycles; Arabidopsis thaliana baruol synthase (2) and Stevia rebaudiana baccharis oxide synthase (BOS; 3). In 2010, the first friedelin synthase cDNA was cloned from Kalanchoe daigremontiana (4), which should catalyze up to 10 rearrangement steps from 2,3-oxidosqualene. These
OSCs are all multiproduct enzymes, but shionone has never been documented as a product from their reactions.
In the present study, we have identified the cDNA of an
OSC that yields shionone for the first time. An OSC cDNA isolated from A. tataricus by PCR techniques was expressed in the LAS-deficient yeast, and the OSC reaction products accumulated in the yeast were identified. The enzyme yielded ca. 90% shionone and small amounts of byproducts, and was designated shionone synthase (SHS). The presence of 4-epishionone among the byproducts indicates that SHS catalyzes the production of the enol form of shionone, and subsequently the major product shionone and minor 4epishionone should be formed non-enzymatically.
Phylogenetic analysis and comparison of reaction specificity of SHS and other OSCs showed that SHS and
BOS have evolved in the Asteraceae from BAS lineages, and SHS must have acquired a high product-specificity and unique species-specificity in A. tataricus . The evolutionary and chemotaxonomic discussion incorporating recently reported plant OSCs yielding shionone-related triterpenes will be presented. (1)
Takahashi T et al. 1967. (2) Lodeiro S et al. 2007. (3)
Shibuya M et al. 2008. (4) Wang Z et al. 2010.
Biosynthesis and the systematic placement of
1
Gelsemiaceae
Backlund, M 1 , Larsson, S 2 , Backlund, A 3
Swedish Museum of Natural History, Stockholm,
Sweden; 2 Royal Botanical Gardens, Kew, UK; 3 Uppsala
University, Uppsala, Sweden
The order Gentianales has during the last decade been the target for a number of major systematic endeavours, most based on molecular data from DNA sequences. Although a number of achievements have been made, including the interfamilial systematics of Rubiaceae (1, 2), the reduction of Asclepiadaceae as a subfamily of
Apocynaceae (3), and the re-circumscription of
Loganiaceae (4). Among the remaining interfamilial issues remaining to be addressed in the Gentianales, we find the question of the systematic rank and affinities of the family Gelsemiaceae (5). The small family
Gelsemiaceae, comprised of only the two genera
Gelsemium and Mostuea , and their 17 species, exhibit morphological similarities with both Loganiaceae s.str. and Apocynaceae. Also from phylogenetic analyses of
DNA sequence data have no conclusive results yet been reached, with only weak affinities indicated. There is, however, additional information derived from analyses of alkaloid biosynthesis which may shed further light on this situation. From the Gelsemiaceae a set of unique alkaloids have been described. Comparing core structural elements from these, with known biosynthetic reactions and alkaloids from the neighbouring families
Apocynaceae and Loganiaceae provide a plausible explanation for the ancestry of the Gelsemiaceae alkaloids. This demonstrates yet another example of how interpretations of natural products biosynthesis may aid in systematic classifications or circumscriptions.
References: (1) Bremer, B 1996. (2) Bremer, B et al.
1995. (3) Sennblad, B & Bremer, B 1996. (4) Oxelman,
B et al. 1999. (5) Backlund, M et al. 2000.
Can phylogenies help bioscreening? A comparative analysis of the medicinal floras of Nepal, New
Zealand and the Cape of South Africa
Saslis-Lagoudakis, CH 1,2 , Savolainen, V 2 , Hawkins, JA 1
1 Dept of Biological Sciences, The University of Reading,
Reading, UK; 2 Dept of Life Sciences, Imperial College,
London, Silwood Park Campus, Ascot, UK
Several ethnomedicinal studies have aimed to reveal relationships between ethnomedicinal floras of different peoples. Traditionally, such efforts have been impeded due to the fact that different peoples are exposed to different local floras, limiting comparability between dissimilar floras. However, the advent of molecular phylogenetics in recent decades has revolutionised our knowledge of plant relationships and has provided a framework where for a number of plants a phylogenetic distance can be calculated. This allows us to calculate relatedness of ethnomedicinal species that are used in different parts of the globe, in order to quantify how
‘phylogenetically different’ ethnofloras from different cultures are. In this study, the phylogenetic relationships of the floras from three distant regions from biodiversity hotspots (Nepal, New Zealand, and the Cape of South
142

Africa) were reconstructed. Trees were constructed for individual floras as well as for the three floras combined.
Ethnomedicinal usage from each region was superimposed on the phylogenetic trees. We applied techniques initially designed for community ecology to calculate phylogenetic distance of the ethnofloras and to highlight lineages that are more heavily used than others.
We found significant phylogenetic clumping for medicinal plants in the phylogeny in most cases in the individual trees and in some cases in the combined tree, suggesting that i) certain lineages are richer in medicinal properties than others and ii) close relatives are used in different regions to treat similar conditions. Usage of closely related taxa globally would indicate that these taxa are the most promising in delivering new medicines, as they have been selected independently for similar uses. We are proposing that these techniques can be applied to guide bioscreening from phylogenetic data and the same phylogenetic manipulations can be used at the interface of phylogeny and phytochemistry. This study provides a methodological and conceptual framework of how phylogenies, combined with ethnomedicinal data, can help bioscreening programmes and reveals that ethnomedicinal floras around the globe might not be as distantly related as previously thought.
The predictive power of phylogenies for traditional medicine and lead discovery in Amaryllidaceae
1
Rønsted, N
Jäger, A 1
1 , Stafford, G 1 , Meerow, A 2
University of Copenhagen, Denmark;
, van Staden, J 4 ,
Tropical Garden, Miami, Florida, USA;
2
Dept of Agriculture, Miami, Florida, USA;
4
United States
3 Fairchild
University of
KwaZulu-Natal, Pietermaritzburg, South Africa
Plant secondary metabolites are produced and selected by evolution for their biological activity. Such natural products have always played a major role in traditional medicine and as leads for modern medicine. Only a small fraction of the world’s biodiversity has been explored for chemical and biological activity. A correlation between phylogeny and biosynthetic pathways is often assumed and could offer a predictive approach enabling more efficient selection of plants for traditional medicine lead discovery. However, formal tests of correlations between phylogeny and chemistry are rare, and the potential predictive power is consequently unknown. As a case in point, we are exploring the Amaryllidaceae subfamily
Amaryllidoideae sensu APG, which is known for subfamily specific alkaloids with activity in the central nervous system (CNS). Galanthamine registered for treatment of Alzheimers disease was first isolated from the Caucasian snowdrop Galanthus woronowii . We present a phylogenetic hypothesis of the Amaryllidaceae subfamily Amaryllidoideae based on nuclear and plastid
DNA sequences of 120 of the circa 850 species, representing all tribes and geographical regions. All major lineages are now well supported and the extended sampling uncovered several genera as non-monophyletic, emphasizing the importance of using phylogenetic rather than classical classification for interpretation of character distribution. Alkaloid profiles and CNS-related bioactivity profiles are significantly correlated with phylogeny using formal tests. Relationships between phylogenetic and chemical diversity are further explored.
The predictive power is used to select candidate taxa for lead discovery and to make recommendations for traditional use.
Plants as concept generators for biomimetic materials and structures with hierarchical organisation
Speck, T 1
1 Plant Biomechanics Group Freiburg, Botanic Garden,
Faculty of Biology, University of Freiburg, Germany
In the course of evolution, plants have developed a hierarchically organisation on at least five levels: in the stem, the tissues, the cells, the cell wall structure and the biochemical level. These hierarchies are also expressed by gradients in the tissues and the presence of sublevels
(e.g. groups of fibrous bundles-fibrous bundles-fibres) and hold responsible for beneficial mechanical properties like a benign fracture behavior, an optimized vibration damping or impact damping, high bending stiffness, heat insulation, avoidance of notch stresses and delamination as well as actuation and self-adaptation properties.
Therefore, analysing the hierarchical organisation in plants is a major key toward understanding the morphological-mechanical relationships in the natural structures and for assessing the potential for technical implementation. Over the last few years, it has been shown in a multitude of biomimetic projects that such a translation into technical products is feasible. However, the hierarchical organisation that is found in the biological concept generators has only been incorporated in a small part of technical materials and structures. As a result, (1) the identification of the biomimetic potential by functional morphology and biomechanics and (2) the abstraction of the identified principles remains one of the future challenges for developing innovative products.
Development of optimised biomimetic shells and branchings based on biological concept generators
Masselter, T
Speck, T 1
1 , Seidel, R 1 , Haushahn, T 1 , Thielen, M 1 ,
1 Plant Biomechanics Group Freiburg, Botanic Garden,
Faculty of Biology, University of Freiburg, Germany
Hierarchical organisation is present in most plants on at least five hierarchical levels: stem, tissue, cell, ultrastructural and molecular level. This morphological structuring has important implications on the mechanical properties of these plants, e.g. lightweight, optimised stiffness, damping and benign fracture behaviour.
Furthermore, it could also be shown that the morphological and mechanical optimisations present in biological organisms can be abstracted and be used to develop optimised bio-inspired technical products. Two recent biomimetic projects of the Plant Biomechanics
Group in Freiburg are dealing with the damping behaviour of nuts and drupes resp. the branchings of arborescent monocotyledons and arborescent cacti and
143

their potential for the development of biomimetic products. In the first project, the damping properties of the Macadamia nut, the Pumello and the coconut are investigated to gain a better understanding of the morphological-mechanical relationships in these fruits.
The tough seed coat of the Macadamia nut serves as an excellent biological role model for puncture resistant materials. The hierarchically organised, thick and spongy fruit wall of the pummelo, Citrus maxima , allows for high dissipation of the initial potential energy during impact. The coconut, Cocos nucifera , combines both hierarchical structures in a sandwich with an outer fibrous layer capable of high energy dissipation by large deformation together with a tough hard inner shell.
Implementing the hierarchical structuring of the protection layer and the use of gradients together with fiber reinforcement is intended to lead to the development of biomimetic impact resistant composite materials for helmets, containers with high shock absorbance as well as space station protection against meteoroid impact and protection of vehicles. In the other project, the potential of hierarchically structured plant ramifications as concept generators for innovative, biomimetic branched fibre-reinforced composites was assessed by morphological and biomechanical analyses.
Promising biological role models were found within monocotyledons, i.e. Dracaena and Freycinetia as well as within columnar cacti like Cereus and Myrtillocactus .
These plants possess ramifications with a pronounced fibre matrix structure and a special hierarchical stem organization, which markedly differs from that of other woody plants by consisting of isolated fibres and/or wood strands running in a partially lignified ground tissue matrix. The angles of the Y- and T-shaped ramifications in the plants resemble those of the branched technical structures. Our preliminary investigations confirm that the ramifications possess mechanical properties interesting for technical applications, such as a benign fracture behaviour, a good oscillation damping caused by high energy absorption, and a high potential for lightweight construction. Potential technical applications are hubs of wind-power plants, branch points of framework constructions in building industry, aerospace, ramified vein prostheses in medical technology or the connecting nodes of axel carriers. The results within the two projects demonstrate the high potential for a successful technical transfer and has led to the development of concepts for producing demonstrators in lab-bench scale that already incorporate ‘solutions inspired by nature’.
Biomechanical diversity of tropical rain forests as a basis for bio-inspired materials and structures
Rowe, N 1 , Paul-Victor, C 1 , Gueroult, M 1
1 Botany and Computational Plant Architecture, AMAP,
CNRS, University of Montpellier 2, France
Tropical rain forests are probably the most structurally complex ecosystems in the world and comprise a vast range of plant life forms and growth forms. Herbs, shrubs, trees, vines, lianas, epiphytes and hemi-epiphytes may all live in close physical contact and have adapted a wide range of biomechanical strategies for life in the forest. Two main factors potentially underline many of the mechanical strategies adopted by plants in the
144 tropical forest; first the need to reach light but at the same time survive in a mechanically perturbed environment with frequent tree-falls and debris-falls.
Second, the fact that different plant groups have different basic developmental constraints: such as woody dicots and non-woody monocots. Different growth forms have different mechanical constraints and have adapted many ways of producing stiffness, flexibility and resistance to failure that can offer a rich source for bio-inspired technologies. We survey some of the biomechanical strategies of climbing plants that are relevant to developing bio-inspired materials and structures. Woody lianas can develop what can be viewed as super gradient materials with extreme transitions in stiffness and resistance to failure. Monocotyledonous climbers such as the Ischnosiphon centricifolius (Marantaceae) develop high stiffness of cane-like internodes but also highly compliant branch points at the nodes that ensure high flexibility and resistance to failure in the event of mechanical perturbation. Attachment devices of nonwoody and woody climbers provide a wealth of mechanical strategies for bio-inspired applications including 'dumb-hooks' of climbing palms that need tension, friction or sharpness to remain attached to the support and 'smart hooks' of woody climbing species of
Bauhinia (Caesalpiniaceae) and Strychnos (Loganiaceae) that can eventually develop strong permanent tendrillike attachments but can first develop flexible 'clip on – clip off' hooks in mechanically perturbed environments.
Self-irrigation by fog collection: lessons from plant structures
1
Roth-Nebelsick, A 1 , Ebner, M
V 4 , Konrad, W 2
2 , Voigt, D 3 , Gottschalk,
2
State Museum of Natural History Stuttgart, Germany;
University of Tübingen, Institute for Geology, Tübingen,
Germany; 3 Functional Morphology and Biomechanics,
Zoological Institute, Christian-Albrechts University,
Kiel, Germany; 4 Institute for Textile Technology and
Process Engineering Denkendorf, Dept of Technical
Textiles, Denkendorf, Germany
Fog and dew collection as a substantial water source has gained increasing attention and there are ongoing efforts to optimize technical harvesting systems. Since many plants (and animals) benefit from fog and dew collection, the analysis of these biological systems can provide important information about useful strategies for obtaining water in arid environments. One important aspect in this respect is the role of surface properties.
Usually, a surface is characterized with respect to its interaction with liquids by the contact angle and/or water repellency which itself depends on the minimum drop size. Fog and dew, however, involve very small droplets in the range of a few tens of micrometers diameter. Since parameters such as contact angle and shape of the drop contact line depend in the case of rough surfaces on the ratio between drop size and size of the surface irregularities, surface structures can affect fog and dew collection via microdroplet behavior, the sticking/detachment of these drops and by directed water flow. A particular example is provided by the fogcollecting grass species Stipagrostis sabulicola that occurs exclusively on sand dunes of the Central Namib
Desert. S. sabulicola is able to harvest 4–5 liters per

nocturnal fog event and subsequently conducts the water almost entirely towards the plant base. In this manner, local soil moisture is concentrated and becomes available for the roots. The directed water conduction is due to a combination of various leaf surface traits. The natural system of S. sabulicola and its transfer into a technical fibre is described.
Biomechanics and functional morphology of suction traps in aquatic carnivorous bladderworts
(
Utricularia
spp.) as concept generator for biomimetic
1 products
Poppinga, S 1,2 , Weisskopf, C
Quilliet, Catherine 3
1 , Vincent, Olivier 3
, Joyeux, M 3 , Marmottant, P 3
Masselter, T 3 , Speck, T 1,2
,
,
Plant Biomechanics Group Freiburg, Botanic Garden,
2
Faculty of Biology, University of Freiburg, Germany;
Competence Network Biomimetics, Baden-Württemberg,
Germany, and BIOKON International-The Biomimetics
Association; 3 Laboratoire de Spectrométrie Physique,
University of Grenoble, France
For nutrient supply, aquatic carnivorous bladderworts
( Utricularia spp., Lentibulariaceae) catch small prey animals with suction traps. These traps, also called bladders, are considered as one of the most complex plant structures and show one of the fastest movements generally known from plants, which is particularly intriguing as it functions underwater. In collaboration between physicists from Grenoble and biologists from
Freiburg three species have been analysed as to their functional morphology, the biomechanics of trap and trap door, and the hydrodynamics involved. The bladders contain water which is pumped out by special glands, hence generating negative pressure und allowing elastic energy to be stored in the trap walls. A door with four protruding trigger hairs provides a watertight closure of the trap. When these hairs are touched by prey, the door opens and closes in a fraction of a second, and relaxation of the trap walls leads to the sucking in of water and prey. Prey animals are sucked in with an acceleration of up to 600 times that of gravity, leaving them no chance to escape. The door deformation involves a complete inversion of curvature which runs in several distinguishable intermediate steps. This ultra-fast, complex and at the same time precise and highly repetitive movement is enabled by certain functionalmorphological adaptations. Based on quantitative analyses of functional morphology and biomechanics of bladderwort traps, a transfer into innovative biomimetic products including micro-pipettes and microfluidic devices is intended.
The plant cuticle: multifunctional interface and model for innovations in engineering
Neinhuis, C 1 , Bargel, H 2 , Koch, K 3
1 Technische Universität Dresden, Germany;
Bayreuth, Germany; 3
2 Universität
Hochschule Rhein-Waal, Germany
A thin extracellular membrane, called the cuticle, covers the aerial surfaces of plants. It serves as the outermost border of plants and represents a composite consisting of three major components: the biopolyester cutin, soluble lipids of various chain lengths (waxes), and cellulose fibrils. The latter tightly connect the cuticle to the cell wall resulting in a gradient of material properties. The cuticle is a multifunctional interface that primarily prevents the plant from uncontrolled water loss, but also serves as a protective layer against biotic and abiotic environmental influences. In virtually all species of land plants, waxes cover the cuticle either as defined crystalline structures, thin films or thick layers. These layers or three-dimensional structures may grow up to several micrometers. The transport of lipids through the cuticle as well as the formation of epicuticular wax crystals based on self-assembly upon the surface is studied by atomic force microscopy, which allows observing the regeneration of defects of the outer surface wax coverage in situ. After removing the existing epicuticular waxes from leaves of different species, most plants instantaneously regenerate a multi-layered wax film as well as wax crystals. Considerable differences were found in the intensity of regeneration, as well as in the growth mechanisms. Based on AFM examinations we demonstrate that wax regeneration is a highly dynamic and rather fast process, indicating an overall importance of a continuous outer hydrophobic coverage.
These studies of the dynamic processes on the molecular level on the living plant surfaces may generate valuable insight in self-assembly processes that eventually can be applied in creating self-assembled structures in situ. Selfassembly processes should be preferred since structures are formed in a highly parallel manner independent of the surface geometry. Applications have been demonstrated earlier since products with biomimetic self-cleaning surfaces such as facade paints, polymers or textiles are already introduced. However the contribution of structures of different length scales on adhesion in various aspects still have a great potential for development and further technical applications. In addition the polymer exhibits promising features that might have impact on the development of future polymers. These features include the ability to grow, to adapt to environmental requirements and to adjust mechanical properties and will be highlighted on selected examples.
Strigolactone mixtures determine host specificity of root parasitic weeds
Yoneyama, K 1 , Xie, X 1 , Yoneyama, K 2
2
1 Weed Science Center, Utsunomiya University, Japan;
Dept of Biosciences, Teikyo University, Japan
Strigolactones (SLs) were originally isolated from plant root exudates as germination stimulants for root parasitic plants of the family Orobanchaceae, including witchweeds ( Striga spp.), broomrapes ( Orobanche and
Phelipanche spp.), and Alectra spp., and so were regarded as detrimental to the producing plants. Most of these root parasites cause devastating damages to agricultural production, in particular, in developing countries, and their effective and economically feasible control measures have not yet been established. These
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root parasites have evolved special strategies to ensure their survival; the seeds germinate only when they are within the host rhizosphere so that after germination the seedlings can reach and parasitize the host roots. To locate host roots, root parasites exploit SLs that are released by host plants for symbiotic arbuscular mycorrhizal (AM) fungi. In addition to these functions in the rhizosphere, SLs act as a novel class of plant hormones regulating shoot branching. Among the weedy root parasites, some of them attack various plant species, but the others parasitize only their specific hosts. Such specificity can be observed in germination stimulation by
SLs. For example, seeds of S. hermonthica can be induced to germinate by most of SLs including the synthetic analogue GR24, but GR24 cannot elicit seed germination of S. gesnerioide s. Furthermore, when parasite seeds are treated with a mixture of SLs, additive, synergistic, and even antagonistic effects are observed in seed germination stimulation. Since plants release their own mixtures of SLs, some root parasitic weeds with strict host specificity appear to detect quantitative and qualitative differences of SLs in the mixtures. By contrast, AM fungi may be able to reach their hosts by detecting any mixtures of SLs, as they can establish symbiotic relations with more than 80% of land plants.
Strigolactones and plant development
Brewer, PB 1
Beveridge, C 1
, Dunn, E 1 , Mason, M 1 , Meyers, E 1 ,
1 The University of Queensland, School of Biological
Sciences, St Lucia, Qld, Australia
One of the incredible features of plants is their ability to completely change their shape and structure according to environmental conditions. The ability for plants to change their fate is due to persistent meristems. These meristems are controlled by long-distance hormones that enable communication and homeostasis within the plant.
The recently discovered plant hormone, strigolactone affects meristematic processes including shoot branching, adventitious rooting and lateral root development. In addition to identifying new genetic tools aiming to tease out the biosynthetic and response pathway for this new plant hormone, our research has now expanded to understanding how this signal can regulate diverse aspects of development and is testing the hypothesis that strigolactone plays a fundamental role in diverse suppressed meristems.
Strigolactones regulate protonema branching and act as a quorum sensing-like signal in the moss
Physcomitrella patens
Proust, H 1 , Hoffmann, B 1
Yoneyama, K 2 , Nogué, F 1
, Xie, X
, Rameau, C 1
2 , Schaefer, D 3 ,
1 Institut Jean-Pierre Bourgin, UMR1318 INRA-
AgroParisTech, Centre De Versailles-Grignon, France;
2 Weed Science Centre, Utsunomiya University,
Utsunomiya, Japan; 3 Institute of Biology, Laboratory of
Cell and Molecular Biology, University of Neuchatel,
Switzerland
Strigolactones are a novel class of plant hormones controlling shoot branching in seed plants. They also
146 signal host root proximity during symbiotic and parasitic interactions. To gain a better understanding of the origin of strigolactone functions, we characterised a moss mutant strongly affected in strigolactone biosynthesis following deletion of the CAROTENOID CLEAVAGE
DIOXYGENASE 8 (CCD8) gene. Here we show that wild type P. patens produces and releases strigolactones into the medium where they control branching of protonemal filaments and colony extension. We further show that Ppccd8 mutant colonies fail to sense the proximity of neighbouring colonies, which in wild type plants causes the arrest of colony extension. The mutant phenotype is rescued when grown in the proximity of wild type colonies, by exogenous supply of synthetic strigolactones or by ectopic expression of seed plant
CCD8. Thus our data demonstrate for the first time that bryophytes ( P. patens ) produce strigolactones which act as signalling factors controlling developmental and potentially ecophysiological processes. We propose that, in Physcomitrella patens , strigolactones are reminiscent of quorum-sensing molecules used by bacteria to communicate with one another
Karrikin signalling, interaction with light and relationships with strigolactones
Nelson, DC 1
Dixon, KW 1
, Flematti, GR 1 , Scaffidi, A 1
, Ghisalberti, EL 1 , Smith, SM 1
, Waters, M 1 ,
1 University of Western Australia, Australia
Karrikins are a new family of naturally-occurring plant growth regulators discovered in smoke from wildfires.
They were discovered for their ability to break seed dormancy and stimulate seed germination. While they are active upon species that respond to fire, they are also active in species not normally associated with fire, implying a broader significance. Arabidopsis thaliana responds sensitively to karrikins, providing a means to investigate their mode of action. Recent research has shown that karrikins can change the sensitivity of seeds and seedlings to light, and influence seedling photomorphogenesis. Karrikin-insensitive mutants have been isolated and characterised, leading to the discovery that karrikin action depends on the F-box protein
MAX2/ORE9/PPS. This is the same protein that is required for response to strigolactones. Karrikins have some structural similarity with strigolactones, which are known to stimulate seed germination in some parasitic plants. However, whereas strigolactones interact with auxins to control lateral shoot development, karrikins do not have this activity. Therefore, while karrikins and strigolactones both act through MAX2, the plant can distinguish these two signalling molecules. The transport, metabolism and mode of action of karrikins will be discussed.
Can signalling crosstalk happen within the receptor molecule?
Irving, HR 1 , Kwezi, L, Donaldson, L
Wheeler, JI
Gehring, C 3
1 , Wang, YH 1
3 , Ruzvidzo, O 4
, Freihat, L 1 , Meier, S 3
1 Monash University , Australia;
Western Cape, South Africa; 3
2 University of the
King Abdullah University
,
,

of Science and Technology, Saudi Arabia;
University, South Africa
4 North West
Guanosine 3’5’-cyclic monophosphate (cGMP) has been identified as an important molecule involved in regulating a wide variety of physiological effects ranging from plant hormone dependent responses to induction of plant defence responses although the guanylate cyclase
(GC) enzymes responsible for cGMP synthesis remain elusive. We hypothesised that at least one, if not several different, functional GCs exist in higher plants.
Consequently, we designed and tested a search motif based on several functionally assigned amino acids in the catalytic domain of known GCs from lower eukaryotes and animals. The search returned over 40 putative GCs, the majority of which are annotated as receptor-like kinases (RLKs). Surprisingly, the GC catalytic centre is embedded in the kinase domain which is distinct from animal GCs where the two domains are separated. The large family of RLKs with both GC and kinase domains implies that these dual functionalities have coevolved due to the importance of both enzyme activities in plant development and stress response. These include several
RLKs known for their role in plant development and stress responses such as BRASSINOSTEROID
INSENSITIVE 1 (BRI1), WALL ASSOCIATED
KINASE LIKE 10 (WAKL10), PHYTOSULFOKINE
RECEPTOR 1 (PSKR1) and PEP1 RECEPTOR 1
(PEPR1). We have shown that these RLKs contain both kinase and GC activity in vitro. Moreover, the natural ligands of BRI1 and PSKR1 stimulate increases in cGMP in planta. Hence it is possible that the plant receptors could switch between downstream cGMP-mediated or kinase-mediated signaling cascades to elicit desired outputs to particular stimuli. The challenge now lies in understanding the interaction between the GC and kinase domains at the molecular level and how these receptors capitalize on their dual functionality in planta.
A spatio-temporal sequence of cross-regulatory events in root meristem growth
Hardtke, C
Santuari, L 1
1 , Scacchi, E
, Ragni, L
1 , Salinas, P
1
1 , Gujas, B 1 ,
1 University of Lausanne, Switzerland
A central question in developmental biology is how multi-cellular organisms coordinate cell division and differentiation to determine organ size. In Arabidopsis roots, this balance is controlled by cytokinin-induced expression of SHORT HYPOCOTYL 2 (SHY2) in the so-called transition zone of the meristem, where SHY2 negatively regulates auxin response factors (ARFs) by protein-protein interaction. The resulting downregulation of PIN-FORMED (PIN) auxin efflux carriers is considered the key event in promoting differentiation of meristematic cells. Here we show that this regulation involves additional, intermediary factors and is spatiotemporally constrained. We found that the described cytokinin-auxin crosstalk antagonizes BREVIS RADIX
(BRX) activity in the developing protophloem. BRX is an auxin-responsive target of the prototypical ARF
MONOPTEROS (MP), a key promoter of vascular development, and transiently enhances PIN3 expression to promote meristem growth in young roots. At later stages, cytokinin-induction of SHY2 in the vascular transition zone restricts BRX expression to downregulate PIN3 and thus limit meristem growth. A novel network modeling approach extending the SQUAD software that takes into account experimentally identified regulatory constraints predicted a direct involvement of
BRX in auxin signal transduction. Indeed, in vitro interaction and yeast two hybrid assays confirmed that
BRX directly interacts with MP. Interestingly, proper
SHY2 expression requires BRX, which could reflect feedback on the auxin-responsiveness of SHY2 because
BRX protein could act as a co-factor of MP. Thus, crossregulatory antagonism between BRX and SHY2 could determine ARF activity in the protophloem. Our data suggest a model in which the regulatory interactions favor BRX expression in the early, proximal meristem, while SHY2 prevails because of supplementary cytokinin-induction in the later, distal meristem. The complex equilibrium of this regulatory module might represent a universal switch in the transition towards differentiation in various developmental contexts.
Molecular and systems elucidation thermoregulation in plants
, Kakizaki, Y 1 , Elliott, C 2 , Albury, M of
2
1
Ito, K 1 , Ogata, T
Moore, A 2
1 ,
Cryobiofrontier Research Centre, Faculty of
Agriculture, Iwate University, Japan; 2 Dept of
Biochemistry & Molecular Biology, School of Life
Sciences, University of Sussex, UK
Heat production in thermogenic plants has been attributed to a large increase in the expression of the alternative oxidase (AOX). AOX acts as alternative terminal oxidase in the electron transfer chain, reducing molecular oxygen to water. However, in contrast to the mitochondrial terminal oxidase, cytochrome c oxidase,
AOX is non-protonmotive which allows the dramatic drop in free energy between ubiquinol and oxygen to be dissipated as heat. In addition to AOX, cDNAs for uncoupling protein (UCP) have been isolated for several plants, and although some tissues have been shown to coexpress UCPs with AOX, their role in promoting thermogenesis still remains unclear. In the present study, we identified cDNAs for AOX and UCP and examined their mRNA expression profiles in Arum maculatum .
RT-PCR based cloning using RNAs from the appendices of intact thermogenic inflorescences revealed that at least six cDNAs for AOX (AmAOX1a, 1b, 1c, 1d, 1e, and 1f) and one for UCP (AmUCPa) are co-expressed, but organ and developmental regulation is evident. In particular, the expression of AmAOX1e appears to be dominant in thermogenic appendices among six AmAOXs, while the expression level of AmUCPa was ubiquitous in the various tissues examined. Interestingly, predicted amino acid sequences of AmAOX1e shows that the E/DNV element found in almost all other AOX sequences, including AmAOX1a, 1b, 1c, 1d and 1f, was substituted by QNT. This is in contrast with SgAOX from thermogenic species S. guttatum in which E/DNV element is present as QDC. Further functional analyses with mitochondria isolated from yeast heterologous
147

expression system revealed that AmAOX1e is less sensitive to stimulation by pyruvate. These data, together with our comparative studies on temperature and proteomic analyses of A. maculatum and S. renifolius , are interpreted in terms of the possible mechanism for thermoregulation in plants.
Alternative oxidases of non-angiosperm plants
McDonald, A 1
1 Wilfrid Laurier University, Waterloo, Canada
Biological electron transport chains (ETCs) power life on this planet. In most eukaryotic organisms, ETCs in mitochondria generate energy via oxidative phosphorylation. This ‘classic’ ETC is present in mammals, but it does not accurately reflect respiration in most prokaryotes and eukaryotes. Typically, linear ETCs are depicted in textbooks, however, this is misleading, as most organisms have additional protein complexes that increase the points of entry and/or exit of electrons, resulting in a branched chain. Alternative oxidase (AOX) is a terminal ubiquinol oxidase present in mitochondrial electron transport chains that introduces a branch-point in respiration and can bypass two of the three proton pumping complexes (Complexes III and IV). This leads to a decrease in the amount of ATP that can potentially be generated by ATP synthase. Plant AOX research to date has focused on angiosperms such as Arabidopsis thaliana , tobacco, soybean, and rice, where a good deal is known about its multigene family, gene expression, and the post-translational regulation of AOX activity. In contrast, little is known about AOX in other plant groups.
We utilized a bioinformatics approach to investigate the taxonomic distribution of AOX in non-angiosperm members of the Viridiplantae (green plants) and used multiple sequence alignments of AOX proteins to investigate the distribution of the AOX multigene family, the AOX1 and AOX2 subtypes, and amino acid residues involved in the catalytic function and post-translational regulation of the enzyme. AOX sequences were found in many members of the Viridiplantae, in both
Chlorophytes and Streptophytes, including liverworts, a moss, a lycopod, ferns, and several gymnosperms. We are employing reverse transcriptase PCR using degenerate primers as a strategy to amplify novel AOX sequences from plant phyla for which molecular data are not yet available (e.g. Psilotophyta, Equisetophyta,
Ginkophyta, and Cycadophyta). A comparison of AOX sequences at the amino acid level from angiosperms and non-angiosperms reveals similarities and differences which have direct implications for the evolutionary history of AOX, AOX catalytic function, and the posttranslational mechanisms that regulate AOX activity in plants.
Elucidating the mechanism of thermogenesis in
Philodendron bipinnatifidum
– elevated oxygen concentrations overcome diffusional limitations to isotope discrimination during respiration in dense thermogenic tissues
Miller, R 1,2,3 , Grant, N 2
Berry, J 4 , Watling, J 3
, Giles, L
, Robinson, S 2
4 , Ribas-Carbo, M 5 ,
1 School of Biological Sciences, Monash University,
Clayton, Australia; 2 Institute for Conservation Biology and Environmental Management, The University of
Wollongong, Wollongong, Australia; 3 Ecology &
Evolutionary Biology, School of Earth and
Environmental Sciences, The University of Adelaide,
Adelaide, Australia; 4 Dept of Global Ecology, Carnegie
Institution of Washington, Stanford, USA; 5 Universitat de les Illes Balears, Departament de Biologia, Unitat de
Fisiologia Vegetal, Illes Balears, Spain
The thermogenic inflorescences of Philodendron bipinnatifidum heat up to 34ºC above ambient temperature by achieving high mass specific respiratory rates, equivalent to those in some animals, yet the mechanism of heating in this and other arums has not been definitively characterised. Transcripts for both the alternative oxidase (AOX) and plant uncoupling protein
(pUCP) – the two possible pathways for heat generation in plants – have been found in thermogenic tissues of P. bipinnatifidum . The only means to definitively demonstrate the mechanism of heat production in vivo is to quantify respiratory pathway fluxes using stable O
2 isotope discrimination techniques; however these measurements have not been possible in thermogenic
Araceae to date because of the high diffusional resistances in these structurally dense tissues. We aimed to investigate the contribution of the alternative oxidase
(AOX) and uncoupling proteins (pUCPs) to heating, in both fertile (FM) and sterile (SM) male florets of P. bipinnatifidum using oxygen isotope discrimination.
Specifically we aimed to investigate whether isotopic discrimination was affected by diffusion during peak respiration in SM florets, by conducting measurements under different O
2
partial pressures. We also characterised heating patterns and mechanisms in the little studied fertile male (FM) florets. We found that FM florets heat independently of SM florets, and both heat for up to 30 hours after removal from the plant. In FM florets, heat production was strongly correlated with both total respiration and AOX pathway flux. In contrast, whilst heating was also strongly correlated with total respiration in SM florets, AOX flux was not. To determine whether this was a function of pUCP activity or an artefact of diffusional limitations in SM tissues, we measured isotope discrimination under elevated O
2 partial pressures in the most strongly heating tissues.
These measurements confirmed that diffusional limitations were responsible for the low AOX fluxes observed in SM florets in air. Under increased O
2
, AOX contributed up to 92% of total flux in peak heating SM florets. Elevated O
2
can thus be used to measure respiratory pathway flux in dense tissues. Protein data support the substantial role for AOX in thermogenesis in
P. bipinnatifidum . Both pUCP and AOX proteins were present in thermogenic tissues; however, AOX protein increased 5-fold with the onset of thermogenesis in both floret types whereas pUCP remained low throughout development. No relationship between AOX protein content and heating was found, suggesting that fine scale regulation of AOX is post-translational.
Flux distributions within the heterotrophic plant
TCA cycle
Ratcliffe, RG
SK 1
1 , Beard, KFM 1
, Sweetlove, JL 1
, Kruger, NJ
, Williams, TCR 2
1 , Masakapalli,
148

1 University of Oxford, Oxford, UK;
Federal de Vicosa, Vicosa, Brazil
2 Iniversidade
The input of acetyl CoA into the tricarboxylic acid
(TCA) cycle allows the cycle to maintain a cyclic flux that lies at the heart of mitochondrial respiratory metabolism. This flux supports the oxidation of a series of carboxylic acids, generating the reducing equivalents –
NADH and FADH2 – that fuel the synthesis of ATP by oxidative phosphorylation. The TCA cycle is embedded in a wider metabolic network, allowing it to support both cyclic and non-cyclic flux modes, and the observed flux distribution is dependent on the metabolic and physiological demands of the cell (1). Labelling experiments and metabolic modelling provide the key to analysing the flux distribution in the TCA cycle, and several groups have harnessed the power of steady-state metabolic flux analysis (MFA) to analyse the cycle in heterotrophic tissues. Steady-state MFA is used to generate flux maps of the core network of central carbon metabolism and the TCA cycle fluxes are often well defined (2). Flux maps are based on observations of the redistribution of an isotope, typically 13C, in labelling experiments and robust protocols have been developed to allow the method to be applied to the extensively compartmented network of heterotrophic metabolism in plant cells. Applications to Arabidopsis cell cultures have revealed physiologically important features of the network, such as its robustness (3) and the balance between cytosolic and plastidic metabolism (4).
Moreover a comparison between MFA and constraintsbased stoichiometric modeling has shown that the latter approach is capable of accurately predicting some of the responses of central metabolism to the imposition of stress conditions (5). The results of several steady-state
MFA analyses of the TCA cycle in a heterotrophic
Arabdiopsis cell culture will be discussed, including current investigations of the impact of phosphorus and nitrogen nutrition on central carbon metabolism and carbon use efficiency. The extent to which flux maps capture the expected features of highly compartmented metabolic networks is variable, and a further complication that has rarely been considered, and which might be significant for the TCA cycle, is the potential existence of metabolite channelling. This process can have a significant impact on the redistribution of isotopic label – indeed evidence for channelling is frequently obtained from isotope dilution experiments – but this aspect of metabolism has been largely ignored in the development of steady-state MFA for plants. Theoretical analysis of simple networks, and re-analysis of GC-
MS/NMR datasets obtained for Arabidopsis cell cultures, suggests that the effects of channelling are detectable in parts of the network and in parallel with this in silico approach, isotope dilution experiments are being conducted on isolated mitochondria to test the extent to which channelling occurs between intermediates in the
TCA cycle. (1) L.J. Sweetlove et al. 2010. (2) N.J.
Kruger & R.G. Ratcliffe 2009. (3) T.C.R. Williams et al.
2008. (4) S.K. Masakapalli et al. 2010. (5) T.C.R.
Williams et al. 2010.
Gene expression and metabolite signatures associated with global change treatments that engage or disengage mechanisms maintaining homeostasis between respiration and photosynthesis
Leakey, ADB
EA 1
1 , Richter, K 1 , Gillespie, KM 1 , Ainsworth,
1 University of Illinois At Urbana–Champaign, USA
Understanding and modeling of plant and ecosystem carbon balance has long been challenged by uncertainty about how and when homeostasis is maintained between photosynthetic and respiratory fluxes. We present a case study of how gene expression and metabolite signatures can reveal the metabolic and biosynthetic responses in leaves distinguishing: (a) enhanced dark respiration coupled with greater photosynthesis and productivity at elevated [CO ] from (b) enhanced dark respiration
2 coupled with impaired photosynthesis and productivity at elevated [O
3
]. Soybean was grown over its entire lifecycle under ambient atmospheric conditions, elevated
[CO
2
] and elevated [O
3
] treatments using free-air concentration enrichment (FACE) at a field site in the
Midwest U.S.. Stimulated foliar respiration at night under elevated [CO
2
] was associated with enhanced photosynthetic carbon gain, larger pools of sugar and starch substrates, and greater abundance of transcripts encoding the respiratory machinery. This is consistent with transcriptional reprogramming of metabolism to increase respiratory capacity in response to: (1) greater substrate availability from photosynthesis and, (2) greater demand for energy from phloem loading associated with additional photoassimilate export to support enhanced whole-plant growth. Stimulated foliar respiration at night under elevated [O ] was associated with reduced
3 photosynthetic carbon gain, but greater antioxidant capacity, ascorbate content and greater abundance of transcripts encoding respiratory, anti-oxidant and chlorophyll synthesis pathways. This is consistent with transcriptional reprogramming of metabolism in response to the necessity for energy and carbon skeletons from respiration to support detoxification and repair processes associated with oxidative stress, despite reduced substrate availability. The decoupling of respiration from photosynthesis under elevated [O
3
] greatly exacerbated the negative consequences for carbon balance at the leaf and whole-plant scales. These results suggest that both sugar status and stress signals can drive modulation of respiratory capacity in leaves. Further analysis of transcription factors and other regulatory elements displaying altered gene expression in these experiments presents an opportunity to determine how sugar and stress signals are integrated to engage or disengage mechanisms maintaining homeostasis between respiration and photosynthesis.
Unraveling the acclimation of the mitochondrial proteome to changing environments
1
Taylor, N 1,2
Tan, Y-F
, Huang, S
2 , Millar, H 2
2 , Jacoby, R 2 , Shingaki-Wells, R 2 ,
ARC Centre of Excellence In Plant Energy Biology ,
Australia; 2 Centre for Comparative Analysis of
Biomolecular Networks , Australia
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Changes in the environment in which a plant grows can have dramatic affects on the metabolic processes of photosynthesis and respiration. This in turn can have major implications on biosynthesis, cellular maintenance and biomass allocation. Analysing the network of changes under different environments is essential to understand the plasticity of this acclimation system as a whole. Here we investigate three examples of the effects of a changing environment on mitochondrial function and protein content. 1. Plants can acclimate to the extremes of temperature following a pre-exposure to a lower sub-lethal increase/decrease in temperature that allows them to adjust their metabolism and to survive.
The respiratory and photosynthetic rates of these plants remain similar to plants grown at optimal temperatures and plants that have a greater thermal tolerance tend to more quickly adjust their metabolism and restore respiration to pre-thermal change levels. Using both
Arabidopsis whole plants and cell culture we have produced data that suggests a remodeling of proteins in the mitochondrial electron transfer chain allows respiratory homeostasis to be achieved during acclimation. Also changes in other mitochondrial membrane and soluble proteins have been measured quantitatively at the proteome level following chilling. 2.
While rice is cultivated on flooded/anaerobic soils many other dry-land crop species suffer when encountering waterlogging leading to O
2
deficiency. Rice is an ideal model species with an available genome sequence for elucidating the mechanisms of anoxia tolerance in plants and because it can survive under prolonged anoxia.
Without O
2
, the glycolytic pathway that is linked with ethanolic fermentation is the predominant mechanism of energy production in plants. However, much less energy is produced during anoxia and the synthesis rate of proteins will decrease well below that of what is seen in aerated tissues. Even so, rice coleoptiles still exhibit a complex pattern of newly synthesized proteins. Along with classical anaerobic proteins, anoxic rice coleoptiles also synthesize some proteins with unknown functions and mitochondria initiate specific anaerobic metabolic functions. 3. Salinity is a severe impediment to agricultural productivity and with the increased use of marginal land, breeders will need to produce crop and fodder plants that exhibit high salinity tolerance. Salinity is an acute problem in the Australian wheat growing regions. We have found quantitative differences in the shoot mitochondrial proteomes of v.Wyalkatchem and v.Janz, two commercially important wheat varieties that are known from a range of experiments to have differing salinity tolerance. These proteins included Mnsuperoxide dismutase (Mn-SOD) and the alternative oxidase (AOX), both of which have previously linked to reduced ROS formation from the electron transport chain and salt tolerance in Arabidopsis . Together, the data presented here suggest that changes in mitochondrial
ROS defense pathways in the mitochondrial proteomes of key Australian wheat varieties correlate with wholeplant salinity tolerance. As plant biomass accumulation is governed by the equation of assimilation of CO
2 respiratory CO
2
minus
loss, the rate, degree and timing of changes in respiratory function is a critical component in plant growth and provides an insight to the dynamic nature of the mitochondrial proteome.
Fern venation as a possible evolution from tree-like to net-like vein architecture
1
Douady, S 1
MSC Laboratory, CNRS-Paris Diderot University,
France
Following the evolution, the leaf venation has come roughly from dichotomous tree-like for the ferns, to reticulate in the dicotyledons, back to parallel veins with small cross connections in the monocotyledons. When looked at the tree-like pattern of the first ferns, it is difficult to understand how such a dramatic change in pattern could have led to the net-like fully reticulated patterns of the dicotyledons. However, a detailed study of the pattern of veins in neotropical ferns can reveal a complete variety of patterns, from the full dichotomous tree-like pattern, to the full reticulation, even in a simple genus such as Adiantum , or Thelypteris . It can indeed be observed the first connection of only two veins at he bottom of two leaflets, to the common connection of all the third order veins at the middle line, creating an intermediary line and a continuous leaf, to more intermediary connections to finally the full reticulation.
These intermediary patterns are particularly interesting as they could reveal how and why the transition happened.
All these patterns can be analyzed with tools developed for studying other reticulate patterns like porcelain cracks and street network. It is interesting to classify all these patterns, to see if it is possible to organize them along a single line of evolution, to see if there is a unique path, and to compare the result with what is know from the evolution of these ferns.
The evolution of maple leaf size, shape, composition,
1 stomata and venation architecture
Sack, L 1,2 , Scoffoni, C 2 , Barron, R
Corresponding Author; 2
3 , Frole, K 4 , Zanne, A 5
University of California, Los
Angeles, Dept of Ecology and Evolutionary Biology,
USA; 3 Harvard University, Dept of Organismic and
Evolutionary Biology, Biological Laboratories,
Cambridge, USA; 4 University of Hawaii, Botany Dept,
Honolulu, USA; 5 University of Missouri St Louis,
Biology Dept, St Louis, USA
Species of maple ( Acer , Sapindaceae) are highly variable in leaf form, e.g., ranging from compound to simple leaves and in shape from entire to 11 lobes. We determined the variation, phylogenetic signal and evolutionary coordination of over 50 traits relating to size, shape, nutrient composition, venation architecture and stomatal anatomy, for well-illuminated leaves of 30 maple tree species of North America, Europe and Asia,
150

grown in a common garden (Arnold Arboretum of
Harvard University). We also determined intra-canopy plasticity for key leaf traits. We comprehensively tested expectations for how traits should correlate with shadetolerance and drought tolerance based on previous studies of species variation within and across communities. We found evidence of strong traitenvironment linkages, but that these held only in the minority of expected cases, indicating that trait adaptation to habitat is not direct or predictable in the simplest ways, and we must consider simultaneous adaptation to multiple environmental axes, and evolution of multi-trait clusters rather than single traits. Indeed, we found novel, tight relationships among traits within clusters related to leaf size, or shape, or composition, or venation, or stomatal anatomy, and also within clusters as defined by function (e.g., flux-related traits and leaf economics traits). However, we found impressive evolutionary independence among these trait clusters.
The independence of leaf trait clusters results in the possibility of many trait configurations in leaf form even among species similar in habitat and successional stage
(e.g., pioneer or shade-establishing). Across the maple species, we observed strong variation among traits in their phylogenetic signal across the lineage. Across the traits considered, trait phylogenetic lability was independent of trait diversification (i.e., the degree the trait varied across extant species). Thus, strong trait diversification can arise equally frequently from multiple trait shifts among closely-related species as from few shifts deeply nested in the tree, and trait similarity across the genus can arise equally from phylogenetic conservatism as from frequent shifts within narrow bounds. Such multiple pathways to trait similarity and divergence highlight how evolutionary flexibility potentiated the rich total diversity in maple leaf form.
Finally, across the traits we found a correlation of a trait's developmental plasticity within the canopies of individual trees with both its phylogenetic lability and diversification across species in the genus. This work provides insights into the genetic basis for leaf traits, and highlights the necessity for mechanistic understanding of how multiple leaf traits contribute to integrated function, during adaptation within and across habitats. Key words: leaf traits, stomata, scaling, phylogeny, venation architecture, leaf morphology.
Leaf hydraulic vulnerability is related to conduit dimensions and drought resistance across a diverse range of woody angiosperms
Blackman, C 1 , Brodribb, T 1 , Jordan, G 1
1 School of Plant Science, University of Tasmania,
Australia
Hydraulic dysfunction in leaves determines key aspects of whole-plant responses to water stress, however, our understanding of the physiology of hydraulic dysfunction and its relationships to leaf structure and ecological strategy remain incomplete. Here, we studied a morphologically and ecologically diverse sample of angiosperms to test whether the water potential inducing a 50% loss in leaf hydraulic conductance (P50leaf) is predicted by properties of leaf xylem relating to water tension-induced conduit collapse. We also assessed the relationships between P50leaf and other traits considered to reflect drought resistance and ecological strategy.
Across species, P50leaf was strongly correlated with a theoretical predictor of vulnerability to cell-collapse in minor veins (the cubed ratio of conduit wall thickness to conduit lumen breadth). P50leaf was also correlated with mesophyll traits known to be related to drought resistance, but unrelated to traits associated with carboneconomy. Our data indicate a link between the structural mechanics of leaf xylem and hydraulic function under water stress. Although it is possible that collapse may directly contribute to dysfunction, this relationship may also be a secondary product of vascular economics, suggesting that leaf xylem is dimensioned to avoid wall collapse.
Decline of leaf hydraulic conductance during desiccation: relationships to leaf size and venation architecture in species varying strongly in drought tolerance
Scoffoni, C 1 , Rawls, M 1 , McKown, A 1 , Sack, L 1
1 University of California, Los Angeles, USA
Leaves vary greatly in their size and venation architecture. Leaf veins deliver water to replace that lost to transpiration, allowing stomata to remain open for photosynthesis. Previous studies have shown that the leaf hydraulic conductance (Kleaf) is influenced by venation architecture. Severing the midrib caused a decline in
Kleaf and gas exchange to a degree that depended on venation architecture: leaves of higher major vein density
(length/area) had greater tolerance of hydraulic disruption due to additional pathways for water flow around the severed vein. Because desiccation-induced vein embolism also reduces Kleaf, we hypothesized that leaves with higher major vein density and thus greater redundancy of flow paths would have a lower hydraulic vulnerability. Further, smaller leaves, which tend to have intrinsically higher major vein densities, should tend to have lower hydraulic vulnerability. For ten species ranging strongly in drought sensitivity, we determined the relationships among leaf hydraulic responses to desiccation and rehydration, venation architecture, gross morphology, and other drought tolerance traits. We quantified Kleaf using the evaporative flux method for leaves of a range of water potentials (Øleaf). Hydraulic vulnerability, calculated as the Øleaf at 50% and 80% loss of Kleaf, correlated negatively with major vein density and positively with leaf size (|r |= 0.85 to 0.93; P
= 0.01-0.002). These relationships were independent of other aspects of drought tolerance. Hydraulic vulnerability was independent of minor vein density, consistent with computer simulations showing the importance of the hierarchical arrangement of veins.
These findings suggest a new functional importance of high major vein density and small leaf size in drought tolerance and ecological specialization, potentially important in as a determinant of biogeographic patterns in leaf size.
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The ghosts that Darwin missed: pollination of
Caladenia
by sexually deceived wasps … and others
Petit, S 1
1 University of South Australia, Australia
The complexity of Caladenia (syn. Arachnorchis ) species’ reproductive success remains shrouded in mystery. I will unravel some of the mystery surrounding the pollination of spider orchids (leaving plenty for others to unravel). By mimicking the pheromone of a wingless female insect, an orchid tricks the sexually aroused winged male. Are the pollinators simply silly thynnid wasps? What makes an effective pollinator?
What makes an orchid a successful temptress?
Pollinators are difficult to study in species with relatively low pollination rates, but their ghostly behaviours can be derived from examining plant characteristics in relation to pollination success. The orchid masters the art of treachery, but will it be its demise?
Pollination systems of Angraecoid orchids: from
Darwin to the present day
Micheneau, C 1, 2 , Fournel, J 2
Gauvin-Bialecki, A
M 4
2
, Warren, B 2 , Hugel, S 3
, Pailler, T 2 , Strasberg, D 2
,
, Chase,
2
3
1 Université Libre de Bruxelles, Brussels, Belgium;
Université de La Réunion, Saint-Denis, La Réunion;
4
CNRS – Université de Strasbourg, Strasbourg, France;
Royal Botanic Gardens, Kew, Richmond, UK
Since the famous prediction of Darwin, the spectacularly long nectar spurs of Madagascan Angraecum orchids have represented a classic example of floral adaptation to long-tongued hawkmoth pollination. Although specialisations for a specific group of pollinators can be viewed as a key innovation that has been responsible for the evolutionary success of a lineage, extreme specialisations in turn can be linked with loss of evolutionary potential, decreasing the ability of species to adapt to future environmental change. Here we report a surprising pollinator shift in the orchid genus Angraecum from hawkmoth to bird and raspy-cricket pollination (in the insect order Orthoptera). This evolutionary shift occurred after this group of orchids dispersed to a new insular habitat that harboured both different environmental conditions and groups of pollinators than were present in their native island, Madagascar. This specialised plant-pollinator interaction further represents the first clear case of orthopteran-mediated pollination in flowering plants.
Total evidence phylogenetic analysis of the orchid tribe Diurideae and what it tells us about the evolution of pollination systems
Weston, PH 1
Peakall, R 4
, Indsto, JO 2 , Perkins, AJ 1 , Clements, MA 3 ,
1 National Herbarium of New South Wales , Australia; 2
Loftus Road, Pennant Hills, Australia;
Industry , Australia;
Australian National University , Australia
3
34
Centre for
Australian National Biodiversity Research, CSIRO Plant
4 Research School of Biology, The
The tribe Diurideae is a clade of terrestrial orchids that has its centre of diversity in Australia, but some of its sub-clades extend into the south-western Pacific and southern and eastern Asia. Most species are perennial deciduous herbs that die back to dormant tubers at the beginning of summer, renewing active growth the following autumn. Annual growth in most species involves the replacement of the old tuber with a single new one and growth of a single-leaved shoot subtending a multi-flowered raceme. The Diurideae includes taxa that offer nectar to their pollinators (e.g. Prasophyllum ) as well as those that lure pollinators by mimicking: 1. food sources (e.g. some species of Diuris ) or 2. female hymenopteran insects (e.g. Drakaeinae) or 3. brood-sites of female flies (e.g. Corybas ) and it also includes a diverse assemblage of self-pollinating and apomictic taxa. The fruits of the underground orchids, Rhizanthella , are berries enclosing large, crustose, presumably animaldispersed seeds but those of all other taxa are capsules that release dust-like, wind-dispersed seeds. The phylogeny of the orchid tribe Diurideae will be reconstructed down to the level of species groups using a combined matrix of morphological and anatomical characters and alignments for 4 genes ( mat K, trn L-F, rbc L, psb A), plus the intergenic spacer ITS. The data will be phylogenetically analysed using parsimony and
Bayesian methods. The molecular partition of the dataset will also be subjected to molecular dating analyses using
Bayesian relaxed clock methods, calibrated using a recently described orchid fossil as well as secondary calibrations based on non-orchid monocot fossils and the known ages of some biogeographic disjunctions.
Preliminary analyses suggest the following:
• The crown group of Diurideae diversified in Australia after final separation of the continent from Antarctica about 35 mya and several lineages subsequently dispersed to south western Pacific islands and into Asia across ocean gaps
• Deciduousness, annual production of a single replacement tuber, a multi-flowered raceme and capsular fruit are ancestral character states relative to evergreen habit, absence of tubers, production of multiple replacement tubers, single-flowered inflorescence and baccate fruit
• Food source mimicry is ancestral relative to both sexual mimicry and nectar production
• Sexual deception of male Hymenoptera has evolved independently in at least 6 different lineages
( Cryptostylis, Calochilus, Leporella, Drakaeinae, at least two distinct clades in Caladenia )
• Nectar production has evolved multiple times (in
Prasophyllinae, Acianthinae, Diuris, Eriochilus ,
Caladenia )
• Several clades of closely related species include both nectar-producing and nectarless species ( Diuris,
Caladenia, Eriochilus ) and multiple, possibly bidirectional transformations between these states have occurred in these clades
• Autogamous and apomictic lineages have evolved numerous times from food source mimicking, sexually deceptive and nectar-producing ancestors What selective
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pressures have facilitated the evolution of sexually deceptive lineages from food-source-mimicking ancestors? Pollinator reliability seems the best answer.
Explaining the evolution of nectar-producing lineages from food-source-mimicking ancestors and vice versa seems a much more complex problem. Species groups such as Diuris section Purpureo-albae that include both nectar-producing species and food-source mimics offer useful model systems for exploring this problem.
Pollinator-driven evolution of orchids in southern
Africa
Johnson, S 1
1 University of KwaZulu-Natal, South Africa
The orchid diversity in southern Africa is largely the outcome of radiation in a single terrestrial clade. There is now overwhelming evidence that much of the diversification of the larger genera in this clade, such as
Disa and Satyrium , is the consequence of evolutionary shifts between different pollinators. Orchids generally form part of broader pollination guilds, and frequently exploit other guild members through floral mimicry. This is particularly evident in Disa where many members of non-rewarding clades have been identified as Batesian food mimics. Species-level phylogenies are now available for most of the larger orchid genera in southern
Africa, allowing both the direction and frequency of pollinator shifts to be estimated. In addition, these phylogenies allow the degree of lability and covariation in key floral traits, such as nectar, colour, shape and scent, to be assessed for the first time.
Orchid colour and pollination by deception
Gaskett, AC 1
1 The University of Auckland, New Zealand
Visual signals such as floral colour are primary pollinator attractants. Orchids have long been thought to use colour in pollinator attraction and deception, but colour is likely to function quite differently in rewarding than in nectarless or deceptive pollination systems. Darwin supported Sprengel's then-controversial hypothesis that showy floral colours were pollinator attractants.
Consequently, Darwin noted that he had few explanations for why pollinators were attracted by orchids with drab green or inconspicuous flowers, many of which had hinged labellae. These non-traditional floral features are now strongly associated with sexually deceptive pollination systems in which orchids mimic female insects and pollination occurs when male insects are fooled into sexual behaviour with flowers. Although sexual deception primarily relies on attracting pollinators with olfactory signals, floral colour can still play a role.
Orchids may mimic the colour of female insects, or more intriguingly, exploit pollinators’ sensory biases or innate preferences for certain wavelengths and colour contrasts.
Research into the functions of colour in deceptive pollination systems has focussed largely on fooddeceptive orchid species that are nectarless but nonetheless still pollinated by foraging male and female insects. In food deceptive orchid species, colour can function in Batesian floral mimicry of specific, nearby, rewarding model flowering plant species, although convergent evolution is a potentially confounding factor.
The function of colour polymorphism is less clear in food deceptive orchid species exhibiting more generalised mimicry without a specific model species. Although some studies suggest colour polymorphism is due to frequency dependent selection by pollinators, several other abiotic and environmental explanations may be more influential. New technologies and devices provide affordable means to measure spectral wavelengths reflected by orchids, but how are these colours seen by pollinators? Competing models of insect vision mean interpreting pollinator perception of orchid spectral reflectivity remains controversial. Here I review and evaluate the most common methods of modelling spectral reflectance and trichromatic hymenopteran pollinator vision: hexagonal colour opponency (as proposed by Chittka) and triangular non-colour opponency (as proposed by Vorobyev and Brandt). I discuss suggested methods and techniques for data collection, and performing calculations manually and with software. I also present new data on floral colours of sexually deceptive and putatively brood-site deceptive orchid species from Australia and New Zealand. Broodsite deception involves mimicry of typical oviposition sites and therefore pollination by female rather than male insects. Brood-site mimicry by orchids has received less attention than other forms of deceptive pollination, although anecdotal evidence is strong. If deception imposes different costs on the reproduction of female insects than on male insects, each form of deceptive pollination may involve distinct selective pressures.
Consequently, the functions and roles of visual versus olfactory signals may vary between these pollination systems.
Impacts of extreme weather on flowering phenology of wild orchids in southwestern China
Liu, H 1 , Feng, C-L 2 , Luo, Y-B 3 , Deng, Z-H 4
2
1 Florida International University, Fairchild Tropical
Botanic Garden, USA, and Guangxi University, China;
Experimental Center of Troical Forestry, Chinese
Academy of Forestry Sciences, China; 3 State Key
Laboratory of Systematic and Evolutionary Botany,
4
Institute of Botany, Chinese Academy of Sciences, China;
Yachang National Orchid Nature Reserve, Guangxi,
China
Impacts of unusual weather on orchid reproduction was mentioned by Darwin in his 2nd edition of the book On the various contrivances by which orchids are fertilised by insects . However, rare weather events as drivers of temporal variations in reproduction of orchids were not emphasized by him or other researchers. Yet, among the predictions of on-going climate change, is increased frequency of extreme weather events. Therefore, it is of paramount importance to understand the impacts of extreme weather events on orchid reproduction. Here we explored the effects of an extreme cold event during early spring of 2008 on flowering phenology of 50 species in 21 genera of wild orchids in the Yachang
Orchid Nature Reserve in southwestern China. We found that 23 species (46%) delayed their flowering time compared to the previous year that had average spring weather, while 27 species (54%) did not delay. We
153

related these phenological responses to various ecological traits, include flowering season (spring vs. non-spring), habit (terrestrial vs. epiphytic/lithophytic), endemicity (narrow endemic vs. non-narrow endemic), population size (small vs. large population), and range position (southern-most population or non-southern-most population). We found that only population size had significant impacts on whether the species had a phenological response to the extreme low spring temperature. Specifically, 60% of the species with relatively large populations responded with a flowering delay while only 25% of the species with very small populations did. While terrestrial, spring flowering orchids and species with a wide global distribution range were more likely to delay flowering than epiphytic, summer or fall flowering, and narrow endemic orchids, these differences were not statistically significant. A comprehensive logistic regression analysis (backward stepping), which incorporated all the explanatory variables also confirmed that population size was the only significant factor. Since species that were not sensitive to environmental cues were more likely to go extinct during the past climate change events, orchids with small populations in southwestern China may be more vulnerable to the current and projected climate change than orchids with large populations due to lack of responses to environmental changes of the former group.
Biochemical evolution of pollination drops in seed plants von Aderkas, P 1
1 Graduate Centre for Forest Biology, University of
Victoria, Canada
During the evolution of seed plants, in particular from pteridosperms through gymnosperms, pollen was captured by pollination drops. This is still the case for almost all modern gymnosperms. A small number of angiosperms have homologous ovular secretions. In gymnosperms, these drops are secreted from the nucellus, filling pollen chambers and/or micropylar canals. Spheres of extruded liquid form at the outer entrance to ovules. The drops are a component of pollination mechanisms that have evolved to deliver pollen into the ovule. The drops also induce pollen germination. In species in which pollen is captured by physical means, e.g. Douglas-fir and larch, drop secretion and, consequently, pollen germination are delayed by many weeks. Although some podocarps have pollination drops that are able to scavenge for pollen, most pollination drops indiscriminately accept whichever pollen species land. Some evolutionary innovations exist, such as sacci, that improve the chances for the ovule of selecting homospecific pollen. Intergeneric crosses reveal some chemically based selective mechanisms for prezygotic seclusion of undesirable pollen. In some species, pollination drops are secreted once only, in others they are repeatedly secreted. In addition, there is some evidence that pollen capture may be followed almost immediately by drop resorption. With such a
154 diversity in behaviour and phenology, it is also not surprising that the biochemical composition should show evolutionary differences. Most drops include sugars, amino acids, a limited number of secondary compounds and calcium, as well as many proteins. Concentrations of components, in particular sugars, show broad differences associated with types of pollination. Gymnosperms include species that are wind-pollinated and/or insectpollinated. The latter share compositional similarities with nectar. Each gymnosperm pollination type has a characteristic type of pollination drop, both in terms of chemical composition and volume range. Windpollinated species tend to have lower volumes and lower concentrations of sugars than insect-pollinated species.
We have molecular biological evidence that compounds in the drops originate from the sporophyte, not the megagametophyte. Proteins have a variety of roles including alteration of carbohydrate composition (e.g. invertases, xylosidases, galactosidases), defense against microbes (e.g. chitinases, peroxidases, glucanases), and prevention of ice damage (TLPs). A number of elements are not yet understood. Work continues on surveying proteins in the gymnsoperm clade. The mechanism that regulates quick retraction of pollination drops is not known. Genomics approaches to reproduction will likely prove very rewarding in elucidating the evolution of pollination drops in seed plants.
‘Toxic nectar’: the responses of pollinators to nicotine in artificial nectar
Nicolson, S 1 , Koehler, A 1
1 University of Pretoria, South Africa
The presence of secondary metabolites (SM) in floral nectar seems paradoxical, in view of the reward function of nectar, and little is known of their role in mediating interactions between plants and pollinators. Nectar SM may have multiple ecological functions, including protection from microbial infection and unwanted visitors, and attraction of specialised pollinators. We are investigating the response of various pollinators to nicotine, as a model alkaloid, in their nectar diets.
Nicotine, best known from the Solanaceae, is one of the best-studied SM in terms of defence against herbivory. It is highly toxic to most herbivores through its action on acetylcholine receptors. Nectar is chemically diverse, and the attractive or deterrent effects of SM such as nicotine may depend on other nectar attributes. The most obvious of these is nectar concentration. Using preference tests with two very different pollinators – the whitebellied sunbird Cinnyris talatala and the honeybee Apis mellifera – we have shown that the dose-dependent deterrent effect of nicotine is stronger in more dilute nectars. Modulation of the deterrent effect of nectar alkaloids by nectar sugar concentration is consistent with
SM in nectar acting as partial deterrents that keep pollinators moving between plants and ensure crosspollination. Short visits by pollinators are not a disadvantage in terms of plant fitness. From the animal point of view, stronger deterrence in dilute nectars may be considered beneficial when compensatory feeding, as in nectar-feeding birds, potentially increases the alkaloid intake. In coping with SM in nectar, pollinators must achieve a balance between energetic gains and palatability. We have also investigated the response to

nectar nicotine of the giant day gecko Phelsuma madagascariensis grandis : these animals are attracted by
0.5 µM nicotine, repelled by 1000 µM, and indifferent to intermediate concentrations. Pollinator declines are currently of great concern internationally, and the contributing factors are thought to include pesticides such as neonicotinoids. These act at the nicotinic acetylcholine receptors, and may have adverse effects on nontarget insects and especially honeybees. We are therefore interested in the physiological consequences of ingesting nicotine in honeybees. When newly emerged workers were caged in incubators and fed varying concentrations of nicotine in sucrose solution for 21 days, consumption did not depend on nicotine levels in the diet. Survival of the bees decreased on high nicotine, but was unaffected by low and medium nicotine concentrations. However, in three weak colonies nicotine increased survival. It is possible that low levels of SM in nectar may provide protection against disease, as already shown for bumblebees. In future studies, we will investigate how sublethal levels of nectar SM affect honeybee foraging behaviour and trophallactic exchanges among bees, and how toxin levels are changed by the processing of nectar into honey. Sublethal effects of SM may be more subtle but still detrimental for pollinators.
The role of scent compounds in attraction of rodent pollinators to flowers of the South African Pineapple
Lily,
Eucomis regia
(Hyacinthaceae)
Wester, P 1, 2 , Pauw, A 2 , Johnson, S 1
1 School of Biological and Conservation Sciences,
University of KwaZulu-Natal,
Pietermaritzburg, South Africa; 2
Scottsville,
Dept of Botany and
Zoology, University of Stellenbosch, Stellenbosch, South
Africa
Plants adapted to rodent pollination show characters like visually inconspicuous, bowl-shaped flowers near ground level, stiff stamens, easily accessible nectar and often characteristic scent. The South African Pineapple lily
Eucomis regia (Hyacinthaceae) is hypothesised to be rodent-pollinated on the basis of sharing these characters and as it differs from insect-pollinated Eucomi s species mainly in scent chemistry. Under natural conditions and in the laboratory mice were observed to become dusted with pollen as they licked nectar in the flowers. Pollen and dye was transferred to stigmas. Live-trapped mice had large amounts of E. regia pollen in the fur around the snouts and in the faeces. Selective exclusion of vertebrates, but not insects, led to significant reductions in seed set. Controlled pollination experiments showed that E. regia is self-incompatible and thus entirely dependent on pollinator visits for seed production.
Spectral reflectance of floral tepals is very similar to the green bracts and leaves, rendering flowers inconspicuous to insects. The scent of flowers and nectar is reminiscent of cooked potatoes due to the presence of the sulphur compound methional. Choice experiments showed that mice are strongly attracted to this compound.
Phloem at the epidermis – development of the vascular supply within floral nectaries of Asteraceae
Davis, A 1 , Zhang, X 1 , Wist, TJ 2 , Sawhney, VK 1
1 Dept of Biology, University of Saskatchewan,
Saskatoon, Canada; 2 Dept of Biological Sciences,
University of Alberta, Edmonton, Canada
We are examining the development and role of vascular tissue within floral nectaries. The comprehensive surveys by Frei (1955) and Kartashova (1965) have demonstrated that floral nectaries of dicotyledonous angiosperms either are supplied by phloem alone, by both phloem and xylem, or lack any direct vasculature into the glandular tissue. We have found that in the floral nectaries of
Echinacea purpurea and Helianthus annuus , sieve tubes of phloem in the nectary interior connect continuously to sieve elements located within the subepidermal layer.
Moreover, evidence obtained by light and transmission electron microscopy suggests that phloem in the subepidermis of E. purpurea can originate from periclinal divisions of nectary epidermal cells, thus signifying a rare role of the epidermis in the formation of specialized vascular tissue. Companion cells are the only transfer cells found to exist within the nectaries of E. purpurea , and the presence of these wall ingrowths should assist determination of phloem-cell fate in the subepidermal layer. We seek to determine stimuli and patterns of actual formation of sieve elements and companion cells, plus a prospective role for invertase at the epidermis, to account for the hexose-richness of the floral nectar secreted through stomata on the nectary surface.
A role for auxin and PIN6 in the regulation of quantitative nectar secretion in the Brassicaceae
1
J 2
Bender, R 1 , Fekete, M 1
, Carter, C 1
, Bauer, B 1 , Lindgren, K 2 , Maki,
University of Minnesota Duluth, USA; 2 College of St
Scholastica, Duluth, USA
To date, a mechanistic understanding of nectar synthesis and secretion at the molecular level is lacking. In order to identify genes mediating nectary form and function, microarray analyses were used to identify nectaryenriched genes in Arabidopsis thaliana . One such gene,
PIN6, a polar auxin transporter, was confirmed to have extreme enrichment in nectaries by both RT-PCR and promoter:GUS fusion analyses. To identify a biological role for PIN6, three independent T-DNA mutant alleles were identified. Homozygous mutants were subsequently examined for altered expression level via quantitative reverse transcription polymerase chain reaction (qRT-
PCR). pin6-1, pin6-2 and pin6-3 mutants were identified as knock-up, knock-out, and knock-down mutants respectively. To link a role between PIN6 and nectary function, mutant flowers were analyzed for total nectar sugar, a measure of nectar production. Significantly,
PIN6 expression level was positively correlated to total nectar sugar, with pin6-1 flowers (knock-up mutant) displaying a 30% increase in nectar sugar over wild-type, and pin6-2 (knock-out) and pin6-3 (knock-down) having significant decreases in total nectar sugar. Interestingly, pin6-1 and pin6-3 flowers displayed no visible differences in morphology or nectary size from wild-type plants; however, pin6-2 flowers had petals that failed to fully expand, had reduced nectary size, and routinely lacked one or both short stamen. The flowers of pin6-2 plants complemented with a PIN6 expression construct
155

were fully restored to a wild-type phenotype, including total nectar sugar. In addition to the above, multiple T-
DNA mutants for other nectary-enriched genes, such as
MYB57 and gibberellin-2-oxidase 6 (GA-2-OX6), displayed reduced total nectar sugar. RT PCR analyses of these mutant lines also showed greatly reduced PIN6 expression level. Preliminary results indicated that
MYB57 directly binds to the PIN6 promoter and regulates its expression. Finally, to link auxin to quantitative nectar secretion, flowers treated with exogenous auxin displayed significant increases in total nectar sugar, whereas those treated with the auxin transport inhibitors TIBA and NPA produced little or no nectar. The findings described above are not unique to
Arabidopsis , as each has also been confirmed to occur in
Brassica rapa . A proposed mechanistic role for auxin and PIN6 in nectar production will be presented.
Protein profiles in floral nectar: beyond a simple reward
Nepi, M 1 , Guarnieri, M 1 , Pacini, E 1
1 University of Siena, Dept of Environmental Sciences,
Italy
Nectar is considered a common floral reward that can be utilized by a very large variety of animals. Sugars are the more abundant solutes present in nectar and they have been identified in a large number of species. Sucrose and its monomers fructose an glucose are almost ubiquitous as nectar sugars. Being simple molecules with a high energy content, easy to digest and absorb, they are considered the main alimentary reward of nectar. Other solutes, such as amino acids, vitamins, organic acids contribute to the alimentary value of nectar. Secondary compounds such as alkaloids and volatiles are involved in managing the insects behavior, being able to attract or repel them. Protein also have been detected in floral nectar since the early years of the last century but it is only more recently that nectar protein profiles have been better characterized. Apparently proteins are not involved in attracting or repelling animals but rather their role is related to two main functions: modifying the nectar sugar profile and protecting the nectar from microorganism invasion. Invertase is the enzyme responsible for the hydrolysis of sucrose into fructose and glucose and thus it is most involved in shaping the sugar profile of nectar.
It was the first enzyme known in nectar but its function was little characterized. Nectar is an excellent media in which fungi and bacteria can grow, especially when it is openly exposed to the environment. Thus the plant must defend this secretion and one way is the production of nectar defense proteins. Few studies demonstrated a large heterogeneity of nectar defense proteins in related and unrelated species. Both types of enzymes have recently been identified in the floral nectar of Cucurbita pepo : an invertase with a very low activity and four iso-forms of a
β
-D xylosidase for which a defense-related function can only be hypothesized for the moment. Although nectar of
C. pepo contains hydrogen peroxide up to 0.2 mM, a concentration that can be active against microrganisms, it was not ascertained if the nectar redox cycle is operating in this species. At present the study of nectar proteins by means of the modern techniques of sequencing and biochemical characterization is at its beginning but revealed already complex and heterogeneous functions
156 that could not be hypothesized before. The biochemical complexity of nectar demonstrate that this secretion, dynamically modulated by the nectary itself, is much more than a simple sugary reward for pollinators. This complexity opens new perspectives for future research.
Identification of new genes involved in
Arabidopsis cell wall expansion using activation tag screening
Somerville, C 1 , Anderson, C 1
1 University of California Berkeley, USA
Cell wall expansion, in which the wall must increase in surface area while retaining its ability to withstand turgor pressure, is essential for plant growth and development.
The expected early lethality of null mutations in genes required for cell wall expansion and the potential for redundancy to mask the phenotypes of mutations in gene family members limit conventional screening methods for mutants in this process. To circumvent these issues, we performed a screen to identify genes whose overexpression causes enhanced cell expansion. As an experimental system we chose etiolated Arabidopsis seedlings, in which hypocotyl elongation is driven primarily by cell expansion. From a population of
~30,000 lines transformed with an activation tag construct containing four tandem CaMV 35S enhancers, we identified putative single insertion lines with a heritable elongated hypocotyl phenotype. Identification of the insertion sites for these lines has allowed us to generate a list of candidate genes with potential functions in cell wall expansion. Characterization of these genes, many of which are of unknown function, should enhance our understanding of the molecular events of cell wall expansion and has the potential to inform efforts to more efficiently use plant cell walls for the production of biofuels and renewable materials.
Xyloglucan and glucomannan biosynthesis
Keegstra, K 1 , Cavalier, D 1
B 1 , Wang, Y 1
, Danhof, L
, Wilkerson, C 1
1 , Davis, J 1 , Reca,
1 Michigan State University, USA
Plant biomass is a major renewable resource that can be used in the production of biofuels and other chemicals. It consists mainly of plant cell wall polymers including cellulose, hemicellulose and lignin. In spite of their abundance in nature and their importance in the emerging biofuels industry, too little is known about the biosynthesis of these cell-wall components.
Hemicellulosic polysaccharides are known to be synthesized in the Golgi before their delivery to the cell surface. Identification and characterization of the enzymes require for the synthesis of hemicellulosic polysaccharides, such as xyloglucan or glucomannan, have been difficult because they are low abundance, integral membrane proteins. Consequently, we have pursued a functional genomics strategy, taking advantage of the genome sequence information that is available for

many plant species, including Arabidopsis . The proteins needed for the synthesis of xyloglucan or glucomannan from the various sugar nucleotides have been identified and partially characterized. However, many important questions regarding the molecular details of polysaccharide biosynthesis remain to be determined.
The lecture will review the work leading to the identification of the enzymes needed for xyloglucan and glucomannan biosynthesis and present current efforts to resolve some of the remaining unsolved problems. This research was supported with funding from the US Dept of Energy (DOE) Great Lakes Bioenergy Research
Center (DOE BER Office of Science DE-FC02-
07ER64494) and from the Chemical Sciences,
Geosciences and Biosciences Division, Office of Basic
Energy Sciences, Office of Science, U.S. Dept of Energy
(award no. DE-FG02-91ER20021).
Using forward genetic approach to study the monocot
1 cell wall biosynthesis
Zhang, B 1 , Xiong, G 1 , Li, R 1 , Liu, L 1 , Zhou, Y 1
Institute of Genetics And Developmental Biology,
Chinese Academy of Sciences,
China
Rice is a model organism for studying the mechanism of cell wall biosynthesis and remolding in Gramineae.
Mechanical strength and plant height are important agronomy traits of rice plants ( Oryza sativa L.) that affect crop lodging and grain yield. As the prominent physical property of cell walls, mechanical strength and plant height reflect upon the structure of different wall polymers and how they interact. Studies on the mechanisms that regulate these traits are therefore helpful for uncovering the functions of corresponding genes in cell wall biosynthesis and remodeling. Our group focuses on systematical isolation of mutants that have altered mechanical strength and/or plant height and functional characterization of their corresponding genes.
The identified genes cover several pathways of cell wall biosynthesis, including cellulose biosynthesis and deposition, membrane trafficking, matrix polysaccharides formation, and substrate supply. All of those have revealed many secrets of monocot cell wall biosynthesis and remodeling, which are of great benefit to harnessing the waste rice straws for biofuel production.
Insights into the synthesis, structure and function of the plant cell wall polysaccharide (1,3;1,4)-
β
-D-glucan
Doblin, M
C 1
1 , Burton, R
, Collins, H 2
2 , Jobling, S
, Kibble, N 2
3 , Wilson, S
, Fincher, G 2, 4
1 , Walsh,
, Bacic, A 1, 5
1 ARC Centre of Excellence in Plant Cell Walls, School of
2 Botany, University of Melbourne, Australia; ARC
Centre of Excellence in Plant Cell Walls, School of
Agriculture, Food and Wine, University of Adelaide,
Waite Campus, Glen Osmond, Australia; 3 CSIRO
4
Division of Plant Industry, Canberra, Australia;
Australian Centre for Plant Functional Genomics,
School of Agriculture, Food and Wine, University of
Adelaide, Waite Campus, Australia; 5 Australian Centre for Plant Functional Genomics, School of Botany,
University of Melbourne, Australia
Plant cell walls are of central importance to plants during growth and development, providing mechanical support, a reservoir of cell–cell communication and signaling molecules, and as a physical barrier to pathogen attack.
The non-cellulosic polysaccharide (1,3;1,4)-
β
-D-glucan, or
β
-glucan, is a plant cell wall polysaccharide found almost exclusively within the Poaceae and related families of the commelinoid clade of the Poales in higher plants. The walls of the Poaceae have been extensively studied because of the economic importance of grasses and cereals. In plants,
β
-glucans have been implicated in the control of cell expansion, acting as a structural element and as a carbon storage reserve. In the human diet, they are believed to lead to positive health benefits by lowering serum cholesterol and attenuating blood glucose levels. Soluble fibre is known to reduce the risk and occurrence of coronary heart disease, colorectal cancer, obesity and Type II diabetes. In contrast, because of the ability of
β
-glucans to form highly viscous solutions, they have a negative impact on the production and quality of beer by interfering with the filtration process and are also the cause of ‘sticky’ faeces in the feedstock industry. Hence, there is widespread interest in identifying the genes responsible for the synthesis, regulation and modification of
β
-glucan with the aim of being able to effectively produce cereals with high and low
β
-glucan levels for different end uses. We have identified two cellulose synthase-like gene families, CslF and CslH, as encoding the likely catalytic components of the (1,3;1,4)-
β
-D-glucan synthase enzyme (Burton et al.
2006, Doblin et al. 2009). We present our recent work in determining the other component/s and of the
β
-glucan synthesis machinery, the mechanism of enzyme action and the control of
β
-glucan fine structure as well as the regulation of its synthesis. This work was supported by funding from the Commonwealth Scientific and Research
Organization Flagship Collaborative Research Program, provided to the High Fibre Grains Cluster via the Food
Futures Flagship.
The biosynthesis of (1,3;1,4)-beta-D-glucan in the
1
Oryzeae tribe
Burton, R 1, 2
Cosse, M 1
, Fabrizio, J
, Little, A 2
1 , Collins, H
, Henderson, M 1
2 , Lahnstein, J 2
, Fincher, G 1,2
,
ARC Centre of Excellence In Plant Cell Walls,
University of Adelaide, Australia; 2 CSIRO Food Futures
National Research Flagship, University of Adelaide,
Australia
(1,3;1,4)-
β
-D-glucan is a non-cellulosic cell wall polysaccharide found in many cereal crops where it is an important source of dietary fibre for human nutrition providing substantial health benefits. It is found in high levels in certain cereals such as oats and barley but currently very little is known about the amount and distribution in rice, which is one of the world¡¦s most important cereal crops. Currently only data from grain at the very early stages of development, 3 to 5 days after pollination (DAP), has been published (Brown et al.
1997). Therefore, we have examined the amount and structure of (1,3;1,4)-
β
-D-glucan within economically important rice cultivars ( Oryza sativa ), and in a number of other Oryza species, in both developing grain samples
(6 to 28 DAP) and in various vegetative tissues using a sensitive high performance chromatography method. The
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physical distribution of this polysaccharide was also determined using microscopic analyses and the BG1 monoclonal antibody specific for (1,3;1,4)-
β
-D-glucan.
Transcript levels of (1,3;1,4)-
β
-D-glucan synthase genes, namely those in the CslF (Burton et al., 2006) and CslH
(Doblin et al., 2009) gene families, were analysed by real-time quantitative PCR using cDNA templates derived from vegetative tissues at various growth stages and from developing grain 6 to 28 DAP. Transcripts of
OsCslF6 predominated during mid grain development, which is consistent with the pattern previously reported for the direct barley and wheat orthologues (Burton et al.
2008, Nemeth et al. 2010). However, the detection of aberrant cDNAs in many rice tissues has led to an investigation into splicing patterns of particular CslF genes and the downstream effects this may have on protein function.
Genetics of cell wall polysaccharide synthesis in the grasses
Fincher, G 1
Russell, J 2
, Schwerdt, J 1
, Waugh, R 2
, Harvey, A 1 , Marshall, D 2 ,
1 ARC Centre of Excellence In Plant Cell Walls,
Australia; 2 Scottish Crops Research Institute,
Invergowrie, Dundee, Scotland, UK
The CsIF genes of barley and rice mediate the biosynthesis of (1,3;1,4)-
β
-glucans, which are noncellulosic cell wall polysaccharides found almost exclusively in the Poaceae. Six of the eight rice OsCsIF genes cluster on rice chromosome 7 within an interval of approximately 100 kilobases, while four of seven
HvCslF genes of barley, eight of the 11 sorghum SbCsIF genes and five of the seven BdCsIF genes from
Brachypodium map to a single locus in syntenic regions of these other species. Further, the CsIF gene clusters are components of ‘super clusters’ of genes that include groups of genes involved in plant responses to biotic stress. Within the clusters, individual CsIF genes exhibit
65–75% sequence identity, which suggests that the clusters have been conserved over a considerable time period. Thus, the CsIF gene clusters appear to be taxonomically conserved within the grasses but are nevertheless dynamic insofar as relatively recent expansions and contractions of cluster size can be detected. The conservation of the cluster might be attributable to selection pressure that allows the grasses to rapidly synthesise (1,3;1,4)-
β
-glucans during certain stages of development or in response to environmental stimuli.
MiR159 regulation of gene expression in
Arabidopsis
; specificity, mechanism and biological requirement
Li, J 1,2 , Lin, L 1 , Lin, Y 1
White, RG 2 , Gubler, F 2
, Alonso-Peral, MM
, Millar, AA 1
1 , Allen, RS 2 ,
1 Research School of Biology, Australian National
University, Australia; 2 Plant Industry, CSIRO, Australia
158
MicroRNAs (miRNAs) are a class of small RNAs that have been shown to control many critical aspects of plant biology. We are examining the miR159 class of miRNAs in Arabidopsis in order to understand general principles of the specificity and mechanism of miRNA-mediated gene silencing in plants. Firstly, bioinformatic and molecular analyses predicts miR159 to regulate over 20 genes in Arabidopsis , including eight MYB genes that contain a highly conserved miR159 binding site. Despite this, genetic analysis demonstrated that miR159 is functionally specific for only two target genes, MYB33 and MYB65, which have strong overlapping transcriptional domains with the two major miR159 members, miR159a and miR159b. Many of the other target genes have an overlapping transcriptional domain with the remaining family member, miR159c. However miR159c appears quiescent in nature, explaining the lack of functional impact of miR159 regulation of these predicted target genes. Secondly, with regards to mechanism, plant miRNAs have extensive complementarity and perfect matches around the cleavage site and so they are thought to act predominantly through mRNA cleavage, and to a lesser extent translational repression. However, using transgenic systems we show that that the steady-state levels of MYB33 do not correlate with silencing, despite the detection of miR159-guided cleavage products.
Moreover, artificial miR159 variants with mismatches around the cleavage site (both positions 10,11) can still fully silence MYB33. Together this suggests that miR159 repression of gene expression can work predominantly at the translational level. Finally we will speculate on the biological requirement of miR159 regulation, and the novel aspects that this type of posttranscriptional regulation can give, compared to regulation at a transcriptional level.
MicroRNAs in the shoot apical meristem of soybean
Wong, CE 1 , Zhao, Y-T 0 , Wang, X-J
Z-H 0
BJ 4,5
, Haerizadeh, F 1
, Bhalla, PL 1
, Mattick, JS 0
2 , Croft, L
, Singh, MB 1
0 , Wang,
, Carroll,
1 ARC Centre of Excellence for Integrative Legume
Research, Faculty of Land and Food Resources, The
University of Melbourne , Australia; 2 State Key
Laboratory of Plant Genomics, Institute of Genetics and
Developmental Biology, Chinese Academy of Sciences,
Beijing, China; 3 Graduate University of The Chinese
Academy of Sciences, Beijing, China; 4 ARC Centre of
5
Excellence for Integrative Legume Research , Australia;
School of Chemistry and Molecular Biosciences ,
Australia; 6 Institute for Molecular Bioscience, University of Queensland, Brisbane , Australia
Plant microRNAs (miRNAs) play crucial regulatory roles in various developmental processes. In this study, we characterize the miRNA profile of the shoot apical meristem (SAM) of an important legume crop, soybean, by integrating high-throughput sequencing data with miRNA microarray analysis. A total of 8,423 nonredundant sRNAs were obtained from two libraries derived from micro-dissected SAM or mature leaf tissue.
Sequence analysis allowed the identification of 32 conserved miRNA families as well as eight putative novel miRNAs. Subsequent miRNA profiling with microarrays verified the expression of a majority of these

conserved and novel miRNAs. It is noteworthy that several miRNAs* were expressed at a level similar to or higher than their corresponding mature miRNAs in SAM or mature leaf, suggesting a possible biological function for the star species. In situ hybridization analysis revealed a distinct spatial localization pattern for a conserved miRNA, miR166, and its star species suggesting they serve different roles in regulating leaf development. Further, localization studies showed that a novel soybean miRNA, miR4422a, was nuclearlocalized. This study also indicated novel expression pattern of miR390 in soybean. Our approach identified potential key regulators and provided vital spatial information towards understanding the regulatory circuits in the SAM of soybean during shoot development.
Defective embryo and meristems interacts with conserved Ras-like nuclear protein, and mutants display multiple epigenetic defects
Peters, J
L
V
0
3
1,2 , Gursanscky, N 0
, Reyes, M.E.C
0
, Kobe, B 1
, Ngo, Q 3
, Bowman, J.L
3,4
, Lonhienne, T 0
, Hawker, N 3
, Carroll, B.J
1,2
, Matthew,
, Sundaresan,
1 1School of Chemistry and Molecular Biosciences The
University of Queensland , Australia; 2 ARC Centre of
Excellence for Integrative Legume Research, The
University of Queensland ,
Biology, University of California at Davis, Davis, USA;
Clayton Campus , Australia
Australia; 3 Section of Plant
4 School of Biological Sciences, Monash University,
Coordinated cell proliferation and cell differentiation directs the shape and form of multicellular organisms.
The tomato defective embryo and meristems (dem) mutant fails to develop beyond the seedling stage of development. Somatically mutagenesis and modification of DEM expression, demonstrated that DEM directs both cell division and cell differentiation in tomato.
Arabidopsis has two DEM-like genes, DEM1 and
DEM2, and unlike dem in tomato, the single mutants, dem1 and dem2, remain viable. However, the dem1dem2 double mutant is a gametophytic lethal. DEM is specific to multicellular plants, and is not homologous to any protein of known biochemical function. However, a twohybrid screen identified Ras-like nuclear protein (RAN) as a potential interaction partner. E. coli -expressed proteins confirmed that DEM interacts with RAN in vitro. E. coli -expressed RAN proteins can also pull down
DEM from crude plant extracts. RAN is a guanine nucleotide binding protein that is highly conserved between plants and animals, and in animals has been shown to play key roles in: i) nucleo-cytoplasmic protein transport, ii) assembly of the spindle during mitosis, iii) formation of the nuclear envelop following mitosis, and iv) export of microRNAs (miRNA) from the nucleus.
Our recent work has demonstrated that Arabidopsis dem1 mutants have lower steady state levels of miRNAs. dem1 and dem2 mutants are also defective in transgene silencing. GFP fusions to the N- or C-terminus of DEM1 indicated that DEM is present in the cytoplasm, but restricted to the periphery of the nucleus. Our data are consistent with the hypothesis that the role of DEM in cell division and cell differentiation in multicellular plants is directed through its interaction with RAN, and its involvement in regulatory RNA pathways in the cell.
Introns protect transgenes from silencing in
Arabidopsis
Christie, M 1 , Carroll, BJ
1 School of Chemistry and Molecular Biosciences, The
University of Queensland , Australia
Transgene silencing is a major limitation on the expression of introduced genes in plants. The problem is exacerbated in applications that require expression of multiple transgenes in a stable and predictable fashion.
Here we show that introns can greatly reduce the frequency of transgene silencing in Arabidopsis .
Transformation of wild-type Arabidopsis with an intronless green fluorescence protein (GFP) transgene resulted in 85% of independent T1 transformants showing GFP silencing. Silencing was RDR6-dependent as 100% of independent rdr6 mutant transformants showed GFP expression. Initially, we tested four
Arabidopsis introns and found that three were able to significantly decrease the incidence of transgene silencing in primary transformants by up to five-fold.
Molecular characterization of these transgenic lines and others carrying additional modified introns showed that efficient splicing was required to provide protection against silencing. These results suggest that intron splicing protects a transcript from becoming a substrate for RDR6, and thereby protects it from gene silencing.
We have also shown that intron-mediated protection against silencing is dependent on ABH1, the Arabidopsis homologue of the human cap-binding protein 80
(CBP80). The mechanism of intron-mediated suppression of transgene silencing will be discussed.
A genome-wide survey of imprinted genes in rice seeds
Luo, M 1
Russell, S 3
, Taylor, J 1 , Zhang, H 2 , Spriggs, A 1
, Singh, M 4 , Koltunowe, A 5
, Wu, X 2 ,
1 CSIRO Plant Industry, Canberra, Australia; 2 Rice
Research Institute of Sichuan Agricultural University,
Chengdu, Sichuan, P.R. China; 3 Dept of Botany and
4
Microbiology, University of Oklahoma, Norman, USA;
Plant Molecular Biology and Biotechnology
Laboratory, The University of Melbourne, Parkville,
Australia; 5 CSIRO Plant Industry, South Australia,
Australia
Imprinting is a mechanism resulting in the biased expression of one allelic copy of a gene in a parent-oforigin specific manner. A few imprinted Arabidopsis genes are involved in regulating seed development. Only
17 imprinted genes have been confirmed in plants to date, therefore, we conducted a genome-wide survey of parent-of-origin-specific gene expression in rice seed, an important food crop. Transcriptomes of endosperm and embryos isolated from reciprocal crosses between two sequenced subspecies (Nipponbare and Chinese Indica
93-11) were profiled. Single Nucleotide Polymorphisms
(SNPs) enabled assignment of expressed sequences to the parental genome of origin. In the endosperm, 165 expression-biased genes were identified in addition to 35 expression-biased intergenic regions. 104 genes showed
>90% expression bias from a parental allele, of which 62 were maternally expressed and 42 paternally expressed.
159

Most of these genes are expressed lowly or show no evidence of imprinting in the embryo. Only one maternally expressed gene was found in the embryo and it was also maternally expressed in the endosperm. Rice imprinted genes show low conservation with known plant imprinted genes and they are not physically clustered or significantly associated with transposons and repeats relative to non-imprinted genes. Two cases of novel parent-of-origin alternative polyadenylation were identified in the endosperm. Four genes that were either biallelically expressed or partially imprinted early in endosperm development became uniparentally expressed, suggesting mechanisms operate in rice to establish imprinting after fertilization. We will discuss the putative mechanisms regulating imprinting in rice endosperm and their potential linkage to the small RNA pathway.
Do non-coding RNAs function in the
Arabidopsis vernalization response?
Helliwell, C 1 , Finnegan, J 1 , Robertson, M 1 , Dennis, L 1
1 CSIRO Plant Industry, Canberra , Australia
Expression of the Arabidopsis flowering repressor, FLC, is responsible for conferring vernalization responsive flowering in many late-flowering ecotypes. The repression of FLC by vernalization has the hallmarks of an epigenetically regulated response; FLC expression is repressed during cold and this repression is maintained once plants are returned to warm conditions. This repression of FLC requires the function of a PHD-PRC2
(Plant Homeodomain-Polycomb Repressive Complex 2) protein complex which adds the repressive H3K27me3 mark to FLC chromatin. A number of non-coding RNA transcripts have been identified at the FLC locus that show altered expression during the cold. These transcripts have been suggested to function in various aspects of the vernalization response. We will present the results of our investigations into the functions of these non-coding RNAs.
Mechanisms of mobile antiviral defence and
1 counterdefence in plants
Waterhouse, P
White, R 3
1 , Correa, R 2
, Anderssen, B 3
, Fusaro, A 1
, Carroll, B 4 , Kawchuk, L
University of Sydney, NSW, Australia;
Virologia, IMPPG, UFRJ, Brazil;
Industry, Canberra, ACT, Australia;
Queensland, Brisbane, QLD, Australia; 5
3
4
, Liang, D 3
6
2
,
Depto De
CSIRO Plant
University of
Agriculture and
Agri-Food Canada, Alberta, Canada
RNAi-like pathways are present in almost all eukaryotes and rely on small RNAs directing sequence specific RNA degradation, translational repression or epigenetic modifications within the cell. One of the RNAi pathways in plants mediates an adaptive mobile viral defense that has many parallels with the mammalian immune system.
Details of the mechanism by which the plant mobilizes and enhances its defense, and how members of one family of viruses, the Luteoviridae, have developed elegant, but different, counter-defence mechanisms, will be described.
Small RNAs in disease induction or symptom attenuation by viral satellite RNAs in plants
Smith, NA 1 , Eamens, AL 1 , Shen, W 1 , Zhou, C 1 , Wang,
1
M-B 1
CSIRO Plant Industry, Canberra, ACT, Australia
Plant viruses are often accompanied by small, 200–400nt parasitic RNAs termed satellite RNAs. These RNAs do not encode proteins and depend on the associated virus (helper virus) for their propagation and systemic spread. While the replication of many satellite RNAs attenuates the symptoms caused by their helper virus, some satellite RNAs, such as the cucumber mosaic virus
Y-satellite (Y-Sat), can itself induce disease symptoms in the host that are distinct to those of the helper virus. How these non-protein-coding RNA pathogens induce disease symptoms has been a long-standing question. We have previously reported that the yellowing symptoms induced by Y-Sat in tobacco was inhibited by the expression of a viral RNA silencing suppressor, which led us to suggest that pathogenic satellite RNAs might have sequence homology to a host gene, and that disease symptoms are due to satellite small interfering RNA (siRNA)-directed silencing of that host gene. No such host gene has been identified, leaving the satellite RNA-induced disease mechanism unsolved. Here we will present our recent finding that Y-Sat-induced yellowing symptoms in tobacco are caused by Y-Sat siRNA-directed silencing of a magnesium chelatase gene essential for chlorophyll biosynthesis. We also demonstrate that Y-Sat-induced disease symptoms can be prevented by the presence of a naturally evolved, or artificially introduced, silencingresistant sequence variant of the magnesium chelatase gene. These findings provide the first evidence of small
RNA-mediated viral disease symptom production, offer an explanation for the previously observed species specificity of satellite RNA-induced diseases, as well as providing a potential strategy to prevent such diseases.
We will also discuss our findings concerning the involvement of satellite RNA-derived siRNAs in the attenuation of helper virus-induced symptoms in plants.
Transgenic RNA silencing-mediated plant disease
1 control strategies
Mitter, N 1
The University of Queensland , Australia
RNA silencing is an innate defence response of plants to protect them against virus infections. Transgene-induced
RNA silencing is triggered by sequence-specific doublestranded RNA which can be derived from sense, antisense, hairpin RNA or artificial microRNA transgenes. Viral silencing suppressors have been shown to affect RNA silencing based transgenic virus resistance.
However, durable resistance has been achieved by targeting genes that encode viral RNA silencing suppressors, selection of homozygous plants or plants with multiple transgene copies, and pyramiding of target sequences from different viruses which may infect a particular crop. We have shown efficiency and stability of hpRNA-induced virus resistance in tobacco involving cucumber mosaic cucumovirus (CMV) and potato virus
Y potyvirus (PVY). We have shown that RNA silencing
160

mediated immunity against PVY in transgenic tobacco can be overcome by infection with CMV, making the plant susceptible to the virus it was engineered to resist.
A single hpRNA construct of converging sequences provided dual immunity to both CMV and PVY.
However, the location of the viral sequences in the construct affected effectiveness, with sequences flanking the intron acting as most efficient silencing inducers. In addition to viruses we are also investigating the use of plant-delivered hpRNAs to control fungal infections targeting Phytophthora root rot of avocado . The validation for the efficacy of dsRNA constructs targeting
P. cinnamomi has been obtained in Arabidopsis . We are also developing artificial microRNAs (amiRNAs) for introducing resistance to plant viruses. We have modified an Arabidopsis thaliana miR159 precursor to amiRNAs targeting viral mRNA sequences encoding the nucleocapsid protein (N) and the silencing suppressor
(NSs) genes of Tomato spotted wilt virus. Transient expression of amiRNAs in Nicotiana benthamiana has confirmed expression of virus-specific amiRNAs as well as the ability of the amiRNA constructs to confer resistance. The amiRNA-based resistance has been shown to be active even at lower temperatures, unlike siRNA-based resistance, and offers an effective approach to prevent breakdown of resistance in the field by expressing two to three amiRNAs targeting different essential regions of a virus. In addition, broad spectrum resistance to several viruses may also be achieved by coexpression of appropriately designed amiRNAs.
Identification of epigenetic components required for cell-to-cell movement of an RNA silencing signal in
Arabidopsis
Searle, IR 1 , Smith, L 2 , Baulcombe, DC 2
1 1Research School of Biology, ANU, Canberra ,
Australia; 2 Dept of Plant Sciences, University of
Cambridge, UK
RNA silencing is a sequence-specific RNA degradation process conserved in fungi, animals and plants that is associated with cell-to-cell movement of a mobile silencing signal. We developed a cell-to-cell movement of RNA silencing system in Arabidopsis using an inverted repeat transgene under the control of a phloemspecific promoter such that the spread of RNA silencing was manifested in regions around veins. We have previously identified epigenetic components, Polymerase
IV, RNA-Dependent-RNA-polmyerase 2, and CLSY (a
SNF2chromatin remodelling factor) that are required for mobile RNA silencing. In addition we also identified a putative histone H3 lysine 4 trimethyl demethylase,
JMJ14, that is required for mobile RNA silencing. In addition to an effect on mobile silencing the jmj14 mutants also had reduced CHH DNA methylation, increased abundance of endogenous transposon transcripts and they flowered earlier than wild type. We placed the activity of JMJ14 at a downstream point in
RNA silencing pathways because the subcellular locations of upstream components RNA-dependent RNA polymerase (RDR2) and Argonaute (AGO4) were not perturbed in jmj14 mutants. These results illustrate the potential for a link between RNA silencing and demethylation of histone H3 trimethylysine. We will also discuss our progress in identifying new mutants required for the spread of RNA silencing.
Genetic determinants involved in graft-transmissible gene silencing in
Arabidopsis
, Bowman, J L 2 , Carroll, B
1
Gursanscky, N 1 , Brosnan, C 1
J 1
School of Chemistry and Molecular Biosciences, and
ARC Centre of Excellence for Integrative Legume
Research, The University of Queensland, Brisbane,
2 Australia; Section of Plant Biology, University of
California at Davis, California, USA, and School of
Biological Sciences, Monash University, Clayton
Campus, Australia
We have been using Green Florescent Protein (GFP) as a reporter to study the mechanisms of graft-transmissible gene silencing in Arabidopsis . Recently, we developed a transgenic parent line for use in a forward genetic screen to identify additional genes involved in systemic gene silencing. This parent line carries a transgene expressing
GFP-specific dsRNA from a root tip-specific promoter along with a linked 35S:GFP transgene target. The GFP silencing phenotype of ungrafted plants of this line resembles the grafted plants described by Brosnan et al.
(2007), and rootstocks of this line transmit silencing to scions expressing GFP. Following mutagenesis, over 40 independent systemic silencing mutants were identified.
Several of these have been shown to be allelic to rdr6 and are deficient in both receiving the silencing signal in newly formed shoot tissue and in transmission of the mobile silencing signal from grafted rootstocks. RDR6 is part of the trans-acting siRNA pathway, and additional components of this pathway are being tested for involvement in transmission of silencing signals from rootstocks. We are using map-based gene cloning to identify another mutant gene that does not appear to be linked to any genes known to be required for grafttransmissible gene silencing.
MIDD1: a novel membrane-associated MAP regulating secondary wall patterns
1
Fukuda, H 1 , Oda, Y 1
University of Tokyo, Japan
To analyze cellular events visually underlying xylem formation, we have established an Arabidopsis culture system in which a master transcription factor, VND6, is induced by a steroid hormone. In these systems, approx.
80% of cells differentiate to metaxylem vessel-like cells, within 3 days. Using this culture, we performed transcriptome analysis and found a number of genes whose function is not understood yet. The VND6inducive culture also allowed us to follow cellular events as live images in differentiating xylem cells under a microscope. With advantages of our newly established in vitro xylem cell differentiation system, we analyzed the function of products of unknown genes and discovered a novel microtubule end-tracking protein, designated
161

MIDD1 (Microtubule Depletion Domain 1). This protein was anchored to distinct plasma membrane domains and promoted local microtubule disassembly, resulting in pits on walls of metaxylem vessel cells. The introduction of
RNAi for MIDD1 resulted in failure of local microtubule depletion and the formation of secondary walls without pits. Conversely, the overexpression of MIDD1 reduced microtubule density. MIDD1 has two coiled-coil domains; the first domain associates with microtubules and the second domain is required for the anchorage of
MIDD1 to distinct plasma membrane domains.
Combination of the two coils caused end-tracking during shrinkage and promoted microtubule disassembly. Our results indicate that plants use a distinctive protein that integrates spatial information in the plasma membrane with cortical microtubule dynamics, for determining xylem cell wall pattern.
Turning over tracks: dynamics of actin filaments and cables in the cortical array of
Arabidopsis
epidermal cells
Staiger, C 1 , Henty, J 1 , Sheahan, M 2
1
McCurdy, D 2 , Blanchoin, L 3
, Khurana, P 1 ,
Purdue University, USA;
Australia; 3
2 Newcastle University,
CEA/CNRS/UJF, Grenoble, France
Eukaryotic cells harness the power of actin dynamics to create cytoskeletal arrays that stimulate protrusions and drive intracellular organelle movements. In plants, the actin cytoskeleton is generally understood to participate in cell elongation and responses to biotic and abiotic stimuli; however, a detailed description and molecular mechanism(s) underpinning filament nucleation, growth and turnover are lacking. We have used variable-angle epifluorescence microscopy (VAEM) to examine the organization and dynamics of the cortical cytoskeleton in growing and non-growing epidermal cells from
Arabidopsis hypocotyls. Actin in the cortical array exists as individual actin filaments that are short-lived, as well as longer-lived actin filament bundles. Collectively, the single actin filaments are randomly oriented and surprisingly dynamic. Single actin filaments grow at rates of 1.7 µm/s. Instead of depolymerization at their ends, actin filaments are disassembled by prominent severing activity. Incessant remodeling of the cortical actin array also features filament buckling and straightening events. We consider several mechanisms for the control of actin dynamics, including rapid polymerization from a large pool of profilin–actin, specific severing and capping activities, and myosindriven filament-filament interactions. Aspects of this model have been tested with pharmacological agents and with reverse-genetics. Specifically, we have demonstrated a role for ADF4 in severing actin filaments in vivo. And, we find that myosin XI contributes to actin dynamics. Our observations, the first to describe single actin filament behavior in plant cells, indicate a mechanism inconsistent with treadmilling, instead resembling the stochastic dynamics of a recently described biomimetic system for actin assembly in vitro.
Myosin XI drives endoplasmic reticulum motility and organizes F-actin orientations
Ueda, H 1 , Yokota, E 2 , Kutsuna, N
4 , Hara-Nishimura, I 1
3 , Shimada, T 1
3
1
VV
Kyoto University, Japan; 2
, Dolja,
University of Hyogo, Japan;
The University of Tokyo, Japan; 4 Oregon State
University, USA
Plants exhibit an ultimate case of the intracellular motility involving rapid organelle trafficking and continuous streaming of the endoplasmic reticulum (ER).
Although it was long assumed that the ER dynamics is actomyosin-driven, the responsible myosins were not identified, and the ER streaming was not characterized quantitatively. Here we developed software to generate a detailed velocity distribution map for the GFP-labeled
ER. This map revealed that the ER in the most peripheral plane was relatively static, whereas the ER in the inner plane was rapidly streaming with the velocities of up to
~3.5
µ m/sec. Similar patterns were observed when the cytosolic GFP was used to evaluate the cytoplasmic streaming. Using gene knockouts, we demonstrate that the ER dynamics is driven primarily by the ERassociated myosin XI-K, a member of a plant-specific myosin class XI. Furthermore, we show that the myosin
XI deficiency affects organization of the ER network and orientation of the actin filament bundles. Collectively, our findings suggest a model whereby dynamic threeway interactions between ER, F-actin, and myosins determine the architecture and movement patterns of the
ER strands, and cause cytosol hauling traditionally defined as cytoplasmic streaming. (Ueda et al. 2010).
Auxin regulation of cytoskeletal organization, endomembrane trafficking and cell morphogenesis in
Arabidopsis
Yang, Z 1 , Nagawa, S 1 , Xu, T 1 , Lin, D 1
1 University of California, Riverside, USA
Auxin is a universal morphogenetic signal that regulates the formation of various developmental and morphogenetic patterns in plants, but the molecular and cellular mechanisms for the auxin action remain poorly understood. We investigate these mechanisms using
Arabidopsis leaf epidermal pavement cells as a model system, which form the jigsaw puzzle cell pattern with interdigitated lobes and indentation. We have shown that pavement cell interdigitation is controlled by localized
ROP GTPase signaling, which impinges on the organization of cortical actin microfilaments and microtubules to generate interdigitating lobes and indentations (1, 2). Our studies indicate that auxin is a signal that activates pavement cell polarization to form lobes and coordinates lobe formation with indentation formation by activating ROP GTPase signaling through an Auxin-Binding Protein 1 (ABP1)-dependent and cell surface-based auxin perception system (3). This is a new cytoplasmic auxin signaling mechanism distinct from the well-established TIR1-dependent nuclear auxin signaling system that regulates gene transcription. This new auxin signaling mechanism regulates the interdigitated cell pattern through its modulation of PIN1 localization to lobe tips (3). Our recent results suggest that ABP1/ROP
162

signaling regulates PIN1 localization by affecting PIN1 endocytosis ad its recycling. These findings may provide new insights into the molecular and cellular mechanisms for auxin action as well as the first glimpse at how cellular signaling links the cytoskeleton with endomembrane trafficking in plant cells. References: (1)
Fu, Y et al. 2005. (2) Fu, Y et al. 2009. (3) Xu T et al.
2010.
An
Arabidopsis
formin tracks microtubule dynamics and is involved in cell division
Ren, H 1 , Li, Y 1 , Shen, Y 1 , Cai, C 1
1 Beijing Normal University, China
Formins have long been known to regulate microfilaments, but have also recently been shown to associate with microtubules. We have previously studied a type II formin from Arabidopsis thaliana –AtFH14 – and found it regulated both microtubule and microfilament arrays. AtFH14-GFP expressed in BY-2 cells was shown to decorate preprophase band (PPB), spindle, and phragmoplast, and to induce co-alignment of microtubules with microfilaments. Pharmacological experiments with cytoskeleton disrupting drugs showed that AtFH14 bound preferentially to microtubules.
Knockdown of AtFH14 in mitotic cells altered interactions between microtubules and microfilaments, resulting in the formation of an abnormal mitotic apparatus. To find out which domain is important for the localization of AtFH14, we expressed its N-terminal Pten domain and FH1FH2 domain in BY-2 cell and found both of them could target to PPB, spindle and phragmoplast, indicating that the location and function of
AtFH14 was determined by both Pten and FH1FH2 domains. To explore the function and localization of
AFH14 in non-dividing cells, we expressed FH1FH2-
RFP in onion epidermal cells, and found a fluorescence labeled filamentous network. The results of double labeling with different cytoskeleton reporter proteins indicated that FH1FH2-RFP co-localized with cortical microtubules. Treatment of cells expressing FH1FH2-
RFP with cytoskeleton disrupting drugs confirmed that
FH1FH2-RFP bound to microtubules. Moreover, the binding of FH1FH2-RFP to microtubules were revealed to be dynamic by fluorescence recovery after photobleaching experiment. Time-lapse confocal microscopy showed that FH1FH2-RFP could display a dynamics similar to the microtubule dynamic instability.
These results suggest that AtFH14 is a unique plant formin that tracks microtubule dynamics and is involved in cell division.
ACTIN7 is specifically required for chloroplast repositioning in dedifferentiating plant cells
Sheahan, M 1
McCurdy, D 1
, Kandasamy, M 2 , Meagher, R 2 , Rose, R 1
1 Newcastle University, Australia;
USA
2
,
University of Georgia,
The actin cytoskeleton facilitates numerous cellular processes required for the correct functioning and development of multicellular eukaryotes. Unlike yeast, where actin is encoded by a single gene, in multicellular eukaryotes actins are encoded by multi-gene families. In
Arabidopsis , eight expressed actin isoforms exist. Based on their phylogenetic relationship and expression pattern,
Arabidopsis actins are classified as either vegetative or reproductive, with each class being expressed predominantly in vegetative and reproductive tissues, respectively. The vegetative class of actins comprises
ACT2, ACT7 and ACT8. Interestingly, the sequence divergence between individual actin isoforms in plants is greater than the divergence between actin isoforms in animals, suggesting there is a potential for plant actin isoforms to perform discrete functions within the cell.
Alternately, such divergence may simply reflect developmental rather than intracellular specialisation. In this regard, we have been investigating chloroplast partitioning before the first cell division of Arabidopsis mesophyll protoplasts. In protoplasts of a number of plant species we have examined, chloroplasts cluster around the nucleus in an actin-dependent manner before the cell divides. This repositioning acts as a mechanism to ensure unbiased chloroplast inheritance. We have shown that chloroplast repositioning occurring before the re-initiation of cell division in cultured plant cells is defective in Arabidopsis plants carrying mutations in
ACT7 but not ACT2 or ACT8. Here, we investigated whether this apparent differential requirement for actin isoforms reflects differential expression or a specific functional requirement for ACT7 in chloroplast repositioning. Quantitative real-time PCR confirmed that
ACT7 was upregulated, while the two other vegetative actins, ACT2 and ACT8 were down-regulated slightly during protoplast culture. Among the reproductive actins, expression of ACT4 and ACT12 was not detected;
ACT11 expression remained relatively stable, whereas expression of ACT1 and ACT3 was upregulated.
Quantitative immunoblotting using cultured protoplasts derived from wild-type and act2-1 or act7-1 mutant backgrounds revealed an increase in total actin as protoplasts culture proceeded. Expectedly, there were reduced levels of total actin in the act7-1 mutant, yet total actin did increase during culture, indicating that other actin isoforms are present in these cells. Currently, we are examining chloroplast repositioning in act7-4 mutants expressing either ACT2 or ACT8 under control of the
ACT7 promoter to determine if the requirement for
ACT7 represents a developmental specialisation for this isoform or whether is simply represents the most abundantly expressed actin. Further, we report on the structural features of the actin cytoskeleton in wild-type, act2-1, act7-4 and act8-1 mutant backgrounds.
Novel and ancient aspects of flowering plant pollen
1 tubes and their pathways
Williams, J 1
Dept of Ecology and Evolutionary Biology, University of Tennessee, USA
Early angiosperm history was strongly marked by a shift from a long to an exceptionally brief fertilization process.
Ovules and their associated female gametophytes became
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greatly reduced in size, causing precocious production of an egg, and consequently a much abbreviated pollination to fertilization period (progamic phase). The shift to small ovules was accompanied by the origin of an enclosing carpel as well as the flower, which was itself small and ephemeral. Early angiosperm pollen tube evolution occurred within the context of many developmental modifications that affected the size and longevity of pollen tube pathways. A central innovation underlying the integration of interdependent floral and ovular tissues was the evolution of faster pollen tube growth rates. I studied pollen tube development and growth in a number of ancient lineages of angiosperms, including Amborella , Nymphaeales and Austrobaileyales.
I used ancestral reconstruction methods on these and other published studies to infer ancestral features of early angiosperm progamic phase biology. These reconstructions have revealed a surprising connection between the evolution of the microscopic fertilization process and ecological traits such as the duration of reproductive cycles and floral form and size. A survey of
275 angiosperm species shows that in vivo pollen tube growth rates range from 0.044 – 24 mm/h (median =
0.840 mm/h). Among ancient angiosperms, woody perennials have among the slowest pollen tube growth rates known, whereas a number of ancient aquatic lineages are characterized as having growth rates near or higher than the angiosperm median. Ancient angiosperm pollen tubes develop in a great diversity of environments: they germinate in water or on dry or wet stigmas, and tubes grow in water, free secretions, secretions between appressed tissues, zones of secondary fusion, and within middle lamellae between cells. Pollen tube structure is less variable. Ancient angiosperm pollen tubes are narrower in diameter and have thinner walls than those of other seed plants. Their thin walls are composed primarily of callose, whereas those of gymnosperms and most other plant cells are generally pecto-cellulosic and have a different pattern of callose expression, if present.
Callose is known to be deposited more rapidly than cellulose and has mechanical properties that support long-distance growth. It is synthesized in pollen tube walls and callose plugs via the same gene copy (CalS-5) in Arabidopsis as in ancient angiosperms such as
Amborella . CalS-5 is itself an ancient gene and is expressed in the intine of pollen of Ginkgo and Zamia . It was likely co-opted from a role within the pollen grain to its novel structural function in the pollen tube wall. Early increases in pollen tube growth rates evolved by streamlining of ancestral features (size changes) and by the origin of novelties (the permanent callose wall).
Subsequent changes must also have involved increases in rates of synthesis of callose and other wall materials.
Ultimately, rapid pollen tube growth rates enabled a dissociation of the pollen reception apparatus (the stigma) from the egg-bearing structure (the ovule), making possible a tremendous diversification in sizes and lifespans of floral and fruit organs.
Ions, energy, actin and pectin in the control of pollen tube growth
Hepler, P 1 , Rounds, C 1
1
Winship, L 2
, Kunkel, J 1 , Shipley, A 1 ,
2
University of Massachusetts, Amherst, MA, USA;
Hampshire College, Amherst, MA, USA
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Multiple coordinated processes are essential for the deposition of new membrane and wall material at the apex of the pollen tube, allowing rapid growth of the cell through the style and delivery of the two sperm cells to the embryo sac. Here we focus on four processes in lily pollen tubes; ion flux (calcium and protons), energy transduction, the structure and role of actin filaments, and the delivery, incorporation, and covalent modification of pectins. Our primary method is time lapse imaging with high resolution microscope optics coupled with specific endogenous or introduced fluorescent probes. We further exploit the oscillatory character of pollen tube growth, which is accompanied by a corresponding oscillation in the structure and/or activity of these underlying factors.
Calcium and protons show specific profiles in the tube apex. Calcium forms a steep tip-focused gradient, in which the concentration is high at the apical plasma membrane and declines to basal levels of 150 nM, within
20
µ m. During oscillatory growth the concentration changes from 750 nM to above 3,000 nM with a period of 15–50 sec. Phase analysis reveals, however, that the increase in calcium follows the increase in growth rate, suggesting that its changes do not stimulate growth.
Proton imaging yields a more complex pattern wherein the extreme apex is slightly acidic, and the base of the clear zone possesses an alkaline band. Analysis of the oscillatory activity indicates that the increase in pH of the alkaline band, presumably driven by the plasma membrane localized proton ATPase, anticipates the increase in growth rate and may control other transport processes that are essential for growth. Cellular energy status, monitored as the endogenous fluorescence of reduced mitochondrial NAD(P)H, reveals that maximal
NAD(P) + precedes the increase in growth rate. These observations fueled the idea that a corresponding increase in ATP might be pivotal in driving oscillatory growth. However, inhibition of the mitochondrial electron transport chain only momentarily blocks growth; cell elongation soon resumes and exhibits oscillations, which are not accompanied by oscillations in NAD(P)H.
Further studies reveal that the challenged pollen tubes produce ethanol, indicating that their metabolism shifts from oxidative phosphorylation to aerobic fermentation.
The presence of a prominent actin fringe in the pollen tube apex has led to speculation that these microfilaments transport and direct vesicles to fusion sites on the plasma membrane. However, inhibition and reinitiation studies reveal that polarized growth precedes the appearance of the actin fringe. Cell wall deposition emerges as a possible polarizing factor, wherein the newly thickened wall biases the direction of future growth. In addition, the deposition of wall material at the tip anticipates the increase in growth rate. Statistical analysis indicates that the magnitude and extent of wall deposition predicts to a high degree the subsequent growth profile. We suggest that the intercalation of newly secreted pectins into the existing wall relaxes the wall structure and allows for turgor dependent expansion of the cell. The location and rate of pectin exocytosis and intercalation may directly control growth rate and direction.

Space and time coordination of cellular growth processes in pollen tubes
Feijo, J 1
1 Dep. Biologia Vegetal, Fac.Ciencias, Universidade
Lisboa, Portugal
Pollen tubes are favourable models for fundamental understanding of cellular growth and morphogenesis in apically growing cells. Transcriptomics reveals the expression of about 7.000 genes, but theoretical modeling suggests that the cooperation of all of these into the processes of wall surface and cytoplasmic volume production, is a minimal condition to explain most of the morphogenic events that characterize these cells. Spatial and temporal integration of extended biochemical and biophysical processes is mandatory, and in the past we have proposed and demonstrated that ion dynamics can be a common regulator of fundamental growth processes. In order to test this hypothesis we are developing a number of genetic, imaging and electrophysiological approaches to define the set of membrane transporters that could underlie the transduction necessary for spatial and temporal coordination. We have uncovered original data in terms of proton pumping, and will describe new mechanisms for calcium and chloride (anion) transport. We will provide data suggestive that the feed-back mechanisms by which these ions could affect fundamental cell biology mechanisms is centered on the membrane recycling mechanisms. In fact advanced imaging methods and Monte-Carlo simulations suggest that sorting of exocytic vesicles might be achieved by direct electric interaction with ion fluxes. Finally we developed stringent 3-D theoretical modeling of ion fluxes and cytosolic diffusion based on the current knowledge of the system. These models are instrumental to define the minimal needs for channels to explain all the available evidence. Hopefully they will allow us to expand these conclusions to a broader understanding of the fundamental basis that govern cellular growth and morphogenesis by directed exocytosis, and will give us new insights on sexual plant reproduction, namely on the evolutionary advantage that pollen tubes rapid growth seemed to have given to angiosperm radiation and overwhelming success, Darwin's 'abominable mystery'.
S-RNase-based self-incompatibility: a complex nonself recognition system between pollen and pistil
Kao, T-H 1,2
Fields, AM 2
, Wang, N
, Hua, Z 3
1 , Sun, P 1 , Li, S 1 , Natale, CA 2 ,
1 Intercollege Graduate Degree Program In Plant
Biology, Penn State University, USA; 2 Dept of
Biochemistry and Molecular Biology, Penn State
University, USA; 3 Dept of Genetics, University of
Wisconsin, Madison, USA
Self-incompatibility (SI) possessed by Petunia inflata is controlled by a highly polymorphic locus, named the Slocus, which contains the genes that encode pollen and pistil specificity determinants in SI interactions. Our lab showed that the S-RNase gene controls pistil specificity
(Lee et al. 1994) and that the RNase activity of S-RNase is essential for its function in SI, suggesting that degradation of pollen tube RNAs is responsible for growth inhibition of self-pollen tubes (Huang et al.
1994). Our lab identified the PiSLF ( P. inflata S-locus Fbox) gene from sequencing a 328-kb region of the S2locus containing S-RNase (Wang et al. 2004), and established its SI function in pollen by showing that expression of PiSLF2, the S2-allele of PiSLF, in pollen of S1S2 and S2S3 transgenic plants caused breakdown of
SI in S1 and S3 pollen (heteroallelic pollen), but not in
S2 pollen (homoallelic pollen), a finding as predicted by competitive interaction (Sijacic et al. 2004). We proposed a model predicted on the preferential non-self interactions between PiSLF and S-RNase resulting in specific degradation of non-self S-RNases inside a pollen tube (Hua and Kao 2006). We further tested the function of two additional PiSLF alleles, PiSLF1 and PiSLF3, in transgenic plants of S2S3 genotype. The results, along with those obtained by the lab of Professor Seiji
Takayama (Nara Institute of Science and Technology,
Japan), led to the discovery that the pollen specificity determinant is encoded by multiple types of SLF genes, with each type of SLF responsible for detoxifying a different subset of non-self S-RNases (Kubo et al. 2010).
For example, for S2-haplotype, Type-1 SLF (old name
PiSLF) interacts with S1-RNase and S3-RNase, but not with S6-RNase, which interacts with Type-8 SLF (old name PiSLFLb), and no type of SLF interacts with self-
S-RNase, S2-RNase. We have set out to determine the biochemical basis that allows a particular type of SLF to interact with certain non-self S-RNases, but not with other non-self S-RNases or self S-RNase. The approach is to express chimeric SLFs between different types of
SLFs, and between different alleles of the same type of
SLF, in appropriate S-genotypes of transgenic plants, and to analyze the SI behavior of the transgenic plants. We are also testing the effect of suppressing the expression of a particular type of SLF on the SI behavior of the transgenic plants.
FERONIA receptor kinase controls RAC/ROP
GTPase-mediated and ROS-regulated root hair growth and pollen tube–ovule interaction
Cheung, AY 1 , Duan, Q 1 , Kita, D 1 , Johnson, E 1
1
M 1
University of Massachusetts, USA
, Wu, H-
RAC/ROP (plant RHO) GTPases are activated by guanine nucleotide exchange factors (ROPGEFs) and regulate diverse cellular, growth and developmental processes. We showed in 2002 and 2005 (1, 2) that auxin rapidly activates Rac/Rops which in turn mediate auxinresponsive gene expression. Using ROPGEF1 from
Arabidopsis as bait, we identified the FERONIA (FER) receptor kinase and a few other related kinases as
ROPGEF1 interacting proteins. Using feronia (fer) mutants, we showed that (3) FERONIA regulates
RAC/ROP-mediated, reactive oxygen species (ROS)dependent and auxin-responsive root hair development, defining a cell surface receptor to a well established
RAC/ROP signaling pathway (4). FER was previously identified as an important regulator for female gametophytic function (see 5). Loss of function fer mutants fail to support pollen tube rupture upon entrance into the female gametophyte, resulting female sterility and a dramatic pollen tube overgrowth phenotype.
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Furthermore, fer mutants fail to deter later arriving pollen tubes, resulting in supernumerary pollen tube penetration of the female gametophyte. We will discuss recent results that show FER functions in the female gametophyte via controlling ROS production in the synergid cells and that
ROS are crucial for pollen tube rupture. FER also regulates pectin deposition at the filiform apparatus gating the female gametophyte, providing a possible gating mechanism to regulate pollen tube penetration.
References (1) Tao et al. 2002. (2) Tao et al. 2005. (3)
Duan et al. 2010. (4) Carol et al. 2005. (5) Escobar-
Restrepo et al. 2007. This work was supported by grants from NSF (IOB0544222) and USDA (CSREES 2004-
35304-14837).
Molecular controls of karyogamy in
Arabidopsis
Berger, F 1 , Kawashima, T 1 , Jet Aw, S 1
1 Temasek LifeScience Laboratory, NUS, Singapore
Fertilization in flowering plants involves two sperm cells and two female gametes, the egg cell and the central cell, progenitors of the embryo and the endosperm, respectively. The mechanisms triggering zygotic development are unknown and whether both parental genomes are required for zygotic development is unclear.
Using the mutant in the CYCLIN DEPENDENT
KINASE A1 (CDKA;1) that impedes cell cycle progression and other mutants we present evidence for the control of karyogamy and activation of the zygote.
We monitor in vivo the fusion of the male and female nuclei using fluorescent markers to study the expression of the genome of each parent after fertilization. Our results support that both parental genomes are expressed in the zygote and that the paternal genome plays an essential role during early seed development. We also study the inheritance of parental chromatin in connection with transcriptional reactivation and report the essential role played by H3.3 variants in this step.
Identification of primary regulated genes – a genomewide approach defining specific nuclear target genes responsive to retrograde photosynthetic redox signals in
Arabidopsis
Pfannschmidt, T 1 , Dietzel, L 1
1 Plant Physiology, Friedrich-Schiller-University Jena,
Germany
In plant population such as forests or crop fields strong and persistent light gradients occur which require a structural and functional acclimation of the photosynthetic apparatus in order to maintain photosynthetic efficiency. A specific long-term response
(LTR) to light quality gradients is the adjustment of photosystem stoichiometry. This requires a controlled change in the expression of both nuclear and chloroplast genes encoding the constituents of the photosynthetic apparatus. Transcript profiles from Arabidopsis show that several nuclear genes are coupled to chloroplast expression events and that redox signals from the photosynthetic electron transport chain represent a new and important class of retrograde signals. Furthermore, the data suggest that photosynthetic acclimation is connected with fundamental biosynthesis pathways of the cell highlighting a possible conjunction between energy fixation efficiency and energy usage by metabolism.
Metabolite profiling confirmed that photosynthesis controls also the metabolic states of the plant.
Acclimation of photosynthesis into so-called state 1 or state 2 is thus accompanied by a corresponding adjustment of the metabolic states. Photosynthetic redox signals are proposed to integrate two adjustment loops which control and coordinate photosynthesis gene expression and metabolite fluxes. We aimed to understand these complex regulation mechanisms by identifying those nuclear genes responding directly to plastid redox signals within 30–60 minutes. We set up an approach in which we combined a physiological light system that induced timely defined redox signals with a comparative analysis of the full-genome expression profiles of Arabidopsis wild-type and stn7 mutant plants in response to the signal. The stn7 mutant is deficient in photosynthetic acclimation due to the lack of the key sensor kinase STN7. By identifying the genes responsive in WT but not in the mutant we were able to define for the first time the precise nuclear complement of redox controlled genes without using chemical inhibitors.
These primary regulated genes provide tools for constructing novel screens that will help to identify protein factors potentially involved in the transmission of the redox signal from the thylakoid membrane into the nucleus.
Regulation of photosynthesis-related nuclear gene expression by plastid retrograde signals
1
Gray, J
Tran, L 1
1 , Cottage, A
, Newell, C 1
1 , Mott, E 1
, Aspinall, S 1
, Lee, Y-C 1 , Jouhet, J 1
, Kavanagh, T 2
University of Cambridge, UK; 2
,
Trinity College Dublin,
Ireland
The expression of nuclear genes encoding components of the photosynthetic apparatus in higher plants is dependent on the developmental and functional state of the plastids. Mutations or inhibitor treatments affecting carotenoid synthesis or plastid gene expression result in the down-regulation of photosynthesis-related nuclear genes. Microarray analysis of transcripts in Arabidopsis seedlings treated with norflurazon, a carotenoidbiosynthesis inhibitor, or lincomycin, a plastid protein synthesis inhibitor, identified ACC2, encoding a plastidlocated homomeric acetyl-CoA carboxylase, as a gene that is highly up-regulated by norflurazon or lincomycin treatment. Treatment of barley and Arabidopsis seedlings with inhibitors of plastid acetyl-CoA carboxylase or fatty acid synthesis resulted in decreased expression of photosynthesis-related nuclear genes suggesting they are responsive to the state of plastid lipid synthesis. The
Arabidopsis genomes uncoupled1 (gun1) mutants are able to express photosynthesis-related nuclear genes in the presence of norflurazon or lincomycin, and are hypersensitive to sucrose and abscisic acid (ABA) during early seedling development (Cottage et al., 2010). GUN1 is a plastid nucleoid-associated pentatricopeptide repeat
(PPR) protein (Koussevitzky et al., 2007; Cottage et al.,
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2008) that has been proposed to integrate signals generated by norflurazon, lincomycin and high-light treatments (Koussevitzky et al., 2007). We are currently exploring the hypothesis that each of these treatments, and others resulting in down-regulation of photosynthesis-related nuclear genes, generates the same molecular signal that is sensed by GUN1.
Searching for functions of plastid-localised pentatricopeptide repeat proteins that contain a small
MutS-related domain
Howell, KA 1 , Liu, S 1 , Small, ID 1
1 ARC Centre of Excellence In Plant Energy Biology,
Crawley, Australia
The phenomenon of retrograde regulation whereby signals emanating from the plastids or the mitochondria, reflecting their functional and developmental state, exert a regulatory effect on nuclear genes is crucial to plant growth and survival. While this process has been extensively investigated, the exact molecular mechanisms underlying this form of regulation are not well understood (1). With respect to plastid-to-nucleus retrograde signalling, it has been proposed that this form of intracellular communication involves: plastid gene expression, redox signals, the tetrapyrrole pathway and/or reactive oxygen species (2). Screening for genome uncoupled (gun) mutants has identified some components specific to these pathways and one gun phenotype has been attributed to mutations in a gene encoding a member of the large pentatricopeptide repeat
(PPR) protein family (3, 4). In addition to PPR domains this protein, GUN1, also contains a small MutS-related
(SMR) domain, which in bacteria has been implicated to have DNA-binding and endonuclease activity (5,6).
Thus, it has been suggested that GUN1 could potentially bind both RNA and DNA. However, while GUN1 has been shown to play a central role in plastid-to-nuclear signalling, its exact function remains elusive.
Bioinformatic analysis of the Arabidopsis genome reveals that there are other PPR proteins with this domain structure and 5 of these (including GUN1) are predicted or have been found to be plastid-localised. We have initiated functional characterisation of other members of this PPR subfamily to determine what role they play in plastid function and plastid-to-nuclear signalling. Our investigations have involved the characterisation of mutant plants including analysis of gross phenotype, nuclear and organellar DNA levels, plastid transcripts and phenotypic and genetic responses to treatments affecting plastid function. Work has also been initiated to identify nucleic acid targets and further investigate the roles of the specific protein domains by testing complementation of mutants by truncated and fusion proteins derived from different combinations of the PPR and SMR domains. Using these approaches we hope to shed light on the function of the plastid-localised PPR-
SMR proteins and their role in retrograde signalling pathways and plastid function. (1) Kleine et al.
2009. (2) Pogson et al. 2008. (3) Koussevitzky et al.
2007. (4) Cottage et al. 2008. (5) Moreira and Philippe
1999. (6) Fukui et al. 2007.
Identification of the components mediating mitochondrial retrograde regulation in
Arabidopsis thaliana
1
Ivanova, A 1 , Law, S 1
Whelan, J 1
, Ng, S 1 , Van Aken, O 1 , Giraud, E
The University of Western Australia, Australia
1 ,
Retrograde regulation describes adaptive regulatory pathways from organelles, mitochondria and plastids, to the nucleus. This type of regulation is present in both single and multi-cellular eukaryotes. It allows organelles to signal specific needs or challenges to the nucleus, and thus is important for sensory responses to both developmental and environmental cues, which affect organelle function. Several chloroplast retrograde regulatory pathways are described and some of the molecular components that mediate these retrograde pathways have been identified. In contrast, while abundant biochemical and cell biology data indicate the presence of retrograde regulatory pathways from mitochondria to the nucleus in plant, there is little known with respect to the components that mediate these signals. The mitochondrial alternative oxidase (AOX) is widely used as a marker of the retrograde response in plants. It is induced by a variety of stress signals, including interventions that specifically inhibit mitochondrial function. We have used the promoter of the alternative oxidase, linked to the reporter gene luciferase, in a forward genetic screen to identify the components required for alternative oxidase induction under conditions that disrupt mitochondrial function.
This screen has led to the identification of several mutants, for which the associated genes have been identified or are being currently mapped using mapped based cloning and next generation sequencing approaches. The identity of these genes involved in mediating mitochondrial retrograde regulation will be presented, along with insights into whether these genes also affect other mitochondrial retrograde pathways or interact with chloroplast retrograde regulatory pathways.
A new chloroplast retrograde pathway functions in drought, light and abscisic acid signalling in
Arabidopsis
leaves
Estavillo, GM 1
BJ 1
, Pornsiriwong, W 1 , Crisp, PA 1 , Pogson,
1 ARC Centre of Excellence In Plant Energy Biology,
Research School of Biology, Australian National
University, Canberra, Australia
Compartmentation of the cell, which has been critical to the success of Eukaryotes, requires a complex set of subcellular messages including retrograde signals from the chloroplast and mitochondria to the nucleus to regulate gene expression. Although proteins that participate in different signalling cascades in higher plants have been identified, the actual mobile signals and their mechanism of action are debated. Here we demonstrate that a phosphonucleotide, PAP, accumulates in Arabidopsis thaliana in response to drought and light stress and its levels are regulated in the chloroplast by a chloroplastic enzyme, SAL1. PAP moves from chloroplast to the cytosol and nucleus where it inhibits
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the activity of exoribonucleases (XRN). Global expression analyses showed that both SAL1 and XRNs modulate the expression of a very similar subset of genes, including the high light inducible gene expression of APX2 and ELIP2. Equally significant, the PAP pathway activated a novel abscisic acid (ABA) signalling pathway, independent of OST1 and ABI1, restoring germination inhibition, stomatal closure and drought tolerance in ABA-insensitive mutants. Thus, PAP is a retrograde signal regulating RNA metabolism thereby altering nuclear gene expression, during high light and drought stress. Additionally, PAP activates a novel ABAmediated signalling pathway during germination and drought.
Cell–cell communication in the female gametophyte of
Arabidopsis
Gross-Hardt, R 1
Stierhof, Y 1
, Kägi, C 1 , Baumann, N 1 , Nielsen, N 1 ,
1 ZMBP, University of Tübingen, Germany
In flowering plants gametes develop in few-celled haploid structures, termed gametophytes. The female gametophyte of Arabidopsis consists of four distinct cell types. The egg and central cell get fertilized to form the main components of the seed. These gametes are flanked by accessory cells that partially aid in fertilization. We are interested in the mechanisms underlying the specification of the distinct cell types. In a screen for regulators of egg cell fate, we have previously isolated the lachesis mutant which forms supernumerary egg cells. In-depth characterization of the mutant revealed that accessory cells differentiate egg or central cell fate, demonstrating that all cells in the female gametophyte are competent to adopt gametic cell fate. Our data suggest that the mechanism, which suppresses gametic competence in accessory cells is generated in gametic cells, implying that gametic and accessory cells develop in a coordinated manner. Here, we present the isolation and characterization of the FIONA gene, which provides new insights into the molecular basis of this cell–cell communication.
Structure and evolution of megagametophyte in
Podostemaceae
Sehgal, A 1 , Khanduri, P 2 , Tandon, R 2
1 Botany Dept, Miranda House, University of Delhi,
Delhi, India; 2 Dept of Botany, University of Delhi, Delhi,
India
Members of Podostemaceae are aquatic; they grow diageotropically attached to rocks exclusively in running waters of tropical and subtropical rivers of the Old World and New World. The basic body plan of most of the podostemads is thalloid, deviates considerably from the classical root–shoot model typical of angiosperms, and hence is a structural novelty. Plants of this family are
168 characterized by marked structural and developmental reductionism and several unique embryological features such as absence of antipodals, triple fusion and endosperm; and presence of 4-nucleate mature megagametophyte, pseudo-megagametophyte, single fertilization and suspensor haustoria. Of all the embryological features, the highly reduced embryo sac is the most intriguing one as it appears to be closely coupled with the occurrence of single fertilization in this family. The development of the embryo sac has been described by earlier workers as reduced bisporic type wherein the mature embryo sac is 4/5-nucleate/celled. Of the four cells, two differentiate into synergids, third cell into an egg and fourth into a uninucleate central cell whereas the fifth nucleus lies at the chalazal pole. Both the number of gametopytic cells and /or nuclei, and their identity in the mature embryo sac have remained controversial. This embryo sac has been assigned either
Apinagia type-form A, Apinagia type-form B or
Polypleurum type, which are variations of the developmental pattern. Our observations on Dalzellia zeylanica , Hydrobryopsis sessilis, Zeylanidium olivaceum have not only revealed variation in the family but also the presence of a novel type of 3-nucleate/ 3celled mature megagametophyte consisting of two synergids and an egg cell. Interestingly, the single polar nucleus of the central cell degenerates prior to the entry of the pollen tube into the synergid, therefore one of the two male gametes fuses with the egg cell resulting in syngamy whereas the other male gamete eventually degenerates due to the absence of its partner. The phylogenetic position of Podostemaceae with respect to megagametophyte composition and evolution is highly intriguing. Interestingly, the ontogeny of the female gametophyte( in the investigated genera) resembles that of the basal clade but differs significantly in the apoptosis of the central cell and consequently possessing only single fertilization, yet Podostemaceae is presently placed much above the basal clade i.e. in Malpighiales clade in the Eurosids 1 that have monosporic 8-nucleate,
7-celled female gametophyte with double fertilization.
We therefore suggest that the highly reduced, 3-nucleate,
3-celled mature megagametophyte of the investigated genera may not represent the simple module of basal angiosperms but appears to be derived character by a two-step reduction process. First step might have involved the loss of chalazal quartet from bisporic 7celled, 8-nucleate module, highlighting the viewpoint that the micropylar developmental module is a conserved feature. Presumably, the second step witnessed the degeneration of central cell in D. zeylanica and this critical transition led to ‘knocking off’ of double fertilization. Our work highlights the minimalistic nature of megagametophyte and ascertains the occurrence of single fertilization.
Cell–cell communication during fertilization in
Arabidopsis
: a surprising link to disease resistance
Kessler, S 1 , Escobar-Restrepo, J-M
Shimosato-Asano, H 1
1
Grossniklaus, U 1
, Kienath, N 2
1 , Huck, N 1
, Panstruga, R 2
Institute of Plant Biology & Zürich-Basel Plant Science
Center, University of Zürich, Zürich, Switzerland; 2 Max-
Planck Institute for Plant Breeding Research, Köln,
,
,
Germany

Research in our laboratory focuses on the developmental genetics of plant reproduction centering on gametophyte development. Our studies have shown that both genetic and epigenetic mechanisms play a key role in plant reproduction. In this contribution we will focus on the molecular basis of cell–cell interactions during double fertilization. We have isolated a female gametophytic mutant, feronia , which disrupts double fertilization: in feronia mutant embryo sacs the pollen tubes, even if wild-type, are unable to release the sperm cells to effect fertilization (Huck et al., 2003, Development 130: 2149).
This phenotype suggests that the female gametophyte plays a crucial role in pollen tube reception and, thus, controls the behaviour of the male gametophyte. The feronia mutant defines novel signalling processes between the male and female gametophytes in the process of double fertilization. FERONIA was shown to encode a receptor-like kinase of a plant-specific subfamily (Escobar-Restrepo et al. 2007). Interestingly, some interspecific crosses result in phenotypes that are very similar to those observed in the feronia mutant. I will report on the molecular and biochemical characterization of FERONIA and on our search for additional components of this signal transduction process using genetic and biochemical approaches. Our recent attempts to identify novel components of the FERONIA signal transduction pathway have identified surprising links to disease resistance in plants. The evolutionary implications of these findings will be discussed.
An evolutionary–developmental perspective on the angiosperm reproductive syndrome
Friedman, W 1
1 Harvard University, Cambridge, USA
After more than a century of static thought about the diversification of flowering plants, we are in the midst of a fundamental reevaluation of their evolutionary and developmental history. Angiosperms possess a unique suite of reproductive characteristics that includes a highly reduced female gametophyte, a process of double fertilization, and the production of a genetically biparental embryo-nourishing tissue, endosperm. Recent embryological investigations of ancient angiosperm lineages indicate that the first flowering plants are likely to have produced a diploid endosperm derived from a four-nucleate embryo sac. These studies also reveal that the angiosperm female gametophyte is a fundamentally modular entity, and that evolutionary transitions in the number and developmental patterning of these modular subunits directly alter the genetic constitution of endosperm. Thus, evolution of endosperm ploidy, maternal to paternal genomic ratios, levels of heterozygosity, and degrees of genetic conflict (vis à vis its role in interparental conflict and/or parent-offspring conflict) is directly tied to the expression of underlying modular components of the female gametophyte. The key is to understand female gametophyte diversity based on general principals of developmental biology (e.g. modularity, ectopic expression, and heterochrony), and to link this diversity to evolutionary innovations (perhaps even adaptations) associated with changes in endosperm genetics.
Competition within megaspore tetrads
Haig, D 1
1 Dept of Organismic and Evolutionary Biology, Harvard
University, Cambridge, USA
For any particular gene in a megaspore, two of the other three members of its tetrad are non-relatives. Therefore, megaspores are expected to compete to be the haploid mother (mum) of the embryo that develops in a seed.
Usually this competition is restrained by by the suppression of three of the four megaspores in the tetrad by the diploid mother. In most cases, the chalazal megaspore survives to produce a mature female gametophyte within which the egg develops from a micropylar nucleus. In bisporic and tetrasporic gametophytes one or both meiotic divisions is not accompanied by cytokinesis. In consequence, a megaspore other than the chalazal megaspore contributes the micropylar egg apparatus and is thus the mum of the embryo. Bisporic and tetrasporic development may have originated as forms of 'meiotic drive' that conferred a segregation advantage on one of the two alleles in a heterozygous mother.
Plant cell growth and development in microgravity
Kiss, JZ 1 , Millar, KDL 1 , Edelmann, RE 1
1 Miami University, USA
Life has evolved in the constant 1-g conditions of Earth, and gravity has been a ubiquitous force present throughout the evolutionary history of plants. However, the microgravity environment found in orbiting spacecraft provides a useful and unique tool to investigate the mechanics of cellular growth and development in plants. Tropisms, the directed growth in response to external stimuli, are important throughout the life cycle of the plant. We studied the interaction between gravitropism and phototropism in experiments on the
International Space Station (ISS). Because of the interfering effect of the strong gravitropic response, microgravity conditions are needed to effectively study a
‘pure’ phototropic response. The European Modular
Cultivation System (EMCS), which has environmental controls as well as centrifuges for gravity treatments and controls, was used in these studies. In our first series of experiments, we discovered a novel red-light-based phototropic curvature in hypocotyls of seedlings of
Arabidopsis thaliana and that this response is mediated by the phytochromes. In addition to microgravity experiments, we also assayed tropistic curvature in fractional gravity studies (ranging from 0.1 g to 0.8 g ) that were made possible by the on-board centrifuges in the EMCS. We will provide a quantitative analysis of the effects of increasing centrifugal accelerations on phototropic curvature in roots and hypocotyls in order to better understand the interactions between tropisms. Our studies highlight the exciting opportunities to utilize
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research laboratories on the recently completed ISS to study basic questions in plant biology.
Is the oscillating motions of leaves during their unfolding necessary for their straightening?
, Couturier, E 1 , Bastien, R 3 , Heuret, P 2 , Kheffache, R 1
1
Douady, S 1
MSC Laboratory, CNRS-Paris Diderot University,
France; 2 INRA Guyanne; 3 PIAF Laboratory, INRA-
Blaise Pascal University, France
Between the primordial formation to the final maturation of the leaf a long part of the development can happen in a finite volume delimited by older/younger leaves, stipula… In order to keep developing its surface it can enroll or fold (pre-foliation). For folded leaves, we find that the folds appear from an active process of differential growth of the parenchyma around the main veins. The back of these veins expands considerably, rotating the lamina around the vein axis toward the apex.
The longer the development of the leaf happens enclosed in the finite bud volume the older some veins becomes and rotate the lamina, creating a more and more complex leaf shape. However, even during this folded development, it is remarkable that the lamina remains locally flat. This reveals a first very effective regulation.
Only the forced mania fold inside the bud some gaussian curvature can appear, leading to a typical protrusion of the lamina near the sinuses of the leaf once matured.
When the leaf unfolds outside, there is an inversion of the differential growth of the parenchyma around the veins. This opening is often exaggerated and leads to a reverse folding of the leaf, before eventually flattening out. The amplitude of the reverse motion depends a lot on the growth rate of the leaf, and the simple observation of the leaf shape during that stage thus leads to a direct estimation of this growth rate. However, the final expansion of the leaf, even with this exaggerated opening, is not straightforward, and also occurs with many oscillations on an ultradian rhythm (typically 3h).
As these oscillations appear to be quite universal during leaf expansion it is then natural to consider their possible usefulness. Finally, with all these motions, it is even more surprising that the leaves are finally pretty flat and straight. Each oscillation corresponds to a local growth that is not homogeneous, each differential growth leading to a local deformation of the leaf and thus a movement of the rest of it. In order to reach a finally flat state it thus needs (again) strong regulation. However, any regulation requires a way of knowing the distance of the present state to a target state, in order to try to compensate it accordingly. These oscillations could thus be a mechanical way for the leaf of knowing how far it is from being flat.
Mechanical signals in plant development: shaping the shoot apical meristem
Hamant, O 1 , Uyttewaal, M 1
Kwiatkowska, D 2
, Milani, P 1 , Landrein, B 1
, Boudaoud, A 1
1 RDP/LJC, ENS Lyon, France; 2
,
Univeristy of Silesia,
Katowice, Poland
The contribution of the gene regulatory network in morphogenesis involves by definition major changes in structure. The mechanical properties of cells and tissues thus play a crucial role, notably to coordinate growth.
Based on our previous work on the role of mechanical forces in driving microtubule-based cell anisotropy (1, 2) and PIN1 dependent cell polarity (3), we will show here how the microtubule severing protein Katanin is required for growth coordination at the shoot apex. Major morphogenetic defects are observed in the katanin mutant, including the absence of sharp crease in the boundary separating the emerging organ from the meristem, and a curvy meristem surface with bulging cells. These defects can be correlated to the lack of a coherent supracellular microtubule pattern in the meristem, together with heterogeneous cell growth rates and curvatures, as quantified via the replica method
(originally developed by Paul Green). Altogether, our results support a model in which cell–cell communications are impaired, notably because of a slower response to mechanical stimuli. Despite these major shape defects, patterning is only weakly affected, with subtle modifications in gene expression patterns and relatively minor phyllotactic defects. The contribution of mechanics in the robustness of these morphogenetic responses will be discussed. To go beyond the indirect assessment of the mechanical properties of the tissues, we have also developed an atomic force microscopy approach, which allows us to measure the elastic modulus of the outer wall of the meristem epidermis.
Local, sometimes subcellular, heterogeneities can be uncovered using this method leading to a model in which a mechanical zonation can be superimposed to the histological and molecular organization of the shoot apex. (1) Hamant O et al. 2008. (2) Corson F et al. 2009.
(3) Heisler M et al. 2010.
1
A model of plasma membrane flow and cytosis regulation in growing pollen tubes
Chavarría Krauser, A 1 , Du, Y 2
Center for Modelling and Simulation in the Biosciences,
University of Heidelberg, Germany; 2 Heidelberg Institute for Plant Science, University of Heidelberg, Germany
The mechanical properties of growing pollen tube cell walls depend on the chemical composition of the pectinous material. In particular the esterification degree determines rigidity, and thus, the ability to expand.
During growth, pollen tubes have to have a well defined and tightly regulated distribution of cell wall extensibility, or else a stable cap geometry cannot be maintained. Pollen tubes achieve this by influencing the esterification degree through Pectin Methyl Esterases
(PME), which activity is regulated by an inhibitor
(PMEI). Distinct patterns of PME and PMEI are found in pollen tubes. While PME is widely distributed along the flanks of the pollen tube, PMEI is only present at the apical cell wall. To achieve these distinct distributions,
PME and PMEI are subjected to specific cytosis patterns.
The cell wall material, pectin, reaches also the wall by means of exocytosis. It stands to reason that mechanics of growing pollen tubes can only be understood completely if the patterns of endocytosis and exocytosis are also considered. Therefore, we used a theoretical approach to understand these patterns. A model of
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cytosis regulation is developed and simulations presented. We address in particular the question on the minimal assumptions needed to describe the patterns reported recently by Zonia and Munnik (2009). We also discuss several biological implications delivered by the model. The movement of plasma membrane in the tip is described by using concepts of flow and conservation of membrane material. After obtaining the central equations, relations describing the rates of endocytosis and exocytosis are proposed. We find that two cytosis receptors (for exocytosis and endocytosis), which have different recycling rates and activation times, suffice to describe a stable growing tube. The simulations show a very good spatial separation between endocytosis and exocytosis, and separation is shown to depend strongly on exocytic vesicle delivery. The model shows also that most vesicles in the clear zone have to be endocytic, in accordance with the literature. Membrane flow is essential to maintain cell polarity, and bi-directional flow is a natural consequence of the proposed mechanism. For the first time, a model addressing plasma membrane flow and cytosis regulation was posed. Therefore, it represents a missing piece in an integrative model of pollen tube growth, in which cell wall mechanics, hydrodynamic fluxes and regulation mechanisms are combined.
Modeling the mechanics of cell shape generation
Geitmann, A 1 2 , Kroeger, J 3 , Zerzour, R 1
Chebli, Y 1
, Fayant, P
, Girlanda, O 2
,
1 Université de Montréal, Canada; de Montréal, Canada;
2 École Polytéchnique
3 McGill University, Canada
The shape of a plant cell is determined by spatially controlled expansion of the cell wall. To understand the mechanical principles governing this process, we compare the predictions made by computational modeling with microscopic observations. In the rapidly growing pollen tube, at least two opposing mechanical constraints determine the size of this tubular protuberance: For mechanical reasons, a smaller cell can invade the pistillar tissues with greater ease. However, a sufficiently large diameter is required to allow the sperm cells to pass through the tube. In a mechanical model based on finite element techniques we demonstrate how the pollen tube has to control the mechanical properties of its apical cell wall in order to produce a cylinder with the desired size. Intriguingly, the growth speed of the pollen tube typically varies periodically. We developed a mathematical model based on feedback mechanisms involving calcium signaling and exocytosis that is able to reproduce this oscillatory behavior. Importantly, it explains how variations in turgor pressure can influence oscillatory growth even if this parameter is assumed to be stable over time as suggested by micromechanical studies.
Heteroblasty, phase change, and miRNAs – the genetic regulation of shoot maturation
Poethig, SR , Willmann, MR, Li Yang 1
1 Dept of Biology, University of Pennsylvania,
Philadelphia, PA, USA
The vegetative morphology and physiology of the shoot change as it develops. To determine the molecular basis of these changes, we profiled gene expression in shoot apices of different ages and leaves at different positions in early ( FRI flc-3 ) and late-flowering ( FRI FLC ) genotypes of Arabidopsis . Six temporal programs of gene expression in leaves were identified. Three of these programs occur in all leaves (leaf maturation, leaf aging, leaf senescence), two involve changes in the identity of the shoot apex (vegetative phase change, floral induction), and one involves changes throughout the entire shoot (shoot aging). We have focused on the genetic regulation of the vegetative phase change program. Studies of a number of herbaceous species, as well as trees with distinctive juvenile and adult phases
(e.g. Acacia, Eucalyptus ), reveal that this transition is regulated by the microRNAs, miR156/miR157. These miRNAs are expressed at high levels during the juvenile phase and then decline in abundance, leading to the expression of adult traits. The decline in the expression of miR156.miR157 is mediated by a factor or factors produced by leaf primordia. miR156/miR157 act by repressing the expression of SPL transcription factors.
The mechanism of this repression will be discussed.
Compound leaf development in tomato
Ori, N 1
E 1
, Shleizer Burko, S
, Melnik, H 1
1 , Burko, Y 1 , Yanai, O 1 , Shani,
1 Hebrew University, Rehovot, Israel
Compound leaf development in tomato ( Solanum lycopersicum ) provides a sensitive system to explore the establishment of leaf shape, size and complexity. Wildtype tomato leaves are compound, comprising of multiple leaflets, each resembling a simple leaf. Leaf form elaboration relies on the transient maintenance of morphogenetic activity in leaf margins. We have shown that the TCP transcription factor LANCEOLATE (LA), which is regulated by miR319, negatively controls the morphogenetic activity of the leaf margin. Conversely,
KNOXI transcription factors as well as the hormone cytokinin positively regulate indeterminate growth of the leaf margin. Manipulations of the timing of expression and activity of these factors result in changes in the dynamics of leaf growth and maturation, leading to alterations in leaf size and shape. The balance among the antagonistic activities of these and additional factors thus define the window of morphogenesis at the leaf margin.
The NAM/CUC transcription factor GOBLET (GOB) and the hormone auxin affect leaflet localization, separation and growth within the context of a morphogenetically active leaf margin. Genetic analysis suggests that these factors act through parallel, partially redundant, pathways.
Role of the CUC genes in leaf dissection and morphogenesis
1
Laufs, P 1
INRA, Versailles, France
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Leaf margins show various levels of dissection/outgrowth such as lobes or serrations in simple leaves, or leaflets in compound leaves, and contribute to a large extent to inter- and intra-species variation of leaf shape. We previously showed that all types of dissection require the function of the NAM/CUC3 genes. These genes, which code for plant specific transcription factors of the NAC family, were initially identified for their role in organ separation and meristem formation. We showed that they have a conserved expression pattern at the sinus of the marginal outgrowths in a large selection of dissected Eudicot leaves. Furthermore, reducing
NAM/CUC3 activity leads to a reduction in the number and size of the marginal outgrowths. Here, we will present a detailed analysis of the role of the three CUC genes in Arabidopsis leaf serration. By combining genetic analysis with a precise morphometric study of leaf development we will reveal the specific roles of each individual CUC gene during Arabidopsis leaf serration.
We will provide evidences that CUC2 orchestrates a regulatory network involving CUC genes and MIR164a to fine-tune the level of Arabidopsis leaf serration.
Finally, we will present our recent efforts in developing quantitative approaches to link leaf morphogenesis with
CUC activity and further uncovering the associated cellular and molecular events.
How the volume enclosing the leaf during its growth influence its shape
Couturier, E 1 , Courrech du Pont, S 1 , Douady, S 1
1 MSC Laboratory, CNRS-Paris Diderot University,
France
Between the formation of the primordium and the final maturation of the leaf, there can be a long development of the leaf inside a bud, in general an enclosed volume defined by the previous/next leaves, the stipula, scales, etc. One strategy of the leaf to keep growing and expand as much as possible in this enclosed volume is to grow enrolled or folded (pre-foliation). In the case of folded leaves, we found that there is two very different types of folds. The first one, showing on the outside, are located at the main veins of the leaf. One interest is to protect the young leaf from the outside with a nearly continuous frame of protruding veins. The second type of folds are the one that the lamina is forced to make on the inside. It corresponds to a place where only the smallest level of veins connects. What we observed is that the rest of the lamina is filling as much as possible the possible volume left free for the developing leaf. This shows first that there is a limitation of the lamina growth, probably from mechanical contact. The second effect is that, knowing the way the lamina is folded, and the volume limitation for the lamina growth, these two constraints determine the final shape of the leaf. In particular, not only the veins will correspond to the tip of lobes, an the lamina folds will correspond to sinus of the leaf, but also it imposes particular relationships between the size of the lobes and their opening angles. This shows that the shape of lobed leaves can be more a secondary consequence of this development strategy, rather than a primary target. it could reciprocally show that the main aim is to develop the largest possible surface in the allowed volume. It could thus in turn explains the possible variations of the lobes leaf shapes, depending on the available bud volume
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(from young stem to old ones), or depending on the local climatic conditions, the colder regions inducing a longer development inside the bud and thus a more complex contour. This could also explain why there is a predominance of lobed leaves in temperate to cold climate, while the rare lobed leaves observed in tropical climate belongs to colonial plants. Finally, these observations lead to the problem of understanding the development of these folds, with the asymmetric and the contact regulations, as the good way to understand the indirect determination of lobed leaves shapes.
Inter-cell-layer movement of ANGUSTIFOLIA3 coordinates cell proliferation between clonally distinct tissues in leaves
Kawade, K 1 , Horiguchi, G 2 , Tsukaya, H 1
1 The University of Tokyo, Japan;
Japan
2 Rikkyo University,
Coordination of growth between distinct tissues is essential for organ-size and -shape controls. Epidermal and most of mesophyll tissues in leaves are derived from clonally distinct layers of shoot apical meristem called
L1 and L2, respectively. Cell proliferations in these tissues are coordinately regulated by inter-cell-layer communication (reviewed by Savaldi-Goldstein and
Chory 2008), yet the molecular mechanism underlying the coordination is poorly understood.
ANGUSTIFOLIA3 (AN3) is a putative transcriptional co-activator which is involved in positive regulation of cell proliferation in leaf primordia (Horiguchi et al.
2005). Cell proliferation is defective both in epidermal and mesophyll tissues of loss-of-function mutant of AN3 while AN3 transcripts are mainly observed in developing mesophyll tissue but hardly detectable in epidermis
(Horiguchi et al. submitted). These facts raise a possibility that an inter-cell-layer communication for cell proliferation is mediated by AN3 or its downstream pathway, and provide clues that are relevant to understand the regulations of leaf size and shape. In this study, to investigate the potential mobility of AN3, we constructed transgenic plants expressing AN3 fused with
GFP specifically in L1- or L2- lineage in the an3-4 genetic background. PDF1 or CLV1 promoter was used for L1- or L2-specific expression of AN3-GFP, respectively (referred to as an3-4/pPDF1::AN3-GFP and an3-4/pCLV1::AN3-GFP). First, we confirmed the specificities of these promoter activities by GUS reporter assay. We then characterized leaf phenotypes of an3-
4/pPDF1::AN3-GFP and an3-4/pCLV1::AN3-GFP. As a result, leaf size and shape of an3-4/pPDF1::AN3-GFP and an3-4/pCLV1::AN3-GFP was restored to a level comparable to that of wild type. Cell proliferation was also fully rescued both in epidermal and mesophyll tissues of these transgenic lines. Importantly, we observed GFP signal in epidermal and mesophyll tissues of an3-4/pCLV1::AN3-GFP although the activity of
CLV1 promoter in leaf primordia was restricted within developing L2 lineage. This result indicated that AN3 is capable of moving from mesophyll to epidermal tissue for inter-cell-layer coordination of cell proliferation. Our previous results showed that AN3-3xGFP have no mobility between cells and promotes cell proliferation as well as native AN3 does. Thus, to understand the significance of inter-cell-layer movement of AN3, we are

attempting to characterize transgenic plants expressing
AN3 tandemly fused with triple GFP (AN3-3xGFP) in the an3-4 genetic background under the control of AN3 promoter (pAN3::AN3-3xGFP). Recently, we showed that the defect of cell proliferation caused by an3 mutation triggers an intercellular signaling that enhances post-mitotic cell expansion using clone analysis (Kawade et al. in press). Based on this fact together with our findings, we would like to discuss that AN3 should be a key factor for inter-cell-layer coordination of cell proliferation, and post-mitotic cell expansion in leaves.
From genes to shape: modelling the mechanics of plant development
Boudaoud, A 1
1 Ecole Normale Supérieure de Lyon, France
Although many of the molecular aspects of plant development are known in great detail, understanding how genes are translated into shape is still a challenge.
Mechanical forces might be an important player in morphogenesis. How are these forced regulated at the cellular level? How is growth coordinated? Can these forces feedback on cellular behavior? I will present our latest modelling efforts to help answering these questions, focusing on the shoot apical meristem.
Computational models combining gene regulation, hormone signaling, and mechanics can explain developmental processes in the shoot apical meristem
Jonsson, H 1
1 Lund University, Sweden
The shoot apical meristem continues to provide cells to aerial organs throughout the life of a plant. This requires a tight coupling of several mechanisms to keep stem cell maintenence, differentiation, and growth regulated, guiding the development of plant architecture. While recent years have provided us with extensive experimental information on detailed local (cell-based) interactions as well as global (tissue) phenotypes, the connection between local and global behavior is complex. Mathematical descriptions of the systems are needed to provide information on whether the consequences of the local hypotheses lead to the global behavior seen in plants. The emerging research field of computational morphodynamics addresses these questions, iteratively combining dynamic cell-based measurements with mathematical modeling. I will describe how this approach has been used to increase our understanding of how gene regulation, hormone signaling, and mechanical properties can be integrated to get a systems level description of the developmental processes. Especially I will focus on how auxin transport and mechanical anisotropy provide means to drive primordia formation in the shoot apical meristem.
References: Heisler M et al. 2010; Hamant O et al. 2008;
Jönsson H et al. 2006.
Investigating mechanical properties of plant cells and
1 the shoot apex
Smith, RS 1
AL 1
, Nakayama, N 1
, Kochova, P 1 , Weber, A
Lausanne, Switzerland;
1
3
, Scheupback, T 2
, Janka, A
University of Bern, Switzerland;
3
2
, Routier,
University of
University of Fribourg,
Switzerland
Plant development occurs through a carefully controlled interaction between cell differentiation and cell growth.
At the biochemical level genes, proteins and signaling molecules cause cells to differentiate into specialized tissues. This leads to differences in the way cells regulate their growth, often anisotropically, and it is this regulation that ultimately determines the emergent forms seen in plants. Although considerable progress has been made in the understanding of plant patterning events at the biochemical level, little is know about the mechanics of growth and the interaction between biochemical processes that direct it. This is in part due to the fact that the structural aspects of plant tissue are fairly complex.
Plant cells are like small balloons, that exert considerable pressure on the rigid cell wall that contains them. It is this pressure that gives the tissue its structural integrity.
Yet despite the high pressures contained within, plants cells are able to expand, often quite rapidly, to many times their original size as the plant grows. This structural complexity presents a challenge when interpreting experiments aimed at determining cell wall material properties, and their relationship to growth.
Indentation experiments with micro-electro-mechanical systems (MEMS) sensors can be used to measure cell wall material properties in-vivo. However the data comes in the form of force-indentation curves, which do not directly give basic material parameters. The measured values are a combination of turgor, cell wall elasticity, bending stiffness, and viscous properties of the cell wall and cell contents. For this reason, physically-based simulation models of plant cells are required to explore the effects of these components on measured values. The goal is to separate the effects of the individual components by comparing simulation results with force measuring experiments performed under a range of conditions. Since turgor plays such a key role in the structure of plant tissue, another approach to study wall mechanics is to analyze tissue deformation in response to turgor manipulation. This requires a method to precisely quantify cell expansion. Computer software
(MorphoGraphX) developed for the analysis of 3D confocal image stack data can be used to extract the surface geometry of the shoot apex. Cell walls stained with fluorescent markers are then projected onto the the surface and used as landmarks to quantify tissue expansion. By comparing ‘before’ and ‘after’ stacks from turgor manipulation experiments, insight into the material properties of the cell wall can be obtained. The same technique can also be used to quantify tissue expansion due to growth in normal conditions, which can then be directly compared to results from turgor manipulation.
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A general model for the dynamics of stem gravitropism
Bastien, R 1 , Moulia, B 2 , Douady, S 1
1 MSC Laboratory, CNRS-Paris Diderot University,
France; 2 PIAF Laboratory, INRA-Blaise Pascal
University, France
Plants stands vertical only through constant regulation.
Contrary to roots, it is assumed that all along the growing part of the stem there are cells sensitive to the gravity orientation. In primary growth, all the elements of the growing part will react locally to the tilting by differential growth on the upper and lower side, thus creating a local curvature of the stem. If a young stem is tilted it will curve to regain the verticality over the largest possible part. During this motion, our observations first show that most of the stem curves, before finally straightening back vertically and the curvature to localize at the bottom. Here we present a minimal model of this movement that depends only on the curvature of the stem and its orientation toward gravity. The model is doubly local as it supposes that for a local angle at a place along the stem the response will be a local curving. In order to be able to reproduce the observed movements, and in particular the fact that the stem can straighten back even before reaching the vertical, we have also to take into account an autotropic effect, translating the fact that the stem tries to remains straight as much as possible, independently of any orientation. We can show that the final shape is then given by the equilibrium between these two possibly contradictory tendencies. In particular, the relative strength of these two effects defines a typical length, on which the stem can curve, and the ratio of this length with the length of the growing part of the stem gives the final shape of the stem. Interestingly, this is this same ratio that characterizes the motion of the stem itself, during its dynamical response. If the curvature length is large (or the stem short), the stem will at most succeed to reach the vertical. If it is smaller (or the stem longer), then the tip will momentarily over pass the vertical and tilt in the other direction, before coming back stabilizing at the vertical. For even larger values, the stem tip can oscillate more and more times, before stabilizing. The fact that this is the same ratio that characterizes the final shape and its dynamical response thus allows for a quick characterization of the gravitropism of a stem by just observing the final shape. This would allow a quick and easy quantitative screening of gravitropic response. More generally, it is important to realize that the final shape and the dynamics of the response can thus be characterized by the relative strength between the gravitropism and the autotropism. This shows that this effect, the fact the stem tries to remain straight as much as possible independently of any orientation, is an essential one that has not to be forgotten.
Computational morphodynamics reveals the role of cell division in the patterning of
Arabidopsis
sepals
Roeder, A 1 , Chickarmane, V
Koumoutsakos, P 2
1 , Cunha, A
, Meyerowitz, E 1
1 , Tauriello, G 2 ,
1
2
California Institute of Technology, Pasadena, USA;
ETH, Zurich, Switzerland
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The new field of computational morphodynamics has emerged with the goal of elucidating the development of plants in both space and time via the convergence of three strategies: live imaging, image processing, and computational modeling. We have used a computational morphodynamic strategy to address one of the important four-dimensional questions in plant development: how do growth and cell division contribute to the patterning of specialized cell types within an organ? To address this question, we developed the Arabidopsis sepal epidermis as a model system because it has a characteristic pattern of giant cells stretching an average of 360
µ m interspersed between smaller cells that reach as little as
10
µ m. In addition, the sepals are the outermost floral organs, which enclose and protect the developing bud and are consequently accessible for imaging and manipulation. By live imaging we tracked the cell lineage of the early developing sepal over three days and found that very early in development, sepal giant cells enter a specialized endoreduplication cell cycle in which they replicate their DNA and grow, but fail to divide. At the same time, the surrounding cells continue to divide, reducing their area, and thus remain small cells. We built a computational model in which each cell decides randomly when to endoreduplicate with probabilities based on the proportion of endoreduplicated cells in the sepal. Comparing the output of the model with data derived from image processing of living sepals revealed that while the timing of endoreduplication is central to the pattern of giant cells and small cells, it is insufficient to generate the entire diversity of cell sizes. More live imaging showed that the length of the cell cycle is important for cell size because longer cell cycles give the cell more time to grow. A modified model in which both the timing of endoreduplication and the length of the cell cycle are determined randomly is sufficient to reproduce the distribution of cell sizes in the sepal epidermis.
Finally, we further demonstrated that the model could predict the phenotypes of plants with genetically altered cell size patterns. The intertwined and iterative use of modeling combined with biological experimentation allowed us to reach the conclusion that the variability in cell division timing is the mechanism for creating the characteristic and reproducible pattern of cell sizes in the epidermis. The complexity of biology ensures that no model is ever complete, and we are now examining the cellular growth pattern as well as investigating the decision to endoreduplicate at the molecular level.
Modeling plant development with cell complexes in one, two, and three dimensions
Lane, B 1 , Prusinkiewicz, P 1
1 Dept of Computer Science, University of Calgary,
Canada
When constructing models of plant development, it is useful to separate consideration of the geometry of a modeled system from its topology. While some changes to a plant during development, such as the elongation of stems or the enlargement of single cells, are purely geometric changes, many developmental changes, such as branching or cell division, modify the system's topology. Therefore, the choice of how to represent the system's topology is an important one. Traditionally, the modeled topology is that of a graph: components,

whether functional modules, organs, cells, or abstract subdivisions of cells, are connected to their physical or functional neighbours. However, in physical simulation especially, a more appropriate topology is that of a cell complex. In addition to the components of immediate interest, a cell complex models the connections between them as components, along with the connections between the connections, and so on. For instance, a cell complex model of a three-dimensional multicellular organism models not only the three-dimensional cells, but the twodimensional cell walls connecting them, the onedimensional junctions connecting the cell walls, and the zero-dimensional points where cell wall junctions meet.
Models of the development of filamentous or branching structures have a one-dimensional connectivity, even if their components are three-dimensional. L-systems provide a well-defined mathematical framework for creating these models, using a graph topology; they can also be used to model these filamentous or branching structures as one-dimensional cell complexes. Significant progress has also been made in extending the ideas of Lsystems to the modeling of systems with twodimensional connectivity, such as cellular layers. The formalisms developed so far, such as vertex-vertex systems, are not (yet) as elegant as L-systems, but they too can represent these systems as two-dimensional cell complexes. However, formal description and modeling of the development of systems with three-dimensional connectivity (such as three-dimensional multicellular structures) has largely remained an open problem.
Previous extensions of L-systems to three dimensions rely on developmental rules that are difficult to specify and do not fully capture the topology of the resulting structure. A new modeling framework for threedimensional cell complexes has been developed. This new framework avoids some of the problems of previous models of three-dimensional plant development by explicitly modeling the entire cell complex. While it is not a direct extension of L-systems to three dimensions, it does extend the idea of the graph rotation system which is so useful in vertex-vertex systems. This provides a useful mechanism for navigating the cell complex structure in a local manner. In this presentation, the ideas behind modeling with cell complexes will be introduced, and some examples of models in one and two dimensions will be shown. The new three-dimensional framework will then be described and demonstrated with a threedimensional model of the shoot apex of the moss
Physcomitrella patens .
DigR : how to model root system in its environment?
1 - the model
Rey, H 1 , Barczi, J-F 1 , Jourdan, C 2
1 CIRAD-AMAP, Montpellier, France; 3 CIRAD-
Eco&Sols, Montpellier, France
Many models already exist through literature dealing with root system representation, among which pure structure models such as Root Typ (Pagès 2004),
SimRoot (Lynch 1997), AmapSim (Jourdan 1997); diffusion PDE models (Bastian 2008; Bonneu 2009) and structure/function that are rather scarce and recent
(Dupuy 2010)may be aroused. Nevertheless in these studies, root architecture modeling was not carried out at organ level including environmental influence and not designed for integration into a whole plant characterization. We propose here a multidisciplinary study on root system from field observations, architectural analysis, formal and mathematical modeling and finally software simulation. Each speciality is individually investigated through an integrative and coherent approach that leads to a generic model (DigR) and its software simulator that is designed for further integration into a global structure/function plant model.
DigR model is based on three main key points: (i) independent root type identification (ii) architectural analysis and modeling of root system at plant level; (iii) root architecture setup indexed on root length.
Architecture analysis (Barthelemy 2007) applied to root system (Atger 1994) leads to root type organisation for each species. Roots belonging to a particular type share dynamical and morphological characteristics. Root architectural setup consists in topological features as apical growth, lateral branching, senescence and death, and geometrical features as secondary growth and axes spatial positioning. These features are modeled in DigR through 23 parameters whose values can evolve as a function of length position along the root axes for each root type. Topology rules apical growth speed, delayed growth, death and self pruning probabilities. Branching is characterized by spacing and mixture of lateral root types. Geometry rules root diameter increase, branching and growth directions (including local deviations and global reorientation). DigR simulator provides a user interface to input parameter values specific to each species. It is integrated into the Xplo environment
(Taugourdeau 2010). Its internal multi-scale memory representation is ready for dynamical 3D visualization, statistical analysis and saving to standard formats
(MTG(Godin 2007), Obj,). DigR is simulated in a quasiparallel computing algorithm and may be used either as a standalone application or integrated in other simulation platforms. This will allow further implementation of functional – structural interactions during growth simulation. The software is distributed under free LGPL license and is dedicated both to biologists and modelers.
Shown applications (fig. 1) mimic the diversity of root systems and emphasize the genericity of the model according to different sets of parameter values. Examples
(fig. 2) prove that additional knowledge may be plugged to DigR to simulate root plasticity facing environmental constraints. Further work will be carried out to apply
DigR to various species and to connect DigR to biophysical soil models (Gérard 2008; Zhang et al.
2002); to aerial part models (Barczi 2008); to ecophysiological models (Mathieu 2009, Bornhoffen
2007); and finally to mix this pure descriptive model to a
PDE model that handles fine root diffuse modelling
(Bonneu 2009).
The role of protein transport systems in chloroplast
1 differentiation
Soll, J 1
Ludwig-Maximilians-University, Germany
175

In flowering plants chloroplasts differentiate from proplastids, which are structurally and functionally very reduced organelles in comparison to the photosynthetically active ones. The development of a photosynthetically active chloroplast requires the synthesis and import of a myriad of proteins from the
Cytosol and their assembly into functionally active units like the photo systems. Since proplastids contain no or very little internal membrane system thylakoid biogenesis requires also the synthesis of vast amvients of polar lipids to form membrane bilayer prerequisite for protein insertion and complex assembly. Early in chloroplast differentiation the thylakoids are at least in part formed by invagination of the inner envelope membrane. Later, a vesicle transport system seems to take over to replenish thylakoids with pigments, lipids or even proteins. Due to their endosymbiotic origin, plastids still contain their own genome and transcriptiontranslation machinery. All photosynthetic complexes including Rubisco are of dual genetic origin, i.e. subunits are encoded for on the nuclear genome or the plastome.
Gen expression, translation, transport and assembly of chloroplast proteins must therefore be coordinated in time, space and quantity to guarantee a sustainable, resource optimized organelle biogenesis. We have identified numerous subunits of the chloroplast protein translocation machinery, named Toc- and Tic-complex.
Developmental and organ specific expression of Toc and
Tic subunits or their isoforms indicates a rather complex regulatory network in the differential assembly of the translocon units, while the major channel forming proteins Toc75 and Tic110 are required at all plastid developmental stages. Only a few translocon subunits present in primary plastids can also be detected on secondary or complex plastids. Evidence exists for homologues of Tic75, Tic22 and Tic20 even in
Apicoplasts of Plasmodium and Toxoplasma , but also in the ancestors of chloroplasts the cyanobacterial. The evolution of the protein import apparatus was a key event in the endosymbiotic process.
Subcellular and subplastidial proteomics to study intracellular and intraplastidial trafficking of proteins
Rolland, N
D 1
1 , Salvi, D
, Masselon, C 2
1 , Brugière, S 2 , Seigneurin-Berny,
, Ferro, M 2
1 Laboratoire de Physiologie Cellulaire Végétale, CNRS,
CEA, INRA, Université Joseph Fourier, Grenoble,
2 France; Laboratoire d’Etude de la Dynamique des
Protéomes, CEA, INSERM, Université Joseph Fourier,
Grenoble, France
Recent advances in the proteomic field have allowed high throughput experiments to be conducted on chloroplast samples. Many proteomic investigations have focused either on the whole chloroplast fractions or on independent suplastidial fractions. However, these previous studies raised the question of the accurate localization of many proteins that were identified in different suplastidial compartments. We recently went a step further into the knowledge of A. thaliana chloroplast proteins with regards to their accurate localization within the chloroplast. To achieve this goal, we first obtained highly pure subfractions of envelope, stroma and thylakoids and evaluated their cross-contaminations
176 using biochemical methods. We then performed a comprehensive analysis of the Arabidopsis thaliana chloroplast proteome starting from the whole chloroplast and its three main compartments. Then, we assessed the partitioning of each identified protein in the three abovecited compartments using a semi-quantitative proteomic approach and also performed a curated information on envelope proteins (Ferro et al. 2010). An in depth investigation of the proteins identified within the purified envelope fraction allowed new insights over this subplastidial compartment to be revealed (Joyard et al.,
2009; Joyard et al., 2010). During the course of this project, we also generated an important tool to investigate the dynamics of the chloroplast proteome at the scale of the whole organelle. This yet unique tool is the first database based on the accurate mass and time tags (AMT) strategy dedicated to plants: the chloroplast
AMT database AT-CHLORO
(http://www.grenoble.prabi.fr/at_chloro/). This proteomic database was generated in such a way that it can be used for quantitative studies (e.g. comparisons of mutants, impact of adverse growth conditions...). This AMT-based strategy is now used to characterize mechanisms that regulate protein trafficking between the cytosol and the chloroplast, to identify chloroplast proteins that are subjected to these regulatory mechanisms before being imported or during their import into the chloroplast and to understand the physiological significance of these mechanisms. (Joyard J et al. 2009, 2010; Ferro M et al.
2010).
Biogenesis of thylakoid membranes in higher-plant chloroplasts
Charuvi, D 1,2
Z 1 , Reich, Z 2
, Nevo, R 2 , Shimoni, E 3 , Kiss, V 2 , Adam,
1 The Robert H. Smith Institute of Plant Sciences and
Genetics In Agriculture, Israel; 2 Dept of Biological
Chemistry, Weizmann Institute of Science, Rehovot
76100, Israel; 3 Electron Microscopy Unit, Weizmann
Institute of Science, Rehovot 76100, Israel
The photosynthetic machinery is hosted within specialized paired membranes called thylakoids, which form elaborate 3D lamellar networks. The process in which undifferentiated plastids (proplastids) gain their thylakoid networks and mature into functional chloroplasts is poorly characterized. In higher plants, this process takes place within the vegetative shoot apex, which consists of the shoot apical meristem (SAM) and leaf primordia. It has been believed that plastids in the
SAM lack chlorophyll-binding proteins and thylakoid membranes, while the leaf primordia, only tens of microns away, already possess functional chloroplasts.
Using TEM and STEM tomography, we probed plastids distributed over the shoot apex of Arabidopsis . Notably, we found that plastids in the L1 and L3 layers of the
SAM, which do not contribute to the formation of the major photosynthetic tissue of the plant, possess fairly developed thylakoid networks. The presence of thylakoid membranes in the L1 layer is intriguing, as this layer gives rise to the leaf epidermis, which, aside from stomatal guard cells, does not contain functional chloroplasts. This suggests that most plastids of the L1 layer lose their thylakoids whilst they are in the leaves. In contrast to plastids in the L1 and L3 layers, those found

in the stem cell-containing region (central zone) of the
L2 layer, which generates the primary photosynthetic tissue of the leaf, contain only vesicles and short membrane fragments. These plastids acquire thylakoid membranes once cells reach the periphery of this layer, from which they are recruited to the leaf primordia. The presence of a sharp developmental gradient in the L2 layer is therefore highlighted. Analysis of plastids in leaf primordia indicates that thylakoid network development does not end in young or even relatively mature primordial leaves, but likely continues throughout the course of leaf development. The presence or absence of thylakoid networks was found to correlate with the presence or lack of chlorophyll-binding proteins, as evident from chlorophyll fluorescence and immuno-EM studies.
From proplastids to chloroplasts in flowering plants
Small, I 1
1 ARC Centre of Excellence in Plant Energy Biology,
University of Western Australia, Perth, Australia
As (in Arabidopsis , at least) the organellar genomes are intractable to most genetic approaches such as mutagenesis or genetic transformation, it is difficult to study the involvement of organelle-encoded proteins in developmental processes genetically. However, as we learn more about the gene expression machinery in plant organelles, it is becoming increasingly clear that there is a wide range of nuclearly encoded factors involved.
Many of these are implicated in essential RNA processing events, without which specific organellar genes fail to be expressed. These nuclearly-encoded factors are amenable to genetic approaches, and mutants lacking these factors can be used as surrogates for organellar mutants that would be impossible to obtain.
We have established a collection of RNA processing mutants that between them display defects in expression of a wide range of plastid and mitochondrial genes. Of particular interest to this symposium are the mutants affected in expression of components of the chloroplast gene expression machinery itself, notably several mutants with depressed levels of or defective RNA polymerase subunits. Mutants such as clb19, ys1, otp70 and dot4 share characteristic plastid gene expression patterns due to a failure to edit or splice rpoA, rpoB or rpoC1 transcripts encoding RNA polymerase subunits.
They also share some developmental characteristics such as a delay in greening. Such mutants are useful for understanding the extent to which plant development depends upon organelle gene expression. In particular, via this route we can examine the extent to which the characteristic changes in nuclear gene expression that usually occur in concert with chloroplast development require correctly functioning plastid gene expression.
The cytoskeleton and the peroxisomal-targeted
SNOWY COTYLEDON 3 protein are required for chloroplast development in
Arabidopsis
Albrecht, V 1 , Badger, M 1 , Apel, K 2 , Pogson, B 1
1
2
Australian National University, Canberra, Australia;
Boyce Thompson Institute for Plant Research, USA
A critical process in plant growth and seedling establishment is the development of the chloroplast. A mutant group in Arabidopsis has been identified with impaired chloroplast development which has been named as SNOWY COTYLEDON (sco). The sco3 mutation was in a member of a largely uncharacterised protein family which is unique to the plant kingdom. The sco3-1 mutation alters chloroplast morphology and development, which results in membranous protrusions from the chloroplast, reduced chlorophyll accumulation, impaired thylakoid formation and photosynthesis in seedlings but also in photoinhibition under extreme CO
2 concentrations in mature leaves. Yet, there were no apparent changes to chloroplast transcription, import of
GFP or protein complex assembly that explain the disruption to chloroplast biogenesis. Indeed, localization studies showed that SCO3 is actually targeted to another organelle; specifically to the periphery of peroxisomes.
However, impaired chloroplast development cannot be attributed to perturbed peroxisomal metabolic processes involving germination, fatty acid beta-oxidation or photorespiration; though there are novel changes in low and high CO sensitivity in seedlings and young true
2 leaves. Significantly, in sco3-1 there are changes to the cytoskeleton and application of amiprofosmethyl, a microtubule inhibitor, has a similar effect on chloroplast biogenesis as observed in sco3-1. In fact, the localization of the SCO3 protein to the periphery of the peroxisomes was shown to be dependent on a functional microtubule cytoskeleton. In conclusion, a previously uncharacterised protein, SCO3, is required for chloroplast biogenesis in seedlings and the sco3-1 mutation and microtubule inhibitors reveal a requirement for the cytoskeleton and peroxisomes in chloroplast biogenesis.
Development of structural and biochemical chloroplast dimorphism in Kranz versus non-Kranz
3
C
4
chenopods
Koteyeva, NK
Russia;
1 , Voznesenskaya, EV 1 , Berry, JO
Washington State University, Pullman, USA
2 ,
1
Edwards, GE 3
VL Komarov Botanical Institute of RAS, St Petersburg,
2 State University of New York, Buffalo, USA;
C
4
photosynthesis requires spatial separation of phases of carbon assimilation which is achieved by development of mesophyll (M) and bundle sheath (BS) (dual-cell Kranz anatomy) or different cytoplasmic domains in individual chlorenchyma cells (single-cell non-Kranz anatomy).
Both systems require biochemically dimorphic chloroplasts with one type supporting the carboxylation phase of the C
4
cycle through expression of PPDK, and the other fixation of the CO
2 decarboxylases, by Rubisco in the C
, generated by C
4
3
cycle. There is also structural dimorphism in grana development, depending on the biochemical C
4
decarboxylase subtype. To establish this complex system structurally and biochemically, C
4
photosynthesis requires a highly coordinated expression of many genes during leaf development. While transcriptional control can contribute to differentiation of chloroplasts in the Kranz system, single-cell C
4
depends on post-transcriptional regulation through intracellular targeting of mRNA or polypeptides from photosynthetic genes. We studied development of structural and biochemical features in
177

representatives of four structural types of C
4
in subfamily
Suaedoideae (Chenopodiaceae): Suaeda taxifolia (Salsina
Kranz), S. eltonica (Schoberia Kranz), Bienertia sinuspersici (non-Kranz) and Suaeda aralocaspica (non-
Kranz). All are NAD-ME biochemical type with structurally dimorphic chloroplasts (well developed grana in chloroplasts having Rubisco, and less grana in chloroplasts supporting the carboxylation phase of the C
4 cycle). Light microscopy and TEM were used to study structural transitions during chloroplast differentiation.
Immunocytochemistry and in situ mRNA hybridization were used to show the temporal and spatial pattern of
Rubisco LSU gene expression. In both Kranz types, during leaf development a structural transition occurs from monomorphic chloroplasts in the two cell types, followed by BS and M cells vacuolization and selective positioning of organelles in BS cells, and finally establishment of BS and M chloroplast dimorphism and mitochondrial differentiation. In the non-Kranz types structural dimorphism occurred after development of the two chloroplast-containing cytoplasmic domains.
Rubisco LSU transcription and translation activities in chlorenchyma cells increased basipetally in parallel to the formation of Kranz anatomy, or to formation of two cytoplasmic domains in single-celled Chenopods. In both
Kranz type species, rbc L mRNA, as well as the corresponding proteins, were located preferentially in chloroplasts of BS cells beginning from the earliest stages of leaf development and prior to chloroplast structural differentiation. In contrast, young leaves of both non-Kranz C
4
species have a clearly defined developmental stage in which a C
3
-like pattern of rbc L mRNA and protein distribution is maintained in both chloroplasts during formation of the two cytoplasmic domains. Selective partitioning of Rubisco to one chloroplast type occurs later and correlates with establishment of chloroplast structural dimorphism. In summary, the Rubisco LSU transcript and peptide accumulation patterns in the Kranz type species suggests control by transcription (or mRNA stability) occurs very early in development before structural dimorphism and positioning of BS chloroplasts; whereas, in the singlecelled C
4
species post-transcriptional targeting occurs only after development of the two cytoplasmic domains and chloroplast structural differentiation. The results indicate that different ontogenetic programs of dimorphic chloroplasts development evolved independently in
Kranz and single-cell C
4
species in subfamily
Suaedoideae.
Adaptation to changing climate in
Arabidopsis thaliana
Schmitt, J 1 , Wilczek, A 2 , Fournier-Level, A 1
1 Brown University, USA; 2 Deep Springs College, USA
If species cannot evolve fast enough to adapt to rapid environmental change, they may be unable to persist.
However, evidence for such adaptational lag is scarce.
We tested for lagging adaptation to warming climate in
178 the annual weed Arabidopsis thaliana in common garden experiments in four sites across the species’ native
European range. Genotypes originating in climates similar to the site of planting had high relative fitness in each site, providing direct evidence for adaptation to climate in this model species. However, genotypes originating in climates historically warmer than the site of planting had higher relative fitness than native genotypes in every site. This result suggests that adaptive evolution may not be fast enough to track future rapid climate change, even in an annual plant. Genome-wide association analysis shows that different loci contribute to fitness in different sites; thus the genetic basis of adaptation to climate may vary across a species range.
Clade-based evo-devo: fruit shape diversification in
Paysonia
(Brassicaceae)
Doust, A 1 , Borrone, J 1 , Borja, G 1
1 Oklahoma State University, USA
Studies of the evolution of development (evo-devo) can be performed at multiple scales, with examplars sampled according to their level of genetic resources, interesting morphology, or phylogenetic position. Within these possibilities, an approach that is seldom attempted is to examine all members of a clade and ask how patterns of gene action change to give the morphological diversity within that clade. Such an approach closely follows the evolutionary history of a group as all extant species are examined. It is difficult to find a group of an appropriate size with interesting morphological diversity on which to attempt such a study, yet we are developing such a study group in the small Brassicaceae genus, Paysonia .
Paysonia has eight species that are diploid and diverse in flower color and fruit morphology. In particular, fruits vary from globose to either medially or laterally flattened, changes which are found repeatedly throughout the family. The genus is found in Texas, Oklahoma,
Tennessee and Alabama, with the four species in
Tennessee being particularly closely related, as evidenced by gene flow between species and incomplete sorting of gene alleles. The species are morphologically discrete, on both flower and fruit characters, as well as habitat and location. We are using a combined phylogenetic and population genetic approach to understand how population-level processes of selection and drift make an impact on the evolution of new species and new morphologies. Six of the eight species can be hybridized, allowing us to create multiple mapping populations where quantitative trait loci (QTL) can be mapped and candidate genes for divergent morphologies identified. To date we have focused on a single cross between two species with divergent flower color and fruit morphology, where we have established a mapping population and are examining candidate genes including
PEAPOD1 and 2, CYP78A9, and GORDITA. Leaf and fruit transcriptome analyses have identified a number of other genes that differ between the two species. Genetic mapping and QTL analyses are ongoing, and will eventually include all of the six crossable species in a nested association mapping approach. We anticipate that this close marriage of phyletic, population genetic, and developmental genetic approaches will yield fresh insight into patterns and processes underlying morphological diversification.

Homoplasy and the genetics of floral novelty in the
Zingiberales
Specht, C 1 , Bartlett, M 1 , Almeida, AM 1
1 University of California, Berkeley, USA
The natural diversity of floral forms within the
Zingiberales, including the development of novel organ structures and shifts in floral symmetry, enables us to leverage the extensive knowledge gained from floral development research in model systems to investigate the developmental genetic processes involved in the evolution of functional phenotypes. Utilizing a candidate gene family approach combined with genomic, comparative transcriptome and in situ expression data, we are investigating the convergent evolution of pollination syndromes across the order. Our goal is to obtain a full understanding of which genes and gene networks are responsible for major developmental changes associated with the evolution of pollinator preferences and shifts in diversification rates across major lineages.
Co-option of transposons to host genes: developmental implications of a novel evolutionary process
Joly-Lopez, Z 1 , Juretic, N 1 , Bureau, T 1
1 McGill University, Canada
Transposable elements (TEs) can be considered ‘selfish’ because they propagate within host genomes without necessarily providing a selective advantage, and can even be detrimental to host gene function. Nevertheless, there are a growing number of reported genes that have originated from TEs in a process called TE domestication. One example is the novel family of domesticated transposons in plants, MUSTANG (MUG), initially discovered using a bioinformatic approach. Our objective is to use an experimental approach to characterize two genes from this family, MUG1 and
MUG2, in the model plant Arabidopsis thaliana and address the question: what is the functional role of MUG in plants? MUG is derived from the TE superfamily of
Mutator-like elements (MULEs) and is present in all the major angiosperm groups, but not in gymnosperms, suggesting that this family might have been domesticated near the rise of flowering plants. All MUGs contain three conserved domains, MuDR, MULE, and a SWIM. These conserved domains and canonical residues are also found in other plant domesticated MULEs FAR1 and FHY3, which are transcription factors (TFs) involved in phytochrome A-mediated response to far-red light. This similarity might suggest that MUG also functions as a
TF. To address this question, a reverse genetics approach was used, where T-DNA mutagenized Arabidopsis plants were grown and selected to generate homozygous lines for mug1 and mug2. Because no obvious phenotype was observed in plants with single mutant alleles, mug1 and mug2 plants were crossed to obtain the double mutant mug1mug2, which showed a strong pleiotropic phenotype compared to the wild-type progenitor ecotype
(Columbia). An extensive phenotypic analysis of mug1, mug2 and double mutant lines was then performed by examining various fitness characteristics that were broad enough to cover all developmental stages. Parameters such as developmental timing, primary inflorescence height, flowering time, and seed set were measured. The results revealed a strong detrimental effect on fitness in mug1mug2 plants, observed as early as the first developmental stages. We also found more subtle but significant differences in the single mutants that were otherwise indistinguishable. In addition, mug1 may have a broader or more important functional role than mug2, since single mutants mug1 show a stronger mutant phenotype than mug2. Microarray analysis using ATH1
Genome Array (Affymetrix) was performed using RNA isolated from mutant seedlings and flowers, and the expression profile was compared to a wild-type Col-0.
Results showed changes in many genes implicated in metabolism, such as electron transport, photorespiration and other enzymatic activities. This led us to suspect that
MUG genes may be expressed in key metabolic organelles such as chloroplast, mitochondria and peroxisome. By studying the MUG gene family in
Arabidopsis thaliana , we hope to shed light on the functional importance of non-coding DNA regions. We also hope to elucidate the process of TE domestication which, given that genomes are comprised in largely of
TEs, is an important potential contributor to novel coding regions in the evolution of host genes.
Mechanisms of evolutionary developmental process acting within species: a key step in understanding between species evo-devo differences
Zanis, M 1
1 Purdue University, USA
Linking genetic diversity to naturally occurring variation in complex phenotypic traits is one of the most important objectives in modern biology. This goal is especially true for evolutionary developmental biology, where we have an extremely rich knowledge of the molecular basis of complex developmental phenotypes and strong evidence for their role in between species variation and even reproductive isolation. However, our knowledge is lacking when we ask 'Do loci characterized through knockout studies in model systems show molecular genetic variation in natural populations that associate with naturally occurring phenotypic variation?' This is an essential missing link in evolutionary developmental biology because we must understand the mechanistic evolutionary process acting within species if we hope to characterize the adaptive (or non-adaptive) developmental differences among species. A large number of genes that govern plant morphogenesis have been functionally characterized in model and emerging model plant systems. These identified genes and genetic pathways clearly regulate complex phenotypic traits which are important for the fitness of an organism, yet we have little understanding of the molecular evolution of these developmental genes and the phenotypes they control in natural populations, especially at interspecific
(between closely related species) and intraspecifc (within and between natural populations) levels. We are leveraging knowledge from developmental genetic studies in model grasses, Zea and Oryza , and using molecular evolutionary analyses and association mapping to examine the evolution of inter- and intraspecific variation in inflorescence architecture in the freshwater
179

aquatic grass, Zizania sp. (North American Wild Rice).
Our population genetic analyses of Zizania show that the populations are highly structured with little gene flow between populations. Isolated populations can lead to several interesting population-level phenomena including local adaptation. Our molecular evolutionary analyses of several developmental loci have revealed unique patterns of duplication unique to the Zizania lineage as well as differential intra-specific rates of evolution between developmental loci paralogs. We show how these patterns of molecular evolution in developmental loci correlate with several identified inflorescence architectural traits that resemble classic Oryza (e.g.,
LAX)and/or maize mutants (e.g. ramosa2).
Union of floral whorls in species of the Neotropical
Galipeinae (Rutaceae) and its implications for the evolution and pollination of the group
, Pansarin, ER 2 , Pirani, JR 1
1
El Ottra, JHL 1
Dept of Botany, University of São Paulo, São Paulo,
Brazil; 2 Dept of Comparative Biology, University of São
Paulo, Ribeirão Preto, Brazil
Most genera of the neotropical subtribe Galipeinae (tribe
Galipeeae, Rutoideae) present several forms and degrees of fusion between the flower organs, including the union of petals into an apparently sympetalous corolla, the joining of the stamens inter se and to the corolla, and the partial to complete connation of carpels. Though these and other floral traits are used in the circumscription of species in Galipeinae, few studies have demonstrated in detail in which extend these unions occur for most of its species. To elucidate these anatomic conditions, and to establish accurate homologies for evolutionary studies, a morpho-anatomic study of the flowers of eight genera of
Galipeinae was carried out. Using cross-sections of pickled material and SEM observations, union of the flower segments were investigated. In the gynoecium analysis is noteworthy the great variability presented in the extent of fusion of the carpels’ margins. Galipea exhibits carpels connate axially and laterally in most of its length, while other genera like Conchocarpus,
Erythrochiton, Ravenia and Almeidea present a greater degree of apocarpy, though the five carpels still unite axially in its basal part and distally, to form a single style. The occurrence of nearly apocarpous gynoecia and the postgenital fusion of its apical or subapical parts is traditionaly associated with the formation of a compitum at anthesis, which would provide advantages of a syncarpous to the apocarpous gynoecium. The formation of the floral tube is also noteworthy. It is formed, in most of the species, by the agency of the filaments which alternate with the petals. In Conchocarpus heterophyllus,
C. minutifloru s and in some specimes of Almeidea rubra , the floral tube is formed by the marginal cohesion of petals and adhesion of petals and filaments through the close intertwining of trichomes – a case of pseudosympetaly. In Galipea and C. macrophyllus , even though pseudosympetaly occurs distally, the floral tube is formed through adnation of filaments to the corolla in its basal part. Only Erythrochiton brasiliensis and Ravenia infelix present a floral tube formed solely by marginal fusion of floral whorls. In the first species it is formed through adnation of filaments to the petals, and in the latter species only through connation of petals, although
180 adhesion of filaments to the corolla also occurs through the intertwining of trichomes. Tubular flowers are present in the majority of Galipeinae species, but this character is uncommon among the rest of the family.
Therefore, it would be expected that flowers of
Galipeinae offer nectar as reward, being pollinated by nectar-seeking animals. This has been demonstrated in some studies about floral biology in this subtribe.
Galipea jasminiflora is pollinated by species of
Lepidoptera and E. brasiliensis is pollinated by hummingbirds. In addition, we have observed pollination by butterflies and hummingbirds in A. rubra . Further investigation of these floral features in more genera of subtribe Galipeinae is necessary in order to allow accurate interpretation of the floral evolution in the light of the available phylogeny of the group.
Evolution and development of secondary growth and wood formation
Groover, A 1 , Du, J 2 , Miura, E 1 , Robischon, M 3
1 US Forest Service, USA;
Zhejiang University, China;
2 College of Life Science,
3 Institut für Forstbotanik und Baumphysiologie, Albert-Ludwigs-Universität
Freiburg, Freiburg, Germany
Vascular cambia and associated secondary growth represent major innovations in vascular plant evolution, and enabled plant forms ranging from tropical lianas to massive forest trees. Major evolutionary events leading to secondary growth as exemplified by extant forest trees can be inferred based on phylogenetic relationships among woody plants, as well as recent results of gene expression and function studies in the model tree genus,
Populus. We present functional characterizations that
Class I KNOX and Class III HD ZIP transcription factors, which have been co-opted from the shoot apical meristem to function in regulating the development of the cambium and secondary vascular tissues. Class I KNOX transcription factors ARBORKNOX1 and 2 regulate specific suites of gene associated with cambial daughter cell differentiation, in part through affecting auxin transport in stems. Class III HD ZIP popREVOLUTA affects patterning of stems undergoing secondary growth, and misexpression results in formation of ectopic cambia.
Class III HD ZIP popCORONA, on the other hand, primarily affects cell differentiation. Our results along with other recent results from other labs are beginning to reveal the specific genes and mechanisms regulating secondary growth, which also speak to evolutionary processes leading to the fundamental plant developmental process.
Habit shifts within Balsaminaceae, with special emphasis on the origin of woodiness and the underlying genetic background mechanism
Smets, E 1 , Melzer, S 2 , Janssens, S 2 , Eeckhout, S 2 , Lens,
1
F 1
University, Leiden, The Netherlands; Laboratory of
Plant Systematics, K.U.Leuven, Leuven, Belgium
National Herbarium of The Netherlands, Leiden
2
The family Balsaminaceae consists of only two genera, which are characterized by a completely different species

composition. While the genus Hydrocera is monospecific, its sister genus Impatiens comprises over
1000 species and belongs to the top twenty most speciesrich genera within angiosperms. Despite the very large number of species, the overall vegetative morphology of
Impatiens is fairly uniform, always having glandular toothed leaves and an herbaceous semi-succulent stem.
On the other hand, thorough anatomical observations of the stem of 35 species, belonging to Hydrocera and all major subclades of Impatiens , reveal much more variation in the development of secondary tissues than previously anticipated. The Impatiens species investigated show a wide range of wood production, ranging from only few cells limited to the intrafascicular bundles (true herbaceous species) towards a complete wood cylinder consisting of about 100 cell layers in thickness (truly woody species). Furthermore, our observations illustrate that secondary growth in
Hydrocera triflora is confined to the corners of the stem.
When plotting the different habit types on a molecular framework, it is evident that most basal Impatiens species do not produce significant amounts of wood, whereas woodier species are present among the more recently diversified lineages of the molecular topology, suggesting that wood formation has been secondary derived. The derived origin of wood formation in
Impatiens is supported by the so-called paedomorphic wood features: all the species observed have declining of flat length-on-age curves for vessel elements, wide gaping intervessel pitting and they lack rays. The microscopic wood structure of Hydrocera strongly resembles that of Impatiens , but the presence of
(paedomorphic) rays and alternate intervessel pitting towards the cambium makes the wood structure less paedomorphic than in Impatiens. The development of secondary wood in Impatiens is linked to our recent genetic findings showing that downregulation of two
MADS box genes in the herbaceous Arabidopsis thaliana wild-type leads to a woody shrub habit.
Is there really a PTM (primary thickening meristem) in monocots?
Menezes, NL de 1 , Elbl, PM
Silva, C 1 , Oliveira Costa, G 1
1 , Cury, G 1 , Gonçalves da
, Sasaki, KLM 1
1 Universidade de São Paulo, Brazil
Primary tissues in monocots are the same as in gymnosperms and in other angiosperms: the epidermis
(originated from protodermis), the cortex (whose innermost layer is the endodermis), which is originated from the ground meristem, the pericycle (outermost layer of the vascular system) and primary xylem and phloem
(originated from the procambium). Since the nineteenth century, researchers have recognized the perycicle as the tissue generating roots, a reason why it was named the dyctiogenic layer. In the last 50 years, a group of researchers established that the primary thickening in stems of monocots, mainly rhizomes, corms and bulbs, occurred by the activity of a lateral meristem, which they called the Primary Thickening Meristem (PTM).
According to these authors, the PTM would be the generating layer of the roots. More recently a group of researchers have shown in stems of monocots an important characteristic described only for roots: an endodermis with meristematic activity forming part of the cortex or even the entire cortex. It has been found that the pericycle and the endodermis both in meristematic phases simulate a single meristem, for they are adjacent to each other. However, both tissues have distinct functions: endodermis initials have only a centrifugal function, forming the cortex (or part of it), while the meristematic pericycle have centripetal function, forming the vascular system, although originating also adventitious roots in the stem and lateral roots in roots.
Recent research studies about Arabidopsis have shown that the gene SCARECROW (SCR) is expressed in the cortex endodermal initial cells in the root and the stem, regulating the radial organization of the cortex of these organs. The gene is also expressed in the leaf bundle sheath, which is an endodermis. The aim of this paper, based on observations of material from Alismataceae,
Alliaceae, Bromeliaceae, Cyperaceae, Iridaceae,
Rapateaceae, Ruscaceae, Velloziaceae, Xyridaceae and
Zingiberaceae, is to desmonstrate that: a) the procambium gives rise to leaf traces (with protoxylem, metaxylem, protophloem and metaphloem), cauline bundles (with metaxylem and metaphloem only) and pericycle; b) only the pericycle forms the vascular tissues following the procambium activity; c) endodermis with meristematic activity forms all or part of the cortex; d) the plant is an unit: all primary tissues in the root are also present in the stem and in the leaves, and e) there is no
PTM in monocots and thus, in this regard, these plants are not distinct from other angiosperms and gymnosperms.
Evolution of functional diversification in dual phloem systems of cucurbits
Zhang, B 1 ,
Turnbull, C 3
Tolstikov, V 2 , Hicks, L 1 , Fiehn, O 2 ,
2
1 Donald Danforth Plant Science Center, USA;
University of California, Davis, USA; 3 Imperial College
London, UK
By definition, the vascular bundle is a key feature in the evolution of vascular plants. The typical arrangement in angiosperms is abaxial phloem and adaxial xylem in leaves, corresponding to external phloem and internal xylem in stems. However, families such as Cucurbitacaee and Solanaceae deviate from this by developing additional phloem internal to the stem xylem, a configuration known as bicollateral. Cucurbits go further by having extrafascicular phloem (EFP) elements scattered through cortex and peripheral to vascular bundles. The evolutionary advantages of the greater complexity are not yet clear. Cucurbits are popular in phloem studies because of the substantial amounts of sap that can be collected following tissue incision. Yet, it has remained unresolved whether this sap emanates from vascular bundles (fascicular phloem; FP) and/or from the
EFP elements. We have re-examined this question, and also investigated why the sugar content of cucurbit sap is anomalously low. Contrary to expectations, video microscopy reveals that FP very quickly becomes blocked on cutting, whereas the EFP phloem bleeds for extended periods. Thus, previous cucurbit phloem sap studies to date have most likely reported metabolite, protein and RNA composition and transport in the relatively minor extrafascicular sieve tubes. Microdissection and direct sampling of sieve tube contents
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shows that FP does contain ~1 Molar sugars. Moreover, protein profiles in FP differ substantially from those in the extrafascicular sap. Extrafascicular sap is dominated by the well-characterised filament and lectin proteins,
PP1 and PP2, that may function in its slow coagulation.
In contrast, FP contains many major proteins of unknown function, but does include some that may be conserved for wound responses across dicots. The overall compositional differences of the two phloem systems strongly indicate symplasmic isolation and functional divergence. We conclude that the fascicular phloem is largely responsible for sugar transport whereas the extrafascicular phloem may function in signalling, defence and perhaps transport or retrieval of other metabolites. The internal and external components of the bicollateral FP have similar high-sugar metabolite profiles, but may also have divergent functions and/or transport pathways. Possible evolutionary origins of EFP will be discussed in light of the discovery of divergent composition in this family. Ref Zhang et al. (2010).
Evolutionary inferences from transcriptional analysis of wild and domesticated tomato
Sinha, N 1,2
Kimura, S 1
, Maloof, J 1 , Jimenez-Gomez, J
, Dan Chitwood, D
Devisetty, U 1 , Headland, L 1
1 , Fulop, D 1
0 , Koenig, D 0
, Kumar, R 1
,
,
1 University of California, Davis, USA;
Institute, Cologne, Germany;
Tübingen, Germany
3
2 Max Planck
Max Planck Institute,
The domestication of crop species from wild relatives altered human history, while human intervention simultaneously modified the form, physiology, and life history of the domesticated plant species. The process of domestication thus provides us with a outstanding opportunity to understand the process of response to selection. We used genome-wide deep-coverage shortread sequencing to analyze the transcriptomes of one accession of domesticated tomato and three wild relatives for changes in gene expression, coding sequences, and gene regulation. Sampling from seven different organ and tissue types from plants grown in two environments allowed us to capture of the best representation of expressed genes in the tomato complex. We compared our identified transcripts to the recently completed tomato genome sequence and a partially complete sequence of Solanum pennellii, a green-fruited wild relative of tomato, to identify sequence polymorphisms and make differential gene expression calls. In addition, we sequenced miRNA loci, and also placed on the genome repeated regions with a high density of siRNA reads. Our analysis of the transcripts identifies those genes that show the largest expression differences between domesticated tomato and wild species, and in addition allows us to preliminarily identify genes and gene categories that show the strongest signatures of selection.
Caspases and signaling to PCD in self-incompatible
1 pollen
Bosch, M
N (VE) 1
1 , Wilkins, KA 1 , Tudor, RL 1 , Franklin-Tong,
University of Birmingham, UK
Self-incompatibility (SI) is an important mechanism used by many angiosperms to prevent inbreeding. A multiallelic S locus allows discrimination between ‘self’
(incompatible) pollen from ‘non-self’ (compatible) pollen when it lands on the stigma. Papaver rhoeas
(poppy) uses the pistil S determinant (recently renamed as PrsS, Papaver rhoeas stigma S) which is a small novel secreted protein that acts as a signalling ligand that interacts with its cognate pollen S-determinant ( Papaver rhoeas pollen S), PrpS. When PrsS interacts with incompatible pollen, it triggers Ca 2+ influx and transient increases in ROS and NO. Downstream targets/events include: depolymerization of actin and microtubules, phosphorylation of a soluble inorganic pyrophosphatase,
Pr-p26.1, and activation of a MAP kinase, PrMPK9-1.
Ultimately, and probably the major target for SI signals is initiation of programmed cell death (PCD) involving several caspase-like activities in incompatible pollen.
This provides a very neat way to get rid of unwanted
‘self’ pollen and prevent self-fertilization. I will discuss the several caspase-like activities that we have found in poppy pollen. We have evidence that a DEVDase,
VEIDase and a LEVDase are involved in SI-mediated events. As the SI-induced caspase-like proteins, such as
DEVDase, have a very narrow and acidic pH optimum
(~pH5.5), this suggested that SI triggers rapid acidification of the cytosol. Studies have revealed that SI induces rapid changes to the vacuolar compartments, which undergo reorganization and disintegration. This could potentially generate the optimal acidic pH for caspase-like activities. We have recently investigated the role of a Vacuolar Processing Enzyme (VPE) from poppy pollen, as VPEs function in PCD in several plant systems. We found that although it is not required for poppy pollen PCD, it binds to the DEVD tetrapeptide, a signature substrate for caspase-3. Although the recombinant poppy VPE has YVADase activity and requires an acidic pH for activity, our studies revealed several surprising features. I will present a model of how we currently believe these components form a network to mediate PCD in poppy pollen.
Lace plant: a novel model system to study developmental programmed cell death in vivo
Gunawardena, A 1
Wertman, J 1
, Wright, H
, Rantong, G 1
1 , Lord, C 1 , Dauphinee , A 1
1 Dept of Biology, Dalhousie University, Halifax, Canada
Programmed cell death (PCD) is a genetically encoded, active process which results in the death of individual cells, tissues, or whole organs. As in animals, PCD in plants is a normal part of development throughout the life cycle, from death of the embryonic suspensor to
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senescence of leaves and flower parts. Several Monstera species (Araceae) and a single species of the Aponogeton family, Aponogeton madagascariensis (Lace plant), are the only vascular plants that form leaf perforations by
PCD during development. In Lace plant, PCD is initiated in a population of cells at the center of the perforation site, and then proceeds outward through epidermal and mesophyll cells to tissue approximately five cells from the veins. The accessibility and predictability of perforation formation, the ability to perform live cell imaging due to the thin and aquatic nature of leaves, propagation of lace plant in sterile conditions, and the successful regeneration from callus tissue make Lace plant an extremely attractive and tractable system in which to study developmental PCD. In this system, discrete populations of cells undergo changes involving the vacuole, nucleus, chloroplast, mitochondria, actin microfilaments, transvacuolar strands, and cell walls.
Interestingly live cell imaging using whole mounts showed dividing, dumbbell-shaped chloroplasts that persisted until the late stages of PCD and chloroplasts formed a ring around the nucleus. Increased numbers of transvacuolar strands were also observed at this stage and mitochondria formed aggregates and then later moved towards the nucleus. An increasing number of objects, including intact chloroplasts undergoing Brownian motion inside the vacuole, were observed at this stage. A rapid shrinkage of the nucleus and cessation of cytoplasmic streaming, including mitochondrial aggregate movements, likely infer tonoplast rupture.
Indirect evidence using ethylene biosynthesis inhibitors implicated involvement of ethylene in regulating perforation formation. This unique plant is an excellent model for studying developmental PCD in plants.
Cell biological aspect of plant immunity: two vacuolemediated defense strategies
Hara-Nishimura, I 1
Nishimura, M 2
, Hatsugai, N 1
1 Kyoto University, Japan;
Biology, Japan
2
, Tamura, K 1 ,
National Institute for Basic
Because plants do not have immune cells, each cell has to provide its own defense against invading pathogens. A common plant defense strategy involves programmed cell death (PCD) at the infection site, but how the PCDassociated immunity is executed in plants is not fully understood. In this meeting, we provide a novel mechanism underlying cell-autonomous immunity, which involves the fusion of membranes of a largecentral-vacuole with the plasma membrane, resulting in the discharge of vacuolar antibacterial proteins to the outside of the cells where bacteria proliferate (1,2). We found that a defect in proteasome function abolished the membrane fusion associated with both disease resistance and PCD in response to avirulent bacterial strains but not to a virulent strain. Furthermore, RNAi plants with a defective proteasome subunit PBA1 have reduced
DEVDase activity, which is an activity associated with caspase-3, one of the executors of animal apoptosis. The plant counterpart of caspase-3 has not yet been identified.
Our results suggest that PBA1 acts as a plant caspase-3like enzyme. Thus, this novel defense strategy through proteasome-regulating membrane fusion of the vacuolar and plasma membranes provides plants with a mechanism for attacking intercellular bacterial pathogens that invade from stomata of the leaves. The immune system complements another vacuolar defense mechanism in which viral propagation inside the cell is checked by vacuolar collapse (3,4). Our findings suggest that plants have evolved a cell-autonomous immune system based on membrane fusion to inhibit proliferation of bacterial pathogens and a vacuolar-collapse system to be unable to spread systemically viral pathogens. (1)
Hatsugai et al. 2009. (2) Pajerowska-Mukhtar et al. 2009.
(3) Hara-Nishimura et al. 2005. (4) Hatsugai et al. 2004.
Interactions between stress, ROS and senescence in
1 regulating petal cell death
Rogers, H 1 , Mohd-Salleh, F 1 , Foyer, C 2 , Theodoulou, F 3
2
School of Biosciences, Cardiff University, UK;
University of Leeds, UK; 3 Rothamsted Research, UK
Petals and leaves share common evolutionary origins, although they perform very different functions.
Wallflowers ( Erisymum linifolium ), an ornamental species closely related to Arabidopsis , provides a good species in which to study and compare these processes as both petal and leaf senescence can be divided into defined stages. Senescence is associated in many species with an increase in the abundance of ROS and in defence gene expression. Several plant growth regulators (PGRs) have also been implicated in the regulation of petal senescence and cell death, with ethylene as a key regulator in many species including wallflowers. The interactions between PGR- and ROS-signalling during wallflower senescence have been investigated using microarrays, the expression of marker genes, measurement of ROS levels and enzyme activities of enzymes related to ROS production and scavenging.
Recently we have focussed, on the role of SAG21
(AtLEA5), which was identified independently both as a
Senescence Associated Gene (SAG) in Arabidopsis and as an oxidative stress-responsive gene. AtLEA5 encodes a group 3 late embryogenesis abundant (LEA) protein whose precise role during senescence and oxidative signalling is remains to be defined. AtLEA5 is expressed transiently, early in Arabidopsis leaf and wallflower petal senescence. However, the AtLEA5 transcripts are most abundant prior to enhanced H2O2 accumulation. In
Arabidopsis AtLEA5 expression is increased in leaves in response to drought and abscisic acid indicating a general role in defence and stress protection. Moreover, phenotypic analysis of Arabidopsis AtLEA5 overexpressors and antisense lines reveals a clear senescence phenotype. AtLEA5 promoter function is now under investigation with a view to understanding how expression is coordinated by stress and senescence/ cell death signals.
Programmed cell death and plant adaptation to salinity and waterlogging: unravelling mechanisms
1 for ion specific signalling
Shabala, S 1
University of Tasmania, Australia
Programmed Cell Death (PCD) is a fundamental cellular process observed in eukaryotic cells of different origin.
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Being an ordered series of events, PCD facilitates the removal of redundant, misplaced, or damaged cells and is essential for cellular differentiation and tissue homeostasis. PCD also plays an important role in mediating plant adaptive responses to a range of biotic
(pathogens) and abiotic (salinity, cold, waterlogging and hypoxia) stresses. Recent studies suggest that expression of animal anti-apoptotic genes in transgenic plants may significantly improve plant’s ability to tolerate a variety of biotic and abiotic stresses. The underlying cellular mechanisms of this process remain unexplored. In this study, a critical role for ionic homeostasis in triggering
PCD under abiotic stress conditions was studied. Specific ion flux ‘signatures’ were investigated in Nicotiana benthamiana plants transiently expressing animal CED-9 anti-apoptotic gene and undergoing salt- and oxidative stresses [1]. Using a range of electrophysiological techniques, we show that expression of CED-9 increased plant salt and oxidative stress tolerance by altering K + and H + flux patterns across the plasma membrane. Our data shows that PVX/CED-9 plants are capable of preventing stress-induced K+ efflux from mesophyll cells, so maintaining intracellular K + homeostasis. We attribute these effects to the ability of CED-9 to control at least two types of K + -permeable channels; outwardrectifying depolarization-activating K + channels (KOR) and non-selective cation channels (NSCC). A possible model linking CED-9 expression and ionic relations in plant cell was suggested [2] and then validated using
Arabidopsis transport mutants [3]. This model was later extended to explain plant adaptive responses to phytotoxins accumulation in waterlogged soils. Using our experimental data on barley roots [4] we show that
H + -coupled movement of undissociated phytotoxins
(such as phenolic acid) across the plasma membrane results in membrane depolarisation and leads to a significant reduction in the cytosolic K prolonged increase in the net Ca 2+
+ pool. Root exposure to phenolics is also accompanied by a
uptake, most likely through DACC channels. Increases in the cytosolic free
Ca 2+ pool leads to a dramatic rise in ROS levels resulting from [Ca 2+ ]cyt activation of NADPH oxidase via positive feedback mechanisms. This causes an additional K + efflux via ROS-activated NSCC channels, further decreasing cytosolic K + pool and causing the activation of caspase-like proteases that lead to PCD. Thus, it is suggested that the existing ethylene-based models of aerenchyma formation by lysogeny should be complemented by the ethylene-independent signalling pathway: the disturbance to K + homeostasis, as suggested above. The ecological and adaptive significance of above findings is discussed. References: (1) Shabala et al 2007.
(2) Shabala S 2009. (3) Demidchik et al. 2010. (4) Pang et al. 2007. (5) Shabala S 2011.
Developmental biologists and 'housekeeping genes': the whys and wherefores
Tsukaya, H 1
1 Graduate School Science, University of Tokyo, Tokyo,
Japan
184
As mentioned in the abstract of this symposium, an increasing number of ‘housekeeping’ genes have been identified, where mutations cause specific defects in multicellular organogenesis. The housekeeping genes mentioned here include poly(A) polymerase, ribosomal genes, genes for HISTONE MONOUBIQUITINATION1 ,
ELONGATOR ( ELO ) genes, genes for RNA processing and so on. Apparently, these typical housekeeping genes are believed to be required for all cell types. But in these years, such genes were found to be involved in several processes such as an organ-identity-dependent growth regulation, a dorsoventral polarity and meristem differentiation. That was a surprise for me, since when I was a graduate student in the early 1990s, most developmental biologists have mainly focused on transcription factors (TFs) to reveal key mechanisms of cell-type differentiation. Differing from the actual concept where genetic networks controlling organogenesis are spatio-temporally controlled by
ON/OFF switching of specific TFs, the newly emerging viewpoint tells us that the proper expression of
‘housekeeping’ genes is also required for correct spatiotemporal regulations of organogenesis. In this symposium, Dr Michael Lenhard, the co-chair of this symposium, as well as Drs Gorou Horiguchi, Mary
Byrne, Mieke van Lijsebettens, and Munetaka Sugiyama, will introduce to us the latest understandings on the hidden roles of the housekeeping genes on various developmental processes in multicellular organisms. I hope that this symposium will represent a key starting point that should open a new era of studies on organogenesis.
The role of poly(A)-polymerases in controlling identity-dependent growth patterns in organs of
Arabidopsis thaliana
Lenhard, M
JL 4 , Groth, D
1 , Vi, LS
1
2 , de Moor, C 3 , Rao, N 4 , Manley,
1 Institut für Biochemie und Biologie, Universität
Potsdam, Potsdam, Germany; 2 Dept of Cell &
Developmental Biology, John Innes Centre, Norwich,
UK; 3 School of Pharmacy, The University of Nottingham,
Nottingham, UK; 4 Biological Sciences, Columbia
University, New York, USA
Different organs of plants and animals grow to characteristic sizes and shapes that depend on their identity. In fact, the size and shape of an organ are among the most important criteria we use when determining its identity, for example as a sepal or a leaf.
Work over the last 20 years has revealed in molecular detail how the identity of an organ is controlled; however, we still know surprisingly little about how organ identity modified patterns of growth within the primordium to achieve the correct final size and shape.
Here, we report on a novel mutant in Arabidopsis thaliana that forms smaller leaves, but bigger flowers.
Intriguingly, the effect of the mutation depends on the identity of the organ, not its position on the plant, marking out this mutation as a promising entry point to studying the relation between organ identity and growth.
The affected gene encodes one of three nuclear poly(A)polymerases in Arabidopsis involved in polyadenylation of pre-mRNAs. Using an mRNA fractionation approach,

we have defined a set of mRNAs whose poly(A)-tails and stability are changed in the mutant compared to wild type, suggesting functional specialization among the different poly(A)-polymerase isoforms in plants.
Ongoing work on understanding the connection between this poly(A)-polymerase and identity-dependent growth patterns in plant organs will be presented.
General and specific roles of the ribosome in leaf development
Horiguchi, G 1 , Tsukaya, H 2
1 Dept of Life Science, College of Science, Rikkyo
2 University, Japan; Graduate School of Science, The
University of Tokyo, Japan
The ribosome is essential in life as translational apparatus while it also regulates expression of specific genes. In
Arabidopsis thaliana , all ribosomal proteins (r-proteins) are encoded by small gene families. Thus, viable rprotein defective mutants have been available. Mutations in r-proteins perturb cell proliferation, cell expansion, vasculature development and leaf morphogenesis.
Whether these phenotypes are caused by overall ribosome insufficiency or partial dysfunction of the ribosome caused by the deficiency of a particular ribosomal protein is not known. To distinguish these possibilities, a comparative study using a sufficient number of r-protein mutants is required. We have isolated a number of r-protein defective mutants. Most of these mutations were associated with pointed leaves having reduced growth and cell proliferation activity. In addition, leaf abaxialisation was usually observed when these r-protein mutations were combined with asymmetric leaves1 (as1) or as2. Individual r-protein mutants exhibit variable phenotypic strengths. When the number of leaf palisade cells and degree of enhancement for as2 leaf polarity defects were compared across various r-protein mutants, there was a correlation between the strengths of these two phenotypes. This result suggests that ribosome insufficiency is a general causer of these phenotypes and phenotypic variations can be attributable to the degree of functional redundancy among paralogues of a given r-protein family. However, several r-protein mutants showed a preference toward a specific developmental defect. rpl4d mutations did not affect cell proliferation but enhanced expansion of leaf palisade cells. Moreover it caused strong abaxialisation of leaves in the as1 and as2 backgrounds. On the other hand, rps28b enhanced leaf abaxialization of as2 at a weaker extent expected from its negative effect on cell proliferation. In addition, hypomorphic rps6a alleles exerted the strongest effects on most of the phenotypes examined. These findings suggest that deficiencies of these three r-protein genes lead to the production of dysfunctional ribosomes. It might be possible that depending on their structural abnormalities, dysfunctional ribosomes affect translation of specific transcripts involved in the regulation of some leaf developmental processes.
Genetic interactions in the plant shoot apex: a role for the ribosome in meristem function and leaf development
Byrne, M 1
1 University of Sydney, Australia
Plant shoots are characterized by the presence of a shoot apical meristem, which is established in the embryo and maintained throughout growth of the plant. The shoot meristem iteratively produces lateral organs and ultimately gives rise to all organs of the plant shoot. The meristem also influences the final shape or pattern of a leaf. Early in development leaves establish dorsoventral polarity. Setting up leaf dorsoventral polarity depends on signalling from the meristem to the initiating leaf, as well as concerted interactions between dorsal and ventral sides of the leaf. Control of meristem function and leaf patterning occurs via a network of gene interactions and involves transcription and post-transcriptional mechanisms. Using Arabidopsis thaliana as a model system to study plant development we have identified new players in the gene network controlling plant shoot development as ribosomal proteins. Mutations in ribosomal proteins have specific developmental defects in meristem function and leaf patterning in the embryo and in the shoot. We propose that the ribosome has more than a housekeeping function, and is involved in regulating growth and patterning in plant development.
The Elongator complex regulates specific auxin-
1 related genes at RNAPII transcription elongation
De Groeve, S
Tsukaya, H 3
A 6 ,
1 , Nelissen, H 1 , Fleury, D
, Van Der Straeten, D
Van Lijsebettens, M 1
VIB, Ghent University, Belgium; 2
3
4
2 , De Jaeger, G
, Bitonti, B 5
1 ,
, Houben,
ACPFG, University of
4
Adelaide, Australia; NIBB/ University of Tokyo, Japan;
Ghent University, Belgium; 5 University of Calabria,
Italy; 6 IPK, Gatersleben, Germany
Growth is a complex trait that is steered by developmental, physiological and biochemical pathways.
We study the genetic and epigenetic control of organ growth in plants by using a mutational approach in
Arabidopsis and leaves as the experimental system. Leaf growth occurs mainly in two dimensions and final leaf size and shape is determined by cell number and cell volume. Over the last decades, a number of transcription factors have been identified that control leaf initiation and early stages in patterning and growth. The next step in the research is to identify and study the upstream mechanisms that regulate the transcriptional activities.
Histone modification during transcription is one of these upstream regulatory mechanisms since our leaf growth mutants identified conserved chromatin modifying complexes, HISTONE MONOUBIQUITINATION1
(HUB1) and ELONGATOR with a role in RNApolII transcription activation and both affecting organ formation by regulating cell number (Nelissen et al.
2005; Fleury et al. 2007). Elongator contains a GCN5type histone acetyl transferase that targets specific auxinrelated genes during transcription elongation as demonstrated by transcriptome analysis followed by chromatin immune precipitation. Typical auxin-related phenotypes in venation patterning and plant architecture, and auxin sensitivity in the Elongator mutants showed the biological relevance of the transcriptional regulation by Elongator (Nelissen et al. 2010). We postulate that the intrinsic activity or allelic strength of histone modifying complexes might contribute to the leaf size and shape
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and plant architecture in species and that their putative cross-talk to the environment might regulate plasticity in growth.
Differential requirement for the supply of essential
RNAs, such as snRNAs and rRNAs, in various aspects of plant development
Sugiyama, M 1 , Ohbayashi, I 1 , Ohtani, M 2
1 Botanical Gardens, Graduate School of Science, The
University of Tokyo, Japan; 2 Biomass Engineering
Program, RIKEN, Japan
In an attempt of genetically identifying hidden machineries of plant development, we isolated many temperature-sensitive mutants of Arabidopsis and characterized them with a predominant focus on organogenesis in vitro. Two of these mutants, srd2 and rid1, exhibit severe temperature sensitivity for hypocotyl dedifferentiation (acquisition of cell proliferation competence) and de novo formation of meristems but are not much affected in callus growth and activation of preexisting meristems. Another mutant, rid2, is also highly temperature-sensitive for hypocotyl dedifferentiation and meristem neo-formation but displays a little broader spectrum of phenotypes than srd2 and rid1. The responsible genes of these mutants, SRD2, RID1, and
RID2, have been shown to encode an snRNA transcription activator, a putative RNA helicase implicated in pre-mRNA splicing, and a nucleolar methyltransferase-like protein involved in rRNA biosynthesis, respectively. Detailed analyses of these genes/mutants have indicated that snRNAs and rRNAs can be limiting factors of the competence for cell proliferation, probably through the restriction of the capacities of pre-mRNA splicing and translation, that their supply is differentially required for various aspects of development, and that their supply level is dynamically controlled during development. Recently, we have found that sriw1, a mutation in the gene encoding a NAC-family transcription factor ANAC082, alleviates the defects of all the three mutants. This finding raises the possibility that the supply level of essential RNAs is monitored to tune cell proliferation by a mechanism involving ANAC082.
Origin, phylogeny, adaptive radiation, and geographic diversification of Bromeliaceae
Givnish, T 1
K 5
, Barfuss, M 2
, Horres, R 6
, Van Ee, B 3
, Gonsiska, P
, Riina, R
1 , Jabaily, R 7
4 , Schulte,
, Crayn, D 5 ,
Smith, A 8
Zizka, G 1
, Winter, K
, Berry, P 1
9 , Holst, B 10
, Arbor, A 1
, Luther, H 1
, Sytsma, K 1
, Till, W 1 ,
1
2
6
University of Wisconsin–Madison, Madison,USA;
University of Vienna, Austria;
University, Spearfish, USA;
Arbor, USA; 5
University, UK; 9
4
3 Black Hills State
University of Michigan, Ann
James Cook University, Cairns, Australia;
GenXPro, Frankfurt Am Main, Germany;
Dominion University,Virginia Beach, USA;
7 Old
Smithsonian Tropical Research
Institute, Balboa, Panamá;
Gardens, USA
8 Oxford
10 Marie Selby Tropical
Sequence variation in eight rapidly evolving plastid regions for 90 bromeliad species from 45 of 56 described genera confirm that the eight bromeliad subfamilies are related to each other in ladder-like fashion:
(Brocchinoidieae, (Lindmanioideae, (Tillandsioideae,
(Hechtioideae, (Navioideae, (Pitcairnioideae, (Puyoideae,
Bromelioideae). Puya shows a basal split between species found primarily at higher elevations in the Andes and those found near sea level in Chile. The earliestdivergent bromelioids are mostly restricted to the southern Andes and Chilean coast. We calibrated this phylogeny against the ages of fossil monocots using penalized likelihood, and assessed patterns of biogeographic spread using maximum parsimony,
Bayesian inference, and S-DIVA. Bromeliads appear to have arisen in the Guayana Shield roughly 100 Mya, spread centrifugally in the New World, and reached tropical West Africa via long-distance dispersal 9.2 Mya.
Modern lineages began to diverge from each other 19
Mya, with invasions of drier or higher peripheral areas in
Central America and northern South America beginning
16–13 Mya, coincident with a major adaptive radiation – the 'bromeliad revolution' – involving the repeated evolution of epiphytism, the tank habit, CAM photosynthesis, and avian pollination, as well as several features of leaf and trichome anatomy, and an accelerated pace of species diversification. This revolution coincided with the uplift of the northern Andes and its invasion by epiphytic tillandsioids and by ancestors of the pitcairnioids, puyoids, and bromelioids. Bromelioids invaded the Serra do Mar and nearby mountains in southeastern Brazil, most likely from southern Chile, starting 9.1 Mya. A major radiation of epiphytic bromelioids in Brazil began 5.7 Mya. Calculations of net rates of species diversification for subfamilies or clades of similar rank range from 0.16 sp sp -1 My -1 in brocchinioids to 1.12 sp sp -1 My -1 in the tank-epiphytic bromelioids. Acquisition of the epiphytic habit and related traits appears to have accelerated net species diversification by favoring seed traits that increased attachment to epiphytic perches, and coincidentally increased the ability of tillandsioids and bromelioids to colonize extensive montane regions in the Andes and
Central America, and in the Serra do Mar and nearby mountains in coastal Brazil, permitting geographic speciation to proceed in massively parallel fashion as epiphytes occupied a cloud-forest landscape dissected by numerous drier, lower valleys that could act as extrinsic barriers to gene flow. Avian pollination (mainly by hummingbirds) evolved at least twice, coincident with the invasion of cool, wet montane habitats. Net diversification rates were significantly higher in hummingbird-pollinated clades, perhaps reflecting the rise of gullet-shaped flowers adapted to such pollinators
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and the resulting opportunity for rapid partition¬ing of another rapidly speciating montane clade based on differences in bill length. Entomophily is the ancestral condition in bromeliads, and bat pollination appears to have evolved several times from hummingbird pollination; chiropterophily is associated with warmer and/or drier conditions then ornithophily.
Inferences for the biogeography of Bromeliaceae based on Protoananaceae, a new monocot fossil family from Early Cretaceous of northeast Brazil
Leme, EMC 1 , Brown, GK 2
1 Herbarium Bradeanum, Brazil;
Wyoming,USA
2 University of
Frequently, new fossil materials are discovered that suggest earlier diversifications than had been estimated by molecular data for extant taxa. That appears to be the case of Protoananas lucenae , the single species of the new fossil angiosperm family Protoananaceae, which represents an extinct putative lineage of monocots from the Early Cretaceous, Crato Formation limestone (ca. 114 to 112 Mya) in the Araripe Basin, northeastern Brazil.
The peculiar character combination of Protoananas lucenae , composed by relatively advanced structures, brings to light new interpretative possibilities for the biogeography of its putative closer relative, the extant
Bromeliaceae, and suggests that possibly other primitive lineages of bromeliophytes predate or co-existed with this fossilized taxon. Some important features of
Protoananas lucenae are the strobilate, ellipsoidal inflorescence with conspicuous suborbicular bracts surrounding its base, the apparent epigynous flowers, abundance of a resin-like substance in scape, scape bract, and floral tissues, the presence of objects similar to epicuticular wax that resemble the Strelitzia-type, and the entire leaf-like scape bract with pronounced parallel nerves slightly convergent toward the apex, bearing stomata apparently of the paracytic type disposed in rolls along the intercostal zone. However, the most intriguing structure consists of fragments interpreted as peltate trichomes which are compatible, in terms of topology, architecture and structure, with the peltate trichomes found in extant Bromeliaceae. Since the absorbing function of the peltate trichome in extant taxa can be interpreted as an adaptation for low soil fertility, or for habitats in which root absorption is not effective, like some periodically or permanently flooded areas in the present Brazilian swampy Atlantic Forest, the presence of these structures in Protoananas lucenae is compatible with the generally conceived paleoenvironment of the
Araripe Basin and so the putative paleohabitat of this taxon associated with comparatively humid, restrict sites along marginal lacustrine or riverside environments, despite the general warm and arid or semiarid paleoclimate of this period. The late Aptian age of
Protoananas lucenae , significantly older than the most recent hypothesized diversification estimated by molecular data for its putatively closest relative, preceding the final separation of Africa and South
America around 110-90 Mya, allows the reasonable speculation that Protoananas, and related lineages, had ample time to disperse throughout Gondwana. The seemingly advanced morphological features of
Protoananas lucenae , as the putative ancestral stemlineage of Bromeliaceae, may help to explain the origin of the family and its actual geographical range, suggesting the Brazilian shield as the hypothetically center of origin of the group, with subsequent radiation and diversification throughout South, Central and North
America, and Africa. In addition, the progressive population isolation and high extinction rates caused by the desertification during Tertiary and Quaternary in
Africa may help to explain the current distribution of
Bromeliaceae in the African continent, possibly together with isolated episodes of recent long-distance dispersal.
Bromeliaceae diversity in Brazil: current status and research perspectives
Wanderley, MGL 1
LM 3 , Lousada, RB 0
, Benko-Iseppon, AM
, Sousa, GM 5
2 , Versieux,
1 Instituto de Botânica, São Paulo, Brazil;
Federal de Pernambuco, Recife, Brazil;
2 Universidade
3 Universidade
5
4
Federal do Rio Grande do Norte, Natal, Brazil;
Universidade de São Paulo, São Paulo, Brazil;
Universidade Federal do Piauí, Teresina, Brazil
It has long been accepted that Brazil, particularly the forests along the Atlantic coast are among the most important centers of diversity of Bromeliaceae.
According to a recent official Brazilian checklist the country harbors 42 genera and 1,207 species of bromeliads from which 22 genera are endemic. Improved knowledge of Brazilian Bromeliaceae flora allowed new assessments about species delimitation, distribution, conservation and evolution. The present work brings an insight regarding traditional systematic revisions of
Brazilian endemic groups, as well as new approaches employing molecular techniques, in order to understand the radiation of these plants. The basis of this review regards important herbarium collections kept in different institutes and universities, with emphasis on the
Herbarium SP in the Instituto de Botânica (IBt) in São
Paulo that has a long tradition in the research of bromeliads since its foundation in 1938. The Herbarium
SP is very representative for the family and has been used as base for the development of several distinct taxonomic investigations. Moreover, this institution acts as a traditional center to form new Bromeliaceae specialists conducting monographs of distinct genera
(e.g. Aechmea subg. Chevaliera , Alcantarea ,
Cryptanthus , Encholirium , Orthophytum , Portea ,
Tillandsia subg. Anoplophytum , and Vriesea ). Besides the traditional morphological revisions, new approaches are being used, sometimes in partnership with colleagues from abroad. Recent projects include molecular phylogenetics of distinct groups (e.g. Alcantarea and
Orthophytum ) in collaboration with the Universidade
Federal de Pernambuco (UFPE), Universidade Federal do Rio Grande do Norte (UFRN), Johann Wolfgang
Goethe University (Frankfurt, Germany) and Royal
Botanic Gardens, Kew (UK). Pollen data, cytology, as well as leaf anatomy are additional tools that we have employed to characterize Bromeliaceae taxa. A molecular biology laboratory is now available at IBt, being active in projects regarding population genetics and phylogeographic patterns in Bromeliaceae that occupy areas of high endemism with emphasis on species from the Espinhaço and Mantiqueira mountain ranges and the Atlantic rain forest as whole. In the UFPE
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research lines are focused on the phylogeographic, cytology and population genetics of several taxa, with emphasis on the genera Cryptanthus , Dyckia and
Encholirium . The establishment of an intensive program of collection in the wild and cultivation in green houses has allowed us to create an expressive living collection.
The investigation of the evolutionary relationships and taxonomic delimitation within closely related genera and species complex are now the challenge to a better comprehension of the family in Brazil. Considering this scenario, the next steps include efforts to clarify the limits among these problematic taxa. Additionally, we maintain our research regarding floristic inventories in different areas, ranging from whole states (e.g. São
Paulo, Piauí, Minas Gerais, Rio Grande do Norte, Ceará, and Pernambuco) to national and state parks (e.g. Parque
Nacional da Serra do Cipó, Parque Nacional da Chapada
Diamantina, and Parque Estadual do Rio Preto, among others) documenting the diversity and describing new taxa. As challenges for the near future, we recognize large groups demanding detailed revisions and some interdisciplinary researches including molecular, cytological, palynological, ecological, and anatomical evaluations.
Molecular phylogenetics of the Brazilian giant
Bromeliads (
Alcantarea
, Bromeliaceae): implications for morphological evolution and biogeography
Versieux, L
1
Lexer, C 2
1 , Barbará, T 2 , Wanderley, MDG 3 , Fay, M 4 ,
Universidade Federal do Rio Grande do Norte, Natal,
3
Brazil; 2 University of Fribourg, Fribourg, Switzerland;
Instituto de Botânica, São Paulo, Brazil; 4 Royal Botanic
Gardens, Kew, Richmond, UK
The genus Alcantarea comprises 26 species endemic to rocky outcrops from eastern Brazil. Most species are ornamental and several are threatened due to habitat loss and over collection. In this paper we examine the phylogenetics of Alcantarea and its relationship with the genus Vriesea , of which Alcantarea has been treated as a subgenus. We discuss the morphological evolution of the stamen position and its implication for pollination and the occurrence of Alcantarea in campo rupestre vegetation. Hybridization hypotheses associated with the genus are discussed.DNA sequence data derived from two plastid markers ( trn Krps 16, trn Cpet N) and from a low copy nuclear gene (Floricaula/Leafy) were combined with 20 nuclear microsatellite loci to construct phylogenetic trees for the genus. Alcantarea is well supported as monophyletic in both Bayesian and parsimony analyses, but sections of Vriesea appear paraphyletic. Microsatellites delimit geographically isolated species groups. Nevertheless individuals belonging to a single species may be closely related to species groups that cluster with other species, suggesting that hybridization may be more frequent then previously thought. Alcantarea brasiliana is identified as putative hybrid between A. imperialis and A. geniculata .
Chiropterophily evolved multiple times within the genus, and invasion of campo rupestre vegetation by Atlantic rainforest ancestors occurred multiple times as well.
Phylogeny and evolutionary history of Pitcairnioideae s. str. (Bromeliaceae) inferred from chloroplast and nuclear data
Weising, K 1 , Schütz, N
S 1 , Zizka, G 2
Schulte, K 4
1 , Wagner, N
, Michalak, I 2
1 , Krapp, F 1
, Silvestro, D 2
, Blank,
, Ibisch, P 3 ,
2
1 Institute of Biology, University of Kassel, Germany;
Botany and Molecular Evolution, Research Institute
Senckenberg and Goethe-University, Frankfurt,
Germany; 3 Faculty of Forest and Environment,
4
University of Applied Sciences, Eberswalde, Germany;
Australian Tropical Herbarium, James Cook University,
Cairns, Australia
According to the revised infrafamilial classification of
Bromeliaceae by Givnish et al. (2007), subfamily
Pitcairnioideae s.str. includes five genera: Pitcairnia
(including Pepinia ), Fosterella , Deuterocohnia
(including Abromeitiella ), Dyckia and Encholirium . We analyzed phylogenetic relationships within the subfamily, based on sequence data at three (or more) chloroplast
DNA (cpDNA) loci and the nuclear PHYC gene. We also applied AFLPs to achieve better resolution at the level of closely related species groups. The large genus Pitcairnia
(~380 species distributed from Central America to
Argentina) is paraphyletic in the cpDNA tree and takes a basal position in the subfamily. Fosterella (31 species) is monophyletic and sister to a well-supported clade consisting of Deuterocohnia (18 species), Dyckia (~140 species) and Encholirium (28 species). Within Fosterella , six well-supported evolutionary lineages could be defined based on a 6-locus cpDNA tree. Application of a molecular clock and ancient area reconstruction suggest that Fosterella originated in the mid Miocene about 10 mya, in an area now known as the Yungas at the eastern slopes of the Bolivian Andes. The Andean genus
Deuterocohnia is clearly paraphyletic in the cpDNA tree but not in the nuclear tree, suggesting an ancient chloroplast capture event. A separate status of
Abromeitiella receives no support from the molecular data. The numerous Dyckia species are distributed in azonal habitats of Neotropic savannas and the Atlantic forests of Brazil and adjacent countries, whereas
Encholirium is confined to eastern Brazil. Sequence variation within Dyckia turned out to be extremely low, and phylogenetic trees are hence poorly resolved.
Nevertheless, several clades could be identified that roughly correspond to the geographic origin of the samples. Whereas Dyckia is apparently monophyletic, the status of Encholirium is still ambiguous due to insufficient sampling. The dated chloroplast phylogeny provides evidence that Dyckia experienced a very recent radiation, probably starting less than 1 Mya. Somewhat surprisingly, the earliest branching lineages of Dyckia were collected in eastern Brazil, in the area of codistribution with Encholirium . This would suggest that colonization of the present distribution area of Dyckia occurred in northeast–southwest direction. Our current research aims at – in cooperation with Brazilian partners
– identifying possible adaptations associated with that spread and the rapid diversification that followed.
188

Systematics, evolution, and phylogeography of
Tillandsia
(Bromeliaceae) and related genera
Barfuss, MHJ 1 , Till, W 1 , Samuel, R 1
1 Dept of Systematic and Evolutionary Botany, University of Vienna, Austria
Tillandsia L. is the largest genus within Bromeliaceae comprising more than 660 accepted species in six recognized subgenera ( Tillandsia subgen. Allardtia , T.
subgen. Anoplophytum , T. subgen. Diaphoranthema , T. subgen. Phytarrhiza , T. subgen. Pseudalcantarea , T. subgen. Tillandsia ). Eight additional genera have been associated with Tillandsia in subfamily Tillandsioideae, i.e., Alcantarea , Catopsis , Glomeropitcairnia , Guzmania ,
Mezobromelia , Racinaea , Vriesea , and Werauhia . The segregation of a group of Tillandsia species occurring in
Mexico (Viridantha) and the transfer of xerophytic
Vriesea species to Tillandsia is not accepted by all bromeliad researchers based on long-established morphological characterizations of traditionally accepted genera. Habit shifts from phytotelms to extreme xerophytes, life form transitions from terrestrials to epiphytes and lithophytes, shifts in pollination syndromes, and seed and stigma morphology are key events and characters for the evolution and systematics of genera and infrageneric units, but they can only be interpreted in the context of DNA sequence data, since some of these features have evolved independently within different phylogenetic lineages. Therefore we conducted DNA sequence analyses from nuclear genes
(i.e., PRK and PHYC, totaling about 2500 bp) and published chloroplast markers (with about 6500 bp) in combination with a reevaluation of morphological characters. Based on these results three new genera with a distinctive stigma and seed morphology are segregated from Tillandsia , i.e., Josemania , Lemeltonia , and
Rothowia . Species of all three belonged to the former subgenus Phytarrhiza , which turned out to be highly polyphyletic. Circumscription of the remaining
Tillandsia subgenera also changes significantly and new infrageneric taxa and groups emerge (e.g., T.
subgen.
Pseudovriesea , T.
subgen. Viridantha ). DNA data suggest that the subfamily had its origin in the geologically old parts of northern South America. From there the two earliest diverging lineages migrated into the
Caribbean ( Catopsis , Glomeropitcairnia ) with some taxa extending also into the Andes and into Central America
( Catopsis ). The next diverging clade (Vrieseeae) splits into two subgroups, one radiating into eastern Brazil
( Alcantarea , Vriesea ), the other spreading into the Andes
( Cipuropsis , Werauhia ). Within Tillandsieae the earliest lineage is Guzmania with a predominately Andean distribution, but also extending into Central America and the Caribbean. Early diverging taxa of the core
Tillandsieae have their current distribution mainly in the
Andes of northern Peru, Ecuador and Colombia, which seems to be the ancient area for the whole Tillandsia s.l. complex. Tillandsia s.str. has two centers of diversity, one in the Northern and Central Andes, the other in the mountain systems of Northern Central America, which proves to have reached Mexico and adjacent areas during at least three independent colonization events from the
South ( T. subgen. Pseudalcantarea , T. subgen. Tillandsia p.p., and T.
subgen. Viridantha p.p.).
An overview of the renovation of the Paris
2
Herbarium
Poncy, O 1 , Labat, JN 2 , Dennetière, B 2
, Chagnoux, S 2
, Pignal, M 2 ,
1
Chesselet, P 2
Muséum National d'Histoire Naturelle, Paris, France;
MNHN, Paris, France
Facing for years with saturation, inadequate conditions for storaging and managing the botanical collections, the
Paris Herbarium needed a profound renovation. This ambitious project was launched in 2006 by the Museum national d’Histoire naturelle. It aims to reach three major objectives: increasing the collections storage capacity within the existing building; creating a controlled environment for collection preservation; and renovating the collection itself. The renovation of the building is being implemented in successive phases concerning both managing areas and collection storage space. The work started in August 2009 and should be completed by the end of 2012. Eight collection rooms 2400 m 2 fitted with compactors will allow doubling the storage capacity. The two first renovated rooms were ready in September 2010 and dedicated to cryptogams. T-RH will be monitored but not controlled at low temperature, so researchers will be able to work inside the collection rooms; this will be mitigated by strong IPM. The renovation of the collection, which is estimated to comprise about 10 million specimens, includes three operations: (1) The decades-old backlog (more than 10% of the total) is currently being mounted and sorted for final integration in the general collection. Treating the backlog has started in 2008, it takes place in the building itself and hires 20 people over 3 years. (2) A virtual herbarium is being created: scanning of all herbarium sheets needs an industrial process able to produce up to 10 000 images a day. Images are indexed in the P database (Sonnerat) and available on the MNHN website (www.mnhn.fr) (3) The arrangement of the phanerogam collections is being modified in two ways. Up to now it is split into main geographic areas on the 3 galleries, and the specimens in each of them arranged after Durand’s sequence. They are currently being reorganized in a unique series, and this series is updated to follow the new APG3 linear sequence of Angiosperm families. Reconditioning, scanning and reorganizing is processed by a private company employing 40 people over 2 years. It takes place in a large warehouse like space outside the Museum. It follows the phases of the renovation of the building. The families are being treated successively in the order of the new arrangement. The collections are being reinstalled in the compactors as soon as each room is renovated. Once the whole renovation project will be completed, the Paris
Herbarium (P and PC) will provide optimal facilities to serve the major objectives of biodiversity studies and easy access to the entirety of its extremely rich holdings.
The Paris virtual herbarium will be central for outreach and citizen science; using Web 2.0 approach, the virtual herbarium should be turned into a proper database.
189

Historical botanical collections of G.B. Hinton (1882–
1943)
Rico Arce, MDL 1 , Moreno Gutierrez, E 2
1 Herbarium, Library, Art & Archives, Royal Botanic
Gardens Kew, UK; 2 Comisión Nacional para el
Conocimiento y Uso de la Biodiversidad, Mexico
G.B. Hinton was born in London (UK) in 1882, one of four sons of C.H. Hinton, and Mary Everest Boole both writers and mathematiscians. G.B. Hinton was also nephew of George Everest, whose name was given to the world`s highest mountain. In 1911 G.B. Hinton went and worked in Mexico for various mining companies. He held many positions but by training was an ore-testing specialist. He was an amateur botanist of note and collected many specimens for the Royal Botanic
Gardens, Kew amongst other 12 herbaria around the world. The years between 1936–1940 were the most productive of his life. He died in Mexico in 1943.
Databases of his collections at Kew have been produced for 8 families and have been georeferenced by
CONABIO, covering 68000 sq. km of Mexico’s mountain areas, many of which are now occupied by towns and small cities. He discovered six genera and 360 species new to science. Many types have not been found/collected again. Vegetation relicts are found in the area but these are inaccessible. The paper includes major events of his botanical life, information extracted from the Kew archives and map of the localities and where the endemic of key taxa are/were found with the aim of carrying out a possible plant rescue.
Collections, types, names, databases and digital images: the story of an historically significant liverwort collection in modern times.
Price, MJ 1 , Hinden, H 1
1 Conservatory and Botanical Garden of Geneva (G),
Switzerland
The bryologist Franz Stephani left behind him a mixed legacy: a geographically and taxonomically diverse collection of liverworts, his notorious 'opus magnum'
Species Hepaticarum , 11 000 illustrations of liverworts from his collection ( Icones Hepaticarum ), over 5000 new liverwort names and a trail of nomenclatural disasters of varying sizes. The Conservatory and Botanical Garden of
Geneva (G) houses a large collection of liverworts that is rich in types and includes the Franz Stephani collection.
The institution is also home to the long-running nomenclature based project Index Hepaticarum , for liverwort names published between the effective dates of
1753 and 1973. These two entities, the liverwort type collection and Index Hepaticarum , have been brought closer together through the digitizing of the type specimens in G via the GBIF.ch and the Andrew W.
Mellon Foundation Global Plants Initiative. The historical liverwort collection in G, the nomenclatural database Index Hepaticarum , Stephani’s Icones
Hepaticarum illustrations and the digitized types can now be linked in a virtual environment. Providing electronic access to information on names, types and illustrations in the Stephani collections will hopefully help to surmount some of the more negative aspects of
Stephani’s liverwort legacy.
Historical collections for Flora of Nepal: appeal of
National Herbarium for repatriation
Shrestha, KK 1 , Watson, MF 2
2
1 Central Dept of Botany, Tribhuvan University, Nepal;
Royal Botanic Garden, Edinburgh, UK
The first record of plant collections for Flora of Nepal dates back to early 19th century, when Francis
Buchanan-Buchanan from the United Kingdom, collected plant specimens from the southern part of Nepal–India border to Kathmandu Valley during 1802–1803. He had collected more than 500 specimens from Nepal, and majority of the specimens were reported as new to science. His entire collections were deposited in the
Herbarium of the Linnean Society of London (LINN) and
The Natural History Museum London (BM). Later, another expert from the United Kingdom, Dr Nathaniel
Wallich, sent his colleague Edward Gardner to Nepal in
1818, followed by his personal visit to Nepal during
1820–1821. Their visits extended collection of specimens from Kathmandu to northern part of Nepal-China border, resulting in to collection of more than 2,000 specimens.
They collected several duplicate specimens, and the collections were sent to various herbaria of the world (K-
W, BM, RBGE, CAL, G-DC, etc.). Similarly, J.D.
Hooker visited far-east Nepal in 1848, and J.F. Duthie visited far-west Nepal during 1884–186. Several new species were described based on their collections; however, none of these specimens, even the duplicate specimens, are deposited in the National Herbarium of
Nepal (KATH), the country of origin. It has been urgently felt that the National Herbarium should appeal for the repatriation of a set of such historical duplicate specimens, especially from the British Herbaria (LINN,
BM and K-W). Attempts should also be made to explore the occurrence of these historical species in the natural habitats, and prepare new sets of herbarium specimens, wherever possible. However, due to lack of proper field data and poor herbarium labels, it is being extremely difficult to trace the localities of such historical collections. Alternative approach to restore the historic collections in the country of origin would be the repatriation of historical collections in the form of a
‘Virtual Herbarium’, with high quality digital images of the specimens. Based on the field data or herbarium label, extensive botanical explorations should be organized jointly for the re-collections of the specimens from the place of their origin. Such collaborative effort would be instrumental in recording the specimens, assessing and monitoring of their occurrence, before their extinction from nature. A collaborative project between the British herbaria (BM, E, K) and Nepal herbaria
(KATH and TUCH) would be ideal to develop a Virtual
Herbarium of the historical collections, as well as to organize botanical explorations for documenting the historical specimens from the natural habitats.
1
The herbarium as time capsule
Flannery, M 1
St John's University, NY, USA
190

The importance of herbaria in systematics is obvious, as is their function in studying environmental change. Here
I would like to discuss another use for herbaria, that is, as historical documents with which to study the society and culture within which the collections were created. My case study deals with the herbarium at the Museum of
Natural History in Providence, Rhode Island in the US. It contains about 6,000 specimens which makes it a small one, but to me it is interesting because almost all these specimens were collected during the late 19th and early
20th century. It is a collection which is not presently being added to, and this is why the Museum’s curator,
Dr. Marilyn Massaro, refers to it as a 'time capsule,' a selection of pressed plants that were stored away and serve as a window into the collecting done at a certain time by people living in a particular area. The Natural
History Museum in Providence was created in 1896 by a group of wealthy patrons, many of whom had their own natural history collections which they contributed to the
Museum. These items serve as the basis for its collections, which, besides herbarium sheets, include minerals, animal remains, and anthropological items as well. While the museum has up-to-date exhibits in anthropology, biology, and astronomy (its official title is the Natural History Museum and Planetarium), it has a feel similar to what it must have had when it first opened.
There have been few alterations to its architecture which is a combination of French chateaux style with American
Arts and Crafts. The collection mirrors this reference to the past, and I would argue that there is value in its very lack of attention to present-day collecting practices. Yes, there are many other collections that have sheets as old as those found here, and many, of course, with sheets that are much older and more valuable. But the latter are often stored in among sheets from many other eras. This is the value of a time capsule: its items are all of one era and thus give a glimpse of that era. Studying this herbarium reveals what the collectors valued and where they went. Some of the sheets are of local flora, and
Rhode Island having a great deal of shoreline, seaweeds and grasses are found in abundance, but there are also plants from areas where the Museum’s patrons vacationed. This collection could be useful to social historians studying the class structure, to cultural historians looking at the pastimes, to civic historians investigating how the Museum’s past, and even to historians of religion since natural theology was a driving force for 19th-century collecting. While I am not in any way denigrating the botanical importance of this herbarium, I am emphasizing that it could be a resource for historians as well. And since there are other herbaria like this, its use in historical research could serve as a model for investigations at other institutions.
Valuing a change in the management of Australian biological collections
Mwebaze, P 1 , Bennett, J 1
1 Crawford School of Economics and Government, The
Australian National University, Canberra, Australia
Biological collections maybe underutilised because of transaction costs incurred in accessing them. The benefits generated by collections are often undervalued by policymakers. One of the ways to reduce transaction costs and foster greater utilisation of biological collections that could generate larger net benefits to society is through the creation of a central database of biological collections, available online. This paper develops a framework for the valuation of the impacts of a policy option associated with the provision of biological collections. To measure the benefits of the policy change, a Contingent Valuation (CV) survey was conducted for the primary users of biological collections.
Marginal willingness to pay (WTP) to purchase access to the new central database was investigated through a payment vehicle of an annual subscription user fee.
Approximately 60% of institutions currently using
Australian collections are willing to contribute funds to maintain the database in the long-term. Results reveal a positive mean WTP of between AUD230 and AUD356 per user per year, depending on the model used.
Expenditure figures support the CVM estimates obtained.
These findings may be useful for resource allocation decisions regarding Australian biological collections.
Angiosperms from the Cretaceous (Campanian)
Barykov flora of the Ugol’Naya Bay (north-eastern
Russia)
Moiseeva, M 1 , Herman, A 1 , Spicer, R 2
1 Geological Institute, Russian Academy of Sciences,
Moscow, Russia;
UK
2 The Open University, Milton Keynes,
The Barykov Flora from the Upper (Coaly) Member of the Barykov Formation in the Ugol’naya Bay (North-
Eastern Russia) is dated as early–mid-Campanian. Plant bearing beds in this area interfinger with marine units allowing precise age determination based on marine moluscs. This flora is typical for the Barykov phase of floral development in the Northern Pacific Region and provides data crucial for better understanding angiosperm evolution in northern high latitudes during one of the warmest time intervals in Earth history. The Barykov
Flora is very diverse (>70 taxa) and is characterised by a predominance of angiosperms (>50%), followed by conifers (17%) and ferns (12%); horsetails, ginkgoes and cycadophytes are less numerous. Angiosperms are the most numerous in all the Barykov plant localities and consist of 38 leaf morphotypes. Among them platanoids
( Arthollia , Paraprotophyllum ), Barykovia tchucotica and several species of Macclintockia are abundant.
Lobed leaves of Cissites beljaevii , C. pekulneensis,
Araliaephyllum , as well as entire margined
Magnoliaephyllum and Juglandiphyllites are characteristic plants of this flora. Two species of
Trochodendroides, Viburniphyllum, Menispermites,
Celastrinites, 'Vitis' penzhinica and Quereuxia angulata are less numerous. The diversity of woody dicot leaves allows us to reconstruct the climate experienced by this flora. To do this we have used the Climate Leaf Analysis
Multivariate Program (CLAMP) method of calculating quantitative palaeoclimate variables. Our estimates using
CLAMP with the PHYSG3BR physiognomic training set
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and a new global gridded climate calibration suggested the mean annual temperature is estimated to have been
+11.2 ± 2.2°C, the warm month mean temperature +20.3
± 2.8°C, the cold month mean temperature +2.5 ± 3.8°C, the mean growing season precipitation 766 ± 392 mm, the mean monthly growing season precipitation 108 ± 52 mm, the precipitation during three consecutive wettest months 558 ± 276 mm, the precipitation during the three consecutive driest months 167 ± 64 mm and the growing season length 6.7 ± 1.4 months. Therefore, the Barykov
Flora existed in a humid temperate climate with warm summers, mild frost-free winters and seasonal variation in precipitation. This flora reflects deciduous predominantly broad-leaved – coniferous vegetation of coastal plains existed at palaeolatitude of approximately
72°N. The deciduousness of the flora was most probably due to not only temperature, but also sunlight seasonality typical for high latitudes.
Tracking validated angiosperm fossil biomolecules
1 through the Mesozoic
Taylor, D 1 , Zinniker, D
Barbanti, SM 4
2 , McCorkle, E
, Moldowan, JM 2
1 , Hu, S 3 ,
2
Dept of Biology, Indiana University Southeast, USA;
Stanford University, USA; 3 Yale University, USA; 4 IPEX
Co
A growing number of molecular fossils are common and abundant in many Cretaceous and Tertiary sediments, but rare in older sediments. The observed spatial and temporal distribution of the biomarker oleanane (and the widespread occurrence of functionalized oleanoids in living monocots and eudicots) has led to oleanane’s use as a qualitative indicator of angiosperm input in sediments. Other potential molecules showing similar patterns include des-A-oleanane (a putative byproduct of oleanane), and bicadinanes (known from dimerization and diagenesis of sesquiterpenoids in resinites from
Dipterocarpaceae). However, interpretations of oleanane occurrence are limited by incomplete taxonomic surveys for natural products in living angiosperms and other seed plants. To examine biomarker distribution at the base of the angiosperm tree, we sampled species from the three most basal orders, Amborellales, Nymphaeales and
Austrobaileyales, as well as Chloranthales, Magnoliales,
Laurales, Piperales, Acorales, Alismatales,
Ceratophyllales and Ranunculales. The samples were subjected to hydrous pyrolysis, extracted, hydrogenated using an ionic reduction procedure, and separated into saturate and aromatic fractions. The presence of appropriate functionalized natural products was tested using GCMS and GC-MRM-MS techniques. The procedure was designed to mimic conditions of diagenesis and burial that transform diverse functionalized natural products into a limited number of identifiable molecular fossils. Oleanoids were found in the most basal orders and a parsimony reconstruction supports the hypothesis that they are ancestral to angiosperms. The two diagenetic transformation products, oleanane and des-A-oleanane, have similar distributions but des-A-oleanane is more easily measured in plant pyrolysates. This distribution is not surprising as oleanane and des-A-oleanane occurrence have been correlated (correlation coefficient 1.0) in a 100+ oilsample set derived from source rocks of a wide variety of
192 geologic ages and depositional environments. Lastly, basal angiosperms do not seem to be a direct source for bicadinanes, supporting previous work showing low correlation to oleanane source rock occurrence, and suggesting they are usually the result of diagenesis and restructuring of existing sesquiterpenoids. Examination of different sediments show the abundance of the molecular fossils appears dependent on the depositional environment. Although this may be related to diagenesis including oxidation, relative comparison based on ratios to hopane (a bacterial biomarker) may show where angiosperms were dominant. Molecular fossil relative abundances are correlated to detailed macrofossil study of the depositional environment at the Aptian Dutch Gap locality in VA, USA that suggest angiosperms were confined to the levee deposits and not a significant contributor to the more diverse flora in the backswamp.
If supported it would suggest angiosperms were initially adapted to unstable sites along stream margins. In addition, the examination of potential angiosperm sister groups such as Bennetittales, Corystospermales,
Glossopterids, Caytoniales and Gigantopteridales for these molecular fossils can indicate which clades may be more likely to be related to angiosperms.
Early angiosperms from the Late Aptian of northern
Gondwana (Crato Formation, Brazil)
Coiffard, C 1
MER 2
, Mohr, BAR 1 , Bernardes-De-Oliviera,
1 Museum of Natural History, Berlin, Germany; 2 Institute of Geosciences, University of São Paulo, Brazil, and
Laboratory of Geosciences, University Guarulhos, São
Paulo, Brazil
Megafossils are the main source for understanding evolutionary patterns in radiation and development of major angiosperm clades because they tell us the story of the tremendous systematic but also morphological and ecological diversification of angiosperms. The Lower
Cretaceous angiosperm megafossil record is scarce, especially in the low latitudes of Northern Gondwana and fertile parts (fruits and flowers) in connection with vegetative parts (leaves, twigs) are exceptional. In this context, the fossil flora of the Crato Formation (Brazil) is a unique snapshot of the Lower Cretaceous
Aptian/Albian vegetation of Northern Gondwana. This flora already shows the four major groups of angiosperms that we know today, i.e. ANITA
( Pluricarpellatia ), Magnoliids ( Araripia , Endressinia ),
Monocots and Eudicots. Several of these taxa exhibit characters that may be interpreted as adaptations to special environments: among the Magnoliales two taxa grew relatively small (semi)coriaceous sheathing leaves.
These characters are today often found in plants that live under (semi)dry and/or seasonally dry conditions.
Furthermore, one monocot taxon seems to have had in its leaves strong sclerenchyma that enabled this plant most likely to prevent a collapse under water deficiency stress.
This feature may point to a life style of some of the early angiosperms as ground cover at partly sunny sites.
However, among Laurales and Eudicots, existed also relatively large lobed (thin) leaves that may have been produced at shaded habitats close to river environments without water stress. Furthermore the Nymphaeales are typical aquatic plants. Overall, the large variety of leaf

size, shape and internal structures of these early angiosperm leaves support the assumption that by the late Aptian flowering plants already grew at different habitats and had expanded in various ecological niches.
Introduction and overview of conifer phylogeny
Rothwell, G 1 , Campbell, C 2 , Escapa, I
, Little, D 7
3 , Gernandt, D 4
, Maps, G 1
,
, Mathews, Holman, G 2
S 8
, Kelch, D
, Stockey, R 5
6
1
3
Ohio University, USA; 2 University of Maine, USA;
CONICET, Museo Paleontológico Egidio Feruglio,
Argentina; 4 5
Canada; 6
UNAM, Mexico;
University of California, Berkeley, USA;
York Botanical Garden, USA; 8
University of Alberta,
7 New
Harvard University, USA
Phylogenetic studies of conifers have achieved progressively finer resolution of relationships through multiple approaches that utilize a combination of traditional and highly sophisticated modern techniques.
An ever increasing body of nucleotide sequence data has provided the basis for resolving relationships among living plants to the species level. The coding of nearly
400 morphological characters for exemplar species of living and extinct species provides an additional new tool for the study of relationships. When a combination of living and extinct conifers is studied, analyses of morphological characters yield resolution of relationships among stem and crown group conifer families, provide calibrations for inferring the ages of living conifer families and genera, and serve as tests for hypotheses of relationships derived from the analysis of nucleotide sequences and rare genetic markers. Although some subsets of the data suggest that different groups of conifers may have undergone parallel evolution from a
Paleozoic grade of coniferophytes, or that gnetophytes may be variously nested among the conifers, a preponderance of evidence from nucleotide sequences, morphological characters, and rare genetic markers supports the hypothesis that conifers form a monophyletic group. Phylogenetic analyses of both nucleotide sequences and morphological characters resolve Pinaceae as the sister to all other families of living conifers. Those analyses also resolve
Podocarpaceae + Araucariaceae as the sister group to
Sciadopitydaceae + (Cupressaceae + the taxads). Both phylogenetic analyses and transformational series of seed cone morphologies reveal changes leading from
Pennsylvanian age walchian Voltziales, through Permian age voltziacean Voltzales, to either Pinaceae, or to
Sciadopitydaceae and Cupressaceae, and resolve tree topologies that generally agree with those from the analysis of living species. These combined approaches strongly suggest that all families of modern conifers have seed cones that are derived from a compound shoot system, and reveal that the extinct Cheirolepidiaceae subtends crown group Pinaceae on the tree. Together these combined approaches yield a much more highly resolved phylogeny for conifers than previously has been possible, and new analytical methodologies both increase our understanding of homoplasy within the data set and help strengthen our confidence in the accuracy tree topology.
Conifer phylogeny based on molecular evidence: taxon sampling, gene sampling and rooting problems
Escapa, I 1 , Catalano, S 2
1 CONICET, Museo Paleontologico Egidio Feruglio,
Argentina; 2 CONICET, Fundacion Miguel Lillo,
Argentina
Phylogenetic analyses of gymosperms in general, and conifers in particular, have been primarily based on DNA sequences. In comparison, studies including morphological information and fossil species as terminals are extremely rare. Molecular phylogenetic analyses have resulted in notorious disagreements about phylogenetic relationships at the family and generic levels, as well as in the rooting of these groups. Most previous analyses are based on limited gene sampling and taxon sampling, which may explain some of the differences in results. Empirical and theoretical studies have repeatedly demonstrated the advantages of extensive taxon and gene sampling in phylogenetic studies, although the complexity of the analysis increases linearly with the addition of genes and exponentially with the addition of taxa. Until recently, computational limits and data availability have limited the number of phylogenetic studies including multiple genes and hundreds of species. However, development and implementation of new algorithms now allow for the analysis of large data sets using parsimony with a reasonable low computational cost. In the present study we gathered a dataset including more than 350 conifer species as ingroup taxa. Outgroup sampling includes species from other groups of extant seed plants Gene sampling includes more than 20 loci. As with many other applications of a supermatrix, ours has a high percentage of missing entries. The results show, in general terms, the monophyly of modern conifer families and other gymnosperm groups, with moderate to high support for numerous relevant clades. In order to test the effect of limited taxon and gene sampling on the results of previous phylogenetic studies we conducted several different exploratory analyses. First, in order to determine the effect of gene sampling on the monyphyly and rooting of various clades a variable number of genes was randomly deleted from the complete matrix. Similar analyses were carried out to test the effect of ingroup and outgroup sampling on the resulting phylogenetic hypotheses. Our results indicate that rooting and topology of ingroup taxa converge to the same result with increasing numbers of taxa (ingroup and outgroup) and genes. This suggests that differences among the results of previous molecular studies of conifers are most likely caused by limited taxon and gene sampling. Even when general topology among extant species can be stabilized using extensive gene and taxa sampling, there are still particular clades that show either rooting or topological problems. This is not surprising given that living species represent just a small fraction of conifer diversity known to have existed since the origins of the group (approximately 320 mya), and therefore a inclusive molecular analysis is just a first step for understanding the evolution of the group. Future studies are likely to resolve these issues by the use of a total evidence
193

approach, including different sources of data (e.g., morphological characters) and fossil species.
Simultaneous analysis of molecules and morphology and the relationship of fossil ovulate cones to living genera of Pinaceae
Gernandt, D
Cronn, R 4
1 , Holman, G
, Liston, A 3
2 , Parks, M
, Rothwell, G
3 , Campbell, C
5 , Stockey, R 6
2 ,
1 Instituto de Biologia, Universidad Nacional Autonoma de Mexico, Mexico; 2 School of Biology & Ecology,
University of Maine, USA; 3 Dept of Botany and Plant
Pathology, Oregon State University, USA; 4 Pacific
5
Northwest Research Station, USDA Forest Service, USA;
Environmental and Plant Biology, Ohio University,
USA; 6 Dept of Biological Sciences, University of Alberta,
Canada
Although Pinaceae has more extant species than any other gymnosperm family and is of great ecological and economic importance, its early evolutionary history is poorly understood. Anatomically preserved pinaceous ovulate cones establish that the family was phylogenetically diverse during the Early Cretaceous.
However, the only extant genus represented by ovulate cones from the Cretaceous is Pinus, the remaining fossil cones are assigned to one of three organ genera. Thirtyfive species have been described for these organ genera from fossils ranging from the Paleogene to the
Cretaceous. PseudoAraucaria (six species) and
Obirastrobus (two species) are hypothesized as monophyletic while the third, Pityostrobus (27 species), is polyphyletic. To investigate the relationship among organ genera and extant species we performed a simultaneous analysis of 40 fossil cones, 20 extant species representing the phylogenetic diversity of
Pinaceae, and three outgroups. Fifty cone characters were included for all fossil and extant taxa while 60 vegetative, pollen, and embryological characters, and 18 kb of DNA sequence from plastid exons were included for extant taxa only. The resulting tree confirms the placement of PseudoAraucaria as closely related to
Abies or Cedrus , is equivocal regarding the position of
Obirastrobus , and confirms that Pityostrobus is polyphyletic but includes many close relatives of Pinus .
Mapping morphological characters over the combined tree permits us to identify those that are most homoplasious, and through a process of reciprocal illumination, to frame and test whether different coding schemes increase congruence.
Molecular phylogenetics of Podocarpaceae: leaf evolution, biogeography and the angiosperm radiation.
Biffin, E 1 , Lowe, A 1
1 The University of Adelaide, Australia
The ecological radiation of the angiosperms has been linked with sharp declines in diversity of non-flowering seed plants, and the virtual elimination of conifers from productive tropical environments. An explanatory hypothesis, which has been widely accepted, invokes vegetative growth rate differences and broadly, predicts that the relatively inefficient stem and leaf hydraulics of
194 conifers leads to lower maximum growth rates, placing them at a competitive disadvantage in the ‘regeneration niche’ relative to angiosperms on productive sites. While this hypothesis seems broadly applicable to Northern
Hemisphere conifers (e.g. Pinaceae), the Podocarpaceae, a predominantly Southern Hemisphere family, is enigmatic, attaining its highest species richness within angiosperm-dominated tropical forests and is arguably the most successful conifer family within these settings.
Understanding the origins of this pattern could shed light, more generally, on the nature and evolutionary dynamics of conifer-angiosperm competitive interactions. A remarkable aspect of the Podocarpaceae is the broad array of leaf morphologies ranging from scale-like and needle-like to functionally broad leaves. Several distinct lineages of Podocarpaceae develop bilaterally or bifacially flattened leaves, cladodes, and short shoots comprising distichously arranged needles or broad leaves analogous to the compound leaves of angiosperms. Leaf flattening in conifers is associated with high photosynthetic efficiency under low light conditions and a plausible hypothesis links the relative success of
Podocarpaceae in productive, angiosperm dominated environments with a high degree of shade tolerance as a consequence of leaf and shoot flattening. As angiosperms increased in ecological importance from the Late
Cretaceous, changing light environments in the
‘regeneration niche’ may have favoured shade tolerant leaf and shoot architecture amongst conifers or alternatively, forced them into marginal habitats where angiosperm competition is reduced. To test these ideas, we develop a family-wide molecular phylogeny for
Podocarpaceae using a Bayesian relaxed clock approach incorporating fossil time constraints. We find the repeated origin of flattened foliage types in
Podocarpaceae, and that these lineages have diversified predominantly through the Cainozoic, and therefore, in the company of canopy forming angiosperms. We consider the tempo of lineage accumulation, and find that the onset of a sustained broad leaved podocarp diversification is coincident with both a decline in the rate of diversification of imbricate leaved lineages and with the onset of the angiosperm radiation. Furthermore, there is a strong evolutionary correlation amongst leaf morphologies and environmental productivity, with broad leaved lineages preferentially evolving tropical distributions. Taken together, our findings support the view that angiosperm-conifer competitive interactions have been mediated at the foliar level, and these have defined broad-scale contemporary phytogeographic patterns.
Cupressaceae: morphology, molecules, and phylogeny
Little, D 1
1 The New York Botanical Garden, USA
I aim to summarize our current knowledge of
Cupressaceae s.l. (including Taxodiaceae and
Sciadopitys) evolution and the expression of that knowledge through botanical nomenclature. A number of organismal (anatomy, biochemistry, micromorphology, reproductive development, reproductive morphology, and vegetative morphology) and DNA sequence characters can be used to reconstruct the evolutionary history of this clade. Of particular interest are those characteristics that

are generally well preserved in fossils and/or those that may be of adaptive significance. For example, leaf architecture (the evolution of light tolerance) and leaf the structure of transfusion tissue (the evolution of drought tolerance). The degree of elaboration around leaf transfusion tracheid pits has been used as a taxonomic character within Cupressaceae. A survey of pit structure resulted in several possible cladistic codings. A series of phylogenetic analyses of morphological, anatomical, and
DNA sequence data along with different codings of transfusion pit structure indicate that pitting characteristics are consistent with the inferred phylogeny to some degree, but these characteristics vary such that they cannot be used exclusively to delimit genera.
Correlations among the size of vermiform thickenings, the occurrence of anastomosis between intracellular pits, and environmental factors (i.e., the amount of precipitation, seasonality of rainfall, and edaphic factors) will be discussed.
1
The Cretaceous Period – the time interval of radiation of modern conifer genera
Kunzmann, L 1 , Mai, DH 2
Senckenberg Natural History Collections Dresden,
Germany; 2 Natural History Museum Berlin, Germany
The Cretaceous period was fundamental for the evolution and diversification of modern conifers. Conifers were major constituents of most vegetation types during the
Mesozoic. Even when angiosperms played a key role in
Earth’s vegetation from the late Early Cretaceous onwards conifers have partly kept their evolutionary potential. Knowledge of Cretaceous conifers, their taxonomy and systematics, their palaeobiogeographic pattern and palaeoecological positions seem to be crucial for understanding and interpretation the extant diversity and phylogenetic trees based on molecular data. Many well-preserved conifer fossils studied could be excellently used as proxy data for molecular clocks.
After a certain decrease and/or disappearance of typical
Mesozoic groups (i.e. Araucariaceae, extinct
Cheirolepidiaceae, extinct Miroviaceae, extinct
Podozamitaceae and others) a mid-Cretaceous radiation has led to a renewal among this clade of seed plants.
Even new families such as Geinitziaceae and
Doliostrobaceae have been evolved. A remarkable diversification in Pinaceae and in Cupressaceae is recognized in the mid to late Cretaceous and much of the modern genera have their oldest fossil record (first appearance) during this time interval. In some groups the occurrence of extinct 'intermediate' taxa (e. g. extinct genera Austrosequoia, Quasisequoia, Habarosequoia and others of the Sequoioideae) show that several conifer clades were probably much more diversified than during the Cenozoic. Cretaceous conifers have been recognized from nearly all palaeo-latitudes, in all palaeoclimatic belts and in all palaeoecological positions. In particular, deciduous taxa such as Metasequoia, Taxodium and
Glyptostrobus (all Cupressaceae) were common elements in the forests of the high Arctic during the late
Cretaceous. They have been coexisted with evergreen
Cupressaceae, Pinaceae and Taxaceae. In the mid latitudes of both hemispheres and in the palaeoequatorial regions members of Cheirolepidiaceae were very common. Although they have often been considered as typical constituents of brackish environments of coastal plains under semiarid to arid climates they also occupied mesic areas under humid conditions. Most remarkable among this family are the frenelopsids with their xeromorphic adaptations and enigmatic forms such as
Glenrosa and Sedites that have leaves with communal stomatal chambers completely unknown from other gymnosperms. The palaeogeographic situation in the late
Jurassic and early Cretaceous is responsible for the occurrence of several families and genera in both hemispheres, e. g. Lindleycladus (Podozamitaceae),
Frenelopsis (Cheirolepidiaceae), and Araucaria
(Araucariaceae). Subsequently the break-up of the
Pangaea, in particular of the southern supercontinent
Gondwana, has nearly prevented a cosmopolitan occurrence of genera that have been evolved during the
Cretaceous (e.g. Agathis (Araucariaceae), Geinitzia
(Geinitziaceae), Quasisequoia (Cupressaceae).
Hard or soft polytomy: phylogeny and evolution of the fern genus
Polystichum
(Dryopteridaceae) in karst
1 caves
Zhang, L 1
Missouri Botanical Garden, USA
One of the most exciting examples of apparently rapid speciation in ferns is the genus Polystichum
(Dryopteridaceae). As one of the largest genera of ferns,
Polystichum contains about 400 species. This genus is nearly cosmopolitan, but the species are most numerous in subtropical regions, with the highest diversity clearly in Asia, especially in southwestern China and adjacent regions. While more than half of the ca. 400 species evolved recently as result of fast radiation, little is known about the species of Polystichum in limestone areas, especially in karst caves, in southern China. There are two competing hypotheses about the ages of cave species: old relics among different caves vs. young radiation. Molecular data placed cave species in younger lineages suggesting that they are new endemics. Most notably, several morphologically and palynologically different cave species formed well-supported monophyletic polytomies. These species occur geographically in adjacent areas. Are these hard or soft polytomies? More molecular data will be added to answer this question. Molecular data also help identify some cryptic species unrecognizable using morphological data only.
Evolution of Sino–Vietnamese limestone cave
Begonia
1
Chung, K-F 1 , Lin, C-Y
Y 3 , Peng, C-I 2
1 , Leong, W-C 1 , Ku, S-M 2 , Liu,
School of Forestry and Resource Conservation,
National Taiwan University, Taipei; 2 Herbarium
(HAST), Biodiversity Research Center, Academia Sinica,
Taipei; 3 Guangxi Institute of Botany, Guangxi Zhuangzu
Autonomous Region and the Chinese Academy of
Sciences, China
195

The regions bordering northern Vietnam and southern
China are dominated by vast limestone terrains. Each year millions of tourists are attracted to this area to enjoy the Chinese watercolor-like scenery of karst topography.
Hidden behind these picturesque landscapes, however, also abounds in rich and spectacular flora comparable to species radiations better known and characterized in remote oceanic islands and tropical high mountains.
Begonia , the sixth most species-rich (ca. 1,600 species worldwide) flowering plant genus, is one of the most remarkable and conspicuous examples of the Sino–
Vietnamese limestone flora. The Sino–Vietnamese limestone Begonia group includes ca. 70 species of sect.
Coelocentrum , sect. Leprosae , and some species traditionally assigned to sect. Diploclinium and sect.
Petermannia . Based on molecular phylogenetic analyses of Asian Begonia , we demonstrated that these limestone
Begonia formed a strongly supported clade regardless of their traditionally infrageneric classification, suggesting a single invading event into the limestone habitats.
Ecologically these Begonia species are confined to various microhabitats of limestone caves or crevices, suggesting a strong niche conservatism to these specialized habitats. With very few exceptions, most caves are inhabited by a single Begonia species, resulting in a pattern of very low alpha (local) but yet very high beta (among different limestone caves) Begonia diversity across the limestone landscape. Morphologically these species are differentiated mainly in vegetative traits (i.e., habit, leaves, rhizomes) of no obvious adaptive value, suggesting that radiation of limestone Begonia is likely non-adaptive. Because begonias are known as poor dispersers and mainly evolved via geographical isolation, we hypothesized that the continuous weathering of limestone landscape and diminishing of cave habitats had accelerated isolation of Begonia populations, facilitating their population differentiation and eventual speciation.
To test this proposition, we investigated correlation between geographic and genetic distances of sampled populations and species to test the pattern of isolationby-distance (IBD) as predicted by the geographic mode of species. Our data identified several instances of strong correlation between the two distances, partially supported our hypothesis.
Botanic diversity in limestone caves of Yunnan–
Guizhou–Guangxi (especially Guangxi), China
Yigang, W 1 , Fang, Wen 1 , Alex, M 2
1 Herbarium, Guangxi Institute of Botany, China; of Botany, Nature History Museum
2 Dept
The vast karst topography in China is located in
Guangxi, Yunnan and Guizhou. It forms a part of the
Dian–Qian–Gui (Yunnan–Guizhou–Guangxi) flora in the flora of China. The Dian–Qian–Gui flora includes 6276 seed-bearing plant species in 248 families and 1454 genera, and can be considered a biodiversity hotspot.
There are a large number of karst caves with large mouth, great depth, and constant humidity in this area.
Many of the species found in caves are endemic species that can be found only in karst caves, and a significant proportion of these are single-point endemics. Take the case of Elatostema (Urticaceae), we had been studying plant diversity in karst caves in the Yunnan-Guizhou-
Guangxi region, have accumulated a large amount of
196 data and materials. In 2010 we received a grant from the
UK's Royal Society to help study the Systematics and biogeography of cave-dwelling Elatostema (Urticaceae).
10 undescribed new species of Elatostema were discovered from the exploration of 25 caves. DNA barcoding has become one of hotspots of biodiversity research in the last five years. In plants, the proposed chloroplast barcoding regions mainly include five coding
( rpo B, rpo Cl, mat K, rbc L, UPA) and three non-coding
(trn Hpsb A, atp Fatp H, psb Kpsb 1) regions. In addition, nrITS is also suggested as a potential plant barcode.
Limited by the universality and resolvability of single barcoding region, five combinations of these regions are proposed at present. We will filtrate and evaluate the most suitable barcoding regions or their combinations for some key, variable and complicated families or genera in limestone caves from Yunnan–Guizhou–Guangxi and
Indo-China Peninsula, in the near future, for instance
Urticaceae, Gesneriaceae, Begoniaceae, Elatostema ,
Pilea , Chirita , Begonia and so on. The aims of our research are to develop and test hypotheses about the origin, evolution and conservation of this unusual and poorly studied flora. We present a summary of our knowledge on the Urticaceae, Gesneriaceae and
Begoniaceae from karst caves. Urticaceae: 108
Elatostema are recorded from Guangxi. Thirty-seven species, ca. 1/3 of that diversity are found in caves. Some are endemic to caves, such as Elatostema astrostratum,
E. bamaense, E. jingxiense, E. gyrocephalum, E. microcarpum, E. recurianum, E. hezhouense, E. tenuinerve, etc. Further investigation into the Elatostema of karst caves and research into the origin and biogeography of this genus will continue to be published.
Gesneriaceae: 41 genera and 202 species are recorded from Guangxi. According to the on-site investigation of ours, 31 species are cave plants, ca. 15.35% in this family in Guangxi. Eight species only can live in karst caves, such as Wentsaiboea renifolia (only species in
Wentsaiboea ). It is an endemic genus of Guangxi, only living in two karst caves in Du'an county. Paraboea velutina (an extremely special species in morphology so that it firstly was described as a new genus). It only can be found in two limestone caves in Fengshan county).
Chiritopsis repanda var. guilinensis ( rare one with strong fragrance in Gesneriaceae), et al. Begoniaceae: 82 known species in Begoniaceae had been confirmed in
Guangxi; twelve species live in karst caves, ca. 14.63% in this family in Guangxi. Eight are endemic species only found in caves, namely Begonia aurantiflora (t is a new species, which was formally released in 2008; its orange flowers are wonderful), Begonia retinervia (its leaf blades usually have beautiful white veins, so that it can be developed as an excellent gardening material),
Begonia picturata ( it is a traditional herbal medicine that works wonders in Jingxi county), et al.
The problem of large genera related to structure of tribes in Apocynaceae
1
Liede-Schumann, S 1 , Endress, M
University of Bayreuth, Germany
1

Starting with the Apocynaceae meeting during the XVI
IBC in St Louis, clade structure of the family has been increasingly resolved, which has brought with it the need to recircumscribe generic limits of a number of taxa. Of the 373 genera currently recognized, a considerable imbalance of species attribution was found: a large number (232 genera; 61.9%) are very small (1–5 spp.).
While a fair number of the small genera are likely artificial, having been recognized as distinct due to some aberrant morphological features and await merging with one of the large assemblages, the number of actual small genera is still disproportionately high. Only 119 genera
(31.9%) are medium-sized (6–50 spp.), and 23 genera
(6.2 %) are large (>50 spp.) to very large (>100 spp.), with Matelea , Ceropegia and Hoya (all ca. 300 spp.) constituting the largest alliances. These 23 large to very large genera comprise some 2812 spp., 75% of which are contained within 15 genera in the crown clade
(Secamonoideae + Asclepiadoideae), which suggests rampant recent speciation and that this was triggered by the advent of pollinia, a key evolutionary event in the family. Other driving forces of speciation appear to be linked with dispersal (e.g. the evolution of arils in
Tabernaemontana ) or pollinator specialization (e.g. the broad range of flower types in Mandevilla ). Large assemblages are found on all continents, and occur in all subfamilies except Periplocoideae. The latter subfamily is restricted to the Old World and possesses an extremely high proportion of very small genera (76%) and only two genera ( Raphionacme and Pentopetia ) exceeding 20 spp.
Only four genera are pantropically distributed, interestingly, two of them among the earliest branching clades of the family. Old World – New World disjunctions are likewise scarce, only occuring in 3 genera of early branching clades. The typical problems of large genera – insufficient taxonomic knowledge, unclear species limits, insufficient sampling of the rarer species, low interspecific resolution for most of the common molecular markers, morphological parallelisms – will be illustrated using the example of Tylophora (Asclepiadeae
– Tylophorinae). Although several lineages within
Tylophora are well supported by molecular data, these neither correspond to previous generic and subgeneric limits, nor do they exhibit a clear morphological or distributional pattern. The possibilities of translating such a phylogeny into a workable classification will be discussed. Despite the serious lack of basic data, several of the large to very large genera are presently under study or have recently been studied with either morphological or molecular methods, or both. The present symposium asks to what extent the radiations in different subfamilies and on different continents do show common patterns concerning timescale, species distribution, morphological and anatomical adaptations, and coevolution with pollinators and herbivores.
New insights on the biogeography and character evolution of the Vinceae – Willughbeieae –
Tabernaemontaneae clade (Apocynaceae,
Rauvolfioideae)
Simões, A 1
Kinoshita, L 2
, Costa, D 1
, Endress, M 3
, Ferraz, C 1
, Conti, E 3
, Morokawa, R 2 ,
1 University of São Paulo, São Paulo, Brazil; of Campinas, Campinas, Brazil; 3
2 University
Institute of Systematic
Botany, Zurich, Switzerland
Tabernaemontaneae, Vinceae and Willughbeieae, as currently circumscribed, comprise 42 genera and about
300 species that form a strongly supported clade
(ViWiTa clade) within Apocynaceae. These tribes are of particular interest for evolutionary and biogeographic studies in the family due to their morphological variation in reproductive traits, and broad geographic range in the tropics and subtropics. More than 650 sequences from about 200 species from the three tribes were integrated in a series of phylogenetic analyses to evaluate relationships at tribal and generic level, and to test both evolutionary and biogeographic hypotheses. The monophyly of the
ViWiTa clade and of each individual tribe is strongly supported, even though relationships among them remain uncertain. An overall correspondence between clade composition and geographic areas suggests simultaneous events of basal splits into paleotropical and neotropical lineages in all three tribes and subsequent splits within these main areas. Results from ancestral area reconstruction were inconclusive for the ancestors of
Tabernaemontaneae and Vinceae clades. A neotropical or paleotropical origin are equally likely hypotheses in both cases. On the other hand, a neotropical origin of
Willughbeieae was reconstructed with a high support due to the basal position of the Hancornia – Parahancornia clade and the sister relationship of the neotropical genus
Pacouria to all sampled paleotropical genera of
Willughbeieae. The evolution of dehiscent follicles with arillate seeds was a major achievement in
Tabernaemontaneae, and is possibly one of the factors that promoted the diversification of the tribe in the tropics. The evolution of drupaceous fruits, a morphologically specialized style-head with a heterogeneous coverage of secretory trichomes and a basal membranaceous ring are putative key innovations in Vinceae. A putative shift to a lianescent habit from tree-like and shrubby ancestors may have favored the diversification of Willughbeieae in the paleotropics, particularly in Africa. Other traits with potential evolutionary value in the ViWiTa clade are the presence and organization of secretory cells at the calyx lobes, presence and morphology of floral nectaries and the ontogeny and composition of fleshy tissues in fruits.
Putting milkweeds (Apocynaceae subfamilies
Asclepiadoideae and Secamonoideae) in context: understanding the origins of extreme pollen transfer
1 efficiency
Livshultz, T 1
The Academy of Natural Sciences of Philadelphia, USA
The milkweeds (Apocynaceae subfamilies
Asclepiadoideae and Secamonoideae) are a species-rich lineage of 3000 species with among the most complex flowers and efficient pollen transfer mechanisms among all flowering plants. Median pollen transfer efficiency
(the percentage of pollen removed from anthers that is deposited on con-specific stigmas) of milkweeds is 20%
(IQR=21.6%, N=37 species) more than an order higher than a 'typical' animal-pollinated angiosperm species
(median=0.64%, IQR=1.61%, N=10 species). I present an integrative hypothesis that proposes an ecological context and an adaptive explanation for the evolution of such extreme pollen transfer efficiency. It is proposed that milkweeds are descended from large woody
197

rainforest vines in Africa that adapted to drier habitats during aridification at the Oligocene/Miocene boundary.
Their extremely efficient pollen transfer mechanism is an adaptation that allowed their ancestors to reproduce under conditions of much reduced population sizes and depauperate pollinator communities experienced during the droughts that accompanied conversion of rainforest to dry forest. This hypothesis is motivated by the contrast between milkweeds (worldwide distribution, abundant in dry woodlands, grasslands, and deserts, smallish vines, erect herbs, geophytes, and succulents with pollen aggregated into pollinia) and their sister group,
Apocynaceae tribe Baisseeae (29 species, predominantly in the Guineo–Congolian rainforest of Central and
Western Africa, large woody lianas with pollen dispersed as monads). I test this hypothesis using biogeographic reconstruction, fossil-calibrated relaxed molecular clock dating, and comparative climate niche analysis in the context of a phylogeny of milkweeds and their closest relatives in the subfamilies Apocynoideae and
Periplocoideae reconstructed using chloroplast and nuclear loci.
The genus
Hoya
(Apocynaceae) and its contribution to the understanding of the diversification of the
Indomalesian archipelago
, Grimm, G 4 , Forster, PI 2 , Grudinski, M 3 , Wanntorp, L 1
1
Muellner, AN 3
Swedish Museum of Natural History, Dept of
Phanerogamic Botany, Stockholm, Sweden; 2 Queensland
Herbarium Dept of Environment & Resource
3
Management, Brisbane Botanic Gardens, Australia;
Biodiversity and Climate Research Centre (BiK-F) &
4 Goethe University, Frankfurt, Germany; Dept of
Paleobotany, Swedish Museum of Natural History,
Stockholm, Sweden
The origin of the Indomalesian biotas, the extreme biodiversity of the region, abundant endemism, and biogeographic barriers like Wallace's line) have long intrigued biologists. Despite the importance of these barriers, some groups of organisms are widespread in the area, crossing Wallace's Line and these may serve as important tools for testing hypotheses on the origin and structure of biodiversity in the area. Studies based on robust phylogenetic trees, dated using fossils and combined with knowledge on the geological and climatic history have the power to challenge the explanatory hypotheses of vicariance, migration and long-distance dispersal, and can bring about a necessary new synthesis in island biogeography. The species-rich genus Hoya of the milkweed family Apocynaceae, with a predominantly
Indomalesian distribution covers five of the currently recognised World Biodiversity Hotspots (Sundaland,
Wallacea, the East Melanesian Islands, the Philippines, and the Queensland Wet Tropics), and therefore fulfils the prerequisites for functioning as a model plant group for disentangling angiosperm diversification in the
Indomalesian region. Current available phylogenetic hypotheses on Hoya are here presented and used in combination with geological and paleoclimatic hypotheses in order to examine the biogeographic patterns of the genus. The new biogeographic results will serve as a tool to better understand the diversification of flowering plants in the Indomalesian region.
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Milkweeds (
Asclepias
) as a model system: progress and prospects in phylogeny, systematics, and evolution of the largest genus of Apocynaceae
1
Fishbein, M 1
Oklahoma State University, USA
In the broad sense Asclepias L. contains 375–400 species, the largest in Apocynaceae, with centers of diversity in North America and Africa. Circumscription of the genus has been in flux, with narrow circumscriptions favoring restriction of the genus to 130
American species. However, it has been exceedingly difficult find any morphological distinction between the
American and African species. Although several distinctive groups of African species have been accorded generic recognition, this leaves many African species residing in Asclepias , which has been considered a taxonomic limbo. Asclepias is also the most thoroughly studied genus from a variety of other perspectives, including reproductive biology, herbivory and the evolution of defenses, population genetics, and phytochemistry. The 'monarch–milkweed' interaction has served as a model system for the study of the interaction between specialist herbivores and their host, with a particular focus on ability of monarch larvae to sequester chemical toxins that are employed by the butterfly adults in their own defense against predators. As a result of recent and ongoing molecular phylogenetic efforts, the current understanding of some relationships in the genus are resolved, despite evidence of an extraordinarily rapid initial diversification. Recent progress includes evidence for an initial split between African and American clades of Asclepias , suggesting a single colonization of North
America from Africa followed by remarkable morphological convergence between similar species in
Africa and the Americas, and the derivation of all South
American species through a single colonization from
North America. Complete genomic sequencing of the first member of Apocynaceae, Asclepias syriaca , will be completed soon and promises to provide quantum leaps in the resolution of Asclepias phylogeny, understanding the genetics of adaptation, the study of population genetic processes, and unraveling the biogeographic history of milkweeds, among many others fields of study.
I will summarize recent progress in Asclepias systematics, including phylogenetic study using whole chloroplast genomes and multiple, unlinked nuclear loci.
The chloroplast data provide for the first time complete resolution at the species level. However, nuclear loci show conflicting phylogenetic signal indicating that incomplete lineage sorting and introgressive hybridization have played an important role in Asclepias diversification and must be accounted for in phylogenetic studies. I will also demonstrate how systematic research on Asclepias has contributed to the development and testing of theories in evolutionary biology, particularly respect to the coevolution of milkweeds and their herbivores and the evolution of anti-herbivore defenses.
Key results include evidence for contrasting evolutionary trends in Asclepias between a reduction in the potency of resistance to herbivory and an increasing ability to tolerate herbivory, and an initially rapid diversification of the genus that may have been coupled with rapid diversification of defense strategies.

The origin and early evolution of Metastelmatinae
(Asclepiadoideae, Apocynaceae), with attention to the basal genus
Minaria
Rapini, A
Berg, C 1
1 , Ribeiro, PL 1 , Soares e Silva, UC
, Liede-Schumann, S 2
1 , van den
1
2
Universidade Estadual de Feira de Santana, Brazil;
University of Bayreuth, Germany
Phylogenetic studies based on plastid DNA have greatly elucidated relationships in Asclepiadoideae
(Apocynaceae), especially at the subtribal level.
Currently, we know that the neotropical species are distributed into only four lineages; the oldest and largest of them comprises four subtribes: Metastelmatinae,
Oxypetalinae, Gonolobinae, and Orthosiinae.
Phylogenetic molecular studies in the three former groups have shown unresolved relationships and/or low support for basal nodes. The Metastelmatinae comprise
13 neotropical genera and about 260 species. Besides being unresolved at basal nodes, they also present high levels of incongruence between plastid and nuclear data.
Together, these results suggest that this large neotropical lineage was subject of several periods of radiations. Such pulses of rapid diversifications would have favored lineage sorting and short branches (i.e. with few synapomorphies) in particular areas of the tree, making actual phylogenetic relationships hard to detect. Despite of these difficulties, molecular studies confirmed that the genus Ditassa is polyphyletic and revealed an independent lineage of subshrubs with minute leaves that diverged during the early evolution of Metastelmatinae; this group was recently recognized as the genus Minaria .
Most (ca. 75%) of the 21 species of Minaria are endemic to the Espinhaço Range, in Brazil. They occur predominantly in open fields, principally on quartzite substrate of the ‘campos rupestres’; some species are restricted to the outcrops, whereas others are exclusive of sandy or stony soils. In this study, we investigate the phylogenetics and biogeography of Minaria , as well as the genetic diversity and structure of its species, to understand the high levels of microendemics in the genus. Our phylogenetic results put the position of M. polygaloides in question. The species is sister to the rest of the genus according to the plastid data, but is not closely related to the genus according to the nuclear data.
Apart from this species, Minaria is divided in two clades; one comprises two species endemic to small areas in the
Espinhaço Range of the State of Bahia, whereas the other comprises 17 species, 12 being endemic to the Espinhaço
Range, eight of them restricted to small areas in the State of Minas Gerais. The Minaria clade is marked by a basal grade composed of microendemic species, whereas the broadly distributed species are derived. Most
Asclepiadoideae are characterized by several comose seeds per fruit, but some micorendemic species of
Minaria have only one or few seeds without a coma.
These characteristics probably limit their dispersion, but are derived states in the lineage; therefore, they cannot be the only reason of the restricted distribution of these microendemic species, nor the reason of the high rates of endemics in Minaria . The species of Minaria present moderate genetic variability regardless of their distribution. Genetic structure patterns cannot be directly associated with their range, as evidence of regular migration between populations occurs regardless of distance. Therefore, factors other than dispersion capacity must be called to explain the high rates of endemics in Minaria .
1
The rise of reproductive barriers in orchids
Cozzolino, S 1 , Scopece, G 1
Dept Structural and Functional Biology, University
Federico II of Naples, Italy
New species arise when reproductive barriers evolve between formerly interbreeding individuals. In orchids and other flowering plants, reproductive isolation is achieved through a combination of pre- and post-zygotic mechanisms. The understanding of the relative contribution of these mechanisms to speciation is difficult because it requires the identification of those mechanisms that limit gene flow between divergent populations that still lack complete isolation. However, not all the divergence among populations leads to speciation and diverging species continue to accumulate ecological and morphological differences, potentially obscuring the relative contributions of various isolating mechanisms during incipient species formation. Only the combination of multiple approaches on the study of reproductive barriers evolution can help in shed light on the speciation processes of orchids. In comparative studies, pre-zygotic isolation has been found to evolve erratically and faster than post-zygotic isolation, possibly because it is more frequently controlled by few genes of major effect. In contrast, the correlation of strength of post-zygotic barriers with increasing genetic distance may indicate a clock-like and slow evolution. Despite these differences in evolutionary patterns and genetic background, the few studies that directly focused on the first phase of the speciation process in orchids, i.e. on the evolution of incipient barriers that arise within species and are initially polymorphic within/among populations, suggest that both pre- and post-zygotic barriers can play a primary role. Accordingly, interspecific crossing experiments in two groups of terrestrial orchids with different levels of pollinator specialization indicate that groups with high pollinator specificity rely on pre-mating reproductive barriers, whereas groups with weak pollinator specificity show strong evidence for intrinsic post-mating barriers (both pre-zygotic and post-zygotic barriers), including late post-zygotic mechanisms as chromosomal divergence. Contact zones between related orchid species represent the natural arenas where reproductive isolation is exposed to natural selection and offer the unique opportunity to estimate the strength of reproductive barriers and, consequently, the evolutionary role of hybridization. The investigation of these contact zones has shown different scenarios depending on the type and on the strength of isolating barriers at work between species. In contact zones between highly specialized species, interspecific pollen exchange is extremely rare and hybridization only occurs under sporadic circumstances. However, if species boundaries are broken and if the hybrids are attractive to pollinators, the weakness (or absence) of post-zygotic isolating
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mechanisms allows extensive gene introgression and genetic admixture. Differently, in contact zones between generalist species the continuous sharing of pollinators often leads to hybridization but parental gene pools are sheltered from introgression by strong post-zygotic barriers and hybrid zones are mainly characterized by first generation hybrids. Whilst most so far investigated contact zones support these two opposed scenarios, some ongoing studies are showing alternative patterns with specialized species sharing pollinators and being separated by strong post-zygotic barriers and with generalist species showing extensive introgression even in presence of differences in chromosome numbers. In this latter case, however, ecological selection was found playing an important role in keeping species cohesion.
Floral signals, pollinator behavior, and reproductive isolation in orchids
1
Schiestl, F 1
University of Zürich, Switzerland
Pollinator behavior strongly impacts on the strengths and mode of pollinator mediated reproductive isolation in plants. The diversity of pollination systems in orchids incorporates a variety of different types of behavior by pollinators. An important distinction is whether pollinators’ visits to flowers are motivated by mating or foraging behaviour. Mating behaviour is characterized by innate preferences for mating signals, mediated by neuronally hard wired behavioural responses. Sexually deceptive orchids imitate mating signals, typically volatile chemicals, and thus exploit male insects that pollinate flowers through attempted copulations. These plants are primarily isolated prezygotically through ethological pollinator isolation by producing speciesspecific mating signals. In the Australian genus
Chiloglottis , single volatile compounds and dual blends thereof mediate species-specific pollinator attraction. In the European genus Ophrys , bends of multiple compounds in specific proportions are key for pollinator isolation. In pollination systems with foraging behaviour, learning of floral signals is pronounced, as pollinators establish floral constancy in case of rewarding flowers but quickly avoid non-rewarding deceptive flowers.
Thus, food deceptive flowers are characterized by high pollinator sharing, and reproductive isolation is primarily postzygotic. Rewarding orchids such as the European genus Gymnadenia , however, have strong floral isolation despite being less specialised in pollinator attraction. In general, there is no association between absolute (number of pollinator taxa) and relative (pollinator sharing) specialization in orchids. This argues against the common assumption that generalisation in pollination makes floral isolation unlikely.
Pollination through sexual mimicry in Neotropical
Orchidaceae: do they follow the same patterns of temperate and paleotropical taxa?
Singer, R 1
1 Universidade Federal do Rio Grande do Sul, Brazil
Orchid pollination through sexual mimicry is one of the most spectacular plant–insect interactions recorded so
200 far. In essence, orchid flowers of some genera mimic visual, tactile and – most importantly – chemical features of some virgin insect females. Males of these insects emerge first and attempt copulation with the flowers, promoting their pollination. This pollination strategy was first described for the Old World genus Ophrys and, later, for several Australian terrestrial orchids. In these taxa this pollination strategy follows some clear patterns:
1) The plants are strongly seasonal, with a well-defined flowering period, 2) pollinators are solitary and produce only one or a few generations in a year, 3) males and females are produced in similar numbers, and 4) pollination is low and takes place during the brief period prior to the emergence of females. Chemical features are essential in this pollination strategy. Several studies on
European Ophrys spp. and their pollinators have clearly pointed out that the key elements in these interactions are cuticular hydrocarbons (alkanes and alkenes) present in both, unpollinated flowers and insect virgin females. A novel kind of compounds called chiloglottones in important in Australian Orchidaceae. During the last decade, pollination through sexual mimicry has been reported in a few Neotropical Orchidaceae and the reports significantly differ from these of European and
Australian orchids. Trigonidium obtusum and Mormolyca ringens (Maxillariinae) are pollinated by drones of eusocial Meliponina (stingless bees). Mormolyca ringens is pollinated by males of Nannotrigona testaceicornis and males of Scaptotrigona sp. Overall; the pollination process resembles this of Ophrys spp. In Trigonidium obtusum the flowers are erect, funnel-like and devoid of insect-like parts. The bees attempt copulation with the perianth (attractive parts differ among individuals) and get briefly trapped into the floral cavity. Trying to find a way out, bees dislodge or deposit pollinaria. Ongoing chemical studies support that the attraction mechanism is similar to this of Ophrys spp. Homologue alkanes and alkenes have been found in both, the cuticles of orchid flowers and these of pollinator’s respective females. The pollination in Lepanthes spp. is more puzzling.
Lepanthes (Pleurothallidiinae) is a very diverse genus throughout the tropical Americas. Pollinators are male
Sciarid flies that attempt copulation with the flowers and may even deposit spermatophores on them during the process. Remarkably, the flowers are often brightly colored and bear no overall resemblance with its pollinators. Although reports on Neotropical Orchidaceae are still scarce, they suggest some different patterns: 1) these orchids flower during the whole or most part of the year, 2) pollinators can be solitary or social, but several generations are produced in a year, 3) as for the
Meliponine bees, drones are produced in exceedingly higher numbers than fertile females. Very few males will reach a female and there will be always plenty of males to interact with the flowers, 4) consequently, pollination may take place during the whole year and is not restricted to a very specific period, such as in temperate and paleotropical taxa.
Context and consequences of repeated evolution of nectar in the rewardless orchid genus
Disa
Hobbhahn, N 1,2 , Johnson, SD 2 , Harder, LD 1
1 University of Calgary, Canada; 2 University of KwaZulu-
Natal, Pietermaritzburg, South Africa

Nectar is an important attractant and reward for flower visitors and promotes frequent visitation, flower fidelity, and pollination for the plants that produce it. Despite these obvious advantages, some plant species offer neither nectar nor other floral rewards, but instead rely on other species for pollinator attraction and rewards. The flowers of rewardless species are pollinated by visitors that have instinctual preferences or that have learned to associate floral signals (colour, shape, and scent) with reward presence, and effect pollination before realizing their mistake. Two non-exclusive hypotheses have been proposed to explain the occurrence of rewardlessness.
The cross-promotion hypothesis proposes that rewardlessness is a mechanism to manipulate pollinator behaviour to enhance cross-pollination and reduce selfpollination. The resource-limitation hypothesis proposes that rewardlessness enables allocation of nectar resources to other floral functions that may improve plant mating and/or fecundity. Despite these proposed benefits, rewardlessness bears mating costs that must be overcome. In particular, rewardlessness can result in infrequent visitation if pollinators discriminate against deceitful flowers, thereby reducing pollen receipt and export, and seed production. Rewardlessness is particularly common in the Orchidaceae, with approximately one-third of the 19,500 species offering no rewards for their pollinators. In the large southern
African orchid genus Disa (180 species), nectar production has evolved at least seven times from rewardless ancestors. The existence of a well-resolved phylogeny and good information on the pollination ecology of many Disa species makes this genus an ideal study system to examine predictions of the above hypotheses in a phylogenetic framework. For up to 89 Disa species we surveyed (1) traits involved in pollinator attraction, such as display size, flower number and size, presence or absence of nectar, and mimicry of co-flowering rewarding species; (2) traits that determine the energetic costs of flowering, such as flower number and biomass, and investment in nectar sugar, and (3) the consequences of nectar production for plant reproduction, such as visitation, pollen dispersal and transfer efficiency, the amount of self-pollination, and the severity of inbreeding depression. We found little support for the resource-limitation hypothesis. Nectar production consumes only a small fraction of the energy invested in flower production in Disa , and rewarding and rewardless species allocate equally to traits that improve pollinator attraction and reproductive output. Instead, our results are more consistent with the cross-promotion hypothesis. Compared to rewarding species, rewardless species generally export relatively more pollen to more distant mates and therefore outcross more frequently, but at the cost of significantly reduced fruit production due to pollen limitation. In addition to testing the above hypotheses, our extensive and diverse data set enables us to identify the nature and phylogenetic context of changes in investment in pollinator attraction and mating system that accompany the evolution of nectar, and to draw conclusions about the context in which nectar evolved. Our examination of the consequences of derived reward production contributes to understanding the function of rewardlessness in the ancestral, deceitful species.
Drivers of orchid diversification – pollinators, or not?
Smithson, A 1
1 School of Plant Biology, University of Western Australia
& Kings Park and Botanic Gardens, Australia
The Orchidaceae is characterised by high current species diversity, highly modified and diversified floral morphology, and phylogenetic signals of explosive adaptive radiations. The high frequency of orchid pollination mechanisms involving pollinator deception have intrigued biologists since Darwin, but the elucidation of selection pressures favouring the evolution of deception remain controversial. Moreover, causal links between adaptive radiation in orchids and key adaptations to pollinators are frequently made, but more rarely tested. Meta-analyses of three major groups within the Orchidaceae found no association between pollinator deception or nectar reward production and increased or reduced diversification rates. Across the Orchidaceae, there is evidence that pollinator specialization is more ancestral, with later-evolving clades tending to have more generalized pollination. Numerous instances of high clade-specific diversification within the
Orchidaceae may be associated with key evolutionary innovations for pollination, but might also be driven by other factors. Given the lack of link between pollination strategy and higher diversification rates, why therefore do orchids have such modified and diversified floral morphology? I argue that highly skewed reproductive success, especially via male function, is enforced by pollinia-based pollen dispersal, and will likely result in strong pollinator-induced selection. Equally, explosive diversification within the Orchidaceae is just as likely to be influenced by adaptation to ecological niche: alternative hypotheses to pollinator adaptation should not be overlooked. I discuss strategies by which we can test diversification hypotheses comparatively.
Pollinator-driven speciation: an evaluation of the evidence in Australian sexually deceptive orchids
Peakall, R 1 , Griffiths, K 1 , Poldy, J 2 , Barrow, R 2
1 Research School of Biology, The Australian National
University, Canberra, Australia; 2 Research School of
Chemistry, The Australian National University,
Canberra, Australia
It has long been hypothesised that pollinators have played a key role in plant speciation. Yet, despite some evidence for pollinator-driven speciation, we still have much to learn about this potentially important process of speciation. Australian sexually deceptive orchids may offer an ideal system for testing this hypothesis. These orchids lure their wasp pollinators to the flower by emitting semiochemicals that mimic the specific sex pheromone of the wasp. In this talk I will explore the emerging support for pollinators as key drivers of speciation in this study system. In a multidisciplinary study of sexually deceptive Chiloglottis orchids we have evaluated wasp pollinator specificity in the field; identified the novel compounds involved in pollinator attraction; and mapped our chemical findings onto a phylogeny of the orchids. We have shown that pollinator specificity has a strong chemical basis, and confirmed
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that speciation is always associated with pollinator switching, usually underpinned by chemical change.
Evidence from chemical, ecological and genetic analysis has confirmed that there are multiple cryptic species of orchid suggesting speciation is ongoing. In a parallel study of the orchid pollinators, multiple cryptic species have also been uncovered. This hidden diversity has likely enabled, rather than constrained, pollinator-driven speciation in these orchids. I will conclude the talk by identifying gaps in knowledge and explore the implications of our findings for the plausibility of pollinators as key drivers of speciation more broadly.
Process behind the pattern of incongruence: diagnosing hybridisation versus paralogy versus lineage sorting using 454 sequence data
Pfeil, B 1 , de Sousa, FDPST 1 , Doyle, JJ 2 , Oxelman, B 1
1 University of Gothenburg, Sweden;
Ithaca, USA
2 Cornell University,
Natural processes that generate gene tree conflict
(incongruence) have been recognised for nearly four decades, but remained under-appreciated until the 1990s.
These processes include: lineage sorting – when alleles coalesce more deeply than when the species that possess them diverged; introgressive hybridisation – when different species occasionally exchange genes, resulting in the transfer of alleles from one species to another or in the formation of new species with alleles from both parents; paralogy (mistaken orthology) – when gene duplication within a species persists through speciation events, such that differential loss of copies in descendant species results in gene trees that don’t match the species tree. Gene trees can be correctly inferred but differ nonetheless, because of lineage sorting, hybridisation and paralogy. Although attention has been directed to understanding these processes, methods that account for them when inferring a species tree have not dealt with all three processes simultaneously. The most progress has been made to account for lineage sorting alone, by using predictions from coalescent theory that models allele creation, retention and loss in populations. Some progress has been made to add hybridisation as well.
However, no method exists to account for paralogy as well as the other two processes, and be able to either produce a species tree directly, or to filter gene trees in some way so as to be able to use existing methods to make species trees from gene trees. Here we present a proof of concept using simulated data that shows how using linked and unlinked genes could be the key to accounting for all three causes of incongruence simultaneously. We show how genes affected by paralogy can be detected and removed. Then, using existing but recently developed methods, we can filter those gene trees of individuals affected by hybridisation and proceed to inferring the species tree from the remaining gene trees and individuals. We also show how
454 technology may be an efficient way to generate the required multi gene data for these investigations.
202
Estimating the species tree for Hawaiian
Schiedea
(Caryophyllaceae) from multiple loci in the presence of reticulate evolution
Willyard, A 1 , Wallace, LE 2
Sakai, AK 4 , Nepokroeff, M 5
, Wagner, WL 3 , Weller, SG 4 ,
1 Hendrix College, Conway, AR, USA;
University, MS, USA;
Washington, DC, USA;
CA, USA;
USA
5
4
3
2 Mississippi State
Smithsonian Institution,
University of California, Irvine,
University of South Dakota, Vermillion, SD,
Schiedea (Caryophyllaceae) is a monophyletic genus containing 34 species that arose from a single colonization of the Hawaiian Islands. The genus is one of the best examples of adaptive radiation in the Hawaiian
Islands, with species exhibiting a variety of growth forms and dramatic transitions in breeding systems from hermaphroditism toward dimorphism or autogamy. Our study included 91 accessions and 4 independent genetic partitions: 8 plastid and 3 low-copy nuclear loci (9217 bps). Despite incomplete resolution in the shallow tips of each gene and species tree, our results support tests of species monophyly for named taxonomic species. Gene trees revealed cytonuclear incongruence at a deep node as well as among species. We compared: a 3-locus nuclear species tree using BEST; concatenation; and
'total-evidence' (excluding recognizable plastid introgression and incorporating morphological data for each species). Ages inferred from a BEAST analysis allow an original colonization onto either Nihoa or
Kaua`i. We suggest that several hard polytomies on the species tree are biologically realistic, signifying either nearly simultaneous speciation or historical introgressive hybridization. Based on inferred node ages that exceed expected coalescent times, we propose that undetected nuclear introgression may play a role in the incomplete lineage sorting observed in sections Schiedea and
Mononeura .
Detecting and diagnosing incongruence across the genome with short-read transcriptome sequencing:
1 examples from
Glycine
(Leguminosae)
Doyle, J 1
Cornell University, USA
Short-read transcriptome sequencing provides a relatively affordable means of assaying the entire gene space of the genome. As such it should be useful for detecting incongruence across many nuclear loci.
However, to employ short-read technology in evolutionary studies of non-model plants, numerous technical challenges must be overcome, among them the problem of mapping reads to highly duplicated reference genomes. This is a particular hurdle for taxa that have experienced relatively recent whole genome duplications, such as the legume genus, Glycine , which underwent a polyploidy event within the last 10 million years.
Consequently, the fully sequenced soybean ( G. max ) reference genome consists of numerous duplicated blocks whose homoeologous gene pairs are typically >80% similar, making it difficult to map short reads unambiguously to one homoeologue. Our studies focus on the ca. 26 predominantly Australian perennial species

of subg. Glycine , which diverged from the annual, northern Asian subg. Soja (soybean and its wild progenitor, G. soja ) around 5 million years ago. The perennials subsequently diverged into several 'genome groups' of reproductively compatible species that are reproductively isolated from other genome groups.
Incongruence has been detected at various hierarchical levels in the subgenus using the chloroplast genome and a handful of nuclear genes. For example, chloroplast and nuclear genes reveal very different patterns of relationship among genome groups. Most perennial species are inbreeders, but there is also evidence of introgression within genome groups, and the existence of allopolyploids formed between genome groups also attests to the potential for hybridization. Deep coalescence due to lineage sorting is, of course, another source of incongruence, particularly for closely related species. We have generated Illumina transcriptome data as part of several different projects in subg. Glycine , involving both intra- and interspecific comparisons from species representing several genome groups. Thus we are in a position to assay genome-wide patterns of variation, and to assess the degree of incongruence at various taxonomic levels in the group. We will also be able to compare the short-read data with information from BAC sequencing of up to a dozen unlinked 350 kb genomic regions in soybean and orthologues across the perennial group.
Investigating the role of recombination in incongruence among datasets using partition jackknifing
Egan, AN 1 , Doyle, JJ 2
1 East Carolina University, USA;
USA
2 Cornell University,
Subtribe Glycininae (Leguminosae) shares an ancient polyploidy event with many other legumes. The subtribe also contains Glycine , a genus of more recent polyploid origin that includes soybean, G. max (L.) Merr. Our efforts to understand the evolutionary history of subtribe
Glycininae uncovered much phylogenetic incongruence among our nuclear gene regions. We conducted a thorough investigation into the source(s) of phylogenetic discord by screening for recombination and paralogy.
While paralogy/orthology conflation was not deterministically a source of incongruence among our genes, the effects of mistaken paralogy cannot be completely ruled out due to the possibility of undetectable birth-death events. We detected multiple signatures of recombination in our data that were often associated with intron/exon boundaries. To further explore these signatures and parse the distribution of phylogenetic signal in our data, we conducted thorough partition (gene and intron/exon) jackknife analyses and compared topology and nodal support produced via maximum likelihood and maximum parsimony to those resulting from individual genes and combined data. Our jackknife analyses often supported the presence of recombination. Our comparative phylogenetic analyses revealed that one gene was contributing disproportionately to the signal for relationships surrounding Glycine . We illustrate that the joint efforts of individual and combined analyses along with partition jackknifing can provide valuable insights into the distribution and effects of phylogenetic signal and can go above and beyond mere estimation of phylogenetic relationships through the added value of quantifying and qualifying phylogenetic incongruence.
The last frontier: the marine benthic flora of north-
1 western Australia
Huisman, J 1
School of Biological Sciences and Biotechnology, and
Western Australian Herbarium, Perth, Australia
Australia can lay claim to one of the richest and beststudied macroalgal floras in the world, but our understanding of this flora is geographically biased towards the southern and eastern coasts, the most populous regions that have also historically been home to the majority of active phycologists. In comparison, the north-west coast of Australia has been virtually neglected, a perhaps understandable situation given the region’s remoteness and significantly greater logistical difficulties, in addition to huge tidal flows, turbid water, and the presence of several top-level predators! Over the last 10 or so years the author and associates have been opportunistically collecting and intensively studying this marine flora, which has, in part, become considerably more accessible due to the resources boom presently underway in the region. These studies, the results of which will eventually be published in the ‘Algae of
Australia’ series, have revealed a trove of undescribed taxa, as well as numerous poorly understood species previously known from limited historical collections, plus a large contingent of taxa that are common elements of the tropical Indo-West Pacific flora. Many of the collected specimens have also been incorporated into ancillary DNA-based phylogenetic studies, thereby enabling a more global perspective of the north-western
Australian flora. This seminar will include an overview of the broader project, plus a more detailed examination of some of the more interesting taxa that have emerged from the studies.
Isolation and connectivity in the Southern
Hemisphere: phylogenetics targeting different time scales shows that the south has been an area of origin
1 and an area of homogenization
Zuccarello, G 1
Victoria University of Wellington, New Zealand
The Southern Hemisphere has been proposed as a center of origin for much of the present day marine algal diversity. The position of the large Gondwanan land mass in temperate waters has facilitated this diversity.
The low natural dispersal of algal propagules produces populations that are genetic isolated at relatively short spatial scales, this combined with stable environments has lead to speciation events over long periods and over lengthy coastlines. The flora of the Southern Hemisphere
203

has many examples of groups that are unique to the area, and are sister to groups found in more northerly localities. The diversity of many higher taxa (families, genera), is also higher in the Southern Hemisphere. We used phylogenetic analysis to show the high diversity of members of the Fucales in the Southern Hemisphere. We also show that multiple dispersals, and subsequent speciation, is inferred to have occurred to the Northern
Hemisphere. While on an evolutionary time scale the geology and environment of the Southern Hemisphere has lead to high biodiversity of marine algae, more recent oceanographic changes have lead to connectivity across the Southern Hemisphere. The final opening of the
Southern Ocean has led to the formation of the Antarctic
Circumpolar Current, also called the west wind drift.
This continuous strong current has led to many organisms being transported around the Southern
Hemisphere. This is especially true for algae that float.
Rafting and recolonization of southern shores is seen in several algal groups. Research has shown that this has lead to homogenization of much of the southern flora, with little genetic differentiation in areas directly impacted by this current. The strength of this current in shaping genetic connectivity in the Southern Hemisphere is just being explored, and shows the continued fascination of the southern oceans in the evolution of marine algae.
A Gondwanan origin of the brackish green algal genus
Wittrockiella
(Pithophoraceae, Cladophorales)?
Boedeker, C 1 , Nelson, W 2 , Stech, M 1 , Zuccarello, J 3
1 NCB Naturalis /National Herbarium of The
Netherlands; 2 National Institute for Water &
3
Atmospheric Research, Wellington, New Zealand;
Victoria University of Wellington, New Zealand
The apparent Gondwanan distribution of the brackish green algal genus Wittrockiella (Pithophoraceae,
Cladophorales) in the Southern Hemisphere has (once again) raised the issue of vicariance versus long distance dispersal. To address this question some idea of divergence times between lineages is needed. In fossil poor groups such as most algae this is not easy and calibration points are hard to come by. The
Pithophoraceae (= Aegagropila -lineage) is considered ancient (>200 mya) and is an early diverging member within the mainly marine Cladophorales, and is sister to the highly diverse Cladophora - and Siphonocladus lineages. The Pithophoraceae represents an assemblage of five species-poor genera distributed mainly in freshwater and brackish habitats. The current distributions of several taxa of the Pithophoraceae seem to have retained ancient biogeographical patterns, in particular in the genus Wittrockiella . Most species of the
Pithophoraceae appear to be poor dispersers and have restricted distributions, which allows to use geological calibration points. One crucial location is ancient Lake
Baikal (Russia), estimated to be 20–25 my old, which harbours a species assemblage of endemic cladophoralean algae that are members of the
Pithophoraceae. We use DNA sequences of those organisms and the age of Lake Baikal as a calibration point to date the phylogeny of the Pithophoraceae, which in return enables us to discuss the phylogeographic patterns of the genus Wittrockiella with regards to past
204 geological events, including the break-up of Gondwana.
Recent collecting and sequencing efforts have greatly increased our knowledge of the genus Wittrockiella , and have doubled the number of recognized species from three to six. Preliminary phylogeographical data point to an origin of the genus Wittrockiella in the Southern
Hemisphere/Gondwana. Vicariance versus long-distance dispersal scenarios are also tested at the intraspecific level to explain the disjunct distribution of W. lyallii between southern New Zealand and southern Chile.
Out-of-Australia: historical biogeography, diversification and niche shifts of marine macroalgae
De Clerck, O 1 , Verbruggen, H 1
1
Leliaert, F 1
Ghent University, Belgium
, Tyberghein, L 1 ,
The Australasian marine flora has featured prominently in scenarios aiming to explain the present day distribution and diversity of benthic marine macroalgae.
It is often hypothesized that many of the modern algal families have a Mesozoic origin in cold- and cooltemperate waters of the southern hemisphere rather than tropical regions. Support for a southern origin of many algal groups comes from the recognition of possible ancestral taxa in the cold- and cool-temperate shores of
Australia, Tasmania and New Zealand. Tropical regions on the other hand are thought to be dominated by relatively young lineages, having largely diversified in the Paleoceen and Eoceen possibly in response to increased herbivory. Data availability, e.g. inadequate diversity estimates, geographical sampling bias, uncertainty about phylogenetic relationships and divergence times, as well as methodological issues related to historical biogeography, make that scenarios of diversification have rarely been tested explicitly. By integrating parametric models in historical biogeography we test the hypothesis that: 1) independent marine algal flora’s developed along the western and eastern shores of
Australia while it was still part of Gondwana; 2) the
Australian algal flora acted predominantly as a donor region from which species dispersed in the Cenozoic; 3) the diversification of typical tropical families is a relatively recent phenomenon coinciding with thermal stratification of the oceans and increased herbivory. We make use of global algal phylogenetic datasets (e.g.
Dictyotales, Bryopsidales) developed over the last decade in our research group and integrate these data with species distribution models and divergence time estimates to explore the evolution of ecological niches.
Racemose inflorescences in monocots: diversity, development and evolution
Remizowa, MV 1
Rudall, PJ 2
, Choob, VV 1 , Lock, IE 1 , Sokoloff, DD
1 Moscow State University, Moscow, Russia;
Botanic Gardens, Kew, Richmond, UK
2
1 ,
Royal

Analysis of inflorescence diversity in a molecular phylogenetic context suggests that racemose (partial) inflorescences (racemes, spikes, spadices) represent a plesiomorphic condition in monocots. Among earlydivergent monocot lineages, racemose inflorescences are typical for Acorales, several families of Alismatales,
Petrosaviales, some Liliales and some Dioscoreales.
Racemose partial inflorescences characterize several species-rich derived monocot clades, including orchids, sedges and grasses. Several factors cause problems in identification of the inflorescence type in some monocot lineages, including contrasting interpretations of the flower–inflorescence boundary and distinguishing between specialized monochasia and spikelets. In many cases, monocot reproductive structures previously reported to be pseudanthia can be plausibly reinterpreted as flowers of unusual morphology (e.g., Potamogeton,
Triglochin, Centrolepis ). Conversely, the so-called flowers of Zannichelliaceae and Cymodoceaceae
(Alismatales) could indeed be homologous to entire racemose partial inflorescences. They could result from amalgamation of all individual flowers and loss of their boundaries. Dorsiventral spikelets, which are characteristic of some commelinid monocots
( Tradescantia in Commelinaceae; Centrolepis and
Aphelia in Centrolepidaceae), superficially resemble cincinni, a kind of monochasial cymose inflorescences, though detailed investigations strongly support the racemose interpretation. Racemose inflorescences of early-divergent monocots show considerable diversity in terms of presence or absence of flower-subtending bracts as well as in presence, number and position of additional phyllomes on pedicels. These features are of taxonomic and phylogenetic importance and often characterize major clades. They are also significant for understanding monocot flower development. Racemose inflorescences in which flower-subtending bracts are either absent or inconspicuous are common in the basal monocot lineages
Alismatales and Acorales, representing two different patterns of bract reduction: (1) complete bract suppression (the cryptic bract condition) and (2) formation of a ‘hybrid’ organ by overlap of the developmental programs of the flower-subtending bract and the first abaxial organ formed on the floral pedicel.
In the latter case, a flower-subtending bract is absent as a separate organ, but its features are partially expressed in the ‘hybrid’ organ. Different patterns of bract reduction can occur in different species of the same genus. The formation of a flower and its subtending bract from a common primordium and/or retardation of bract initiation could both be regarded as preconditions for bract reduction in Alismatales. The constant occurrence of a bracteole on a flower pedicel is a common feature in many monocots. Spatial insertion of a bracteole is an essential factor regulating sequence of initiation of floral organs. In turn, patterns of bracteole initiation often correlate with phyllotaxy on the main inflorescence axis.
In monocots, racemose inflorescences with bracteoles are closely related to thyrses because the occurrence of nextorder flower in the bracteole axil is a labile feature. The vascular anatomy of the inflorescence axis and the nature of vascular connections between flower and inflorescence will be discussed in several groups. The vascular system of the inflorescence axis and pedicels follows the typical seed plant eustele (or atactostele), but radial divisions of vascular bundles occur in flowers of many monocots. The evolutionary and developmental implications of this important difference will be discussed.
Structural bases for the development of the terminal flower in inflorescences
Bull-Hereñu, K 1 , Classen-Bockhoff, R 1
1 Institut für Spezielle Botanik, Johannes Gutenberg
Universität, Germany
The presence and absence of the terminal flower (TF) in inflorescence represents a matter of wide interest in botany, from its use as a morphological classificatory attribute to the study of the molecular mechanisms that regulate its production. We test the hypothesis whether there are structural prerequisites in the inflorescence meristem (IM) for the TF to be originated. To evaluate this we performed a comparative developmental study with the aid of the scanning electron microscopy, including 20 species of the Apiaceae, Campanulaceae,
Rosaceae, Berberidaceae and Fumariaceae.
Measurements performed in developing IMs show that
TFs appear on IMs which are 2.75 (se = 0.38) times larger than the youngest lateral reproductive primordium.
The shape of these IMs is characterized by a leaf arc
(phyllotactic attribute) of 91.84° (se = 7.32) and a meristematic elevation of 27.93° (se = 5.42). IMs of open inflorescences show a significant lower relative surface, averaging 1.09 (se=0.26) times the youngest primordium size. The relative lower size of open IMs is either a condition throughout the complete ontogeny (‘open I’) or a result from the drastic reduction of the meristematic surface after flower segregation (‘open II’). We conclude that a suitable bulge configuration of the IM is a prerequisite for TF formation in inflorescences and that there are at least two distinct kinds of ontogenies of open inflorescences, suggesting two different causes for terminal flower loss in the course of evolution.
Towards an ontogenetic understanding of inflorescence diversity
Claßen-Bockhoff, R 1 , Bull-Herenu, K 1
1 Institut für Spezielle Botanik, Johannes Gutenberg-
University Mainz, Germany
Though interest in inflorescences has dramatically increased in the last years a consistent classification of their diverse forms is still lacking. Going back to the meristematic origin of inflorescences, their position within the plant body and basic developmental processes we aim to introduce an ontogeny-based reference framework for inflorescences. Inflorescence meristems
(IM) differ from vegetative ones in having a limited activity and being reproductively stimulated by enlargement, reduction of leaf primordium size and axillary bud promotion. They generate reproductive subunits until the formation of flower meristems (FMs), which on their part, directly produce flower organs.
According to this developmental sequence, flowers and inflorescences only differ in the number of fractionating steps; the boundary among them can get indistinct or even lost. Inflorescences are classified after the presence/absence of a terminal flower and the nature of their lateral subunits.
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• Dependent on the formation of a terminal flower,
‘closed’ and ‘open’ inflorescences appear.
Developmental studies indicate that there are two different kinds of open inflorescences (open I, open II) and that the geometrical configuration of the IM is involved in the formation of terminal units. Variation in these meristematic conditions result in facultative terminal flower production as found in many angiosperm lineages.
• IMs usually produce subunits in an acropetal
(‘racemose’) sequence maintaining their meristematic activity at the tip. If the lateral subunits are flowers a
‘simple raceme’ is formed, if they are cymes a `simple thyrse´ results. Cymes are characterized by producing their terminal flower first and maintaining their meristematic potential in the axils of the prephylls. In compound inflorescences the basic pattern is
(multiplicative) repeated resulting in ‘compound racemes’, ‘panicles’ and ‘compound thyrses’. All inflorescences may or may not produce terminal units.
As to their position in the shoot system, reproductive meristems (flowers/inflorescences) are terminal, terminal and lateral or exclusively lateral.
• In the latter case, the apical meristem of the mother shoot remains vegetative and may even continue to grow.
From the ontogenetic view, such a shoot system (called
‘proliferating inflorescence’ in the past) does not represent a single inflorescence but a cluster of several flowers/inflorescences each originating from independent
FMs/IMs.
• These lateral systems flowering in acropetal order contrast with a second set of lateral inflorescences which flower basipetally and only appear below a terminal IM.
Its reproductive stimulus extends downwards to formerly vegetative buds which gradually start to flower.
Dependent on their age they produce a varying number of vegetative leaves before flowering. Contrary to almost all existing inflorescence concepts, position, flowering sequence and leaf shaping are important characters in the ontogenetic classification. They allow recognizing the lack of homology between apparently similar systems, all termed ‘synflorescence’ in the past. The ontogenetic approach illustrates that i) inflorescences can be defined by their meristematic origin, ii) diversity can be reduced to few basic developmental processes, iii) transitional forms can be easily explained by ontogenetic changes and iv, homology statements can be assessed considering position and geometrical conditions of the IM.
Metameric development, positional segregation, and the evolution of distinct floral types within inflorescences
Diggle, PK 1
1 University of Colorado, USA
Architectural effects are defined as regular, repeatable patterns of intra-inflorescence or intra-individual variation in form and/or reproductive potential that are due to flower position. Multiple studies have demonstrated that architectural effects are common among hermaphroditic flowers within inflorescences. I will review the evidence for architectural effects within monoecious taxa, which bear flowers of contrasting sexual function. Positional variation in flower function may have provided 'raw material' for the evolution of
206 unisexual flowers in some lineages, and the commonly observed positional segregation of flower types within the inflorescences of monoecious taxa may be the result of such precursor patterns.
Synergy and interference in the reproductive function of inflorescences
Ishii, H 1 , Kudo, G 2 , Ida, T 3 , Gallwey, J 3 , Harder, L 3
1 University of Toyama, Japan;
Japan; 3
2 Hokkaido University,
University of Calgary, Canada
Although flowers are the essential organs of angiosperm reproduction, the performance of individual flowers reveals an incomplete picture of a plant’s reproductive success, because of diverse consequences associated with the production and functioning of flowers in inflorescences. This inadequacy arises because a plant’s flowers differ in their reproductive potential and phenology and because flowers act together to attract and manipulate pollinators, influence self- and crosspollination, and convert the plant’s resources into seeds.
Conversely, inflorescences are aggregate, rather than unitary, structures, so many of their functional features emerge from the combined characteristics and dynamics of their constituent flowers. With a series of experimental examples, we illustrate how the interplay between floral and inflorescence characteristics influences angiosperm reproduction. Detailed studies of Delphinium glaucum racemes identify diverse aspects of the inflorescence context of floral function. Within inflorescences, flower size, ovule number, female-phase duration, overall floral longevity and anther-dehiscence rate decline from bottom to top flowers, whereas stamen number increases, indicating parallel trends in reproductive capacity. In addition, both the rate at which flowers open and their longevity decline with the time since an inflorescence began flowering, directly affecting the number of flowers displayed simultaneously (display size). Several consequences emerge from these patterns. First, they alter the attractiveness of inflorescences to pollinators, because attraction increases in a decelerating manner with display size and also varies positively, but independently, with flower size. Second, the dynamics of display size and the durations of the male and female phases of the protandrous flowers influence the opportunity for among-flower self-pollination
(geitonogamy). Third, these dynamics govern counterintuitive changes in the gender of an inflorescence and its overall reproductive potential. Together, these influences illustrate the complex, often conflicting, mating processes that contribute to selection on floral and inflorescence characteristics, even in relatively simple architectures such as racemes. To demonstrate some potential effects of alternate arrangements of flowers, we describe how increasing the number of dimensions in which an inflorescence presents flowers alters bumblebee behavior and its mating consequences. Increased dimensionality of artificial inflorescences amplifies the time that nectar-collecting bees spend searching for unvisited flowers, reducing an inflorescence’s profitability and precipitating departure after fewer flower visits. Consequently, if pollinators are abundant, increased inflorescence dimensionality should limit geitonogamy and promote pollen export. The final example illustrates the inflorescence context of resource

allocation among flowers within racemes of Stenanthium occidentale . Overall, cross-pollinated fruits set more seeds per ovule than self-pollinated fruits; however, this effect was accentuated if other fruits on the same inflorescence were cross-pollinated, rather than selfpollinated. In addition, fruits low on inflorescences, and so closer to resource supplies, set more seeds per ovule than upper fruits only if the competing fruits were crosspollinated. Thus, even when inflorescence effects on pollination are equalized, resource interactions associated with the joint production of flowers within inflorescences shape a plant’s reproductive output. Together, this set of examples underscores the reproductive synergy and interference that accompanies the aggregation of flowers into inflorescences. Because of this interdependence, neither floral nor inflorescence characteristics likely evolve in isolation.
Molecular basis of grass inflorescence evolution
Reinheimer, R 1 , Zhong, J 1 , Kellog, E 1
1 Dept of Biology, University of Missouri Saint Louis,
USA
The grass (Poaceae) family includes about 12000 species and over 700 genera segregated into 13 subfamilies.
Grasses are widespread all over the world and their economic importance is undisputable. Some of the family's most well known members are sugar cane, numerous cereal crops such as maize, rice, oat, wheat, etc., and various forage grasses. Grasses are mainly differentiated from the rest of the Angiosperms by their unique flower and inflorescence morphology. Indeed, grass flowers (or florets) do not have conventional sepals and petals, and instead have two to three fleshy organs named lodicules in place of petals, and two empty bracts, presumably known as lemma and palea. One to forty florets, depending on the species, are grouped in novel structures called spikelets, which in turn are located in diverse inflorescence branch systems. The family exhibits remarkable variation in inflorescence morphology. Inflorescences may be lax to condensed, spiral, bilateral, or unilateral, with or without a terminal spikelet, with one to many branches varying in the degree of ramification, length of branch internodes, and disposition, among other traits. Inflorescence diversity in grasses is thought to be caused by modifications of the gene regulation that controls their development.
Genomes and gene duplications, as well as gene loss, are common events in grasses. Duplicated genes can be maintained in the genome for long periods of time mainly by redundancy (maintenance of the ancestral copy by the copies), subfunctionalization (splitting up of the ancestral role by the copies) or neofunctionalization processes (one of the copies acquires a novel role). Given that, variation in when, and where gene copies are transcribed, and how their protein interacts during organ initiation, and development is thought to be the key factor in generating novel forms in grasses. Here we will report advances in the study of genes that play an important role in determining inflorescence diversity in grasses throughout their molecular evolution and modification of their expression patterns over the course of grass family evolution.
Inflorescence and floral structure of the earlydivergent grass
Anomochloa
Sajo, G 1 , Pabon, N 2 , Jardim, J 3 , Stevenson, DW 2 , Rudall,
1
PJ 4
3
Departamento de Botânica, IBUNESP, Rio Claro,
Brazil; 2 New York Botanical Garden, Bronx, USA;
Depto Botânica, Centro de Biociências, UFRN, Natal,
Brazil; 4 Royal Botanic Gardens, Kew, Richmond, UK
The monotypic grass genus Anomochloa belongs to a small subfamily (Anomochlooideae) that is sister to all other grasses. Its reproductive structures are difficult to homologise with those of a typical grass spikelet, in which the glumes and lemmas represent modified leaf sheaths and the palea is a presumed prophyll subtending each flower. We compare the reproductive morphology and anatomy of Anomochloa with 'true' grasses, other early-divergent grasses such as Pharus and
Streptochaeta , and related genera of Poales, such as
Ecdeiocolea . We interpret the outer bract of the
Anomochloa spikelet as a lemma, despite its unusual orientation, in which case the inner bract is a palea. As in a 'typical' spikelet, the palea subtends the flower that is thus subterminal, , and not terminal, as earlier interpreted. The two bracteoles of each ramification are glumes. The single vascularized carpel of Anomochloa is abaxial, in contrast to the three carpels of the sister genus
Streptochaeta and of the other basal genus Pharus (one abaxial and two adaxial carpels). However, we interprete two ventral vascular bundles, found in Anomochloa flowers, as related to missing carpels indicating that in
Anomochloa the gynoecium is pseudomonocarpelar, as are those of Streptochaeta and Pharus . The fourstaminate condition of Anomochloa is unusual within
Poaceae, where three stamens predominate, though four stamens also occur in Ecdeiocolea and some species of
Pharus . Anomochloa is also unusual in that the embryo is protected by the persistent coriaceous palea, in addition to the pericarp and seed coat, whereas in most other
Poaceae the dispersal unit is an indehiscent fruit
(caryopsis) that bears a single seed fused to the pericarp.
Using inflorescence structure to inform character
1 state description: the importance of context
Kirchoff, B 1
University of North Carolina at Greensboro, USA
Many examples show us that the boundaries between the inflorescence and flower are not precisely defined. The most obvious of these are pseudanthia such as are found in the Euphorbiaceae and Compositae. However, even in plants with a more well-defined separation between the inflorescence and flower, there are developmental and structural interdependencies between these units that should be taken into account in any description of flower structure or development. These interdependencies may have a developmental basis, but are also likely to be shaped by selection pressures. Floral orientation is an important parameter influencing plant pollinator interactions, and is thus likely to be under selective pressure. Within the Zingiberales the mature flowers are always oriented so as to facilitate pollination. To accomplish this 'normal' patterns of floral development
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are sometimes modified to change floral orientation from the time of initiation. In other, rarer cases floral orientation is adjusted late in development through movements of the pedestal (resupination), or portions of the ovary. The way in which mature floral orientation is achieved influences the relationship between the flower and inflorescence primordia. In cases where floral orientation changes late in development, early patterns of floral organ initiation often follow those seen in other monocotyledons. In cases where floral orientation is achieved early in development, developmental patterns that affect floral orientation may differ significantly from those found in other taxa. In both cases, floral orientation relative to the inflorescence axis at early stages of development can be an important factor to consider when establishing homologies among developmental stages.
Recent work on visual character description (description not based on verbal characters) has established methods for extracting character data from developmental sequences. These methods are based upon photographs of homologous developmental stages that are sorted into hierarchical groups to represent the phylogenetic relationships among the taxa, based on a specific organ and developmental stage. Using these types of visual methods to establish characters avoids many of the problems that arise from the application of verbal labels.
However, the use of visual character creation methods can be impeded by a failure to consider the context in which they developmental stage occurs. In the case of flower development, this means considering the placement of the flower within the inflorescence. For instance, it is possible to define a developmental stage at which the third sepal (and first pedal) are being formed in the flowers of the Zingiberales. To compare flowers at this stage it is necessary to understand how the flowers are oriented within the inflorescence. Gross similarity among the floral primordia misrepresents the true homologies of the floral organs, and would lead to mistaken conclusions about the phylogenetic relationships among the taxa. Placing the flowers within the developing inflorescence allows for correct homology assessments, and provides a way to extract novel phylogenetic signal from developmental data.
Inflorescences and flowers structure and development in some Celastrales: their correlations with molecular data
Savinov, I 1
1 Moscow State University of Applied Biotechnology,
Russia
One of the most actual problems is the search for morphological characters that may be useful as markers for taxa of different ranks, and their comparison with new molecular data on nuclear and plastid DNA sequences (together with data on plant geography). The author has conducted a complex comparative morphological study of 60 genera and 170 species of the
Celastrales (according to APG III 2009: almost 60 % of genera and 15 % of species from this taxon of the world flora) with a special focus on inflorescences and flower structure and development. There are two big groups of inflorescences in the Celastraceae s. str. – intercalary
(more often) and terminal. For many genera of
Celastraceae one may observe both types of
208 inflorescence, although the frequency of their meet is different. There is one important difference between the two types of inflorescence: character of completion of the main axis (terminal flower is present or absent).
Character of flower open is basipetal. In the Brexia genus forming few-flowered dichasial inflorescences, nested in axils of usual green leaves in upper part of current year shoots. Simple flowers of Parnassia are large, nested on the long generative shoots, going out from rosette, with an amplexicaul leaf. Traces of inflorescence reduction are absent; however, namely such way forming of generative sphere the plant is presented more probable.
In Lepidobotrys staudtii Engl. (Lepidobotryaceae) inflorescences are specific axile (intercalary), fewflowered (more often one-flowered), forming on brachyblasts. Each flower has a small bract at the base.
Many-flowered inflorescences of Stackhousia species nested on the top of generative shoots; there are bracteous botryses or thyrses with gradual transition from usual leafy shoot to bracteous inflorescences. Flowers have a long corolla tube, and sitting on a very short peduncle (almost sitting). In axils of bracts are placed simple dichasia ( S. viminea ), or solitary flowers ( S. monogyna ), with acropetal flower open. Representatives of Celastraceae family and also Stackhousia,
Tripterococcus and Macgregoria genera have 5-merous and 4-cyclous haplostemonous flowers, in compared with
5-merous and 5-cyclous obdiplostemonous flowers of
Brexia and Parnassia . Tubular flowers of Stackhousia and Tripterococcus (but not Macgregoria ) arise by nonperfect postgenital fusion of petals. It is very rare among
Angiospermae. For Celastraceae species we may observe, apparently, late congenital fusion of floral elements (perianth, stamens, sometimes – base of gynoecium), as result of the process, mature flower have nectariferous disk, for Parnassia is fused of laid stamen primordia at the expense of the common base growth, which results in a ('pseudo-')nectaries staminodes of mature flower. In flowers of Brexia fusion is absent, and nectaries-staminodes in mature flower consist of peripheral part of androecium, which appeared as threadlike appendages on the late stages of floral development with strongly reduced vasculature. So, these taxa demonstrate initial stages of the zygomorphy forming, which affected the perianth only (for Stackhousia and
Tripterococcus – androecium also, because it has 3 long and 2 short stamens).
Diversity and evolution of inflorescences in
1
Malpighiales
Prenner, G 1
Royal Botanic Gardens, Kew, UK
The rosid eudicot order Malpighiales, which was drastically recircumscribed following molecular phylogenetic analyses, now comprises more than 16,000 species in c. 700 genera and 37families. Despite a series of molecular phylogenetic analyses, some deep-node relationships within the order remain poorly resolved.
The order encompasses tremendous diversity in morphology and ecology, including some morphological oddballs such as submerged thalloid aquatics
(Podostemaceae), holoparasites (Rafflesiaceae) and leafless cactus-like succulents (Euphorbiaceae). Thus, a reinvestigation of morphological data in the order is

appropriate, both for improved understanding of possible evolutionary pathways and production of revised and refined comparative data that will ultimately help to better resolve relationships. Here I will present an overview of the diversity of inflorescence morphology in
Malpighiales and discuss selected cases such as
Euphorbiaceae in more detail. Floral structures in the order range from the largest flowers among angiosperms in Rafflesia through amentiferous wind-pollinated flowers in the temperate Salicaceae (including Populus ), to highly reduced flowers aggregated into flowerlike pseudanthia in Euphorbia . The entire range of basic inflorescence types (raceme, cyme, thyrse and panicle) occurs in the order, as well as special cases of inflorescence architecture that defy assignment to particular types. This diversity makes the order
Malpighiales a potentially useful subject for a study of inflorescence evolution.
Phylogenetic perspectives on the assembly of high elevation floras in the Neotropics
Donoghue, M 1
1 Dept of Ecology and Evolutionary Biology, Yale
University, USA
How were plants assembled into the higher elevation habitats created through mountain building in the
Neotropics? How were patterns of assembly influenced by changes in the degree of isolation of North and South
America? I review evidence from phylogenetic studies that relates to these questions, focusing especially on the apparent movement of plant groups into South America from the north, and their subsequent rapid radiation at high elevations in the Andes. These considerations bear on the more general question of the likelihood of in situ evolution into new habitats versus the movement of preadapted plant lineages into these habitats.
Recent assembly of the savannas of central Brazil
Simon, MF 1 , Batalha, MA 2
1
2
Embrapa Recursos Genéticos e Biotecnologia, Brazil;
Universidade Federal de São Carlos, Brazil
Modern savannas are thought to have evolved within the last 8 million years, coinciding with the rise to dominance of flammable C4 grasses and the consequent expansion of fire-prone grass dominated ecosystems worldwide. Recent studies focused on the Cerrado in
South America, the world’s most species-rich tropical savanna, have provided insights on the origins and historical assembly of the savanna biome. Timecalibrated plant phylogenies suggest that Cerrado lineages started to diversify less than 10 million years ago, a time frame compatible with the hypothesis of recent (late Miocene) evolution of modern fire adapted savanna vegetation. The growing number of plant phylogenies shows evidence of recently derived Cerrado clades, and multiple independent evolution of fireadapted lineages. Cerrado lineages are strongly associated with adaptations to withstand fire and have sister groups in largely fire-free, nearby wet forest, seasonally dry forest, subtropical grassland, or wetland vegetation. These findings imply that the Cerrado formed in situ via recent and frequent adaptive shifts to resist fire, rather than via dispersal of lineages already adapted to fire. The location of the Cerrado surrounded by a diverse array of species-rich biomes, and the apparently modest adaptive barrier posed by fire, are likely to have contributed to high levels of species richness. Analyses of the phylogenetic structure of biological communities have been used to answer questions regarding contemporary ecological forces that assemble local biotas. Cerrado plant communities show phylogenetic overdispersion when compared to other fire-free vegetation types, suggesting that fire in this case is not a major environmental filter to the ingress of plant lineages. The absence of phylogenetic clustering in the
Cerrado is mainly due to the persistence of long-lived resprouting species from different groups of plants.
Evidence from population genetics studies of fireadapted and fire-sensitive congeneric species show low genetic divergence between savanna and wet forest species pairs, suggesting easy of biome shift through the convergent evolution of fire related traits such as thick corky bark and fire-adapted life forms. Adaptive shifts between ecological zones can play an important role in the generation of species diversity. Our results show that fire adaptation and Cerrado occurrence are phylogenetically labile with multiple independent lineages in disparate plant families. Frequent adaptive shifts and phylogenetic overdispersion in Cerrado plant communities suggest that fire does not pose a significant adaptive barrier to shifts between biomes. Conversely, the boundaries appear to have been porous to the recruitment of lineages from a range of fire-free vegetation types. The assembly of the endemic rich
Cerrado flora via frequent recent adaptive shifts to resist fire stands in contrast to the widespread support for ideas that lineages tend to maintain their ancestral ecologies
(phylogenetic niche conservatism). The emerging picture of Cerrado origins based on dated phylogenies, phylogenetic community structure and population genetics, is of recent diversification of endemic plant lineages, driven by the common trigger of fire adaptation and facilitated by ease of adaptation across plant groups from diverse surrounding biomes.
Origin and evolution of plant diversity in the
Brazilian Atlantic Forest: the Gesneriaceae radiation as case study
Perret, M 1 , Chautems, A 1 , Guignard, M 2 , Salamin, N 2
1 Conservatory and Botanical Garden of Geneva,
Switzerland; 2 Dept of Ecology and Evolution, University of Lausanne, Switzerland
The Brazilian Atlantic forest is known for its striking level of species diversity and endemism, as well as for being one of the most threatened tropical forest in the world. However, the biogeographical history of this hotspot of biodiversity and the mechanisms of speciation that have contributed to the present-day patterns of species richness are largely unknown. To gain insight
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into these issues, we explore the temporal and biogeographic context of diversification in the Brazilian
Atlantic forest using the Gesneriaceae as a model group.
This plant family is distributed in various biomes of tropical America, including the Brazilian Atlantic forest that comprises 105 endemics out of a total of 127 species
(83%). A time-calibrated phylogenetic tree of the
Neotropical Gesneriaceae was produced based on a combined analysis of six molecular markers and a sample representing all the major Neotropical lineages and 90% of the species occurring the Atlantic forest. This phylogenetic tree indicates that Gesneriaceae diversity in the Brazilian Atlantic forest mainly results from in situ radiation of two independent lineages – the rock outcrop plants Sinningieae (90 spp.) and the epiphytic
Codonanthe-Nematanthus (45 spp.). Both clades started to diversify during the Oligocene, a period coinciding with the onset of aridity on the Brazilian Plateau that caused the contraction of the Atlantic and Amazone rain forests. Reconstruction of the ancestral areas in these clades indicates a marked disjunction between northern and southern portions of the Atlantic forest and frequent biotic exchanges between coastal rain forests and their neighboring inland areas. To further explore the evolutionary process that gave rise to this endemic
Atlantic forest flora, we analysed patterns of range overlaps and ecological niche changes among sister clades. The relative importance of geographical isolation and ecological shifts in these speciation events will be discussed.
Finding the needle in the haystack: does introgression play a role in speciation in Central American
Begonia
?
Twyford, AD 1,2 , Harrison, N 1 , Ennos, RA 3 , Kidner, CA 1
1 Royal Botanic Garden Edinburgh, UK; 2 Institute of
Molecular Plant Sciences, University of Edinburgh, UK;
3 Institute of Evolutionary Biology, University of
Edinburgh, UK
Understanding the patterns and processes underlying the formation of species-rich tropical biomes is a major goal of plant phylogeographic and biogeographic studies. The use of spatial genetic data both at the population and species level has shed light on the tempo and mode of speciation in these biomes, and increased our understanding of the patterns of speciation. However, the relative importance of hybridisation and introgression in contributing new lineages to tropical biomes has largely been overlooked. We are investigating these processes in
Central American Begonia , a morphologically diverse group of 650 species which has rapidly speciated since the Early Miocene. We present data from both nuclear and chloroplast microsatellite markers derived from transcriptome and whole chloroplast sequences, in order to investigate population dynamics in the group. An emphasis is placed on two widespread weedy Begonia species from Mexico ( B. heracleifolia and B. nelumbiifolia) , and we discuss both the broad spatial genetic patterns within these species, as well as local patterns in Begonia hybrid swarms. Comparison between nuclear microsatellites and chloroplast haplotype data is used to assess different signatures of pollen and seed dispersal. Genetic data is then placed in the context of greenhouse crosses and morphological examination of
210 the hybrid swarms. Little population genetic work has been done on widespread tropical herbaceous plants, and understanding spatial genetic structure will aid our understanding of speciation in this taxonomically complex genus, as well as the potential genetic reservoir to acclimate to a rapidly changing climate.
Using new genes and techniques to untangle the evolution of problematic taxa: an example from
1
Lantaneae (Verbenaceae)
Lu-Irving, P 1 , Olmstead, RG 1
University of Washington, Seattle, USA
The remarkable floristic diversity of the Neotropics is characterized by numerous species-rich, largely endemic lineages – many of which have turbulent taxonomic histories. Complex patterns of morphological parallelism and intermediacy have obscured natural relationships among members of these lineages; classification schemes at the tribal, generic and species levels are traditionally problematic, and molecular data reveal them to be unnatural. With an increasing range of modern tools available to the international systematic community, great progress has been made in the last several years in untangling evolutionary history within problematic neotropical taxa. Here we present as an example our work in Lantaneae: a morphologically diverse group of some 400 named species, representing the largest radiation within Verbenaceae. Most members occur in the Neotropics, where they form a widespread and important element of the flora; some species are native to
Africa, and the Lantana camara complex has spread across the globe as noxious weeds. As in many other neotropical groups, a contrast between dry fruits and fleshy, animal-dispersed fruits exists in Lantaneae – it is the traditional basis upon which the two major genera,
Lippia and Lantana, are separated. However, substantial morphological intermediacy blurs the generic boundary, and raises the question of what pattern the evolution of the different fruit types followed. With growing interest in sorting out the taxonomy of this difficult group, and growing recognition of the serious worldwide ecological and economic impacts of Lantana camara, there is a clear need for a well-resolved phylogenetic hypothesis for Lantaneae. We took a multi-locus approach to address this need, resolving the pattern of species diversification among a broad representative sample that covers the morphological, taxonomic and geographic diversity of this group. Our results highlight the effectiveness of the newly-developed PPR gene family as phylogenetic tools, and reveal that none of the major genera of Lantaneae are monophyletic as currently circumscribed: fleshy fruits have evolved multiple times independently from dry-fruited ancestors; Lantana camara and its relatives form a lineage distant from that which contains the majority of Lantana species sampled.
The phylogeny allows deeper questions about the evolution of geographic range together with fruit type in
Lantaneae to be explored, and can be used as groundwork in broader studies of general evolutionary patterns across multiple neotropical lineages.

Phylogeny and biogeography in Solanaceae,
Verbenaceae, and Bignoniaceae: a comparison of continental and intercontinental diversification patterns
Olmstead, R 1
1 University of Washington, Seattle, USA
Recent molecular phylogenetic studies of Solanales and
Lamiales show that Solanaceae, Verbenaceae, and
Bignoniaceae all diversified in South America. Estimated dates for the stem lineages of all three clades imply origins in late Cretaceous, at which time the present continents had separated from the united Gondwanan continent. In each clade, a few lineages are represented in
Africa, Asia, and North America. In most cases, the position of lineages of Old World plants on their respective phylogenies suggests much younger origins for these lineages, implying long-distance dispersal as the primary mechanism leading to contemporary distributions. Detailed phylogenetic hypotheses for these clades permit explicit enumeration of the number of events leading to the extant distributions. In contrast to this limited intercontinental distribution, a broad distribution across almost all major lineages within each family suggests little in the way of geographic constraints on intracontinental diversification. Resolution among major clades of Lamiales makes it difficult to establish sister groups for Bignoniaceae and
Verbenaceae, but a well-resolved phylogenetic hypothesis for Solanales implies that the origin of
Solanaceae may have predated the breakup of Gondwana and that extinction may be responsible for the near absence of Old World lineages among the extant early diverging lineages of Solanaceae
Botanical evolution in the Neotropics: a palaeontological perspective
Jaramillo, C 1
1 Smithsonian Tropical Research Institute, Panama
The consequences of global warming on tropical vegetation are unknown. We can use empirical examples in earth history to understand the behavior of tropical biotas during past climate change. During the onset of the
Paleocene Eocene Thermal Maximum (PETM, 55.5
Million years ago) worldwide temperature increased by
6-8°C in ~10,000 years. Temperatures in tropical regions reached 32-33°C, rising at a rate of 0.05°C per century.
This is similar to the temperature rise during the 20th century (0.6 °C). Modern ecological studies suggest that high temperatures could be deleterious to tropical rainforest plants. We studied the pollen record of several sites in Colombia and Venezuela to understand the effects of the PETM on the vegetation. The pollen record suggests a major floristic radiation, with many new species and families arising rapidly, at the onset of the warming during the earliest Eocene. This radiation seems to correlate with the radiation of polypod ferns and orchids that has been seen in phylogenetic studies.
Uppermost Paleocene palynofloras (56-58 My) have a low diversity and are widespread across the region, while lowermost Eocene palynofloras (56-55 My) have a higher diversity and are more restricted geographically with a higher degree of change across the landscape.
Plants extinction levels did not increased during the onset of the PETM, while there was a sharp increase in diversification rates. Results suggest the PETM warming had a major and long-lasting impact on the history of the neotropical rainforests.
1
Lowland tropical rain forests in the Neotropics
Antonelli, A 1 , Pennington, RT 2
Gothenburg Botanical Garden, Sweden; 2 Tropical
Diversity Section, Royal Botanic Garden Edinburgh, UK
Lowland tropical rain forests occur throughout the neotropical region and are characterised by very high levels of plant species richness. There is growing evidence that this seemingly homogeneous ecosystem has been formed by different processes in different areas, and that different taxa in the same region have diversified in response to different evolutionary processes. These processes may include or relate to: soil adaptations; biotic interactions with pollinators, dispersers and herbivores; niche conservatism; dispersal ability; the effect of rainfall, temperature and area; mountain formation; hydrological changes. Although no overarching theory is likely to explain the historical assembly of species richness in the Neotropics in general and lowland rain forests in particular, future studies could aim at quantifying the relative contribution of potential influences on diversification. We argue that well-sampled, confidently dated molecular phylogenies of key plant groups constitute one of the most powerful tools for linking these different fields of science and discerning between competing hypotheses of diversification.
1
How well do we understand the biogeography of
Neotropical plants?
Christenhusz, M 1
Botanical Garden and Herbarium, Finnish Museum of
Natural History, Finland
The relationships of some Neotropical plant groups can be puzzling. Some well-known families, such as
Bromeliaceae, Cactaceae and Caricaceae, all have one to a few species in Africa, and these do not seem puzzling because they clearly represent a recent long-distance dispersal – these groups are of recent origin, so longdistance dispersal is assumed. Other examples are yet older, but the relationship is clear so we view these as indicating relict distributions, as is found in
Strelitziaceae, with Phenacospermum in the Guianas,
Strelitzia in South Africa and Ravenala in Madagascar.
Yet other families do not clearly seem to have distributions indicating that long-distance dispersal has been frequent, but the advent of molecular systematics has shown this to be otherwise. The families exhibiting this pattern include Fabaceae and Lauraceae. These are old families in which one might have expected plate tectonics to play a major role, but this appears not to be the case. In yet other old families, such as Marattiaceae and Orchidaceae, we find the expected pattern suggesting that tectonics is the primary factor, with clades clearly restricted to one tropical region. In these families there
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are genera (Marattiaceae) or tribes/subtribes
(Orchidaceae) largely to exclusively found on one continent. These patterns appear to have little to do with seed/spore dispersability and will require more research to determine why they have occurred. Many additional clades that no one anticipated also fit into the relictual category, as in Strelitziaceae (above), and molecular studies are discovering more of these, the most recent of which is a new, previously unrecognized order
Huerteales, to which we have recently demonstrated that a newly recognized family, Petenaeaceae (based on
Petenaea , described in 1962, as Elaeocarpaceae), belongs. Petenaeaceae was found to be related to the recently described South African family Gerrardinaceae.
The distribution pattern of a Central American species distantly related to an African genus follows the pattern in other families of Huerteales, for instance, in
Tapisciaceae, Tapiscia is restricted to Central China, whereas the related genus Huertea is restricted to
Neotropical mountains. Overall, there are many more connections between the Neotropics and Old World tropics, both Asia and Africa, than previously would have been thought to be the case.
Evolution of Neotropical epiphytes: a lesson from
Bromelioideae (Bromeliaceae)
Silvestro, D 1,2,4 , Schulte, K 3
Benko-Iseppon, AM 6 ,
, Heller, S
Zizka, G 3
3 , Leme, EMC 5 ,
1 Botany and Molecular Evolution, Research Institute
Senckenberg, Germany;
Research Centre;
2 Biodiversity and Climate
3 Australian Tropical Herbarium,
James Cook University, Australia; 4 Inst. Ecology,
Evolution and Diversity, Faculty Biosciences, Goethe-
University, Germany; 5 Herbarium Bradeanum, Brazil;
6 Depto. De Genética, Universidade Federal De
Pernambuco, UFPE/CCB, Brazil
A specific feature of the Neotropics is the extraordinary high species diversity of epiphytic flowering plants.
Besides families with worldwide distribution like
Orchidaceae and Araceae, a number of families with exclusively or predominantly Neotropical distribution contributes considerably to that diversity. Among the latter, Bromeliaceae are the most successful group with over 1,100 epiphytic species. We investigated the evolution and radiation of Bromelioideae as a model case for the conquest of the epiphytic habitat in the
Neotropics. Within the monophyletic subfamily
Bromelioideae (33 genera />800 species, most of them epiphytic), terrestrial basal groups invaded the epiphytic habitat followed by extensive radiation there, especially in the Atlantic Rain Forest. This process was combined with a spread from Andean habitats to Eastern South
America. In addition to succulence and water absorbing trichomes, the key innovations of tank habit and crassulacean acid metabolism (CAM) were essential for the evolutionary success. Our phylogenetic reconstruction is based on nuclear (PRK, phyC) and plastid markers ( mat K, atp Brbc L, trn K, trn L, trn L trn F), and dated applying a relaxed molecular clock to reconstruct divergence times of the major lineages. While the majority of the species is the result of a comparatively recent radiation (core bromelioids:
Aechmea and related genera), major evolutionary changes are concentrated at the base of the subfamily
(terrestrial–epiphytic, C3–CAM, succulent – tank habit;
212 basal bromelioids and eu-bromelioids. Nuclear data provide conflicting phylogenetic signals, especially within the core bromelioids. These conflicts are best explained by a complex history of hybridization and introgression. We will discuss the importance of these phenomena for the diversification of the group especially in relation to biogeography, past climatic changes and anthropogenic habitat destruction. Distribution data and ecological niche modelling allow the reconstruction of niche evolution along the phylogeny. The results are presented and discussed in the light of timeframe, as well as paleoclimate and geomorphological events in South
America.
Species diversity, biogeography and conservation of
Neotropical dry forests
Pennington, RT 1
Lavin, M 5
, Särkinen, T 2 , Hughes, C 3 , Dexter, K 4 ,
2
1 Royal Botanic Garden Edinburgh, Edinburgh, UK;
Botany Dept, Natural History Museum, London, UK;
3 Institute of Systematic Botany, University of Zürich,
Switzerland;
Biologique, CNRS, Université Paul Sabatier, Toulouse,
France; 5
4 Laboratoire Evolution et Diversité
Plant Sciences and Plant Pathology, Montana
State University, Bozeman, USA
Neotropical seasonally dry forests show a high beta diversity, caused by few woody species being widespread between disjunct areas of this vegetation, and many of these areas have high levels of endemism. Isolated fragments of SDTF contain geologically old monophyletic radiations of endemic plant species, and intraspecific genetic variation is highly geographically structured. These patterns indicate a stable, dispersal limited SDTF system. SDTF species tend to belong to larger clades confined to this vegetation, suggesting phylogenetic niche conservatism. Phylogenetic niche conservatism between SDTF and neighbouring vegetations such as woody savannas is confirmed by a phylogenetic community approach that considers the entire flora, and not just exemplar clades. We suggest that SDTF represents a distinctive theatre of evolution
(metacommunity or biome) for woody plants.
Phylogenetic and biogeographic patterns differ in woody plants from tropical rain forests and savannas, suggesting a hypothesis that broad ecological settings strongly influence plant diversification in the tropics.
Can phylogenetic signal, character displacement or random phenotypic drift explain the morphological variation in the genus
Geonoma
(Arecaceae)?
1
Roncal, J 4 , Henderson, A 1
Sodre Cardoso, S 1
, Borchsenius, F
, Balslev, H 1
1 , Ricardo
Institut De Recherche Pour Le Developpement (IRD)
Plant clades may exhibit little or wide morphological variation as a result of 1) the retention of ancestral characteristics or phylogenetic signal, 2) character displacement, or 3) random phenotypic drift. Geonoma is one of the largest and most variable genera of American palms and its taxonomy is controversial. To assess which evolutionary scenario could explain the morphological variation in Geonoma (Arecaceae), we performed a

Mantel test between phylogenetic and morphological distances of 54 Geonoma species, and tested for phylogenetic signal using Blomberg’s K-statistic. To obtain a patristic distance matrix for Geonoma , we constructed a molecular phylogeny of tribe Geonoma teae using three nuclear DNA regions: PRK, RPB2 and
CISP4, and sampling 61.2% of species in the tribe. The phylogeny placed Pholidostachys as sister to the remaining genera. Asterogyne was sister to a clade consisting of Calyptronoma and Calyptrogyne , two main clades were resolved within Geonoma , and we found evidence of intragenomic polymorphism. A positive relationship between the patristic and a 26-discretecharacter distance matrix (R2=0.55, p<0.001) supported the phylogenetic signal scenario. No relationship was evident using a 17-quantitative-variable distance matrix
(R2=0.07, p=0.13), and K-values were all close to 0, suggesting random phenotypic drift. If most morphological variables traditionally used to classify
Geonoma evolved randomly, their little selective value might explain Geonoma 's challenging taxonomy.
1
Molecular phylogenetics of
Acacia
Miller, J 1 , Murphy, D 2
Center for Australian National Biodiversity Research,
Canberra, Australia; 2 Royal Botanic Gardens
Melbourne, Australia
The genus Acacia is the largest plant genus in Australia and is a key component of many ecosystems, especially in arid regions. We present molecular phylogenetic results from plastid and nuclear DNA regions for over
500 species. These data, calibrated with multiple fossils, record the divergence dates of the major lineages with the major expansion of the genus correlating with aridification of the continent. Vegetatively, Acacia is a diverse genus with some species maintaining bipinnate leaves while most species develop phyllodes. Our results trace the multiple changes in leaf architecture and their correlation with environmental zones. Inflorescence characters are less diverse than vegetative characters in
Acacia . The species vary in inflorescence shape, either globose or spicate, and in some species the inflorescence can be arranged in racemes. The evolution of these inflorescence characters is more conserved with these characters defining large monophyletic assemblages of
Acacia . These data resolve major lineages of Acacia that have not been previously reported. We will present the phylogenetic data and present a foundation for a new informal classification of Acacia .
Plant–insect co-diversification as an indicator of community evolution
McLeish, M 1
1 University of Stellenbosch, South Africa
Plants and their insect associates represent an enormous constituent of terrestrial species and represent a particularly versatile system for explaining geographical and phylogenetic history at the community level. The evolutionary diversification of host Acacia and phytophagous gall-thrips that specialise on Acacia likely respond to changing ecological and environmental conditions in alternative but non-independent processes.
Determining the magnitude of differences in these processes should be partially explained by addressing long-standing co-evolution, co-speciation, and sequential speciation hypotheses. Alternatively, phylogenetically conservative traits apparent in contemporary interactions do not necessarily have to concur with historical associations where ecological fitting might operate.
Reconciling of phylogenetic congruence between host and parasite is used to investigate the co-diversification of Australian Acacia (Phyllodineae) and gall-inducing thrips (Thysanoptera). Diversification rate signatures of host and parasite are compared to determine the most appropriate co-diversification hypothesis(es). Unique and shared attributes of galler and host Acacia diversification are discussed in context with the evolution of Australian biomes.
Seasonal partitioning of shared pollinators in
1 southern Australian acacias
Stone, G 1 , Prescott, M 1 , Amos, N 2 , Burd, M 1
Institute of Evolutionary Biology, School of Biological
Sciences, Edinburgh University, UK; 2 Ecology and
Conservation Biology, School of Biological Sciences,
Monash University, Australia
The legume genus Acacia (Leguminosae: Mimosoideae) is one of the most important groups of woody plants in
Australasia. The pollination ecology of entire assemblages of closely related taxa (acacias in the genera
Vachellia and Senegalia ) has only been studied in depth in seasonally arid environments in Kenya, Tanzania and
Mexico (Raine 2001). Most studies of Australian Acacia communities have so far been restricted to species living in the extreme temperatures and low species richness zones of arid western and central Australia. In seasonally arid habitats in Mexico and eastern Africa, Vachellia and
Senegalia acacias commonly flower together in space and time (coflower) following seasonal rainfall. Flowers of coflowering acacias are also often visited by the same insects, a situation that can lead to competition for pollination among coflowering species. Where interacting plant species have similar floral morphology
(as acacias do), shared pollinators can be partitioned in time. Tanzanian and Mexican acacias that coflower cannot separate in seasonal time instead show evidence of temporal partitioning on a daily timescale, driven by between-species differences in the timing of pollen release. Here we examine the extent to which similar patterns are seen in Acacia assemblages in temperate southern Australia. We analysed the seasonal flowering behaviour of an assemblage of seven Acacia species (A. acinacea, A. dealbata, A. pycnantha, A. melanoxylon, A. paradoxa, A. genistifolia and A. mearnsii ) growing in a temperate habitat near Melbourne, Victoria. Our expectation is that lower seasonality in rainfall should allow greater potential for seasonal partitioning of shared pollinators than Mexico and East Africa. We ask the
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following questions: (1) Do southern Australian acacias share pollinators? (2) What are the seasonal flowering patterns for each Acacia species? (3) To what extent is the distribution of species flowering peaks through the season compatible with the predictions of competitive displacement? We use literature data extracted from a
Victoria-wide floral database to discuss the generality of the patterns we describe for the region, and discuss our results in the light of previous work on acacias sensu lato.
Pollination biology and breeding system of an
Australian
Acacia
in a fragmented agricultural landscape
Henery, ML 1 , Cunningham, SA 1 , Broadhurst, LM 2
1 CSIRO Ecosystem Sciences, Australia; 2 CSIRO Plant
Industry, Canberra, Australia
The on-going resilience of native vegetation fragments in agricultural landscapes requires that they are sufficiently connected via genetic exchange with other populations, for populations to persist. This is particularly true for outcrossing species, which appears to be the breeding system for many acacias. For the majority of the
Australian flora including Acacia , genetic exchange through pollen movement is facilitated by animals. For such a widespread and dominant component of the
Australian flora we know relatively little about the reproductive biology of the genus. Almost all Australian
Acacia species produce massed displays of small nectarless flowers that are likely to be pollinated by small insects rather than vertebrates. Consequently we would expect distance of pollen dispersal to be short and connectivity of patches of plants to be low, relative to other perennial woody taxa present in a landscape. For an outcrossing Acacia species habitat fragmentation could result in reduction of the quality and supply of suitable pollen, particularly if patches of remnant vegetation contain small numbers of closely related individuals.
Acacias can be positively or negatively affected by disturbances such as fire and grazing and the in many landscapes these are now strongly driven by human intervention. We investigated the breeding system, pollinator interactions and population genetics of Acacia dealbata to determine what combined effect these have on gene flow and population dynamics across its distribution in a heavily modified landscape. We also discuss the pollination biology of Acacia and how reproductive strategies in the genus have evolved.
Microbial diversity and genetic identity as drivers of productivity in interactions between
Acacia
and nitrogen-fixing rhizobia
Barrett, LG 1 , Broadhurst, LM 1 , Thrall, PH 1
1 CSIRO Plant Industry, Canberra, Australia
Plants in the genus Acacia can receive substantial benefits from forming symbiotic associations with nitrogen-fixing rhizobial bacteria. It is widely recognised that such mutualisms are generally important in driving plant productivity in both natural and agricultural ecosystems. However, we still lack a broad understanding of how genetic identity and diversity of
214 soil symbionts relate to plant performance and ecological function. Improved knowledge of links between the genetic structure of mutualistic networks and ecological function will provide insights into basic ecological and evolutionary drivers of ecosystem productivity, together with applied value in terms of understanding dynamics of community assembly for ecosystem restoration of degraded landscapes. In this talk we will summarise results from recent field and glasshouse studies of
Acacia –rhizobia interactions investigating how spatial variation in rhizobial community structure and symbiotic effectiveness relate to host species identity and environmental factors. We will also present results from experimental work investigating causal links between species identity, genetic diversity and symbiotic function.
Climatic rarity and its possible implications on the distribution of highly endemic species of
Acacia
in
Australia
Gonzalez-Orozco, CE 1 , Miller, J 1 , Knerr, N 1
1 CSIRO, Centre for Australian National Biodiversity
Research, Canberra, Australia
Bio-geographers face the challenge of explaining why certain regions contain a high number of endemic species. Climatic variables such as rainfall and temperature may play a key role in establishing high levels of endemism. Due to the increasing instability of the climate, there is a need to investigate the relationship between endemic species and their local climate. Recent spatial mapping tools and high-resolution continental climatic databases provide a unique opportunity to combine both the distribution range of endemic species with different climatic variables. Here we use the genus
Acacia in Australia to explore whether endemic species with a small geographic range are distributed in regions with unique climates. The first step involves identifying and mapping the localities with the highest scores of endemism at a grid cell level. Then, a climatic characterisation of the highly endemic regions is required. Finally, a statistical correlation analysis is conducted between the climatic and the highest scores of endemism at the grid cell level. Our research thus far has identified the main centres of endemism and species richness at 0.25º (approximately 25 km2) grid cell resolution for the Australian Acacia and we are currently working on the climatic analysis. Our research identifies the climatic variables that define the restricted habitats that local or regional endemic species prefer and consequently the climatic effects on the distribution of endemic species of Acacia .
Deep-node relationships in Apiales: the role of taxon and character sampling
Plunkett, GM 1 , Nicolas, AN 2 , Lowry II, PP 3

1 Cullman Program for Molecular Systematics, New York
Botanical Garden, Bronx, USA; 2 Institute of Systematic
Botany, New York Botanical Garden, USA; 3 Missouri
Botanical Garden, St Louis, USA, and Département
Systématique & Évolution, Muséum National d'Histoire
Naturelle, Paris, France
The dicot order Apiales has been plagued by epic difficulties in identifying 'natural' groups and in understanding evolutionary relationships among these groups. Past efforts have focused most intensively on questions at the rank of species and genus, but virtually every taxonomic level has suffered from these problems, from the circumscription of species to the delimitation of families. Phylogenetic systematics, particularly when incorporating molecular characters, has provided a completely new framework for understanding evolutionary relationships in Apiales, but continued progress depends on integrating two important lessons, still frequently ignored: the need for intensive taxon sampling (both of ingroups and outgroups) and the importance of using characters derived from a diversity of sources. We present an overview of results from the past 15 years (1994–2009) together with new phylogenetic hypotheses of relationships within the order based on multiple plastid markers to highlight key aspects of these lessons and to provide new insights. At the deepest levels, for example, broad sampling of taxa from throughout the eudicots provided early indications that Pittosporaceae belonged to Apiales, as did
Torricelliaceae and Griselinaceae (but not Cornalean elements such as Diplopanax and Helwingia).
Subsequent molecular studies have strengthened support for these early findings, as have additional data derived from structural characters (e.g., cytology, embryology, floral anatomy and development). By contrast, the placement of Pittosporaceae in relation to other groups of
Apiales has been highly unstable in studies using only one or two molecular markers, but this has been resolved in recent studies with vastly improved character sampling. Similarly, some studies suggested an Apialean placement of Pennantia , but this result rested on very limited taxon sampling and has found almost no support from structural data. Realignment of several other groups in Apiales would almost certainly not have been possible without broad sampling (of both taxa and characters).
Notable examples include the segregation of
Myodocarpus and Delarbrea from Araliaceae (as
Myodocarpaceae), the dissolution of Apiaceae subfamily
Hydrocotyloideae (into seven different lineages), the recognition of two new subfamilies of Apiaceae
(Mackinlayoideae and Azorelloideae), the paraphyly of the apiaceous subfamily Apiaceae in relation to
Saniculoideae (leading to the 'proto-apioid' concept), the polyphyly of Polyscias in relation to six other araliad genera, and the gross polyphyly of Schefflera
(Araliaceae). Molecular data, however, must also be carefully scrutinized, particularly to identify problems resulting from polyploidy and paralogy (as in the
Melanesian Schefflera clade). To summarize our current state of knowledge, we present and compare several new phylogenetic hypotheses based on multiple molecular data sets. Results from these studies include the placement of Pennantia, resolution of the early diversification of Araliaceae, the placement of several enigmatic araliad lineages, the non-monophyly of the majority of azorelloid genera, and relationships among the major (subfamilial) lineages of Apiaceae.
Sexual systems in Apiales: why is andromonoecy so common?
Schlessman, M 1
1 Vassar College, New York, USA
Apiales exhibit a variety of sexual systems, including dioecy, hermaphroditism, and andromonoecy. Of these, andromonoecy, i.e. all plant bearing both perfect
(bisexual) and staminate (male) flowers, is unusually common. The high incidence of andromonoecy in
Apiales is noteworthy given the rarity of this sexual system in angiosperms as a whole. Why are so many species of Apiales andromonoecious? Here I examine the origins of andromonoecy in Apiales and the reasons for its prevalence in the order. From a theoretical perspective, andromonoecy is best viewed in the context of variation in sex allocation among the flowers produced by individual plants. Sequential blooming, resource depletion, dichogamy, pollinator movements, and selfing rate are among the factors that may affect optimal allocations to pollen and ovules. The extreme rarity of andromonoecy suggests that in order for it to evolve and persist, either one of these factors must have a very strong effect, or two or more of them must act in concert.
In Apiales, dichogamy is even more common than andromonoecy. My phylogenetic analysis shows that protandry evolved concomitantly with hermaphroditism in the common ancestor of the four largest families:
Pittosporaceae, Araliaceae, Myodocarpaceae, and
Apiaceae. In Pittosporaceae, the most basal of these families, synchronous interfloral dichogamy is reportedly characteristic of the four hermaphroditic genera (dioecy in Pittosporum is derived). Since flowers are blooming synchronously rather than sequentially, a necessary condition for andromonoecy is absent. Umbellate inflorescences arose in the common ancestor of
Araliaceae + Myodocarpaceae + Apiaceae, the large clade in which all of the andromonoecious Apiales occur.
In Araliaceae, complete synchronous protandry and hermaphroditism are plesiomorphic and common, but andromonoecy has arisen independently a number of times. It appears that in each instance, a transition from synchronous flowering and a single-cycle of protandry to sequential flowering with at least 1.5 cycles of protandry is concomitant with the transition to andromonoecy. The presence or absence of sequential flowering, rather than variation in the morphology of inflorescences, appears to be a primary factor determining whether the sexual system is hermaphroditism or andromonoecy. Araliaceae is an ideal candidate for comparative study of the relative importance of other factors, such as fruit size, as ecological correlates of andromonoecy. In sharp contrast to Araliaceae, andromonoecy is both plesiomorphic and very common in the clade Myodocarpaceae + Apiaceae.
This appears to be the result of at least three factors acting in concert: dichogamy, sequentially blooming orders of umbels, and compound umbels that are compact enough to be a primary functional unit of both interaction with pollinators and sex allocation. Aborting the biovulate ovaries of some flowers within a compound umbel is a quantitative, incremental adjustment within a primary reproductive unit. Although umbels with varying
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proportions of perfect and staminate flowers fit our definition of andromonoecy, they differ in some fundamental ways from the more qualitative somethingor-nothing abortion of multiovulate ovaries of larger flowers that are themselves the primary reproductive units of most andromonecious taxa outside the Apiales.
The systematic significance of bark anatomy in
Apiales
Kotina, E 1 , Nilova, M 1
Van Wyk, BE 1
, Rossouw, A 1 , Oskolskii, A 1
1 Botanical Museum, Komarov Botanical Institute, Russia
,
The bark structure of 17 genera of Araliaceae, two genera of Myodocarpaceae, eight genera of Apiaceae, three genera of Pittosporaceae and one genus each of
Griseliniaceae and Pennantiaceae were examined. All the members of Apiaceae, Araliaceae, Myodocarpaceae and
Pittosporaceae studied share the presence of secretory canals in the cortex and secondary phloem, the presence of two types of axial parenchyma cells in secondary phloem (sheaths surrounding the secretory canals and groups accompanying sieve tubes) and the absence of fibres in the secondary phloem. These features can be regarded as synapomorphic for the suborder Apiineae
Plunkett & Lowry, the core group of Apiales. In the two basal groups of the order Apiales (Griseliniaceae and
Pennantiaceae), secretory canals are absent. The presence of calcium oxalate crystals in the primary cortex and secondary phloem indicates a close relationship among members of the order but the types of crystals and their location are very useful characters for distinguishing some families and genera. For example, the occurrence of styloids in the parenchyma cells of the
Myodocarpaceae points to a relationship between this family and Pittosporaceae. The occurrence of prismatic crystals in cells of the axial parenchyma and in the rays probably represents the ancestral condition in Araliaceae.
Prismatic crystals in the cortical parenchyma cells of the
Mackinlayoideae indicate a close relation with
Myodocarpaceae and Pittosporaceae and confirm the isolated position of the Apiaceae. The presence and type of crystals and sclereids in periderm cells and the presence of secretory canal in the rays confirm the isolated position of the type section of Schefflera and, therefore, the polyphyly of this genus. Schefflera selloi is remarkable in having schizogenous secretory cavities in the phellem, an uncommon feature not yet reported from any other Araliaceae or related taxa. Homogeneous secondary phloem rays are typical for the
Myodocarpaceae but they are rarely also present in other groups. The Mackinlayoideae can be distinguished by the thickened tangential walls of their phellem cells. Most members of the Asian Palmate group of Araliaceae share phellem cells with thickened walls, the occurrence of druses and sclereids in the axial parenchyma. All
Apiaceae clades as well as Pittosporaceae are characterized by a radial type of dilatation in the secondary phloem, whereas among the Araliaceae this type of dilatation has been noted only for some basal clades and Polyscias sensu lato. In Griseliniaceae and
Pennantiaceae, as well as in the main group of
Araliaceae, dilated secondary phloem are diffuse-radial.
The sieve tube members in most Apiaceae (except
Mackinlayoideae) and Pittosporaceae are less than 0.6
216 mm long, whereas in most Araliaceae they are 0.5 to 0.8 mm long and in Myodocarpaceae they exceed 0.8 mm.
The length of sieve tube members shows no clear trend in its evolution within the Araliaceae and related families.
Therefore most bark features are of diagnostic importance in at least some genera or species groups.
Phylogenetic significance of African Apiaceae and a new tribal classification for the Protoapioids
Magee, AR 1, 2 , Van Wyk, B-E 2 , Tilney, PM 2 , Downie,
1
SR 3
Compton Herbarium, South African National
Biodiversity Institute, Cape Town, South Africa; 2Dept of Botany and Plant Biotechnology, University of
3
2
Johannesburg, Johannesburg, South Africa; Dept of
Plant Biology, University of Illinois at Urbana-
Champaign, Urbana, USA
A renewed global research effort is currently underway to produce a modern classification system for the
Apiaceae to replace the outdated and largely unnatural treatment proposed by Drude (1897–98) more than a hundred years ago. As the comparatively small sub-
Saharan African contingent had, until relatively recently, remained largely unstudied, little provision was made for them in Drude’s classification. The high incidence of woodiness and unusual leaf and particularly fruit anatomical characters, as well as the many isolated and anomalous taxa, indicate that the African genera are critical to an understanding of higher order relationships within the family. Affinities amongst these hitherto poorly known sub-Saharan African genera will be explored using morphological, anatomical and molecular sequence data. In particular, much focus will be on the early diverging lineages within the Apioideae–
Saniculoideae clade through phylogenetic analyses of the cpDNA trn Qtrn K 5’exon region. Using this dataset, the evolutionary history of characters traditionally used to define the subfamilies Apioideae and Saniculoideae are explored by ancestral trait reconstruction. Our analyses demonstrate that the subfamily Apioideae as traditionally circumscribed is non-monophyletic when the African taxa are considered and the characters traditionally used to segregate it from its sister subfamily Saniculoideae are reconstructed to be either plesiomorphies or synapomorphies restricted to the upper lineages of the subfamily. A broader morphologically congruent circumscription is therefore proposed which includes the
Saniculoideae rather as the tribe Saniculeae within the early lineages of the Apioideae s.l. In this broadened circumscription, the Apioideae are readily distinguished from the other two subfamilies (Azorelloideae and
Mackinlayoideae) by the absence of rhomboidal crystals, the presence of druse crystals scattered throughout the mesocarp (subsequently lost in the upper lineages), the non-woody endocarp and the sporadic presence of true wings. Within the Apioideae s.l., two informal groups are recognised, viz. protoapioids (those early lineages with scattered druse crystals in the fruit mesocarp) and euapioids (the remaining upper lineages where druse crystals, if present, are restricted to the commissure). In order to make provision for the gradient of predominantly African lineages within the protoapioids, eight tribes are recognised of which five are newly described. As a result of this and several other recent

studies, the majority of the African Apioideae have now been placed within some 22 tribes or as yet undescribed major clades, with about a third of the genera within the protoapioid tribes. It is clear that the African taxa represent an important component of the Apiaceae, and substantial rearrangements at almost all infrafamilial levels are required in order to incorporate them into the new emerging classification.
Correlation in fruit and molecular characters in
Eurasian and N African Umbelliferae
Degtjareva, G 1 , Pimenov, M 1 , Vallejo-Roman, C 2
1 Botanical Garden, Biological Faculty, Moscow State
University, Moscow, Russia; 2 AN Belozersky Institute of
Physico-Chemical Biology, Moscow State University,
Russia
In Umbelliferae (Apiaceae), carpological characters traditionally regarded as a basis for taxonomy and diagnostics. At present, the carpological characters of
Umbelliferae of Eurasia and N Africa have been summarized in special carpological descriptive database
(http://umbel.guru.ru), elaborated by Botanical Garden and Research Computing Center of Moscow State
University. On the other hand, during last three decades a molecular classification of Umbelliferae, with special attention to the major Apioideae subfamily, has been elaborated (e.g. Nicolas, Plunkett, 2009; Downie et al.
2010), based on both nrDNA ITS and cpDNA sequence data. This allows us to elucidate correlation between carpological and molecular characters in Umbelliferae.
For the purpose of the analyses, a supertree of the
Umbelliferae based on published and several unpublished molecular phylogenetic trees produced by our group was constructed. The supertree includes more than 200 taxa of Eurasia and N Africa with fruit morphology scored.
Among 50 carpological characters included in the carpological database we selected six ones, being the most widely used in current taxonomic practice in the family: (1) shape of mericarps in transverse section: strongly laterally compressed, slightly laterally compressed, isodiametric or dorsally compressed; (2) ribs of mericarps: primary only or primary and secondary; (3) calyx teeth at fruits: prominent or obsolete; (4) width of mericarp commissure: narrow, medium or wide; (5) vittae: absent, vallecular and commissural, vallecular only, commissural only, cyclic or diffuse; (6) endosperm at commissural face: flat, slightly convex, or slightly concave; with broad, shallow groove; with broad deep groove; with narrow, deep groove; mushroom-shaped. The distributions of fruit characters in molecular tree are very complex. Taxa with secondary ribs (trait (2)) are strongly concentrated in two neighbour clades ( Torilis Clade and Daucus Clade), with two exclusions only, Artedia and Exoacantha , which need further investigation. In trait (4) stronger correlation is observed only in some tree fragments – taxa with broad commissure are concentrated is such molecular clades as Heracleum Clade and Ferula Clade, and the mixture of genera with broad and middle-sized commissure form almost exclusively a big clade, containing Karatavia, Tommasinia, Dichoropetalum and some other genera. On the contrary, the trait (3) demonstrates rather stochastic distribution, although there are some molecular clades with well developed calyx teeth almost exclusively (mainly in basal part of tree) or without prominent teeth ( Elaeosticta Clade,
Rumia – Ridolfia Clade, Scandix Clade). Mericarp compession in dorsal-ventral direction (trait (1)) traditionally used to discriminate 'peucedanoid' and
'apioid' genera in Apioideae. The taxa with strongly compressed mericarps form several molecular clades, other variants are scattered in tree. The trait (6) used in
Umbelliferae taxonomy since A.P.Decandolle, shows moderate correlation with molecular clades. In trait (5), several small molecular clades are composed by taxa with cyclic vittae; in general, correlation is low. In general, obtained distribution patterns of carpological characters in molecular tree supports the traditional view on key role of fruit specialization on evolution of
Umbelliferae and are of interest for analysis at the level of closely related genera or within polymorphic genera.
The position of a Madagascan genus
Kaliphora
in the system of angiosperms: insights from the standpoint of carpology
Konstantinova, AI 1 , Yembaturova, EY 2
1 MV Lomonosov State University, Moscow; 2 Russian
State Agrarian University, KA Timiryazev MSAA, Russia
Previously a monotypic genus Kaliphora was placed in
Hydrangeales (Takhtajan 1997), which presently no longer exists (APG III; Takhtajan 2009). Takhtajan &
Trifonova (1999) pointed out that amongst Hydrangeales, the taxa most similar to Kaliphora in drupaceous fruit structure were Melanophyllaceae and Corokiaceae, while in seed structure Montiniaceae had the greatest degree of resemblance. Now Montiniaceae, Corokiaceae and
Melanophyllaceae (Torricelliaceae sensu Plunkett et al.
2004) are placed in different orders (APG III; Takhtajan
2009) – Solanales, Asterales and Apiales respectively.
Kaliphora madagascariensis Hook.f. is a relatively small, well-branched tree with simple leaves from central
Madagascar. Kaliphora ’s indehiscent dimeric semiinferior fruit possesses a lot of archaic traits, such as: peculiar decurrent stigmata, no secretoty ducts in mature fruits, a thick but completely discontinuous stone, a
'protocolumn' formed from the ventral bundles. The closely related taxa from former Hydrangeales –
Montinia and Corokia – do not show any similarity to
Kaliphora in fruit structure. In Montinia , an inferior syncarpous bicarpellate dehiscent capsule develops
(Yembaturova 2001). Thin filmy septum holds multiple imbricate winged seeds. Corokia is known to have a fleshy inferior pyrenarium (normally two-loculed but one-seeded, rarely the second seed is present). The fleshy portion of the pericarp is found to possess some peculiar traits (Yembaturova, 2001). Nothing similar has ever been reported in Kaliphora , so apparently fruit morphology and anatomy does not support a close affinity of Kaliphora , Montinia and Corokia . Torricellia and Melanophylla , currently placed in Apiales (Plunkett et al., 2004; APG III) as part of the family
Torricelliaceae, possess fleshy inferior 2- to 4-loculed, yet one-seeded pyrenaria. The similarity between drupaceous fruits of Kaliphoraceae and Torricelliaceae is also very approximate. Kaliphora fruits differ markedly from those of Torricellia and Melanophylla since their stones are not only dimeric, but also discrete. In
Torricellia and Melanophylla pyrenes are fused and there
217

are three to four locules within a single stone, as well as in a few other Cornales sensu Takhtajan 1997
(Yembaturova et al., 2009). Although morphological similarities between Kaliphora ceae and Torricelliaceae are not very significant, we consider both of them part of
Apiales. Kaliphora fruit structure shows hydrocotyloid pattern: a dimeric fruit with two discrete lignified pyrenes which has no apioid- or saniculoid-type secretory system at maturity. Out of all related genera studied, Kaliphora is most similar to Mackinlaya – a shrub with palmately compound or simple leaves from
Australasian and Malesian regions. Apiopetalum -
Mackinlaya group is a well-supported basally branched clade within Apiales (Lowry et al., 2001). Characters shared by Kaliphora and Mackinlaya are: fleshy dimeric fruits with peculiar decurrent stigmata and nondifferentiated surface ultrasculpture, more than five vascular bundles and unpronounced non-winged ribs, thick stones, discontinuous near the commissure at the dehiscence area, a 'protocolumn' incapable of separating.
All of the above support possible position of Kaliphora within or close to Apiaceae-Mackinlayoideae group.
Apparently, Kaliphora and Mackinlaya possess ancestral character states and represent an early lineage in Apiales.
Out of Australia: divergence and biogeographic history in Apiales
Nicolas, AN 1 , Plunkett, GM 2
1 Institute of Systematic Botany, New York Botanical
2 Garden, Bronx, USA; Cullman Program for Molecular
Systematics, New York Botanical Garden, USA
Evidence from extant distributions and the fossil record suggests that Apiales have occupied all major phytogeographic regions. To date, published estimates for the age of Apiales have varied widely but have been based on very few representatives from the order. The two most speciose families, Araliaceae and Apiaceae, are broadly distributed across the tropical and temperate regions (respectively), whereas the remaining families
(Myodocarpaceae, Pittosporaceae, Griseliniaceae,
Torriceliaceae, and Pennantiaceae) have much more limited distributions. Traditional theories indicated an origin of Araliaceae in the paleotropics during the
Cretaceous and the derivation of Apiaceae from protoaraliaceous stock. To test such theories, we made use of paleobotanical evidence and gene phylogenies to estimate the age of Apiales and the times of divergence of its major clades, and to reconstruct the biogeographic history of the order. We used sequence data from various plastid markers (including rpl16 intron, trn Dtrn Ytrn Etrn T region, and rpl 32trn L spacer), and from the mitochondrial (nad1 intron 5) and nuclear (duplicate copies of rpb2) genomes. Conclusions were drawn from a sampling of more than 300 taxa, representing all major clades across Apiales. We estimated times of divergence using a relaxed molecular clock with lognormal distribution, implemented in the BEAST package, together with six calibration points, including three macrofossils (fruit from Torriceliaceae and Araliaceae, and leaves from Araliaceae) and three pollen microfossils from Apiaceae. We used three different approaches to biogeographic reconstructions, Lagrange, S-DIVA, and
DIVA. In Lagrange, we tested different scenarios with stratified and non-stratified models of range
218 relationships, and accounted for phylogenetic uncertainty by running the analyses with alternative placements of unresolved clades. Results suggest an origin of Apiales approximately 110 Ma and that all families originated during the Cretaceous, more than 80 Ma. Biogeographic reconstructions show an origin in Australasia for all families. Within that region, Australia is the most likely center of origin, but New Zealand and New Caledonia seem to have played major roles in preserving early lineages (e.g., Griseliniaceae and Myodocarpaceae) and in the diversification of many clades in Araliaceae and
Pittosporaceae. The most-recently diverging family,
Apiaceae, also shows an origin in Australia, at least for the family as a whole and two of its early lineages
(subfamily Mackinlayoideae and the Platysace clade), but a South American origin for subfamily
Azorelloideae, and a South African origin for Apioideae
+ Saniculoideae. Vicariance related to the breakup of
Gondwana and the drifting of Madagascar and India explain the divergence of early lineages (e.g.,
Torricelliaceae), but long-distance dispersal played a major role in the more recent diversification events since the late Cretaceous. Major dispersal trends include (1) diversification from Australia and New Zealand through
Antarctica and subantarctic islands into southern South
America (e.g., Azorelloideae, Hydrocotyle clade of
Araliaceae, Griselinia ) and in some cases into the high
Andes (e.g., Azorella ); (2) dispersal through Australasia into Asia and Europe then North and South America
(e.g., Asian-Palmate clade of Araliaceae); (3) dispersal across the Indian Ocean Basin into Madagascar and
Africa (e.g., Araliaceae and Pittosporaceae); and (4) dispersal out of Africa into Europe and Asia (Apioideae).
Historical biogeography of Apiaceae subfamily
Apioideae: from southern Africa to the Northern
1
Hemisphere and back to the south again
Spalik, K 1 , Banasiak, U 1 , Piwczyski, M 2 , Downie, SR 3
University of Warsaw, Warsaw, Poland;
Copernicus University, Toru, Poland;
Illinois at Urbana-Champaign, USA
3
2 Nicolaus
University of
Umbellifer subfamily Apioideae is distributed predominantly in the temperate regions of the Northern
Hemisphere. However, its early branching tribes
Heteromorpheae, Annesorhizeae, Lichtensteinieae,
Choritaenieae, and Marlothielleae occur in southern
Africa, as does the basal clade of the sister subfamily
Saniculoideae, suggesting therefore a southern African origin for both subfamilies. The other subfamilies,
Azorelloideae and Mackinlayoideae, are mostly South
American and Australasian, suggesting a Gondwanan origin for the entire family. With the notable exceptions of Molopospermum and Astydamia , the early lineages of
Apioideae are almost entirely absent from the Northern
Hemisphere. This absence may be recent as evidenced by an early occurrence of Heteromorpha -type and Anginon type pollen in Tertiary sediments in Europe. The appearance of the major branch of Apioideae in the
Northern Hemisphere is dated to the late
Cretaceous/early Tertiary, as is its early divergence into
Mediterranean Bupleurum and Asian core Apioideae. It is the Asian branch of umbellifers that subsequently colonised the entire Northern Hemisphere and diversified into ca. 3000 extant species. The members of this branch

also migrated to the Southern Hemisphere. Estimated using a calibrated molecular phylogeny, the dispersal and diversification of its Australasian members coincided with the mid-Miocene cooling.
Wood diversity within Araliaceae: evolutionary trends or/and geographic patterns
Oskolski, A 1
1 Komarov Botanical Institute, Russia
The pathways of secondary xylem evolution within 38 of the 41 genera of Araliaceae were reconstructed by mapping of the wood character states onto the molecular trees (Plunkett et al. 2004; 2010). The major trends of wood evolution (sensu I. Bailey) cannot be unambiguously distinguished within this family; rather, multiple reversals (the increase of vessel element length, multiplication of bars per scalariform perforation plate, and the transition from simple to scalariform perforation plates) were revealed. In addition, some georgraphic patterns of wood diversity were observed. The evolutionary transition from the scalariform to simple perforation plates occurs only in the Polyscias s. lato clade. This trend is realized as the gradual elimination of vessels or vessel portions within scalariform perforation plates which is not accompanied by a gradual decrease in ithe number of bars per perforation plate. Within
Polyscias , the scalariform perforation plates were found only in two basally divergent subclades with the diversity centers occuring in Malesia and Australia; members of other subclades centered in New Caledonia, Hawaii,
Seychelles, Mascarenes, and Madagascar share exclusively simple perforation plates. The
Tieghemopanax and the ‘New Guinea – Polynesia’ clades are distinctive from the ‘Indian Ocean Basin clade’ by wider rays, and more numerous septate fibres.
Probably, this pattern of wood diversity is associated with early radiation within Polyscias s. lato in the
Australian-Malesian region, and independent diversification of the derived groups in basins of the
Indian and Pacific Oceans. Unlike Polyscias s. lato, the basal subclades within four groups (the Asian Schefflera clade, the nominate section of Schefflera , Meryta , and
Cussonia + Seemannaralia ) are distinctive from the terminal ones by shorter vessel elements, and by mostly or exclusively simple perforation plates. It is worth noting that the members of these basal subclades are widespread in higher latitudes and/or in temperate zones, in contrast to the tropical or subtropical distribution of their terminal subclades. The occurrence of advanced character states in basal subclades is more likely the effect of specialization of the secondary xylem during expansion of the taxa through difficult environments, rather than an ancestral condition. Within the Araliaceae, therefore, the most specialized wood features (sensu I.W.
Bailey) occur in the species near the edges of the geographic ranges of their subclades, and occur irrespectively of their phylogenetic position. As this pattern suggests, the Baileyan major trends may be considered as adaptive tendencies associated with the expansion of new plant taxa from their tropical origins into drier and/or cooler habitats. This suggestion needs further consideration for other monophyletic plant groups besides the Araliaceae family.
Molecular evolutionary history of early branching angiosperms
Samain, M-S
Neinhuis, C 2
1 , Salomo, K 2
, Wanke, S 2
, Smith, JF 3 , Feild, T 4 ,
4
1 Ghent University, Belgium;
Dresden, Germany; 3
Monash University, Australia
2 Technische Universität
Boise State University, Idaho, USA;
The earliest divergent lineages of angiosperms include representatives of many of the various growth forms, habits, pollination syndromes, and physiologies that appear in more recently evolved lineages. Understanding the evolution and origin of these traits among the first lineages of extant angiosperms can provide insights into the evolution and origins of these traits among monocots and eudicots. However, to fully understand the evolutionary processes, we must first understand the evolutionary patterns, and the timing of when divergent events occurred. Phylogenetic analyses using DNA sequence data have recently resolved relationships among most major angiosperm lineages with only a few ambiguously placed groups remaining to be resolved.
These phylogenetic estimates have been valuable toward understanding the evolution of a wide array of morphological traits among the eudicots, but there have been few investigations to date that have studied the evolution and diversification among the earliest divergent lineages of angiosperms. Likewise, molecular dating has been applied across numerous angiosperm lineages, but has not been used extensively among the earliest divergent lineages of angiosperms themselves. The exceptions often use early divergent lineages to place the date of the origin of flowering plants, but have not examined the timing of evolutionary events among these lineages themselves. Despite what is often considered a fossil-rich group of species, many lineages of early angiosperms, such as Piperales, have few to no fossils, and where the fossils exist they are often more recent than the age of the lineage is presumed to be. Here, we use both chloroplast ( trn Kmat Kpsb A) and low copy nuclear (phytochrome A) DNA regions to resolve relationships among lineages of the earliest divergent flowering plants. We also aim to provide a means of molecular dating among these lineages. To accomplish this goal we have sampled extant taxa to include clades that can be dated with a minimum age based on fossils with well-established dates, which can be confidently placed on the tree. Our sampling, based on 300+ species representing more than 100 genera, allows for multiple independent calibration points. The results of our analyses allow us to put dates on numerous lineages that lack adequate fossil representatives without resorting to secondary calibration methods.
Success and failure of growth forms diversification in
1 basal angiosperms
Isnard, S 1
TU- Dresden, Institut für Botanik, Germany
219

The order Piperales is the most species-rich clade among basal angiosperms (referred here as lineages branching before the core of monocot/eudicot), comprising more than 4000 species. The existence of virtually all growth and life forms, such as geophytes, epiphytes, aquatics, parasites, annuals, perennials, herbs, succulents, lianas, shrubs and trees, is symptomatic for the success of this clade in many ecosystems. As such, this early diverging lineage underwent tremendous growth and life form diversification with all their adaptations. In that sense,
Piperales constitute a model for diversification of life in angiosperm at the edge of basal angiosperm and monocot/eudicot radiation. This remarkable diversification has occurred dissymmetrical in the lineage. Some genera belong to the top 10 angiosperm genera with respect to species richness ( Piper and
Peperomia – in total ~3600 species, with a pan-tropical distribution) while other genera are species poor with restricted distributions ( Verhuellia , Manekia and Zippelia
– 9 species in total). This 'dissymmetry' in success of evolution provides a framework to address the question on 'key' traits (adaptations that push radiation) versus evolutionary 'dead-ends'. After reviewing growth form diversity and related anatomical and architectural features in early Angiosperms ( Amborella ,
Austrobaileyales), we will focus on comparative patterns of growth form diversification in Piperales, tracing the anatomical, biomechanical and architectural adaptations behind. In doing so we discuss how some wood characteristics and architectural features appear to have inbuilt mechanical and developmental constraints which may have profound effects on the subsequent evolution of growth forms in the different clades. We will propose key traits that may have been responsible for species radiation in these major clades and highlight character or combination of characters which may shed light onto the
'bottle neck' observed in species poor genera.
Limited floral resources and pollination systems in basal angiosperms
Thien, L 1 , Fan, J-H 2 , Chen, Z-D 2 , Devall, M 3
1 Tulane University, Louisiana, USA;
3
2 Chinese Academy of Sciences, Beijing, China; United States Dept of
Agriculture Forest Service, Center for Bottomland
Research, USA
Limited Floral Resources and Pollination Systems in
Basal Angiosperms The first three branches of the angiosperm phylogenetic tree are composed of plants pollinated by insects and/or wind. Floral resources for visiting insects, however, are limited and include: pollen, nectar, starch bodies, heat, and brood sites: pollen being the main food for visiting insects. In some of these basal flowering plants, nectar is produced in small amounts on perianth parts usually at the base of tepals, but in
Schisandraceae also by stamens. Nectaries on carpels and extra-floral parts (e.g. leaves) evolved only in the monocots and eudicots millions of years later. Plants in the first three branches have dry stigmas and do not produce 'pollination droplets' as found in gymnosperms; in Nymphaeales stigmas may be wet but do not function as a food source. With limited floral resources, extant species evolved various combinations of food, floral color, and morphological features (e.g. false stamens) to deceive and attract insects. As a result, a single or
220 combination of resources can determine the pollinator and evolution of a pollination system. Some pollination systems have endured for long periods of time, e.g., fossil flowers of water lilies (80–100 mya) have the same structure as extant species and were probably pollinated by beetles; studies of large flowered water lilies in Africa indicate long term pollination by beetles. In
Schisandraceae (third branch) Kadsura and Schisandra have unisexual flowers and Illicium bisexual flowers.
Pollination systems in members of Schisandraceae are primarily based on flies and beetles, however, pollen is the main food source in species of Schisandra in China and in relictual areas gall midges have co-opted the original pollination system. On the other hand in
Illicium, heat and brood sites are the main resources of the plants, in which post-sexual heat production and secretions of fluid by stamens feed the larvae. These gall midge pollination systems (1–2 pollinators per plant species) occur in the Sino-Japanese Flora in many different plant communities. In North America,
Schisandra and Illicium pollination systems are generalist but specialist in the Sino-Japanese Flora. In
Schisandra it is hypothesized that since the gall midges mate on abandoned spider webs located near the plants, that pollen was initially exploited to enrich eggs, and species of gall midges could increase reproduction by feeding on a dependable food source. Subsequently, because the gall midges are efficient pollinators, the life cycle of the plants and insects evolved into a tight association (but not co-evolution) in old stable plant communities in the Sino-Japanese Flora. The differences between the liquid-feeding Clinodiplosis gall midge pollination system in some species of Illicium and the female pollen-eating gall midge Resseliella pollination system in Schisandra appear to be fostered by different floral resources in the two lineages. The age of these pollination systems is not known, but given the rarity of these pollination systems and its occurrence in old forest communities, it may be a remnant of a once more widespread pollination system.
1
Female gametophyte development and competition in early-diverging lineages of flowering plants
Bachelier, JB 1 , Friedman, WE
University of Colorado, USA
1
Just over the last twenty years, molecular studies have greatly contributed to the resolution of phylogenetic relationships amongst flowering plants at all systematic levels. In addition, the resolution of a basal grade of early-diverging lineages provided a new and robust framework for ancestral character reconstruction, which revealed that the first flowering plants were surprisingly far more diverse than it was long assumed. Here, we report in Trimenia , an extant member of the
Austrobaileyales, a remarkable mechanism of female gametophyte development. In each ovule, multiple female gametophytes are differentiated remotely from the site of fertilization and they initiate a pattern of intercellular tip growth in the nucellus central tissue.
They grow concurrently like tubes, intertwined with each other from the base to the apex of the nucellus, and only one of them will yield the single embryo of each seed.
Such a development strikingly resembles that of pollen tubes growing in the transmitting tissue of the carpel, and

suggests that Trimenia has evolved a unique mechanism of female gametophyte competition. However, our phylogenetic analysis reveals that within ovule female gametophyte competition is by no means unique among angiosperms. While the initiation of multiple female gametophytes exhibiting a pattern of tip growth is unusual and only occurs in a few unrelated taxa, the occasional initiation of more than one female gametophyte is frequent, particularly among extant members of basal clades of flowering plants. Our analyses suggest that a mechanism of female gametophyte (egg) competition, in addition to male gametophyte (sperm) competition and maternal mate choice, may have characterized the first flowering plants and have contributed to their rapid diversification and tremendous diversity.
Evidence of diversification, hybridization and adaptation in seven Indian representatives of the genus
Nymphaea
based on morphology, RAPD, PCR-
RFLP and sequence data of the nrDNA ITS region, chloroplast trn
K intron, mat
K and rbc
L gene
Dkhar, J 1 , Kumaria, S 1 , Tandon, P 1
1 Plant Biotechnology Laboratory, Centre for Advanced
Studies in Botany, North Eastern Hill University, India
An investigation of diversification, hybridization, and adaptation in seven Indian representatives of the genus
Nymphaea, viz. N. alba var. rubra , N. caerulea , N . × marliacea , N. nouchali , N. pubescens , N. rubra and N. tetragona was conducted using morphology, RAPD,
PCR-RFLP and sequence data of the ITS region, chloroplast trn K intron, mat K and rbc L gene.
Morphological examinations revealed considerable variation among the species investigated. Phenotypic features with multiple character states (more than two), which may be useful for identification, were recorded for rhizome shape, leaf margin, petal number and color, and pollen shape. An interesting observation was made for two sympatric specimens of N. nouchali , viz. N. nouchali
JD 06 and N. nouchali JD 07 resembling each other in all aspects but differing only in flower color. The change in flower color could be due to the presence of fungal endophyte, identified as Choanephora conjuncta , isolated from the leaves of N. nouchali JD 07 .
Furthermore, the white colored petals of N. pubescens exhibited a tinge of pink at the apex, resembling the petals color of N. rubra . Barring N. caerulea and N. nouchali , RAPD analysis of randomly selected individuals from each species showed low genetic variation at both levels (intra and inter populations). The variability detected for N. caerulea may be due to gene flow (Nm=0.0737) from another coexisting species ( N. alba var. rubra ) or through cross-pollination sustained by the high rate of seed formation (100%). In N. nouchali , considerable morphological variation accounts for the high genetic variability (68.09%). PCR-RFLP of the ITS region revealed additional fragments, exceeding the expected size of the ITS region, in N. alba var. rubra , N.
× marliacea , N. rubra and N. pubescens . Sequencing of the ITS region indicated that the extra fragments, manifested as additional signals in the sequencing chromatogram, in N. pubescens may be due to random mutations occurring in some of the ITS paralogues.
However, recent hybridization and introgression may be the reason for the additional signals depicted in the chromatograms of N. alba var. rubra and N. rubra . This suggestion is supported by the exact nucleotide sequence matches of the chloroplast trn K intron, mat K and rbc L gene with their respective putative maternal parents.
Interestingly, mat K gene of N. tetragona , a rare and endangered plant of India, revealed relatively higher number of non-synonymous substitutions as compared to synonymous substitutions. Molecular evolutionary analysis indicated that three of these sites may be under mild selective pressures. Such adaptive changes at the
DNA and protein sequence level of mat K gene may have been associated with the colonization of N. tetragona , suggesting that it could have migrated from China. By integrating both molecular and non-molecular characters, phylogeny of each species investigated based on neighbor joining, maximum parsimony, maximum likelihood and Bayesian inference is presented and discussed.
What can we learn from the genomics of basal angiosperms?
1
DePamphilis, C 1
Dept of Biology, Penn State University, University Park,
USA
Although the genomes of more than two dozen plant species have now been sequenced and assembled, most of the species chosen for complete sequencing have been restricted to economically important plants that fall in a few major lineages. Basal angiosperms include only a few thousand species, but these diverse plants represent the earliest surviving branches from the angiosperm 'tree of life'. Large EST datasets from several basal angiosperms are providing a detailed view of the genes present in early angiosperms and how these gene sets differed from those of gymnosperms and the well characterized grass and eudicot lineages. They also provide a rich source of information for phylogenetic and molecular evolutionary studies. When these data are used in phylogenomic studies of thousands of gene families, gene duplication patterns reveal large peaks in the frequency of gene duplication that occurred shortly before the radiations of seed plants and angiosperms, suggesting that both groups are likely to have been ancestrally polyploid. I will present an overview of deep coverage sequencing and assembly of the genome of
Amborella trichopoda , the sister species to all other flowering plants. A reference sequence of Amborella will provide an effective outgroup to enable comparative genomics across the angiosperms, and enable detailed estimation of the ancestral genomic content, gene order, and ancestral gene families of flowering plants.
The
Euphorbia
Planetary Biodiversity Inventory
(PBI) project – a global inventory and phylogenetic study of the spurges
Berry, P 1
221

1 University of Michigan, USA
Euphorbia , a monophyletic lineage of about 2,300 species, is a tremendously diverse genus of worldwide distribution. Morphologically, it can be characterized by the presence of a pseudanthial cyathium. Physiologically, it is the only genus known to include C
3
, C
4
and CAM photosynthetic systems. Through the U.S. National
Science Foundation's Planetary Biodiversity Inventory programme, we have been conducting studies on the phylogeny, taxonomy, and other aspects of the adaptive radiation of this giant genus. This symposium presents results to date of this 5-year collective effort, including a robust molecular backbone of the infrageneric phylogeny, new explorations and floristic updates, evidence for multiple evolution and convergence of succulent habits and photosynthetic systems, and webbased tools to facilitate dissemination of information on
Euphorbia . The complex biogeography of the genus will be a focal point of the symposium. This introductory talk will provide an overview of the field work, laboratory work, and results of our collaborative efforts to date.
The giant genus
Euphorbia
(Euphorbiaceae): deep phylogenetics and major evolutionary patterns
Wurdack, KJ 1 , Horn, JW 1 , van Ee, BW 2
1 Smithsonian Institution, Dept of Botany; 2 Black Hills
State University, South Dakota, USA
Euphorbia , with ~2300 recognized species, is the second largest genus of angiosperms. Unlike many other clades of 'giant genera,' structural, ecological, and physiological diversity in Euphorbia is on par with species richness. In particular, growth form diversity in Euphorbia is probably more varied than in any other genus and it is the only land plant genus expressing all three major photosynthetic pathways (C
3
, C
4
, CAM). To understand the evolution of such biologically important traits and to assess the importance of the origin of novel phenotypes on differential diversification in Euphorbia , we developed a robustly resolved phylogeny for 176 taxa of tribe Euphorbieae (including 161 Euphorbia species) with sequence data from 9 loci spanning all three genomic compartments (~16 kb per taxon) to use as a framework to investigate these questions. The analyses recover four major clades that are successively sister to each other, beginning with the currently recognized subgenera Esula (Clade B of previous studies),
Rhizanthium (Clade A), Euphorbia (Clade C) and
Chamaesyce (Clade D). Character state optimizations reconstruct a woody, non-succulent growth form with a
C3 photosynthetic pathway as ancestral for the genus.
Specialized growth forms and alternative photosynthetic pathways evolved within each of the four subgeneric clades. Two of these four clades (subg. Rhizanthium and
Euphorbia ) have major radiations of cactiform succulents correlated with the origin of obligate CAM photosynthesis; collectively they account for nearly 650 species. There appears to be only a single origin of C
4 photosynthesis in the core clade of species formerly recognized as the genus Chamaesyce , which constitutes a radiation of nearly 300 species. Although a woody, nonsucculent growth form optimizes as the ancestral state along all nodes of the backbone of the phylogeny, the independent evolution of (C
3
) herbs in subgenera Esula
222 and Chamaesyce gave rise to radiations totaling about
600 species.
Euphorbia
subgenus
Rhizanthium
: an entirely Old
World lineage with surprisingly low sequence
1 divergence despite great morphological diversity
Morawetz, JJ 1 , Riina, R 2,3
Rancho Santa Ana Botanic Garden, Claremont, USA;
2 Real Jardín Botánico, Madrid, Spain;
Michigan, Ann Arbor, USA
3 University of
Here we present the first detailed hypothesis of relationships within the exclusively Old World subgenus
Rhizanthium based on ITS and ndhF sequence data. With ca. 200 species, subgenus Rhizanthium contains a variety of growth forms, including herbaceous, geophytic, caudiciform, woody, succulent, and semi-succulent woody species. The greatest diversity is achieved among succulent taxa, including the morphologically wellcharacterized 'medusoid' species. Here we support the placement of the woody E. antso , a Madagascan endemic and sole member of subgenus Rhizanthium from the island, sister to the rest of Rhizanthium . The next diverging lineage is the Arabian dwarf caudiciform E. hadramautica . The remaining lineages contain groups of exclusively herbaceous, woody, or succulent species, with the largest clade representing the majority of diversity in succulence found within Rhizanthium .
Despite the presence of morphologically cohesive groups of succulent species, branch lengths in the large succulent clade are surprisingly short, offering no resolution of relationships among these species. Our data support the placement of the spineless pencil-stemmed succulent species E. dregeana and E. larica within subgenus Rhizanthium , with this growth form now present in all four subgenera. These results support the hypothesis of rapid diversification and specialization within arid habitats of the Old World (especially South
Africa), which would result in the short branch lengths recovered for these succulent taxa. Arid radiations are not unique to subgenus Rhizanthium , as the large spineshield group in subgenus Euphorbia shows a similar pattern of diversification in arid habitats.
Euphorbia
subgenus
Esula
: the main temperate radiation of the genus in Eurasia with disjunctions to the New World and Africa
Geltman, D 1 , Peirson, J 2 , Riina, R 2
1 Komarov Botanical Institute, Russian Academy of
Sciences, Russia; 2 University of Michigan, USA
Euphorbia subgenus Esula contains about 470 species that occur mainly in temperate Eurasia, but with smaller, disjunct centers of diversity in the mountains of the Old
World tropics, temperate areas of South Africa, and the
New World. Most of the species are perennial and annual herbs, but shrubs and small trees (which are sometimes succulent) also occur in the clade. The group's characteristic features are annual stems or new growth branches with mostly alternate leaves terminating in a pseudoumbel of branching rays bearing cyathia. The cyathia have 4 or 5 glands, sometimes with horn-like outgrowths or appendages. Boissier in his 1862 treatment

of the genus separated most species of subgenus Esula into section Tithymalus . Prokhanov further developed the classification of the subgenus (1933 and 1964), but amendments to his system have been proposed recently for nontropical Eurasian species. With the aim to build a natural classification of the subgenus and address key evolutionary and biogeographic questions such as the origin and diversification of the New World species, we conducted phylogenetic analyses using two chloroplast regions ( ndh F, trn Ttrn F) and one nuclear region (ITS) with comprehensive, worldwide taxon sampling.
Consistent with results from previous molecular studies, subg. Esula corresponds to Clade B of Euphorbia (one of four main clades of the genus) and is sister to the other three clades (A, D and C). Some succulent species previously placed in the former subgenus Tirucalli , now part of Clade C or subgenus Euphorbia , are nested within
Clade B in our analyses. Our phylogenetic hypothesis shows that subgenus Esula is separated into two major clades, with a third lineage (B-3: E. lathyris ) sister to the other two. Clade B-1 contains ca. 150 species with shiny dark brown seeds, mostly warty fruits, and glands without horns (this includes sections Chamaebuxus,
Helioscopia , and Holophyllum and part of the section
Balsamis ). Clade B-2 consists of the remaining 300+ species characterized by purple seeds, fruits without warts, and glands both with and without horns (this includes all other previously recognized sections). Our results show that annuals, as well as succulents and semisucculents in many cases, have arisen independently, and that recognition of separate sections and subsections based on these characters is not supported. The presence of species from both major clades not only in Eurasia but also in the New World and Africa suggests an early divergence of clades B-1 and B-2. Most New World species fall into one subclade together with some maritime Eurasian species of Mediterranean origin.
Several New World species (e.g., E. purpurea , E. spathulata ) are members of clade B-1, however, suggesting multiple colonizations of subg. Esula from the Old World. Based on our phylogenetic hypothesis and re-examination of morphological characters, a new sectional classification of subg. Esula is being developed.
Euphorbia
subgenus
Euphorbia
: a diverse lineage with regional radiations and repeated evolution of novel growth forms
Dorsey, B 1 , Haevermans, T 2 , Berry, P 1
1
2
University of Michigan, Ann Arbor, Michigan, USA;
Museum National d'Histoire Naturelle, Herbier
National, Paris, France
Recent molecular phylogenetic analyses strongly support the monophyly of the giant genus Euphorbia and its division into four subgenera. Euphorbia subgenus
Euphorbia contains about 700 of the roughly 2300 species in Euphorbia , is arguably the most morphologically diverse and has a pantropical/subtropical distribution. It contains five of the seven formerly segregate genera which have been subsumed into Euphorbia including Cubanthus ,
Pedilanthus , Monadenium , Synadenium , and
Endadenium . In terms of habit this clade contains macrophyllous understory and canopy rainforest trees, green-stemmed, leafless trees and shrubs, leafy shrubs, spiny stem succulents, geophytes, scrambling lianas and herbs. While the subgenus is pantropical, major subclades have radiated regionally most notably in Africa and Madagascar. Here we present a phylogeny of
Euphorbia subgenus Euphorbia based on nuclear ITS and chloroplast matK and ndhF DNA sequence data from a broad sampling of the subgenus. These data resolve the relationships among most major clades that comprise the subgenus with good to excellent support. They also suggest that there have been rapid radiations at several points in the history of this clade. Within this phylogenetic framework we discuss these rapid radiations in both the New and Old Worlds as well as the biogeographic history of the subgenus and multiple derivations of novel growth forms.
C
3
, C
4
and CAM all in one lineage: remarkable physiological diversifications in
Euphorbia
subgenus
Chamaesyce
Yang, Y 1 , Berry, PE
1 University of Michigan, USA
Chamaesyce subgenus Chamaesyce is remarkable in having all three photosynthetic types found in plants: C
C
4
3
,
and CAM. A well-resolved phylogeny is presented including around 250 ingroup species out of the total 610 species in subg. Chamaesyce using nuclear and chloroplast markers. The entire subgenus can be dissected into three parts: 1) a basal Old World grade of
~150 species, suggesting an Old World origin for the subgenus. Within this Old World grade there is a radiation of pencil-stem succulents in South Africa, as well as diverse non-succulent taxa native to warm areas in Africa, Asia and Australia. 2) An Old World-Eastern
Brazil clade nested in the Old World grade, including about 10 species. 3) The bulk of species in subg.
Chamaesyce reside in a clade that is mostly New World and sister to the Old World–Eastern Brazil clade. This
New World clade includes the formerly segregated genera Chamaesyce and Poinsettia , together with the former subgenus Agaloma . Deeply nested within New
World C
3
species in this clade there is a group of four pencil-stem succulents found in subtropical North
America that have CAM photosynthesis. Also in this
New World clade is the formally segregated genus
Chamaesyce (now section Anisophyllum ), which is mostly C
4
. C
4
photosynthesis in this section originated probably at least twice in desert conditions in subtropical
North America, then radiated into around 300 species that are mainly New World. At least eight long-distance dispersal events have occurred during the worldwide expansion of the section, and widespread interspecific hybridization events are inferred by incongruent ITS versus chloroplast markers, together with cloning data from a nuclear low-copy gene region exon 9 of
EMB2765.
223

European botanic gardens working for climate change mitigation and adaptation
Schulman, L 1 , Lehvävirta, S 2
1 Finnish Museum of Natural History, University of
Helsinki, Finland;
Helsinki, Finland
2 Dept of Biosciences, University of
Europe, more than half of the vascular plant flora may become endangered by the year 2080 as a result of climatic changes. In some European cases, successive
Red List evaluations have already documented the first negative developments in the threat status of plants attributable to climate change. The Global Strategy for
Plant Conservation (GSPC) has been adopted under the
Convention on Biological Diversity (CBD) as a response to the plant diversity crisis. Its recently up-dated version for the years 2011–2020 incorporates the challenge presented by changing climates. Botanic gardens of the world, largely through Botanic Gardens Conservation
International (BGCI), have played a central role in the development and implementation of the GSPC, not least in Europe. In June 2009, the Fifth European Botanic
Gardens Congress, EuroGardV, was organised in Finland by the European Consortium of Botanic Gardens, BGCI, and the Helsinki University Botanic Garden, under the title 'Botanic Gardens in the Age of Climate Change'. A total of 127 papers were presented and seven workshops were arranged. More than 200 delegates from 34 countries participated. This prsentation summarises the main outcomes of the congress and draws conclusions thereof. Rapid global change puts demands on the basic functions of botanic gardens in addition to emphasising the need for conservation research and actions. Gardens could provide society with a means of adapting to global change by revamping their traditional role as introduction centres of novel plants, but this requires the gardens to develop, adapt, and continuously follow rigid collection policies and practices. At the same time, botanic gardens should evaluate and re-direct their research activities in order to make a stronger contribution to climate change mitigation and adaptation. In Europe, 42% of regionally threatened species exist in living plant collections and seed banks maintained by botanic gardens. An emerging challenge is that climate change threatens the endurance of the living collections. Furthermore, the first cases of the alteration of the genetic structure of ex situ populations have already been documented. Seed banking requires development as regards the best conditions for the preservation and germination of seeds.
Botanic gardens have the expertise to carry out projects on re-introduction of plants into the wild, which is the ultimate purpose for having ex situ conservation collections, and they have started engaging in this activity also in Europe. The need to apply ex situ approaches much more widely in connection with assisted migration as a response to rapidly shifting climatic regimes is becoming more apparent. Indeed, given this development, botanic gardens with their unique expertise on collecting, storing, propagating and cultivating wild plants are turning into indispensable links in the chain of effective plant conservation actions.
224
Long-term phenology of the conserved dipterocarps
1 and implications for conservation
Zhao, J 1 T , Zhang, Y 1 , Song, F 2
Xishuangbanna Tropical Botanical Garden, Chinese
Academy of Sciences; 2 Yunnan Institute of
Environmental Science, China
Climate change has shown profound impacts on the plants around the globe. In the perspective of plant conservation, how the conserved plants in the botanical garden responded to the ongoing climate change is a critical question needed to be concerned. Phenology is considered to be a reliable indicator of the climate change. In tropical areas, studies with a phenological data set of more than ten years are still few. We dealt with the phenological records (1974–1998) of 5 Dipterocarpaceae species in Xishuangbanna Tropical Botanical Garden
(XTBG), southwestern China to answer: (1) How did the phenology of the 5 species vary since 1970s? (2) Did the variation of phenology relate to the regional climate change in Xishuangbanna? If so, what were the key affecting factors? (3) Did the conserved dipterocarps benefit from the regional climate change or not? Linear regression and quadratic polynomial regression was used to reveal the inter-annual trends of four phenological events (budburst, growing season, flowering and flowering period). Multiple regression models between phenological data and climatic factors (mean temperature, rainfall and sunshine accumulation) were performed to seek the driving factors for each event. The results show that the four events presented nonlinear trends for most species during the studied period, and a general turning-point in mid-1980s was displayed, which is about 5 years later than the turning point of climate change. Generally, the date of budburst and flowering has delayed by 1.08 and 0.41 days per year, respectively.
Most of these phenological variations can attribute to the regional climate change in Xishuangbanna. Still, the budburst date began to advance since mid-1980s, which was mainly induced by the rising temperature. Moreover, early events in an annual cycle are more affected by temperature, while, the late ones are more affected by rainfall and sunshine duration. The prolonged growing season (1.48 days per year) can promote the stem diameter increment of the plants and further benefit the biomass accumulation. The shortened flowering period
(1.42 days per year) may be a threat to the plant pollination; somehow, it can be different among species depending on the flowering time and the dynamic of their pollinators' population.
Monitoring shifts of genetic diversity in natural populations of
Capsella bursa-pastoris
– intraspecies gene pool dynamics under the influence of climate change
Bernhardt, K-G 1 , Kropf, M 1 , Hameister, S 1 , Neuffer, B 1
1 University of Natural Resources and Life Sciences,
Vienna, and Dept of Integrative Biology and Biodiversity
Research Institute of Botany and Botanical Garden,
Austria
Consequences of climatic fluctuations due to the general warming are ever-present in Austria, for instance on

agricultural land and on pioneer sites. Such consequences include changes in edaphic conditions like increasing drought and drift. Additionally, the global change also effects a shift in the vegetation period which enables the natural invasion of plant species from other floristic regions. In particular, weeds and pioneer species from the sub-Mediterranean and Mediterranean region are common invaders on agricultural land and pioneer sites in Austria. In these habitats Crambe hispanica ,
Diplotaxis erucoides and Capsella rubella are just a few recent examples for the current immigration of (sub)-
Mediterranean taxa to Austria. Demographic processes and especially gene pool dynamics under the influence of climatic fluctuations will be monitored in natural plant populations. For this purpose, the study species should be characterised by a weedy ecology enabling a rapid response to environmental shifts without a dramatic loss in fitness, demanding high phenotypic plasticity and genetic variability. The annual to biennial Capsella bursa-pastoris was selected as a model species, since its ecotypic differentiation, germination behaviour and fitness are well known and also the colonising and longdistance dispersal abilities have been studied intensively.
Currently, two Botanical Gardens have established longterm sites to investigate demographic parameters (lifecycle, seed bank) for Capsella bursa-pastoris . Additional
Botanical Gardens have announced their cooperation and on each site, seeds of at least 20 individuals will be sampled annually over a period of ten years. Fingerprint and isozyme analyses may then reveal changes in genetic diversity patterns over time. The results may also reflect a successful colonization of a pre-adapted genotype, i.e. a possible invasion of warmth-dependent ecotypes/genotypes of Capsella bursa-pastoris . In this context it will be highly interesting to correlate genetic results with ecological factors.
1
Ways of reducing the climate footprint of a botanic garden
Löhne, C 1 , Schomaker, K 1 , Stevens, A-D 1
Botanischer Garten Und Botanisches Museum Berlin-
Dahlem, Germany
The effects of environmental and climate change on biological diversity and on humanity are manifold.
Changing climatic conditions and changing habitats force many species to the edge of extinction, thereby impairing ecosystem functions and services. Botanic gardens are currently refocusing their efforts in conservation on mitigating the negative effects of climate change on plant diversity. However, even such 'green' institutions as botanic gardens are contributing considerably to CO
2 emission, either directly by heating inefficiently isolated greenhouses or indirectly by using large amounts of peat for cultivating the plant collections. Therefore, botanic gardens need to take on responsibility and introduce new strategies for saving energy and reducing greenhouse gas emissions. Here, we present examples from the Botanic
Garden and Botanical Museum Berlin-Dahlem, where new energy concepts and technological innovations in the greenhouses led to considerable reductions in energy consumption. Furthermore, an ongoing research and development project aims at optimizing recycling of biological matter such as horticultural waste. 'Terra Preta' technology employed in the garden will on the one hand help to recycle waste and thereby sequestrating carbon.
On the other hand 'Terra Preta' will be a valuable and potent soil additive that increases fertility and water retention capacity. The long-term goal of the new energy and recycling concept of the Berlin Botanic Garden is to reach the point of 'zero emission'. This strategy also involves communicating and demonstrating these efforts and the underlying techniques to the broader public.
1
Botanic gardens and their role in plant evolution
Prokhorov, A 1
Petrozavodsk State University, Russia
The activity of botanic gardens has recently been considered with special attention drawn to biodiversity preservation and environmental education. However, botanical gardens would have another mission if seen in terms of remote consequences for the nature and humanbeing. For many centuries botanic gardens have been selecting and cultivating the most sustainable forms of useful plants. Meanwhile the repeated part of collections comprises their major part as each curator strives for picking up the maximum collection, thus, following one of the oldest instincts which ensure survival. Plant introduction is performed by various methods, often several generations of plants passing certain stages of adaptation in intermediate climate conditions are used.
Therefore, a network of reserves of plant genetic resources is formed beyond their natural and climatic habitats. Climatic habitat implies the territory where cultivation of a species in the local temperature and light regimens is possible. In other words, it is a geographic habitat extrapolated to other regions of the world on the basis of its key (hardly variable) climatic parameters. The set of modern information resources concerning biodiversity as well as databases on botanical gardens collections, geo-informational systems gave an opportunity to conduct a large-scale research on mangenerated spread of plants. The change of climatic preferences for a number of Gymnospermae species introduced from Europe, North America and Eastern
Asia is shown on the example of Russian botanic gardens collections. In-situ these plants are characterized by quite a limited habitat which boundaries are defined also by climatic factors. Superposition of Gymnospermae habitats on climatic maps allowed to determine limiting climatic factors for several species even if a model climatic habitat in the Russian territory does not include all the botanical gardens where the plants are being cultivated at the moment. One could suppose that either ecological plasticity or selection of species with the necessary physiological features (and genotypes), or interspecies hybridization which is almost inevitable in botanic gardens especially in case of stepped introduction constitutes the basis for emerging of more sustainable forms. In any case one deals with new taxons which are close to the original species according to their morphological characteristics but differ by high tolerance to certain factors. One can check whether these plants have genetic changes comparing genetic material taken in-situ and from botanic gardens located beyond the climatic habitat of this species. Botanic gardens do not only preserve biodiversity but also enhance it forming the habitat for the human being where he/she is surrounded by familiar plants regardless of the place of living. First
225

and foremost, it is carried out by adaptation of economically important taxons to new environmental and climatic conditions. One should not regard the nature as something permanent and requiring preservation in its unchangeable species composition. It is impossible.
However, there is an opportunity to alleviate consequences of negative climate fluctuations including with a help of botanic gardens. Botanic gardens spread throughout the world represent a kind of Noah's arks for plants in case of possible environmental changes.
Strategies for optimizing the conservation value of ex situ living collections of endangered species
Kiehn, M 1
1 Botanical Garden, University of Vienna, Austria
In recent years, living plant collections have gained increasing attention as valuable genetic ressources, especially for rare and endagered species. In order to optimize the impact of such collections for in-situconservation, numerous aspects need to be taken into account, including the consequences of predicted climate change szenarios. Based on experiences obtained in
Austrian Botanic Gardens, problems and obstacles for conservation-oriented living in-situ collections are specified. Strategies for overcoming these negative aspects are proposed - also in the light of potential new challenges arising from climate change.
The importance of New Zealand for understanding the phenomenon of species radiation
Breitwieser, I 1 , Ward, J 2
1
2
Allan Herbarium, Landcare Research, New Zealand;
School of Biological Sciences, University of Canterbury,
New Zealand
In recent years, molecular phylogenetic analyses of the
New Zealand flora have changed our understanding of its diversity and origins. These studies have challenged the traditional view that New Zealand’s biota has been isolated since the breakup of the southern supercontinent,
Gondwana. Under this 'Moa’s Ark' hypothesis, New
Zealand is thought of as the home to relic species undergoing slow changes over long periods of time.
However, we now realize that this view is far too simplistic and a more dynamic, almost tumultuous, view of New Zealand’s biodiversity is emerging.
Contemporary research demonstrates that much of the
New Zealand flora is the result of late Tertiary
(Pliocene–Pleistocene) species radiations. Species radiation has been inferred in numerous plant groups in
New Zealand, raising questions of both evolutionary and conservation interest. Reconstructing the evolutionary history of these radiations is important for understanding the present day distribution and diversity of the flora.
Considering species radiation, what factors have contributed to the diversity present in the New Zealand flora? Key factors in the evolutionary success of a
226 radiation may include innovations in morphology, reproductive features, and physiology, and variation in these traits among closely related species is often related to differences in resource utilization. Abiotic factors are also potential drivers for generating and maintaining floristic diversity, and in New Zealand these include uplift of the Southern Alps, diversity of geological parent materials, Pleistocene glacial cycles, and steep environmental gradients. Studies of the genetic basis of diversification increase our understanding of how evolution on the molecular level has shaped our current biodiversity. Hybridisation and polyploidy have long been thought important for understanding New Zealand’s plant biodiversity. Testing the consequences of these processes requires studies that demonstrate the occurrence of reticulate evolution, and field studies to determine the evolutionary potential of natural hybrids and polyploids. The New Zealand flora provides an ideal system for understanding plant evolutionary and ecological processes as they operate in a more global context. An introduction to recent progress in our understanding of the phenomenon of species radiation in
New Zealand will be presented.
Evolutionary significance of polyploidy in the New
Zealand flora
Murray, B 1
1 The University of Auckland, New Zealand
There is increasing evidence from sequenced plant genomes that whole genome duplication or polyploidy has been a feature of genome evolution in the angiosperms. However, this process is ongoing and more recent polyploid events (neopolyploidy) can be recognized by comparative studies of the morphology, cytogenetics and phylogenetics of polyploids and their diploid relatives. Ancient or paleopolyploids show extensive duplication of syntenic blocks of genes coupled with diploid-like chromosome behaviour and these are recognized by their high basic chromosome number and extensive gene duplication. With chromosome numbers now known for at least one individual from c. 80% of the
New Zealand endemic and indigenous angiosperm flora it is clear that polyploidy, both neo- and paleo-, has been a key element in the evolutionary process. The vast majority of species have basic numbers greater than 7–10 and many of these are paleopolyploids. Neopolyploidy is also widespread and is a feature of many of the most species-rich genera such as Veronica ( Hebe ), Celmisia ,
Ranunculus and Coprosma in the endemic flora. These neopolyploids present interesting questions for analysis.
One involves whether they are autochthonous or whether the initial colonizers were polyploid; both of these scenarios would appear likely with genera such as
Lobelia and Veronica showing clear examples of recent polyploidy though others, like Celmisia , are only represented by polyploids. A second major question is whether the polyploids are autopolyploids that have arisen following genome duplication within species or are allopolyploids, the products of hybridization between species with different genomes followed by polyploidy.
The phylogenetic analysis of many genera suggests the relatively recent arrival of one or a very few ancestors with consequently low levels of sequence divergence between species. This suggests low levels of genome

differentiation and consequent autoploid events.
However, this appears to be contradicted by the presence of diploid-like meiotic pairing in c. 95% of the polyploids studied, which suggests alloploidy. One way to resolve this question is to use molecular cytogenetic techniques coupled with phylogenetic analyses of genera that contain both diploid and polyploid species. This is underway in Plantago , where there are six ploidy levels, and the results show complex patterns of genomic evolution suggesting significant genomic change despite low levels of sequence divergence.
Species delimitation in recent New Zealand species radiations
Meudt, H 1
1 Museum of New Zealand Te Papa Tongarewa, New
Zealand
Plant species radiation is an important process of diversification in New Zealand, a biodiversity hotspot with a unique flora. Even though many New Zealand species may be morphologically and/or ecologically distinct, delimiting the boundaries of species within these radiations and evaluating their conservation status has been difficult for several reasons. For example, many closely-related species are not reproductively isolated, and interspecific hybridization – especially when combined with polyploidy – may create new or intermediate phenotypes. In addition, the phylogenetic relationships of species, as well as the genetic characteristics that distinguish them, are often poorly understood and difficult to establish due to reticulation and low levels of genetic diversity. All these events may obscure species boundaries, and can result in polymorphic and/or paraphyletic species. Although resolving species boundaries in plant species radiations can be challenging, the issue is of particular significance in New Zealand. This is because more than 80% of species are endemic, exhibit highly restricted geographic ranges, and/or are threatened. Taxonomists need to be explicit regarding which operational criteria they use to define species boundaries for their study groups.
Emphasis by different taxonomists on different criteria
(e.g., monophyly at one or multiple DNA loci, morphological diagnosability, ecological distinctiveness) can lead to different conclusions regarding species limits and taxonomy. On the other hand, different types of criteria may be required for delimiting species in different plant groups. Open debate regarding which criteria are most objective and which species concepts are most appropriate is very important. In this talk, I will provide a literature review and synthesis regarding species delimitation in New Zealand plant species radiations, paying particular attention to operational criteria (whether implicit or explicit), difficulties with and resolution of obscure species boundaries, trends over time, and current practice. I will compare and contrast species delimitation of different plant species radiations and discuss interesting trends where possible (e.g., alpine vs. lowland, species radiation vs. non-radiating groups).
In addition, I will focus on specific examples of New
Zealand radiations for which genetic, morphological and ecological data are being generated in diverse and integrative evolutionary studies, including Pachycladon
(Brassicaceae), Craspedia and other Gnaphalieae
(Asteraceae), and Ourisia , Plantago , and Veronica s.l.
(which includes the Hebe complex; Plantaginaceae), among others.
Reconstructing the evolutionary history of species
1 radiations – the impact of hybridisation
Smissen, R 1
Landcare Research, New Zealand
Since the early twentieth century botanists have marvelled at the prominence of wild interspecific hybrids in the New Zealand flora. Cockayne and Allan’s pioneering work documented many putative hybrid combinations and later authors including Dansereau and
Rattenbury mused on the causes and evolutionary importance of hybrids in New Zealand. During the latter years of last century botanical focus shifted away from studies of hybridisation, partly as a result of the prioritisation of economic botany, but the adoption of molecular systematic methods has created resurgent interest in the subject. DNA sequencing and fingerprinting now allow not only robust tests of the parentage of putative hybrids but also allow for the discrimination of first- from later-generation hybrids in wild populations and the potential to measure geneflow between species. Moreover, many authors have attributed phylogenetic incongruence among independent DNA sequence data sets, or between DNA-based and morphology-based phylogenies to the influence of hybridisation, either through introgressive hybridisation
(particularly chloroplast capture) or hybrid speciation.
The impacts of hybridisation on the evolution of New
Zealand species radiations can be observed at all historical stages, from contemporary geneflow between recently diverged species back to ancient hybrid events preceding the radiation of a group in New Zealand, or even predating its arrival. In this talk I will use examples from recent literature (e.g. Ranunculus , Asplenium ,
Pachycladon , everlasting daisies, Ourisia , Pseudopanax ) to illustrate the evolutionary importance of hybridisation in New Zealand plants and examine progress in resolving related problems in phylogenetic reconstruction of species radiations. This will include discussion of the putative hybrid origins of paleopolyploid lineages in
New Zealand, methods for distinguishing between lineage sorting and hybridisation as sources of phylogenetic incongruence, untangling reticulate relationships among species as a result of hybrid speciation, and the estimation of species trees from
AFLP data. While reconstructing the one true species tree is probably an unrealistic and perhaps even inappropriate goal for many species radiations, there is real promise for progress in identifying major clades within species radiations and a more precise understanding of the importance of hybridisation during different phases of species radiation.
Diversification of New Zealand alpine plants: are these radiations adaptive?
Lehnebach, CA 1
1 Museum of New Zealand Te Papa Tongarewa, New
Zealand
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Alpine areas in the North and South Island of New
Zealand harbour a number of plant groups that, despite their recent origin and low genetic diversity, have undergone considerable morphological and ecological diversification. Two of the most outstanding examples of plant species radiation in New Zealand are the genera
Ranunculus and Myosotis , each with over 40 species.
Species within both genera have colonised different habitats, or micro-habitats, within the alpine areas, and exhibit diverse growth habits and leaf and floral characteristics. The extent of phenotypic diversification observed in these two genera has provided the grounds to suggest both radiations are adaptive. This means that diversification of these plant groups has been driven by differential utilisation of resources available in these mountain environments. In general, adaptive radiations of plant, animal and insect groups have been commonly described on Oceanic Islands. So far, whether plant radiations in the New Zealand archipelago are adaptive has not been fully demonstrated. Adaptive radiations can be detected by the presence of four features; rapid speciation, recent common ancestry, phenotypeenvironment correlation and trait utility. Molecular phylogenetic investigations of alpine Ranunculus and preliminary phylogenetic evidence from Myosotis have shown that the New Zealand species of both genera display two of the four features adaptive radiations commonly exhibit: common ancestry (both groups are monophyletic) and rapid speciation (c. 3–5Myr ago for the alpine Ranunculus ). The other two features, namely phenotype-environment correlations and the adaptive significance of the different phenotypic traits, however, have not been fully investigated. In this talk I will focus on the morphological, anatomical and ecological diversification that both plant groups have undergone and, by taking a comparative approach, I will discuss the morphological and anatomical evidence to support phenotype-environment correlations. Specifically, I will address the following questions: How phenotypically diverse are these radiations? Are phenotypically similar species found in similar habitats? Are vegetative characters more diverse than reproductive characters? I will conclude my talk with an introduction to current studies on these genera aiming to understand the physiological and adaptive advantages of such phenotypes.
Origins of the Australian flora – an example from wallaby grasses and allies (Poaceae)
Humphreys, A 1,3 , Pirie, M 2 , Linder, P 1
1 University of Zurich, Switzerland;
Stellenbosch, South Africa;
UK
3
2 University of
Imperial College London,
An integral part of our understanding of large scale processes shaping present day biomes and entire floras comes from the analysis of individual lineages. In the absence of fossil evidence, we provide information on the origins of grass elements of the Australian temperate flora, based on results of molecular phylogenetic and
228 dating analyses. Our focal group, Danthonioideae
(Poaceae), comprises 280 species of C
3
grasses, distributed in temperate regions of the Southern
Hemisphere, North America and Eurasia. There are 42 species native to Australia – some of which were the foundation of the Australian sheep industry – distributed in the southeastern temperate region, with a few species also in the southwestern temperate region, in Tasmania and one species on Lord Howe Island. A phylogeny of
80% of the species, based on cpDNA and nrDNA, indicates at least three colonisations of Australia after the onset of aridification of the Australian mainland: one, probably from Africa, and one from South America, leading to Notochloe microdon and three species of
Plinthanthesis , all of which are geographically and ecologically restricted; one from New Zealand, leading to the restricted alpine species Chionochloa frigida (plus one to Lord Howe Island, C. howensis , not sampled); and one, also from Africa, leading to the widespread austral genus Rytidosperma (wallaby grasses in Australia).
Rytidosperma has crossed the Tasman Sea several times, in both directions and has undergone at least one dispersal to New Guinea and probably two dispersals to
South America, all in the past ~5–10 Ma. Diversification of the 36 species of Rytidosperma native to Australia is therefore likely to have been tightly linked to diversification of the genus as a whole across the
Southern Hemisphere. Due to hybridisation (past and present) between species and multiple ploidy levels within species the interpretation of phylogeny in
Rytidosperma is not straightforward. Nevertheless, we provide an overview of possible ecological aspects of the diversification of wallaby grasses and allies in Australia and in the Southern Hemisphere.
Origin and evolution of the Australian Cucurbitaceae
Schaefer, H 1 , Sebastian, P 2 , Telford, IH 3 , Renner, SS 2
1 Harvard University, USA; 2
Germany; 3
University of Munich, LMU,
University of New England, Armidale,
Australia
The gourd family, Cucurbitaceae, comprises almost 1000 species of mainly tropical and subtropical climbers and creepers. Today’s flora of Australia is surprisingly poor in Cucurbitaceae: Only twelve genera and about 40 species are indigenous to the Australian continent
( Austrobryonia, Benincasa, Cucumis, Diplocyclos, Luffa,
Momordica, Muellerargia, Neoalsomitra,
Nothoalsomitra, Sicyos, Trichosanthes , and Zehneria ).
This low Australian species diversity is in marked contrast with the minimally 20 independent dispersal events into Australia based on molecular phylogenies and ancestral area reconstructions. The largest Australian radiations are in the genera Austrobryonia , Cucumis , and
Trichosanthes and comprise only three to four species each. Two genera are endemic to Australia: the monotypic Nothoalsomitra , a liana of Southeastern
Queensland's Eucalyptus and rain forests, and
Austrobryonia with four species of dry clay soil habitats in Central and Western Australia. Austrobryonia is remarkable because of its close relationship to a
Mediterranean and southern Eurasian clade ( Bryonia and
Ecballium ). This is unique among the native Australian cucurbits, which typically have close relatives or even conspecifics among tropical Asian lineages. The

Australian Muellerargia timorensis is sister to M. jeffreyana, a Madagascar endemic, but probably reached the continent via Southeast Asia (where it still occurs today). Sicyos is represented in the Southwestern Pacific by S. australis , and two unnamed species, one of which is endemic to New Zealand, while the remaining c. 50 species of Sicyos occur in the Neotropics and on Hawaii.
The genus reached the Southwestern Pacific probably via long-distance dispersal by birds.
Origin and diversification of Holopogonoid
Lobeliaceae in Australia
Walsh, NG 1 , Knox, EB 2 , Albrecht, DE 3
1 Royal Botanic Gardens Melbourne, Australia;
3
2 Indiana
University, Bloomington, Indiana, USA; Alice Springs
Herbarium, Alice Springs, Northern Territory, Australia
Lobelia sect. Holopogon comprises 16 species, two of which have morphologically distinguished subspecies, thereby resulting in 19 terminal taxa that are all restricted to Australia. A chloroplast DNA-based phylogenetic estimate (using atp B, rbc L, and their intergenic region) is used to reconstruct the biogeographic history of this clade, which also includes the monotypic segregate genus
Colensoa from New Zealand. The floristically rich, southwestern region of Western Australia is the centre of diversity of this group, and also the inferred site of colonization from a climatically similar region in the
Western Cape of South Africa. One sub-clade of seven species is restricted to Western Australia except for the extension of L. heterophylla subsp. centralis into more or less adjacent areas of South Australia and the Northern
Territory and the establishment of L. andrewsii in New
South Wales and Queensland. A second sub-clade of eight species shows evidence of repeated migration and long-distance dispersal that allowed L. gibbosa to colonize all states, established L. browniana from South
Australia to Queensland, founded a common origin of L. dentata and L. simplicicaulis in Victoria or New South
Wales (with the latter dispersing to South Australia and
Tasmania), and extended L. rhombifolia to South
Australia, Victoria, and Tasmania. A third lineage established L. trigonocaulis in New South Wales and
Queensland and dispersed to New Zealand, where it diverged radically to form Colensoa physaloides .
Origin and diversification of Hemipogonoid
Lobeliaceae in Australia
Knox, EB 1 , Albrecht, DE 2 , Walsh, NG 3
1 Indiana University, Bloomington, USA;
3
2 Alice Springs
Herbarium, Alice Springs, Australia; Royal Botanic
Gardens Melbourne, South Yarra, Australia
The species richness and taxonomic diversity of the
Lobeliaceae in Australia make it a center of diversity, but biogeographic reconstruction using a chloroplast DNAbased phylogenetic estimate (from atp B, rbc L, and their intergenic region) indicate that the Lobeliaceae originated in another center of diversity, the Western
Cape of South Africa. The contemporary Australian diversity of lobeliads is derived from three independent colonization events plus three additional species that have become naturalized. The most ancient colonization was directly from the Western Cape to Western Australia
(with subsequent diversification to form 17 extant species; presented separately), as was the most recent colonization when the South African Lobelia anceps dispersed to Australia and thence to New Zealand. The three naturalized species ( Monopsis debilis ,
Grammatotheca bergiana , and Lobelia erinus ) are also
South African and were likely introduced as seeds in hay that accompanied cattle shipments, but L. erinus has commonly cultivated ornamental forms and may have escaped repeatedly. However, the largest group of hemipogonoid Lobeliaceae in Australia is more closely related to the New World clades (which were previously derived from South African ancestry). Most of the 30+
Australian species in this hemipogonoid clade are restricted to the eastern half of the country, but repeated north-south dispersal has created centers of diversification in Queensland and Victoria. Three old lineages reached the southwestern region of Western
Australia, but none of these lineages apparently diversified beyond a single species. This predominantly
Australian hemipogonoid clade is not restricted to
Australia, which is the source area for three dispersal events to New Zealand, two dispersal events to Asia, and one to South America.
Phylogeny and biogeographic patterns in Asian and
Australasian sedges in tribe Cariceae
Waterway, MJ 1 , Bruhl, J 3
Lange, P 5 , Ji, W 6
, Hoshino, T 2 , Wilson, K 4 , De
1 McGill University, Canada;
Science, Japan; 3
2 Okayama University of
University of New England, Armidale,
5
Australia; 4 Royal Botanic Gardens, Sydney, Australia;
Peter De Lange, Dept of Conservation, New Zealand;
6 Northwest Agricultural and Forestry University, China
Cyperaceae tribe Cariceae, a nearly cosmopolitan group with 2000+ species, is an ideal group for studying biogeographic patterns at the generic and species level.
These sedges are most common in temperate, boreal, and arctic habitats but also occur in subtropical areas where they are generally found at higher elevations.
Phylogenetic relationships among the species are increasingly better known, providing the opportunity to analyse their biogeographic relationships. Carex is by far the largest genus in the tribe and early molecular phylogenetic studies suggested that all of the other genera ( Cymophyllus, Kobresia, Schoenoxiphium , and
Uncinia ) were nested within it. Recent molecular phylogenies suggest an Eastern Asian origin for the group with the small Siderostictae clade (endemic to
Eastern Asia) sister to all other species, and Eastern
Asian species basal in two large well-supported clades that correspond to Carex subgenus Vignea and Carex subgenera Carex and Vigneastra . The phylogenetic positions and origins of the small monophyletic African genus Schoenoxiphium and the Caricoid clade (100+
229

species, comprising most unispicate Carex species plus
Cymophyllus , Kobresia and Uncinia ) are less clear. A large majority of species in tribe Cariceae occur in the
Northern Hemisphere, but there are also species radiations in the Southern Hemisphere. Our studies of more than a third of the species in the tribe suggest that most Australasian species are related to Eastern Asian species. In the largest clade (subgenera Carex and
Vigneastra ), native Australasian species follow one of three patterns: (1) endemics with Asian sister species nested within various larger groups with Asian or circumboreal distribution; (2) species found in both Asia and Australasia; and (3) endemics that are part of a large
Australasian (mostly New Zealand) radiation within the widespread Section Spirostachyae . The patterns are similar in subgenus Vignea with examples of the first two patterns in sections Glareosae , Holarrhenae , Ovales , and
Stellulatae plus two larger radiations, one within circumboreal section Heleoglochin and a second within a well-supported Australasian clade comprising species previously classified in four sections. In the Caricoid clade, a few alpine species are endemics of various origins, but Uncinia is a large Australasian (mostly New
Zealand) radiation. Biogeographic analyses suggest that the first two patterns likely resulted from recent long distance dispersal, with low levels of sequence divergence between Asian and Australasian species supporting this view. All four larger radiations include species from Australia and New Zealand with greater genetic differentiation among species within these groups than between continents in the first two groups. Dispersal in both directions between Australia and New Zealand is suggested by phylogenetic relationships in at least three of these major radiations. The overall pattern suggests early colonists in a few groups of tribe Cariceae radiated in Australasia forming most of the New Zealand and about half of the Australian Cariceae flora, but a few
New Zealand natives and about half of the Australian natives are more recent arrivals, many of which have diverged enough from their Asian relatives to be recognized as distinct species.
Phylogeographical and cytological studies of eastern
Asian
Carex
Hoshino, T 1 , Masaki, T 1 , Yano, O 2
1 Okayama University of Science, Japan; 2 University of
Tokyo, Japan
Carex L. is one of the largest genera of vascular plants and has more than 200 species in Japan. Many Japanese
Carex species seem to have adapted to various habitats in the Japanese archipelago. Carex oxyandra, C. conica , and C. blepharicarpa are perennial herbs that are widely distributed from Hokkaido to Kyushu in Japan. C. oxyandra is distributed from Sakhalin and Japan to Yu
Shan of Taiwan. C. conica is a common sedge in the forest understory and margins, widely distributed from
Hokkaido to Kyushu in Japan and in southern Korea. C. blepharicarpa is commonly found in alpine grasslands, forest understories, and along roadsides in mountaineous areas of Japan. The genetic variations of these plants that have adapted to the islands appear to be strongly affected by the geographical isolation events. These three species are considered to be ideal candidates for investigating the evolutional history of widespread herbaceous plants in
230
Japanese archipelago. Carex has diffuse centromeric chromosomes and a remarkable aneuploid series ranging from n = 6 to n = 66. Four aneuploids (2n=18, 20, 24, 26) are reported in C. oxyandra , nine aneuploids (2n = 32–
39, and 42) in C. conica , nine aneuploids (2n=26–33, and
41) in C. blepharicarpa . Intraspecific chromosome variations in Japanese Carex can be grouped into two types: (1) exemplified by C. oxyandra and C. conica , show no chromosome variation in chromosome number within populations, apparent correlations between geography and intraspecific aneuploidy, (2) illustrated C. blepharicarpa and C. duvaliana , have intraspecific aneuploid series within populations. We investigated the phylogeographic pattern and genetic diversity of the widespread Japanese sedges based on chloroplast DNA haplotypes and chromosomal variations. We found the geographical distribution of intraspecific aneuploids of
C. oxyandra and C. conica corresponded well with those of the haplotypes, and our results suggest that the genetic diversity and chromosomal variations in these species may have originated from contractions and expansions of geographical ranges affected by Quaternary climatic oscillations.
Leaving the dragons: cause and consequence of biogeographic range shift out of the southern Africa
Drakensberg Mountains in
Schoenoxiphium
Nees
(Cariceae, Cyperaceae)
Gehrke, B 1 , Martín-Bravo, S 2 , Muasya, M 3 , Luceño, M 2
2
1 Johannes Gutenberg-Universität Mainz, Germany;
Universidad Pablo de Olavide, Sevilla, Spain;
3 University of Cape Town, South Africa
The cosmopolitan tribe Cariceae (c. 2000 spp;
Cyperaceae) is poorly represented in sub-Saharan Africa and Madagascar, with only about 100 species. Having established itself in the area through several independent colonisation events at different times, diversification in most lineages is restricted. The monophyletic genus
Schoenoxiphium is, with more than 20 taxa, the largest and most widespread African clade in the Cariceae. Its present day distribution pattern could be the result of a complex biogeographic scenario and probably involves the interplay of processes such as hybridisation, shifts in morphology and possibly even polyploidisation (which is exceedingly rare in the Cariceae). Some of the more narrowly endemic species, for example, seem to show greater levels of cytological and morphological variation than those which have large distribution areas spanning thousands of kilometres across the continent, while others show the opposite patterns. Furthermore, several of the recently identified clades within Schoenoxiphium include species-complexes that are genetically and morphologically variable, indicating gene-flow and incomplete divergence. We present results of molecular phylogenetic and dating analyses, ancestral area reconstruction, and cytological and morphological investigations. By combining these different lines of evidence we address the causes and consequences of geographic range shifts in Schoenoxiphium . Preliminary analyses indicate that Schoenoxiphium originated in the alti-montane grasslands of the Southern African
Drakensberg Mountains. These grassy slopes represent the current centre of species diversity and are also where most diversification occurred. The radiations of species

appears to have coincided with periods of geological change, leading to the uplift of the area, and of climatic change, both of which are likely to have (simultaneously) influenced past plant distributions.
Phylogeny, phylogeography, and chromosomal evolution in
Carex
section
Spirostachyae
and allied sections
Luceño, M 1 , Escudero, M 1 , Jiménez-Mejías, P 1
1 Botany Area, Dept of Molecular Biology and
Biochemical Engineering, Pablo De Olavide University,
Spain
Carex (Cyperaceae) with ca. 2000 species has undergone the most remarkable radiation in the temperate regions of
Northern Hemisphere. In the last years, caricologists have attempted to disentail which factors or mechanisms could explain such extraordinarily high species diversification. In order to achieve this goal, we display a study case in section Spirostachyae and allied sections
Ceratocystis and Rhynchocystis . Ecological, biogeographical and cytogenetic factors seem to play an important role in the differentiation and speciation process. In this way, the three phylogeny sections display a mainly biphyletic arrangement which matches ecological, morphological and geographical signal. The molecular differenciation found in sect. Spirostachyae is higher than those observed in Ceratocystis and
Rhynchocystis . The two main Spirostachyae clades fit to acidofilous species with frequently higher size and basofilous and smaller species size. The two main clades of Ceratocystis show soil type signal and morphological differences in the utricles (perigynia). Finally,
Rhynchocystis clades have biogeographical (tropical and subtropical Africa vs. the Mediterranean and Eurasian regions) and morphological differences (species size).
All the three sections exhibit disjunct and widespread distribution what entail relatively frequent transatlantic and transhemispherical dispersals. In addition, studies at species level ( C. helodes, C. extensa and C. lepidocarpa s.l.) suggest that isolation by distance has a crucial role in the population differentiation process. Carex species display holocentric chromosomes which are characterized by diffuse centromere and it is supposed to be an important driver of chromosome number evolution by fission, fusion and translocation events. Although section Spirostachyae displays neither shifts in its karyotypic equilibrium nor evidences of chromosome selection, the relation between the chromosome number variation and relative time of coalescence from molecular makers has been proved, which suggest that chromosome rearrangements may promote the differentiation and speciation process. Soil type, high ability of long distance dispersal and frequent chromosome rearrangements seem to be some of the most important factors to explain the high species richness in the genus
Carex .
Phylogeography of bipolar and arctic-alpine Cariceae
(Cyperaceae): origins, refugia and systematics
Starr, JR 1,2
C
W
5 , Escudero, M
1,2
, Bruederle, LP
7 , Hipp, A.L
, Villaverde, T 1,2
3
6
, Luceño, M 4 , Brochmann,
, Martín-Bravo, S
, Bakke Westergaard, K 5,7
4 , Sawtell,
1
4
University of Ottawa, Canada;
Nature, Canada; 3
2 Canadian Museum of
University of Colorado Denver, USA;
Pablo De Olavide University, Seville, Spain; 5 University
7 of Oslo, Norway; 6 The Morton Arboretum, USA;
University of Tromsø, Norway
Although tribe Cariceae is remarkable for its exceptional diversity (ca. 2100 species) and peculiar cytology
(agmatoploidy and symploidy), it is also notable for its biogeography. Almost every classic biogeographic pattern recognised in plants from Gondwanan through to amphiatlantic and circumpolar distributions are seen in at least one species or species group, a fact that lead Léon
Croizat to remark that phytogeography could be taught using only examples from this tribe. In this study, we examine the systematics and phylogeography of some of the most widespread species on earth: the six bipolar
Carex species ( C. maritima, C. capitata, C. microglochin, C. macloviana, C. magellanica, C. canescens ) that range from the Arctic south to Tierra del
Fuego and three arctic-alpine Cariceae ( C. nardina, C. rupestris, Kobresia myosuroides ) that share the range of the bipolar carices in the Northern Hemisphere. To determine whether their shared range is an artifact of an unresolved taxonomy, evolutionary convergence or the influence of common historical events such as glaciation and long-distance dispersal we examined the phylogeography of these species across their entire ranges using Bayesian analysis and statistical parsimony of multiple chloroplast and nuclear (ITS, microsatellites) regions. Our analyses suggest that at least two taxa, the bipolar C. microglochin and C. capitata represent species complexes of either closely related taxa ( C. capitata ) or distantly related species ( C. microglochin ) suggesting extraordinary morphological convergence within the bipolar bog-habitat group Carex sect. Leucoglochin .
Nonetheless, both groups still contain bipolar taxa whose genetic diversity is low and whose haplotype networks, like the other bipolar Carices, suggest recent dispersal from the Northern Hemisphere to South America either by mountain hopping ( C. microglochin ) or by an immediate and direct dispersal by bipolar migratory birds. Analyses also suggest that despite sharing a common distribution, current species ranges in the
Northern Hemisphere reflect dispersal from multiple, often different refugia in Eurasia and North America
(southern Rockies, Beringia) and the presence of relict taxa whose current distribution predates the last glacial maximum in the southern Rocky Mountains.
Chromosome evolution and its effects on
1 diversification in sedges (
Carex
: Cyperaceae)
Hipp, A 1 , Roalson, E 2 , Chung, K-S 1 , Escudero, M 1
The Morton Arboretum, Lisle, USA; 2 Washington State
University, Pullman, USA
Chromosome rearrangements play a crucial role in speciation and species evolution. They drive lineage and genetic diversification through the production of hybrid incompatibilities and suppression of recombination. Yet chromosome rearrangements are neither sufficient nor necessary conditions for speciation. Within the sedge family Cyperaceae (ca. 5000 species worldwide), the genus Carex (ca. 2100 species worldwide) has undergone a dramatic chromosomal radiation. Chromosome number
231

changes in the genus are largely due to fissions, fusions, and translocations, not duplications or deletions.
Consequently, there is a potential for rapid rearrangement of linkage groups without incurring immediate reproductive barriers. Recent work demonstrates that chromosome divergence within species drives genetic diversification of populations, while still permitting species coherence across wide geographic ranges.
Chromosome rearrangements act in concert to reduce gene flow among populations. Thus, it is not the presence of a single rearrangement that drives population divergence. Rather, genetic diversity within species is an increasing function of the number of chromosome rearrangements differentiating populations. Among species, chromosome evolution proceeds rapidly, in some cases more rapidly than lineage diversification, such that phylogenetic signal is absent in for example the species of Carex section Ovales . Rates of chromosome evolution shift among clades within the genus, and cladogenesis is often associated with a shift in equilibrium chromosome number ranges. This combination of high evolutionary rates and shifting dynamics of chromosome evolution reduces the certainty of ancestral chromosome number reconstructions at most internal nodes of the genus.
However, this high rate of evolution also provides a unique opportunity to study the possibility for natural selection to act on chromosome number, and in fact there is evidence of environmental selection acting on chromosome number, perhaps through its effect on recombination rates. Genome size, on the other hand, apparently evolves independently of chromosome number and at an approximately constant rate across the genus. The relationship between chromosome number and genome size appears to shift from negative at deeper phylogenetic depths to positive at shallower depths, suggesting shifting dynamics of chromosome evolution in different clades or a lag time in the normalization of genome size with chromosome number changes. One outstanding question remains: how does the rate of chromosome evolution influence lineage diversification?
In this talk, we will present evidence from a densely sampled clade ( Carex subgenus Vignea ) based on 4 low copy nuclear genes and nuclear ribosomal DNA to address the question of whether increased rates of chromosome evolution lead to increased speciation rates.
The fossil record of the earliest land plants: evidence for the algal/bryophyte transition?
Wellman, CH 1 , Graham, LE 2 , Lewis, LA 3 , Taylor, WA 4
1 University of Sheffield, UK;
Madison, USA; 3
2 University of Wisconsin,
University of Connecticut, USA;
4 University of Wisconsin, Eau Claire, USA
It is generally assumed that terrestrial environments harboured microbial communities long before the appearance of land plants sometime in the Mid
Palaeozoic. Until recently, however, evidence (fossil, sedimentological and geochemical) for these biotas was rare. Recent palynological investigation of ancient
232 terrestrial deposits has yielded abundant and exciting new evidence for early terrestrial life. This includes evidence from the Nonesuch Shale of the USA (ca. 1100
Ma), the Torridonian of Scotland (ca. 950 Ma), and the pre-Mid Ordovician of Oman. The palynomorph assemblages yield diverse assemblages of unicellular prokaryotes and eukaryotes (sheathes, filaments, tubular structures and sphaeromorphs – including encysted forms and clusters). Many forms are quite large, including sphaeromorphs exceeding 400 µm in diameter, and others exhibit complex wall structure. Tantalizing structural elements indicate the presence of larger, possibly multicellular organisms. However, the biological affinities of these palynomorphs are largely unknown. They represent allochthonous elements that accumulated in aquatic environments, and probably in the main represent free-living benthic and planktonic freshwater communities. However, some elements may have been transported into these depositional environments from terrestrial subaerial habitats. The
Torridonian also preserves abundant sedimentary structures interpreted as representing subaerial microbial mats, and the palynomorph assemblages from these sediments contain autochthonous wefts of these mats
(associated with various cyanobacteria). Thus these palynomorph assemblages are revealing the nature of the complex microbial communities of organisms that inhabited terrestrial (subaerial and freshwater aquatic) environments that existed prior to the appearance of land plants. By the Mid Ordovician dispersed land plant spores (often termed cryptospores) appear in palynological preparations. They occur in large numbers worldwide, forming similar assemblages that persist for some 30 million years. These dispersed spores are considered to represent the first land plants
(embryophytes). They are interpreted as plants at a
‘bryophyte-like’ grade or organization, that were probably ecological generalists and hence cosmopolitan.
By the Silurian there is megafossil evidence for polysporangiate plants. There was a major turnover in floras as the polysporangiates (vascular plants and their immediate precursors) outcompeted the ‘bryophyte-like’ plants. The latter were displaced into the ecological
‘nooks-and-crannies’ while the former underwent an adaptive radiation. This talk will critically review the fossil evidence for the earliest land plants and their algal precursors and describe a collaborative project aimed at resolving some current controversies. These include: (i) the nature and biological affinities of the earliest terrestrial microbial fossils as revealed by taphonomic experiments on extant material; (ii) the affinities of
Cambrian palynomorphs purported to represent an early appearance of land plant spores; (iii) the nature of the earliest embryophytes and their relationship to extant liverworts.
The timing of evolution across land plants – recent results
1
Bell, C 1
University of New Orleans, USA
Recent molecular phylogenies have provided support for the hypothesized early radiation of embryophytes (land plants). In combination with the available fossils for land plants recently proposed ‘relaxed clock’ methods,

these molecular data can be used to estimate the ages of differentiation and diversification of plant lineages. The inherent disparity in taxon sampling for such large-scale phylogentic problems makes the placement of fossil constraint with any precision problematic, potentially leading to great uncertainty in our estimates. In this talk I will highlight some advantages/disadvantages of recently proposed methods that deal with this uncertainty in fossil placement. Likewise, I will also explore some potential problems associated with using estimates from such analyses as external calibrations in other studies that might not have readily available fossil data.
Divergence timing and molecular rates across land plants: the influence of differential substitution rates
1 and fossil constraints
Magallon, S 1 , Hilu, K 2 , Quandt, D 3
Universidad Nacional Autonoma de Mexico;
Tech, USA; 3
2 Virginia
Friedrich-Wilhelms Universität, Germany
The evolution of land plants is among the major events in the history of life on Earth. In addition to causing major global biogeochemical changes, it had determinant consequences in the diversification of terrestrial biota and the development of terrestrial ecosystems, eventually leading to present-day biomes. The temporal framework of land plant evolution is crucial for an understanding of rates of phylogenetic diversification, morphological evolution, and biogeographical and ecological history.
Paleobotanical data have consistently indicated the onset of land plant evolution during the Middle and Late
Ordovician. Recent discoveries document the presence of plants on land in the earlier part of the Middle
Ordovician, between 468 and 472 Mya, earlier than suggested by previous paleobotanical evidence. We conducted relaxed molecular clock analyses to investigate the timing of phylogenetic divergence and rates of molecular evolution across land plants, evaluating the effect of molecular markers with different substitution rates, and of fossil-derived age constraints.
We executed Bayesian uncorrelated lognormal relaxed clocks on a representation of all major land plant lineages
(liverworts, mosses, hornworts and tracheophytes) plus an algal outgroup, using nucleotide sequences of four highly-conserved ( atp B, psa A, psb B and rbc L) and one rate-variable ( mat K) protein-coding chloroplast genes, 27 fossil-derived age constraints, and a calibration for the land plant crown node distributed between 470 and 475
Mya. In addition to estimating absolute times and molecular rates across land plants using the five genes in combination, we explored the times and rates derived from the highly-conserved genes, and matK, separately.
Whereas the ages estimated by the two types of genes are very similar and highly correlated, the respective molecular rates differ, as expected, but are nevertheless strongly correlated. To address the question if observed similar ages and correlated rates were a consequence of the implemented fossil constraints, we investigated if excluding these constraints would result in a substantial decrease in the similarity and correlation between the ages and rates derived from the two types of genes.
When fossil constraints were excluded, estimated ages differed substantially (being usually younger) from those in fossil-constrained analyses, irrespective of whether the five combined genes, the four conserved genes, or mat K, were used. However, fossil-constrained and unconstrained absolute rates were very similar. In spite of being substantially different from fossil-constrained age estimates, unconstrained age estimates derived from the conserved genes and from matK were very similar.
Unconstrained estimations slightly increased the dissimilarity between conserved genes and mat K absolute rates, but nevertheless, the correlation between the two remained high. These results document correlated molecular rates across land plant lineages, as well as the effect of fossil-derived constraints in relaxed clock estimation of times and rates. In conclusion, molecular markers with substantially different substitution rates can provide very similar ages and strongly correlated molecular rates across land plants, regardless of whether fossil constraints are implemented.
However, whereas the inclusion/exclusion of fossil constraints has important consequences on the absolute ages of many nodes, it has little impact on the absolute rates of most branches.
Exposing global patterns of diversification within leptosporangiate ferns
Schneider, H 1 , Pryer, K 2
1 University of North Carolina Wilmington, USA; 2 Duke
University, Durham, USA
The fossil record indicates that leptosporangiate ferns originated in the Carboniferous and subsequently underwent three successive radiations, the last of which began in the Cretaceous and resulted in the bulk of extant fern diversity. Recent analyses, integrating fossil and living data, confirmed the existence of this third radiation, demonstrating that most of the approximately
9000 living fern species arose in the last 150 million years. There is no indication, however, that all of this diversity appeared simultaneously. Instead, focused studies have provided a variety of lineage-specific age distributions. But are there underlying patterns? Was diversification in separate lineages correlated? Through a broad-scale analysis, we aim to address these questions and to evaluate the effects of major biotic and abiotic events in Earth’s history on the diversification of leptosporangiate ferns.
Understanding biases in molecular dating analyses: lessons from campanulid angiosperms
Beaulieu, JM 1 , Donoghue, MJ 1
1 Dept of Ecology and Evolutionary Biology, Yale
University, Connecticut, USA
For flowering plants, the ages of clades estimated from molecular phylogenetic analyses have not always corresponded well with the accepted fossil record. In particular, the application of molecular clock methods has tended to yield older dates, in many cases much older than has seemed credible based on the stratigraphic record. As methods have further relaxed the assumptions of substitution rate inheritance (e.g. uncorrelated relaxedclocks) and acknowledge the uncertainty in a fossil age by treating fossil calibrations as probabilistic priors, the gap between the stratigraphic record and molecular age estimates seems to only get wider. It may be that these
233

older molecular dates are indeed real and are providing a new understanding of key events in flowering plant evolution. However, it is still unclear how uncorrelated relaxed-clock models perform in the presence of molecular rate shifts between lineages and how the placement of fossil calibrations influences their performance in these situations. Here we study these issues in detail by focusing on the Campanulidae, a large angiosperm clade that has been the subject of a recent molecular phylogenetic analysis. With respect to the dating, the campanulids, and the major lineages within the clade, are inferred to have existed far back into the
Cretaceous, possibly too far based on biogeographic and morphological analyses. We explore the possibility that these discrepancies reflect potential biases in the divergence time estimates of campanulid lineages. We use a simulation approach to assess the impact of lineage-specific rate heterogeneity on molecular age estimates. We also assess how biases in the fossil record can enhance the impacts of lineage-specific rate heterogeneity in divergence time analyses. These simulations suggest that in certain situations current methods are unable to cope with extreme differences in molecular rate and older age estimates are often the consequence. Our study also demonstrates how inferences about biogeographic history and morphological evolution might be useful in pinpointing instances where molecular age estimates are potentially misleading.
‘Out-of-Africa' dispersal of tropical floras during the
Miocene climatic optimum: evidence from
Uvaria
(Annonaceae)
Zhou, L 1
RMK 2
, Su, Yvonne CF 2 , Thomas, DC 2 , Saunders,
1 State Key Laboratory of Emerging Infectious Diseases and Dept of Microbiology, Li Ka Shing Faculty of
2 Medicine, The University of Hong Kong; School of
Biological Sciences, The University of Hong Kong,
China
African–Asian disjunctions are common in palaeotropical taxa and have been the focus of considerable biogeographical research. Alternative hypotheses for these biogeographical disjunctions have been proposed, including ‘rafting’ on the Indian tectonic plate, dispersal via Eocene ‘boreotropical’ forests, and long-distance dispersal. These hypotheses are tested using the angiosperm genus Uvaria (Annonaceae), with divergence times estimated using a Bayesian relaxed molecular clock. Uvaria originated in continental Africa, ca. 31.6 Ma, and migrated into Asia in the early to middle Miocene. ‘Rafting’on India and the Eocene boreotropical hypotheses are rejected due to incongruence with divergence times, and the genus is shown to be dispersed by primates, making it an unlikely candidate for transoceanic dispersal. Here we show that dispersal is likely to have occurred via boreotropical forests associated with the late Middle Miocene thermal maximum. This is the most convincing evidence yet for such a dispersal route for a tropical plant group.
Hornworts: fitting the puzzle together
, Villarreal, JC 2
1
Renzaglia, K 1
Dept of Plant Biology, Southern Illinois University
Carbondale, USA; 2 University of Connecticut, USA
The long and spirited debate over the placement of hornworts in the green tree of life is now settled. This small plant phylum is the sister group to tracheophytes.
But this newly-resolved position among embryophytes only raises more questions about the transition from gametophyte to sporophyte dominance. Indeed, there are no manifest structural features of this plant phylum that support this key position among plants. The signature morphological traits of the group are unparalleled among extant plants. The chloroplast of many hornworts is large, solitary and contains a well-defined pyrenoid seen in no other land plant. Hornworts are the only land plants that have no evidence of conducting tissue in either the sporophyte or gametophyte generation. The sporophyte is a single sporangium that continues to grow from the base, not the apex, throughout its life cycle. Spores are incrementally produced and released from the tip toward the base. The extensive columella in the sporophyte suggests affinity with early tracheophytes such as
Horneophyton but irrefutable hornwort spores only date back to the Cretaceous. Stomata are found in the sporophyte of approximately half of the known species but they are unable to respond to environmental or hormonal signals; once open they remain so. The placenta has gametophytic wall ingrowths only, a feature rare among embryophytes. The gametophyte resembles that of simple thalloid liverworts but there are no organized external appendages. The ubiquitous internal cyanobacterial symbiont is unknown among plants and gametangia that are sequestered inside the thallus hint of fossil and living pteridophytes. Over the past decade, relationships among hornworts have also been resolved.
We now recognize approximately 200 species in 14 genera, with Leiosporoceros the sister taxon to the remaining hornworts. Morphological boundaries between genera are often blurred due to the mosaic of shared characters. The molecular revolution has led to unpredicted and inexplicable relationships among hornwort taxa and with other plants.This presentation will explore the new phylogenetic hypotheses related to hornworts, and identify morphological support for these interrelationships. Morphological change within hornworts also will be examined. Areas of future study will be discussed that likely will yield additional structural clues to the evolution of polysporangiate from monosporangiate embryophytes. Does the fossil record further illuminate these critical structural transformations? New insights emerge with a fresh, unbiased re-examination of this homogenous and unique plant clade.
Evolution of sex in hornworts: from genes to genomes
Villarreal, JCA 1
Bainard, J 4
, Forrest, LL 1
, Goffinet, B 1
, Cargill, DC 2 , Wickett, N 3 ,
234

1 University of Connecticut, Storrs, USA; 2 Centre for
Australian National Biodiversity Research, Canberra,
Australia; 3 Pennsylvania State University, USA;
4 University of Guelph, Ontario, Canada
Among bryophytes, hornworts compose the sister group to vascular plants and may thus be critical for understanding the evolutionary transition of land plant life cycles from haploid-dominant in bryophytes, to diploid-dominant in vascular plants. Hornworts comprise only ~200 species, and are thus much less speciose than liverworts (7000 sp.) or mosses (11000 sp.). They are characterized by low chromosome numbers of 4 + 1 sex chromosome in dioecious taxa and 5 to 7 chromosomes in monoecious taxa. Neopolyploidy is rare in hornworts
(c. 3%) but rather rampant in other bryophytes. Evolution of sexual condition in bryophytes is thought to evolve from dioecy to monoecy following chromosome doubling. Given the low incidence of polyploidy in hornworts we address the evolution of sexual condition in hornworts using phylogenetics tools, genome size estimates and preliminary transcriptomic data from
Nothoceros aenigmaticus . To generate the most complete hornwort phylogeny to date, we have sequenced 30% of total hornwort diversity for two loci. We used phylogenetic reconstructions to analyze evolution of sexual condition and mapping genome size estimates from representatives of each major clade.
Leiosporoceros, the earliest diverging hornwort is dioecious. Monoecy is the predominant condition (60 %) in our data set. Transitions from dioecy to monoecy within clades are rare; although exceptions occur, such as within the family Dendrocerotaceae. In addition, we have generated a complete species-level phylogeny for
Nothoceros , which emphasizes a transition from monoecious taxa in temperate South America and the
Neotropics, through dioecious taxa in high elevation tropical America, to asexuality in the Southern
Appalachian Nothoceros aenigmaticus . This is the first phylogenetic report of a transition from monoecy to dioecy in bryophytes, in contrast to the widely accepted evolution of monoecy from dioecy following genome doubling. Chromosome data and genome size estimates in hornworts are sparse. A putative correlation between sperm cell size and genome content suggest that most hornworts have similar nuclear content. To assess the apparent lack of neopolyploids in hornworts we have obtained genome size estimates using flow cytometry for most species within Nothoceros and across major clades of hornworts. Our preliminary results suggest that hornworts have indeed similar genome sizes independent of sex condition. In fact, the genome size estimates for hornworts are much smaller than other bryophytes. In addition, we have initiated transcriptomic work on gametophyte generation of several hornwort genera.
High-throughput second-generation shotgun genomic and transcriptomic sequencing will provide more insight into whether hornworts are in fact paleopolyploids.
Neopolyploid species have undergone a recent increase in genome size, and have higher chromosome numbers.
Paleopolyploids on the other hand are more difficult to detect, because they undergo haplodization, wherein many genes return to low or single copy numbers, and chromosome number is likewise reduced. Our genetic and transcriptomic analyses will help elucidate the evolution of hornwort genomes, particularly regarding levels of paleoploidization. The lack of apparent polyploidy in most hornwort species poses intriguing questions on the evolution of sexual condition in hornworts and provides a platform to address the evolution of mating system in bryophytes.
1
Hornworts: a new look at stomatal evolution
Pressel, S 1 , Renzaglia, K 2 , Duckett, J
Natural History Museum,UK; 2
1
Plant Biology Dept,
Southern Illinois University, Carbondale, USA
The presence of stomata in the sporophytes of hornworts has been a pivotal morphological character used alongside sequencing in drawing up current total evidence phylogenies of land plants that now place hornworts sister to tracheophytes. Embedded in these phylogenies is an assumption of stomatal homology across all groups of embryophytes. However, our studies on responses of stomata of different ages along mature hornwort sporophytes of the genera Anthoceros ,
Paraphymatoceros and Phaeoceros to desiccation, darkness and abscisic acid (ABA), together with comparative observations on the development of intercellular spaces in hornwort sporophytes and tracheophyte leaves, point to a very different scenario.
Using cryo-scanning electron microscopy (SEM) and xray microanalysis we found, in striking contrast to tracheophytes, no correlation between the guard cell potassium content and aperture in hornworts for any of the treatments, nor in stomata of different ages. Stomata open early in sporophyte ontogeny and thereafter remain so as a consequence of changes in the guard cell walls rendering these inelastic. Differently from those in tracheophytes, which are gas-filled from the outset, in hornworts the intercellular spaces are initially filled with mucilage; this is gradually replaced by air but only after the stomata have opened and at the level of the dehiscence grooves. Intercellular spaces are absent in
Dendroceros, Nothoceros , Megaceros and Notothylas, four derived hornwort genera that also lack stomata.
Unlike the sporophytic stomata, the ‘guard cells’ surrounding the gametophtic slime pores in hornwort thalli undergo repeated divisions following the ingress of cyanobacteria. In the light of this major ontogenetic difference it would seem highly unlikely that the gametophytic slime pores in hornworts ever functioned as stomata. The absence of evidence for a potassiumregulating mechanism in the stomatal guard cells of hornworts, their failure to respond to the environmental cues normally associated with closure in tracheophytes and the replacement of intercellular mucilage with air post their early opening indicate that their principal role is facilitation of sporophyte desiccation leading to dehiscence and spore dispersal. These features of the stomata and the very different origins of gas-filled intercellular spaces cast serious doubts on stomatal homology between hornworts and tracheophytes.
Alternatively, when taken together with the demonstration of a similar function for the pseudostomata in the walls of Sphagnum capsules and the presence of stomata on the sporangia of the earliest fossil plants in the mid-Paleozoic, these novel hornwort data add weigh to an alternative notion that stomata first appeared in land plants as structures that facilitated sporophyte dehiscence when CO
2
levels where high and planate leaves had yet to evolved and only subsequently,
235

as CO
2
levels decreased and temperatures rose in the late-Paleozoic, acquired their role in the regulation of gaseous exchange and cooling of planate leaves.
Fungal symbioses in hornworts
Duckett, J 1 , Pressel, S 1 , Ligrone, R 2 , Bidartondo, M 3
1 Natural History Museum, UK; 2 Dipartimento di Scienze ambientali, Seconda Università di Napoli, Italy;
3 Imperial College London, UK
Whereas the ubiquitous occurrence of cyanobacterial endosymbionts in hornworts was well documented in the nineteenth century it was not until 1988 that Ligrone first demonstrated an unequivocally symbiotic relationship with a fungus in an ultrastructural study of Phaeoceros laevis . This was subsequently confirmed by the experimental establishment of an arbuscular mycorrhizalike symbiosis between Anthoceros puncatatus and
Glomus claroideum by Schußler in 2000. Our current cytological and molecular studies are now providing some surprising discoveries about the extent and nature of the fungal associations in hornworts. Looking across the 14 hornwort genera these associations are absent in the basal taxon Leiosporoceros dussii and from the derived clade containing Nothoceros and Megaceros , two genera that grow in extremely wet habitats, and the epiphytic and epiphyllic genus Dendroceros . Associated fungi have also not been found in Notothylas but we have yet to examine fresh materials of this speciose genus as is the case for Folioceros , Hattorioceros , Mesoceros , and
Sphaerosporoceros . In contrast they are present in all the species examined to date in Anthoceros,
Paraphymatoceros, Phaeoceros, Phaeomegaceros and
Phymatoceros . Cytologically the associations in these five hornwort genera share features on the one hand with the Mucormycotina symbionts described in Treubia and
Haplomitrium viz. extracellular hyphae and thick-walled spores in copious host-produced mucilage and on the other with the widespread Glomerales in thalloid liverworts viz. intracellular arbuscules and vesicles.
Molecular analyses are beginning to reveal surprising diversity in the fungi. This includes Glomerales and
Diversisporales in Anthoceros laminiferus and
Phaeoceros carolinianus (both from New Zealand),
Archaeosporales in A. husnotti and P. laevis (both from
UK) and Mucormycotina in Paraphymatoceros and
Phaeoceros carolinianus (Australia and New Zealand).
The discovery of three very different fungi in Phaeoceros suggests much less host specificity than in thalloid liverworts and opens the way to novel experiments on possible functional differences between the mycobionts.
Physiological ecology of the carbon concentrating mechanism in hornworts
Griffiths, H 1
1 University of Cambridge, UK
The primary carboxylase Rubisco has a low selectivity for CO
2
, and slow turnover rate, under modern atmospheric conditions, as well as a propensity to engage with O
2
as a competitive substrate. A range of carbon concentrating mechanisms (CCM) have evolved in both aquatic and terrestrial habitats to provide a physiological
236 fix for these problems, by locally concentrating CO
2 around Rubisco. The biochemical metabolism associated with C
4
and CAM pathways, and their respective spatial and temporal solutions to the decarboxylation of organic acids, are well defined. In contrast, biophysical CCM systems are found in cyanobacteria, many eukaryotic aquatic organisms and hornworts, based on active transport of bicarbonate and CO
2
. Under low CO
2 conditions for growth, Rubisco is packaged into subcellular compartments, namely the prokaryotic carboxysome, or eukaryotic pyrenoid. The molecular basis to this mechanism is now relatively well defined for cyanobacteria, as are the structural determinants and supramolecular complexes between Rubisco and carbonic anhydrase, associated with the carboxysome.
Alternatively, the eukaryotic CCM in algae and hornworts is normally associated with a chloroplast pyrenoid, for which the molecular physiology is less clearly defined, although associations between Rubisco small subunit and pyrenoid formation are promising
(Genkov et al. 2010). The paper will review the current understanding of biophysical CCM systems, their likely origins and diversity. For the hornworts, we have a limited dataset currently on the physiological ecology of the CCM. Preliminary observations suggest that for a non-ventilated, planar thallus, diffusive limitations arise because of surface water and dense internal tissues, to allow a CCM to provide a competitive advantage in terms of carbon gain for hornworts, as compared to some liverworts (Meyer et al. 2008). Phylogenetic comparisons will be made between potential CCM components and pyrenoid distribution in hornworts, to consider whether the CCM is a basal or derived condition. The implications of this study will help to resolve the question whether the CCM in hornworts represents a relict of an algal ancestor, and is possibly a palaeohistorical remnant of early land plant physiology.
Alternatively, if the CCM and pyrenoid are relatively modern evolutionary developments, there may be lessons for the introduction of a CCM into higher plants.
1
Hornwort diversity and conservation in Asia with special reference to Indian subcontinent
Singh, DK 1 , Asthana, AK 2
Botanical Survey of India, Kolkata, India; 2 National
Botanical Research Institute, Lucknow, India
The hornworts are a key group representing an important milestone in the initial journey of evolution of land plants. Yet, the paucity of national and regional floras and taxonomic revisions has considerably limited our knowledge on their diversity and distribution. Currently the group is estimated to comprise 200–250 species belonging to 14 genera the world over, distributed chiefly in tropical Asia, the Indian subcontinent, tropical
America, tropical Africa and the Pacific islands The
Indian subcontinent harbours about 49 species, in seven genera. Out of this 41 species belonging to six genera, viz. Anthoceros L., Folioceros D.C.Bharadwaj,
Hattorioceros J.Haseg. Megaceros Campb., Notothylas
Sull. and Phaeoceros Prosk., occur in India, which is the highest for any country in the world. Dendroceros Nees, represented by D. barbonicus Steph. in Sri Lanka, is the only genus in the region not recorded from India so far.
The genera Folioceros and Notothylas are considered to

have originated and diversified in India. However, the above figures could be a gross underestimate as these are based on limited bryological explorations and taxonomic studies on the group in countries like Bangladesh,
Myanmar and Sri Lanka. This gap in our current taxonomic knowledge base of the group, coupled with lack of trained local bryologists, has considerable implication on their assessment and conservation. In the present state of our knowledge, a number of species from the subcontinent like Folioceros mangaloreus (Steph.)
D.C.Bharadwaj, F. satpurensis (K.P. Srivast.)
D.C.Bharadwaj & K.P. Srivast., Notothylas nepalensis
Udar & D.K.Singh etc., are known through their type collection only, while Anthoceros alpinus Steph.,
Hattorioceros striatisporus (J.Haseg.) J.Haseg.,
Notothylas khasiana Udar & D.K.Singh, Phaeoceros parvulus (Schiffn.) J.Haseg., etc. are known from just two locations widely separated from each other. More explorations are therefore needed to not only unveil the total diversity of the group in the subcontinent, but also to document rare and threatened species and evolve conservation strategies. In situ conservation of rare and threatened species in ‘Species Specific Sites (SSS)’ and creating general awareness about high ecological and phylogenetic significance of the group is suggested with couple of examples.
Choosing and using a plant barcode
Hollingsworth, P 1
1 Royal Botanic Garden Edinburgh, UK
DNA barcoding aims to establish a central community resource for large scale use of DNA sequences for organismal identification and taxonomic clarification.
The approach was pioneered in animals using a portion of the cytochrome oxidase (CO1) mitochondrial gene and has led to a large number of studies applying the technique to distinguish among animal species. In plants, establishing a standardized DNA barcoding system has been more challenging. In this talk I will (1) review the process of selecting and refining a plant barcode, (2) evaluate the discriminatory power of the approach and the reasons underlying discrimination success and failure, (3) describe some applications of the approach and summarise major emerging projects, and (4) outline some research requirements and tool development required to take DNA barcoding forward in plants.
Environmental barcoding as a key to past and present ecosystem dynamics in the Arctic
Brochmann, C 1
Boessenkool, S 1
A2 2
, Bellemain, E 1
, Epp, LS 1
, Gussarova, G 1 ,
, Ecochange/BarFrost Teams,
1 National Centre for Biosystematics, Natural History
Museum, University of Oslo, Oslo, Norway; 2 Laboratoire d’Ecologie Alpine, CNRS, Université Joseph Fourier,
Grenoble, France
Recent advances in DNA sequencing technology coupled with bioinformatics-based development of mini-barcodes for species identification are opening up a novel research avenue for analyzing degraded DNA in environmental samples. We demonstrate that this approach is powerful both for assessing modern biodiversity from soil samples as well as for reconstruction of past ecosystems from
DNA preserved in arctic permafrost sediments (Mol.
Ecol. Res. 10: 1009-1018, 2010). Such reconstructions can be used to address past species turnover dynamics, niche stability in time and space, and backward testing of predictive species distribution models. For vascular plants, which are analyzed in the EU-funded
ECOCHANGE project, we use the P6 loop in the chloroplast trn L intron, a short (13–158 bp) and variable region with highly conserved flanking sequences. A trn L reference library was constructed by sequencing more than 1600 recently collected samples of 842 species, representing all widespread and 8260; or ecologically important taxa of the species-poor Arctic flora. The P6 loop allowed identification of most genera and one-third of the species, thus providing much higher resolution than pollen records. We then tested whether degraded
DNA in modern soil mirrors the above-ground vegetation in arctic plant communities. We found that the soil DNA diversity is highly consistent with plant functional and structural diversity estimated from conventional vegetation surveys. In the Research Council of Norwayfunded BarFrost project, we design new mini-barcoding markers for other groups of organisms (bryophytes, fungi, insects, springtails, vertebrates) using new bioinformatics tools, and test whether these markers are suitable for amplification of degraded DNA in environmental samples. New DNA reference libraries are constructed for Arctic as well as north boreal taxa. In addition to modern soil, both projects assess past diversity by amplifying DNA from more than 600 permafrost soil samples collected throughout the Arctic, ranging in age from 10 000 to several hundred thousand years. The results so far indicate that although the level of detection varies among organism groups, our approach enables us to reconstruct diverse communities through time.
DNA barcodes for Stipoid grasses: developing a better tool for biosecurity
Udovicic, F 1 , Syme, AE 1 , Murphy, DJ 1 , Stajsic, V 1
1 Royal Botanic Gardens Melbourne, Australia
Some of the most invasive weeds in Australia are exotic
Stipoid grasses, degrading pastures, injuring livestock and invading native grasslands. Two species have been recognised as Weeds of National Significance: Serrated
Tussock ( Nassella trichotoma ) and Chilean Needlegrass
( Nassella neesiana ). Together with other exotic Stipoids, these cost Australia tens of millions of dollars each year and could potentially cover hundreds of millions of hectares. Uncontrolled, it has been estimated that they could cost Australia half a billion dollars over the next 30 years. It is therefore of utmost importance to be able to identify with confidence any imported plant material that may be a weed species. Accurate identification, however, often relies on the floral characteristics, which may not be available. Using molecular characters such as DNA barcodes is a new way to identify grass species and to
237

help prevent the importation and establishment of dangerous weeds. This study includes samples of ~ 200 species of Stipoid grasses (Stipeae, Poaceae). After trialling various DNA markers, four regions of DNA were sequenced including the nuclear internal transcribed spacers (ITS), trn Q-5’ rps 16 intergenic spacer, ribulose bisphosphate carboxylase/oxygenase large subunit ( rbc L) and maturase K ( mat K). The resulting sequences, having been obtained from expertly determined and vouchered specimens, were used to assemble a comprehensive reference library of known sequences of Stipoid grasses, against which unknown sequences could be compared.
We provide comparison of the resolving power of the three different DNA regions, both individually and in combination, to provide identification of any particular species in this group of invasive grasses and their relatives. Critically, we also compare different methods of sequence analysis, for matching unknown samples to the database of known samples, to ascertain the most effective method for maximal discrimination of species.
On this basis recommendations will be made for the most effective molecular identification of Stipoid grasses, leading to potential savings in time and money, and increased accuracy when dealing with these biosecurity threats.
Tree DNA barcoding of the Australian Wet Tropics: species delineation and diversity estimation
Costion, C 1, 2 , Crayn, D 1 , Lowe, A 2
1
2
Australia Tropical Herbarium, Cairns, Australia;
University of Adelaide, Australia
DNA barcoding, the use of short, standardised DNA sequences for species identification is gaining much popularity and publicity for its promise in the near future to enable any biological material containing DNA to be accurately identified. The applications and implications of this technology are broad, from CITES regulation to classroom biology for teachers and students. Although the selection of a universal DNA barcoding region for plants has been controversial, two genes: rbc L and mat K have been recently selected. Our project presents a combined rbc L & mat K dataset of over 500 species of
Queensland Wet Tropics plants, each represented by at least three separate collections. This is the largest of its kind for Australian tropical plants to date. We test the accuracy of these markers for species delineation in
Australia and are investigated the relative robustness of
DNA barcodes to estimate the number of species of a given area assuming the species are not known. We tested this by sampling all trees (>10cm dbh) in plots with a known species composition. Our results provide insight into the exciting potentials and the limitations that need to be addressed for DNA barcode data to be utilized as a rapid species inventory and diversity assessment method.
Plant DNA barcoding using chloroplast genome sequences
Nock, C 1 , Waters, D 1 , Shepherd, M
1
Henry, R 2
1 , Bundock, P 1 ,
Centre for Plant Conservation Genetics, Southern Cross
University, Australia; 2 Queensland Alliance for
238
Agriculture and Innovation, University of Queensland,
Australia
Chloroplast DNA sequence data have played a critical role in the development of plant DNA barcodes. While the mitochondrial locus CO1 is well accepted as an efficient DNA barcode for animals, no single locus has been identified that can discriminate between all plant species. There has been considerable debate about the selection of the most suitable chloroplast loci, and the two-locus ( mat K and rbc L) plant barcode proposed in
2009 remains problematic for some plant groups. Recent advances in DNA sequencing technology are providing opportunities for whole genome sequence comparisons.
Assembly of chloroplast genome sequences from massively parallel sequencing (MPS) of total DNA circumvents PCR-based issues such as amplification and primer universality. Access to the whole chloroplast genome will provide many more informative sites and has the potential to improve plant identification within and among even closely related species. We present chloroplast genome sequences recovered from MPS of total DNA for a wide range of species including wild rice relatives, eucalypts and sugarcane. The analysis of chloroplast genome sequences is rapidly becoming a simple, accurate and cost effective option for DNA barcoding and plant identification. Is the chloroplast genome the elusive single-locus DNA barcode for plants?
Utilizing grass DNA barcodes to explore Australian ecosystems
1
Cross, H 1, 2 , Murphy, D
A 3 , Jardine, D
3 , Brown, A
2 , Cantrill, D
2
3 , Lowe, A
, Gardner, S 3
1, 2
, Syme,
State Herbarium of South Australia, Australia;
2 Australian Centre for Evolutionary Biology and
Biodiversity (ACEBB), The University of Adelaide,
3 Australia; National Herbarium of Victoria, Royal
Botanic Gardens Melbourne, Australia
The grasses (Poaceae) are one of the largest families of plants in the world. They contain our most important grain crops, and as well as many of the world’s most problematic invasive species. In addition, grasses form a critical component of many ecosystems and range from the poles to the tropics, in extremes of heat and aridity.
The Grass Barcoding of Life project (GrassBoL) aims to unite plant researchers from around the world in DNA barcoding all species of the order Poales, extending from the grass family to include taxa with similar form and habitat to the true grasses. In Australia we have obtained
DNA barcodes for all of the important groups of grasses.
While our taxon sampling has been broad, we have also focused on select groups to test the ability of DNA barcodes to distinguish among closely related genera and species. Our initial results have found that much work is needed to clarify the taxonomy and species relationships of many groups. Due to the broad ranging economic and ecological importance of grasses, the results of the
GrassBoL project are being integrated into several other research initiatives in Australia and overseas. These include major projects on ecosystem monitoring, biosecurity protection, cereal crop genomics, dietary studies of outback fauna, and even ancient DNA studies of paleosoils. We report on the efficacy of DNA barcodes

in grasses but also how they are being utilized in other research projects by providing baseline genetic information and reference DNA sequences.
The global two-locus plant barcode: a first evaluation of its utility across ferns
Li, F-W 1 , Rothfels, C 1 , Windham, M 1 , Pryer, K 1
1 Dept of Biology, Duke University, USA
Recent approval of the plastid loci rbc L and mat K as the global DNA barcode for plants posed a serious challenge to systematists working on ferns (the second largest clade of vascular plants). In most plants, mat K is nested within a trn K intron in the large single copy region of the chloroplast and is easily sequenced. In ferns, however, the stable priming sites in the flanking trn K exons were lost, hindering an evaluation of mat K in this lineage.
Recently, new primer design has helped to overcome this obstacle and mat K sequences are now available for representatives from all fern families. Here we provide a first assessment (and endorsement) of the utility of rbc L and mat K as a barcode across ferns. In addition, we present data for a third plastid locus, trn Hpsb A, that is now in strong contention to be added to the global plant
DNA barcode because of its high substitution rate in angiosperms. In ferns, however, we find the average sequence divergence of trn Hpsb A to be extremely low
(even lower than rbc L). This is likely due to a striking genome rearrangement early in fern evolution that resulted in its inclusion in the inverted repeat (IR) region.
Our more detailed examination of trn Hpsb A within a genus of leptosporangiate ferns also reveals a mononucleotide repeat whose length varies at both the inter- and intra-specific levels (individuals of different species can have the same length of the repeat, whereas it is sometimes inconsistent within a species). This highly dynamic repeat length, together with its lower molecular rate of evolution, will undoubtedly diminish the candidacy of trn Hpsb A as a recruit for DNA barcoding in plants.
DECLIC: an analytical pipeline for delimitation of species using cliques and its application to molecularbased biodiversity inventory
Franc, A 1 , Chaumeil, P 1 , Frigerio, J-M 1
1
Molino, J-F 2
Saintagne, C 3
, Léger, P 1
, Petit, RJ 1
, Caron, H 1
, Sabatier, D 2 , Scotti, I 3 , Scotti-
,
INRA – BioGeCo, Bordeaux, France;
Montpellier, France;
Guiana, France
3
2 IRD – AMAP,
INRA – Ecofog, Kourou, French
Most ecological studies, especially in tropical rainforests, require a comprehensive botanical inventory. Inventories usually exist for long term research plots, but are less common for new plots, or in poorly known floras.
Molecular taxonomy can be used to estimate species diversity through 'blasting' community sequence data against established databases (e.g. NCBI, BOLD) but this is time demanding. Here, we present a pipeline, called
DECLIC, which, describes data structure based on similarity in genetic distances for large sequence datasets from inventory studies. It improves on standard methods which generate p-distances from global alignments by generating a pairwise global edit distance matrix, avoiding the need for a global alignment. Thus it is particularly useful for datasets which are large, taxonomically diverse, and highly variable (e.g. trn Hpsb A) and multiple alignment is not feasible. DECLIC allows the user to work with datasets up to a few thousands items, and soon a few tens of thousands. Much effort has been invested in computing efficiency for analyzing large data sets within reasonable time (a few hours at most). This pipeline can be used as a tool for examining if, and how, molecular variability in a set of markers on a set of individuals, is related to taxonomic hierarchy as assessed by botanists. While some population genetic studies suggest that taxa may not be real (because of widespread introgression, for example), initial results using DECLIC show that molecular variability mostly is organized into clusters and isolated items. The method will be presented, namely how the distances are translated into a graph, how connex components and cliques are retrieved automatically from the graph. Some results will be shown, especially on a large data set (1733 individuals from about 500 species,
220 genera and 35 families, representing a comprehensive sample of the French Guianan tree flora), where it is shown that well defined components of the graph, be their connex components or cliques, usually match recognized taxonomic units, mostly at genus or subgenus level on plastids. Other results will be shown, as comparisons with molecular phylogenies on smaller data sets, showing that the pipeline retrieves well supported clades in molecular phylogenies, which is another argument for its validation. Finally, the advantages and drawbacks of BLAST, phylogenies and
DECLIC will be discussed for large scale inventories or surveys.
Is DNA barcoding child’s play? Using citizen scientists to test the utility of plant barcoding
Tosh, J 1 , Crookshank, A 2 , Hopkins, D 1
1 Natural History Museum, London, UK;
Malthouse, Dorset, UK
2 The Old
Since 2009, scientists from the Natural History Museum,
London, have been working with the Cothill Educational
Trust to look at innovative ways of educating and enthusing school children in the natural sciences. ‘Tree
School’ is the first project resulting from this joint venture, and provides children (aged 10 and upwards) with the opportunity to learn about tree identification, biodiversity and DNA barcoding during weeklong residential workshops at the Old Malthouse on the Isle of
Purbeck (Dorset, UK). The children select a tree for the week that they identify, voucher and DNA barcode. Their
PCR products are then sequenced and assembled by scientists at the Natural History Museum. Here we present the results of all the children’s data generated to date, assess the utility of rbc L and mat K as plant barcodes in the context of a floristic study and discuss the merits of ‘Tree School’ from the perspective of both scientists and educators.
239

Progress toward a comprehensive molecular phylogeny of legumes: are we almost there?
Wojciechowski, M 1 , Sanderson, MJ 2 , Herendeen, P 3
1 Arizona State University, USA;
USA; 3
2 University of Arizona,
Chicago Botanic Garden, USA
We have undertaken a concerted effort to reconstruct a more comprehensive phylogeny of legumes
(Leguminosae) by utilizing the wealth of molecular sequence data publically available in Genbank databases.
Using recently developed methods for parsing and processing potentially informative sequences in these databases, we have constructed maximal clusters of two plastid genes, matK and rbcL, both widely used for previous higher-level phylogenetic analyses of the family, to serve as a framework for the identification and assembly of additional clusters of both coding and noncoding sequences (e.g., introns, spacers) from the same set of taxa into a concatenated, multigene supermatrix for subsequent analyses. Clusters of these two genes include a minimum of 1100 and 665 taxa, respectively, and together include more than 400 of the c. 740 genera which combined represent more than 80% of the taxonomic diversity (species-level) in the family. The results of our phylogenetic and fossil-calibrated molecular dating analyses and their general implications greatly refine our understanding of the evolutionary history, and will ultimately lead to a more robust and stable higher-level classification, of this angiosperm clade.
The Caesalpinioideae: purging polyphyly and pacifying paraphyly to clarify classification
Mackinder, BA 1
GP 1
, Bruneau, A 2 , Herendeen, PS 3 , Lewis,
1 Royal Botanic Gardens Kew, UK;
Montréal, Canada; 3
2 Université de
Chicago Botanic Garden, USA
Leguminosae comprises three subfamilies;
Caesalpinioideae, Mimosoideae and Papilionoideae. The former is composed of c. 2300 species, in 171 genera which are currently recognized in four tribes. During the last decade, numerous small to large scale phylogenetic studies incorporating nucleotide and morphological data have substantially improved our understanding of generic and supra-generic relationships in Caesalpinioideae. Of the four tribes presently recognised, we now know that
Cercideae and Detarieae are monophyletic while
Caesalpinieae and Cassieae are polyphyletic. Within all four tribes several well-supported supra-generic groups have been resolved as monophyletic such as the
Detarieae s.s., Amherstieae, Cassia and Caesalpinia clades although strong internal resolution is sometimes lacking. Many caesalpinioid genera for which multiple species have been sampled are robustly supported as monophyletic including some of the most species rich such as Chamaecrista and Senna of tribe Cassieae.
240
However, other genera including some large and complex genera such as Caesalpinia s.l. and Bauhinia s.l.
(Cercideae) are not monophyletic and their future classification will likely require some recircumscription of generic boundaries. We have established that
Cercideae and Detarieae are two of the first branches of the legume phylogeny. Other early branching caesalpinioid lineages are those of the Dialiinae, recently defined to include two subtribes of the Cassieae and the genus Poepiggia and a fourth lineage which consists only of the monotypic genus Duparquetia . Thereafter follows the rest of the legumes, placed in two major clades, the
Papilionoideae clade and a clade that contains the
Mimosoideae, the Umtiza Clade, some genera of the
Cassieae and assorted Caesalpinieae lineages. Within the latter major clade, is a subclade that contains some of the genera of the Dimorphandra group (Caesalpinieae) placed as a paraphyletic grade at the base of the
Mimosoideae. Consequently, in a phylogenetic classification of the legume family the caesalpinioids would not be recognized as a single subfamily.
Nevertheless, we can delineate many strongly supported major and minor clades of caesalpinioid legumes and have a clear view of how they relate to each other.
Problems and possibilities: alternative classifications for the Mimosoid legumes
Luckow, M 1
Miller, J 5
, Hughes, C
, De Quieroz, L 6
2 , Simon, M
, Lewis, G 7
3 , Grether, R 4 ,
1 Cornell University, New York, USA;
Systematic Botany, Zürich, Switzerland;
2 Institute of
3 Embrapa
4
Recursos Genéticos e Biotecnologia, Brasília, Brazil;
Universidad Autónoma Metropolitana-Iztapalapa,
5 Mexico; Centre for Plant Biodiversity Research,
Canberra, Australia; 6 Departamento de Ciências
Biológicas, Universidade Estadual de Feira de Santana,
Feira de Santana, Brazil; 7 Royal Botanic Gardens, Kew,
UK
Since 1982, five tribes have been frequently recognized in the Mimosoideae: Parkieae, Mimozygantheae,
Mimoseae, Acacieae, and Ingeae. Molecular phylogenetic studies over the past decade showed that most of these tribes were either para- or polyphyletic.
Generic rearrangements were proposed in the 2005 book
Legumes of the World resulting in the recognition of only four tribes (the Parkieae was dismantled). Although the new system resulted in a monophyletic Acacieae and
Ingeae, the Mimoseae remain a paraphyletic grade at the base of the mimosoids, partitioned into informal groups.
It is now clear that tribe Mimozygantheae should also be included in Mimoseae. Our work will address the question of how or whether to recognize tribes in the
Mimosoideae, and what groups would be most useful and logical given both recent molecular phylogenies and the distribution of morphological characters on those trees.
The need for additional molecular data to resolve particular nodes is discussed, and the recircumscription of the subfamily is also touched upon in the context of all legumes.

Progress in Papilionoid legume systematics: towards a phylogenetic classification
Boatwright, S 2 , Lavin, M 3 , McMahon, M 4
Sanderson, M 4
, Kenicer, G
, Steele, K 5
Wojciechowski, MF 5
1
, Torke, B 6 , van Wyk, B-E 7
,
,
5
1 Royal Botanic Garden Edinburgh, UK;
University of Arizona, USA;
Arizona State University, USA;
Garden, USA; 7
4
2 South African
National Biodiversity Institute, South Africa;
State University, USA;
6
3 Montana
New York Botanical
University of Johannesburg, South Africa
The Papilionoideae, as the largest and most wonderful subfamily of the Leguminosae (478 genera, 13,800 species), presents considerable challenges for systematic research. Phylogenetic studies using principally chloroplast gene and intron sequences have delimited a series of major clades in the subfamily. However, in some cases relationships amongst these groups is not clear, taxon sampling within them has not been adequate, and there is a need to integrate morphological characters.
This presentation will review the implications for
Papilionoid systematics of a new family-wide molecular phylogenetic analysis of legumes produced by the
Legume Phylogeny Working Group. It also draws on recent phylogenetic studies of clades within the subfamily. It aims to focus priorities for future research that can lead to a stable phylogenetically-based classification for Papilionoideae.
Metacommunity ecology rather than unique historical events have shaped the global biodiversity of the
Leguminosae
Lavin, M
G 3
1 , Pennington, T
, Hughes, C 5
2 , Wojciechowski, M 4
, Delgado, A 6
, Lewis,
, Schrire, B 3 , Thulin, M 7
1 Montana State University, USA;
Gardens, Edinburgh, UK;
UK; 4 of Zurich, Switzerland;
Mexico City, Mexico;
6
7
3
2 Royal Botanic
Royal Botanic Gardens, Kew,
Arizona State University, Tempe, USA; 5 University
National University of Mexico,
Uppsala University, Uppsala,
Sweden
Molecular phylogenetic studies of transcontinental legume clades reveal no consistent geographical patterns and often more ecological than geographical structure.
Furthermore, evolutionary substitution rate analyses that bias for fast substitution rates and thus young ages consistently find ages of transcontinental crown clades to be often much less than 30 Ma. This contrasts to the ages of hypothesized continental tectonic events, most of which are older than 30 Ma, which may have facilitated plant migration among continents. The conclusion is that legumes disperse and immigrate very well without the aid of land bridges and stepping stones. What constrains their immigration appears to be more a function of ecology than geography. As a consequence, community rather than taxon phylogenetics may provide the most promising avenue for identifying the important ecological parameters that shape legume biodiversity.
Leguminosae – how stable are the supra-generic
1 clades and are we ready for a new classification?
Lewis, G 1
Royal Botanic Gardens, Kew, UK
The two volume proceedings of the first international legume conference ( Advances in Legume Systematics parts 1 & 2) published in 1981 contained no evolutionary trees or phylogenetic analyses. When Advances in
Legume Systematics part 7 (subtitled 'Phylogeny') was published in 1995 nearly every paper included at least one phylogenetic tree as an integral component, thus marking the enormous advance of the science in little over a decade. From 1995 to present there has been a plethora of phylogenetic analyses published at all hierarchical levels, from species to subfamily. Data for such analyses has emerged from a wide range of disciplines, including anatomy, genome sequences, morphology, phytochemistry, and combined data sets.
Results have shown that Caesalpinioideae are not monophyletic, and a large number of former tribes, subtribes and large genera throughout the legumes are likewise unsupported. In Legumes of the World (2005), the recognised 727 genera were organised systematically in an arrangement underpinned by the latest phylogenetic information. Although this represented a major step towards a new classification of the family most suprageneric clades had not been comprehensively sampled in phylogenetic analyses and were therefore not given formal nomenclatural status. Since 2005 the number of accepted legume genera has increased to 738, and a burgeoning phylogenetic literature continues to focus on resolving relationships within and between the major legume clades. The 36 legume tribes in Legumes of the
World are likely to expand to at least 54 in any new classification, and the three former subfamilies will become eight or nine. In 2010 the international Legume
Phylogeny Working Group (LPWG) was set up with the aim to combine all data from the three traditionally recognised subfamilies: Caesalpinioideae, Mimosoideae and Papilionoideae and undertake a complete family analysis. Its goal would be to deliver a well-supported global legume phylogeny. A list of genera exists. What is required is a critical species list for the family so that all researchers can agree on (1) sampling from the same specimens ('gold star') or at least the same species ('silver star') where adequate material exists for multidisciplinary study, (2) key morphological characters that specialists should score in all three subfamilies and (3) the gene loci that should be selected for molecular studies. This approach should result in a cross-family, combined-data phylogenetic analysis that gives us a robust family tree which can be used as a basis for a new consensus classification. The talk will synthesize data put forward in the other five presentations of the session, assess the stability of the current major supra-generic legume clades and consider whether formal nomenclatural recognition is advisable at this stage.
Leaf anatomy of 58 species of
Chamaecrista
Moench. as an additional taxonomy tool to the molecular phylogeny
Coutinho, I 1 , Francino, D 1 , Meira, R 1
241

1 Universidade Federal de Viçosa, Brazil
Chamaecrista Moench., one of the largest genera of
Leguminosae–Caesalpinioideae (circa 330 spp.), has the
East and Midwest areas of Brazil as the area core of diversity. The genus is divided into six sections:
Apoucouita , Absus , Grimaldia , Chamaecrista ,
Caliciopsis and Xerocalyx . The presence of glandular trichomes is one of the characters that delineates the sections Absus (165 spp.) and Grimaldia (1 spp.). The species of the section Absus are concentrated at the
Rupestrian Fields (Rocky Outcrops) and Cerrados
(Brazilian Savannah). Absus is subdivided into four subsections: Adenophyllum , Baseophyllum , Otophyllum and Absus . The subsection Absus has around 160 species which are sorted out into 31 series, a classification poorly supported by morphological characters. Molecular studies support the genus Chamaecrista as a monophyletic group; however, approaches concerning the sister group and infrageneric relationships are unsatisfactory. Anatomical studies have been recently used as an additional tool to the molecular phylogeny.
The present study aims to anatomically characterise the leaves of 58 species of Chamaecrista as well as to evaluate the importance of the anatomical characters as subsides to the taxonomy and phylogeny of this genus.
Samples from voucher material and plants collected in the field were processed according to ordinary plant anatomy techniques for the structural characterisation. A presence/absence matrix based only on anatomical characters was built. Principal components analysis
(PCA) and cluster analysis were calculated. The phenogram was built using the UPGMA (Unweighted
Pair Group Method with Arithmetic Mean). The following xeromorphic characters were common among the Chamaecrista species studied: isobilateral leaves, compact mesophyll, pubescence, high vein density, abundance of mechanical tissue, and epidermal cells with thick outer periclinal walls. The group analysis resulted in a phenogram where two main groups where formed.
The first largest group has species bearing extrafloral nectaries (sections Chamaecrista and Xerocalyx , and section Absus subsections Baseophyllum and
Otophyllum ). Species belonging to the subsection
Baseophyllum formed a subgroup supported by two main characters: epistomatic leaves and mucilaginous cells present in the mesophyll. Species from the section Absus subsection Absus and the species from the monotypic section Grimaldia form the second largest group, which is characterised mainly for the presence of secretory trichomes and mucilaginous epidermis. The anatomical characters that stand out on the outlining of the subgroups are: presence of trichomes on reproductive and/or vegetative organs, distribution of secretory trichomes on the leaf blade, secretory trichomes with a very short stalk, a large and spherical shaped head vs. secretory trichomes with a slender head both bearing a filiform projection, and prominence of the midvein. Our finding corroborate with the molecular phylogeny which suggests the elevation of subsection Baseophyllum to a section status, and the relocation of Chamaecrista absus
(the single species in the Grimaldia section) to the section Absus . Also, the sorting of the subsection Absus into 31 series is merely artificial. Hence, additional studies taking into account taxonomic research reviews and both molecular and morphological phylogenies for a better outlining and understanding of the relationships at
242 the infrageneric level. We thank CNPq and FAPEMIG for the financial support.
Structural diversity of the wood of
Acacia
in
Australia in relation to phylogeny and climate
Gasson, P 1 , Warwick, N 2 , Whinder, F 1 , Hailey, L 1
1 Royal Botanic Gardens Kew;
England, Armidale, Australia
2 University of New
There are more than 1000 species of Acacia in Australia spread across a wide range of climatic zones – arid, semiarid, temperate, alpine, sub-tropical and tropical. They are diverse in floral and leaf morphology, most strikingly between the bipinnate section Botrycephalae and the phyllodinous sections Juliflorae , Phyllodineae and
Plurinerves . Phylogenetic studies by Miller et al. indicate that these section groupings may not be entirely appropriate. This study investigates the structural diversity of the secondary xylem of Acacia in the context of the four sections and in four climate zones. We have collected and examined woody stem samples from approximately forty species from natural populations in arid, semi-arid, temperate and sub-tropical zones. The material was fixed in 70% ethanol and sectioned in three planes (transverse, tangential and radial) for light microscopy. Some specimens were prepared for SEM examination. The wood is generally dense, with thickwalled vessels and fibres, paratracheal parenchyma and narrow rays 1–2 cells wide. Growth rings are often well defined and vestures are found in all the intervessel and vessel-ray pitting. Prismatic crystals are often abundant in chambered axial parenchyma and fibres. Our observations will also be discussed in the context of the physiological requirements of acacias in the different environments.
Phylogeny of tribe Ingeae – the forgotten legumes
, Murphy, D 2 , Ladiges, P 1
1
Brown, G 1
School of Botany, The University of Melbourne,
Australia; 2 National Herbarium of Victoria, Royal
Botanic Gardens Melbourne, Australia
Tribe Ingeae is a significant group of legumes, characterised by the possession of valvate calyx-lobes and numerous stamens fused into a tube. The classification history of this large tribe, which consists of c. 1000 species in 36 genera, has been complex and confused due to the large morphological variation, vast geographic distribution and relatively limited collection of herbarium samples with taxonomically informative characteristics (i.e. fruiting specimens). Ingioid taxa occur throughout tropical regions, with two main centres of diversity: one in Central and South America, and a second in southeast Asia and Australia. Tribes Acacieae and Ingeae are unquestionably linked, with tribe Ingeae nested within tribe Acacieae, and a revision of their tribal boundaries is required. However, very little is known about the phylogenetic relationships of taxa within tribe
Ingeae, in comparison to Acacieae. There is currently no well supported phylogeny (hence inter-generic classification) of the tribe Ingeae despite its size, confused taxonomy and importance in tropical ecosystems. Several molecular phylogenies have

included taxa from the Ingeae and results suggest that some large genera (e.g. Albizia and Archidendron ) are paraphyletic, although, relationships between clades are not well resolved and many genera remain to be sampled.
This study aims to close this knowledge gap by investigating the relationship of taxa within tribe Ingeae and related clades of Acacia s.l., and by exploring the monophyly of genera and informal alliances using nuclear ribosomal DNA and chloroplast DNA sequences.
Interpreting morphological complexity and parallelism within the framework of a phylogenetically-based infrageneric classification of
Swartzia
(Leguminosae)
Torke, BM 1 , De Freitas Mansano, V 2
Bohórquez3, L 3 , Campbell, L 4
, Karen Ruiz
1 Institute of Systematic Botany, New York Botanical
Garden, USA; 2 Instituto de Pesquisas, Jardim Botânico
3 do Rio de Janeiro, DIPEO, Brazil; Museo de Historia
Natural ANDES, Depto de Ciencias Biológicas,
Universidad de los Andes, Colombia; 4 Pfizer Plant
Research Laboratory, New York Botanical Garden, USA
Recent improvement in understanding of phylogenetic relationships within the species-rich neotropical tree genus Swartzia (Leguminosae) forms the basis of a new infrageneric classification and also sheds light on longstanding questions about the morphological evolution of the genus. While many of the features that characterize multiple clades of Swartzia , including an entire calyx lacking regular sepals, a single petal, a zygomorphic androecium with dimorphic stamens, and arillate seeds, are ancestral in the genus; other, less common but equally widespread features, such as apetaly, carpel duplication, isomorphic stamens, and stalked trichomes, have arisen in parallel numerous times during the evolutionary history of the genus. We present preliminary evidence to suggest that morphological parallelism is linked to a lack of morphological canalization and/or developmental stability and a history of ecological parallelism.
A molecular phylogenetic analysis of invasive and ornamental brooms and their relationships within the genistoid legumes
Kleist, A 1 , Jasieniuk, M 1
1 University of California – Davis, USA
The Cytisus – Genista complex has a center of diversity in the Mediterranean region, but several members of the group have become invasive weeds in many parts of the world following introduction as ornamental plants.
Despite their widespread occurrence and impact, the evolutionary relationships among invasives, ornamentals and native-range species have not been studied using molecular tools. Closely related species and varietals are still available commercially, so understanding the relationships and evolution of the group is vital for biocontrol efforts and preventing future invasions. For this study, we focused specifically on invasive French broom in western North America because it inhabits a wider variety of habitats in this invaded range than the native range and its taxonomic identity was unresolved.
Although commonly considered to be Genista monspessulana , invasive French broom has also been suggested to be a hybrid between G. canariensis and G. stenopetala or a hybrid between G. monspessulana and a currently available ornamental plant, sweet broom. The objectives of this project were to: i) determine the taxonomic identity of invasive French broom and closely related ornamental brooms; ii) reconstruct the phylogenetic relationships of invasive and ornamental brooms to each other and to the Genistoid legumes as a whole; and iii) assess whether hybridization between ornamental cultivars, species, and naturalized populations has occurred. To address these goals, we collected samples from landscape plantings, horticultural outlets, and invasive French broom populations from a range of habitats throughout California, and from botanical gardens and arboreta from the native range. A combination of maternally and biparentally inherited plastid and nuclear DNA sequence and microsatellite data were used to investigate the origins and evolutionary relationships of invasive French broom. Our phylogenetic analyses recovered separate French broom and sweet broom groups within the monspessulana clade in the genus Genista . Additionally, cloning and sequencing of the ITS nuclear DNA region revealed that although the majority of invasive French broom in
California is G. monspessulana , hybridization with individuals from the ornamental sweet broom clade occurs in populations throughout the state of California.
We are currently using microsatellite markers, which have substantial variability and co-dominant inheritance, to further investigate hybrid origin and parentage.
1
Hydatellaceae and early angiosperm evolution
Rudall, P 1
Royal Botanic Gardens Kew, UK
The traditional paradigm of early angiosperm evolution as an incremental accumulation of adaptive innovations leading to the major diversifications of monocots and eudicots is contradicted by the high degree of morphological diversity among early-divergent extant angiosperms. The problem is compounded by large numbers of ancient extinctions. The recent robust phylogenetic placement of Hydatellaceae as sister to the waterlilies places this family among the most ancient surviving lineages of angiosperms. This remarkable discovery represents a unique opportunity for comparative studies of early angiosperm evolution. Like
Hydatellaceae (1 genus, 12 species), most extant earlydivergent angiosperm lineages are relatively speciespoor, but assume disproportional significance in comparative studies of angiosperm evolution. Species of
Trithuria (Hydatellaceae) are inconspicuous plants, mostly from the Australian subcontinent, but with outliers in India and New Zealand. They are interesting not only for their critical phylogenetic placement and highly unusual morphology, but also for their disjunct biogeography, diversity of life strategies and ecology in seasonal wetland habitats. Detailed comparative studies
243

are establishing these plants as emerging new model organisms among early-divergent angiosperms.
Phylogenetics and evolution of Hydatellaceae
Iles, WJD 1
MV 3
SW 1
, Rudall, PJ 2
, Lee, C 1
, Sokoloff, DD
, Barker, MS 4
1 University of British Columbia, Canada; 2
3
3 , Memizowa,
, Rieseberg, LH 1 , Graham,
Royal Botanic
4
Gardens Kew, UK; Moscow State University, Russia;
University of Arizona, USA
The small Australasian family Hydatellaceae was considered to be closely related to Centrolepidaceae in the commelinid monocots until recently. This taxonomic mis-association reflects a superficial resemblances in habit (both are tiny aquatic herbs) and floral morphology
(pseudanthia-like reproductive units). However, it is now recognized that Hydatellaceae are related to the water lilies, Cabombaceae and Nymphaeaceae. We inferred phylogenetic relationships among the 12 recognized species based on multiple plastid regions. We used this as a framework for examining species delimitation, estimating clade divergence times, and for reconstructing the evolution of morphological characters, sexual systems and abiotic niche preferences. We recovered four major clades that can be partly delineated using fruit and seed characteristics. We resolved initial uncertainty about the placement of the root node of the family by sampling a a subset of taxa for a larger set of plastid genes, confirming that a primarily northern Australian clade is the sister group of the remainder of the family. Our species-level phylogenetic survey included multiple populations for most species, and we were able to resolve most interspecific relationships, generally with good support. However, support for the monophyly of several species was lacking, and one species was strongly supported as non-monophyletic. We also attempted to date the origin of Hydatellaceae using relaxed clock methods and a variety of fossil constraints. We estimate that the family diverged from the line leading to the water lilies at least 100 million years ago. However, accurate dating of the crown age of the family is confounded by extensive rate elevation and a dearth of fossil evidence. Nonetheless, we infer relatively recent long-distance dispersal events from Australia to India and New Zealand. Our species-level phylogeny also implies that there was substantial lability in sexuality in the family, with frequent switching between dioecy and cosexuality. Switches in sexual system appear to be correlated with morphological changes associated with male reproductive output. The climatic breadth of the family is relatively large. We explored abiotic niche evolution by inferring ancestral values for relevant abiotic variables, and consider the possible interplay between inferred ancestral climatic niches and paleoclimates. We also report on efforts to use transcriptome data sets from Hydatellaceae and other flowering plants to pinpoint early episodes of whole genome duplication in angiosperm phylogeny.
Plant architecture and shoot branching patterns in early-divergent aquatic angiosperms, with emphasis on Hydatellaceae
Sokoloff, DD 1
Tuckett, RE 4
, Remizowa, MV 1 , Beer, AS 1
Sadovnikova, ES 1 , Yadav, SR 2
, Briggs, BG 5
, Macfarlane, TD
, Rudall, PJ 6
3
,
,
1 Faculty of Biology, Moscow State University, Moscow,
Russia; 2 Shivaji University, Kolhapur, India; 3 Western
Australian Herbarium, Dept of Environment &
Conservation, WA, Australia;
Australia, Crawley, Australia;
Sydney, Australia; 6
4 The University of Western
5 Royal Botanic Gardens,
Royal Botanic Gardens, Kew, UK
Both molecular phylogenies and the fossil record show that aquatics merit special attention for understanding early angiosperm evolution. Two of six extant clades of early-divergent angiosperms consist of aquatics: the monogeneric Ceratophyllaceae and the order
Nymphaeales (Cabombaceae, Hydatellaceae,
Nymphaeaceae). The period of early angiosperm diversification coincided with that of extensive diversification of freshwater aquatic plants, including non-angiosperm groups. Although some Lower
Cretaceous angiosperm aquatics are considered closely related to either Ceratophyllaceae or Nymphaeales, it is likely that other (now extinct) aquatic lineages also existed among the earliest angiosperms. Axillary branching is a common feature of most seed plants, but many extant early-divergent angiosperm aquatics diverge from this pattern. Flower arrangement is an important aspect of shoot branching patterns in many angiosperm groups, but recognizing flowers within the plant body can be problematic, at least in Hydatellaceae,
Ceratophyllaceae and the aquatic fossil Archaefructus .
The present study compares shoot architecture in
Nymphaeaceae and Hydatellaceae, and examines whether deviations from axillary branching patterns result from reduction of subtending phyllomes. Despite many important observations on shoot branching and flower arrangement in Nymphaeaceae since the nineteenth century, morphological and evolutionary interpretations remain controversial. In Victoria and
Euryale , flowers are axillary on the unbranched shoots, but are not inserted in the centre of the leaf axil. In
Nymphaea and Nuphar , flowers appear to occupy some sites of the phyllotaxy spiral along the monopodial rhizome. One possible interpretation of this arrangement is that a primordium initiated by the shoot apex can develop either as a foliage leaf or as a flower. An alternative interpretation is that the flower-subtending bracts are suppressed. In Nuphar , a vestigial phyllome is sometimes visible at the abaxial side of the pedicel base; this could represent a reduced flower-subtending bract, though reports of two phyllomes associated with the pedicel make this interpretation problematic. A third possibility, of morphologically terminal flower insertion and sympodial rhizome construction, should be also considered. In contrast with Nymphaeaceae, shoot architecture was little known in Hydatellaceae until a recent study of the perennial species Trithuria filamentosa . Rhizomes of this species are sympodial.
Stalks bearing reproductive units can be interpreted as the uppermost lateral extra-axillary organs that continue the shoot phyllotaxic spiral; in this hypothesis, the mode of insertion of reproductive units in T.
filamentosa
244

resembles that of the flower in Nymphaea . Annual species of Hydatellaceae apparently possess more than one model of shoot architecture. In Trithuria australis , vegetative leaves occur only on the main shoot, as in
Victoria . A male reproductive unit appears to be terminal while female units are lateral. Subtending leaves of female units are absent, and the units appear to initiate at least partially basipetally. In Trithuria konkanensis , shoots are branching, and foliage leaves are present on the shoots of different orders. An axillary position cannot be demonstrated for most reproductive units in this species.
Reproductive life history variation and breeding systems in
Trithuria
(Hydatellaceae)
Taylor, ML 1 , Williams, JH 1
1 University of Tennessee, Knoxville, TN, USA
With the origin of angiosperms, the reproductive cycle became much shortened, particularly during the progamic phase (the period between pollination and fertilization). Nymphaeales (water lilies) is an ancient lineage of flowering plants, arising from the basal-most or next most basal node of the angiosperm phylogenetic tree. Within Nymphaeales, Trithuria may represent an extreme in progamic phase compression because of its reduced size and shorter overall life history – it is one of the few early-diverging angiosperm lineages to evolve annual life histories. Trithuria species also exhibit remarkable diversity in sexual system and habitat, which may also affect their fertilization biology. We conducted field studies of Trithuria submersa , T. australis , and T. austinensis in southwestern Australia. These species are bisexual, monoecious, and dioecious, respectively, and inhabit a range of aquatic or semi-aquatic habitats. Handpollinations and experimental manipulations were performed over two field seasons to assess the timing of reproductive events such as stigma receptivity, pollen reception, and pollen tube growth. Breeding system characteristics, including primary pollen vector, pollen/ovule ratios and the potential for self-fertilization were also determined. All three Trithuria species exhibited some degree of wind pollination. Pollen tube pathway lengths ranged from 539 µm in T. submersa to
2262 µm in T. austinensis . However, all species exhibited rapid pollen germination and relatively fast pollen tube growth rates. A short pollen tube pathway and rapid pollen tube development result in an extremely brief progamic phase that, in T. submersa , ranks as one of the shortest known among all angiosperms. Trithuria submersa and T. austinensis represent two extremes of reproductive strategies. T. submersa is rarely crosspollinated (mean outcross-pollen loads < 0.5 grains/ovule), but is effectively self-pollinated and produces high self-seed set. These facts and its low pollen/ovule ratio (24), indicate that T. submersa has a primarily autogamous breeding system. T. submersa commonly inhabits small disturbed wetlands in which heterogeneous microsites cause non-synchronous plant emergence. Selfing, along with rapid reproduction and small plant size, likely enables T. submersa to colonize such ephemeral habitats. Trithuria austinensis , in contrast, is dioecious and obligately outcrossing. This species has a pollen/ovule ratio of 1569 and more efficient out-cross pollen transfer – it has a mean pollen load almost 5 times that of T. submersa . As might be expected, this species grows at much higher population densities and its stigmas have a greater receptive surface than those of T. submersa . T. austinensis is found in environments such as large shallow basins, where habitat homogeneity promotes synchronous plant emergence and effective cross-pollination. Our preliminary data indicate that monoecious T. australis has a low pollen/ovule ratio and exhibits selfing. Trithuria species have evolved very divergent breeding systems, from autogamy to dioecy.
Breeding systems in Trithuria are intimately tied to reproductive life history and habitat and are reflected in reproductive morphology. Trithuria is an ancient genus that displays great evolutionary lability within a number of marginal habitats.
Biology of
Trithuria konkanensis
, the sole Indian species of Hydatellaceae
Yadav, SR
PJ 3
1 , Sokoloff, DD 2 , Remizowa, MV 2 , Rudall,
1 Shivaji University, Kolhapur, India; 2 Dept of Higher
Plants, Biological Faculty, Moscow State University,
Moscow, Russia; 3 Jodrell Laboratory, Royal Botanic
Gardens, Kew, UK
Recent publications on the morphology, systematics and phylogenetic affinities of Hydatellaceae have generated deep interest in the family. The family is presently considered as monogeneric with one species in New
Zealand, ten in Australia and one in India. Trithuria konkanensis Yadav et Janarthanam was discovered and described from India in 1994. The species is common during the monsoon season on lateritic plateaus near the west coast of the country. It is widely distributed in coastal plains of Konkan, Goa and Kanara from Ratnagiri in north to Udupi in south. It grows in association with monsoon herbaceous vegetation of accumulated sandy soil on plateaus of low altitude of Malabar coast and sometimes plateaus of higher altitude in Sahyadri ranges.
It is a small moss-like plant which grows in association with Dimeria sp., Eriocaulon eurypeplon, E. maharashtrense, Ischaemun sp., Rotala densiflora ,
Utricularia caerulea , U. minutissima , U. reticulata , Xyris indica on accumulated sandy soil on lateritic plateaus from sea level to 600 m altitude. Seeds germinate sometime in June and plants show vegetative growth during heavy rain fall period from July to August end.
The species shows peak flowering during the first two weeks of September. By early October the plants and plateaus dry and seeds remain embedded in the soil till the subsequent monsoon. The only mechanism for seed dispersal is by means of soil and seeds carried by water current. The plant shows rosette habit. It produces many inflorescences at ground level. The stigmatic hairs form a rosette in the centre of plant. The stamen from each inflorescence stands erect in the centre of leaf rosette.
Anthers dehisce laterally releasing pollen grains which get deposited on stigmatic hairs below by gravitational force. Monosulcate pollen grains germinate on any cell of uniseriate stigmatic hair which acts both as stigma and style. The cells of stigmatic hairs contain red pigments and show active cytoplasmic streaming. The stigmatic hairs 2–5 in number are produced in succession and are seen at different stages of growth. Each inflorescence possesses many stalked ovaries borne below single
245

stamen in each inflorescence. The morphological nature of different structures in an inflorescence is still an enigma for botanist. The seeds remain attached to newly formed plants. Detailed observations for the last fifteen years on distribution, vegetative morphology, water stomata, phenology, pollination, cytology, cytoplasmic streaming in the cells of stigmatic hair, seed and seed dispersal in Trithuria konkanensis , the only species from
India are illustrated and presented in the present communication. In addition to this, habitat ecology and developmental morphology of T. konkanensis are compared to those of its closest relative, T. lanterna from tropical Northern Australia.
Variation in complete plastid genomes at lower taxonomic levels: a case study in
Trithuria
(Hydatellaceae, Nymphaeales)
Logacheva, M 1 , Rudall, P 2 , Graham, S 3
1 Moscow State University, Belozersky Research Institute of Physico-Chemical Biology, Russia; 2 Jodrell
3
Laboratory, Royal Botanic Gardens, Kew, UK;
University of British Columbia, UBC Botanical Garden and Dept of Botany, Canada
Recent studies of complete plastid genomes have improved our understanding of early angiosperm phylogeny, although in general they support the conclusions of earlier phylogenetic investigations based on a low number of individual molecular markers. Plastid genome studies have highlighted the fact that even a strong increase in gene sampling cannot substitute for representative taxon sampling. Therefore, a primary goal of earlier studies was to establish a representative set of complete plastid genomes covering the overall higherlevel diversity of angiosperms. In contrast, much less attention has been paid to understanding variation in complete plastid genomes at lower taxonomic levels. We are filling this gap in by comparing complete plastomes of three species of the same genus, Trithuria , which belongs to the early-divergent aquatic angiosperm family
Hydatellaceae. Although the family Hydatellaceae belongs to the basal angiosperm 'ANA' grade, being closest to Cabombaceae and Nymphaeaceae, both morphological and molecular evidence suggest that all extant members of the family (12 species in total) are closely related to each other and form a single genus. The present study deals with Australian species T. austinensis , T. australis and T. submersa , that belong to one of two principal clades of Trithuria , which we refer to as the 'temperate clade.' The phylogeny of this clade was not fully resolved in a previous molecular phylogenetic study. Incompletely resolved questions include the position of the root node of the family and the monophyly of several species. Comparing complete plastid genomes is essential in the search for informative plastid markers to clarify these and other issues, including recovery of microstructural characters for phylogenetic inference, and molecular dating efforts.
Comparative data on complete plastid genomes of three species of Trithuria will be discussed in the broader context of variation in plastid genomes across a wide range of early-divergent angiosperms investigated so far.
Rates of evolutionary transformations and the phylogenetic utility of different coding and non-coding regions of the plastid genome will also be discussed. In
246 general, we aim to distinguish between two different, though closely related goals: improving our knowledge of angiosperm phylogeny at different hierarchical levels using evidence from complete plastid genomes, and inferring general features of the evolution of plastid genomes in early angiosperms.
Dated phylogeny, character evolution and bioeography of Cyperaceae: capturing the 'missing links'
1
Muasya, M 1 , Simpson, D
A 3 , Araujo, AC 2
2 , Bruhl, J 3
, Goetghebeur, P 5
, Barrett, R 4
, Gehrke, BF 1, 6
, Gibbs,
University of Cape Town, South Africa;
Gardens Kew, UK;
Armidale;
Australia;
4
5
3
2 Royal Botanic
University of New England,
Kings Park and Botanic Garden, Perth,
Ghent University, Belgium;
Gutenberg-Universität Mainz, Germany
6 Johannes
The Cypereaceae, with about 5400 species, are among the ten largest families of the angiosperms. They have a cosmopolitan distribution, occurring mostly in open habitats and are dominant in wetlands and in habitats such as inselbergs. However, the lineages (subfamilies, tribes) differ in species richness and distribution. The
Mapanioideae (c. 200 species) occur predominantly in tropical forests while the Cyperoideae (c. 5200 species) are in open habitats. Among the Cyperoideae, the northern temperate zone is dominated by the Cariceae (c.
1900 spp.), the tropics by Cypereae (c.1100 spp.) while the southern temperate zone is dominated by Schoeneae
(c.400 spp.). The lineages differ in diagnostic taxonomic characters and key functional traits. We use a dated DNA phylogeny to infer relationships, reconstruct patterns of macroevolution of taxonomic characters (including pollen type; stem, leaf sheath/blade and inflorescence morphology) and to reconstruct the biogeography of major lineages.
Dense sampling for assessment of monophyly in
Schoeneae, a morphologically diverse tribe of
Cyperaceae
Gibbs, AK 1 , Wilson, KL 2 , Barrett, RL 3
, Musili, PM 1 , Bruhl, JJ 1
, Muasya, AM 4 ,
1
Verboom, GA 4
University of New England, Armidale, Australia;
2 National Herbarium of NSW, Royal Botanic Gardens,
Sydney, Australia; 3 Botanic Gardens and Parks
Authority, Kings Park and Botanic Gardens, Perth,
Australia;
Africa
4 University of Cape Town, Rondebosch, South
Schoeneae is nearly cosmopolitan with highest species diversity in Australia, southern Africa, south-east Asia and southern America. Plants of the tribe range from small annuals to metres tall robust perennials. A trn L-F phylogeny of the tribe supplemented by very limited sampling of ETS and ITS presented at Monocots IV in
2008 provided a starting point for assessment of

monophyly of genera within Schoeneae. Here we present a major update based on many new species sampled for trn L-F, ETS and ITS regions for better resolution and support of relationships within Schoeneae. Our study shows the value of dense sampling to avoid transient taxonomic outcomes.
Radiations and paraphyly in
Cyperus
: challenging taxonomy of a giant genus
1
Reynders, M 1 , Muasya, AM 2 , Larridon, I 1
Vrijdaghs, A 3 , Goetghebeur, P 1
, Huygh, W 1 ,
Ghent University, Belgium;
South Africa; 3
2 University of Cape Town,
Leuven University, Belgium
Calibrated family level phylogenies of Cyperaceae, revealed several origins of C4 photosynthesis as a response to changing climatic conditions during the late
Eocene (estimates between 15 and 10 mja) and seems to coincide with the origin of the savanna biome in Eastern
Africa. The ecological advantages of C
4
photosynthesis lead to a major radiative burst in Cyperus, resulting in a subclade of approximately 800 species (including 9 specialised, segregate genera). Eastern Africa can be identified as the original center of diversification of
Cyperus C
4
, from there several smaller radiations occurred within other tropical regions. Unfortunately the fast mutation rates during this radiation result in poorly resolved phylogenies even when using fast mutating chloroplast and nuclear markers and AFLP. In contrast
Cyperus C
3
lineages are well resolved and form a grade at the base of the Cyperus phylogeny. Among the segregate genera, only Kyllinga is well supported as monophyletic. Lipocarpha and Ascolepis tend to be sister taxa, however, with various levels of support. In Pycreus one large clade is well supported, however, as sister group to Cyperus laevigatus a species with dorsiventrally flattened dimerous pistils ( Pycreus has laterally flattened pistils). The other sections are nested within the Cyperus
C
4
polytomy as are the other (mostly monotypic) segregates. The presence of such a major hard polytomy, containing many convergent morphologies, forms an obstruction in building a modern classification for
Cyperus . Two opposed classification strategies are possible. Following a cladistics method all segregate lineages are to be sunken into Cyperus . Due to the high morphological diversity among its lineages, Cyperus would then only be circumscribed by cladistics arguments. The other strategy is to maintain well supported segregate lineages on the generic level and circumscribe Cyperus s.s. as a paraphyletic entity, which reflects the evolutionary processes of the group. In deciding which strategy is be the most convenient for application on Cyperus , morphological characters which have been used for generic delimitations need to be reevaluated. Shifts in seed dispersal units from nutlets to complete spikelets, in glume placements from distichous back to spiral and from trimerous to dorsiventrally flattened dimerous pistils, have been proven to have occurred several different times throughout both Cyperus
C
3
and C
4
and seem less feasible for generic delimitations. Laterally compressed dimerous pistils had been considered to be more reliable characteristics.
However, broad ontogenetic and floral vascularization studies show reorganizations of the pistil occur relatively easy throughout Cyperoideae. Even the origin of specialised inflorescences as pseudospikelets seems to be convergent for several different lineages as Lipocarpha ,
Ascolepis , Alinula , Volckiella and Kyllinga among others.
Comparing DNA-based and leaf anatomy-based phylogenies in
Carex
1
Ashton, P
Simpson, D 3
1 , North, C 1 , Smith, C 1 , Whitlock, R 2 ,
Edge Hill University, Lancashire, UK;
Sheffield, UK; 3
2 University of
Royal Botanic Gardens, Kew, UK
During the last ten years DNA-based phylogenies have been developed for large numbers of plant families and genera, including speciose genera such as Carex
(Cyperaceae), the largest sedge genus. These phylogenies have been instrumental in resolving taxonomy and ancestral relationships, but have also given a new impetus to the study of the evolution of particular characters. Mapping single traits, such as flower colour, onto molecular phylogenies to identify the number of occasions a feature has evolved is a common approach.
However, the evolutionary relationships of more complex suites of inter-dependant physical traits likely to contain a useful phylogenetic signal, but also under strong selection pressure (e.g. leaf anatomy) has been little considered. This study aims to make a preliminary investigation into this area. The form of complexes of traits can be influenced by (1) the phylogenetic or historical context provided by the species in which the traits reside, (2) the adaptations that fit the species to its environment and (3) overarching biophysical or architectural constraints that limit the traits to particular regions of morphospace (termed Seilacher’s triangle;
Seilacher 1970). Hence a phylogeny based upon DNA sequences reflects only the phylogenetic/historical dimension of trait evolution. By comparison leaf anatomy is the product of all three aspects of trait evolution. The individual traits that form complexes of traits such as leaf anatomy will also be subject, in varying degrees, to phenotypic plasticity. From an original consideration of 45 leaf anatomical characters only a number of these have been shown to be consistent within a species. Hence the characters that are phenotypically plastic have been identified and eliminated from further consideration. The remaining anatomical characters where then used to develop a phylogeny which was compared to a phylogeny for the same species based on the ITS sequences determined by
Hendrichs et al. (2004a, b). Patterns of congruence between the two trees reveal the predominance of the phylogentic signal within certain anatomical features.
Areas where the trees are dissimilar point to characteristics that are under strong selection pressure with resultant rapid evolution. Thus the study raises the possibility that the evolution of complex interrelated structures can be used to better understand phylogeny and adaptation.
Mapaniids are critical to resolving floral and spikelet ontogeny in Cyperaceae
Bruhl, JJ 4 , Vrijdaghs, A 2 , Prychid, C 1 , Reynders, M 3
247

1 Jodrell Laboratory, Royal Botanic Gardens, Kew, UK;
2 Laboratory of Plant Systematics, Institute of Botany and
Microbiology, KU Leuven, Belgium; 3 Universiteit Gent,
Vakgroep Biologie, Onderzoeksgroep Zaadplanten,
Belgium; 4 Botany, School of Environmental and Rural
Science, University of New England, Armidale, Australia
A body of modern research on floral development in
Cyperaceae based on anatomy and scanning electron microscopy, has fuelled rather than solved the debate over floral, spikelet and inflorescence character homology in the family. In Schoeneae, the debate has largely been about resolving whether the spikelets are cymose or racemose structures. In the mapaniids it has been unclear as to whether the smallest reproductive units are flowers or partial inflorescences. Different interpretations have not been resolved by the traditional methods of sectioning and SEM. Here we survey and summarise arguments and evidence to date and present preliminary findings from immuno-staining towards an evolutionary–developmental approach to resolving floral ontogeny in Cyperaceae.
Towards an online monograph of Cyperaceae
Thomas, W 1
Bruhl, J 4
, Simpson, D 2 , Roalson, E 3 , Naczi, R 1 ,
1 The New York Botanical Garden, USA;
3
2 Royal Botanic
4
Gardens Kew, UK; Washington State University, USA;
University of New England, Armidale, Australia
It is increasingly clear that a systematic understanding of large groups of plants can only be accomplished through the collaboration and contribution of specialists from all parts of the world, and those studying all lineages. To that end, the worldwide community of Cyperaceae specialists is collaborating on a global online resource for
Cyperaceae. It will provide a focus for taxonomists working on sedges throughout the world and provide information about monocots through identification keys and photos, checklists, descriptions and links to other web resources. The goal is to produce an interactive, electronic, worldwide monograph of the ca. 5400 species of Cyperaceae where we can instantly publish species descriptions, keys, specimen data, and images online as they become available. Why Cyperaceae? Sedges are one of the world’s ten most diverse families of flowering plants. They are one of the most important families in the world in terms of total biomass and ecosystem services.
The family is also remarkably biologically diverse, including 1 cm tall ephemerals, fire-resistant tussocks, scandent vines over 10 m long, and submerged aquatics.
They can be pollinated by wind or insects, and exhibit multiple emergences of C photosynthesis in different
4 genera. More than 500 species have uses as food, materials, fuels, social or religious materials, poisons, medicines, and environmental restoration. Conversely, some of the world’s worst agricultural weeds are members of the Cyperaceae. While there are useful online resources available at different sites (e.g.,
Louisiana State University and the New York Botanical
Garden), the focus of the collaboration will be through the eMonocot web resource led by the Royal Botanic
Gardens, Kew, UK. This site will provide biodiversity information such as nomenclature, taxonomic descriptions, images, identification guides, geographical,
248 ecological, DNA sequence and conservation data for all
Monocotyledons. eMonocot Pages will be generated by providing software tools to enable monocot scientists anywhere in the world to participate. Each contributor will have ownership and authorship of their contribution.
This system will support the existing collaborations and working practices of taxonomists to facilitate and manage these global interactions. Our goal is to revise the Cyperaceae for the world online, to unify and modernize the existing data, and improve our knowledge of the family, and facilitate the study and identification of sedges with web-based tools. To achieve our goals, we will (1) provide a modern classification based on molecular phylogenetic hypotheses; (2) gather and organize the vast and diffuse body of descriptions of
Cyperaceae species and related data; (3) make these descriptions, images, glossaries, and interactive keys available over the internet.
Genetic signatures, functional traits and environmental gradients: an integrated approach for investigating the distribution and assemblage of rainforest trees
1
Rossetto, M 1
National Herbarium of NSW, Sydney, Australia
The integration of molecular data with environmental and functional information assists the exploration of the micro- and macro-evolutionary processes that influence the temporal distribution and assemblage of species across the landscape. In our research we identify important factors at the single species level, test their importance in multi-species investigations, and explore their broad long-term impact across whole communities.
We take advantage of new technical and theoretical developments to update our aims and hypotheses.
Our work on over 30 woody species from subtropical and tropical assemblages (ranging from New South Wales to
Cape York) has identified some strong emerging patterns. We found that the traits relating to seed dispersal impact on the distribution of genes and individuals across the landscape. However, this impact is regionally differentiated, as suggested by re-colonisation patterns of species and assemblages. This is highlighted by genetic and distributional structure across some of the common geographical barriers we identified in our research (eg. Black Mountain Corridor, Clarence River
Corridor, Hunter River Corridor). A range of mechanisms that enable the localised persistence of individuals through temporal environmental change also impact on distributional patterns of individuals and genes. These mechanisms range from clonality to individual regeneration in the absence of disturbance, and confer individual and population-level stability. Our findings confirm the biodiversity value of the Wet
Tropics, and further highlight the importance of the subtropics (in particular Nightcap–Border Ranges).
Integrative, landscape-level studies enhance our understanding of how interactions between plant

functional attributes, and evolutionary / biogeographic processes define the distribution and assemblage of rainforest trees along environmental gradients. Coming to terms with the effects of current and historical events will increase our predictive capacity to conserve and manage rainforest communities in face of future change.
Fossil ferns from the Eocene of Argentina and the deep-time links between Southern Hemisphere rainforests
Carvalho, MR 1
Johnson, K 4
, Wilf, P 1 , Gandolfo, MA 2 , Cúneo, NR 3 ,
1 Dept of Geosciences, Pennsylvania State University,
USA; 2
3
LH Bailey Hortorium, Cornell University, Ithaca,
USA; Museo Paleontológico Egidio Feruglio, Chubut,
Argentina; 4 Denver Museum of Nature and Science, USA
In recent years, paleobotanical explorations in southern
South America have recovered an increasing number of taxa currently restricted to Australasian rainforests, highlighting biotic connectivity between these areas in the geologic past and suggesting a complex biogeographic history. The Eocene deposits of Laguna del Hunco (51.9 Ma), Patagonia, Argentina, bear remains of the oldest known Eucalyptus (Myrtaceae), as well as many rainforest-associated gymnosperm taxa including
Papuacedrus (Cupressaceae), Agathis (Araucariaceae), and Dacrycarpus (Podocarpaceae) and several angiosperm taxa such as Gymnostoma (Casuarinaceae),
Proteaceae, Atherospermataceae and Cunoniaceae.
Herein, we show that the pattern of widespread
Gondwanan distribution observed for angiosperms and gymnosperms is also evident for the pteridophyte component of this flora. Three fern species have been recently recovered and identified based on the preservation of fertile fronds and sporangial remains.
One fossil species bears acrosticoid sporangia on pinnatifid frond fragments typical of Osmundaceae.
Dicksoniaceae is recognized based on remains of fronds with marginal sori covered by a bivalved indusium, while the presence of Gleicheniaceae is confirmed by fertile fronds that have naked, round sori composed of 3-5 sporangia restricted to the upper branch of bifurcating secondary veins. Total evidence approaches based on morphology and previously sequenced chloroplast ( rbc L, atp B, rps 4) and nuclear genes (18S rDNA) were used for testing the natural affinities of these fossils. Each fossil species was scored for 25–35 characters of frond and reproductive structure morphology, rendering hypotheses of relationships of the fossils to each of the three mentioned families. The results suggest intrafamilial affinities to Todea (Osmundaceae), Dicksonia
(Dicksoniaceae), and Sticherus (Gleicheniaceae). These genera include species native to Australia and have varying number of endemic species in montane and lowland rainforests. Todea is now restricted to montane, humid subtropical and tropical Australasia and South
Africa, and has been reported for the Jurassic of
Argentina. Dicksonia is present in the wet subtropics and tropics of Australasia and South America, while
Sticherus is pantropical and austral, and mostly humid montane. Although the fossil record for Dicksonia and
Sticherus is sparse; both genera are known from
Oligocene deposits of Tasmania, and the former was previously reported from the middle Eocene of
Argentina. The records from the Eocene of Patagonia show a wider distribution for these three genera in the past, and the same long-term patterns of shared lineages between Australasia and South America observed before in angiosperms and conifers. Whether the extant distribution of fern lineages shared between Australia and South America is a product of independent colonization events, or evidence of a shared, widespread ancestral biota remains unknown. However, the warmer climate that predominated the Eocene, and the closer proximity of both South America and Australia to
Antarctica support the hypothesis of a nearly continuous forest cover throughout these landmasses. The widespread-Gondwana biogeographical pattern shared by multiple lineages in this Eocene flora suggests that an important portion of the biogeography of the Southern
Hemisphere biotas is related to processes of range expansion and local/regional extinctions of taxa in the geological past.
Assembly of the Australian rainforest flora
Jordan, G 1 , Sniderman, K 2
1 University of Tasmania, Australia; 2
Australia
Monash University,
Advances in information technology are making it increasingly possible to study whole biotas. Till now, studies of the assembly of biotas have almost all depended on small subsets of the relevant biota, using either phylogenetic or fossil approaches. These analyses are potentially biased because phylogenies are often performed on groups chosen for ideosyncratic or parochial reasons, and the fossil record can be biased by failing to show taxa that are present in any given flora.
These biases can be avoided through combining fossil and phylogenetic evidence and analysing whole biotas, or major subsets of them. We used such an approach to examine the assembly of the Australian rainforest flora, which has played a central role in understanding biome assembly processes. We analysed all identifiable disjunctions between woody Australian rainforest plant taxa and relatives on other landmasses (excluding New
Guinea). The rainforest flora shows over 1000 such disjunctions, almost half of these are within species. As evidence for regional antiquity, we evaluated the proportion of these disjunctions represented in the
Australian fossil record. Using within-species disjunctions as a proxy for recent migration, we assessed the extent of recent immigration from Asia into
Australia. To avoid problems with representation of fossil types in the fossil record, we compared fossils with fossils by contrasting Late Quaternary, Pliocene and
Oligo–Miocene Australian pollen records interpreted as tropical rainforest. We infer that eastern Australian mesothermal rainforests include some pre-Neogene clades but are numerically dominated by more recent migrants, mostly from Asia/Malesia; and that lowland tropical and 'dry' rainforests have been assembled from relatively recent migration from Asia/Malesia. The composition of mesotherm rainforest changed little between the Oligo-Miocene and the Pliocene, but the
Late Quaternary record has much greater richness of rainforest angiosperms, consistent with extensive late
Cenozoic migration in the assembly of northern
Australian rainforest flora. Our data also add support to
249

the long-established concept that the southern Australian, cool temperate (microtherm) rainforests are mainly
Gondwanan relicts.
Australian tropical and sub-tropical rainforest community assembly: functional traits and phylogenetic history
1
Kooyman, RM
M 4
1 , Rossetto, M 2
California, Berkeley;
Australia
4
, Cornwell, W 3
Botanic Gardens Trust, Sydney, Australia;
Herbarium of NSW, Sydney, Australia; 3
, Westoby,
Macquarie University /National Herbarium of NSW,
2 National
University of
Macquarie University, Sydney,
The evolutionary and biogeographic history of species and functional traits influence rainforest community assembly processes. Historical differences between latitude regions in terms of climatic stability and geographic configuration (including areal extent and topographic relief) influence rates of speciation and extinction, and have long-lasting effects on the size and diversity of regional species pools. To understand the timescales of the emergence and origin of lineages in relation to the record of global and continental geologic, geographic, and environmental change required that we place community assembly into a phylogenetic framework. Working hypotheses reflected questions about biogeography and the factors that influence evolutionary and ecological processes, and how these structure and shape species distributions, the spread of functional diversity, and community assembly.
Measuring community phylogenetic structure in relation to increasing geographic scales allowed us to investigate the extent to which niche conservatism vs. lability, and dispersal limitation operate across and within latitude regions. Positioning traits that influence community assembly across geographic scales and different landscapes relative to phylogenetic measures provided an important test of the potential role of dispersal.
Independent fossil evidence and palaeo-ecological data improved interpretations of rainforest origins, and the impacts of climatic oscillations through time.
Quantifying phylogenetic community structure, and functional trait data relative to spatial scales and environmental gradients allowed us to link current-day rainforest community assembly processes across tropical and sub-tropical latitude regions to the narrative history of Australian rainforest biogeography.
Laurasian invasion? Phylogenetics and the origins of the Australian tropical rainforest flora
Crayn, DM 1 , Costion, C 2 , Harrington, MG 1
1 Australian Tropical Herbarium, James Cook University,
Australia; 2 University of Adelaide, Australia
The origins and evolution of the Australian tropical flora and its relationship with Asian floras have fascinated botanists for over 150 years. Nineteenth to mid-twentieth century notions of Australian tropical rainforests as depauperate samples of SE Asian floras and therefore
‘alien and invasive’ have given way to hypotheses that they represent remnants of an ancient Gondwanan flora.
250
Key knowledge advances that promoted this paradigm shift include the acceptance of continental drift and the immense improvement in knowledge of the flora of the
Australian Wet Tropics (AWT) including documentation of the concentration of so-called ‘primitive’ angiosperm families and discovery of the presence of many narrow endemics (especially genera). While maintaining that
AWT rainforests are a centre of high diversity and endemism, the idea of a significant post-Miocene interchange between the Indo-Malesian and Australian floras after a long period of separation has been postulated. Dated molecular phylogenies offer unprecedented power to evaluate competing hypotheses of lineage origins. We analyze in a comparative framework published and new data for some key lineages of the AWT, including the basal angiosperms, to quantify the relative contributions of ‘Gondwanan’ and
‘Laurasian’ elements to the Australian and SE Asian tropical rainforest floras, date their origins, and to generate and test general explanations about tempo and mode of evolution of plants in this biome.
Using Lauraceae as a model system to investigate population divergence, expansion and contractions
1 across the Australian rainforest landscape
Van Der Merwe, M 1
National Herbarium of NSW, Botanic Gardens Trust,
Sydney, Australia
The family Lauraceae is a good system to investigate patterns and process of evolution within the Australian rainforests. Fossil data suggest that the family Lauraceae have been present in Australian rainforests since the origin of this vegetation type on the continent, and representatives from all major taxonomical clades of the family form part of the Australian rainforest flora.
Distribution patterns of species vary between extremely rare and uncommon to common and widely distributed, and species are found in all types of rainforests but are also not restricted to the rainforests. In this paper we discuss the results from a comparative study using DNA markers to investigate levels of intra-species molecular divergence of species with different distribution patterns alongside species level phylogenetic relationships.
Distribution patterns included in this work represent species with (A) highly disjunct distribution patterns
( Endiandra globosa and Cryptocarya laevigata) ; (B) wide distributions ( Neolitsea dealbata , Endiandra discolor , Cinnamomum oliveri and Cryptocarya triplinervis ; and (C) narrow distribution ( Endiandra introrsa and Endiandra jonesii ). Coalescence methods were used to estimate relative dates of divergence between populations and sister species. The two species with highly disjunct distribution patterns are found in areas generally believed to be rainforest refugia, one of which is in north Queensland and the other in northern
NSW. These two regions are separated by over 1000 km.
In these two areas are also found many Lauraceae endemics with narrow distributions. Two such species are Endiandra introrsa and Endiandra jonesii with the latter restricted to a small area in northern Queensland and E. introrsa found only in northern NSW. The four widely distributed species included in the study have populations that occur within the northern and southern refugial areas and the area in between. These also vary in

their distribution patterns, Neolitsea dealbata being very common and found across the entire area covered by rainforests in Australia and the other three species more restricted in their habitat. We discuss the results from the intra-species molecular study in relation to the history of the Australian rainforest, population expansion and contraction, phylogenetic relationships amongst
Australian Lauraceae and the role of aridification in speciation within the Australian rainforest flora.
A bio-geo-climatic analysis of Australian macrofungi
May, TW 1 , Klazenga, N 1
1 Royal Botanic Gardens Melbourne, South Yarra,
Australia
Biogeographic analysis of macrofungi (such as agarics, puffballs and polypores) is hampered by incomplete distribution data for individual species. A significant proportion of specimens of Australian macrofungi in
Australian herbaria has now been databased.
Nevertheless, the average number of specimens per species is only around 10, and there is a highly skewed frequency distribution for the number of specimens per species, with more than 50 percent of the 1500 species of macrofungi represented by fewer than five specimens.
Observational data is available from the Fungimap scheme, but only for 100 readily recognisable target species (with an average of more than 200 records per species). Even for better known species, detecting biogeographically meaningful patterns from mapping is complicated by the mix of habitats that occur in close proximity within what are often broad distributions spanning several states within the continent. Therefore we have developed a bio-geo-climatic classification of the distribution patterns of Australian macrofungi that produces geographic distributions of groups of species with similar climate envelopes. Specimen and observational data was pooled for species occurring in five or more 2.5 minute grids. After removing duplicate records of the same species in the same grid, a data matrix of 720 species for 19 climate variables was created (the value for each climate variable being the average across the grids from which the particular species was recorded). An association matrix was derived and subjected to clustering and ordination to produce a three and ten group classification. The three group classification corresponds well to the Bassian (southeastern and south-western), Eyrean (interior) and
Torresian (northern) distribution patterns well known for other organisms. For the fungi, however, the distribution of many species crosses the boundary between the
Bassian and Torresian areas, with distributions often extending along the entire length of the east coast.
Subgroups within each main area were also distinguished, such as two groups occurring in the semiarid interior; one (exemplified by Battarrea stevenii ) occurring in areas with annual precipitation around 500 mm and the other (exemplified by Podaxis pistillaris ) occurring in areas with much lower annual precipitation, and having a more strictly inland distribution. There is very little evidence for short-range endemism among macrofungi, of the type that is common for plants, where many species are restricted to narrow geographic ranges.
Lack of data is thus the most likely explanation for restricted geographic distributions of macrofungi, as currently known. The technique can be extended to other groups of fungi and to broader geographic areas. The biogeo-climatic patterns distinguished also provide a sound basis for further biogeographic analyses including (1) assessing the composition of mycota within each region / subregion in relation to broad geographic elements and in terms of fruit-body morphology (e.g. sequestrate forms) and (2) phylogeography, such as in relation to single or multiple transitions from one area to another. Another application of the areas that have been distinguished is for rapid data cleaning across pooled species by detection of outliers resulting from miscoding of geocode or incorrect identification of specimens.
Diversification of sequestrate fungi in Australasia: evolution and biogeography
1
Lebel, T 1
National Herbarium of Victoria, Australia
Sequestrate fungi have evolved numerous times from fungi that have epigeous fruit-bodies and forcibly discharged spores. Australasia offers an ideal locale for analysis of the diversification patterns within a phylogenetic and biogeographic framework due to high the diversity of sequestrate fungi (as many as 2450 species), and knowledge of ectomycorrhizal plant associates. Comparisons are made of sequestrate-rich lineages such as the Mesophelliaceae, Boletaceae, and
Russulaceae, and sequestrate poor Hydnangiaceae,
Inocybaceae and Entolomataceae. We explore whether diversification of the sequestrate sporocarp form occurred at multiple, disjunct points in time or in waves; appears more in some lineages than others based upon timing of initial divergence; and correlation with climatic events such as glacial cooling and drying cycles.
Similarities in the floras and mycotas between Australia, the Indian subcontinent, New Zealand, New Caledonia,
South America and Africa are well recognized. However, little is known about the truffles, because of their cryptic habit, and few workers have looked beyond their own restricted geographic area. Relationships of the truffle mycota have particular interest, because their spore dispersal requires animal vectors and is unlikely to span even relatively narrow ocean barriers. As the majority of truffles and related agarics are ectomycorrhizal, plant associations, in particular Nothofagus and Eucalyptus distributions both historical and current, may also hold some clues to biogeographic patterns.
Are the Parmeliaceae (Ascomycota) originated in
Southern Hemisphere?
Crespo, A 1 , Divakar, PK 1
1 Departamento de Biología Vegetal II, Facultad De
Farmacia, Universidad Complutense de Madrid, Spain
Biogeographical studies of lichens used to be complicated because of the large distribution ranges of
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many species. Molecular systematics has revitalized lichen biogeography by improving species delimitation and the better information it provides about species range limitations Parmeliaceae is one of the largest and widely distributed group of lichenized Ascomycota. Many species and even genera were considered to have large distribution areas extending both Northern and Southern
Hemisphere. In the last decade the family has been extensively studied using molecular tools. The molecular phylogenetic studies have demonstrated that some of the genera were monophyletic, some were not, and others, previously believed to be unrelated, fell within single monophyletic groups. Currently, there are 2500 species distributed in 84 accepted genera in the family. Most of genera are from Northern Hemisphere while the higher percentage of species is distributed in Southern
Hemisphere. However, an important number of species are pantropical distributed in both hemispheres. We used a likelihood approach to study the geographic range evolution focusing specially on pantropical groups. This analysis suggests that the ancestral range of
Xanthoparmelia , Parmotrema , Flavoparmelia , and
Austroparmelina was restricted to the Southern
Hemisphere and that subsequent long-distance dispersal is responsible for the pantropical occurrence of many species especially in Parmotrema .
Do ectomycorrhizal and saprotrophic mushrooms show different biogeographic patterns?
Hosaka, K 1
1 Dept of Botany, National Museum of Nature and
Science, Japan
One of the first molecular phylogenetic studies of mushroom-forming fungi was published by Hibbett et al.
(1997). Their results revealed several unexpected relationships, which have never been proposed in any traditional taxonomic systems based on morphological characters. One such relationship is a monophyly of the
'gomphoid-phalloid' fungi, including morphologically and ecologically diverse taxa, e.g., stinkhorns, earthstars, coral fungi, etc. The following studies (Hosaka et al.
2006, James et al. 2006) repeatedly showed the same results, and this group is now known as the subclass
Phallomycetidae. No apparent synapomorphic characters have been identified for this group, but previously neglected characters, such as rhizomorph anatomy showed some potential values. It is particularly interesting to note that four closely related orders within the subclass each show distinct morphological and ecological characters. Thus, comparative biology, including biogeography, of Phallomycetidae can potentially serve as one of the model systems in mushroom-forming fungi. Although fungi are ubiquitous and play an important role in terrestrial ecosystems, fungal biogeography (also known as 'mycogeography') has not been extensively?studied within a phylogenetic framework, except for a limited number of groups. The cryptic nature of many fungi makes them difficult to sample, and thus hindering global scale biogeographical studies. In this talk, I will examine my preliminary results from multigene analyses of Hysterangiales and
Geastrales, two of the four major orders within the subclass Phallomycetidae (Agaricomycetes,
Basidiomycota), to understand the comparative
252 biogeography of closely related, but morphologically and ecologically distinct groups. For example, unlike
Hysterangiales, which is characterized by ectomycorrhizal habit and hypogeous (below-ground) fruiting bodies with spore dispersal by animal (i.e., rodents and small marsupials) mycophagy, Geastrales is characterized by saprobic habit and mostly above-ground fruiting bodies with spore dispersal by wind.
Comparative biogeography of four closely related groups within Phallomycetidae will provide exciting insight into the fungal biogeography, which is still in a developing stage as compared to plant and animal studies. No such comprehensive biogeographical studies for macrofungi are yet available.
Historical biogeography of the
Cladia aggregata complex (Cladoniaceae, Ascomycota)
Parnmen, S 1,2 , Rangsiruji, A 1 , Lumbsch, HT 3
1 Dept of Biology, Srinakharinwirot University, Bangkok,
Thailand; 2 RAMK Herbarium, Dept of Biology,
Ramkhamhaeng University, Bangkok, Thailand; 3 Botany
Dept, Field Museum, Chicago, USA
The Cladia aggregata complex exhibits remarkable morphological and chemical disparity. Nine species are currently recognized in this complex. The group has its centre of distribution in Australasia, but some some species have a wider distribution. Cladia aggregata has a wide distribution, also occurring in Southeast Asia northwards to southern Japan and Korea, as well as India,
South Africa, Central and South Americas. Within C. aggregata alone at least six chemical races have been identified. Some of these chemotypes have been accepted as species by some authors, while other authors regard them as chemotypes. This study aimed at (i) investigating patterns of the chemical variation in the Cladia aggregata complex using molecular data and (ii) assessing genetic differentiation among evolutionary lineages. Specimens from several regions, including
Southeast Asia (Thailand and Malaysia), India,
Australasia (mainland Australia, Tasmania and New
Zealand) and the Neotropics (Brazil, Chile, Cuba and
Peru) were studied. The samples encompassed seven recognized chemotypes of the Cladia aggregata complex. The phylogenetic reconstruction of a combined data set of ribosomal nuclear ITS and the protein-coding
GAPDH gene shows at least thirteen distinct lineages that are strongly correlated with their chemotypes, and some lineages also correspond to the geographical origins. Furthermore, we applied the fixation index (Fst) to evaluate gene flow among populations. Some lineages show degrees of genetic variation thast indicate that they represent distinct lineages. Moreover, it is interesting to note that lineages which share a similar chemotype, such as beta orcinol p-depsides, show a genetic differentiation among demes. The FST-values in this group was generally high (between 0.68160 to 0.95634). The lack of genetic differentiation among the South America (Chile) and Southern Australia as well as Southeast Asia
(Penang) and Southern Australia populations within the clade indicates a relatively recent gene flow which may have resulted from more recent long distance dispersal.

Evolution of endemic fungi in New Zealand
Johnston, P 1
1 Landcare Research, New Zealand
This talk asks whether the evolution of New Zealand's endemic fungi has been driven by the same factors that have influenced the evolution of New Zealand's plants and animals. Although the New Zealand landmass was formerly part of the super-continent Gondwana, it split from east Gondwana about 80 million years ago and there are few examples of truely ancient lineages still present in New Zealand from this time. The ancestors of the majority of the present day New Zealand biota arrived over the past 30–40 million years following transoceanic dispersal. Over this same period about 15 plant families and 40 plant genera have become extinct.
A sudden drop in temperature during the past 2–5 million years has resulted in the loss of previously important parts of New Zealand's plant diversity, including
Eucalyptus , Acacia , many palms, most Proteaceae, and
Nothofagus subgenus Brassospora . Partly offsetting this loss of diversity has been the recent evolutionary opportunities provided by mountain building over the past 5–10 million years and a regular series of glacial/interglacial cycles over the past 300,000 years.
These latter events have driven the evolution of many locally endemic plant species in taxonomically rich but genetically uniform taxa such as Dracophyllum , Hebe , and the alpine Asteraceae. How has this changing plant diversity impacted upon the plant-associated fungi of
New Zealand? What have been the drivers for the evolution of those fungi which are endemic fungi to New
Zealand?
Paraphyly is a natural stage in the evolution of taxa
Hörandl, E 1
1 University of Vienna, Austria
Despite the broad acceptance of phylogenetic principles in biological classification, a fundamental question still exists on how to classify paraphyletic groups. Much of the controversy appears due to inaccurate definitions and neglection of evolutionary processes for understanding origins of natural taxa. A review of processes involved with the production of phylogenetic patterns (budding, merging, and splitting) demonstrates that paraphyly is based on shared ancestry and is a natural transitional stage in the evolution of taxa. Below species level, polyphyly transits to paraphyly and reciprocal monophyly (in the sense of holophyly); above species level, crossing barriers block polyphyly, and the evolutionary process alternates mostly between paraphyly and monophyly (in the sense of holophyly).
When a new group with shared ancestry arises it usually coexists for some time with its paraphyletic stem group.
Paraphyly and holophyly, therefore, represent relational and temporal evolutionary stages; they are two sides of the same coin. Paraphyly and holophyly, therefore, can be summarized under the term monophyly senus lato.
Paraphyletic groups exist at all levels of diversification in all kingdoms of eukaryotes, and they have traditionally been recognized because of their descent-based similarity. Since standard tree building-methods fail to reconstruct evolutionary processes other than splitting, different methodological approaches for recognition of paraphyletic groups are needed. Careful analysis is essential for discriminating paraphyly from polyphyly. In an evolutionary classification, paraphyletic taxa are potentially acceptable for classification while polyphyletic groups are unacceptable.
1
Taxonomy versus cladonomy in the dicot families
Brummitt, D 1
Royal Botanic Gardens, Kew, UK
Evolutionary taxonomy divides a phylogenetic tree into taxa based on characters related to lines of descent. These taxa are commonly given ranks (family, genus, species etc) in a hierarchical structure. One cannot divide the tree without taxa of one rank being descended from other taxa of the same rank. Recognition of paraphyletic taxa is thus essential, since the whole tree would otherwise sink into the original taxon (family, genus etc). Dividing a phylogenetic tree into only monophyletic taxa, or clades, may be referred to as cladonomy (see Taxon 1997), and is possible only if one abandons taxonomic ranks (as in the PhyloCode). The units in cladonomy are determined only by presumed lines of descent, and characters are irrelevant since they are always subordinate to lines of descent. The two approaches are theoretically incompatible, and result in quite different units of classification. Cladistic taxonomy, which has predominated for the last three or four decades, fails to distinguish between taxonomy and cladonomy, and often produces anomalous taxa because characters are ignored.
This is currently causing many upsets of well established units recognised for decades in 'traditional' evolutionary taxonomy. When characters are ignored, evolution is ignored, which is considered by many to be 'bad taxonomy'. Either one has to sink clearly recognisable taxa together or one has to split taxa where there is no appropriate character to do so. Most commonly, units in cladistic taxonomy are more broadly circumscribed than in traditional taxonomy, but in some cases families are too narrowly recognised. Examples will be given from the families of dicot flowering plants, showing circumscriptions in cladistic taxonomy which are markedly broader than in traditional taxonomy (such as
Primulaceae, Malvaceae), and other cases where excessive splitting seems to have been adopted
(Scrophulariaceae sensu lato).
1
Worldwide radiation, phylogeography and paraphyly of the speciose Rubiaceae-Rubieae
Ehrendorfer, F 1 , Vladimirov, V 2 , Barfuss, M 1
Dept for Systematic and Evolutionary Botany,
University of Vienna, Austria; 2 Institute of Botany,
Bulgarian Academy of Sciences, Sofia, Bulgaria
The tribe Rubieae with about 1000 species and a worldwide distribution includes a remarkable diversity: from
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desert shrubs, alpine and arctic perennial pioneers to tropical climbers and short-lived annuals of mediterranoid climates; from showy butterfly pollinated outbreeders to inconspicuous selfers; from relic chasmophytes to aggressive cosmopolitan weeds. Plastid intergenic DNA sequences ( atp Brbc L and rpl 32- trn L) have been studied from representatives of all major clades of Rubieae. They reveal many discrepancies between traditional phenetic classifications and allow reconstructions of all major phylogenetic differentiations and spatio-temporal radiations of the tribe. Nevertheless, relevant classifications are paraphyletic as long as most of the accepted genera are not fused into Galium sensu latissimo or further split up, both with disturbing practical and nomenclatorial consequeces. As an example: more than a dozen aberrant dioecious species from Australia and New Zealand named 'Asperula' sect.
Dioecae according to present cpDNA data are closely linked (via a still ice-free later Tertiary Antarctis?) to the near-cosmopolitan hygrophilous members of 'Galium' sect. Aparinoides , and both appear derived from the
Eurasian 'Asperula' sect. Glabella . Comparison with parallel studies by Soza & 0lmstead (2010) on various other Rubieae taxa and with other cp markers add to the reliability of current findings on the phylogeny of
Rubieae. The concept of 'phylogenetic patterns in seed plant evolution' based on aspects of life forms, reproduction strategy, cytogenetic change and differences in ecology and habitat is used for a comparative approach to the various clades of Rubieae. This should result in more general conclusions relevant to evolutionary plant systematics far beyond Rubieae.
Paraphyly in relation to endemic genera of oceanic islands and their conservation
Stuessy, T 1 , Lopez, P 1
1 University of Vienna, Austria
Oceanic islands of the world contain numerous endemic genera of flowering plants. One of the benefits of the recent surge in molecular phylogenetic analyses has been to identify more precisely the evolutionary origins of these morphologically divergent taxa. In many cases the origins lie within species-rich continental genera, hence demonstrating a type of progenitor-derivative evolutionary relationship. Classification of highly divergent island taxa (or lineages) has sometimes been at the generic level due to marked evolutionarily derived characters in island taxa that do not occur in continental progenitors. To accept these island derivatives as distinct genera, however, requires accepting the progenitor groups as paraphyletic. To some workers, this approach has been viewed as inappropriate. They favor formal taxonomic inclusion of the island endemics within the respective parental genera, thus restoring their holophyletic (monophyletic s.s.). status. The island taxa, therefore, change from being endemic genera to only being endemic subgenera, or in practice, as endemic species. This lowers the level of priority for conservation of these taxa as well as for the entire island or archipelago in which they occur. We survey the situation worldwide and emphasize the negative conservation consequences for island archipelagos through rejection of paraphyletic continental relatives and submergence of island endemic genera.
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Investigating the extent and evolutionary significance
1 of hybridisation in the New Zealand flora
Lockhart, P 1
Massey University, New Zealand
A general conclusion from the morphological studies of
Cockayne and Allan (1934) was that hybridisation is a characteristic feature of the New Zealand flora. This thinking has been used to explain paraphyletic relationships of species inferred in phylogenetic reconstructions. With the New Zealand alpine
Ranunculus, for example, paraphyly occurs in phylogenetic reconstructions for both nuclear and chloroplast encoded markers. However, while hybridisation is one possible explanation for these observations, lineage sorting also needs to be considered.
In this presentation, we describe findings using the analytical method of Joly et al. (2009) which can distinguish between these processes. We discuss the potential and limitation of this approach and also discuss predictive physiology studies using mRNA-seq analyses being used to help evaluate the evolutionary significance of hybridisation.
Polygonum
L. s.l. of Indian Himalayas: molecular phylogeny and diversification
Choudhary, R 1 , Lee, J 1
1 IBMRC, Korea Research Institute of Bioscience &
Biotechnology, South Korea
Polygonum L. s.l., commonly known as Knotweeds consist of herbs, shrubs or twinners distributed widely throughout the world but represented well in the north temperate zone. The Himalayan region is considered a second center of diversity where they occur in a wide array of ecosystems and many of which show an inclination towards speciation. To date, there has been a conflict in the classification of these Polygonums which is based on macro-morphological characters. A phylogenetic study was undertaken to assess the classification and to examine the bio-geographical history of the taxa occurring mostly in the Indian
Himalayan region like Polygonum filicaule, P. microcephalum, P. recumbens , etc. Analysis of ITS region of the nuclear DNA for 52 species provided the evolutionary relationship very well within the genus. It shows deviation from the traditional classification and supports a fresh sectional treatment with rearrangement of some species to different positions. Evident from the molecular data gathered, we assume that the diversification of this genus took place in the Himalayan region. Further, we provide a higher resolution of phylogeny to the Indian Polygonums, many of which are exclusively restricted to the Himalayas.
Bridging the gap between angiosperms and
1 gymnosperms
Wang, X 1
Nanjing Institute of Geology and Palaeontology, China

How the angiosperms came into being is a long-lasting question for biologists. Early angiosperms from the Early
Cretaceous are too young and provide no hints on the homology of carpel. Although APG have resolved most of the relationships among angiosperms based on molecular data, they shed little light on the origin of carpel and flowers. Despite works on various perspectives of flowers, the data are from the same temporal plane and the conclusions based on them are hinged to the prior assumptions. The birth of a hypothesis for the systematics of seed plants, origin of angiosperms and their carpels is triggered by the discovery of a Jurassic angiosperm Xingxueanthus sinensis . The female flower of Xingxueanthus has a free central placentation. This clashes with the current doctrines on angiosperm evolution. Comparing it with floral morphology, anatomy, and development in
Caryophyllales suggests that the placenta in angiosperms is derived from a shoot bearing ovules, and carpel from a foliar accessory, and therefore free central placentation is rather expected for early angiosperms. This is in agreement with the developmental genetic studies, molecular systematics, and fossil evidence. Cordaitaleslike plants, Coniferales, Gnetales and Angiospermae share similar reproductive structure organization. This hypothesis is well supported by cladistics using morphological data, assuming Caryophyllales as the representative of angiosperms. Flowers and angiosperms may be correlated to their counterparts in Palaeozoic gymnosperms. The debate about nature of carpel between schools is resolved by separating placenta and carpel.
With the support from concurring evidence from various fields, this new hypothesis resolves many hard problems in plant systematics, including the relationships among major seed plant groups, and demands a reorientation in character polarity in angiosperm systematic analysis. The good news from this study is that, with new assumptions, the systematics of seed plants becomes more stable and rational.
The diversification of filmy ferns in Southeast Asia
Ebihara, A 1 , Nitta, J 2
1 Dept of Botany, National Museum of Nature and
Science, Japan;
USA
2 University of California at Berkeley,
Among basal (or non-polypod) leptosporangiate fern families, Hymenophyllaceae possess several unique characteristics other than their ‘filmy’ lamina: e.g. noncordate, long-lived gametophytes which lead to the
‘independent gametophyte’ phenomenon, apogamous reproduction (in some Crepidomanes species) and drastic changes in chromosome base number (in
Hymenophyllum ). Major infrafamiliar phylogenies having been clarified, we now focus on species or lowerlevel relationships, especially on species complexes composed of morphologically ill-defined species including Vandenboschia radicans, Crepidomanes minutum and Hymenophyllum polyanthos . Our analyses combining chloroplast/nuclear DNA markers and ploidy levels successfully clarified reticulate evolution in the
Vandenboschia radicans complex in and around the
Japanese archipelago, and independent gametophytes seem to be involved in the formation of some hybrids.
The Crepidomanes minutum complex is a typical Asia–
Pacific element, and only one polymorphic species that has been recognized is distributed throughout the area.
Our analysis showed much genetic variation within the complex and several clades were recognized as in the V. radicans complex, nevertheless the morphological variation has not been fully explained by the gapCp genotypes. Southeast Asia is one of the most species-rich areas for the family, and more than one hundred species recorded there contain many potential materials for detailed studies. Most of the trichomanoid species are widely distributed throughout Southeast Asia to the
Pacific area, on the other hand, the distribution of hymenophylloid species is generally more restricted, notably that of Hymenophyllum subsp. Hymenophyllum is a group characterized by its high species richness in
Southeast Asia constituting about one third of the
Malesian Hymenophyllaceae. It has probably diversified in relation to the epiphytic habitat in montane foggyforests as is the case in the subgenus Sphaerocionium that has highly diversified in the neotropics. Though a large proportion of the filmy fern species belong to subgenus Hymenophyllum , they still remain unsampled and/or little-known in its largest source New Guinea.
Thus New Guinea material is highly essential for understanding the diversification of filmy ferns in
Southeast Asia.
The rise of the Himalayas enforced the diversification of SE Asian ferns by altering the monsoon regimes
Wang, L 1, 2 , Zhang, X-C 1 , Schneider, H 2
1 Institute of Botany, Chinese Academy of Sciences,
Beijing, China; 2 Dept of Botany, Natural History
Museum London, UK
Himalaya–Hengduan Mountains have long been noted for their exceptional species richness and high endemism.
Dating the Tree of Life has now become central to relating patterns of biodiversity to key process in Earth history such as plate tectonics and climate change.
However, a limited number of studies using phylogenetic methods have been constructed to explore the role of uplift of Himalayas and the subsequent climatic change in the diversification of plants, let alone ferns. Here, we used species-level phylogeny incorporating divergence time estimates to investigate the impact of climatic fluctuation and orographic events on diversification of
Lepisorus , a derived paleotropical fern genus. We show that the genus likely originated in eastern Himalaya plus the Hengduan Mountains in 11.67–8.66 Myr ago. We detected a signature of massive extinction, followed by a rapid radiation at about 3–2 Myr ago, when most extant clades diversified. We also detected twice increases of diversification rates in the LASER analyses. One corresponds to the initial diversification of the genus, whereas the other accounts for the diversification of the derived clades within the genus. They respectively coincide with a period of onset and intensification of East
Asian summer and winter monsoon in Late Miocene-
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Pliocene, suggesting a link between climate and biodiversity.
Taiwan as a hub of fern diversity
Chiou, W-L 1 , Liu, Y-C 2 , Chen, C-W 3
Wang, C-N 2
, Kuo, L-Y 2 ,
1 Taiwan Forestry Research Institute, Taiwan;
Chiayi University, Taiwan; 3
2 National
Taiwan Forestry Research
Institute
Taiwan, comprising the main island and its associated islets, is located near the boundary between the Holarctic and Paleotropical regions. Two-thirds of the area in
Taiwan is occupied by hills and mountains, and the elevation ranges from seaside to about 3,950 m. This complicated topography forms various habitats with different climate patterns, harboring > 4,000 vascular plants, including about 700 species of lycophytes and ferns. The lycophytes and ferns in Taiwan are diverse, with about 25% of south to Indo-China element, 17% of
Asiatic element, 16% of Himalayan element, 11% of
Malesian element, 9% of pantropical (ad hoc paletropical) element, 8% of Japan/northeastern China element, 3% of temperate element, and 11 % of endemism. The repetitive glacial and interglacial periods caused the plant migration between the Asian continent and Taiwan. The vicariance thereafter might have caused plant speciation and evolution, and together with the functions of noutheastern monsoon, southwestern monsoon, typhoon, migratory birds, and probably the jetstream and trade wind, promoted dispersal of lycophytes and ferns among the Asian continent, Taiwan and nearby islets, and paleotropical regions by their tiny spores. Athyrium (mainly distributed in temperate regions), Humata repens complex (mainly distribured in paleotropical regions), and Deparia lancea complex
(mainly distributed in eastern Asia and southern China) are given as examples illustrating Taiwan as a hub of fern diversity.
Exploring niche evolution during the colonization of
Malesia by ferns
Bystriakova, N 1 , Schneider, H 1
1 Natural History Museum, London, UK
The Malay archipelago is known not only for its species richness, especially in vascular plants, but also for its relatively young geological age. The archipelago was formed through the collision of the Australian craton with South Asia during the last ca. 10 million years. The collision resulted in the formation of islands that today harbor large numbers of plant species. The alps of New
Guinea can be the most outstanding example of a species rich area given its relatively young age (<5 mya) and high number of endemic species. The diversity found in this area today is the result of a combination of processes including colonization from different source areas, such as SE Asia and Australia, and local diversification of the colonizing lineages. The offspring of some of these colonizers can be found in the same environments as their ancestor’s whereas the offspring of others occupy niches that are filled by other taxa in the area of the origin of their ancestors. Thus Malesia, and especially the
256 mountains of New Guinea, are extremely well suited to explore the role of niche conservatism in the evolution of land plants. We will address this question using two key studies. In both studies, we use a set of analyses including phylogeneitcs, ancestral area reconstruction, divergence time estimates, climate niche modeling, and ancestral niche reconstruction. The first example, drynarioid ferns show some evidence for exploration of new niches after their arrival in New Guinea, whereas the second example, scaly tree ferns apparently display evidence for niche conservatism. We will discuss these results in the context of limitations of the methods, restrictions of our knowledge of the biology of some of these taxa, and the need for more comprehensive analyses that are taking into account a much larger fraction of the fern/ plant diversity of this region. The results of such analyses will provide us with the evidence needed to explore some of the key questions concerning the rapid assembly of species diversity in geologically young tropical regions such as New Guinea, but especially the question of whether the observed species richness is linked to diversification of new niches by colonizers as suggested by adaptive radiations. The two explored examples provide very distinct answers to support this hypothesis.
The importance of SE Asia for the Hawaiian Islands
Ranker, TA
Neale, JM 4
1 , Geiger, JMO
, Neale, R 5
2 , Sundue, MA 3 , Ramp
1 University of Hawaii at Manoa, USA;
USA; 3 University of Vermont, USA;
Gardens, USA;
Research, USA
5
2
4
Carroll College,
Denver Botanic
National Center for Atmospheric
Levels of botanical endemism in the Hawaiian Islands are some of the highest of any geographic region in the world (90% for flowering plants, 77% for ferns and lycophytes). The current high islands are located approximately 4,000 km from the nearest continent. All species migrating to the Hawaiian Islands have arrived via long-distance dispersal either through the air or in the water. Source regions for original colonizing ancestors of
Hawaiian fern and lycophyte species have been hypothesized as Indo-Pacific (including SE Asian; 48%),
Pan-Tropical (20.8%), American (11.9%), obscure
(11.1%), Boreal (4.4%), and Austral (3.7%). Over the last decade multiple molecular phylogenetic studies have been conducted that have included both Hawaiian and non-Hawaiian species. The combined results of these studies provide strong support for about 50% of
Hawaiian fern lineages being of Indo–Pacific/SE Asian origin. Recent studies have also speculated on specific transport pathways via which fern spores may have been transported to the Hawaiian Islands to affect colonization. Potential proposed pathways include the sub-tropical jet stream, shifts in the Hadley Circulation–
Intertropical convergence zone, easterly trade winds, and individual storms from Mesoamerica. The first three possibilities represent a simplified view of atmospheric flow, but are unlikely to represent the actual trajectories of individual spores. The transport of fern spores in the atmosphere can be viewed as analogous to the transport of inert tracers (e.g., dust particles) studied using meteorological models, since the particle sizes are approximately the same. We will show initial efforts to

utilize an existing trajectory and dispersal model – used extensively in pollution studies – to test the probability of spores originating from the potential source regions reaching the Hawaiian Islands. Our results show the likely importance of the sub-tropical jet stream for transporting spores from the Indo-Pacific/SE Asian region to the Hawaiian Islands, which could account for
50% of Hawaiian fern lineages originating from that region.
Biogeography of the Australasian and South-west
Pacific fern floras
Perrie, L 1 , Ohlsen, D 2 , Bayly, M 2 , Brownsey, P 1
1 Museum of New Zealand, New Zealand; 2 The University of Melbourne, Australia
The fern floras of Australia, New Zealand, and some of the Pacific Islands are relatively well-characterised at the species level. New Zealand, for example, has c. 200 indigenous fern and lycophytes species. Despite half the species being endemic, there is little endemism at the generic level. Some 90% of New Zealand’s non-endemic species are shared with south-eastern, temperate
Australia. This is principally attributable to long-distance dispersal. While New Zealand lacks strong geographic centres of fern diversity, Australia’s c. 440 indigenous ferns and lycophytes (c. 40% endemic) are concentrated along the eastern coast, which can be in turn be partitioned into southern temperate and northern tropical elements. The latter has significant affinity north to
Malesia and east to the Pacific. Despite their smaller landmasses, some of the Pacific Islands (e.g., Fiji) have more fern species than New Zealand, but their species endemism is much lower, with many Pacific fern species having wide-ranging distributions. Complementing measures of shared and endemic species, phylogenies provide another dimension for assessing relationships amongst regional floras. Detailed DNA-based phylogenies are now available for several fern families.
These allow assessment of how many of the 'principal' clades of a particular family occur in each of Australia,
New Zealand, and the Pacific Islands, alongside determination of the geographic distribution/affinity of those clades. For instance, the clade dominant in the
Aspleniaceae flora of tropical Australia is different to that prevalent in south-eastern Australia and New
Zealand. These analyses of the infra- and extrarelationships of the Australasia/Pacific fern floras allow the extent of 'endemic' radiation to be determined, in terms of both species numbers and geographic scale.
Australasia and the south-west Pacific appear to be both a source and 'sink' for global fern biodiversity.
Ancient genome duplications and glucosinolate evolution in the Brassicales
Schranz, E 1
Dam, N 3
, Hofberger, J 1 , Pires, C 2 , Edgar, P 2 , van
1 University of Amsterdam, The Netherlands; of Missouri, USA;
Netherlands
3
2 University
Radboud University, Nijmegen, The
Ancient whole genome duplication (WGD), or paleopolyploidy, and subsequent gene and pathway divergence have likely played profound roles in generating the complexity of life on earth. However, we have limited examples of how particular ecological relevant traits have evolved as a consequence of paleopolyploidy. We illustrate the link between WGDs and key-trait evolution by investigating the glucosinolate defense pathway in the plant order Brassicales. The analysis of the Arabidopsis genome revealed evidence of three WGDs in the evolution of the Brassicaceae. The most recent event is called the At-
α
(alpha) and is unique to the Brassicaceae. The intermediate event is referred to as the At-ß (beta), which is shared with at least the
Cleomaceae but is absent from Papaya within the
Brassicales. The oldest is the At-
γ
(gamma) and is shared with all Rosid and potentially Asterid species. The major chemical defenses in the Brassicales are the sulfurcontaining glucosinolates. Large differences in glucosinolate profiles exist with much of the variation due to four classes of amino acid precursors.
Glucosinolates derived from Val, Ile, Leu, or Ala are aliphatic, those from Phe or Tyr are aromatic, and those from Trp are indole glucosinolates. The fourth class, the
Met-derived glucosinolates, is unique sub-set of aliphatic glucosinolates to the most speciose family, the
Brassicaceae. We provide chemical and molecular data supporting the hypothesis that the creation of the divergence of the glucosinolates is coupled to WGDs.
Using the wealth of new and emerging Brassicales genomics data we are identifying homologs of key regulatory and enzymatic genes from the glucosinolate pathway. Ongoing molecular evolutionary and gene expression analysis support that retained duplicate copies of key genes has contributed to glucosinolate diversification, specifically that the appearance of the indole pathway coincides with the At-
β
event and that
Met-derived compounds coincide with the At-
α
event of the Brassicaceae. Glucosinolates are known not only to have affects on insect herbivores, but also fungi, other plants and human health. Hence, our research should benefit ongoing and future efforts to alter glucosinolate profiles in Brassicales crop species.
Brassicales Map Alignment Project (BMAP): toward
1 a comparative omics platform
Pires, JC 1
Division of Biological Sciences, Christopher S Bond
Life Sciences Center, University of Missouri, USA
An exciting and emerging paradigm in the field of genomics is the comparison of genomes from multiple species within a genus or family (12 Drosophila genomes, Oryza Map Alignment Project). As sequencing costs plummet, it will be possible to establish comparative research platforms based on high quality reference genomes for virtually all crop plants. This will permit the acquisition of the majority of allelic diversity and allow for the systematic analysis of molecular evolution within a genus/family. Over the last year, four workshops were held to establish an international
257

collaborative network to formulate a roadmap for the generation of a comparative genomics platform for the
Brassicaceae. A summary will be presented of international meetings in China, Japan, Germany, and the
USA that discussed a set of grand challenge questions and a prioritized list of species to be sequenced. Specific arguments to justify a focus on the Brassicaceae include: the mustard family contains many important food and biofuel crops, and weed species; the plethora of novel traits and chemistries important to human health and crop improvement; the strong and vibrant Arabidopsis thaliana research community as major stakeholders; the relatively small genomes in the family amenable to current sequencing methods; the rich knowledge of karyotype evolution and chromosome dynamics in the family; the ability to bridge comparative genomics to systems biology, as a pioneer family to develop comparative systems biology, making use of the rich knowledge from Arabidopsis ; and the interest in extending systems biology of plants to rhizosphere/metagenomics
VEGI: Value-directed Evolutionary Genomics
Initiative
Bureau, T 1 , Blanchette, M 1
Stinchcombe, J
Schoen, D 1
2 , Dewar, K 1
, Wright, S 2 , Naseem, A
, Harrison, P 1 , Moses, A
1
2
,
,
1 McGill University, Montreal, Canada;
Toronto, Toronto, Canada
2 University of
In general, genomics has been traditionally centered around the notion of identifying 'host' genes that can be gleaned from genomic sequence and the subsequent high throughput characterization at the level of their gene expression and/or, if any, protein products. But host genes make up only a small percentage of many eukaryotic, including plant, genomes. Instead host genes are surrounded by a vast ocean of so-called non-coding
DNA. Long thought to be of little functional significance and not amenable to characterization by traditional genomics approaches, recent evidence has surfaced indicating that the non-coding DNA harbours islands of sequences that may have profound functional significance. We argue that this large and uncharted region of plant genomes is rich with potential sequences of developmental significance and, importantly, agronomic application. Furthermore, our proposed research project will allow the rapid harvest of the 'low hanging fruit' entering a pipeline for experimental and utility validation. We intend to reveal and characterize these functional non-coding DNA sequences by employing a strategy that combines comparative, population and functional genomics approaches. This strategy involves not only computational or bioinformatics approaches but also genome sequencing, expression profiling, gene disruption via insertion mutagenesis and RNA interference, and detailed phenotype characterization. Key to our strategy is using comparative and population genomics methodologies that uncover non-coding DNA sequences that evolve under negative (purifying) or positive (adaptive) selection. Whereas negative selection indicates sequences that are functionally conserved, positive selection indicates sequences that have novel advantageous function. The other key component of our strategy is
258 extensive experimental characterization of triaged targets. In consultation with our agricultural economics team members and scientific advisory board candidate targets will be selected that have the greatest potential for agronomic impact and, therefore, be candidates to enter our processing pipeline. VEGI focuses on the species within the family Brassicaceae, which includes such crops as canola, cabbage, and radish. Venturing into any market in the Northern Hemisphere, including those in
Canada, U.S., Asia and Europe, one quickly appreciates the importance of Brassicaceous crops. Also within the
Brassicaceae is 'the' model plant, Arabidopsis thaliana since no other plant has as many associated fully developed genomics resources. Our strategy not only has application to many members of the Brassicaceae, but fully exploits these resources in Arabidopsis . VEGI speaks directly to the Genome Canada strategic mandate in plant genomics and of improving crop plants by providing value added to plant genomics resources and facilitating the exploitation of novel regulatory regions and genes underlying traits of agronomic importance to the Canadian economy. Although our research focus is not directly on crop plants, our approach is to use more tractable systems to identify conserved functional elements that should translate into crop systems.
Evolution of sex chromosomes in Papaya and relatives
Ming, Ray 1
1 University of Illinois at Urbana-Champaign, USA
Papaya is a rare trioecious plant species with male, female, and hermaphrodite sex forms. Sex determination in Papaya is controlled by a pair of nascent sex chromosomes that contain a male-specific region (MSY) with recombination suppression. There are two slightly different Y chromosomes, Y for males and Yh for hermaphrodites, and all combinations of the Y and/or Yh chromosomes are lethal. Female Papaya has the XX genotype. We investigated the features of paired dioecious X- and Y-specific sequences and compared them to corresponding gynodioecious X- and Yh-specific regions. Numerous chromosomal rearrangements were detected between the X- and Y-specific BACs, including inversions, deletions, insertions, and duplications. DNA sequence expansion was documented on the Y and Yh chromosomes. Dioecious and gynodioecious X-specific
BACs were virtually identical. The Y- and Yh-specific
BACs shared high degree of DNA sequence identity, but local chromosomal rearrangements were detected, as the consequence of suppression of recombination in the male specific region and the isolation of Y and Yh chromosomes enforced by the lethal effect. Analysis of sequence divergence between X/Y and X/Yh gene pairs resulted in the estimated ages of divergence from 0.6 to
2.8 million years, supporting the hypothesis of a recent origin of the papaya sex chromosomes. The estimated age of divergence between Y and Yh chromosomes was
73,000 years, indicating that Y and Yh chromosomes evolved from a common ancestral Y chromosome, prior to the origin of agriculture. Papaya X/Y gene pairs were used as tools to invesitigate whether sex chromosomes have evolved in Vasconcellea , a sister genus of Carica .
Six X/Y gene pairs were cloned, sequenced, and analyzed from three dioecious, one trioecious, and one

monoecious species of Vasconcellea . The isolation of distinctive X and Y alleles in dioecious and trioecious species of Vasconcellea demonstrated that sex chromosomes have evolved in this genus. Distinctive clusters of X/Y alleles were documented in V. parviflora and V. pulchra for all available gene sequences and in V. goudatinana and V. cardinamarcensis for some X/Y alleles. Carica and Vasconcellea sex chromosomes might have originated from the same autosomes bearing the X allelic form that still exists in the monoecious species V. monoica , and evolved independently after the speciation event that separated Carica from Vasconcellea . Within
Vasconcellea , sex chromosomes evolved at the species level at least for some species.
Genome alterations and whole-genome duplications in the evolution of crucifers
Lysak, MA 1,2
1 Masaryk University, Czech Republic; 2 CEITEC MU
Since 2000, extensive genomic resources made crucifers an ideal system for comparative studies on plant genome evolution using phylogenetic, genetic and cytogenomic tools. Comparative (cyto)genetic mapping disclosed astonishingly high levels of chromosome and genome collinearity across the core Brassicaceae. These data allowed to infer ancestral karyotypes and to reconstruct the evolution towards the extant genomes as lineage- and species-specific alterations of ancestral genomic blocks.
Ancestral Crucifer Karyotype (ACK) with eight linkage groups (n=8) was established as an ancestral genome of the lineage I and most likely also of lineage II through derived PCK karyotype with seven chromosomes (n=7).
In some crucifer groups and species, ancestral karyotypes remained remarkably stable through the time, whereas other taxa manifest karyotypic variation due to chromosome rearrangements and polyploidy (wholegenome duplications, WGD). The significance of genomic and cytogenetic signatures (e.g., a unique karyotype structure) for resolving some phylogenetic controversies in the family will be demonstrated. It is becoming increasingly clear that genome complexity, cladogenesis, species richness and karyotypic variation are promoted and affected by multiple waves of differently aged WGD events followed by genome fractionation. The history and evolutionary impact of lineage-specific WGDs in some crucifer groups (e.g.,
Biscutelleae, Brassiceae, Camelineae) will be discussed.
Cryptic speciation and speciation genes in Arctic diploid plants, with emphasis on
Draba
(Brassicaceae)
1
Gustafsson, ALS
Gusarova, G 1
1 , Parisod, C 2
, Borgen, L 1
, Schranz, ME
, Brochmann, C 1
2
3 ,
National Centre for Biosystematics, Natural History
Museum, University of Oslo, Norway; Evolutionary
Botany, University of Neuchatel, Switzerland; 3 Institute for Biodiversity and Ecosystem Dynamics, University of
Amsterdam, The Netherlands
The Arctic flora has long been considered depauperate as compared to other regions of the world. However, estimates of species diversity are based on morphological differentiation (taxonomic species) and may not provide accurate numbers of biological species. In recent experiments, crosses within each of three diploid, circumpolar species of Draba (Brassicaceae) resulted in mostly sterile hybrids, suggesting that formation of cryptic biological species may be common in Arctic plants (Grundt et al. 2006). The aim of the present study is to provide further insights into patterns and processes underlying speciation in the Arctic . We currently perform intraspecific crosses within several diploid species sampled in Alaska, Canada and arctic Norway. The fertility of hybrids will enable us to draw conclusions about the generality of sibling species formation in the
Arctic. We compare selfers vs. outcrossers to address the underlying evolutionary forces (drift vs. selection), expecting selfing to facilitate the accumulation of hybrid incompatibilities through genetic drift . We also aim at unravelling more precise molecular mechanisms underlying reproductive barriers in Draba nivalis , as a follow-up of Skrede et al. (2008). QTL analyses are made on F2 hybrids from reciprocal crosses by associating seed set and pollen fertility with a range of molecular markers (‘Brassicaceae Building Block’ markers, microsatellites, AFLPs and SSAPs). We ask to what extent chromosomal rearrangements and nuclearnuclear and cytonuclear incompatibilities contribute to intrinsic reproductive barriers and aim to characterize putative speciation loci . Brassicaceae Building Block markers can offer great comparative strength. These 24 conserved genomic blocks can give additional QTL markers, and also enable us to reconstruct the Draba genome evolution by investigating the reshuffling of these blocks from the ancestral n=8 karyotype. The
Brassicaceae Building Block markers also show great potential as universal markers for the entire Brassicaceae family.
Tribal relationships and generic delimitation in the
1
Brassicaceae
Al-Shehbaz, I 1
Missouri Botanical Garden, St Louis, USA
Substantial advancements in the systematics and phylogeny of the mustard family Brassicaceae
(Cruciferae) have been achieved during the past six years that followed the introduction of the first phylogenetic tribal classification of the family during the 2005
International Botanical Congress in Vienna. During that six-year period, the number of monophyletically delimited tribes has almost been doubled from the initially introduced 25 tribes. The presentation will review these tribes, discuss their characterizations, and show their affiliations to the four major Brassicaceae lineages currently recognized. A review of unigeneric or oligogeneric tribes will be addressed, and examples to rectify the difficulties in the identification of genera and tribes across the entire family will be introduced. Aspects of the family origin, age, and diversification will be summarized, and problems relating the basal polytomy of the family will be discussed. The predominant occurrence of homoplasy, compared to the very rare
259

development of unique characters in the family, will be briefly surveyed, and evaluations of various morphologies in the Brassicaceae will be presented based on the latest evo-devo approaches on character evolution in the model organism Arabidopsis thaliana and other newly emerging model species in the family. A survey of the currently accepted genera and their tribal affiliations will be made in comparison with those recognized during the past Botanical Congress. Although nearly 95% of the genera of the family have been surveyed for at least one molecular marker, several problems remain to be resolved, including those genera not yet placed in tribes, the use of additional markers, the challenges of wholegenome analyses, and the utilization of morphological characters in the establishment of phylogenies separately or in combination with molecular data. Despite these difficulties, the Brassicaceae stand as one of the best examples of well-studied large plant families both taxonomically and molecularly. Finally, future prospects and challenges in the systematics and evolution of the family will be presented.
Principles in crucifer evolution: genome size, polyploidization and radiation
1
1
Koch, MA 1 , Schmickl, R
Heidelberg Institute of Plant Sciences, Germany
Crucifers (Brassicaceae, Cruciferae) are a large family comprising some 338 genera and approximately 3,700 species. The family includes important crops as well as several model species in various fields of plant research.
Meanwhile, we have nearly complete coverage for a molecular-systematic characterization of all genera. And, in addition numerous phylogenetic hypothesis based on various genes from the plastome, nuclear and mitochondrial DNA are available enabling us to provide first robust and reliable family-wide phylogenetic trees.
Further evidence is coming from an incresing number of whole-genome sequenced. The Brassicaceae are characterized by frequently occurring hybridization and polyploidization, which, as a consequence, is greatly affecting genome size and structure, but also any mode of speciation. Nonetheless, it is still unclear if the evolution of the Brassicaceae on the various taxonomic/temporal levels and of its evolutionary lineages is mostly or even best explained by multiple radiation events. A fact that can also account for the difficulties to resolve deep phylogenetic relationships within the family. Herein we are aiming to use the actually available knowledge and phylogenetic data and a robust phylogenetic backbone to address these principles questions of crucifer evolution by combing data on chromosome numbers, genome sizes, major whole-genome duplication events (WGD) data across the entire family to identify general patterns in crucifer evolution and seeking for chronological and geographical correlations of speciation and radiation processes.
Breeding systems in Brassicaceae
Owens, S 1 , Miller, R 1 , Al Shehbaz, I 2
1 Royal Botanic Gardens, Kew, UK;
Garden, St Louis, USA
2 Missouri Botanical
260
Breeding system data on more than 200 species of
Brassicaceae (Cruciferae) has been extracted from over
700 literature references. The results indicate that the variety of breeding systems among species are variable with a range from obligate outbreeders through to obligate inbreeders. 3 species in the New Zealand genus
Lepidium are dioecious. Hirschfeldia incana and the
New Zealand endemic Cheesemania wallii are gynodioecious. Apomixis is present in the taxonomically difficult genus Boechera . 44 species are recorded as selfincompatible, 80 species are self-compatible and 5 species have self-incompatible and self-compatible individuals reported for different populations within the species. Self-incompatible species are widely regarded as possessing a sporophytic self incompatibility system.
Pseudocompatibility has been recorded several times in differing species and genera. Given the frequency of the occurrence of self-incompatibility, the robustness of the data reported on self-incompatibility and selfcompatibility in the published papers has been tested using criteria presented in this publication. The result indicates that few reports can be considered sufficiently robust. The majority of references to self-incompatibility and self-compatibility focus on data from three species namely Brassica oleracea and more recently Arabidopsis lyrata and Arabidopsis thaliana . Any significance to the distribution of breeding systems among the tribes and genera is difficult to judge as the data is both sparse and the taxonomy of the family is poorly resolved. While the evolution of self-incompatibility is discussed, the lack of both data and a robust taxonomy precludes any firm conclusions.
Phylogenetic relationships and microevolutionary patterns in
Alyssum
(Brassicaceae)
Marhold, K 1,2 , Španiel, S 1 , Zozomová-Lihová, J 1
1 Institute of Botany, Slovak Academy of Sciences,
Bratislava, Slovak Republic; 2 Dept of Botany, Charles
University, Praha, Czech Republic
Alyssum is one of the largest genera of the Brassicaceae family. In its wide circumscription it consists of about
195 species worldwide and 70 species occurring in
Europe. In spite of the wide distribution of the genus, its high species diversity, large ecological amplitude, morphological and cytotype variation, Alyssum belongs to one of the least studied genera of Brassicaceae. Only scant data exist on the phylogenetic relationships within the genus, but already a small sample of species included in the studies devoted to the phylogeny at the family and tribe (Alysseae) levels suggests that Alyssum is polyphyletic. Our study based on ITS sequences indicates a monophyletic origin of Alyssum sect.
Odontarrhena , while A.
sect. Alyssum , sect. Psilonema and sect. Gamosepalum , as currently circumscribed, will require revision. Our investigations are focused on the A. montanum – A. repens group, a highly variable species complex with controversial species circumscriptions and infraspecific classifications. Morphological, genetic
(AFLPs, cpDNA sequences) and cytotype variation analyses were performed with an aim to understand microevolutionary patterns and speciation processes in the group, and to derive a sound taxonomic concept.
Three ploidy levels have been discovered; diploids and tetraploids are common and sympatric, while hexaploids

are rare and scattered.
A. repens , represented by diploids and tetraploids, is confirmed to be a distinct species.
Within A. montanum , recognition of two subspecies traditionally based on habitat preferences, A. montanum subsp. montanum (rocky sites) and subsp. gmelinii
(sandy sites) is not supported. Most Central European populations of A. montanum form a 'core' group consisting of diploid and tetraploid populations that display little genetic variation, suggesting past bottleneck and recent (postglacial) colonization of this area.
Populations distributed south of the Alps and in the
Apennine and Balkan Peninsulas are much more diverse.
Pointing out a few examples, the hexaploid A. montanum subsp. pluscanescens , a Slovenian and Croatian narrow endemic, is well differentiated and suggested to be of an allopolyploid origin. The Apennine populations
(traditionally classified as A. montanum and A. diffusum ) are found to be genetically distinct from those from
Central Europe, and harbour high and geographically structured genetic variation. The populations from different geographic regions have apparently been isolated for long times, most likely representing descendants of multiple refugial populations and being an example of on-going allopatric speciation. Their subspecific treatment within A. diffusum is proposed.
Furthermore, populations from Switzerland and SW
Germany (including the type population of A. montanum ) are morphologically and genetically well-separated from the rest of the Central European 'core' populations, indicating that the name A. montanum should be applied in a more restricted sense than before. Overall, combination of morphological, genetic and karyological data is shown to be a powerful tool for disentangling microevolutionary patterns in polyploid complexes.
Pachycladon
, evolution of an alpine radiation in New
1
Zealand
Heenan, P
Lockhart, P 2
1 , Bickford, C 1
, Lysak, M 3
, Voelckel, C 2 , Becker, M
Landcare Research, Lincoln, New Zealand;
University, Palmerston North, New Zealand; 3
2
2 ,
Massey
Masaryk
University, Brno, Czech Republic
Pachycladon is related to the model plant Arabidopsis thaliana . This has provided the impetus for a research programme elucidating the evolution and radiation of alpine species of Pachycladon in the Southern Alps of
New Zealand. Phylogenetic analyses reveal the allopolyploid origin of Pachycladon within Lineage I of the Brassicaceae, where it is derived from hybridisation between members of the tribes Camelineae and
Smelowskieae. The Pachycladon chromosome structure confirms the allopolyploid origin, as its chromosome karyotype originated through an allopolyploidization event between two genomes structurally resembling the
Ancestral Crucifer Karyotype. Transcript profiling has been utilised as a means for identifying candidate genes potentially important in species diversification and to predict physiological processes that may have evolved differently in the genus. Differential expression of genes is shown among several Pachycladon species for glucosinolate biosynthesis and hydrolysis (e.g., ESP gene cluster), the interconversion of carbon dioxide and bicarbonate (e.g., carbonic anhydrase genes), and water use efficiency. Ecophysiological studies of the shaded alpine bluff habitats occupied by Pachycladon species are notable for the low values for photosynthetically active radiation (< 100 µmol m -2 s -1 ) and atmospheric vapor pressure deficit (< 0.5 VPD). Additional data will be presented showing how the ecophysiological processes and the genetic determinants of the
Pachycladon adaptive radiation vary across the species.
A bicontinental hybrid origin of polyploid
Australian/New Zealand
Lepidium species
(Brassicaceae)
Mummenhoff, K 1
Dierschke, T 3
, Grimmelmann, K 1 , Lysak, M 2 ,
1 University of Osnabrueck, Biology Dept, Botany,
Germany; 2 Masaryk University, Dept of Functional
Genomics & Proteomics, Institute of Experimental
Biology, Masaryk University, Czech Republic; 3 Monash
University, School of Biological Sciences, Clayton
Campus, Australia
Lepidium s.s. (Brassicaceae) (c. 200 species) is distributed worldwide with endemic species on every continent, except Antarctica. Incongruence between chloroplast and nuclear DNA phylogenies, and single additive nucleotide positions in internal transcribed spacer (ITS) sequences of polyploid Australian/New
Zealand (NZ) Lepidium species indicate their bicontinental hybrid origin. This pattern was explained by two trans-oceanic dispersals of Lepidium species from
Africa and California, and subsequent hybridization followed by homogenization of the ribosomal DNA sequence either to the African (A-clade) or to the
Californian ITS-type (C-clade) in the two different ITSlineages of Australian/NZ Lepidium polyploids. The nuclear genomes of African and Australian/NZ C-clade species were detected by GISH (genomic in situ hybridization) in allopolyploid Australian/NZ Lepidium species of the A-clade, supporting their hybrid origin.
The presumed hybrid origin of Australian/NZ C-clade taxa could not be confirmed. Hence, it is assumed that ancestral Californian taxa experienced rapid radiation in
Australia/NZ into extant C-clade polyploid taxa followed by hybridization with African species. As a result, Aclade allopolyploid Lepidium species share the
Californian chloroplast type and the African ITS-type with the C-clade Australian/NZ polyploid and African diploid species, respectively. Chromosome counts, flow cytometric DNA ploidy analysis and crossing experiments between putative parental taxa confirm this view. Artificial F1 hybrid plants do show additive nucleotide positions at all diagnostic positions.
Cryptic lineage diversity in California mosses –
2
1 examples from
Syntrichia
Fisher, KM 1 , Mishler, BD 2
California State University, Los Angeles, USA;
University of California, Berkeley, USA
261

The California Floristic Province is a biodiversity hotspot, boasting approximately 5,500 native vascular plants, with 40% endemic to the region. In contrast,
California is home to about 608 native mosses, of which only 19 (3%) are endemic. Several explanations could account for this apparent discrepancy in the endemicity of California’s vascular and moss floras. First, it is possible that this is a real biological discrepancy reflecting the potential higher dispersal rate of mosses and their supposed propensity for displaying more relictual distributions compared to vascular plants.
Alternatively, this discrepancy in endemicity may instead be an artifact of our current knowledge: either additional endemics exist but await discovery, or we currently apply a species concept to bryophytes that underestimates the actual moss diversity present in California. Here we will explore these alternate explanations for bryodiversity patterns in California using examples from Syntrichia ruralis and Syntrichia caninervis , two mosses found in arid regions of the world, including California. Fine-scale phylogenetic analysis of S. ruralis from western North
America and patterns of genetic diversity at multiple spatial scales in S. caninervis from Southern California and the Mojave Desert illustrate the need to carefully consider the unique biology of mosses when applying species concepts to this group.
Pleistocene climate shifts and southern African biomes: antagonistic interactions, bryophyte gene genealogies, and a 'dynamic islands' model of population fluctuation
Hedderson, T 1
1 University of Cape Town, South Africa
The Cape Floristic Region (CFR) of South Africa comprises the most diverse temperate flora on the planet, and the mechanisms responsible for generating this richness have long been debated. In this topographically complex area, dominated by the Cape Fold Mountains, wetter mountain habitats interdigitate with drier karoo and renosterveld vegetation, whilst forests occur only in the wettest areas. Lithological variation permits additional specialisation, with nutrient-poor quartzitic sandstones, nutrient-rich shales, and limestones possessing largely non-overlapping floras. Such complexity may influence species genetic structure by determining migration patterns. In lowland taxa gene flow is impeded by mountain ranges, whilst for upland taxa valley habitats may limit gene flow. Climatic oscillations, like those associated with Pleistocene glacial–interglacial cycles, may allow altitudinallyrestricted species to escape periodically from high altitude refuges and re-establish genetic communication with other populations. The restriction of species to refugia during periods of altered climate generates two predictions for genetic pattern: (1) such species should show stronger population differentiation than species that are less restricted; (2) populations situated in long-term stable refugia should show greater allelic diversity than those in non-refugial areas. In the CFR, climatic fluctuations coupled with topographic and edaphic variation, are likely to have altered the ranges of plants and indeed of whole biomes. In effect, high altitude habitats may be seen as constituting a dynamic system of islands, with complex temporal patterns of connection
262 and isolation. Because of the edaphic link, there may be few opportunities for lowland taxa to extend their ranges altitudinally, but for the most part there appear to be corridors between valley basins. If other vegetation types occupied these, basin systems may also have formed a dynamic series of edaphic islands during periods of climate change. The complexity of the mountain-valley systems of the region therefore offers a mechanism for the origin of new species, particularly in altitudinallyrestricted taxa. However, the joint impact of topography and climatic fluctuation on population differentiation has received little study in the CFR, despite its potential importance as a driver of genetic and species diversity here We evaluate the Dynamic Islands model proposed above through phylogeographic techniques and analysis of genetic structuring in sets of bryophyte taxa representing a) high altitude fynbos/montane habitats, b) karroo/renosterveld habitats and c) afromontane forests.
These evaluate the interaction of topography and climate in shaping population genetic structure, migration routes and speciation in representative groups from these three important biomes. The analyses reveal patterns of genetic structuring that correspond strongly to landscape features, and the timing of major splits corresponds well with inferred Pleistocene climate change. The data also reveal that interactions among the three biomes are strongly antagonistic. Besides informing our understanding of diversification processes in the CFR, the insights gained from this study have direct relevance for the management of the floristic diversity of the region. Patterns of genetic diversity within species facilitate the identification of long-term, climatically stable refugia, which may become increasingly important as conservation nodes in the face of global climate change.
Molecular population genetics of the sex chromosomes in
Ceratodon purpureus
McDaniel, S 1
1 Dept of Biology, University of Florida, Gainesville, USA
It is now widely recognized that the geographic distribution of genetic variation is not uniform across the genome of a species. Different patterns of recombination, mutation, natural selection, and genetic drift cause strikingly different patterns of polymorphism at different loci in the same organism. The moss Ceratodon purpureus , because of its haploid-dominant life cycle, developing genomic resources, and ease of collection and laboratory cultivation, is an emerging model system for studying the effects of variation in population genetic processes on patterns of molecular variation. C.
purpureus is common on disturbed soils in temperate parts of the Northern and Southern Hemispheres.
Previously we used differences in patterns of polymorphism at three loci to infer that migration among geographically distant populations was ongoing. We have now used a small EST library to develop new loci to provide a more complete understanding of the roles of different population genetic processes, in particular recombination and sex-specific demography, in shaping patterns of genetic variation. We designed primers to amplify intron-containing regions of the ESTs, and genetically mapped 70 of the ESTs in a cross between two laboratory isolates (R40 and GG1). Here I will

compare inferred patterns of molecular evolution and biogeography in sex-linked loci to molecular evolutionary patterns in autosomal loci. Sex chromosomes are widely reported among dioecious bryophytes, like C.
purpureus . Sex chromosomes presumably arose from an ancestral pair of autosomes that ceased recombining over a portion of their length following the evolution of genetic sex determination. As a consequence of the bryophyte life cycle, recombination is suppressed on both the male and female sex chromosomes. Thus, both sex chromosomes are expected to show the deleterious effects of suppressed recombination, but purifying selection should act to maintain gene function in both males and females because genes on both sex chromosomes are expressed in the haploid gametophyte stage. Any differences in molecular evolutionary patterns between the sexes therefore arise from sexually dimorphic patterns of demography, mutation, or selection. I tested these hypotheses using molecular population genetic analyses of the several sex-linked loci in the moss C. purpureus .
Both male and female sex linked loci showed almost no nucleotide variation, consistent with other sex chromosome systems. Both male and female loci exhibited similar biogeographic patterns but lower population structure than autosomal loci, suggesting that selective sweeps on the sex chromosomes promote genetic exchange across the species. Collectively these results suggest that ongoing sex-specific evolutionary processes, in addition to suppressed recombination, play an important role in governing the distribution of variation on the sex chromosomes as well as in C.
purpureus as a whole.
What does the phylogeography of a widespread moss species reveal? New insights from the Holarctic moss
Dicranum scoparium
Lang, A 1 , Stech, M 1
1 University Leiden, The Netherlands
Many moss species have a broad or even cosmopolitan distribution. Morphological homogeneity which may hide cryptic speciation, a haploid dominant life-cycle and frequent vegetative reproduction are key factors that may influence the amount of genetic diversity in moss species and the way genetic variability of populations is structured at different levels, from local to continental scales. Dicranum scoparium Hedw. is found across the
Holarctic, defined here as Europe and the non-tropical parts of Asia, Africa north of the Sahara, and North
America south to the Mexican desert region. The species displays two features of particular interest: (1) it is polymorphic and has been divided into multiple, often geographically restricted taxonomic entities, which facilitates the study of the spatial genetic structure in correlation with morphological trait variability across its range; and (2) it is dioecious with reduced-sized males sometimes growing on the female plants, a character that likely enhances sexual reproduction and reduce the relative importance of clonality. Because of its polymorphism, D. scoparium is often confused with taxonomically close species like D. bonjeanii , D. howellii , D. majus and D. japonicum . Here phylogenetic and phylogeographic relationships within D. scoparium s.l. are inferred based on five chloroplast DNA markers
( trn Ltrn F, trn Hpsb A, trn Trps 4, rpo B, rps 19rpl 2) and one nuclear marker (ITS), with further analyses based on
DNA fingerprinting planned as well. The extent to which the geographic distribution patterns, morphological trait diversity, and genetic diversity are linked, is investigated.
The results show very little intraspecific genetic variability with a number of haplotypes per marker ranging between five and eight. More surprising is the very low variability also at the interspecific level, which indicates that morphospecies cannot be molecularly distinguished using common DNA sequence markers.
Although biogeographic structuring of haplotype diversity is low, a continental disjunction between
Europe and Northern America for all chloroplast markers and a higher genetic variability in the latest continent are observed. The results will be discussed with respect to hypotheses of recent long-distance dispersal or ancient vicariance as suggested in previous studies on molecular evolution, molecular dating and measures of gene flow.
1
Regional and intercontinental phylogenetic patterns in peat mosses (
Sphagnum
)
Ricca, M 1 , Shaw, J 1
Duke Unniversity, Durham, USA
Like most mosses, Sphagnum species tend to have broad geographic distributions that span several continents. In addition to circumboreal species, some northern taxa have discontinuous disjunctive ranges including 'amphi-
Atlantic' and 'Pacific Rim' patterns. There have arguably been more genetic analyses of Sphagnum species than of any other bryophyte genus, revealing complex evolutionary and biogeographic patterns. We review case studies in which genetic data permit resolution of phylogeographic structure, including examples from eastern North America and Europe, northern South
America, and Australia–New Zealand–southern South
America. Estimates of divergence times between
European and North American populations of several species identify Quaternary divergence with limited (but nonzero) subsequent intercontinental migration. High levels of molecular similarity between plants on the two continents result from both retention of ancestral polymorphism and ongoing gene flow. Within eastern
North America, S. macrophyllum contains two divergent lineages, one distributed from New Jersey northward to
Newfoundland and the other southward from New Jersey to the Gulf coast. Genetic data indicate that the two are nearly or completely reproductively isolated. This northern-southern geographic pattern has been detected in several other bryophytes of eastern North America.
Recent work in the Southern Hemisphere shows that at least three peatmoss species are represented by both diploid and triploid gametophytes. In S. falcatulum , diploid gametophytes have only been detected in New
Zealand whereas triploids occur in New Zealand,
Tasmania, and Chile. In S. australe , diploid and triploid gametophytes co-occur at some New Zealand sites.
Sphagnum is increasingly being distinguished as a useful model for evolutionary and phylogeographic research, as these peatmosses have long been valued for research on community structure and ecosystem function.
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Implications of phylogenetic analyses of basal angiosperms and fossil seed plants for origin of the angiosperm flower
Doyle, JA 1
1 Dept of Evolution & Ecology, University of California,
USA
Elucidating the origin of the angiosperm flower involves two related problems: reconstructing the morphology of the flower in the most recent common ancestor of extant angiosperms, and identifying homologous structures in other seed plants. From a phylogenetic point of view, the first question can be approached by rooting the angiosperm tree and optimizing the evolution of morphological characters on it, while the second requires identification of the closest outgroups of angiosperms and interpretation of the morphology of their reproductive structures. Great progress on first problem has come from molecular analyses of living angiosperms, complemented by studies of Cretaceous fossil flowers.
Molecular analyses consistently identify either
Amborella or Amborella plus Nymphaeales as the sister group of all other angiosperms. Both rootings imply that the ancestral flower had numerous undifferentiated tepals and stamens, which may have been either spiral or trimerous, and several ascidiate carpels, with a single bitegmic ovule. It may have been either unisexual or bisexual, but the presence of abortive stamens in female flowers of Amborella would support the latter. Although there have been suggestions that basal angiosperms with extremely simple flowers, such as Hydatellaceae,
Chloranthaceae, and the Early Cretaceous fossil
Archaefructus , represent a pre-floral state, it is more parsimonious to interpret these as derived from flowers with more numerous parts. Progress on identification of angiosperm outgroups has been more elusive, because of homoplasy and difficulties in homology assessment in morphological characters and conflicts between morphological and molecular data. Most morphological analyses have associated angiosperms, Gnetales, and fossil Bennettitales and Pentoxylon in an anthophyte clade, but they disagree on whether this clade is related to coniferophytes or to 'seed ferns' such as corystosperms, glossopterids, and Caytonia . In contrast, molecular analyses of living seed plants support either trees with Gnetales nested in conifers and angiosperms sister to other living seed plants, or trees with Gnetales sister to other seed plants; the former trees are favored by likelihood-based analyses. However, a morphological analysis including recently recognized conifer-like features of Gnetales found that anthophyte trees and trees with Gnetales in conifers were equally parsimonious. In the latter trees, angiosperms were linked with glossopterids, Pentoxylon , Bennettitales, and Caytonia .
This is consistent with homology of the angiosperm bitegmic ovule with the cupules of glossopterids and
Caytonia , but it is unclear whether the carpel was derived from the rachis of a cupule-bearing sporophyll or from a leaf with an axillary branch bearing one or more cupules; the latter might provide a better prototype for a
264 uniovulate ascidiate carpel. Stamens may represent similar units bearing two microsynangia. If Bennettitales are also related to angiosperms, their very different ovulate structures pose problems for both scenarios.
However, there is currently no consensus that any known fossil taxon is a stem relative of angiosperms. This underlines the need for more data on the morphology of known fossil taxa, discovery of closer angiosperm outgroups, and better information on homologies of angiosperm structures based on evo-devo studies.
Current highlights and problems in evolutionary flower morphology
Endress, PK 1
1 Institute of Systematic Botany, University of Zurich,
Switzerland
In light of the current molecular phylogenetic revolution in angiosperm systematics our approach to floral morphological features needs to be reassessed. This reassessment is twofold. Firstly we need to comparatively study clades that have been newly esablished by molecular phylogenetic studies. And secondly, and complementary to the first, we need to study why systematists were misled in the recognition of suprafamilial and supraordinal clades in the premolecular era. Here I will concentrate on the second approach. I focus on some of the premolecular angiosperm orders that have been profoundly disassembled and reassembled in the past twenty years. Using the classification by A.
Cronquist as an example, I will select some plurifamilial orders that have considerably changed in their familial composition. Such former problematic orders occur mainly in the basal angiosperms and in rosids, whereas asterids and monocots have been less affected; specifically they include Magnoliales, Laurales,
Nymphaeales, Hamamelidales, Dilleniales, Theales,
Violales, Rosales, Rafflesiales, Celastrales, Euphorbiales,
Polygalales, Sapindales, Geraniales, Solanales, Lamiales, and Liliales. I will explore what floral features may have been used to hold them together initially, and what different and new features are more uniform in the clades of the new ordinal circumscriptions. Some features of floral architecture apparently evolved many times in eudicots and are present in quite disparate clades, such as
(ob)diplostemony, polystemony, fusion of petals and stamens, features of a buzz pollination syndrome, and features used in alpha-taxonomy to describe gynoecia, e.g., ovary position and placentation patterns. In contrast, it will be shown that morphological features of the ovules are powerful markers in high-level systematics but have been neglected and underestimated in earlier classifications.
Novel insights and future directions for understanding the developmental genetic basis of convergent floral trait evolution
Hileman, L 1 , Preston, J 1
1 University of Kansas, Lawrence, USA
Repeated (convergent) evolution of similar floral traits, such as unisexuality, perianth reduction associated with wind-pollination, bilateral symmetry, and presence of

nectar spurs, has been pervasive during angiosperm diversification. The repeated occurrence of a particular trait is considered evidence for adaptive evolution, and convergence of many floral traits likely results from natural selection associated with particular pollination and/or mating strategies. Patterns of convergent evolution in flower traits are increasingly well documented through detailed and data-rich angiosperm phylogenetic analyses. However, whether the repeated evolution of floral traits has occurred through parallel recruitment of the same, or similar, developmental genetic programs, remains a largely unanswered question. The field of evolutionary development is making significant progress in determining the developmental genetic basis for repeated evolution of floral bilateral symmetry. Strikingly, data derived mainly from representative core-eudicot taxa implicate the independent (parallel) recruitment of a CYCLOIDEA
(CYC)-dependent developmental genetic pathway in the multiple origins of flower bilateral symmetry, and have raised further critical questions regarding the evolution of flower symmetry. Is the CYC-dependent floral symmetry genetic pathway restricted to eudicots? What are the developmental roles for CYC-like genes in flowering plant species with radially symmetrical flowers? What properties of a CYC-dependent pathway might result in its repeated recruitment to specify a similar trait? We will provide data evidencing evolution of the CYCdependent pathway in the development of floral bilateral symmetry outside eudicot angiosperms, as well as in the evolutionary reversal to floral actinomorphy.
Additionally, we will discuss future research directions aimed to enhance our understanding of how and why this pathway has been repeatedly recruited to specify bilateral flower symmetry during angiosperm diversification.
Patterns of floral development and evolution: examples from the Ericales
Schönenberger, Jürg 1 , von Balthazar, M 2
1 Dept of Structural and Functional Botany, University of
Vienna, Austria; 2 Dept of Systematic and Evolutionary
Botany, University of Vienna, Austria
The asterid order Ericales comprises about six per cent of the total eudicot species diversity and is highly diverse at all levels of structural and functional biology, particularly so also in the flowers. In many respects, floral structure in the order is transitional between the rosids and the euasterids with features such as organ number
(oligomery/polymery), floral symmetry (poly-
/monosymmetry), petal union (chori-/sympetaly), synorganization of corolla and androecium
(presence/absence of a stamen-corolla tube), and the number of integuments (one/two) being distinctly homoplasious. In recent years, Ericales have found considerable interest among molecular systematists as well as morphologists. As a result, interfamilial phylogenetic relationships are fairly well understood and at least for some of the subclades, floral structure and diversity are well documented. These recent advances make it possible to study character evolution in much greater detail than before. In this paper, we focus on patterns of floral development and bring together new unpublished data and results from earlier ontogenetic studies. We summarize developmental data for patterns of floral phyllotaxis, symmetry, sympetaly, and merosity.
Special emphasis is given to the development of polystemony, which is present in at least twelve of the 22 currently recognized ericalean families. Reconstruction of character evolution indicates that polystemony has apparently evolved several times within the order, both from haplo- and diplostemonous ancestors. The repeated, independent evolution of polystemony is reflected in a high diversity of early developmental patterns including special features such as ring meristems, stamens fascicles, stamen pairs, centripetal as well as centrifugal stamen initiation. It appears that in Ericales many floral traits at the organizational level are evolutionary labile.
This is in stark contrast to the euasterids, where floral organization is much more stable and evolutionary labile floral traits occur almost exclusively at the level of floral construction and pollination syndromes. We argue that the study of the genetic basis of the evolutionary lability of ericalean floral traits might be a powerful approach to address questions about floral evolution in the asterids as a whole.
Floral symmetry and angiosperm diversification: state-of-the-art and new approaches
, Nadot, S 1
1
Sauquet, H 1
Univ. Paris-Sud, France
Flowers display an incredible variation of shape, structure, and colour throughout the angiosperms. This diversity has long been thought to be closely linked to the large number (>260,000) of extant species of flowering plants with respect to other land plants.
However, patterns of species richness vary widely within the angiosperms, suggesting that some traits may have promoted significantly the diversification of particular clades. One such key innovation is floral bilateral symmetry (zygomorphy), which has evolved repeatedly from radial symmetry (actinomorphy) and is often associated with specialized insect pollination. Various approaches have been used to address the evolution of zygomorphy across the angiosperms, including phylogenetic comparative methods and ancestral character reconstruction. Whereas some studies have used supraspecific terminals to represent floral symmetry in different clades, others have opted for the use of exemplar species (although not yet at the scale of angiosperms as a whole). However, very few higherlevel studies have distinguished among the various conditions of floral symmetry, which may affect all or only some of the floral organ series (i.e., the perianth, the androecium, and/or the gynoecium), involve different developmental processes (e.g., differential growth or specialization, fusion, or suppression of parts), and be expressed along different axes (dorsoventral, transverse, or oblique). Here, we review previous work aimed at reconstructing patterns of floral symmetry across the angiosperms and propose a new approach implemented in an innovative database framework (PROTEUS). This approach is based on exemplar species sampled from all currently recognized families of flowering plants and the comparison of multiple expressions of the floral symmetry character. Using a dated supertree that best summarizes currently understood relationships and divergence times among angiosperm families, we present comparative analyses aimed to: 1) identify other floral
265

traits possibly correlated with floral symmetry, in particular stamen number and perianth merism; 2) test whether particular forms of zygomorphy have promoted diversification rates more than others. Our results provide an important framework to help guiding the choice of species for finer-scale Evo-Devo studies, targeted at understanding the different genetic pathways in which zygomorphy and other key floral traits have evolved.
Floral evolution in animal-pollinated Australian angiosperm clades: patterns and potential explanations
Weston, PH 1 , Wilson, PG 1 , Conn, BJ 1 , Rymer, PD 1
1 National Herbarium of New South Wales, Sydney,
Australia
The Australian continent became a distinct biogeographic entity when its last land connection with Antarctica–
South America was severed in the late Eocene. Its biota has thus evolved for most of the past 35 million years in isolation, apart from sporadic immigration across significant water gaps by long distance dispersers. This has left Australia with a diverse, highly endemic biota that also lacks some ecologically influential clades that are widespread elsewhere. For example, although
Australia has a diverse bee fauna, it has no native species of the widespread genera Apis and Bombus . Numerous, speciose, endemic subclades in the angiosperm families
Myrtaceae, Fabaceae and Proteaceae dominate a large percentage of the land surface. The flowers of these plants are pollinated by a diverse fauna of bees and other hymenopterans, flies, beetles, lepidopterans and nectar- and pollen-feeding birds, bats and marsupials. Late
Cenozoic evolution of floral traits in the animalpollinated Australian angiosperm flora will be reviewed in the light of generic-level phylogenetic reconstructions for the dominant clades listed above as well as other predominantly animal-pollinated clades. The following floral characters (where known) will be mapped onto these phylogenies using parsimony and likelihood criteria: symmetry, mery, degree of connation or cohesion of floral whorls, adnation between floral whorls, colour of floral whorls, aestivation, nectary presence/absence, degree of stamen and style exsertion, stamen number, kind of anther dehiscence, ovary position and visually dominant floral part(s). Broadly circumscribed functional pollinator groups (where known) will also be mapped onto the plant phylogenies.
Preliminary analyses suggest that some patterns that were recognized in the nineteenth century as pollination syndromes on the basis of non-Australian examples are represented in the Australian flora by correlated phylogenetic changes in floral form and function. For example, the evolution of bird pollination is frequently accompanied by the evolution of red floral parts, connate perianth segments and an exserted style. Are the patterns and processes of floral evolution in Australia as unique and distinct as its flora? The short answer to this question is 'no'. Most of them seem to have analogues elsewhere.
For example, despite the absence of native bumble bees, porose anther dehiscence has still evolved multiple times in the Australian flora in plants that are pollinated by other bee taxa that are also capable of sonication.
However, some patterns and processes in floral evolution are more prevalent in Australia than elsewhere. Take the
266 evolution of sexual deception in Australian orchids, for example, which has evolved independently in at least six different lineages in the tribe Diurideae alone. Compare this with its similarly speciose, predominantly
American–Asian sister group, the tribe Cranichideae, in which the only known sexually deceptive clade is the
Australasian genus Pterostylis . At least one macroevolutionary pattern seems peculiarly Australian: most of the continent is dominated by two huge, generalist-pollinated angiosperm clades, the eucalypts and Acacia , which have flowers dominated by exserted stamens.
Comparative floral structure and systematics in
Ochnaceae s.l. (Ochnaceae, Quiinaceae,
1
Medusagynaceae), Malpighiales
Matthews, ML 1 , do Carmo E. Amaral, M 2 , Endress, PK 1
Institute of Systematic Botany, University of Zurich,
Zurich, Switzerland;
Campinas, Brazil
2 Institute of Biology, UNICAMP,
The revolution in biological phylogenetics over the last
15–20 years by the introduction of molecular techniques has led to a profound reorganization of the systematics of organisms, which in the flowering plants is reflected by the continuously improved supraordinal and ordinal classifications. Thus we are now in the paradoxical situation that new knowledge on systematic relationships is available, yet the structural and biological features that characterize these new clades are unknown due to a lack of comparative studies. Such studies are essential, not only for a better understanding of organisms, but also for the evaluation of the systematic position of fossils and for evolutionary developmental (evo-devo) studies on flowers. Rosids represent one of the largest eudicot clades in terms of higher level systematics (17 orders).
Many orders are relatively well supported. However, relationships between orders or between families within orders are in many cases still unresolved. The COM
(Celastrales–Oxalidales–Malpighiales) clade represents one such problematic group, and Malpighiales (with over
40 currently recognized families) represent the largest and most recalcitrant order within angiosperms. In two previous studies on Malpighiales we characterised the well supported suprafamilial clades, Chrysobalanaceae s.l. and Rhizophoraceae s.l. (plus some additional families), using comparative floral structural studies and identified a number of potential synapomorphies for these clades. In this study our focus is Ochnaceae s.l., another well supported clade, which includes the larger family Ochnaceae and the small families Quiinaceae and
Medusagynaceae. A literature search has thus far revealed a few common plus some unusual shared, more general floral features such as a polystemonous androecium and the tendency of the gynoecia to have more than 5 carpels (largest numbers recorded: 15 in
Ochnaceae, 13 in Quiinaceae, and 23 in
Medusagynaceae). Quiinaceae and Medusagynaceae share partly unisexual flowers, separate stylar branches with capitate stigmas, a pronounced ovary roof, and often
2 ovules per carpel. In contrast, Ochnaceae with largely bisexual buzz-pollinated pollen flowers (often monosymmetric) have unified styles and generally pointed stigmas, and their ovaries show a broad range of placenta forms and ovule numbers. To gain a more in-

depth understanding of the seemingly disparate structure of the flowers of these families a detailed study of both their external and internal morphology is necessary.
Flowers from genera of the three families are comparatively studied using a combination of serial microtome sections of floral buds and anthetic flowers plus scanning electron microscopy (SEM). Results of these studies including potential synapomorphies for this clade and comments on the occurrence of floral features within both Malpighiales and the COM clade will be given.
What we know and don’t know about the 'Afunction': the role of AP1/FUL genes in basal eudicot flowers
Pabon Mora, N 1 , Litt, A 2
1 New York Botanical Garden, City University of New
York, USA; 2 New York Botanical Garden, USA
According to the ABC model of flower development, the
A-class genes are responsible for specifying floral meristem as well and sepal and petal identity. One of these A-function genes in Arabidopsis is APETALA1
(AP1). AP1 belongs to a lineage of MADS-box transcription factors unique to flowering plants called the
APETALA1/FRUITFULL gene lineage. This lineage, like others involved in flower development, has undergone several duplications in different groups of angiosperms. Gene duplication occurs frequently in plants and has been postulated as a potential source of morphological variation due to the expansion and functional diversification of gene families. The AP1/FUL lineage provides an opportunity to explore how duplication events have resulted in new gene functions or combinations of functions, and the commensurate increase in morphological diversity and complexity. In the AP1/FUL gene lineage a major duplication event coincided with the origin of the core eudicots and produced the euFUL and the euAP1 clades. Interestingly, genes belonging to each clade are associated with different functions. Other euAP1 genes in core eudicots have been shown to be involved in the transition to the floral meristem and sepal identity like AP1, but do not play a role in petal identity. Evidence suggests the canonical A-function in flower development is a
Brassicaceae-specific phenomenon, attributed possibly to unique additional local duplications. On the other hand
FRUITFULL (FUL) in Arabidopsis functions redundantly with AP1 during floral meristem specification but plays a distinct role in fruit development and determinacy. Other euFUL genes function similarly to FUL suggesting that differences in the functions of
AP1/FUL core eudicot paralogs might have arisen with the duplication event. Assessing functional evolution in this gene lineage requires the characterization of genes in species that diverged before the duplication event.
Functionally, these pre-duplication FUL-like genes have scarcely been explored; the predicted proteins show sequence similarity to euFUL proteins, and evidence suggests that in temperate grasses they play a role in the transition to reproduction after vernalization. In order to characterize the plesiomorphic role of the gene lineage and to better understand the role of gene duplication in the functional evolution of the AP1/FUL lineage, we explored the expression and function of FUL-like genes in Papaver somniferum (opium poppy) and Eschscholzia californica (California poppy), both Papaveraceae members, within the basal eudicots. In order to investigate the endogenous role of FUL-like genes we used Virus Induced Gene Silencing (VIGS). Downregulated lines, in both species, show increased branching, homeotic conversion of sepals into leaf-like organs and fruit defects. These data shows that FUL-like genes control axillary meristem growth, floral meristem and sepal identity and play a key role in fruit development. Because the FUL-like gene functional repertoire encompasses essentially all roles previously described for euAP1 and euFUL genes we postulate subfunctionalization as the driving force contributing to diversification in the AP1/FUL gene lineage as a result of the core-eudicot duplication event. In this context we discuss the universality of 'A-function' across angiosperms and explore putative changes in the protein networks before and after the gene duplication event.
1
Separating the sexes: dioecious floral evolution in
Plagianthus
(Malvaceae)
Skema, C 1 , Tate, JA 1
Institute of Molecular Biosciences, Massey University,
New Zealand
Dioecy, although proportionally rare, has evolved repeatedly throughout the angiosperms. The study of the floral evolution, development, genetics and ecology of these dioecious taxa provides an invaluable view into the control of gender expression in flowering plants and the selection pressures for maintaining distinct male and female individuals. Yet, only a few classic examples
(e.g., Silene ) have been well studied in all these aspects.
The five genera within the Plagianthus alliance of tribe
Malveae, Malvaceae ( Asterotrichion , Gynatrix , Hoheria ,
Lawrencia , Plagianthus ) provide a unique opportunity to study the separation of the sexes given a remarkably high frequency of dioecy and the potential for its repeated evolution within the group. An in-depth study of the floral evolution, development and genetics within the alliance has begun with Plagianthus , a genus that contains two strictly dioecious species that are both endemic to New Zealand. To characterize its floral development, histological and scanning electron microscopy work was conducted on specimens of both species of Plagianthus collected from floral inception through to anthesis. Early floral developmental stages are identical between the two sexes with each developing the following in succession: five sepal primordia, a single ring-like wall of the corolla-androecium primordium that subdivides into five lateral petal primordia and 10–20
(sub)apical anther primordia, and finally the gynoecial primordium. Only in later development do the sexes differentiate, each arresting the development of the whorl of the opposite gender to produce functionally unisexual flowers, with the female flowers smaller than the males.
Externally, the androecium of a female flower differs from a male androecium in that it is smaller overall and the anthers are indehiscent and deflated in appearance.
This last characteristic is due to degradation of the anther contents in the final phase of female androecial maturation. At the time of degeneration, the majority of cells appear to be microsporocytes, implying that development usually arrests prior to microsporogenesis.
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A very small minority of these cells have been interpreted as tetrads, although meiosis has yet to be observed within female anthers. The gynoecium of male flowers is roughly one-third the size of a female gynoecium and develops a non-functional ovule. The male stigma remains hidden within the androecial tube at anthesis and has substantially less surface area than the prominently exserted stigma found in female flowers. An investigation into the genetic differences underlying gender expression is underway by means of a comparison of 454 sequence data of Plagianthus floral cDNA libraries of males versus females.
X-ray tomography of plant tissue: staining methods allow 3D imaging with high contrast and high resolution
Staedler, YM 1 , Schönenberger, J 1
1 Dept of Structural and Functional Botany, University of
Vienna, Austria
Study of plant morphology, development and function requires detailed three-dimensional visualization and modeling. X-ray micro-computed-tomography (micro-
CT) allows straightforward visualization and modeling of complex 3d objects. However, the low absorbance of most plant tissues has earlier been an obstacle for the use of this method in plant sciences (with the notable exception of permineralized or charcoalified fossils and wood samples). Here we present several staining methods that allow high contrast and high resolution reconstructions of any plant tissue/organ using a commercial micro-CT system. We selected a broad range of stains from the Transmission Electron Microscopy literature and have tested their usefulness in X-ray tomography. Stains included non-metals such as Iodine and a selection of light and heavy transition element compounds. Different concentrations of staining solutions were tested, as well as different infiltration times. We studied staining effect on material of open flowers, flower buds with thick calyptras, and isolated carpels (FAA and ethanol preserved). These varying objects allowed us to quantify: (1) contrast improvement,
(2) homogeneity of the stain, (3) penetration of the tissue, and (4) selectivity of stain. The stains proved unequal in their performance. If (1) all stains allowed for improvement in contrast, improvement was however especially noticeable with metallic stains. (2)
Homogeneous staining was obtained with most of the stains on small samples, but only with few on large samples. (3) Good penetration of the tissue was very unequal and, when assessed, highly dependent on solvent composition. (4) Highly selective staining as such was not obtained. However, noticeable staining was obtained, with varying compounds, for pollen, stigmata, ovules, vasculature, cell walls, and cell nuclei. Use of the appropriate stains allows for good contrasting and visualization of all probes studied so far.
1
Plant functional traits: adding in the flowers for
Australian Proteaceae
Zanne, A 1 , Miller, Eliot 2
University of Missouri – St Louis, USA; 2 Macquarie
University, Sydney, Australia
268
Plants exhibit an enormous variation in how they are constructed and how they function. Theses different constructions determine the environmental settings where plants can survive and reproduce. Recent work has identified a series of plant functional traits representing differences in species strategies. These studies have focused on vegetative and dispersal aspects of plants but have yet to incorporate floral traits. Important axes of plant functional trait variation have been identified, including the leaf economics spectrum, leaf-twig size deployment, seed size, plant height, and wood anatomy.
While identifiable patterns of trait coordination across species and association of those patterns with particular environments have been found, high variation within sites suggests that many different plant constructions can work at a given site. Floral displays mediate the pollen delivery among plants that ensure fertilization and passing on of a plant’s genes. Thus floral traits should be an important aspect of how plants function and may show different patterns of variation (both within and across species) than vegetative functional traits. In this study, we contrast the intraspecific variation in and evolutionary lability of vegetative and floral traits across
Proteaceae from eastern Australia. The Proteaceae represent a dominant clade in Australia found in most regions in the continent. It shows high variation in functional traits (e.g., tiny needle like to large leaves and short shrubs to tall trees), as well as pollinator strategies
(e.g., insects to birds), and thus should capture a wide range of variation in plant trait axes. We test the following three expectations. 1. Due to the strong selection for optimal pollen placement by pollinators but many ways that plants can acquire light, nutrients and water, there is lower within species variation in floral than vegetative functional traits. 2. To minimize hybridizations and/or pollen discounting, floral traits show little phylogenetic signal when compared to vegetative traits. 3. Strong coordination is found for a suite of traits that relate to the pollinators (e.g., insect versus bird pollination). We suggest floral traits are an important dimension of plant trait variation to consider in studies of plant functional diversity.
Can GrassBase grow into an international data repository for the evolving classification of grasses?
Vorontsova, MS 1
1 Royal Botanic Gardens, Kew, UK
Classification updates from the grass phylogenetics boom are scattered across numerous publications. The lack of any centralised compilation means classification changes are not efficiently communicated or implemented. GrassBase is a commonly consulted but old fashioned reference source, a standardized compilation of nomenclature, synonymy, and typification of ca. 64,000 names and coded morphological descriptions of ca. 12,000 species and genera recorded in
1,100 DELTA characters. This database has been created by Derek Clayton over 20 years of data entry from IPNI,

published floras, and monographic revisions. GrassBase currently uses the morphological classification system based on Genera Graminum with no links to the Grass
Phylogeny Working Group classification system. This talk will describe the history of Grassbase, its current implementation, mechanisms for incorporating both classification systems into Grassbase, and opening
GrassBase for worldwide contributions. Moving to a common ownership multi-author model could allow
GrassBase to become the centralised repository for all grass taxonomic and descriptive data. Linking the new classifications to existing species descriptions, nomenclature, and typification would create an unprecedented online resource – an evolving World Flora of Grasses.
A multi-gene study of relationships within the
Australian grass genus
Austrostipa
Syme, A 1 , Udovicic, F 1
1 Royal Botanic Gardens Melbourne, Australia
The Australian endemic grass genus Austrostipa
S.W.L.Jacobs & J.Everett plays an important role in ecological habitats and pastures, and accordingly its species have been the subject of recent studies in seed germination, physiology and revegetation. Particular species are irritants to grazing animals or are ecologically important to degraded habitats, and it is therefore useful to know their closest relatives within the genus. Although the genus is speciose, abundant, and ecologically significant, the sub-generic classification of its 62 species has not been comprehensively tested with molecular data.
We used three molecular markers to infer a phylogeny to better understand whether species in the thirteen subgenera formed natural monophyletic clades. In 1996 the genus was split from Stipa and thirteen subgenera were formally described. Since then, three molecular studies investigated relationships in Austrostipa , finding poor support for almost all of the current subgenera.
These studies used the nuclear internal transcribed spacer
(ITS) region and inferred phylogenies using parsimony and clustering methods. In the present study, we sampled comprehensively across the genus (> 80% of species) using the chloroplast genes ribulose bisphosphate carboxylase/oxygenase large subunit ( rbc L) and maturase K ( mat K), as well as the nuclear ITS region.
We used phylogenetic inference methods based on
Bayesian likelihood, which can include more probable evolutionary models of nucleotide change and can be incorporated into large treesearch algorithms. The resulting molecular phylogeny of Austrostipa shows which subgeneric clades are supported by current evidence and provides the most comprehensive phylogenetic framework for ecological, physiological and genetic studies on the genus.
Phylogenetic relationships and changing circumscriptions in the large PPAM clade (poaceae:
1 poeae: subtribes Alopecurinae, Milliinae, Phleinae,
Poinae, and Puccinelliinae)
Gillespie, L 1 , Soreng, R 2
2
Canadian Musuem of Nature, Ottawa, Canada;
Smithsonian Institution, Washington, DC, USA
The grass subtribes Alopecurinae, Miliinae, Phleinae,
Poinae, and Puccinelliinae form a strongly supported clade (PPAM) within the grass tribe Poeae s.l. The
PPAM clade comprises ca. 750 species distributed worldwide in cool temperate and tropical alpine regions.
We used parsimony and Bayesian analyses of plastid trn Ttrn Ltrn F and nuclear ribosomal ITS and ETS sequence data to infer phylogenetic relationships and assess generic and tribal circumscriptions in the PPAM lineage. Only Puccinellinae and Phleinae (with only one of its 2 putative genera sampled) formed strongly supported clades. Poinae and Alopecurinae are paraphyletic or polyphyletic as currently circumscribed, with Poa forming a clade separate from a clade of all other Poinae genera plus all Alopecurinae genera sampled. The PPAM clade includes numerous genera traditionally allied with tribe Aveneae (now included in
Poeae s.l.), including all sampled genera of subtribes
Alopecurinae, Phleinae and Cinninae (the latter now in
Poinae), six Poinae genera ( Apera , Dissanthelium ,
Gaudiniopsis , Simplicia , Tovarochloa , Ventenata ), and the type species of Avenula . The PPAM clade also includes genera previously allied with tribes Stipeae
( Milium ) and Meliceae ( Anthochloa ). Many changes in generic circumscription have been made recently in subtribe Poinae as a result of our molecular studies.
Anthochloa , Aphanelytrum , Austrofestuca s.s.,
Dissanthelium , Eremopoa , Neuropoa , Parodiochloa ,
Tovarochloa , and Tzvelevia are now included with Poa or considered to belong here and are pending revision. To accommodate species previously placed in Poa, but which fall outside of the core Poa clade in Poinae, we recently described the new genera Nicoraepoa , Sylvipoa , and Saxipoa , and resurrected Arctopoa as a genus,
Further major changes in the circumscription of PPAM subtribes are suggested by our current data, including the placements of Coleanthus , Colpodium , and Poa hayachinensis , the latter likely representing a new Poinae genus.
Phylogenetic structure in the large genus
Poa
4
3
2
1
(Poaceae: Pooideae: Poeae)
Soreng, RJ 1 , Gillespie, LJ 2 , Giussani, LM 3
MA 3 , Negritto, MA 4
, Scataglini,
Smithsonian Institution, Washington DC, USA;
Canadian Museum of Nature, Ottawa, Canada;
Instituto de Botanica Darwinion, San Isidro, Argentina;
Universidad de Concepcion, Concepcion, Chile
A phylogenetic analysis of Poa L. (over 500 spp.) using an expanded sample of species from around the world
(269 spp. and an additional 39 infraspecies have been examined for plastid and or nrDNA types), with more complete sampling for plastid trn TLF region and nrDNA spacer regions ITS and ETS than previous studies, revealed two large derived clades (number of species based on morphology) subgen. Poa ( Poa –
Homalopoa ; 360 spp.) and subgen.
Stenopoa (85 spp.) that contain most of the species in the genus. The subgen.
Ochlopoa clade contains a further 36 spp., subgen.
Pseudopoa ( Eremopoa ) 5 spp., subgen. Sylvestres 8 spp., and sect. Pandemos 2 spp. The small 'N-clade' had 3 diploid species confined to it by plastid and nrDNA, including the monotypic sect. Nanopoa and two species placed in other non-monophyletic sections, and 11
269

polyploid species (including all species of sect.
Secundae , and some of Glariosae ) that share plastid types with it, but ITS–ETS types with the Stenopoa clade. Individuals of Poa bulbosa (sect. Arenariae ) had plastid/nrDNA combinations either aligning within the N clade, or within the clade with consectional species in the
Ochlopoa clade, but no intermixed types. An odd and strongly supported nrDNA ITS–ETS 'X-clade', for which no corresponding plastid clade has been identified, resolved for 40 species (no diploids known among them) from around the Pacific basin (China, Hawaii, Japan,
New Guinea, New Zealand, North and South America: including sects. Acroleucae , Malacanthae ,
Siphonocoleus , and elements of Orienos , Pauciflorae , and about 2/3 of New Zealand species). Plastid sequences of species with X-type nrDNA mostly resolved in the Poa – Homalopoa clade, but 4 other species aligned with sects. Orienos and Pauciflorae .
Based on DNA studies Poa is considered to include the genera Anthochloa , Aphanelytrum , Austrofestuca s.s.,
Dissanthelium , Eremopoa , Parodiochloa , Tovarochloa ,
Tzvelevia ; exclude the more recently described genera
Arctopoa , Nicoraepoa , Saxipoa , Sylvipoa ; and to have reticulate relationships with Aniselytron , Arctopoa , and
Nicoraepoa .
Plastid-based phylogenetics of Agrostidinae,
Koeleriinae and allied subtribes in the ‘Aveneae-type plastid DNA clade’ of the grass tribe Poeae (Poaceae:
Pooideae)
Saarela, JM 1 , Peterson, PM 2 , Soreng, RJ 2 , Paszko, B 3
1 Canadian Museum of Nature, Ontario, Canada; 2 Dept of Botany, National Museum of Natural History,
Smithsonian Institution, Washington DC, USA; 3 W Szafer
Institute of Botany, Polish Academy of Sciences, Poland
The ‘Aveneae-type plastid DNA clade’ of tribe Poeae is a major, species-rich lineage of cool-season grasses that includes more than 40 genera, including the large, broadly distributed and taxonomically difficult genera,
Agrostis , Calamagrostis (including Deyeuxia ) and
Trisetum . Five or six subtribes (Agrostidinae, Aveninae,
Brizinae, Koeleriinae (sometimes), Phalaridinae,
Torreyochloinae) are currently recognized in the clade, but multiple aspects of molecular relationship among and within these hypothetical 'lineages' remain largely equivocal, or conflict with current circumscriptions. The status of some smaller segregate genera is uncertain, and several have not yet been included in molecular analyses.
In existing phylogenetic trees the North American genera
Graphephorum , Peyritschia , and Sphenopholis are intermixed with some North American species of Trisetum and Mexican species of Calamagrostis , but most species of Calamagrostis from South America and Eurasia and
Trisetum from South America have not yet been sampled. Generic limits within Agrostidinae are not clear, particularly in plastid-based phylogenies, which so far have been based on limited data: for example,
Ammophila is intermixed with species of Calamagrostis , deep lineages in Calamagrostis have not been identified, and species of Agrostis , Lachnagrostis , and Polypogon are, for the most part, intermixed. Here, we continue exploration of the evolutionary history in and among these lineages by expanding the taxon and plastid sampling of our earlier work, including previously un-
270 sampled species of Calamagrostis / Deyeuxia and
Trisetum from South America and Eurasia, and new data from multiple plastid regions, including mat K, psb Atrn H, psb Kpsb I, and atp F-H. We also explore the utility of plastid sequence data for DNA barcoding in the
‘Aveneae-type plastid DNA clade’ of tribe Poeae.
Phylogeny of the Chloridoideae (Poaceae) based on analysis of 11 DNA loci
Peterson, PM 1 , Romaschenko, K 2 , Johnson, G 1
1 Smithsonian Institution, Washington DC, USA; 2
Institute of Barcelona, Spain
Botanic
We conducted a molecular phylogenetic study of the subfamily Chloridoideae using nine plastid DNA sequences ( ccs A, mat K, ndh A intron, ndh F, rpo C2, rps 16trn K, rps 16 intron, rps 3, and rpl 32-trnL) and two nuclear sequences (ITS and At103). Our large original data set includes more than 300 species (21.1%) representing 105 genera (74%) of the grasses currently placed in the Chloridoideae. The maximum likelihood and Bayesian analysis of DNA sequences provides strong support for the monophyly of the Chloridoideae; followed by, in order of divergence: a Centropodieae clade with Centropodia sister to an unnamed genus with two species; a Triraphideae clade with Neyraudia sister to Triraphis ; an Eragrostideae clade with the Cotteinae
(includes Cottea and Enneapogon ) sister to the Uniolinae
(includes Entoplocamia , Tetrachne , and Uniola ), and a terminal Eragrostidinae clade of Ectrosia , Harpachne , and Psammagrostis embedded in a polyphyletic
Eragrostis ; a Zoysieae clade with Urochondra sister to a
Zoysiinae ( Zoysia ) clade, and a terminal Sporobolinae clade that includes Spartina , Calamovilfa , Pogoneura , and Crypsis embedded in a polyphyletic Sporobolus ; and a very large terminal Cynodonteae clade that includes 13 monophyletic subtribes. The Cynodonteae includes, in alphabetical order: Aeluropodinae ( Aeluropus );
Boutelouinae (Bouteloua); Eleusininae (includes
Apochiton , Astrebla with Schoenefeldia embedded,
Austrochloris , Brachyachne , Chloris , Cynodon with
Brachyachne embedded in part, Eleusine , Enteropogon with Eustachys embedded in part, Eustachys ,
Chrysochloa , Coelachyrum , Leptochloa with Dinebra embedded, Lepturus , Lintonia , Microchloa , Saugetia ,
Schoenefeldia , Sclerodactylon , Tetrapogon , and
Trichloris ); Hilariinae ( Hilaria ); Monanthochloinae
(includes Distichlis , Monanthochloe , and Reederochloa );
Muhlenbergiinae ( Muhlenbergia with Aegopogon ,
Bealia , Blepharoneuron , Chaboissaea , Lycurus ,
Pereilema , Redfieldia , Schaffnerella , and Schedonnardus all embedded); Orcuttiinae (includes Orcuttia and
Tuctoria ); Pappophorinae (includes Neesiochloa and
Pappophorum ); Scleropogoninae (includes
Blepharidachne , Dasyochloa , Erioneuron , Munroa ,
Scleropogon , and Swallenia ); Traginae ( Tragus with
Monelytrum , Polevansia , and Willkommia all embedded);
Tridentinae (includes Gouinia , Tridens , Triplasis , and
Vaseyochloa); Triodiinae ( Triodia ); and theTripogoninae
( Melanocenchris and Tripogon with Eragrostiella embedded). Based on our phylogenetic treatment the following 14 genera are polyphyletic: Astrebla ,
Brachyachne , Chloris , Cynodon , Enteropogon ,
Eragrostis, Eustachys, Leptochloa, Perotis, Sporobolus,
Tetrapogon, Tragus, Tripogon , and Willkommia ; and the

following 26 genera with two or more species were always portrayed as monophyletic: Aeluropus,
Blepharidachne, Bouteloua, Centropodia, Cleistogenes,
Ctenium, Dactyloctenium, Dignathia, Distichlis s.l
.,
Eragrostiella, Erioneuron, Lepturus, Melanocenchris,
Microchloa, Mosdenia, Muhlenbergia s.l
., Munroa,
Orcuttia, Orinus, Trichloris, Trichoneura, Triodia,
Triraphis, Tuctoria, Uniola , and Zoysia . In our study the
Cynodonteae still include 19 genera and the Zoysieae include a single genus that are not yet placed in a subtribe. Our phylograms indicate that the Chloridoideae might have originated in Africa and/or Asia since the basal lineages, tribes Centropodieae and Triraphideae, include species with both African and Asian distributions. Initial estimates of divergence times based on analysis of ndh F sequences indicate the Eragrostideae is the oldest lineage at 29.8 mya, followed by the
Cynodonteae at 27.5 mya, then the Centropodieae at
24.1 mya., the Zoysieae at 22.8 mya, and the
Triraphideae (youngest) at 10.4 mya.
Eocene caldera floras of Patagonia, Argentina: the peak of Australasian rainforest signal in South
1
America
Wilf, P 1 , Carvalho, M 1
Cuneo, R 3 , Johnson, K 4
, Hermsen, E 2 , Gandolfo, M 2 ,
2
Pennsylvania State University, University Park, USA;
Cornell University, Ithaca, USA; 3 Museo
4
Paleontologico Egidio Feruglio, Trelew, Argentina;
Denver Museum of Nature & Science, Denver, USA
We report ongoing investigations of the classic calderalake floras from Laguna del Hunco (LH) and Río
Pichileufú (RP) in Patagonia. 40Ar–39Ar analyses on sanidine from tuffs intercalated with the fossiliferous horizons give high-precision ages of 51.91 ± 0.22 Ma from LH (early Eocene) and 47.46 ± 0.05 Ma from RP
(middle Eocene). Stratigraphically controlled benchquarrying of >10,000 plant specimens from multiple horizons reveals hundreds of species at both sites, comprising a floral diversity among the highest known from the Eocene, and many important occurrences of fossil insects, insect-feeding damage, and vertebrates.
The biogeographic and paleoenvironmental signals from these assemblages have previously remained enigmatic due to undersampling, historical misidentifications, and lack of sustained taxonomic study. We find that the strongest affinity, now clear among conifers, angiosperms, and ferns, is to very humid, usually montane forests of tropical and subtropical Australasia.
All of the taxa listed below are present at LH, and most are also present at RP. The conifer assemblages include attached foliage and seed cone of Papuacedrus
(Cupressaceae); foliage, cone scales, and probable pollen cones of Agathis and Araucaria Sec. Eutacta
(Araucariaceae); and diverse Podocarpaceae, including foliage attached to seed and pollen cones of
Dacrycarpus , and foliage of Acmopyle , Retrophyllum , and Podocarpus . These conifer taxa today demonstrate physiological intolerance to drought (especially
Acmopyle and Dacrycarpus ) and are entirely confined to rainforest; nearly all only occur in Australasia. The fern floras display a similar, though more cosmopolitan pattern, and include fertile fronds of Todea
(Osmundaceae), Dicksonia (Dicksoniaceae), and
Sticherus (Gleicheniaceae). Among angiosperms, the
Australasian connection is strongly represented by fruits and male inflorescences attached to branches of
Gymnostoma (Casuarinaceae); compound leaves of
Akania (Akaniaceae); flower buds, infructescences, and leaves of Eucalyptus subgenus Symphyomyrtus
(Myrtaceae); fruits of Orites and diverse leaves of other
Proteaceae, and leaves and fruits of Atherospermataceae,
Monimiaceae, Lauraceae, and Cunoniaceae. The presence of abundant Eucalyptus subgenus
Symphyomyrtus in the ancient volcanic environment strongly suggests a vegetational mosaic of diverse rainforest alongside recent lava flows dominated by
Eucalyptus , exemplified by the modern analog E. deglupta (also subgenus Symphyomyrtus ) in calderas of
New Britain. Most of the taxa listed have Paleogene fossil records in southeastern Australia, and a few have been found in Antarctica; they document a long and complex history of retreat and survival in the face of severe climate change, regional extinction, and the movement of northern Australia into the tropics.
Neotropical affinities of the floras remain few and mostly montane, and affinities to Andean temperate rainforest are scarce. The extensive taxonomic similarity of ancient
Patagonian to modern Australasian, and to fossil southeast Australian, floras at the generic level is characteristic only during the Eocene. It is reduced in
Late Cretaceous, Paleocene, or younger Patagonian floras and appears to indicate extensive lineage mobility across warm high latitudes, followed by migrations into lower latitudes in response to cooling and drying. This scenario is analogous to well-documented distributional shifts of coeval Northern Hemisphere floras and faunas but was not previously observed on this scale in the Southern
Hemisphere.
Fossil and modern phylogenetic components of
Patagonian seed plant diversity
Nixon, KC 1 , Gandolfo, MA 1 , Hermsen, EJ 1 , Zamaloa,
1
MC 2
LH Bailey Hortorium, Dept of Plant Biology, Cornell
University, Ithaca, USA; 2 Departamento de Ciencia
Biológicas, Facultad de Ciencias Exactas y Naturales,
Universidad de Buenos Aires, Intendente Güira,
Argentina
Analysis of turnover in paleofloras suggest four distinct floristic intervals during the Tertiary of Patagonia in southern South America (Paleocene–Early Eocene,
Middle Eocene–Early Oligocene, Late Oligocene–Early
Miocene, and Middle–Late Miocene). These turnovers are significant and provide a basis for understanding the origin and evolution of modern Patagonian floras. We use the palynological and paleobotanical data summarized by Barreda and Palazzesi to analyze 33
Patagonian Tertiary formations (10 Paleocene–Early
Eocene sites, 9 Middle Eocene–Early Oligocene sites, 12
Late Oligocene–Early Miocene sites and 2 Middle–Late
Miocene sites). We augmented these data with recent
271

additions to the paleofloras of sites in Patagonia, including Salamanca, Laguna del Hunco, Rio Pichileufu,
Rio Nirihuau and Cullen Formations among others.
Modern floristic data from extant floras of Patagonia, northern Argentina, Australia, South Africa, New
Caledonia, New Zealand and selected Northern
Hemisphere sites including the arid west of North
America were then combined into a comprehensive matrix that was analyzed for clade correspondence using a 'supertree' generated from the most recent molecular analyses for all clades represented in the fossil floras of
Patagonia. A comparison of results using different phylogenetically-based approaches will be presented.
This provides both an understanding of the biogeographic affinities of the southern cone Tertiary paleofloras as well as the ability to generate phylogenetic indices for shared clades among fossil and modern floras.
Some interesting elements of the Tertiary flora of
India
Mehrotra, R 1
1 Birbal Sahni Institute of Palaeobotany, Lucknow, India
As per the existing palaeocontinental maps India started its northward journey during the Aptian as an isolated island continent after being separated from the other
Gondwanaland continents. It collided with the Asian landmass during the Eocene resulting in the formation of land connections at the end of the Oligocene/Early
Miocene. Due to this long isolation it is expected that
India should have an endemic flora, which is not the case. This is why the study of its palaeovegetation becomes so interesting. The Palaeogene flora shows the presence of some elements of the Gondwanaland continents, while the Neogene flora is dominated by the
Southeast Asian and Chinese elements. The most important amongst them is the Dipterocarpaceae, a typical Southeast Asian family, which appeared in India only during the Middle Miocene. The present day flora seems to be an admixture of various floras migrated through various routes at different time. The entry of angiosperms might have been late in India (not before
Maestrichtian) in comparison to their appearance on other continents. Palms which were dominant during the
Maestrichtian–Danian declined afterwards up to
Oligocene and were rare during the Neogene.
Gymnosperms are represented by the Podocarpaceae and
Araucariaceae during most of the Tertiary. The former is at the verge of its extinction, while the latter became extinct from India. Several new families (Pinaceae,
Cephalotaxaceae and Taxodiaceae) started inhabiting the
Himalayan region at the end of the Pliocene. The Tertiary flora of India was dominated by tropical evergreen to semi-evergreen and littoral and swampy forests having tropical warm and humid climate. The vicinity of the
Tethys Sea during the Palaeogene could be attributed for the same. Middle Miocene was the time of maximum diversification and migration of taxa. The Himalayan uplift caused further changes in the physiography which in turn affected the climate. As a result cold and dry conditions prevailed in the Himalayan foot-hills due to which subtropical and temperate elements came into existence from the nearby Tibettan region where they were growing luxuriantly. The Karewa flora of the Late
Plio-Pleistocene is the best example to prove it.
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Mid-Cenozoic paleofloras of New Zealand: reconstructing the vegetation complexity of an isolated landmass in the Southern Hemisphere
1
Lee, DE 1 , Bannister, JM 1 , Conran, JG 2 , Mildenhall, DC 3
Dept of Geology, University of Otago, Dunedin, New
Zealand; 2 Australian Centre for Evolutionary Biology and Biodiversity, School of Earth and Environmental
Sciences, University of Adelaide, Australia; 3 GNS
Science, Lower Hutt, New Zealand
The fossil record of the New Zealand Cenozoic flora is perhaps the most continuous in the Southern
Hemisphere, and provides an exceptional account of the changing vegetation, environments and climate on an oceanic archipelago. The comprehensive pollen and spore record is now being supplemented by integration of new data from a diverse range of macrofossils occupying a broad array of habitats, including several sites such as maar lakes that were effectively closed systems. Modern techniques employed include investigation of cuticular anatomy of mummified leaf compressions of conifers, monocots and dicots, anatomy of leaf phytoliths, 3-D analysis of the structure of silicified and lignified wood, chemical analysis of resin, associations between flowers and in situ pollen, ferns and in situ spores, and the incorporation of results into new molecular phylogenies.
Several examples will be discussed. The first is a late
Eocene flora from an alluvial flood plain deposit that includes wood from in situ trees, bark, fertile fern fronds, leaves with preserved cuticle, and in situ epiphyllous fungi. The second is a late Oligocene leaf bed in lignite that includes conifer foliage, wood and resin, as well as monocot and dicot leaves, and which represents woody vegetation growing in and around a nutrient-poor ombrotrophic mire. The third, preserved in an early
Miocene silcrete deposit, includes silicified wood, 3-D fruits and seeds from forest trees and shrubs that grew on an alluvial or deltaic flood plain. The final examples are two early Miocene maar lake deposits in which diatoms and algae are present, together with flowers containing in situ pollen, mummified leaves with exquisitely preserved cuticle, and fruits from the vegetation that surrounded the small lakes. Plant macrofossils represented at these sites include species of Araucaria , Agathis , Dacrycarpus ,
Dacrydium , Phyllocladus , Podocarpus , Prumnopitys ,
Banksia , Elaeocarpus , Fuchsia , Gymnostoma ,
Mallotus / Macaranga , Myrsine , Nothofagus ,
Weinmannia , Araliaceae, Myrtaceae and Sapindaceae together with a wide variety of monocots typical of present-day New Zealand vegetation such as Astelia ,
Cordyline , Luzuriaga , orchids, Phormium , Ripogonum and Typha . The implications of these records for a better understanding of Southern Hemisphere plant biogeography will be discussed.

Evolutionary transitions from outcrossing to selfing in plants: insights from molecular population genetics and genomics
Barrett, S 1 , Ness, R 1 , Siol, M 1
1 University of Toronto, Toronto, Canada
The origin of predominant self-fertilization (autogamy) from obligate cross-fertilization is considered the most frequent transition in flowering plant reproductive systems. Heterostylous groups provide a rich source of reproductive diversity for investigating this transition.
This is because in many taxa obligate outcrossing, enforced by heteromorphic self-incompatibility, has been replaced by predominant selfing as a result of the origin and spread of homostylous variants, with the capacity for autonomous self-pollination. Population genetic theory predicts that increased homozygosity due to inbreeding reduces the effective size of populations and increases the extent of linkage disequilibrium in highly selfing populations. Combined with demographic factors associated with genetic bottlenecks and subdivision, this can lead to a reduction in diversity within populations and increased differentiation among populations. In addition, slightly deleterious or advantageous mutations can be rendered effectively neutral in selfing populations because of the increased importance of genetic drift.
These processes are predicted to reduce selective constraints on protein sequences and cause relaxed selection for biased codon usage. Here, we investigate these theoretical predictions in the neotropical annual plant Eichhornia paniculata (Pontederiaceae) by examining the patterns and distribution of nucleotide variation in protein-coding genes across the genome.
This species occupies ephemeral ponds, ditches and rice fields and occurs mainly in N.E. Brazil, with smaller concentrations of populations on the Caribbean islands of
Jamaica and Cuba, and isolated populations in Nicaragua and Mexico. We sequenced 10 EST-derived nuclear loci from 25 populations representing much of this geographic range. We found that inbreeding reduced variation within populations and was associated with greater differentiation among populations, as predicted.
However, significantly reduced diversity was only evident in highly selfing populations and outcrossing populations and those composed of a mixture of selfing and outcrossing plants maintained similar levels of diversity. Population size interacted strongly with mating system and explained more of the observed variation in diversity within populations; small populations were largely selfing and large populations largely outcrossing.
Coalescent simulations provided evidence of a moderate bottleneck associated with colonization of the Caribbean from Brazil ~125,000 years before present. Although our results in E. paniculata confirm the predicted reduction in diversity associated with the evolution of selfing, the amounts of diversity were higher than expected under long-term equilibrium. This is probably due to the recent multiple origins of selfing from diverse outcrossing populations. To investigate the effect of inbreeding on the accumulation of deleterious mutations in proteincoding genes we used Illumina short-read sequencing to assemble, de novo, the floral transcriptomes of an outcrossing and three independently derived selfing lineages of Eichhornia . We identified ~8000 orthologous sequences totalling ~3.5 Mb of coding DNA. Several tests of selection provided results that were consistent with the occurrence of purifying selection across the transcriptome in selfing lineages, especially with respect to weakly selected synonymous changes that affect codon usage. The occurrence of multiple independent origins of self-fertilization among populations of E. paniculata provides outstanding opportunities for investigating the molecular population genetic and genomic consequences of selfing in plants.
The evolutionary significance of geographic variation in the mating system within plant species
Eckert, C 1 , Dart, S 1 , Lopez Villalobos, A 1
1 Queen's University, Ontario, Canada
The relative importance of outcrossing versus selffertilization is a major component of mating system variation in flowering plants. Divergence in the mating system often occurs between closely related species suggesting that floral traits affecting mating are evolutionarily labile and that transitions in the mating system are very frequent. However, empirical demonstrations of the selective factors responsible for these transitions are few. Species exhibiting marked mating system variation among populations might provide clearer insight into the evolutionary processes involved. Mating system variation among closely situated populations suggest that evolutionary divergence is relatively recent, and substantial gene flow between divergent populations further suggests that the selective factors responsible are still operating. The geographic patterning of intraspecific mating system variation may provide clues as to the selective agents involved. We are investigating mating system differentiation within
Camissoniopsis cheiranthifolia (Onagraceae), a shortlived, insect-pollinated, herbaceous plant endemic to the
Pacific coastal dunes of North America. Plants in San
Diego County (California) produce large, rewarding, self-incompatible flowers. Further north to Point
Conception, flowers are large but fully self-compatible.
North from Point Conception to southern Oregon, south into Baja California and on the Channel Islands plants produce small, self-pollinating and relatively short-lived flowers, suggesting more than one independent transition to selfing. Historical surveys of insect flower visitors suggest that a transition to selfing happens at Point
Conception because the coast north of the point experiences stronger winds and more frequent fog, which compromises pollinator visitation. Hence selfing is hypothesized to be a mechanism of reproductive assurance, a frequently invoked but rarely tested advantage of selfing. Consistent with this hypothesis, fruit set in large-flowered populations is much lower and more variable than in small-flowered populations.
However, pollen supplementation experiments indicate that, although outcross pollen limitation is somewhat weaker in small-flowered populations, it is mild in largeflowered populations and does not account for geographical variation in fruit set. Rather, substantial fruit losses in large-flowered populations are caused by a
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microlepidopteran ( Mompha sp.), the larvae of which damage developing flowers causing abortion before fruit maturation. Although this parasite is much less frequent in small-flowered populations, it does not preferentially parasitize larger flowers and probably does not exert selection favouring reduced flower size and selfing.
Estimation of the mating system using allozyme markers indicates that although large-flowered populations tend to outcross more than small-flowered populations, the mating system varies somewhat independently of flower size, even in self-incompatible populations.
Unexpectedly, populations that produce very small flowers often exhibit substantial outcrossing (20–60%) despite inbreeding depression being very weak (< 0.1) in these populations. Moreover, the flowers of predominantly selfing C. cheiranthifolia populations are much larger than those of closely related, strongly selfing species. Only in a single northern California population of C. cheiranthifolia has the transition to self-fertilization been completed with the development of obligate cleistogamy. Taken together, our results from C. cheiranthifolia do not support a simple interpretation involving selection for reproductive assurance causing mating system diversification.
Raindrops drive the evolution of floral form
Huang, S-Q 1
1 College of Life Sciences, Wuhan University, China
Flowers exhibit adaptive responses to mutualistic pollinators and antagonistic herbivores as well as the physical environments. Early researchers, including
Charles Darwin, have speculated that flowers may have evolved certain traits or structures to protect themselves against the damaging effects of rain. Rain can drop off the whole flower, wash away pollen grains, reduce pollen viability and dilute nectar, but the selective role of rain on the evolution of floral traits has been rarely experimentally tested. We first investigated flower performance in rain to examine whether flower structures protect pollen from rain contact in 80 flowering species from 46 families with diverse floral shapes and pollination modes. We then compared pollen longevity in dry, pure water and solutions with different sucrose concentrations to test pollen resistance to water. Pollen viability test showed that pollen longevity in all studied species was greatly shortened by wetting. We found that pollen of species with complete protection by flower structures was susceptible to water damage and a high proportion of resistant pollen occurred in unprotected species. Our observations showed that variation in pollen performance in water was associated with different floral forms, in support of the functional hypothesis that flower structures protect susceptible pollen from rain. We further investigated flower performance in numerous species in an alpine community to examine the effect of rain on the evolution of floral form. The main results will be presented at the IBC2011 conference.
Intraspecific variation in pollinators as a driver of floral diversification within the sexually deceptive daisy
Gorteria diffusa
De Jager, ML 1 , Ellis, AG 1
274
1 Stellenbosch University, Matieland, South Africa
Of the various ways that flowering plants ensure pollination sexual deception is arguably the most spectacular. Deceptive flowers entice male pollinators by mimicking the mating signals of female insects.
Previously thought to be exclusive to the Orchidaceae, this mode of pollination was recently discovered in the
South African daisy, Gorteria diffusa . This species comprises 14 geographically distinct floral morphotypes, all predominantly pollinated by the beefly Megapalpus capensis . Three of the floral morphs employ complex petal spots to elicit mating behaviour from male flies, whilst the other morphs only induce inspection or feeding behaviour. Although intraspecific variation in pollinators has been considered a potential driver of floral divergence, few studies have experimentally investigated this mechanism. Here we explore the potential role of sex-dependant preferences of male and female flies as well as geographic variation in the preferences of male flies in the diversification of mimetic flower ornaments within this species. We designed a series of experiments to investigate the influence of visual, tactile and olfactory petal spot components on beefly visitation. Here we show that males and females have contrasting preferences with males generally preferring all components of petal spots, including UV reflective highlights. Females generally exhibited random preference for these components, but significantly avoided UV reflective highlights. This suggests that specializing on the attraction of mate-seeking males through the production of UV reflective highlights might result in reduced visitation by food seeking females.
Interestingly, males did not exhibit preference for floral scent. Since olfaction is the dominant floral cue employed by orchids to deceive their male pollinators this implies that sexual deception in daisies might have occurred through different evolutionary pathways to those in orchids. To test for floral divergence through geographically structured variation in male fly preferences, we used a reciprocal test whereby we presented two distinct sexually deceptive floral morphs to the two evolutionary lineages of male flies they are associated with. We found no evidence for local adaptation on the basis of male fly preference, but instead males exhibited a consistent hierarchy of preference for sexually deceptive morphotypes across their range.
Experienced male flies also exhibited a significant reduction in mating intensity relative to naïve males which had never encountered G. diffusa . This learned discrimination of petal spots as female mimics mirrors responses of male hymenoptera to sexually deceptive orchids and was confirmed in learning experiments.
These results suggest that divergent selection imposed by male and female flies may have contributed to the incredible floral variation within G. diffusa and propose the exciting possibility that spot elaboration in sexually deceptive morphs might arise through a co-evolutionary arms race between plant and pollinator.
The reproductive biology of two Himalayan alpine gingers (
Roscoea spp., Zingiberaceae) in China: pollination syndrome and compensatory floral mechanisms
Zhang, Z-Q 1 , Kress, WJ 2 , Li, Q-J 3

1 Key Laboratory of Biodiversity and Biogeography,
Kunming Institute of Botany, Chinese Academy of
Sciences; 2 National Museum of Natural History,
Smithsonian Institution, Washington, DC, USA; 3 Key
Laboratory of Tropical Forest Ecology, Xishuangbanna
Tropical Botanical Garden, CAS, China
According to the concept of pollination syndromes, floral traits reflect specialization to a particular pollinator or set of pollinators. However, the reproductive biology of endemic, and often specialized, plants may require increased attention as climate change accelerates worldwide. Species of Roscoea , endemic to the
Himalayan region, have striking orchid-like flowers with long corolla tubes, suggesting pollination by longtongued insects. Until now the reproductive biology of species of Roscoea has been poorly documented. We investigated the floral biology, breeding system, and pollination ecology of R. cautleoides and R. humeana , from Hengduan Mountains, a global biodiversity hotspot in SW China. We also tested whether the floral longevity helps to increase pollination success. Pollination experiments showed that the two species were selfcompatible and depended on insects for fruit production.
Over several flowering seasons we did not observe any suspected pollinators with long tongues, which would match the corolla tube, to visit the flowers in the centers of distribution. The principal pollinators observed were pollen-collecting generalist bees with low visitation frequencies. In general members of the ginger family are characterized by short-lived, usually one-day flowers, but the floral longevity of R. cautleoides and R. humeana is six and eight days, respectively. Removing stigmas decreased fruit set in both study populations. Our results suggest that the original pollinators may have been longtongued insects which are now absent in the Chinese
Himalayas as habitats respond to climate change.
However, the long-lived and self-compatible flowers coupled with the presence of generalist pollinators are traits that have allowed these gingers to reproduce and continue to persist in alpine habitats.
Why package pollen in groups?
, Moat, J 1 , Lewis, G 1
1
Banks, H 1
Royal Botanic Gardens Kew, UK
Pollen dispersal and pollination syndromes are of fundamental importance to the evolution of flowering plants, and pollen has evolved into a wide range of forms in response to a plethora of ecological pressures, with those forms constrained by inheritance, physical and physiological requirements, and developmental processes. From the pollen record it is difficult to recognise pollen types that directly indicate habitat.
Using GIS data, species distributions taken from herbarium specimens, and pollen morphological research, we present a study that connects ecological habitat with pollen structure. Most angiosperms release their pollen grains as individual units (monads), but aggregations of coherent groups of pollen units (polyads) have evolved a number of times. Both polyads and monads occur in early diverging taxa of mimosoid legumes (Leguminosae, subfamily Mimosoideae). Some mimosoid polyads have evolved as part of a specialised reproductive syndrome; the number of pollen units in the polyad matches the number of ovules in the ovary, and the polyad exactly fits the size of the stigmatic opening.
Other studies have shown that pollen aperture number is positively correlated with pollen germination speed.
Multiple apertures ensure that a pollen grain has contact with stigma fluids allowing fast re-hydration irrespective of the orientation it is deposited in. However, pollen with few apertures has a better survival rate. The more gaps
(apertures) that there are in the pollen wall, the greater the speed of dehydration plus increased possibility of pathogenic attack. In mimosoids, individual pollen units within the polyads have relatively small apertures, maintaining viability, but many apertures per polyad
(through which fluids are interchangeable; polyads function as a single harmomegathic unit), providing efficient germination. We found that in closely related endemic Malagassy mimosoid legume genera that are known to have some species with monads and some with polyads, species that release pollen in polyads are distributed in drier habitats than those that release their pollen as individual grains.
Gametophytic competition as an ecological process
Harder, L 1 , Aizen, M 2
1 University of Calgary, Alberta, Canada; 2 INIBIOMA-
CONICET, Argentina
Seed production depends on the production of ovules, their fertilization, and subsequent embryo survival and development. Because of the serial dependence of these processes, limited performance during early stages necessarily diminishes options for later stages and poor performance during a late stage can restrict ultimate success, even though early stages functioned at capacity.
Nevertheless, most ecological and evolutionary studies of plant reproduction focus on the early stage of plantpollinator interaction and/or the final stage of seed production. In contrast, less than 10% of studies of the influence of pollination of reproductive performance consider the intervening stages of pollen deposition on stigmas, pollen-tube growth and ovule fertilization, so that post-pollination (progamic) processes are typically either ignored or treated as a 'black box'. Although pollen germination and pollen-tube growth fundamentally involve biochemical and physiological processes, an ecological analogy may provide insights into their general consequences for siring success and seed production. Specifically, we consider progamic success as the outcome of competition governed by the availability of 'resources', including nutrients and/or space, and the density and activity of competitors. As independent haploid organisms, male gametophytes of angiosperms complete their lives in localized populations in the complex and dynamic environment provided by the pistils of conspecific sporophytes. Styles represent a competitive arena for such populations by providing resources that fuel pollen-tube growth. Therefore, if stylar resources are limiting, male gametophytes can compete in manners that depend on their characteristics and those of the resource supply (i.e., the pistil).
'Contest', or exploitative, competition could arise if pollen grains differ extensively in the timing of their arrival on a stigma and/or germination time, or in tube growth rate, whereas 'scramble', or interference, competition could occur when germination time differs
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little and pollen tubes grow at equal rates, so they compete as an advancing front. According to this ecological analogy, the nature and magnitude of resource competition in the style should influence the number and quality of the seeds set in individual fruits. Observed differences among species in the relation of the number of pollen-tubes entering ovaries or seed production to pollen receipt by stigmas support the contrasting expectations of contest and scramble competition.
Furthermore, consideration of the implications of progamic resource competition reveals explanations for enigmatic aspects of plant reproduction and enriches understanding of the incidence and nature of sexual selection in plant populations.
Evolution of the mating system and sex allocation in plant metapopulations
Pannell, J 1
1 University of Oxford, UK
The optimal investment of resources towards the male versus female functions of plants is thought to depend on both the shape of the fitness gain curve for the population and the relatedness between mating parents at loci with an influence on allocation. Most models of sex-allocation evolution, and sexual-system evolution more generally, formulate their predictions in terms of gain-curve and relatedness parameters (as described, for example, by inbreeding coefficients) that are implicitly fixed for the context under focus. But how should sexual-system evolution be affected by a species’ range expansion, by ongoing colonization dynamics in a metapopulation, or by small-scale patch dynamics within populations? In each of these scenarios, both the shape of fitness gain curves and the relatedness between parents may fluctuate widely, so that equilibrium-based models are not likely to apply. Here, I formulate theory for the evolution of the sex allocation in terms of the well-known hierarchical inbreeding coefficients that suggests that, in plant metapopulations, F_st may often be a better predictor of the sex allocation than F_is. I illustrate a test of the theory using comparisons of hermaphroditic sex allocation among metapopulations of the plant
Mercurialis annua that are characterized by contrasting sexual systems. In M. annua , patterns of sex allocation appear to have been influenced by colonization during range expansions, within regions as part of ongoing metapopulation dynamics, as well as through the colonization of patches at small spatial scales within extended local populations. Finally, I discuss the importance of the genetic architecture of sexual-system traits, rather than just their phenotypes, for predicting the ratios of different allocation and mating strategies subject to frequency-dependent selection.
Mechanisms governing sex ratio variation in dioecious plants: the role of demographic and genetic factors in
Rumex hastatulus
Field, DL
Canada
1 , Pickup, M 1 , Barrett, SCH 1
1 Dept of Ecology and Evolution, University of Toronto,
Although the majority of plant species are hermaphrodites, dioecious species (separate male and
276 female plants) represent an important form of reproductive diversity in angiosperms. Despite a strong theoretical basis, there is currently very little known about the mechanisms governing sex ratio variation in plants. In a survey of 126 dioecious species we found nearly two thirds of the species exhibited biased sex ratios, with male bias twice as frequent as female bias.
The frequency of departures from the theoretical expectation of 1:1 suggests that several mechanisms
(both pre- and/or post-zygotic) may contribute to biased sex ratios. In addition, there is some evidence that species with sex chromosomes are more likely to exhibit female biased sex ratios. Here, we investigate the annual, wind-pollinated species Rumex hastatulus to determine to what extent populations show biased sex ratios and if so what genetic and ecological mechanisms may be responsible. Rumex hastatulus represents a valuable system to examine these questions because of its annual life history and polymorphic sex determining system, with two disjunct heterogametic sex chromosome races
(Texas race: Females XX, Males XY, and North Carolina race: Females XX, Males XY1Y2). A particular focus of our study was to assess the extent to which the local mating environment may contribute to biased progeny sex ratios by influencing pollen capture and the degree of gametophytic competition. A survey of 46 populations of
R. hastatulus (23 from each chromosome race) revealed consistently female-biased sex ratios (mean sex ratio =
0.62, SE = 0.01). A relation between male proximity and the sex ratios of offspring from females was found, based on progeny testing open-pollinated families in six of these populations. Females located in close proximity to males produced more strongly female biased sex ratios compared to more isolated females. These results suggest that male proximity may influence progeny sex ratios by affecting pollen loads and the strength of gametophytic competition. To examine the influence of pollen loads on fertilization success (pollen limitation) and progeny sex ratios, we experimentally added specific numbers of pollen grains ranging from 1 to > 100 to female stigmas.
Pollen load significantly predicted fertilization success, with consistently high seed set (> 80 %) when > 10 pollen grains were applied. We predicted an association between pollen load and the probability of producing female progeny due to greater pollen competition between male and female determining microgametophytes when pollen loads are high. We further examined this prediction using field based experimental arrays in which we manipulated both frequency (sex ratio) and plant density (plants m-2) in a factorial design combining three frequency (male bias, equality, female bias) and three density (high, medium, low) treatments. We assessed the interaction between frequency and density by measuring pollen loads, seed set and progeny sex ratio for focal females within each array. Collectively, the results of our experiments provide novel insights into the influence of demographic factors on the mechanisms governing sex ratios in dioecious plants.
Phylogeography of
Lycium californicum
(Solanaceae) and intraspecific variation in sexual system, ploidy level, and floral morphology
Miller, JS 1 , Levin, RA 1
1 Amherst College, Amherst, MA, USA

Lycium (Solanaceae) is a genus of ca. 90 species of perennial shrubs occurring in arid to semi-arid habitats worldwide. Members of Lycium vary in sexual function; whereas most species are cosexual and produce hermaphroditic flowers, others have gender dimorphism ranging from separate female and hermaphroditic plants to complete dioecy. There are ten named species with gender dimorphism, and dimorphism has arisen convergently in North America and southern Africa, having evolved multiple times independently within each region. The evolution of gender dimorphism in Lycium is especially interesting because it has occurred on a phylogenetic background of self-incompatibility. It has been proposed that gender dimorphism evolves in
Lycium following polyploidy, which acts as a trigger for this transition because it disrupts the incompatibility system. Previous studies have confirmed that gender dimorphic species are polyploid, and cosexual species diploid. One exceptional North American species,
Lycium californicum , shows intraspecific polymorphism in sexual strategy and initial surveys of this species indicate that populations are either cosexual or gender dimorphic. Here, we expand sampling of Lycium californicum to include individuals from populations across the entire species range, which extends north from the Channel Islands of California south to the tip of Baja
California Sur, and east to mainland Mexico and
Arizona. We use a combination of nuclear and plastid data to explore the phylogeographic history of this taxon and, specifically, the relationship of cosexual and dimorphic populations. We also investigate the floral morphology of mating types within and across populations and at a geographic scale. Finally, we assess the association of ploidy level and sexual strategy across this larger set of thirty populations.
Breeding system evolution in heterostylous– homostylous
Primula
, with special reference to section
Aleuritia
, Keller, B 1 , Kelso, S 2 , Conti, E 1
1
De Vos, J 1
Institute for Systematic Botany, Universtiy of Zurich,
Switzerland; 2 Dept of Biology, Colorado College,
Colorado Springs, USA
Flowering plants are remarkably diverse in floral systems and reproductive strategies. Heterostyly, which sparkled the interest of evolutionary biologists in reproductive systems for over 140 years, is a polymorphic breeding system that enhances outcrossing. The two (distyly) or three (tristyly) floral morphs of heterostylous species are characterized by reciprocal placement of anthers and stigmas and (usually) self- and intra-morph incompatibility. The system has evolved presumably independently in more than 20 families of flowering plants. Homostylous, self-compatible species have often evolved within groups that are predominantly heterostylous, providing valuable model systems to study evolutionary transitions in reproductive strategies. The fact that homostylous species occur more frequently in
Arctic and alpine environments, where pollination services are less reliable than in more temperate environments, has often been interpreted as evidence for the hypothesis that homostyly provides an adaptation for reproductive assurance. However, different strategies may allow plants to reproduce in habitats with reduced pollinator services. Besides autogamy, plants can also invest more in floral display for pollinator attraction, or increase the number of ovules per flower to optimize seed production when pollination events are scarce. Our model system Primula section Aleuritia contains homostylous and heterostylous species, from lowland,
Arctic and alpine habitats, of a wide range of ploidy levels, and therefore provides an excellent model system to study evolutionary transitions in breeding systems. In our research, we applied three different approaches. With pollinator exclusion and emasculation experiments in the field, we tested whether homostyly provides reproductive assurance in the alpine Primula halleri . In these experiments, we quantified the effect of minute quantitative variation in the relative position of anthers and stigma within a flower on reproductive success.
Subsequently, at the sectional level, we assessed the contribution of variation in floral morphology, fecundity
(especially pollen/ovule ratios), and breeding system to different adaptive reproductive strategies, to elucidate how homostylous species have adapted to habitats with reduced pollinator fauna. Finally, at the family level, we developed a robust phylogenetic framework that allows us to statistically test hypotheses of correlations between breeding systems, floral morphology and diversification rates, using Bayesian approaches.
Does the likelihood of an Allee effect in plant reproductive success depend on the type of pollinator?
Duffy, K 1 , Johnson, S 1
1 School of Biological and Conservation Sciences,
University of KwaZulu-Natal, South Africa
Allee effects (positive relationships between numbers of conspecific individuals on individual fitness) can be important in governing the reproductive success of animal-pollinated, self-incompatible plants. This is because larger floral displays are attractive to animal pollinators as they maximise their foraging efficiency.
However, once attracted to populations, local pollinator taxa can differ in the efficiency with which they pollinate flowers; hence may differ in whether they cause an Allee effect in plant reproduction. We examined the reproductive biology of the self-incompatible Kniphofia linearifolia (Baker), which has two types of pollinator; birds (mainly sunbirds and weavers) and native bees
(honeybees and solitary bees). We made replicated pollinator observations in populations of varying size, density, and isolation. To test for differences between the two pollinator types, we caged K. linearifolia individuals in each population to exclude bird visitors. We found a strong Allee effect related to bird visitation, with increases in population size increasing both bird visitation and overall reproductive success. No Allee effects were related to bee visitation. Although bees did contribute to fruit and seed set, caged plants had lower fruit and seed set overall, indicating that birds are more efficient pollinators. This study demonstrates that the likelihood of an Allee effect in plant reproduction can depend on the type of pollinator available in a population. From a conservation perspective, small populations (<100 flowering individuals) of K. linearifolia may not be able to attract enough bird pollinators. Such populations may be reliant on
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inefficient bee pollinators and vulnerable to inadequate levels of reproduction, which will negatively affect their long-term survival.
Expanding the Bayesian Island Biogeography Model to continental scenarios: the Rand Flora pattern
Sanmartin, I 1 , Anderson, CL 2 , Aldasoro, JJ 1 , Ronquist,
1
F 3
Real Jardin Botanico, CSIC, Spain; 2 Dept of Plant and
3
Environmental Sciences, Göteborg University, Sweden;
Dept of Entomology, Swedish Museum of Natural
History, Sweden
Markov chain Monte Carlo (MCMC) Bayesian inference techniques have become very popular in phylogenetic inference because of the relative ease with which these techniques allow us to infer evolution using more complex and realistic models. However, it has taken much longer time to transfer Bayesian techniques, and in general statistical inference methods, to the field of biogeography, where parsimony has been the favourite mean of inferring the past. This reliance on parsimony has made it difficult to incorporate relevant evidence other than the tree topology and species distributions to biogeographic analyses, for example: the times of divergence between lineages or information on the connectivity of geographical areas through time. Here I present a Bayesian statistical approach that integrates phylogenetic and biogeographic uncertainty to estimate carrying capacities (equilibrium frequencies of species diversity) and rates of dispersal/ biotic migration between geographically isolated areas, using DNA sequence data and species distributions. An advantage of the method is that biogeographic parameters can be estimated across groups differing in their age, evolutionary rate, and/or dispersal capabilities, so it can be used for inferring significant patterns or trends at regional level. So far, the
BIB model has been used in an island context (i.e., areas separated by oceanic barriers). Here, we show that the
BIB model may be useful in a continental setting where areas are separated by ecological barriers, and where the number of inferred dispersal/migration events between areas is low. We explore its use for disentangling the evolutionary origins of a continental-scale floristic pattern (the ‘Rand Flora’) that evolutionarily relates disjunct floras along the margins of the African continent.
An evaluation of new parsimony-based versus parametric inference methods in biogeography: a case study using the worldwide plant family
Sapindaceae
Buerki, S 1 , Forest, F 1 , Sanmartin, I 2
1 Molecular Systematics section, Jodrell Laboratory,
Royal Botanic Gardens, Kew, UK; 2 Real Jardin
Botanico, CSIC, Madrid, Spain
Recently developed parametric methods in historical biogeography allow researchers to integrate temporal and palaeogeographic information into the reconstruction of biogeographical scenarios, thus overcoming a known bias of parsimony-based approaches. Here, a parametric method, dispersal-extinction-cladogenesis (DEC), is compared against a parsimony-based method, dispersalvicariance analysis (DIVA), which does not incorporate branch lengths but accounts for phylogenetic uncertainty through a Bayesian empirical approach (Bayes–DIVA).
The benefits and limitations of each method are analyzed using the cosmopolitan plant-family Sapindaceae as a case-study. Phylogenetic relationships and divergence time estimation were estimated by Bayesian inference on a large dataset representing generic diversity within
Sapindaceae. To account for phylogenetic and dating uncertainties in biogeographical reconstructions the analyses were done over a sample of trees from the posterior distribution of the Bayesian inference. We compared biogeographical scenarios between Bayes–
DIVA and two different DEC models: one with no geological constraints; and another that employed a stratified palaeogeographic model in which dispersal rates were scaled according to area connectivity across four time slices, reflecting the changing continental configuration during the last 110 Ma. Despite differences in the underlying biogeographical model, Bayes–DIVA and DEC inferred similar biogeographical scenarios. The main difference was in the timing of dispersal events – which in Bayes–DIVA sometimes conflicts with palaeogeographic information – and the higher frequency of terminal dispersal events compared with DEC.
Uncertainty in divergence time estimations influenced both the inference of ancestral ranges and the decisiveness with which an area can be assigned to a node. By considering lineage divergence times, the DEC method gives more accurate reconstructions in agreement with palaeogeographic evidence. In contrast, Bayes–
DIVA showed the highest decisiveness in unequivocally reconstructing ancestral ranges, probably related to its ability to integrate phylogenetic uncertainty. Care should be taken in defining the palaeogeographic model in DEC because of the possibility of overestimating the frequency of extinction events, or of inferring ancestral ranges that are outside the extant species ranges, owing to dispersal constraints enforced by the model. The widespanning spatial and temporal model proposed here could prove useful for testing large-scale biogeographical patterns in plants.
The Bayes–DIVA method and its application for testing biogeographic origins of inter-continental disjunct endemics
Harris, AJ 1 , Xiang, J 2
2
1 Oklahoma State University, Dept of Botany, USA;
North Carolina State University, Dept of Plant Biology,
USA
The use of phylogenetic analysis to evaluate hypotheses of historical biogeography has been faced with challenges that include uncertainties in resolution in topology and in ancestral range reconstruction. This may be overcome with our recent modification to the Bayes–
DIVA method, which iterates a DIVA analysis over a sample of Bayesian trees and then averages the support
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for a geographic range at a node over all trees to estimate the probability for each possible range at the node. This modified Bayes–DIVA method estimates support for geographic ranges reconstructed for the stem node of a crown group which has an unresolved sister group, and thus is particularly useful for strongly supported crown groups for which relationships among them remain unresolved – a common problem that arises from insufficient data or rapid radiation. A previous limitation to this approach was the absence of an automated method for performing analyses on large or many datasets, which is now possible using the software S-DIVA. Here we examine the ancestral ranges of endemic clades of disjunct arcto-tertiary remnant genera to test the alternative hypotheses that these lineages represent isolated descendents of more widely spread ancestors vs. having evolved in situ. We are analyzing published sequence data of approximately 30 genera using the
Bayes–DIVA approach implemented in S-DIVA.
Preliminary results indicate that the Asian endemics appear to have diversified in situ more frequently than lineages endemic to western North America, eastern
North America, Europe, and Central America. Presence of endemic lineages in the latter areas is better explained by vicariance than in situ diversification. Thus, non-
Asian endemics more often appear to be of relictual origin. These results are consistent with previous studies showing that more speciation of arcto-tertiary lineages has occurred within Asia when compared to other
Northern Hemisphere continental areas. This methodology may also be applicable at finer scales for the identification of the geographic origins of endemic species.
A temporal framework for studies in molecular dating and historical biogeography of angiosperms
Anderson, CL 1, 2 , Buerki, S 3 , Sanmartín, I 1
1 Real Jardin Botanico, CSIC, Madrid, Spain;
Plant and Environmental Sciences, Göteborg University,
3
2 Dept of
Sweden; Molecular Systematics Section, Jodrell
Laboratory, Royal Botanic Gardens, Kew, UK
Reconstructing the spatio-temporal evolution of organisms has become a major theme in evolutionary biology. Estimates of lineage divergence times as reconstructed from molecular dating or the fossil record are commonly used in biogeographic reconstructions, either as a reference for interpreting analyses or, more recently, as input data in model-based biogeographic analyses. Several large-scale studies attempting to date major divergences within the angiosperms, or within some of the main lineages (e.g. monocots, basal eudicots, asterids), have been published recently. Even though these studies are often taxonomically well sampled, only the oldest fossil occurrence for each major lineage has been used as potential calibration point to date the deepest divergences. This has in many cases resulted in severely underestimated crown group ages for nodes closer to the leaves, e.g. families, as can be observed when comparing molecular estimates with the known fossil record. Moreover, age estimates from these studies are often used as secondary calibration points for molecular dating of plant lineages of 'lower taxonomical level' with scarce or no fossil record. A common approach is to sample a larger group than the group of interest and do a 'global dating' analysis based on a secondary calibration point, and then use ages from this study as a tertiary calibration point. The errors introduced by this approach can be severe, and due to different sampling and dating methods, partly overlapping phylogenies that have been dated independently in different studies may show highly divergent results.
Here, we propose a new approach to reduce the potential errors stemming from use of studies with a focus on deeper divergences, and to specifically improve age estimates for angiosperm families. This is done by greatly improving two important factors: incorporating as much fossil evidence as possible, and substantially increasing taxon sampling. Beside a set of generally accepted fossils of major lineages, hundreds of fossils of lower taxonomic rank have been included. The coding plastid rbcL region is the best candidate to achieve this task since it is widely used to infer relationships within angiosperms and therefore represents most of the diversity of this group, which is important to avoid, as far as possible, unbalanced taxon sampling. We compiled a large rbc L dataset (~3200 sequences), covering all major angiosperm lineages, including the vast majority of families. Phylogenetic relationships were constrained to be congruent with APGIII. Lineage divergence times were estimated using penalized likelihood, since this method has been proven relatively stable, whilst able to deal with large datasets. Uncertainties in topology and branch lengths were accounted for through the use of a
MCMC Bayesian sampling approach. Although the resulting age estimates are still open to refinement, they provide us with a stable framework that is useful for secondary calibrations and placeholders when inferring the spatio-temporal evolution of less inclusive taxonomic groups. This framework is especially important when used consistently in studies comparing the evolutionary history of different groups, such as in the Rand Flora pattern where different groups show the same disjunct bigeographical pattern.
Model uncertainty in ancestral area reconstruction: a parsimonious solution?
Pirie, MD 1 , Humphreys, AM
, Linder, HP 2
2 , Antonelli, A 3 , Galley,
1
C 4
University of Stellenbosch, Dept of Biochemistry, South
Africa; 2 Institute for Systematic Botany, University of
Zurich, Switzerland;
Gothenberg, Sweden; 4
3 Gothenburg Botanical Garden,
Trinity College, Dublin, Ireland
Increasingly complex likelihood-based methods are being developed to infer ancestral areas for biogeographic inference. The results of these methods are highly dependent on the underlying model, yet techniques that take into account uncertainty not only in the estimation of the phylogenetic tree but also in the estimation of the state transition model (rates of events such as dispersals) are rarely used in a biogeographic context. Our example is the dispersal history of grass subfamily Danthonioideae (Poaceae). We infer ancestral areas and rates of dispersal using a likelihood-based
Bayesian method and show the results to be indecisive compared to those obtained under Fitch parsimony (FP), in which dispersals are minimised. With simulated data we show that at relatively low overall dispersal rates the correct root state can be inferred reliably under FP, even
279

when dispersal frequencies are unequal. Thus, despite at least 18 dispersals between the continents of the Southern
Hemisphere (over c. 30 Myr and the radiation of 280 species), and the potential for bias due to differences in distances between continents and West Wind Drift, the biogeographic scenario for Danthonioideae based on FP is robust. Only with higher rates does FP become inaccurate and likelihood based approaches more appropriate.
A Bayesian approach to estimate the biogeographyrelated diversification rates: examples from
Fosterella
(Bromeliaceae) and
Lupinus
(Fabaceae)
Silvestro, D 1, 2 , Wagner, N 3
Ibisch, P
, Schulte, K
5 , Nowicki, C 5 , Zizka, G 6
4 , Weising, K
, Schnitzler, J 2
3 ,
1 Dept of Botany and Molecular Evolution, Senckenberg
Research Institute, Frankfurt, Germany; 2 Biodiversity and Climate Research Centre, Frankfurt, Germany;
3 Plant Systematics and Morphology, Institute of Biology,
University of Kassel, Germany; 4 Australian Tropical
Herbarium, James Cook University, Cairns,
Australia; 5 Eberswalde University for Sustainable
Development, Germany; 6 Dept of Diversity and
Evolution of Higher Plants, Goethe University,
Frankfurt, Germany
Recently developed likelihood-based approaches for the reconstruction of historical biogeography along dated phylogenies have proved to outperform non-parametric methods. These novel approaches provide a statistical framework that allows the estimation of model parameters such as dispersal rates and ancestral areas, while accounting for the divergence times derived from molecular clock analysis. A combination of ancestral area reconstruction and estimated rates of speciation and extinction can lead to a better understanding of the species' present distribution. We demonstrate the potential of the method on the dated phylogenies of two genera with highly diverse clades in the Andes:
Fosterella (Bromeliaceae) and of Lupinus (Fabaceae).
We applied maximum likelihood methods as well as
Bayesian mutational mapping to reconstruct the geographic history of the genus inferring ancestral distributions, dispersal events, and range expansions. In addition we used a newly developed Bayesian approach to infer diversification rates through time as well as clade-specific speciation and extinction rates. Our method allows us to test for different hypotheses concerning the processes underlying radiation by performing model selection with Bayes factors. Tests for shifts in diversification rates found that the rates in both
Fosterella and Lupinus have been fairly stable through time. However, a clade-specific estimation of speciation rates depicted a quite different scenario, particularly showing that the diversification process is strongly correlated with the geographic distribution of the clades.
In Fosterella , we found an interesting inverse correlation between the extent of the clades' biogeographic ranges and their speciation rates. In Lupinus we observe a fourfold variation in speciation rate between the Old World and non-Andean New World lineages, while the Andean clade displays the already described explosive event of radiation. We describe how the concept of geographyrelated diversification rates can be extended to a metaanalysis approach for a better understanding of patterns
280 of species diversity resulting from speciation and extinction processes.
Evolution of gender dimorphism in Australasian
Malveae (Malvoideae, Malvaceae)
Tate, JA 1 , Skema, C
Barker, WR 3
1 , McLay, TB 1 , Wagstaff, SJ 2 ,
2
1 Massey University, Palmerston North, New Zealand;
Landcare Research, Lincoln, New Zealand; 3 State
Herbarium of South Australia, Adelaide, Australia
While most plants are hermaphroditic, gender dimorphism, with male and female functions separated in different flowers, has evolved repeatedly in unrelated families and is particularly prevalent on oceanic islands.
New Zealand has the highest reported incidence of gender dimorphism with 23% of the indigenous taxa showing some form of separate male and female function in flowers. Two prevailing hypotheses for the high frequency of this breeding system on islands have been proposed: (1) gender dimorphism evolves on islands from an initially hermaphroditic, self-compatible founder
(to promote outcrossing) or (2) gender dimorphic groups are favored to establish on islands and are most closely related to gender dimorphic taxa from the source area. To distinguish between these hypotheses, a phylogenetic framework is required. The Australasian members of
Malvaceae (tribe Malveae, subfamily Malvoideae) provide an ideal system to evaluate these alternative scenarios. The widely distributed Australian genus
Lawrencia includes both hermaphroditic (5 spp.) and dioecious (7 spp.) species. Two more narrowly distributed dioecious genera, Asterotrichion (1 sp.) and
Gynatrix (2 spp.), occur in Tasmania and mainland
Australia, and two genera are endemic to New Zealand:
Hoheria (6 hermaphroditic spp.) and Plagianthus (2 dioecious spp.). Previous phylogenetic hypotheses based on chloroplast and ITS sequence data revealed multiple origins of dioecy: one in the clade containing
Plagianthus , Asterotrichion , and Gynatrix (this clade was sister to Hoheria ) and at least one within Lawrencia .
Resolution at key nodes was inadequate to distinguish between these two alternate hypotheses. We are increasing sampling within Lawrencia and sequencing additional chloroplast markers and low-copy nuclear genes to refine phylogenetic resolution. The inferred phylogeny will be used to determine the number of times dioecy has arisen within this group and the developmental pathway by which dioecy has evolved in the island taxa.
Evolution, wood anatomy and biogeography of climbing Annonaceae
Meinke, S 1,2 , Kessler, PJA 3 , Smets, EF 3 , Chatrou, L 4
1 Hortus Botanicus Leiden, Leiden University, The
Netherlands; 2 Netherlands Centre for Biodiversity
Naturalis (section NHN), Leiden University, The

Netherlands;
Leuven,
3 Laboratory of Plant Systematics, KU
Belgium; 4 Wageningen
Biosystematics Group, The Netherlands
University,
A lianous growth habit has evolved several times within the family Annonaceae (Magnoliales, Magnoliids). Its distribution shows a biogeographic and phylogenetic bias. Whereas lianas constitute at least 30 % of the
Palaeotropical (Australasian and African) Annonaceae species, only very few climbing species can be found in the Neotropics. Apart from two small basal clades including Anaxagorea and Ambavia , the Annonaceae are classified into two major clades: malmeoids and annonoids, previously called short branch clade and long branch clade respectively. Lianas only evolved in the annonoids, and can be found amongst others in the genera Desmos , Fissistigma , Friesodielsia ,
Monanthotaxis , and Uvaria . These genera are part of a large clade of climbers consisting of 14 genera and about
400 species, forming the so called ‘climber clade’.
Another species-rich (125 spp.) clade is made up only by the palaeotropical genus Artabotrys which has a unique climbing device. Additionally, at least 3 more origins of climbers can be recognized within Annonaceae, however, each with only very few species. This study is part of a
PhD project on the evolution and distribution of the climbing habit, using Annonaceae as a model group. In this paper three topics are raised. First, the definition of the term liana (within Annonaceae) will be addressed.
This is problematic in Annonaceae as many intermediate growth forms can be found, e.g. several species start growing as shrubs and only at later stages develop into a climber. This problem will be approached using wood anatomical characters of the different climber types.
Second, using molecular phylogenetic studies based on the chloroplast markers trn L-F, ndh F, psb Atrn H, atp Brbc L and trn S-G, conclusions are drawn about the number of origins of the climbing habit within the family and on possible reversals to a non-climbing habit.
Furthermore, an overview on the biogeography of climbing Annonaceae will be given.
Phylogenetic and morphological patterns of diversification in the Australian Proteaceae
Miller, E 1 , Zanne, A 2
1 Macquarie University/U. Missouri, St Louis, USA; 2 U
Missouri-St Louis, St Louis, USA
Phylogenetic community structure is a measure of the manner in which the species in a community are arrayed across a phylogeny that includes both those species and other related species from the region. Such measures can vary from clustered, where the species in a community come from a restricted subset (or subsets) of the phylogeny in question, to overdispersed, where the species are less closely related than expected by chance.
Australia is characterized by an extraordinarily diverse
Proteaceae flora. These species are diverse in sheer number and in morphology. Yet, many of the species are restricted to a small subset of the larger Proteaceae phylogeny, in particular groups such as the Hakeinae,
Banksiinae, and the Persoonioideae. What are the patterns that characterize the Proteaceae radiation across the Australian continent? We quantify the phylogenetic community structure of the Proteaceae along rainfall and temperature gradients, and show that communities become ever more phylogenetically clustered along gradients of decreasing temperature and precipitation.
When we focus on smaller clades within the Proteaceae, however, the opposite patterns are sometimes observed.
The Hakeinae, for instance, are more phylogenetically clustered in wetter areas, which highlights their well known affinities for arid environs. We also discuss to what degree morphology, e.g. plant functional traits, has varied throughout the course of this diversification. Much of the morphological diversity observed across Australia is focused in the aforementioned speciose but phylogenetically restricted subset of the Proteaceae. Is this simply a matter of random character evolution that manifests itself as a large diversity of life forms when such a large sample of species are considered, or can, for instance, these shifts be attributed to competitive interactions and/or the filling of unoccupied niches in colder and drier communities? What morphological traits, if any, are associated with these shifts in environmental space? We present preliminary analyses on these latter questions, and discuss the implications for how the processes may have lead to the patterns of phylogenetic community structure we observe along rainfall and temperature gradients.
Speciation and phylogeography of
Telopea
and
Lomatia
(Proteaceae)
Milner, M 1,2 , Crisp, MD 1 , Rossetto, M 2 , Weston, PH 2
2
1 Australian National University, Canberra, Australia;
National Herbarium of NSW, Botanic Gardens Trust,
Sydney, Australia
Australia is a unique continent with a climatic past that has resulted in widespread aridification and the evolution of dry, sclerophyllous vegetation. Changes in climate and landform have transformed the Australian flora resulting in the diversity and distribution present today. Using phylogeographical methods we can test genetic patterns for evidence of a changing landscape, such as vicariance events, retraction into refugia and consequential reexpansion, and adaptation to a new environment.
Lomatia and Telopea (Proteaceae) are sister groups that, within Australia, display almost identical overall distributions in eastern Australia, making them ideal for exploring speciation processes. The overlap in distribution suggests that if historical events, such as climate change, have led to allopatric speciation, this should have resulted in congruent geographic patterns in both genera. Using nuclear and chloroplast sequences, we will investigate differences and similarities of genetic structure across and within these genera, to understand the major contributors to speciation, distribution and gene flow. Discovering the processes dominant within
Lomatia and Telopea will assist in determining what historical events have been important for the speciation of eastern Australian flora.
Associations of Australasian plants and animals through the Cenozoic
Cook, LG 1 , Hardy, NB 2
281

1 The University of Queensland, Brisbane, Australia;
2 Queensland Primary Industries and Fisheries, Brisbane,
Australia
Understanding the processes that have generated the world’s biodiversity is a major challenge of evolutionary biology. The interactions among organisms are thought to be among the major drivers that creates and maintains species diversity. A great diversity of insects and plants depend on one another for food and pollination, and their interactions affect population persistence, extinction and diversification. Together, insect-plant interactions likely form the basis of much of the world’s current biodiversity. Worldwide, there is evidence that major geological and climatic events, such as at the end of the
Cretaceous, caused concerted extinction via linkages in food webs and interaction networks. That is, even species that were not directly affected by climatic changes ultimately went extinct as follow-on effects cascaded through interaction webs. After such disruptions, it can take many millions of years for interaction webs to establish similar levels of connectedness. Although the fossil record shows a long and rich record of plantherbivore interactions, it generally lacks the resolution to help address questions about specific associations.
Comparative analyses of dated molecular phylogenies are now contributing much to our understanding of associations through time. Given that plant distributions appear to be, at least in part, evolutionarily constrained by the inability of plants to change niches, interactions between plants and insects, at the broad scale, are likely also limited by phylogenetic history. In interacting organisms, such as plant-feeding insects and their host plants, population dynamics (including dispersal, distribution, and genetic and phenotypic diversity) of the host directly affect those of the insect. However, despite early predictions by Ehrlich and Raven that specific herbivore-plant interactions should co-evolve, few, if any, studies have found evidence of long-term coradiations (a prediction of long-term coevolution).
Perhaps plant-herbivore interactions are more dynamic than first presumed, or long-term associations require certain environmental criteria not experienced by most interacting taxa studied to date. Given that the fossil record shows decoupling of insect-plant interactions following major extinction events and climate change, it might be expected that long-term associations are more likely in the most stable conditions. To date, the majority of tests for plant-herbivore co-radiation have centred on systems in the Northern Hemisphere, which has likely suffered dramatic climate change through the Plio-
Pleistocene periods with glacial cycles. The Australian landmass has been largely isolated since its separation from Antarctica between 30-50 million years ago. As a consequence, much of the flora and fauna present from that time has evolved relatively independently, with little interaction with other world biotas until recently.
Furthermore, Australia did not undergo extensive glaciation during the Pleistocene. We will examine whether these environmental factors have contributed to the evolution, and long-term persistence, of plantherbivore associations. We will address the question of whether there has been co-radiation between plants and herbivores in Australasia and whether interactions, in general, might be partly explained by levels of stability, isolation and host abundance or richness.
282
Regional mass extinction of Australian sclerophyll
1 species in the Pleistocene
Sniderman, K 1 , Jordan, G 2
School of Geography and Environmental Science,
Monash University, Australia; 2 School of Plant Science,
University of Tasmania, Australia
The sclerophyll floras of the Cape Floristic Region in
South Africa and the South West Floristic Region in
Australia represent the most diverse floras outside of the tropics. Although these hyperdiverse biomes are part of the globally rich mediterranean climate systems, it has been understood that their origins may predate the onset of their winter wet/summer dry climate regimes.
However, there has long been a view that sclerophyll biota radiated, and mesic rainforest biota declined in these regions, in response to late Neogene climate changes. Here, we use a fossil leaf flora to demonstrate the presence of a hyperdiverse sclerophyll flora in southeastern Australia in the Early Pleistocene.
Taphonomic modelling suggests that the source vegetation was at least as rich in woody species as the most species-rich extant sclerophyll floras globally, and much richer than any sclerophyll vegetation in eastern
Australia today. Our results indicate that climaticallydriven winnowing of the southeastern Australian sclerophyll flora was of a similar magnitude to that of the rainforest flora, which experienced a catastrophic decline in response to the onset of high-amplitude glacial climate cycles at around ~1 million years ago. We attribute the loss of hyperdiverse sclerophyll floras in eastern
Australia to processes described by Jansson and
Dynesius (2000, Annu. Rev. Ecol. Syst. 33:741), in which rapid and extreme Pleistocene climate fluctuations progressively culled specialists, leaving a smaller set of generalist species. Our results suggest that hyperdiverse floras have persisted in the the South West Australian
Floristic Region either because the amplitude of
Pleistocene glacial climate cycles has been less than in the southeast; and/or because a large proportion of southwest species were pre-adapted to the types of demographic change induced by glacial climate cycles.
Phylogenetic relationships, tempo and patterns of evolution of the Australasian Ericaceae
Puente-Lelièvre, C
1
C 3 , Brown, E 3
1 , Harrington, M 1
, Crayn, D 1
, Hislop, M 2 , Quinn,
Australian Tropical Herbarium, James Cook University,
Cairns, Australia;
Perth, Australia; 3
2 Western Australian Herbarium,
National Herbarium of New South
Wales, Royal Botanic Gardens Sydney, Australia
The subfamily Styphelioideae Sweet, Ericaceae Juss.,
(the epacrids) includes 35 genera and some 420 species distributed mainly throughout Australasia with outliers extending the range to Hawaii and Tierra del Fuego
(Chile). They are usually perennial woody shrubs ranging in size and habit from prostrate forms to forest trees.
Their habitat varies between heathlands, sandplains to tropical forest. The Stypheliodeae are particularly diverse in Australia and are regarded as an important component of Australian flora particularly in heathlands. Generic circumscription in Styphelioideae has been challenging

since the time it was first described by Brown as the family Epacridaceae. In order to resolve these taxonomic problems, molecular phylogenetic studies have been undertaken on Styphelioideae. The result has been resolved and corroborated phylogenetic trees that outline the major clades of the subfamily and have lead to a more detailed revisionary work on a clade-by-clade basis.
This study focuses on the tribe Styphelieae, the largest tribe in Styphelioideae, where non-monophyletic genera such as Leucopogon R.Br., Astroloma R.Br. and
Styphelia Sm. persist. Molecular data combined with morphological characters have identified new groups supported by molecular and morphological synapomorphies. Many of these new groups appear to be biogeographically based and may warrant recognition at generic level. The species of Styphelia from New
Caledonia form one clade; and distinct Western
Australian and eastern Australian groups are resolved.
The epacrids typify many of the controversies in the historical biogeography of Australasia. Their distribution is almost exclusively restricted to Gondwanan fragments
(Australia, New Zealand, New Caledonia, and South
America with outliers extending the range to Hawaii and some Pacific islands). They are woody plants growing mostly in heaths and montane forests. These ecological traits are considered to decrease the probability of transoceanic dispersal. Conversely, their fleshy fruit and small seeds should increase this probability. We discuss the evolutionary processes that may have contributed to their current distribution. Fossil-calibrated Bayesian relaxed molecular clock analyses indicate that the minimum age of the tribe Styphelieae is 22-25 Ma.
According to this estimation, the minimum ages of the south-western versus south-eastern Australian disjunctions is between 11 to 7 Ma. The fact that these disjunctions show temporal clustering suggests vicariance, and leads to the conclusion that components of the tribe Styphelieae represent an example of southwest and south-east Australian biotas that were isolated during the mid-Miocene aridification and the elevation of the Nullarbor Plain. We also document long-distance dispersal events between Australia, Papua New Guinea, and New Zealand in the genera Leucopogon and
Acrothamnus.
Analysis and interpretation of historical biogeographic patterns in continental Australia
Ladiges, P 1 , Bayly, M 1 , Nelson, G 1
1 School of Botany, The University of Melbourne,
Australia
Many modern biogeographic studies based on molecular taxon phylogenies, invoke the Progression Rule, based on the ideas of Willi Hennig and Lars Brundin. It is assumed that the most primitive or basal taxa are likely to occur at the centre of origin, indicating the ancestral area of the group. Progression from the basal node to the most derived is interpreted as the direction of dispersal of the group. The Progression Rule is invoked where there is geographic paralogy (duplication of biotic areas in the phylogeny). An alternative approach is to recognise and eliminate geographic paralogy. The present-day distribution of the whole group can be considered largely to represent the ancestral area and the taxon phylogeny and distribution of endemics then used to discover any pattern of biogeographic differentiation. Interpretation of the history of the biogeographic pattern may invoke various causes and processes: vicariance, long-range dispersal, short-range expansion, contraction or extinction, driven by geological, climatic or other changes. Geographic paralogy is illustrated by the phylogenetic position of taxa that are endemic to South
West Western Australia and have relatives in other regions of continental Australia. The South West bioregion, with its high level of endemicity, is not interpreted as a centre of origin but as one of the earliest bioregions to differentiate as Australia rifted from
Antarctica; the South West became isolated by marine inundation across southern Australia and by climatic cooling since the Late Eocene–Early Oligocene, processes that led to the isolation and differentiation of clades from west to east.
Spinifex grasslands of Australia’s arid zone: past and future uses
Gamage, H 1
Memmott, P 1
, Wallis, L 1
, Schmidt, S 1
, O’Rourke, T 1 , Martin, D 1 ,
1 University of Queensland, Australia
Spinifex grasslands, dominated by ~70 endemic species in the genus Triodia , are the foremost vegetation type of
Australia’s arid zone and cover approximately one-third of the continent. Spinifex grasses are extremophiles which inhabit some of the hottest and driest conditions on earth; they display distinctive morphological and physiological adaptations for survival in environmental extremes. Spinifex grasslands resemble shrublands owing to their long-lived hardy foliage and hummock structure. The biomass of mature spinifex grasslands can reach 15 tonnes per hectare which is high compared with semi-arid regions globally. Aboriginal fire regimes have shaped spinifex grasslands for 50, 000 years. The spinifex fire cycle commences with numerous herbaceous species being present and concludes after 5 to
30 years with spinifex remaining as the dominant vegetation. Traditional anthropogenic burning regimes resulted in a mosaic of vegetation with different fire histories that enhanced hunting efficiency, biodiversity and prevented uncontrolled wild fires. In traditional
Aboriginal culture, spinifex biomass and resin were used extensively purposes that include shelter cladding and hafting of tools and weapons. In the post-colonial era, much of Australia’s spinifex grasslands have been used for cattle grazing, facilitated by promoting exotic grasses.
Invasion by exotic plants and wildfires threaten the integrity of spinifex grasslands. The large extent of spinifex grasslands and the changes associated with altered land use, fire regimes and climate, raise questions as to how best manage spinifex grasslands. Indigenous knowledge informs potential novel uses of Triodia spp., such as thermal insulation for buildings and composite materials and adhesives; potentially providing sustainable economic pathways for desert communities.
Using both an Indigenous and Western scientific framework, research is underway to evaluate the material properties of spinifex biomass and resin, and the effects of harvesting on ecological processes. Here, we present a review of spinifex grasses in the context of biology,
Indigenous ethnobotany and material science.
283

Hybridization and introgression across different ploidy levels in the Neotropical orchids
Epidendrum fulgens
and
E. puniceoluteum
(Orchidaceae): insights from reproductive compatibility, population genetics and cytogenetics
Pinheiro, F
Fay, MF 3
1 , Palma-Silva, C 1
, Cozzolino, S 4
, Moraes, AP 2 , Barros, F 1 ,
1 Instituto de Botanica, Sao Paulo, Brazil; 2 Instituto de
Biologia, Universidade Estadual de Campinas,
Campinas, Brazil; 3 Jodrell Laboratory, Royal Botanic
Gardens Kew, UK; 4 Universita degli Studi di Napoli
Federico II, Dipartimento di Biologia Strutturale e
Funzionale, Napoli, Italy
The hypothesis of gene flow between species with large differences in chromosome numbers has rarely been tested in the wild, mainly because species of different ploidy are commonly assumed to be reproductively isolated from each other because of instantaneous and strong postzygotic barriers. In this study, an interdisciplinary survey based on population genetics, phylogeography and cytogenetics was carried out in two orchid species with different ploidy levels: Epidendrum fulgens (2n = 2x = 24 chromosomes) and Epidendrum puniceoluteum (2n = 4x = 52 chromosomes). To test the strength of their reproductive barriers, we investigated the distribution of genetic variation (nuclear and plastid microsatellites) in sympatric and allopatric populations across the geographical range of both species along the
Brazilian coastal plain. In addition, genome compatibility and fertility of E. fulgens , E. puniceoluteum and the hybrid were estimated by: (a) testing seed viability of artificial crosses; (b) analyzing meiotic behavior of pollen mother cells; and (c) counting chromosome numbers. All six sympatric populations analyzed presented hybrid zones, indicating that hybridization between E. fulgens and E. puniceoluteum is a common phenomenon. Bayesian assignment analysis detected the presence of F1 and F2 individuals and also signs of introgression, indicating a high potential for interspecific gene flow. Introgression occurs preferentially from E. fulgens to E. puniceoluteum . Pure parental individuals of both species display strong genotype–habitat associations, indicating that environment-dependent selection could be acting in all hybrid zones. Artificial crosses showed that the two parental species produced fruits with highly viable seeds in intraspecific and interspecific crosses. F1 hybrids and backcrosses towards
E. puniceoluteum only produced fertile seeds when they were pollinated by pure E. puniceoluteum plants in agreement with assimetrical patterns of introgression revealed by Bayesian assignment analysis. The cytogenetic analyses confirmed parental chromosome numbers and revealed and unexpected chromosome number of n = 19 (2n = 38 chromosomes) for hybrid individuals. A regular meiotic behavior was observed in parental species, whereas in hybrids, meiotic behavior was irregular, indicating low viability and pointing to the presence of post-zygotic barriers that maintain cohesion of species’ gene pools in hybridizing populations. Our
284 data suggests that hybridization and introgression are evolutionary processes playing a role in the diversification of the genus Epidendrum . The present scenario indicates the importance of investigations of hybrid zones to understand the role of reproductive barriers and speciation processes in Neotropical orchid species.
Addressing the flow cytometric puzzle: a remarkable coexistence of multiple cytotypes in the Fragrant
Orchid (
Gymnadenia conopsea
agg.)
1
Kubátová, B
Krajníková, E 1
1 , Trávníek, P 0
, Jersáková, J 4
, Urn, V
, Suda, J 0
1 , Rauchová, J 0 ,
University of South Bohemia ineské Budjovice, Faculty
2 of Agriculture, Biotechnological Centre, Czech Republic;
Academy of Sciences of the Czech Republic, Institute of
3
4
Botany; Charles University in Prague, Faculty of
Sciences, Deparment of Botany, Czech Republic;
University of South Bohemia ineské Budjovice, Faculty of Sciences and Institute of System Biology and Ecology,
Czech Republic
One of the prerequisites for polyploid research in natural systems is knowledge of the geographic distribution of cytotypes. In our study, we examined inter- and intrapopulational ploidy diversity in the fragrant orchid in Central Europe and searched for potential explanations and evolutionary consequences of the observed spatial pattern. Gymnadenia conopsea (L.) R.Br. s.l. (fragrant orchid\ Orchidaceae) is an Eurasian species that shows considerable morphological, phenological and/or karyological variation across its distributional range. Five different ploidy levels (4x, 6x, 8x, 10x, and 12x) were detected among 3577 plants analyzed by DAPI flow cytometry, with hexaploids being recorded for the first time. Tetraploid and octoploid cytotypes clearly predominated and accounted for 61.8% (37.6% corresponding to G. conopsea s.s. and 24.2% corresponding to G. densiflora ) and 35.6% of all plants, respectively. The frequency of minority ploidy levels
(6x, 10x, and 12x) varied from ~1.9% in hexaploids (67 individuals) to ~0.5% in decaploids (18 individuals) and
~0.2% in dodecaploids (nine individuals). The scarcity of minority cytotypes suggests the existence of strong pre- or postzygotic mating barriers, the nature of which remains to be determined. Although minority cytotypes accounted for only 2.6% of all samples, they were found in more than half of the studied populations (22 out of
43). Ploidy mixing was very common and more than
55% of analyzed Gymnadenia populations (24 out of 43) harboured two or more cytotypes. A certain spatial structure was observed in plots laid out in populations with the highest cytotype variation, including clumping of individuals of the same ploidy and negative association between tetra- and octoploids. Gymnadenia tissues were found to undergo a progressively partial endoreduplication, which accounts for ~60% and ~75% of the total genome in G. conopsea and G. densiflora , respectively. Progressively partial endoreduplication is a rare phenomenon of general biological significance, which has only been observed in orchids. Collectively,
Gymnadenia conopsea agg. represents a remarkable example of a large intraspecific ploidy variation coupled with a frequent sympatric growth of several (up to five) different cytotypes. As compared to other thoroughly

investigated polyploid complexes, the Fragrant Orchid is a more complicated system, which can provide novel insights into the mechanisms and dynamics of polyploid speciation under natural conditions.
Where have the hexaploids vanished? The role of breeding barriers in coexistence of
Gymnadenia conopsea
cytotypes
Castro, S 1 , Jersáková, J 2
Kubátová, B 2 , Urn, V 2
, Dötterl, S 3 , Trávníek, P 4 ,
1 University of Coimbra, Portugal; 2 University of South
Bohemia, Ceske Budejovice, Czech Republic; 3 University of Bayreuth, Germany; 4 Faculty of Science, Charles
University in Prague, P Czech Republic
Polyploidy has played a key role in plant diversification.
Despite this, it is still largely unknown how various factors contribute to reproductive isolation among cytotypes in mixed-ploidy populations. Our study aims at investigating the origin and maintenance of contact zones of various cytotypes, using Gymnadenia conopsea complex (Orchidaceae) as study model. Specifically, we assessed cytotype distribution in Central Europe and investigated pre- and post-zygotic barriers among cytotypes in mixed-ploidy populations. Using flow cytometry, we determined DNA ploidy levels in 43 populations revealing two majority (4x, 8x) and three minority (6x, 10x, 12x) cytotypes, often in mixed-ploidy populations. To investigate pre-mating barriers, we determined cytotype spatial distribution, flowering phenology, floral morphology, and visual and olfactory cues, which could lead to assortative mating. Gas chromatography coupled with electroantennographic detection was used to identify scent compounds with physiological activity in the two main pollinators,
Deilephila porcellus and Autographa gamma . The importance of olfactory cues was also assessed in the field by analysing the moths’ responses to the olfactory display of the plants, and by following the pollinator’s behaviour on artificial arrays. Artificial crosses were used to asses post-mating barriers. Our complex approach demonstrated that the coexistence of
Gymnadenia cytotypes in mixed-ploidy populations is partly maintained by differences in floral phenology and spatial segregation of the cytotypes. However, cytotypes with overlapping flowering can freely hybridize due to similarities in floral colour and spur length, the lack of assortative behaviour of pollinators and the absence of gametic incompatibility. Though both pollinator species seemed to physiologically detect differences in floral scent, and the scent alone was sufficient to attract them, pollinators did not use this cue to discriminate the cytotypes in the field. Crossing experiments revealed that minority ploidies largely originate from crosses between
4x and 8x plants, with most progeny being 6x, whereas backcrossing of 6x with parents results in very low seed quality. The co-occurrence of dominant cytotypes in mixed populations is thus maintained by temporal and spatial segregation and limited reproduction of hybrids.
The absence of a barrier to pollen discounting as well as the scarcity of 6x plants is discussed.
Non-random distribution of orchids reflects different mycorrhizal association patterns in a hybrid zone of three
Orchis
species
1
Jacquemyn, H 1
University of Leuven, Belgium
The abundance and distribution of plant species are determined by a wide range of factors, including localized dispersal, intra- and interspecific interactions and heterogeneous environmental conditions. Because in orchids seed germination is critically dependent upon the availability of suitable mycorrhizal fungi, their spatial distribution and abundance are therefore likely to be affected by the way mycorrhizal fungi are distributed in natural populations. To test the hypothesis that the spatial distribution of orchid species reflects associations with mycorrhizal fungi, we examined spatial aggregation patterns and mycorrhizal associations in a hybrid zone containing three species of the genus Orchis ( Orchis anthropophora , O. militaris and O. purpurea ). We first investigated the extent of hybridization in this population using molecular markers and Bayesian assignment tests.
Spatial point pattern analyses were then combined with seed germination experiments using both pure and hybrid seeds to investigate the spatial extent of hybridization in this population and how this was affected by interactions with mycorrhizal fungi. Hybridization between O. anthropophora and the other two species was limited. In contrast, O. militaris and O. purpurea hybridized frequently within this population. However, despite frequent hybridization between O. militaris and O. purpurea , significant spatial clustering of the three species and hybrids was observed. Assessment of mycorrhizal associations in both pure and hybrid plants showed that mycorrhizal specificity was low, but each species associated with a particular set of fungal lineages.
Consistent with the results of the spatial point patterns analyses, the community of fungal lineages associating with hybrid plants and pure O. purpurea individuals, was very similar, suggesting that the spatial structure of both pure and hybrid plants are to some extent sensitive to the contingencies of the spatial distribution of suitable mycorrhizal fungi. The integration of spatial point pattern analysis, molecular analysis and the analysis of mycorrhizal associations in both pure and hybrid individuals allowed assessing the spatial extent of hybridization and elucidating the factors generating spatial clustering in three co-occurring orchid species.
A population glimpse into the emergence of heterotrophy in orchids: genetic and ecophysiological investigations on achlorophyllous (albino) variants in
Cephalanthera damasonium
Selosse, M-A 1 , Roy, M 1 , Damesin, C 2 , Berveiller, D 2
1 Centre d'Ecologie Fonctionnelle et Evolutive,
Montpellier, France; 2 Laboratoire d'Ecologie,
Systématique et Evolution, Université Paris-Sud, France
Among green plants, achlorophyllous individuals (= albinos) usually do not survive. There are, however, some exceptions in green orchid species that are phylogenetically related to fully heterotrophic species.
The later use their mycorrhizal fungi as carbon source
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(mycoheterotrophy), and the former use both autotrophy and partial mycoheterotrophy (mixotrophy). In this context, albinos likely survive as true mycoheterotrophs, as supported by their natural isotopic abundances: albinos, together with fully mycoheterotrophic orchids, are richer in 13 C and 15 N than autotrophic plants, exactly as their mycorrhizal fungi. Mixotrophic individuals show intermediate isotopic abundances. Although green individuals and albinos look morphologically similar, the albino frequency in populations is typically <5%, and their demography is lower than that of albinos. To understand what makes albinos less fit, we engaged a survey of multiple vegetative and reproductive parameters on a French population of Cephalanthera damasonium (Neottieae), where a high frequency of albinos allowed some statistical comparison over 3 years.
Using AFLP fingerprints albinos showed not more genetically related among them than with green individuals; furthermore, they do not cluster spatially in the population, so that they may reproduce. Like green individuals, albinos sometimes stayed underground during one or two years (‘dormancy’) but did so more often. A monitoring of shoots showed that phenology was synchronous for growth, flowering and fruiting. No significant difference in morphology was detected, but at fruit maturation, albinos more often went dry, leading to fruit abortion. Using isotopic abundances over a growth season, we showed that mixotrophic shoots shifted from nearly mycoheterotrophic at shoot emergence to very autotrophic later in the season, when fruit ripened.
Accordingly, in situ measurements of gas exchanges showed an increase in photosynthesis over the season; conversely, mycorrhizal colonization decreased over the season. Albinos were thus probably carbon-limited at ripening. Two additional factors may act: (i) fungal and insect pathogen loads were higher for albinos, perhaps due to their less mimetic habit and/or their higher N content (a feature expected in mycoheterotrophs); (ii) temperature measurements and stomatal response to vapour pressure deficiency supported that albinos could undergo over-heating at high temperatures (>30°C).
Thus, carbon nutrition, biotic factors and over-heating may all increase shoot drying in albinos. We also investigated the fitness of both phenotypes: although fertilization was equally frequent, fruits were smaller and had less seeds that germinated less frequently after two years in soil. Controlled crosses supported that this was a maternal, not a paternal effect, so that the previous factors may also explain these trends. This case study highlights that albinos have a 40X-reduced fitness and are cryptopolymorphic. It supports a contrario that several physiological and morphological traits should coevolve in mixotrophs to achieve a successful mycoheterotrophy. We will list them, and claim that their co-evolution is a major limitation to the shift to full mycoheterotrophy in adult orchids, although they are able to use fungal carbon, at germination or as mixotrophs.
Mycorrhizal fungi played key roles in the evolution of orchids
Yukawa, T 1 , Ogura-Tsujita, Y 1
1 Tsukuba Botanical Garden, National Museum of Nature and Science, Japan
The Orchidaceae, the largest plant family, comprises about 25,000 species. The greater part of the diversification in the family occurred in sites of high stresses on water, light (intense or dark), and/or nutrients.
High percentages of the epiphytic species (60% of vascular epiphytes) and the mycoheterotrophic species
(40% of vascular mycoheterotrophs) in orchids illustrate well this characteristic. Further, an important character related to nutritional innovation, the obligate mycoheterotrophy during the juvenile stage, represents a synapomorphy of orchids because this feature is not found in closely related groups to orchids. We hypothesized that nutritional contributions from mycorrhizal fungi facilitated niche expansion and subsequent radiation of orchids in highly stressed sites where competition is expected to be low. To test this hypothesis, we analyzed (1) the origin of specialized relationships between orchids and fungi and (2) the correlation between fungal and orchid diversification.
The first issue was examined by fungal symbiosis of
Apostasia , which is the earliest-diverging clade of orchids in which mycorrhizal associations are scarcely known. Mycobionts of Apostasia form typical orchid mycorrhizae and the results of molecular identification showed that they belong to the order Cantharellales of
Basidiomycota. Most other groups of orchids are also associated with this group, while the sister group and other closely related groups to Orchidaceae ubiquitously form symbioses with Glomeromycota to form arbuscular mycorrhizae. This indicates that a major shift in fungal partner occurred in the common ancestor of the
Orchidaceae. The second issue, correlation between fungal and orchid diversification, was tested by
Cymbidium . The genus exhibits distinctive ecological diversification such as terrestrial, epiphytic, and lithophytic life forms. Moreover, a few species evolved obligate mycoheterotrophy. Molecular identification of mycorrhizal fungi in Cymbidium revealed that the fungal groups shifted in accordance with the evolution of life forms (epiphyte to terrestrial) and nutritional modes
(autotrophy to mycoheterotrophy). These results demonstrated that relationships with particular groups of mycobionts played key roles in the enormous diversification of Orchidaceae.
Evolution of the Eurasiatic genus
Himantoglossum
(Orchideae, Orchidoideae): an integrative
1 phylogenetic approach
Sramko, G 1 , Molnar, AV 1 , Hawkins, J 2 , Bateman, R 3
2
Dept of Botany, University of Debrecen, Hungary;
School of Biological Sciences, University of Reading,
Berkshire, UK; 3 Jodrell Laboratory, Royal Botanic
Gardens Kew, UK
Evolutionary relationships among taxa of the
'Steveniella'– Himantoglossum ' clade, with particular interest in the endangered genus Himantoglossum s.str., has been reconstructed using morphometric and molecular phylogenetic analyses. The sample set included all taxa of the clade covering its whole distribution range. A total of 46 morphometric characters measured in the field were analysed using multivariate techniques to characterise the morphological variability and circumscribe morphological entities. Molecular phylogenetic methods, utilising moderately and fast
286

evolving cpDNA ( acc Dpsa I, atp Fatp H, rps 16, psb Atrn H, trn Lrpl 32ndh F) and nuclear markers (nrITS and
LFY), were sequenced and used in phylogenetic tree reconstruction to describe and interpret the evolution of the group. Species from eastern Europe and Asia Minor formed the basal branches of the group, while a western expansion can be concluded from the tree topologies; the
Himantoglossum s.s. species were placed on the most recently diverging branches. Sister to all
Himantoglossum s.s. taxa is the isolated Azerbaijani endemic H. formosum , whereas the remaining taxa suggested more recent radiation. The morphological groups and accepted species boundaries largely corresponded with the molecular clades. Although H. hircinum and H. adriaticum formed well-supported, easily distinguished clades, the Eastern taxa ( H. caprinum s.l. and H. affine ) – though strongly separated from the above two species – were much closer to each other in terms of genetic distance. Data from genic regions of contrasting inheritance allowed the hypothesis that H. adriaticum had a hybrid origin to be tested. The detailed floral ontogeny and allometry of
Himantoglossum offer the prospect of achieving a deeper understanding of character evolution within the lineage.
Together, the complexity of the genetic patterns evident in clades of European orchids studied by us
( Himantoglossum , Ophrys , Dactylorhiza , Gymnadenia ,
Platanthera ) have encouraged exploration of contrasting methods of analyzing ITS alleles and plastid haplotypes, in order to extract maximum systematic and phylogeographic inference. One area of particular interest is a genetic divide that approximates the
Carpathians and is suggested by each of our study groups. Himantoglossum , together with the above genera, share this evolutionary history: these particular members of European Orchidinae pursued a westward migration and associated diversification.
Phylogeny and species delimitation of the Australian endemic
Corysanthes clade (genus
Corybas
,
Orchidaceae)
1
Lyon, S 1
University of Wisconsin, USA
The terrestrial orchid genus Corybas (subfamily
Orchidoideae: tribe Diurideae: subtribe Acianthinae) contains about 140 described species and ranges from
India and China through southeast Asia to New Guinea,
Australia, New Zealand and several Pacific and
Subantarctic islands. Individuals are just a few centimeters tall, with a single leaf and a single, structurally complex flower. Despite being one of the most distinctive orchid genera, they are often overlooked due to their diminutive size and have not been well studied. One clade, elevated to the genus Corysanthes by
Jones et al., is endemic to Australia. Habitat preferences in this group range widely, from coastal sand dunes to temperate rain forest and from elevations at sea level to
900 meters. Of the 12 species currently recognized, only four were described prior to 1970 and species boundaries are uncertain in many cases. Morphological variation in the C. diemenicus complex has led to recent segregation of two new species, but whether these represent distinct taxa is contested. Hybridization between the members of the C. diemenicus complex and C. incurvus has been reported. The federally endangered C. dentatus co-occurs with these taxa and has an intermediate floral morphology–hybrid origin is suspected, but has not previously been tested using molecular data. To address these issues, and to reconstruct the history of diversification in floral morphology and habitat preference, we have analyzed sequences from several hypervariable chloroplast spacer regions, the nuclear ribosomal internal transcribed spacer (ITS), and lowcopy nuclear markers. All described species apart from
C. grumulus were included, with up to five populations per species. These data reveal relatively little genetic variation within the clade, suggesting a recent radiation.
The two taxa endemic to Western Australia, though morphologically similar to species in the southeast, are clearly genetically distinct. However, our sequence data fail to resolve the issues surrounding the C. diemenicus complex, its purported hybridization with C. incurvus , and the nature of C. dentatus . The lack of resolution necessitates using a faster evolving source of molecular data. We present preliminary results from amplified fragment length polymorphisms (AFLPs), and explore their utility for further analyses of this group.
Relationships within and among species of
Spiranthes
, a cosmopolitan orchid plagued by high levels of
1 allopolyploidy and taxonomic inflation
Cameron, K 1 , Dueck, L 2 , Aygoren, D 1
University of Wisconsin – Madison, USA; 2 Athens, TX,
USA
The ladies’ tresses orchids, genus Spiranthes , are native to temperate parts of Central America, Europe, Asia, and
Australia, but are most diverse in North America where they are among the most diverse and commonly encountered of orchids. In spite of this, there exists considerable uncertainty regarding their patterns of hybridization, phylogeny, conservation status, and species delimination. For example, of the 34 currently recognized species, only 19 were known prior to 1930; the remaining 15 species and more than a dozen varieties are relatively new to the botanical literature. This is a cosmopolitan genus with a questionable systematic past that has almost certainly been subjected to excessive taxonomic inflation. To understand better the systematics of these orchids, we have generated a multi-gene hypothesis of phylogenetic relationships among >200 accessions of Spiranthes using nuclear and plastid data.
All but a few rare species were included in the analyses, and in most cases individuals from multiple populations
(as many as 25 in some cases) were included to represent the range of geographic distribution and morphology, especially within the S. cernua complex. Cladograms are generally well resolved, and most relationships receive high jackknife support values. All Old World endemic species form a clade embedded within the tree, indicating their origin from a New World ancestor. The data fail to discriminate fully the various taxa that comprise the autumn flowering S. cernua complex, and even indicate that some taxa belonging to this group (e.g., S. parksii ) may not warrant species status. AFLP data may prove useful as we explore this group further. Conversely, these molecular data have been powerful in identifying at least one new species, S. stellata , from the NW USA.
Chromosome number correlates well with the
287

phylogenetic pattern revealed (e.g., those species based on n=22 are sister to the remainder of the genus based on n=15). However, the common occurrence of allopolyploids within Spiranthes presents a challenge to understanding its evolutionary history. Continued studies, especially at the population level, will be essential for better understanding these temperate orchids.
Genetics as a tool for elucidating relationships at the species boundary – examples from European orchids
Fay, MF 1
1 Royal Botanic Gardens, Kew, UK
Genetic interactions between different species of plants can lead to confusion in the definition of what constitutes the species in question, and an increasing number of genetic techniques are being developed which allow us to probe these interactions. Genetic studies of a range of terrestrial orchids in Europe have revealed widespread occurrence of hybridization between individuals of the same ploidy (with or without a resulting change in ploidy) and between species of different ploidy, introgression (resulting from repeated backcrossing to one parental species) and the existence of cryptic species, all of which can lead to misidentifications. Examples will be provided of each of these taken from orchid genera including Orchis , Dactylorhiza and Cephalanthera . In
Orchis , populations thought to represent pure species frequently show signs of introgression from related species, and hybridization at the diploid level is frequent.
In Dactylorhiza , hybridization between diploids with accompanying allopolyploidization is a major evolutionary pattern, but hybridization across ploidy levels is also widespread. In Cephalanthera , there are potential cryptic species, but these are more difficult to assess because the large nuclear genome size of species in this genus hampers genetic fingerprinting based on nuclear DNA. Consequences for taxonomy and conservation of the results presented will be discussed.
Transcriptomic and epigenetic alterations after recurrent allopolyploidization triggers adaptive diffusion in
Dactylorhiza
(Orchidaceae)
Chase, MW 1 , Paun, O 2 , Bateman, RM 1 , Fay, MF 1
1 Royal Botanic Gardens, Kew, Richmond, UK; 2 Dept of
Systematic and Evolutionary Botany, University of
Vienna, Austria
Hybridization and polyploidy are potent forces that have regularly stimulated plant evolution and adaptation.
Hybridization with or without a ploidal shift can quickly result in alterations of gene expression and epigenetic changes, which in a couple of generations stabilize cellular processes. Despite our increasing general knowledge of polyploidy and hybridization, detailed information on the links between such genomic responses to allopolyploidization and mechanisms involved in shaping long-term adaptive capacities in natural allopolyploid populations still remains limited.
We have studied gene expression and genome-wide
DNA methylation alterations in three sibling allotetraploid lineages of Dactylorhiza , an evolutionarily
288 complex genus of temperate terrestrial orchids.
Dactylorhiza majalis s.str., D. traunsteineri and D. ebudensis have formed independently through recurrent but unidirectional hybridization between the widespread diploids D. fuchsii and D. incarnata at different times during the last part of the Quaternary, but they differ radically in range and ecological preferences. Analyses of cDNA-AFLPs show a significant increase in the range of gene expression in the allopolyploid lineages, demonstrating a higher potential for phenotypic plasticity compared with either parent. Moreover, allopolyploid individuals express significantly more gene variants
(including novel ones) than the parents, providing clear evidence for an increase in biological complexity.
Significantly more genetic mutations have accumulated in the older D. majalis compared with the younger D. traunsteineri since their respective formations, but the opposite trend is visible in DNA methylation data.
Multiple origins of each tetraploid taxon (except D. ebudensis ) contribute to differential patterns of gene expression with a geographic structure. However, several transcripts and epialleles are conserved within each allopolyploid taxon but differ between taxa, indicating that habitat preferences shape similar expression patterns in independently formed allopolyploids. Under the continuous influence of natural selection, genomic alteration is found to have a direct impact on ecology and distribution of the three allopolyploids. Water availability and temperature appear to be the relevant factors correlated with these altered patterns of expression. In addition to stabilizing the allopolyploid genome, genetic and epigenetic alterations are key determinants of the adaptive success of the new polyploid species, potentially triggering reproductive isolation between the resulting lineages.
A phylogeny of the monocots, based on plastid genomes and additional genes of the plastid and mitochondrial genomes
Barrett, C 1 , Davis, J 1
1 Cornell University, New York, USA
Several major features of higher-level monocot phylogenetic relationships have been resolved by previous analyses, often with strong support, including the resolution of a series of major clades now conventionally recognized at the ordinal level (e.g.,
Pandanales, Liliales, Asparagales, and Zingiberales), and relationships among the orders (e.g., Commelinales +
Zingiberales). However, some aspects of inter- and intraordinal relationships have remained substantially unresolved, such as those among major groups within the commelinids, and among families and groups of families within Poales sensu APG. The Monocot Tree of Life project was established to address these and other questions, and a major goal has been to develop and analyze a comprehensive matrix of morphological and molecular data (representing all three major genomes) for a broad sample of monocots. It is intended that the phylogenetic structure and patterns of character evolution

inferred from this matrix will establish a basis for focused studies of monocot groups by the broader community of systematists. A preliminary version of the planned monocot data set has been assembled, including exemplars of more than 400 genera, representing all monocot families, with complete plastid genome sequences for ca. one-fourth of these taxa, and multiple gene sequences for others, including rbc L and mat K from the plastid genome, and atp1 and nad5 from the mitochondrial genome. This matrix provides strong support for most conventionally recognized orders of monocots and for relationships among these orders, as well as for relationships among major groups within most orders. Among the less conventional relationships with substantial support by this data set are: (1) the placement of Piperales or Piperales + Ceratophyllum as sister of the monocots; (2) the placement of Liliales and
Asparagales within a monophyletic group that includes no other taxa; (3) the placement of Dasypogonaceae and
Arecaceae as sister groups, with this clade placed as the sister of another, which includes all other commelinids; and (4) a strongly supported pectinate structure within
Poales sensu APG, with Bromeliaceae, Typhaceae, and
Rapateaceae diverging in succession from a clade in which a group consisting of Cyperaceae, Juncaceae,
Eriocaulaceae, Xyridaceae, and allied families is sister of a clade that corresponds in membership to Poales sensu
Dahlgren et al.
Phylogenomic analysts of transcriptomes sampled across monocot orders
Leebens-Mack, J
Barker, M 3
Steele, R 6
Stevenson, D 7
T 9
1 , Wickett, N 2
, Gonzales, MB
, McKain, M 1
, Anè, C 9 , Davis, JI 10
4
, McCombie, WR 8
, Ayyampalayam, S 1
, Vision, TJ 5
, Duarte, J 2
, Zomlefer, W 1
, Gandolfo, MA
Pineyro, A
K 14
12 , Alvarez-Buylla, E
, Wong, GK-S 15 , dePamphilis, C 2
10
, McNeal, J 1
, Pires, JC 6
12 , Barrett, R
,
, Givnish,
, Chase, M
13
11 ,
, Thiele,
,
,
1 Dept of Plant Biology, University of Georgia, Athens,
USA; 2 Intercollege Graduate Degree Program in Plant
Biology and Institute of Molecular Evolutionary
Genetics, Huck Institutes of the Life Sciences,
Pennsylvania State University, USA; 3 Dept of Ecology and Evolutionary Biology, University of Arizona, Tucson,
USA; 4 Computational Biology Group, Texas Advanced
Computing Center, University of Texas, USA; 5 Dept of
Biology, The University of North Carolina, USA;
7
6
8
Dept of Biological Sciences, University of Missouri, Columbia,
USA; New York Botanical Garden, Bronx, USA;
Spring Harbor Laboratory, USA; 9
Cold
Dept of Botany,
University of Wisconsin, Madison, USA; 10 LH Bailey
Hortorium and Dept of Plant Biology, Cornell
University, Ithaca, USA; 11 Jodrell Laboratory, Royal
Botanic Gardens, Kew, UK; 12 Instituto de Ecología,
Universidad Nacional Autónoma de México, Mexico;
13 School of Plant Biology, The University of Western
14 Australia, Perth, Australia;
Herbarium, Perth, Australia; 15
Western Australian
Dept of Biological
Sciences, University of Alberta, Edmonton, Canada
Understanding of phylogentic relationships, and gene and genome evolution is advancing at an unprecedented pace with the advent of next generation sequencing.
Transcriptome sequencing is playing a pivotal role in this advancement. Transcriptome assemblies for over 50 species distributed across all monocot orders are being sorted into gene families and hundreds of gene trees are being estimated. A species phylogeny is being inferred from low-copy genes and gene tree reconciliations are being preformed for all well-sampled gene families. The reconciled gene trees reveal a dynamic history of gene and genome duplication. The implications of gene and genome duplication will be discussed within the context of cladogenesis and evolutionary innovations throughout monocot history.
Using combined evidence trees to elucidate morphological evolution in the monocots
Thadeo, M 1 , Stevenson, D 1
1 The New York Botanical Garden, USA
We have developed a morphological matrix with 230 characters with 309 apomorphic states for 300 monocot genera. This matrix has been combined with molecular data from the nuclear, mitochondrial, and chloroplast genomes. Some traits exhibit high levels of homoplasy.
The fruits in the monocot superorders are predominantly capsular and thus it has generally assumed that the presence of baccate fruits in various unrelated monocot orders and families is the result of convergent evolution and of no value in phylogenetic analyses. For example, berries have evolved in multiple families in both the
Asparagales and Commelinales in unrelated groups.
Homoplasy is an indicator of different histories of what appears phenetically to be the same thing. By examining the development of baccate fruits across the monocots we are able to demonstrate different developmental patterns with different tissues giving rise to the baccate portions of the fleshy fruits in different groups thus removing the homoplasy of simply using the term baccate or fleshy fruit. In an effort to expand the scope of this study, we have focused on other phenetic features such as fenestrated and reticulate venation, epigynous flowers, arils, etc. by using the same approach of reciprocal illumination from total evidence trees combined with comparative developmental studies across the monocots.
This leads to new character descriptions by comparing, for example, development of flowers and fruits from select species with baccate fruit types in the monocots and understanding that they come from the independent evolution of berries derived from the three tissues of the pericarp; the exocarp, the mesocarp, and the endocarp and varying combinations of the three. Thus, the new descriptors representing different developmental pathways become non-homoplasious synapomorphies while simultaneously contributing a better understanding of the systematics and biology of the monocots.
An update of fossil data on the evolution of monocots, with emphasis on the Southern Hemisphere
Conran, JG 1 , Bannister, JM 2 , Lee, DE 3
1 Australian Centre for Evolutionary Biology and
Biodiversity, School of Earth and Environmental
3
Sciences, University of Adelaide, Australia; 2 Dept of
Botany, University of Otago,Dunedin, New Zealand;
Dept of Geology, University of Otago, New Zealand
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The record of fossil monocotyledons is updated based on recent finds globally, covering diverse groups including the Alismatales, Araceae, Arecaceae, Cyclanthaceae and bambusoid Poaceae, in some instances dating back to the
Cretaceous. In particular, reports for mummified leaf fossils with good cuticular preservation from the
Southern Hemisphere are summarised. A number of families in several different orders show significant fossil age or range extensions, such as Ripogonaceae in the
Eocene of Tasmania and New Zealand and calamoid palms from the Eocene of southern New Zealand. There are also reliable first macrofossil records for several families or subfamilies including Orchidaceae:
Epidendroideae ( Dendrobium and Earina ), Asteliaceae
( Astelia ), Asparagaceae: Hemerocallidoideae
( Dianella / Phormium ) and Alstroemeriaceae:
Luzuriagoideae ( Luzuriaga ) from the Miocene of New
Zealand. The biogeographic and palaeoecological implications of these records, especially for tropical or subtropical taxa at occurring at mid to high southern latitudes as well as the use of these records for improved dating of monocot divergence ages are also discussed.
Monocot phylogenetics: from mutually incongruent hypotheses to a consensus based on molecules
Chase, M 1
1 Royal Botanic Gardens Kew, UK
Before the advent of molecular phylogenetics, there were several competing hypotheses for how evolution proceeded in the monocot lineage, some expressed in phylogenetic terms (which families were primitive versus advanced) and others in the context of primitive versus advanced monocot characters. Most were not particularly clear about branching order, except for Dahlgren and coworkers, but they too sometimes confused taxa with characters in their discussion. Once we entered the molecular era, many of the previous ideas have been forgotten, so it is worthwhile discussing them from the perspective of the molecular results to see to what extent the ideas of primitive versus advanced characters still apply. Fortunately, not a lot has changed on the molecular side from the first studies with just plastid rbc L, but there are still a few areas where further effort is required (e.g. the base of the commelinid clade and some relationships in Poales). A look to the what the future may hold is focused on what the era of whole-genome sequencing may be able to tell us.
Early monocot evolution and monocot ecology
Coiffard, C 1 , Mohr, BAR 1 , Bernardes-De-Oliviera, MA 2
1 Museum fur Narturkunde, IBerlin, Germany; 2 Institute of Geosciences, University of São Paulo, Brazil
The lower Cretaceous angiosperm macrofossil record already shows the three major groups of angiosperms that we know today, i.e. ANITA ( Pluricarpellatia ,
Scutifolium ), Magnoliids (e.g. Archaeanthus ,
Endressinia , Araripia ) and Eudicots ( Sapindopsis ).
However, monocots with a high diversity today – they constitute about one-fourth of living angiosperms – are known only by pollen grains, charcoalified flowers and a few leaves, such as possibly Acaciaephyllum . The fossil
290 record remains scarce up to at least the Turonian with the appearance of Arecaceae and Pandanaceae. The occurrence of two anatomically preserved taxa in the late
Aptian Crato Formation of northeastern Brazil shows that monocots had already started to diversify during the
Lower Cretaceous.The first taxon consists of isolated leaves that have a remarkable affinity to recent basal
Araceae. The second taxon is preserved as a whole plant, including roots, slender parallel veined leaves and flowers and shows affinities to core monocots. Both taxa seem to have grown in differing habitats under very different ecological conditions. Thus the scarceness in the fossil record does not represent a delayed diversification but most likely corresponds to a taphonomic bias.
Araceae in space and time: a dated tree for representatives of 114 genera based on chloroplast and nuclear DNA
Nauheimer, L 1 , Cusimano, N 1 , Chartier, M 2
1
Renner, S 1
, Bogner, J 2 ,
University of Munich, Germany;
Toulouse, France
2 University of
The Araceae family is among the oldest monocot lineages, as implied by their position in the angiosperm tree, as well as macrofossils from the Cretaceous that have been assigned to particular subfamilies of Araceae, such as Orontium mackii, Lasioideaecidites bogneri , and
Albertarum pueri from the late Cretaceous, and
Caladiosoma messelense, Petrocardium wayuurum and
Aracephyllites tertiarus from the Middle Eocene.
Extremely old pollen from the Early Cretaceous (Late
Aptian), described as Mayoa portugallica , also represents
Araceae. We have generated a fossil-calibrated dated phylogeny based on an almost complete genus sampling
(114/118 genera included) and a dataset of 5000+ chloroplast and nuclear nucleotides. Today, Araceae comprise c. 4000 species, most of them in the Neotropics and SE Asia, and molecular data indicate that the family is sister to the remaining Alismatales. Formal biogeographic analyzes imply early Araceae diversification in Laurasia, but outgroup coding for ancestral area reconstruction is problematic or dubious.
The talk will present diverging ages of key nodes and where possible relate them to paleoclimate, continental plate movement, and long distance dispersal. A second focus is the controversial assignment of fossils to clades due to the convergence of aroid leaf venation patterns. To address this problem, we performed cross validation of fossil placement to assess the influence of each fossil on the inferred dates individually. The ages of the deepest nodes depend almost entirely on the maximum age permitted for the root. Without a Late Aptian root age constraint, the stem age of Araceae exceeds that generally accepted for the angiosperms. However, as argued by S. Smith et al. (2010) angiosperms could well be older than is apparent from their fossil record.
New insights into the biogeography of the Austral floristic realm from a complete phylogeny for the
Alstroemeriaceae
Chacón, J
Renner, SS 1
1 , Camargo De Assis, M 2 , Meerow, AW 3 ,

1 University of Munich, Germany; 2
Ambiente, Jaguariúna, São Paulo, Brazil;
SHRS, National Germplasm Repository
EMBRAPA Meio
3 USDA-ARS-
Alstroemeriaceae consists of the Neotropical
Alstroemerioideae with the genera Alstroemeria (75 spp.) and Bomarea (120 spp., mostly in the Northern Andes except for four species in Chile) and the disjunct
Luzuriagoideae with the genera Luzuriaga (4 spp.) and
Drymophila (2 spp.). Luzuriaga has three species in temperate South America and one, Luzuriaga parviflora , in New Zealand, while the two species of Drymophila occur in Eastern Australia and Tasmania. This makes the
Alstroemeriaceae a typical member of the Austral floristic realm. We have sequenced 138 species from throughout the family's distribution range, using
Colchicaceae as outgroup. A total of 3189 aligned nucleotides from the chloroplast DNA regions mat K, ndh F and rbc L, the mitochondrial gene matR, and the nuclear ribosomal ITS were analyzed. A recently discovered fossil of Luzuriaga from New Zealand (to be described J. Conran and colleagues) was used to constrain relaxed and strict molecular clock models, alternatively with fossils of Smilax and the root age of
Liliales. Maximum likelihood trees show that
Alstroemeria and Bomarea are mutually monophyletic and together form the sister clade of Drymophila and
Luzuriaga . The Brazilian species of Alstroemeria form a clade that is embedded within the Chilean and
Argentinean species. Luzuriaga parviflora is embedded among the South American luzuriagas. Based on the clock models, Alstroemeriaceae originated in Cretaceous
East Gondwana, coeval with other members of the
Austral floristic realm, such as Proteaceae,
Atherospermataceae, Restionaceae, and Nothofagaceae.
The diversification of the Alstroemerioideae and
Luzuriagoideae clades occurred during the Eocene, c. 44
Ma ago, when Australia and South America where still partly connected, and Antarctica was still forested. In southern South America, Alstroemeria began to diversify c. 26 Ma ago, before the main uplift of the Andean cordillera and it then expanded east into Brazil, where it radiated during the middle Miocene, c. 13 Ma ago.
Bomarea diversification commenced later, c. 18 Ma ago, and this lineage expanded and diversified northwards throughout the Pliocene and into the Pleistocene, as cloud forests and páramos began covering the cordilleras.
The presence of L. parviflora in New Zealand is likely a result of long distance dispersal from Patagonia, and the
New Zealand Luzuriaga fossil appears to represent a stem lineage.
Plastome-scale phylogenetics and the evolution of
Poaceae
Duvall, MR 1 , Columbus, JT 1 , Ingram, A 1
1 Northern Illinois University, USA
, Jones, SS 1
Poaceae are a diverse and species-rich evolutionary radiation with a complex history. Systematic studies of the family can benefit from the phylogenetic information content of whole plastomes. Complete plastomes or draft plastomes composed of the conserved protein coding loci were analyzed for species in 25 genera, seven of which were not previously published, including a recently completed plastome from Pharoideae. These species represent the breadth of ecological diversity in the family including temperate and tropical species, widespread adventives and narrow endemics, woody perennials and herbaceous annuals. Biogeographic diversity was also well represented with species from North and South
America, Mexico, Eurasia, Africa, China, and Australia.
Phylogenetic representation included species from three early-diverging lineages and from six subfamilies in the
PACMAD and BEP clades. Plastomes from other graminoid Poales, Ecdeiocoleaceae, Joinvilleaceae,
Flagellariaceae, and Restionaceae, which were determined by the monocot tree-of-life consortium, were included as outgroups. The 228 priming sites used in amplification and sequencing were over 75% conserved within Poaceae, but less than 50% conserved in other
Poales tested ( Joinvillea , Ecdeiocolea , and
Lepidosperma ). Over 7,000 parsimony-informative characters supported plastome-scale phylogenies and maximum or near-maximum bootstrap support was observed in all analyses. Microstructural characters, such as introns, large insertions, and pseudogenes, which are thought to be grassspecific, were surveyed.
Anomochlooideae, Puelioideae, and Pharoideae showed the expected order of topological divergence, and
Anomochlooideae was monophyletic (two species).
However, Anomochlooideae shared fewer synapomorphic microstructural characters with other
Poaceae than Puelioideae and Pharoideae. The unusual inflorescence morphologies of Anomochlooideae assume somewhat greater systematic significance when viewed in the context of their distinctive plastomes. Pharus divides a long internal branch in the backbone of the grass subtree, leaving only one long internal branch in
Poaceae – that subtending the PACMAD plus BEP clades. This branch likely represents either accelerated diversification of this large group or extinctions along the ancestral lineage to this clade. Our results are in agreement with a broadly delimited Andropogoneae, and do not support the monophyly of Maydeae. Concurrent with the highly conserved nature of the coding regions of the plastome was considerable variation in noncoding introns and spacers useful for studies at lower taxonomic levels. Readily interpreted mutational patterns were observed, such as small inversions of the loop in hairpinloop regions and indels resulting from slipped-strand mispairings. Events altering gene expression, the locations of the boundaries of the inverted repeats, and the overall sizes of plastome regions were also observed.
The rich accumulation of mutations in the complete plastomes of Poaceae is indicative of the utility of these small genomes for extensive future phylogenetic and systematic studies.
Lycium
(Solanaceae): diversity, dispersal, and dating
Levin, R 1 , Miller, J 1
1 Amherst College, MA, USA
Lycium (Solanaceae) is one of the largest (ca. 88 species) and most geographically diverse genera in Solanaceae, being one of only three genera in Solanaceae with a
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cosmopolitan distribution. The genus is most species-rich in southern South America, with additional hotspots of species diversity in southwestern North America and southern Africa. This extensive distribution is in contrast to the restricted South American distribution of two small genera ( Grabowskia and Phrodus ) that, together with Lycium , comprise tribe Lycieae. Furthermore, the closest relatives of Lycieae, including the species-rich genus Nolana , are all restricted to South America. The genus Lycium is unique among these taxa in exhibiting variation in mating system, with both dimorphic and hermaphroditic species, as well as associated differences in ploidy. Using a combination of low-copy nuclear regions (GBSSI, several COSII markers) and plastid sequence data from the vast majority of species, we infer evolutionary relationships across Lycium and among the three genera within the tribe, employing analysis methods that allow for the simultaneous inference of species trees from multi-locus data. Using this understanding of species relationships, we examine biogeographic patterns, including an estimate of dispersal dates inferred using BEAST. Tribe Lycieae appears quite young, arising in South America ca. 5 Ma, with dispersal from the New to the Old World occurring ca. 3.5 Ma. We also re-evaluate generic circumscriptions within tribe
Lycieae, including discussion of fruit diversity within the tribe. Fruit morphology has traditionally been used to differentiate the three genera of Lycieae, and we will explore this fruit diversity within the context of an updated classification that resolves the paraphyly of
Lycium .
Prickly phylogenies: unraveling relationships in
Solanum
subgenus
Leptostemonum
Stern, S 1 , Bohs, L 1
1 University of Utah, USA
The genus Solanum (Solanaceae) is one of the largest and most economically important plant genera. One of the largest groups within the genus is the Leptostemonum clade with ca. 350–450 species. This clade is also known as the 'spiny solanums' due to the presence of epidermal prickles, and it basically conforms to Solanum subgenus
Leptostemonum of traditional classifications. Here we present a phylogeny of the spiny solanums with a goal of examining the monophyly of sections recognized within this large clade. Of particular interest is the Micracantha clade, a group of ~12 species that has challenged taxonomists. We clarify the species composition of this clade and further investigate specific relationships within the group using DNA sequence data from eight markers.
We examine the utility of these newly developed markers for inferring relationships within the Micracantha clade.
These efforts help to define monophyletic groups within the large Leptostemonum clade and will aid in taxonomic revisions of groups within Solanum subgenus
Leptostemonum .
Comparative gene function in the development of fleshy and dry fruits in Solanaceae
Litt, A 1 , Meyer, R
Mohamed, A 1
1 , Pabon-Mora, N 1 , Nowogrodzki, A 1 ,
1 The New York Botanical Garden, USA
292
The Solanaceae are characterized by a diversity of fruit types, but most species produce either a dry capsule (e.g.
Nicotiana sylvestris ) or a berry (e.g. Solanum lycopersicum ). Phylogenetic reconstruction suggests that the capsule is the ancestral fruit type in the family; concurrent with the origin of the species-rich
Solanoideae clade there was a dramatic shift to berry formation. The goal of this project is to identify some of the key genetic differences that determine whether a
Solanaceae fruit will be dry or fleshy. Solanaceae are an ideal system in which to address such developmental questions: there is significant morphological variability but considerable sequence similarity among species, and they are amenable to both VIGS and stable genetic transformation, making them excellent subjects for the study of differential gene function. Using standard histological techniques and in situ hybridization, we have previously identified four comparable landmark stages in the development of the fruit of S. lycopersicum and N. sylvestris . Identification of these stages provides a foundation for comparative transcriptome analyses, with the aim of identifying genes that show differential activity during the development of the two fruit types, and that may play a role in determining the specific traits that characterize dry and fleshy fruits. We have implemented both microarray and next-generation sequencing strategies to identify genes that are expressed differentially both between the two species and across the four developmental stages. Our preliminary analysis of genes that are upregulated in each species at the time of fertilization and the concurrent onset of rapid cell division (Stage 2) identified several transcription factors, signaling factors, and genes involved in relevant processes such as cell wall expansion or lignification as being differentially regulated. We selected 12 genes for more in-depth expression analyses. RT-PCR analysis of expression in vegetative and reproductive tissues suggests that some of these genes are uniformly expressed in all tissues, with no fruit-specific activity.
Other genes show relatively increased or decreased expression during one or more of the four stages of fruit development suggesting a specific role in fruit development. Quantitative real-time expression analysis of seven of the genes at the four different fruit developmental stages confirms these patterns and highlights some notable trends such as significant differences in the expression of candidate genes in the berry and capsule at stage 3, or several genes that appear to be expressed constitutively in one fruit type but that vary developmentally in the other. These are candidates for functional analysis to determine their specific role in fruit development. RNAi analysis of one candidate gene encoding an ethylene receptor, already known to repress tomato ripening, showed that it is required for proper drying of the N. sylvestris capsule prior to dehiscence.
We are currently using VIGS to evaluate the function of additional genes in both species.
Genome downsizing in allopolyploid
N. tabacum
: how does next generation sequencing depth influence
1 results?
Renny-Byfield, S
Nichols, R 1
1 ,
, Macas, J 4
Kovarik, A 2 , Grandbastien, M-A 3
, Novák, P 4 , Leitch, AR 1
,
Queen Mary University of London, School of Biological and Chemical Sciences, UK; 2 Institute of Biophysics,

Academy of Sciences of The Czech Republic;
Jean-Pierre Bourgin, INRA-Versailles, France;
3
4
Institute
Biology
Centre ASCR, Institute of Plant Molecular Biology,
Czech Republic
We used next generation sequencing to characterize and compare the genomes of the recently derived allotetraploid, Nicotiana tabacum (< 200,000 years old), with its diploid progenitors, N. sylvestris (maternal, Sgenome donor) and N. tomentosiformis (paternal, Tgenome donor) using data from 454 pyrosequencing and
Illumina genome analyser. We have previously shown using 454 technology that all major types of retroelements found in angiosperms are underrepresented in tobacco, especially from the paternally derived Tgenome. These data are reviewed and we show here how our understanding of the genome of these three species are influenced by greater read depths but shorter read lengths generated by Illumina technology
The potato genome and its comparison with other plant genomes
Yang, B 1 , Xu, X 1 , Zhang, Z
1 1BGI Shenzhen, China; 2
2 , Li, R 1 , Qu, D 2 , Huang, S 2
Institute of Vegetables and
Flowers, Chinese Academy of Agricultural Sciences,
China
Potato ( Solanum tuberosum ) is the third most important food crop and an increasingly important alternative to the cereals. Potato feeds the world population with its tubers that function in nutrient storage and asexual reproduction. However, we still know little about the biology of the potato tuber, and the autotetraploid and highly heterozygous genome of the plant makes it recalcitrant to modern molecular breeding. Here we report the 850 Mbp genome sequence of potato obtained from both a doubled monoploid clone. The potato genome sequence is the first of an Asterid plant, a major phylogenetic clade that includes ~1/3 of all flowering plant species. It significantly differs from the eight previously sequenced genomes of Rosid plants with
~15,000 lineage-specific genes grouped in about ~5,500 gene families. Further comparison with genome and EST sequences from other Solanaceae species and compositae species delimited ~3,500 genes in 2,800 gene family to be putative Asterid specific. Expression profiling and comparative analyses to non-tuber bearing relatives provided novel insights on tuber biology, including tuber development, starch biosynthesis, nitrogen storage, and immunity. The genome sequence provides a new platform for the genetic improvement of this crop with impact to global food security.
Clocks, clades and continents: evaluating hypotheses of vicariance, dispersal, and time in Southern
Hemisphere Myrtales (Combretaceae, Myrtaceae,
Metrosideros
)
Sytsma, K 1 , Berger, BA
1 Dept of Botany, University of Wisconsin–Madison, USA
The Myrtales represents a clade of widely-dispersed as well as endemic families and thus raises intriguing biogeographic questions related to both the processes and timing of the disjunctions seen at continental and oceanic island scales. Our previous studies have examined issues of vicariance and dispersal in both Myrtaceae (and associated families) and the disjunct genus Fuchsia
(Onagraceae) – both largely restricted to the Southern
Hemisphere – using fossil calibrated DNA phylogenies.
We expand upon these studies by examining the historical biogeography of the entire order Myrtales with the inclusion of many taxa of the pantropical
Combretaceae and additional species of Metrosideros
(Myrtaceae). Molecular phylogenetic hypotheses of
Myrtales, Combretaceae, and Myrtaceae are presented that include considerably greater taxa sampling and additional genomes and gene regions. We use BEAST,
Lagrange, and r8s analyses with many more fossils to place time into the biogeographical analyses of these lineages. The Combretaceae has been a largely missing entity in past studies of the Myrtales. With its distinctive disjunct distribution across the Southern Hemisphere, the family is critical for full understanding of the evolution of the Myrtales in time and space and for clarification of the role of Gondwanan breakup in the order and in
Combretaceae. The phylogenetic and biogeographic analyses place the Combretaceae stem radiation back into the Cretaceous (ca. 90mya) with a subsequent, but old divergence of the enigmatic, African Strephonema . The crown radiation of the remainder of Combretaceae
( Combretum , Terminalia , and mangrove subclades) is
Tertiary. We show that dispersal hypotheses are supported for the spread of Combretum and Terminalia in Austral–Pacific. In addition, we expand upon the sampling of Metrosideros and allies (especially Austral-
Pacific species) to test alternative and competing hypotheses of the timing of the dispersal of this clade across Austral–Pacific island chains. This analysis of
Metrosideros and allies was performed by placing this lineage, for the first time, in the context of our larger historical biogeographic analysis of Myrtaceae. The timing of the dispersal of this lineage across the Pacific and into Hawaii is compared and contrasted to previous and widely differing hypotheses of timing that did not use explicit fossil calibration as here presented.
Austral origins and Pacific radiations: the historical biogeography of
Astelia
s.l. (Asteliaceae)
1
Birch, J 1 , Morden, C 2 , Keeley, S 1
University of Hawaii at Manoa, Botany Dept, Honolulu,
USA; 2 University of Hawaii at Manoa, Pacific Cooperative Studies Unit, USA
The flowering plant family Asteliaceae (Asparagales) contains 4 genera and approximately 36 species with a center of diversity in New Zealand. Previous studies have dated the Asteliaceae stem lineage to the late Cretaceous
(79–104 my), that is, during the period of separation of
New Zealand from East Gondwana. While Astelia s.l. is found on three Gondwanan landmasses (Australia, New
Zealand, and South America), it is also well represented in much younger areas, for example, on oceanic islands with recent geological origins such as the Marquesan (5.5
293

my) and Society (4.5 my) Islands. The phylogenetic reconstruction of Astelia s.l. and the Asteliaceae, based on a combination of chloroplast ( trn L, ps bAtrn H, rps 16, and pet Lpsb E) and nuclear (NIA-i3) sequence data, is presented and used to examine the biogeographic history of the genus. Relaxed molecular clock analyses date the crown radiation of Astelia s.l. to ca. 46 my (95% HPD:
32–52 my), followed by radiation of the subgenera during the early Miocene and late Oligocene (12–28 my).
These dates are long after the separation of the
Gondwanan landmass into separate continents; therefore, the current distribution of Astelia s.l. is best explained by trans-oceanic long-distance dispersal rather than vicariance. Results suggest two dispersal events between
New Zealand and Australia and a single dispersal between New Zealand and South America. Neither
Eastern nor Western Pacific Island Astelia s.l. taxa are monophyletic. At least four direct, long distance dispersal events are required to explain the distributions of Astelia s.l. taxa on island archipelagos. A steppingstone dispersal pathway is proposed for Astelia s.l. from
New Zealand to the Austral Islands and from there to
Hawaii. In addition, nesting of the Marquesan species in an otherwise Hawaiian clade suggests a secondary dispersal for Astelia s.l. from Hawaii to the Marquesan
Islands. Analyses are underway to identify morphological and ecological traits that are correlated with long-distance dispersal in Astelia s.l. Morphological and ecological features including fruit type, fruit and seed size, and association with wetland habitats are under investigation to assess their evolutionary origin/s and to determine their selective advantage for long-distance dispersal in the Astelia s.l. radiation.
Evolution and diversification of the Austral–Pacific family Goodeniaceae
Jabaily, R 1 , Motley, T 1 , Gustafsson, M 2 , Howarth, D 3
1 Old Dominion University, Virginia, USA;
University, Denmark; 3
2 Aarhus
St Johns University, Queens, New
York, USA
The Goodeniaceae, with over 400 species in eleven genera, has radiated extensively in Australia, particularly in the Southwest Botanical Province and throughout the
Pacific Islands, including Hawaii and New Caledonia.
Taxa exhibit marked variability in floral color and symmetry, seed, pollen and indument morphology, and type of pollinator guides. Goodeniaceae is also of systematic interest because of its position within the
Asterales, sister to Asteraceae and Calyceraceae. A comprehensive molecular systematic study of the family with ample sampling of all recognized genera is presented in order to test the current classification scheme, as well as hypotheses of morphological character evolution and biogeographic patterns throughout the Austral–Pacific basin. The cpDNA regions trn L-F and mat K were sequenced and provided ample variation to resolve all intergeneric and most interspecific relationships with high support. Two major clades of Goodeniaceae are recovered – the
Dampiera / Lechenaultia / Anthotium group, all with connate anthers, is sister to a larger clade with primarily adnate anthers composed of
Goodenia / Scaevola / Coopernookia and several other monotypic genera, including Brunonia , formerly placed
294 in the monotypic Brunoniaceae. All genera are monophyletic with the exception of Goodenia , which contains Verreauxia , Velleia and several monotypic genera. Subgeneric taxonomy generally does not resolve as monophyletic. Molecular dating across the family finds that deep phylogenetic divisions predate the break of Australia from Antarctica, with more recent radiations in Goodenia and Scaevola corresponding to aridification and climate change events of the Pleistocene. Repeated dispersals out of Australia and into the Pacific Island archipelagos are recovered, aided by unique fruit characteristics. The Australian Southwest Botanical
Province was the ancestral area for one of the major clades, with movements between the major Australian biomes happening frequently during the evolution of
Goodeniaceae.
Dragons in the mist: origin, dispersal and diversification of the Australasian genus
Dracophyllum
(Richeeae, Ericaceae)
Wagstaff, SJ
Crayn, DM 4
1 , Dawson, MI
, Steane, DA 5
1 , Venter, S 2
, Lemson, KL 6
, Munzinger, J 3 ,
2
1 Allan Herbarium, Landcare Research, New Zealand;
Botanical and Environmental Consultant, Trinity Beach,
Queensland, Australia; 3 IRD, UMR AMAP, Laboratoire
5 de Botanique et d’Ecologie Végétale Appliquées,
Herbarium NOU, New Caledonia; IRD, UMR AMAP,
Montpellier, France; 4 Australian Tropical Herbarium,
James Cook University Cairns Campus, Australia;
School of Plant Science and CRC Forestry, University of Tasmania, Hobart, Australia; 6 Centre for Ecosystem
Management & School of Natural Sciences, Edith Cowan
University, Western Australia
The genus Dracophyllum (Ericaceae) has a fragmented distribution across Australasia, but reaches the greatest level of species richness and morphological diversity in
New Caledonia and New Zealand. About 50 species are currently recognized, varying from cushion plants to trees up to 14 m tall. They are conspicuous in montane rainforests and wet alpine heathland and are commonly known as ‘dragon heath’ because of their distinctive growth form. We investigated evolutionary processes that contribute to this disparity in species richness. We compiled two complementary data sets for the chloroplast-encoded genes mat K and rbc L that included representatives of Dracophyllum , its close relatives
Richea and Sphenotoma (together constituting Tribe
Richeeae), along with more distant relatives in the
Ericaceae. The sequences were subjected to Bayesian and maximum likelihood analyses to assess the robustness of our phylogenetic inferences. We chose four calibration points based on the fossil record and geological events, but nonetheless there was considerable uncertainty surrounding our divergence estimates. In our analyses,
Tribe Richeeae formed a monophyletic group that diverged during the Eocene (at least 33.3 Ma) with a crown radiation during the early Miocene (at least 16.5
Ma) that resulted in two disjunct lineages. The crown radiation coincoides roughly with the onset of aridification in central Australia and fragmentation of the rainforests in eastern Australia. The Western Australian genus Sphenotoma formed an isolated evolutionary lineage, while Dracophyllum and Richea together formed a second lineage that was widely distributed in Eastern

Australia, Lord Howe Island, New Caledonia and New
Zealand. The relationships of the Tasmanian endemic,
Dracophyllum milliganii , remain an enigma. It was ambiguously placed in our analyses, emerging as sister either to Sphenotoma or to the Dracophyllum / Richea clade. We documented two instances of long-distance dispersal to New Caledonia and New Zealand in the late
Miocene to early Pliocene. However the recent discovery of Oligocene–early Miocene macrofossils that resemble
Dracophyllum predates these molecular estimates by several million years. The low level of sequence divergence suggests a rapid and recent species radiation in these two island archipelagos largely within the last 3–
6 million years. This radiation may have been spurred by the Pliocene uplift of the Southern Alps in New Zealand and by episodes of glaciation during the Pleistocene..
Biogeography and character evolution in the
Loganieae, Loganiaceae on continental islands and oceanic archipelagos
Motley, T 1
1 Old Dominion University, Virginia, USA
The subfamily Loganieae is a mostly woody lineage that has its center of origin in Australia and the genera have dispersed to and diversified in the islands of the Pacific.
Labordia is a Hawaiian endemic genus and Geniostoma is distributed throughout the other islands of the Pacific and Indian Oceans. These two sister genera are most closely related to the Australian-centered genera Logania and Mitrasacme. The question exists as to whether
Labordia and Geniostoma should be treated as a single genus or as two separate genera. These genera are separated primarily by the terminal inflorescences with of more showy corollas in the former genus and axillary inflorescences in the latter. The reason this debate has remained unsettled is the presence of Geniostoma rapense with terminal inflorescences located on the isolated island of Rapa iti. This species has been suspected to be the 'missing link' necessary for understanding the generic relationships. Recent field research on Rapa has provided material for this study.
Phylogenetic analyses suggest that Labordia and
Geniostoma should be treated as distinct genera.
Labordia , consists of two morphologically distinct sections, which have arisen from a single colonization event. Dioecy has evolved several times in the
Loganieae. Furthermore, the genus Logania is not monophyletic. Additionally, in order to gain better phylogenetic resolution among the quickly evolving species of Labordia AFLP fingerprinting markers have been used in association with sequence data. These markers are congruent with the sequence data and in a combined analysis provide more resolution and biogeographic signal in the Hawaiian radiation.
Biogeography of South American basal Asteraceae
Katinas, L 1 , Crisci, JV 1
1 División Plantas Vasculares, Museo de La Plata,
Argentina
The Asteraceae or Compositae, with ca. 1700 genera and
25,000 species, is the most diverse family of flowering plants in terms of numbers of genera and species and has a cosmopolitan distribution found in every continent except Antarctica. The key morphological feature for the family is the capitulum, or tight cluster of sessile flowers on a receptacle, which constitutes an efficient reproductive unit that may have evolved to serve as a larger pollinator target. The most recent phylogenetic trees of Asteraceae place groups Barnadesioideae and
Mutisioideae sensu lato as a basal grade within the family. These groups are either endemic to or mainly distributed in South America, with high taxa concentration in Patagonia and the Andean ranges. Fossil evidence and current distribution of basal members of
Asteraceae and closely related families suggest an origin of Asteraceae linked to Late Gondwanan events, with first Patagonian (South America) fossils unequivocally assignable to the Middle Eocene (47.5 million years ago).
This was followed by an explosive radiation of the lineages in southern South America in areas subject to the Andean cordilleran orogeny and Quaternary glaciations events, followed by a quick colonization of other areas. Fossil evidence together with extant taxa distribution are two types of evidence that help to reconstruct the historical biogeography of the family. The latest fossil findings of basal groups in southern areas together with historical biogeographic studies of extant taxa are presented to help explain the distributional patterns of members of the family in South America.
Evolutionary history of Arecaceae tribe Cocoseae inferred from seven WRKY transcription factor loci
Meerow, A 1 , Noblick, L 2 , Francisco-Ortega, J 3 ,
2
1
Nakamura, K 1
USDA-ARS National Germplasm
Montgomery Botanical Center, USA;
International University, USA; 4
Repository;
3 Florida
Fairchild Tropical
Botanical Garden, Florida, USA
The Cocoseae is one of 13 tribes of Arecaceae subfam.
Arecoideae, and contains a number of palms with significant economic importance, including the monotypic and pantropical Cocos nucifera , the coconut, and African oil palm ( Elaeis guineensis ). Using seven single copy WRKY transcription factor genes, we previously inferred a well resolved phylogeny and chronology of the subtribe Attaleinae. In that study, we resolved a monophyletic relict African clade as sister to the American genera, including Cocos . We determined that Lytocaryum is best treated as part of Syagrus , resolved Syagrus sensu lato as sister to Cocos , and presented a novel clade uniting Parajubaea with Butia and Jubaea that is also supported by leaf anatomical data.
Branch nodes were congruent with major paleogeographic events. We now extend our sequence analysis across the entire tribe, with Roystonea and
Reinhardtia as outgroups. Three subtribes, Attaleinae,
Bactridinae and Elaeidinae, are consistantly resolved with strong support. The monotypic Amazonian Barcella is sister to Elaeis . The genus Hexopetion is well supported as distinct from Astrocaryum . The inclusion of
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Gastrococos in Acrocomia is also supported. Two methods of species tree generation will be compared: gene tree parsimony with DupTree, and Bayesian approximation with BEST. A scenario for the overall biogeograpic and evolutionary history of the tribe will be presented.
Plastid genomes and palm phylogenetics
Davis, J 1 , Leebens-Mack, J
Stevenson, D 3 , Zomlefer, W 2
2 , Barrett, C 1 , Comer, J 2 ,
1 Cornell University, New York, USA;
Georgia, USA;
2 University of
3 New York Botanical Garden, USA
The palm family (Arecaceae or Palmae) has been the subject of intensive phylogenetic analysis, using molecular and morphological characters. Previous studies indicate that genes of the plastid genome, and the genome structure itself, evolve extremely slowly in comparison with other plant groups. A well supported phylogenetic structure for the family has been developed, and current understandings of relationships within the family have been incorporated in a modern systematic treatment. However, several key groups within the family have weak support, and although there is strong support for the placement of palms within the commelinids, their precise placement among other commelinid families remains questionable. The Monocot Tree of Life project was established to address several matters relating to the overall phylogenetic structure of monocots, and the palms are a major point of focus for these studies.
Complete plastid genome sequences have emerged as an important tool for phylogenetic studies, and sequences for major plant lineages have begun to accumulate rapidly. Plastid genome sequences can be generated by traditional and 'new technology' sequencing methods.
Using the latter approach, we have assembled a set of more than 20 plastid genomes for representative palms from all subfamilies and more than half of the recognized tribes. Preliminary analyses of these and additional plastid genomes representing other monocot groups place the Arecaceae as sister of Dasypogonaceae, and place this two-family clade as sister of a clade that includes all other commelinid monocots (i.e., Commelinales,
Zingiberales, and Poales sensu APG). A detailed analysis of nucleotide sequence variation in coding and noncoding regions of the plastid genome corroborates previous observations that the plastid genomes of palms evolve quite slowly. However, structural mutations do mark several clades within the palms, as well as the grouping of palms with Dasypogonaceae. Continued integration of whole plastome sequences into phylogenetic matrices consisting of smaller numbers of genes sampled for larger numbers of taxa is yielding highly structured phylogenies with strong support for critical nodes.
Palm supertrees – worthwhile expedient or sloppy short-cut?
Baker, WJ 1
1 Royal Botanic Gardens, Kew, UK
A recently published supertree study has provided the first complete genus-level phylogeny of palms
296
(Arecaceae/Palmae). The palm supertree topology is highly resolved with widespread support. As a result, it has been used by the broader community of palm researchers for a variety of purposes such as the construction of a new phylogenetic classification of the family, comparative analyses of reproductive biology and diversification analyses. Supertree methods offer an expedient for synthesising disparate data sources into often very large trees while minimising the detrimental impact of missing data. However, detractors of supertrees argue that they are dislocated from 'real' data and are thus less valid phylogenetic hypotheses than those inferred by conventional means. Other negative features of supertrees have also been highlighted, such as nonindependence among input trees, a lack of signal enhancement and the recovery of relationships that are not present in any of the input trees. The palm supertree has influenced recent research on the family, but should this phylogenetic hypothesis really be trusted? Do these methods have a place among the growing body of more conventional phylogenetic analyses and new genomescale approaches? Do the benefits of the alternatives outweigh the efficiency of supertrees? In this talk, these issues will be explored with a view to determining the future role of supertrees in the palm phylogenetic agenda and their likely contribution to comparative biology research on the family.
Diversification of palms in the earliest tropical rain forests
Couvreur, T 1 , Forest, F 2 , Baker, W 2
1 Institut de Recherche pour le Développement (IRD),
France; 2 Royal Botanic Gardens, Kew, UK
More than half of the world’s biodiversity is contained within tropical rain forests (TRF), yet the origin and diversification of this diversity remains largely unexplained. Major hypotheses of TRF diversification include gradual accumulation of diversity over long periods of time or rapid speciation events either ancient or recent. Here we present the first investigation of the mode and tempo of TRF diversification based on a complete genus-level sampling for an important TRFrestricted plant family. Palms (Arecaceae/Palmae) are one of the most ecologically important and characteristic components of TRF worldwide and represent a model group for the investigation of TRF evolution over long periods of time. We estimated speciation events under a
Bayesian framework using a relaxed clock approach, while spatial origins of the family were inferred under a maximum likelihood method that implements the dispersal–extinction–cladogenesis model. An analysis of diversification was carried out based on lineage-throughtime plots (LTT) plots. We provide evidence that diversification of extant lineages of palms started during the mid-Cretaceous about 100 million years ago (Ma) in
Laurasia. This correlates with the first fossil indications interpreted as TRF in North America during the mid-
Cretaceous as well as the diversification of angiosperms in general. Our results suggest that palms played a major role in TRF diversification since the origin of the biome.
Palms conform to a constant diversification model of
TRF evolution (the 'museum' model), at least until 24
Ma, with no evidence for any change in diversification rates even through the Cretaceous/Paleogene mass

extinction event. Given the ancient origin and stable environment provided by the TRF biome, comparatively old lineages have persisted and contributed to the steady accumulation of species diversity over time. In contrast to other plant studies, our results demonstrate that high species richness in TRF can be explained by a long and stable evolutionary process dating back to the mid-
Cretaceous.
Phylogeny and palm diversity across scales
Eiserhardt, WL 1
Couvreur, TLP 3
, Svenning, J-C
, Kissling, WD 1
1 , Baker, WJ
, Balslev, H 1
2 ,
1 Aarhus University, Denmark;
Kew, UK; 3
2 Royal Botanic Gardens,
The New York Botanical Garden, USA
Palms lend themselves to exploring approaches at the intersection of ecology and phylogenetics. They are taxonomically diverse, often key functional components of ecosystems, and form complex assemblages of different co-existing life-forms. Recent advances in palm phylogenetics, including a dated genus-level supertree covering all genera, allow new insights into the ecological and evolutionary processes that underlie this diversity. We used phylogenetic information and species distribution data to study the mechanisms behind diversity, composition, and structure of palm assemblages on different spatial scales. Among others, we ask the following questions: To what extent can niche conservatism explain large-scale distribution patterns?
Which assembly mechanisms are responsible for palm community composition on different spatial scales? What is the role of phylogenetic history for spatial patterns in functional assemblage structure? Our analyses are based on distribution data ranging from local to global scales:
(a) a dataset including >340,000 palm individuals in 430 transects in the Western Amazon, (b) a set of gridded distribution maps for all New World palm species
(n=550) and (c) global country-level presence/absence data of all (>2400) palm species. On a global scale, biogeographic regions show pronounced differences in the degree of palm phylogenetic clustering. The
Neotropics and a number of islands, notably Madagascar, but also e.g. New Caledonia, Hawaii, and Cuba, stand out with remarkably high clustering. On a continental scale in the New World, we inferred from phylogenetic turnover that palms diversified mainly within seven biogeographic regions. This pattern of in situ diversification is strongly driven by a combination of phylogenetic niche conservatism, environmental filtering and dispersal limitation. Niche conservatism with respect to temperature seasonality and extremes emerges as an important determinant of palm species and clade distributions and thus there is concern that palms might be considerably vulnerable to climate change. On a regional to local scale in the Western Amazon, preliminary results show phylogenetic overdispersion of palm communities (i.e., co-occurring species are less closely related than expected at random). This implies that community composition is influenced by interspecific competition (conserved alpha-niche) and/or adaptive speciation in response to spatially structured environmental factors (convergent beta-niche). Our findings indicate that the impact of environmental factors on the distribution of palm diversity can be understood much better in the light of phylogenetic history, challenging the traditional view that contemporary environment is the predominant driver of biodiversity patterns. Understanding how the distribution of species and thus biodiversity is influenced by the environment, and on which timescales these interactions take place, is urgent in times of anthropogenic climate change. It appears that phylogenetics can bridge a long-standing gap between ecology and evolution, and allow a holistic view on the causes of spatial biodiversity patterns.
Future of plastid DNA in palm phylogenetics
Asmussen-Lange, CB 1
1 University of Copenhagen, Denmark
The molecular era of palm phylogeny was initiated by
Uhl et al. in 1995 with their comprehensive family-wide study based on plastid RFLPs. This first plastid DNA phylogeny was followed by studies using DNA sequences from an increasing number of plastid DNA regions to resolve relationships among palm species at all levels from genus to family. At least 20 phylogenetic studies based on plastid DNA sequences have been completed, and 11 different plastid DNA regions have been explored. Heavy utilization of plastid DNA regions has given good results in higher-level studies and several lower level studies. How much more will this genome yield from sequences of targeted plastid DNA regions relative to the information and usefulness of whole plastid genome sequences? A DNA barcode for palms is one way of using specific plastid DNA regions because a
DNA barcode requires good quality single region sequences. Eight studies on palm phylogeny have used one or more of the proposed two-locus DNA barcode for plants, mat K and rbc L. The potential of DNA barcoding palms is explored by presenting new data on two groups of closely related palm species, the Geonomateae and the
Archontophoenicinae, and by reviewing published data on the barcode regions. The barcode data were evaluated using the criteria recommended by the Consortium for the Barcode of Life’s (CBOL) data standards and guidelines: universality, sequence quality and coverage, and discrimination ability. Other plastid DNA regions published for palms but not among the leading barcode regions were also evaluated. The performance of each
DNA barcode region varied extremely among palm groups, from being relatively discriminative to being almost identical among species in the group. In conclusion, some groups of palms can be DNA barcoded by sequencing a number of plastid DNA regions but not by the two-locus DNA barcode, mat K and rbc L alone.
Other groups of palms such as e.g., the Geonomateae and the Archontophoenicinae cannot be DNA barcoded by plastid DNA regions. Another important role of targeted plastid DNA sequencing is in assessment of whole plastid genome sequences. Here the existing or new sequences from specific plastid DNA regions may be used to explore data quality and amount of information of both targeted DNA region- and whole genome sequences. Also, as whole plastid genomes are being completed they may be integrated into existing plastid
DNA phylogenies to explore their usefulness in resolving relationships among palms. However, it is important to know the information quantity and quality of each targeted plastid DNA region. Data from a number of plastid DNA regions not yet explored in palm plastid
297

phylogeny will be presented and compared to existing data. Preliminary results indicate that several plastid
DNA regions have great potential for use in plastid DNA phylogeny.
Quaternary dynamics of Sundaland rainforest and its effects on current species distribution
Cannon, C 1 , Slik, F 1
1 Chinese Academy of Sciences, China
Sundaland forests have experienced a dynamic history through the Quaternary Period. Land area and connectivity among landmasses is strongly affected by sea level, with the entire shelf forming a large peninsula during glacial maxima. Paleoclimatic models and abundant off-shore stratigraphic evidence suggests that during glacial periods, while climate was slightly cooler and drier, precipitation and temperature levels were within the environmental envelope for evergreen rainforest over most of the shelf region. Current distribution of Sundaland rainforest therefore represents an extremely unusual and historically brief biogeographic setting while glacial distribution represents the more evolutionarily and ecologically significant biogeographic setting. The historical forests, due to differences in soilclimate-community settings, may have differed substantially in species composition and abundance from current forests. Additionally, different major forest types
(mangrove, lowland, and upland) have dramatically different histories, possibly explaining some of the correlation between ecological traits and community composition. Here, we examine the possible interaction among forest type, biogeographic history, and tree life history strategy to provide a rigorous framework for testing general assumptions about community assembly and the natural formation of refugia. We also use empirical data from a network of 111 tree plots from across Sundaland to test whether local historical dynamics can at least partially explain local species composition. Focusing on lowland forests, where species richness is greatest, we discuss possible mechanisms for the maintenance of diversity among suites of closely related, sympatric and interfertile species, including the positive impact of interspecific gene flow, against this historical background. Ultimately, the historical biogeography of Sundaland forests provide a compelling natural laboratory for understanding the effect of changes in climate and distribution on the evolution and ecology of diverse communities.
Pathways through Malesia van Welzen, PC 1 , Hovenkamp, PH 1 , Turner, H 1
1 Netherlands Centre for Biodiversity Naturalis, Branch
NHN, Leiden University, The Netherlands
The Malay Archipelago, also known as Malesia
(Malaysia up to the Philippines and Papua New Guinea),
298 has a very complex geological history. Most larger islands are complexes of amalgamated microplates and almost all microplates originated from the Australian plate or Australian part of Gondwana. The western half of Malesia (up to Borneo and a part of Sulawesi) broke off first and was already well in place before the second wave of microplates started to move away from Australia c. 50 Ma ago. For all modern taxa the western half of
Malesia is an extension of southeast Asia. Taxa that are widespread and cross Wallace’s line in the centre of
Malesia, could only have obtained their distribution after dispersal for which, theoretically, various routes are available. Phylogeographic data of Macaranga tanarius
(L.) Müll.Arg. (Euphorbiaceae), a widespread species ranging from S.E. Asia to Australia and the Pacific, combined with molecular phylogenies, dated or not, will have to show the general pathways through Malesia. The dated phylogenies will also be used to shed light on the discussion how early or which parts of the microplates emerged above water and could be used as stepping stones during dispersal. Geological evidence suggests a very late emergence of the central Malesian areas
(Sulawesi, Lesser Sunda Islands, Moluccas, parts of New
Guinea), mainly during the last 2 Ma, while phylogenies indicate that some parts were already above water at 25
Ma.
Glacial refugia of the Sunda Shelf and their impact on current species’ distributions
Raes, N 1
JWF 4
, Keßler, PJA
, van Welzen, PC 1
2 , Roos, MC 1 , Saw, LG 3 , Slik,
1 NCB Naturalis, The Netherlands;
Leiden, The Netherlands; 3
2 Hortus Botanicus
Forest Research Institute
Malaysia, Malaysia; 4
Garden, China
Xishuangbanna Tropical Botanical
For several temperate and tropical regions in the world, evidence suggests that species are still retracing their potential distribution since the ice sheets present during the last glacial maximum have retreated. For South-East
Asia information on the location of glacial refugia is at most scarce. Recent advances in this field of research suggest that due to much lower sea-levels and consequently larger land surface during the last glacial maximum, that it is today’s forests that are refugial, not those of the Last Glacial Maximum. However, how these cooler and dryer last glacial maximum climatic conditions have affected individual species’ distributions in not known. For this pilot study, we used digitized collection data from a selected number of plant families occurring on the Sunda Shelf (Borneo, Java, Malay
Peninsula and Sumatra) combined with current- and last glacial maximum climate data. Based on the assumption of ecological niche conservatism we hind-casted species distribution models developed under current climatic conditions to last glacial maximum climate conditions to identify the glacial refugia based on the selected species.
We further analyse how differences in the distribution ranges of species under present and past climatic conditions affect the contemporary botanical diversity pattern based on the selection of species.

Glacial soils on exposed Sunda Shelf shaped biogeographic patterns in Southeast Asia’s equatorial forests
Slik, JWF 1 , Aiba, S-I
Cannon, CH 0 , Eichhorn, KAO 7
Kartawinata, K 9
Marjokorpi, A
Nagamasu, H
Permana, A 21
17
12
2 , Bastian, M
, Fredriksson, G
, Laumonier, Y 10
, Meijaard, E 13
, Nilus, R 18
, Morley, R
, Nurtjahya, E
, Poulsen, AD 22 , Raes, N 23
0 ,
, Mansor, A
Brearley, FQ
19
14,15,16
, Payne, J
5
8
11
20
,
,
,
,
,
1 Keylab of Tropical Forest Ecology, Xishuangbanna
4
Tropical Botanical Garden, Chinese Academy of
Sciences, China; 2 Faculty of Science, Kagoshima
University, Kagoshima, Japan; 3 Philadelphia Zoo, USA;
Dept of Evolutionary Anthropology, Duke University,
USA; 5 School of Science and The Environment,
Manchester Metropolitan University, UK; 6 Biology,
Texas Tech University, Lubbock, USA;
Ecologie, Zeist, The Netherlands; 8
7 Eichhorn
Institute for
Biodiversity and Ecosystem Dynamics, University of
Amsterdam, The Netherlands; 9 Herbarium Bogoriense,
Research Center for Biology, Bogor, Indonesia; 10 Center for International Forestry Research, Bogor, Indonesia
Aim – The marked biogeographic difference between western (Malay Peninsula and Sumatra) and eastern
(Borneo) Sundaland has been explained by postulating a dry savanna dispersal barrier through the heart of the region. An additional explanation for this biogeographic pattern might be related to the coarse textured sandy soils of central Sundaland. Our aim was identify the filter
(climate and/or soils) that best explains current floristic patterns in the region. Location Borneo, Peninsular
Malaysia, Sumatra.
Methods – Using a data base containing 111 tree inventories (diameter at breast height (dbh) = 10 cm) we performed a floristic analysis at genus level. For each of the resulting clusters we determined the indicator genera and environmental correlates. Drought and coarse soil tolerance of the indicator genera of the floristic clusters that crossed the central Sundaland dispersal barrier were contrasted against those of non-crossing clusters.
Results – We found eleven terminal floristic clusters, ten occurring in Borneo, five in Sumatra and three in
Peninsular Malaysia. Indicator taxa of floristic clusters that occurred across Sundaland had significantly higher coarse soil tolerance than those from clusters that occurred either east or west of central Sundaland. For drought tolerance no such pattern was detected.
Main conclusions – We found that exposed sea bed soils probably acted as a dispersal barrier in central
Sundaland. We could not confirm the past presence of a continuous savanna corridor. During periods of intermediate sea levels, central Sundaland was likely covered by low diversity heath and herbaceous swamp forests that blocked the dispersal of tree taxa adapted to more fine textured and nutrient rich soils while enabling coarse sandy soil and possibly montane adapted species to pass. It also makes a coastal migration route for early humans more likely than one through central Sundaland as was previously proposed as part of the savannacorridor hypothesis.
Plant DNA barcodes: species identification and community phylogenies
Kress, WJ 1 , Erickson, DL 1
1 Botany, National Museum of Natural History,
Smithsonian Institution, Washington, USA
In plant communities species number, functional traits, and phylogenetic history all contribute to characterizing biological diversity. Until now the phylogenetic component of community diversity has been particularly difficult to quantify in species-rich tree assemblages. We have now applied DNA sequence data from plant DNA barcode markers to construct highly resolved community phylogenies for tree species in forest dynamics plots and have used these phylogenies to provide the evolutionary component for understanding community assembly. We used the plant DNA barcodes rbc L, mat K, and trn Hpsb A in a supermatrix approach along with phylogenetic constraint trees to generate community phylogenies in both temperate and tropical environments. The conserved barcode gene region rbc L along with the constraint trees provide the phylogenetic backbone at the family and ordinal level for the community analyses while the fasterevolving barcode gene regions mat K and trn Hpsb A permit the high resolution of the terminal twigs of the tree, i.e., the species. These results for forest dynamics plots in Panama, Puerto Rico, Maryland and Virginia illustrate how highly resolved phylogenies derived from
DNA barcode sequence data combined with constraint trees are particularly useful in comparative analyses of evolutionary diversity and enhance research on the interface between community ecology and evolution.
Divergence in anti-herbivore defense among sympatric
Psychotria
shrubs and implications for plant–insect interactions in phylogenetically-clustered
1 assemblages
Sedio, BE 1 , Hunter, MD 1 , Wright, SJ
University of Michigan, USA
1 , Dick, CW 1
Many tropical forests contain diverse assemblages of congeneric trees, which raises the question of how ecologically similar species can avoid competitive exclusion. Negative density-dependent fitness imposed by specialized natural enemies (i.e. the Janzen–Connell hypothesis) is a broadly accepted mechanism of species coexistence. Specialist herbivores, however, often specialize on a particular genus, not species, of plants.
On the other hand, plant defenses can be quite divergent among closely related sympatric species. Herbivoredriven spatial overdispersion of such defense traits might allow a Janzen–Connell-like mechanism to facilitate coexistence within local assemblages. We examine the specificity of interactions between herbivorous insects and their host-plants among 21 sympatric species of the diverse tropical shrub genus Psychotria (Rubiaceae) on
Barro Colorado Island (BCI), Panama. Data on plant– insect associations based on DNA barcode analysis of
299

insect gut contents are combined with profiles of host plant alkaloid secondary compounds to address three principal questions: (i) how specialized are the insect herbivores of a single, diverse plant genus? (ii) do
Psychotria alkaloid defenses show evidence of evolutionary divergence? and (iii) do Psychotria species assemblages reveal limited similarity in defense? Results indicate that the insect herbivores of Psychotria on BCI are not strict specialists. Phylogeny is a poor predictor of species alkaloid similarity in general, though particular principal components of alkaloid variation retain phylogenetic signal. While previous results indicate that species of Psychotria that co-occur within 28 m 2 plots are more closely related than by chance relative to the
BCI species pool (i.e. are phylogenetically clustered), cooccurring Psychotria are less similar chemically than by chance (i.e. are chemically overdispersed). Evolutionary divergence in defense may facilitate coexistence within such assemblages by reducing herbivore overlap between close relatives.
Developmental and functional tradeoffs affect interspecific diversification in the woody stems of the
Simaruba
clade of
Bursera
(Burseraceae)
Rosell, JA
Fornoni, J 3
1 , Olson, ME 1
, Domínguez, CA 3
, Aguirre-Hernández, R 2 ,
1 Instituto de Biología, Universidad Nacional Autónoma de México, Mexico; 2 Facultad de Medicina, Universidad
Nacional Autónoma de México; 3 Instituto de Ecología,
Universidad Nacional Autónoma de México, Mexico
Most biological structures peform multiple tasks, which translates into functional integration between organismal parts. When functions compete for the same developmental resources, patterns of negative covariation known as trade-offs emerge. Covariation patterns in general, and trade-offs in particular, have important implications for studies of adaptation, given that they can impose limits to natural selection and the phenotypes that can be observed. Trade-off studies have focused either on the population or macroevolutionary level and results are generalized to the other level, but studies are necessary to validate these generalizations. To study the effect of trade-offs in morphological diversification, linking population and interspecific levels, we studied woody stems. These structures are ideal simple systems performing just three main functions, mechanical support, storage, and water conduction. Using a morphologically diverse clade of tropical trees, the
Simaruba complex of Bursera (Burseraceae), we studied trade-offs between the main functions of the stem to understand integration in multitasking structures and its effect on interspecific morphological diversification.
Analyzing functional variables related to conductivity, mechanical support, and storage, sampling eight populations and nine species, we found strong functional integration in woody stems. In general, integration patterns were conserved between populations and also between species, suggesting an effect of functional integration in stem morphological diversification at the interspecific level. Trade-offs emerged mainly between variables related to mechanical support and storage.
Conductivity did not covary with other functions, but we did detect a trade-off between conductive safety and efficiency. Patterns of covariation were conserved
300 between species, but marked changes in mean performance values for each function were detected, and were associated with environmental conditions, suggesting that natural selection was responsible for these changes, in addition to being the causal agent maintaining the observed covariation patterns between functions. Functional integration affecting diversification in the simaruba clade could be representative of woody stems in general, given the pervasive nature of trade-offs in wood.
Searching for evolutionarily independent traits in tree structure
Olson, M 1 , Rosell, J 1 , Aguirre, R 2
1 Instituto de Biología, Universidad Nacional Autónoma de México; 2 Facultad de Medicina, Universidad
Nacional Autónoma de México, Mexico
The study of adaptation often focuses on 'parts' of organisms, with statement such as 'wings are an adaptation for flying' or 'vessels are adaptations for transporting water'. However, organisms develop and function as integrated wholes, making it unclear how and even if organisms should be 'atomized' into traits. A possible means of identifying traits is to study the patterns of covariation between parts, either within individuals throughout ontogeny, across the members of a species, or between species. The patterns of covariation, broadly known as phenotypic integration, may help identify organismal subunits that can change evolutionarily independently or semi-independently of other traits. Identifying semiautonomous organismal subunits and the causes of covariation and independence could help the quest for identifying biologically real
'traits'. For example, anatomists traditionally infer adaptive variation in xylem structural features as though anatomy interacted directly with the environment. The result is a canon of relationships, e.g. narrow vessels in drylands, wide in moist areas, etc., that has been widely accepted in comparative wood anatomy. However, vessel features can also be viewed as varying as a function of plant size, with natural selection acting to maintain hydraulic resistance constant, resulting in vessels that widen basipetally as their length increases. Plant size in turn is plausibly regarded being acted on by natural selection as a function of water availability and temperature, with plants being taller in warmer, moister areas. From this point of view, the traditional ecological wood anatomical canon is mostly a set of indirect relationships in which selection acts on plant size and anatomy is a response to size. We show that removing the effect of plant size eliminates the anatomy– environment relationship of many functionally relevant anatomical features, such as vessel diameter, an approach that also allows us to identify features that seem to vary independently of size. We suspect that many of the characters traditionally employed in ecological wood anatomy do not vary entirely independently of overall body size, and thus do not represent evolutionary characters in the sense of phenotypic units that are ontogenetically (quasi-) independent of other such units and that can respond directly to environmental selection pressure. As a result, the adaptive interpretations in this field may need to be revised.

The calcium oxalate crystals [COC] of some selected angiospermic taxa from India under the family
Araceae: used as an important taxonomic marker
Imam Saadi, SMA 1 , Mondal, AK 2
1 Vidyasagar University, India
In many plant species calcium oxalate crystals are commonly formed under ordinary conditions (Arnott and
Pautard 1970). These crystals are structural components in the leaves, stems, roots, storage organs and petiole of many higher plant families.We have studied calcium oxalate crystals (COCs) in the different plant parts
(leaves, stems, petiole, corm, and root) of some selected species belonging under the family Araceae. In the most studied plant types like marshy, semi aquatic, terrestrial and as well as the selected plant species are mostly herb, shrub, and climber in habit. Among the selected species two species are edible and economically important.
Edibility of petiole, leaves, stems, corm depends upon the frequency and intensity of the calcium oxalate crystals. We have observed two types of crystals which are mostly species specific. The low and high frequency of crystals is probably related with the habit, habitat and also the environmental conditions. The presence or absence of such important micro characters for understanding the evolutionary relationships of plant species (Franceschi and Horner 1980). The size and appearance of calcium oxalate crystals can differ within families, genus, and species and that these morphological characteristics are might be under genetically controlled.
The plant species always produces the same form of
COCs in the same tissue (Bouropoulos et al. 2001).
Analysis of genetic relationships on some species of
Cycadaceae in China by RAPD markers
Huang, Y-Y 1 , Nong, B-X 2
1 Zhongkai University of Agriculture and Engineering,
2 China; Key Lab of Guangxi Crop Genetic Improvement and Biotechnology, Academy of Agricultural Sciences,
China
Cycads are distributed in subtropical and tropical area plants, they are an ancient and rare plant group. Because some species of Cycadaceae still have some dispute in taxonomy, in order to address the issue, in this paper,
RAPD molecular markers were used to analyse the relationships among 21 species of Cycas . 21 random primers were identified with polymorphism among the entries. In total 333 bands were produced, of which 313 bands were polymorphic. Percentage of polymorphic bands was 93.99%.The data of 333 RAPD bands were used to generate Jaccard’s similarity coefficients and to construct a dendrogram by means of UPGMA in the
NTSYS-pc program. The results of RAPD cluster showed that the similarity coefficients of all species are ranged from 0.2903 to 0.7016, changing dramaticaly, showed the genetic background of Cycas is very complex. The similarity coefficient of Cycas multifida and C. segmentifida is 0.7016, which is the biggest one in that of all species, showed they have closest genetic relationship. The similarity coefficient of C. longlinensis and C. multifrondis was 0.2903, which is the smallest one in that of all species, showed they have farest genetic relationship. The similarity coefficient of different sex colony of C. diannanensis is 0.9098, which is the biggest one of all material, they cluster is first of all, indicated the closest genetic relationship of all material, and showed RAPD is suit for inter-species genetic relationship analysis of Cycadaceae. The results of
RAPD cluster provided important evidence to prove up the genetic relationship of all species of Cycas . Genetic relationship analysis by RAPD marker combining with morphology showed that C. multifida and C. segmentifida have close genetic relationship, C. multifida should be a subspecies of C. segmentifida . The species that have far genetic relationships of each other listed as follow: C. xilingensis, C. longlinensis, C. segmentifida,
C. acuminatissima, C.micholitzii, C. longipetiolula, C. multifrondis, C. longiconifera, C. guizhouensis, C. szechuanensis, C. brevipinnata, C. miquelii, C. balansae,
C. shiwandashanica, C. parvulus, C. fairylakea, C. hainanensis, C. taiwaniana, C. guangdongsensis, C. diannanensis , they are independent species from each other. We made a comparative analysis the ISSR markers on some cycad species in systematics with RAPD markers, the result showed that polymorphic bands amplified, percentages of polymorphic bands general are higher RAPD than ISSR, in the gene diversity among species (Ht) and within species (Hs) Ne, I, Ht and Gst,
RAPD and ISSR are very appropinquity, and some species are higher ISSR than RAPD, some species are
RAPD higher than ISSR, but their number difference are very small, their consistency exceed 90%.
Sapindales: an update on phylogenetics and age estimates
Muellner, AN 1 , Sapindales Phylogeny Group (SPG) 1
1 Biodiversity and Climate Research Centre (BiK-F) &
Goethe University, Germany
The Sapindales represent a diverse and economically important group of eudicots, with the APG system currently recognizing nine families: Anacardiaceae,
Biebersteiniaceae, Burseraceae, Kirkiaceae, Meliaceae,
Nitriaceae, Rutaceae, Sapindaceae (incl. Aceraceae and
Hippocastanaceae), and Simaroubaceae. Founded in July
2007 during the Botany & Plant Biology Joint Congress in Chicago, USA, the Sapindales Phylogeny Group
(SPG), aims at producing fully resolved, robust and dated phylogenetic trees to gain a deeper understanding of the evolutionary and ecological forces influencing diversification of this large order of mostly (sub)tropical flowering plants. The SPG, currently led by Alexandra
Muellner in close collaboration with Susan Pell and
Andrea Weeks, comprises experts of all nine families in
Sapindales, specialized in fields as diverse as floral morphology, taxonomy, systematics, DNA barcoding, phylogenetics, biogeography, and molecular clock dating. We here present the latest molecular systematic advances in the field, including phylogenetic analyses of more than 300 species, covering the most representative clades of all families in the order, and up to three plastid loci ( rbc L gene; atp B gene; trn L-F region, i.e. the trn L
301

intron plus intergenic spacer between the trn L intron and the trn F gene). In addition, we provide age estimates for all families, based on constraining multiple nodes in our phylogenetic trees by micro- and macrofossils. Our ages for sapindalean families are earlier than those obtained in more sparsely sampled analyses, although estimates for
Burseraceae agree surprisingly well. These and other molecular dating results highlight the need for dense taxon sampling as well as the importance of checking results against other types of evidence if time estimates are to be trusted. Sapindales, with their rich fossil record, provide an exceptional system for comparing different clock approaches because they permit such critical crossvalidation.
Trajectories of Anacardiaceae evolution: taxonomy, ecology, and structural diversity
Pell, SK 1 , Mitchell, JD 2
1 Brooklyn Botanic Garden, Brooklyn, USA;
York Botanical Garden, Bronx, USA
2 The New
The cashew family (Anacardiaceae) is an economically important, primarily pantropical eudicot family of approximately 800 species in 82 genera. The lineage reaches its greatest diversity in Malaysia and is most well known for its cultivated edible fruits and seeds (mango, pistachio, and cashew), dermatitis-causing taxa
( Melanochyla, Toxicodendron, Semecarpus , etc.) and lacquer plants ( Toxicodendron and Gluta spp.).
Collections from recent fieldwork in Papua New Guinea,
Vietnam, and New Caledonia have filled many gaps in our taxon sampling, which has resulted in a much more robust picture of Anacardiaceae evolution. Phylogenetic analyses of the greatly expanded dataset of nuclear (phyC and ETS) and chloroplast ( rps 16 and trn LF) DNA sequence data will be presented. These reconstructions and current knowledge of morphological and anatomical attributes of the Anacardiaceae support the recognition of modified versions of Takhtajan’s subfamilies
Anacardioideae and Spondioideae, as well as possibly a third as of yet unnamed subfamily segregated from
Spondioideae. In addition to elucidating the evolutionary relationships within Anacardiaceae, our research focuses on investigating the correlates of diversification in the family. Preliminary data have already shown that wind dispersal (a functional rather than morphological characteristic in this case because it involves different structures in different lineages) has evolved numerous times in the family and has a statistically significant correlation with ancestral colonization of dry habitats. In order to evaluate some of the other evolutionary trajectories in the family, we will consider structural adaptations and biogeography in the context of our elucidated phylogeny.
Tempo of evolution in
Acer
and
Aesculus
(Sapindaceae)
Harrington, M 1 , Biffin, E 2
1
2
Australian Tropical Herbarium, Cairns, Australia;
Discipline of Ecology and Evolutionary Biology, School of Earth and Environmental Sciences, The University of
Adelaide, Australia
302
Acer and Aesculu s (Sapindaceae) have a rich macrofossil record dating to the Paleocene. We present a variety of
Bayesian relaxed clock molecular estimates of divergence times for a combined plastid dataset ( rbc L, mat K) which either (1) incorporate the dates implied by the fossil record, (2) fossil constraints from outside
Sapindaceae, or (3) no fossil constraints. The results indicate contrasting evolutionary scenarios for Acer and
Aesculu s. Analyses with fossil constraints from outside
Sapindaceae imply a Pliocene–Miocene (6–28 Mya) origin rather than a Paleocene (ca 64 Mya) origin implied by the earliest attributed fossils, and an evolutionary rate for Acer and Aesculu s consistent with the majority of other genera of Sapindaceae sampled. Analyses including the Paleocene origin of Acer and Aesculu s as hard bound priors and four constraints from outside Sapindaceae result in a potentially biologically implausible rapid change in the mean evolutionary rate on the stem branch leading to the split between Acer and Aesculu s and their respective sister genera. In addition to questioning aspects of the hitherto accepted paleobotanical record, the conflicting scenarios presented suggest the need for a substantial re-evaluation of our understanding of the tempo and mode of evolution of these lineages.
Evolution of the New Caledonian flora: insights from the pantropical family Sapindaceae
Callmander, M 1,2
F 5 , Devey, D 5
, Munzinger, J
, Buerki, S 5
3 , Lowry II, PP 1 , Forest,
2
1 Missouri Botanical Garden, St Louis, USA;
Conservatoire et Jardin botaniques de la ville de
3 Genève, Chambésy, Switzerland; Institut de recherche pour le développement (IRD), UMR AMAP, Laboratoire de Botanique et de l'Ecologie Végétale Appliquées,
Herbarium NOU, Nouméa, New Caledonia; 4 Muséum
5
National d'Histoire Naturelle de Paris, France;
Molecular Systematics Section, Jodrell Laboratory,
Royal Botanic Gardens, Kew, UK
The economically important soapberry family
(Sapindaceae) comprises about 1900 primarily tropical species. On the southwest Pacific island of New
Caledonia, a globally recognized biodiversity hotspot with ca. 3,270 native species (ca. 75% endemic),
Sapindaceae have diversified significantly, comprising ca. 100 species in 13 genera (4 endemic), including many that remain to be described. Recent molecular phylogenetic studies indicate that at least four lineages colonized the island from the Eocene onwards, each diversifying to a varying degree. These analyses have shown that several genera with numerous representatives in New Caledonia (e.g. Cupaniopsis, Guioa ) are nonmonophyletic, which will necessitate significant changes in generic circumscriptions and numerous taxonomic realignments. In this study, the sampling of New
Caledonian representatives of the family is considerably expanded, now including almost all species currently assigned to Sapindaceae. We reconstruct the phylogenetic relationships within the family using a
Bayesian approach based on several plastid and nuclear
DNA sequence regions to elucidate the evolutionary history of the New Caledonian members of Sapindaceae.
State-of-the-art methods in molecular dating and reconstruction of biogeographical patterns are also used to infer the spatio-temporal evolution of the group on the

island. Our results will provide a foundation for broader analyses aimed at further testing the monophyly of geographically wide-ranging genera such as Cupaniopsis and Guioa , a key step toward developing a new treatment for Sapindaceae. The results will also contribute to ongoing investigations of patterns of diversification within the New Caledonia flora (in particular with respect to eco-geographic parameters such as bioclimate and substrate type) and will inform efforts to identify priority areas for conserving the island’s unique biota.
Multiple island radiations of Malagasy
Commiphora
(Burseraceae)
Gostel, M 1 , Weeks, A 1
1 George Mason University, USA
The myrrh genus, Commiphora (Burseraceae), has a rich cultural history, well known for fragrant oleoresins produced by its species. Less understood is the systematic biology of Commiphora , which contains approximately 190 species distributed throughout (sub-) tropical, seasonally dry forests or deserts in continental
Africa, Madagascar, India, Sri Lanka, the Middle East, and South America. Commiphora is best represented in continental Africa and Madagascar, which together are home to ca. 150 species. Previous molecular phylogenic studies have confirmed the monophyly of Commiphora and suggested no fewer than two dispersal events from
Africa to Madagascar, leading to the island’s ca. 50 endemic species. This study employs a much greater taxon sampling for Malagasy Commiphora derived from recent field collections, as well as an expanded DNA sequence dataset from two nuclear spacers (ITS & ETS) and two plastid spacers ( ndh Frpl 32 and psb Atrn H). We explore patterns of morphological trait evolution among species in each Malagasy subclade, establish the timing of their radiation and the probable geographic origin of their immigrant ancestors.
Age and historical biogeography of the pantropically distributed and species poor subfamily Spathelioideae
(Rutaceae)
Appelhans, MS 1, 2 , Janssens, SB 3 , Smets, E 1 , Keßler,
1
PJA 1
Netherlands Centre for Biodiversity Naturalis (section
NHN), Leiden University, The Netherlands; 2 Hortus
Botanicus Leiden, The Netherlands; 3 Laboratory of Plant
Systematics, Institute of Botany and Microbiology, KU
Leuven, Belgium
The subfamily Spathelioideae, which currently contains eight genera and about 30 species is a taxonomically difficult group. In their current circumscription the
Spathelioideae are extremely diverse in terms of morphology. As a result, earlier morphology based classifications placed only three of the genera in the
Rutaceae. Our molecular phylogenetic studies on five chloroplast markers ( rbc L, atp B, trn Ltrn F, rps 16, psb Atrn H) show that the Spathelioideae are the sister clade to the Rutaceae s.s. In addition, we found anatomical/morphological characters that support their placement in Rutaceae s.l. (glandular dots in leaves, specialised cells in the seed coat) and their fusion into one subfamily (oil cells in leaves, haplostemonous flowers, appendaged staminal filaments). The
Spathelioideae consist of two subclades of which one is strictly neotropical, whereas the other shows a palaeotropical distribution with one genus occurring in the Mediterranean and the Canary Islands. According to our molecular dating analysis, based on well identified and dated fossils of Clausena Burm.f. (Rutaceae) and
Ailanthus Desf. (Simaroubaceae), the splitting of the two lineages probably occurred in the Eocene and dispersal via the North Atlantic land bridge might have taken place. Further dispersal movements in the subfamily involve movements from mainland South America to the
Caribbean ( Spathelia ), from Africa to Madagascar
( Cedrelopsis ), and from Asia back to Africa
( Harrisonia ).
Unfurling fern biology in the genomics age
Barker, M 1
1 University of Arizona, USA
Among vascular plants, the economically important angiosperms have dominated genomic research efforts.
In addition to being directly used by humans for consumption, many flowering plant genomes are among the smallest for land plants and were amenable to early sequencing efforts. However, many of the most interesting questions in plant evolutionary genomics requires sequencing of the outgroups to flowering plants, including the gymnosperms, ferns, and lycophytes. These latter two clades of plants reproduce by spores and are often referred to as pteridophytes or seed-free vascular plants. These two monophyletic lineages possess a number of unique characters and occupy a phylogenetic position that make them vital for understanding the evolution and outstanding success of seed plants.
However, studying the genomes of seed-free vascular plants has proved to be difficult as their generally large genomes made the cost of sequencing unaffordable.
Recently, next-generation sequencing technologies, such as 454 and Solexa, are beginning to make genomic studies in pteridophytes a possibility. I will present the results of the first efforts of next-generation transcriptome and whole genome sequencing in ferns and lycophytes, and present a roadmap of forthcoming data sets and questions that may be tackled using such data in the coming decade.
Fern biogeography at multiple scales – the challenge of understanding fern specific processes
Korall, P 1
1 Uppsala University, Uppsala, Sweden
The distribution of organisms, at global, regional and local scales, and the processes behind it, are central questions in understanding biodiversity. Ferns disperse by spores and need water for fertilization. These two
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specific aspects of fern life history affect the chance of successful migration, and thus fern biogeography at multiple scales. However, these aspects have opposite effects. The small spores are easily transported by wind.
On the other hand, many ferns are outcrossing. In order to survive and reproduce in a new environment two spores have to land and develop fertile gametophytes close enough that a sperm produced in one gametophyte can swim to the egg in another. This is fundamentally different from how seed plants, the sister group to ferns, disperse and establish. To understand the patterns and processes shaping the biogeography of ferns, and hence their biodiversity, we need studies at different temporal, spatial, and phylogenetic scales across a broad sampling of lineages. Important is also an integrative, multidisciplinary approach involving historical biogeography, phylogeography, and ecology experiments, with well-supported phylogenies as a basis.
Ecophysiological, biochemical, and molecular mechanisms of desiccation tolerance in ferns
Watkins, JE 1 , Oliver, M 2 , Pruitt, N 3
1 Biology Dept, Colgate University, USA;
University of Missouri, USA; 3
2 USDA-ARS,
Colgate University, USA
The radiation of plants onto dry land required the evolution of adaptive character suites that allowed survival in a dry atmosphere. An extreme solution is to tolerate desiccation: to enter into and recover from an anhydrobiotic state. Early transitional land plants and many of their modern, yet basal, relatives like mosses and liverworts rely on desiccation tolerance (DT) to survive our deadly dry atmosphere. The goal of this work is to generate a better understanding of both the ecological and evolutionary significance of DT in ferns by examining molecular and physiological mechanisms of DT in this group. Specifically, we: (1) examine the extent and degree of tolerance between fern gametophytes and sporophytes using an array of physiological measurements, (2) understand how tolerance is related to the microenvironment of the species in question, and (3) elucidate the mechanisms of tolerance (between these two generations and across species of different habitats) by profiling genes that respond to dehydration and rehydration and the degree of their expression. We examined these three areas in the epiphytic gametophytes, the young epiphytic sporophtyes, and the soil rooted mature sporophtyes of the hemiepiphytic fern Lomariopsis vestita . As measured by chlorophyll fluorescence, we found that there are significant shifts in DT across this developmental gradient with epiphytic sporophtyes and gametophytes exhibiting greater degrees of DT than terrestrial rooted sporophtyes. These data suggest that there are significant life-stage mediated physiological shifts in tolerance that may help explain species distribution and the evolution of epiphytic ferns through hemiepiphytic intermediaries.
Phylogeny, physiology, and reticulate evolution: an integrated approach to North American
Dryopteris
(Dryopteridaceae)
Sessa, E 1 , Zimmer, E 2 , Givnish, T 1
2
1 University of Wisconsin-Madison, Madison, WI, USA;
Smithsonian Institution, Suitland, MD, USA
Despite the key position of ferns in land plant evolution, relatively little is known about the physiological traits that have driven their diversification, and few studies have combined molecular phylogenetic analysis with studies of physiological ecology to examine the causes of speciation and ecological diversification in a group for which the evolutionary relationships are well understood.
North American Dryopteris (Dryopteridaceae) is a model system for studying the roles of physiological adaptation and reticulate evolution in fern diversification, and our investigation integrates phylogenetic and physiological analyses to reconstruct evolutionary history and evaluate adaptive differences among taxa found in different light and water regimes. Dryopteris (the wood ferns) is a large, nearly cosmopolitan genus including ca. 225 species worldwide. The North American members of the genus constitute one of the most intriguing and beststudied groups of ferns worldwide, and have long been recognized as a classic case of reticulate evolution in plants. The North American complex includes 8 diploids and 5 fertile allopolyploids, which differ dramatically in morphology and habitat. Research over the last century culminated in a ‘reticulogram’ depicting putative relationships among these taxa, and invoked the existence of a missing diploid progenitor for two of the allotetraploids. Our preliminary phylogenetic results, based on analysis of ten maternally-inherited plastid markers and two biparentally-inherited nuclear markers, support all aspects of the reticulogram, including the existence of the missing ancestor. This research is nested within a broader study of phylogeny and historical biogeography of all New World Dryopteris , which has allowed us to identify non-North American taxa that appear closely related to the North American reticulate complex. Field and common-garden studies are being used to evaluate the potential value of key morphological and physiological differences among the polyploids and parental taxa, and integrating phylogeny will allow us to assess whether correlations remain significant when relationships are taken into account. We are especially interested in whether traits in polyploid hybrids are intermediate or transgressive relative to their putative parents, whether the tendency toward shorter, narrower fronds in species from sunnier habitats is adaptive based on photosynthetic sensitivity to low leaf water potentials and variation in light availability and transpiration rate, and whether leaf photosynthetic capacity is integrated with vein density, light availability, and frond orientation.
New insights on the phylogeny of Paleozoic ferns and implication on foliar organ evolution
Corvez, A 1 , Galtier, J 2 , Barriel, V 1 , Dubuisson, J-Y 1
2
1 UMR 7207 - MNHN CNRS UMPC, Paris, France;
UMR AMAP - CIRAD, Montpellier, France
The origin and evolution of the foliar organ, especially the megaphyll, is one of the major questions in the field of evolutionary developmental biology of land plants.
Because all living euphyllophytes (involving horsetails, ferns and seed plants) display megaphylls (except reversals such as in horsetails), a unique origin seems the
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a priori most parsimonious scenario. Conversely, paleobotanical data strongly support a multiple origin in monilophytes (horsetails and ferns) and lignophytes
(fossil progymnosperms and seed plants). Molecular evo devo analyses are mostly focused on living seed-plants.
The monilophytes and their fossils, often neglected in such studies, are nevertheless pertinent models for investigating the convergent evolutionary scenarii that would have provided the foliar organ in euphyllophytes.
In addition, a complete phylogenetic framework is strongly needed as a prerequisite to understand where and when the foliar organ and its potential precursors would have appeared. The evolutionary relationships among living monilophytes are currently well known, but few phylogenetic analyses involving fossil taxa were carried out. Therefore, the aims of this study were: 1) to reconstruct the phylogeny of monilophytes using morphology and anatomy data and selecting fossil and living representatives, more particularly Paleozoic fossil taxa of importance which have not been taken into account in any recent phylogenetic study, and 2) to infer on this phylogeny the evolution of anatomical and morphological traits that could contribute to the acquisition of the megaphyll. New insights into the relationships among basal monilophytes and new interpretations of such morphological and anatomical traits and of the definition of the foliar organ are thus discussed.
Paleobiological patterns in fern diversity
Nagalingum, N
1 University of California, Berkeley, USA; present address: National Herbarium of NSW, Sydney, Australia
The fossil record offers the opportunity to explore historical patterns that have shaped modern biodiversity.
The majority of living ferns diversified since the
Cretaceous, and molecular timetrees have helped elucidate the origin and timing of diversification of extant lineages. However, we are yet to understand the dynamics and timing of the demise of lineages that dominated the Mesozoic only to become minor components of extant floras. Using a data set of spores from the Cretaceous to Eocene, I examine trends in fossil fern diversity. I will present the diversity trajectories of ferns during and following the rise of angiosperms, showing for the first time measures of diversity that are statistically independent of the rise of angiosperms, thus providing more meaningful assessment of the decline in fern diversity. Diversification rates will be also be used to decipher the decline in ferns as either an increase in extinction rates (perhaps caused by the rise of angiosperms) or as a decrease in origination rates, which is consequentially independent of the radiation of angiosperms.
Stomatal control started as a passive response to water status
McAdam, SAM 1 , Brodribb, T 1
1 School of Plant Science, University of Tasmania,
Hobart, Australia
Little is known about the origins of stomatal control in vascular plants. Although angiosperms respond to a complex array of signals including light intensity, carbon dioxide concentration, humidity and leaf water status, phytohormones, toxic chemicals and pathogens, other land plant lineages (such as ferns and conifers) appear to lack a number of these responses. We are reconstructing the ancestral state and evolutionary development of stomatal control in vascular plants by combining analyses of gas exchange, hydraulic techniques and hormone quantification. Ferns and lycophytes were found to lack active stomatal closure in response to ABA and also hydropassive stomatal opening following leaf excision. The stomata of nine fern and lycophyte species examined did not close to any significant degree following a 90 minute feeding of 15000 ng ml -1 ABA into the transpiration stream, increasing foliar ABA levels in all fern and lycophyte species above 2000 ng g -1
FW. However, stomatal conductance (gs) in these basal lineages of vascular plants was highly sensitive to leaf dehydration and step changes in relative humidity.
Stomatal conductance rapidly declined following leaf excision in all fern and lycophyte species without evidence of hydropassive opening. We constructed a model that accurately predicted daytime gs in ferns and lycophytes based on leaf hydraulic parameters alone. In the three species examined ( Dicksonia antarctica ,
Pteridium esculentum and Lycopodium deuterodensum ) we observed gs consistently agreed with predicted gs in the steady state and transiently following changes in leaf water status. This 1:1 relationship between observed and predicted gs occurred despite significant variability in the half-time of stomatal kinetics as well as variability in the hydraulic parameters used to formulate the model. This suggests that the stomata of ferns and lycophytes act as passive hydraulic valves. As ferns and lycophytes represent distinct basal vascular plant lineages these results indicate that passive hydraulic stomatal control represents the ancestral state of vascular plants. The control of gs by carbon dioxide concentration and the enhancement of this signal by increased ABA level was assessed in conifers. Increased ABA level following drought did not enhance the stomatal response to changes in carbon dioxide concentrations in conifers. This initial evidence suggests that conifers posses an intermediate state of stomatal control between that of the passively controlled stomata of ferns and lycophytes and the predominantly active biochemically controlled stomatal processes in angiosperms. These results provide important insight into the evolution of stomatal control, with most of the active processes governing stomatal behaviour in angiosperms derived long after the divergence of ferns 360 million years ago. The ancestral state of stomatal control in vascular plants appears to be one of an active response to red light and a passive response to leaf water balance. This suggests that until the rise of the conifers, global scale biotic fluxes of CO and H
2 relative humidity and light intensity.
2
O were dependent only on soil water content,
At the fern frontier: new insights into the causes and consequences of variation in fern leaf form and function across species and environments in Southern
California, Hawaii and Costa Rica
Creese, C 1 , Sack, L 1
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1 University of California, Los Angeles, USA
Ferns are an excellent system to test hypotheses of adaptive trait relationships as they exhibit the greatest diversity in form and habit, are the most speciose vascular plant group after angiosperms with 12000 species, and range widely across environments, from semi-deserts to rainforests. Ferns provide a means to test the universality of trait relationships established for angiosperms and gymnosperms and are revealing new trait relationships that help to explain the evolution of plant diversity. Since leaves are the metabolic engines of plants, identifying what drives variation in their form and physiology is especially critical to elucidate patterns of evolutionary diversification and ecosystem function.
Focusing on leaves from 15 morphologically diverse species growing across environmental gradients in
Southern California, we tested suites of hypotheses concerning the size-scaling of plant parts, leaf economics, functional consequences of leaf shape and venation architecture, trait associations with growth form and environment, and identified novel structure-function relationships that depart from trends reported for angiosperms. Within and across fern floras, we found geometric scaling of plant parts to maintain functional proportions and a strong association of plant size with growth form. For Southern California ferns, leaf size, shape and composition as well as hydraulic conductance and photosynthesis varied significantly with moisture availability and irradiance. Key structure-function relationships shared with angiosperms and gymnosperms included leaf economic trait associations, such as the trade-off between leaf mass per area and photosynthetic rate. Novel relationships included the decoupling of vein density, leaf hydraulic conductance and photosynthesis in dry, high irradiance environments. Results draw attention to adaptive limitations and innovations in ferns and identify where research is needed to expand our understanding of fern biology and their contribution to ecosystem function.
Phylogenetic relationships and diversification patterns of the Lindsaeoid ferns
Lehtonen, S 1 , Tuomisto, H
Christenhusz, MJM 3
1 , Wahlberg, N 1 , Rouhan, G 2 ,
1 University of Turku, Finland; 2 d'Histoire Naturelle, Paris, France; and Herbarium, Helsinki, Finland
Muséum National
3 Botanic Garden
Polypod ferns represent more than 80% of the living fern species, and form a well supported clade in all recent phylogenetic analyses. Two main clades exists within the
Polypodiales – one clade containing the lindsaeoid ferns and a few other small genera, the other and much larger clade being composed of the pteridoid, dennstaedtioid and eupolypod ferns. Molecular data together with palaeobotanical evidence suggests that the division between these two clades occurred in the Late Jurassic, and major diversification among the main polypod groups began during the Late Cretaceous. However, the sampling within the lindsaeoid clade has remained poor in most phylogenetic studies and it remains unclear whether the lindsaeoids and associated genera really form a clade, or if they form a grade from which the pteridoid-dennstaedtioid-eupolypod lineage emerged. As
306 well, the age of the crown-group lindsaeoids and time of their further divergence has remained obscure. Our recent studies have clarified the phylogenetic history of the lindsaeoid ferns. However, despite using large amounts of molecular data and various analytical approaches, we have not been able to confirm the exact phylogenetic position of Cystodium and Saccoloma , two enigmatic genera nowadays usually associated with the lindsaeoids.
Based on relaxed molecular clock analyses it appears that, even though the original split between the lindsaeoid and non-lindsaeoid Polypods occurred already at the end of the Jurassic, the crown-group lindsaeoids did not diversify until Cenozoic. Thus, the independent diversification of the lindsaeoid and non-lindsaeoid
Polypods occurred more or less simultaneously. The current distribution pattern within the lindsaeoids seems to result from rare long-distance dispersal events followed by speciation events. Interestingly, the
Pantropical and most species-rich lindsaeoid genus
Lindsaea is almost completely absent from the continental Africa, and appears to have a surprisingly short history in the Amazon basin. Especially the rapid and apparently adaptive radiation in Amazonia requires further attention. The authors declare no conflict of interest.
1
Systematics of Australasian Aspleniaceae
Ohlsen, D 1
The University of Melbourne, Australia
Asplenium is Australia's largest fern genus, with over 30 species occurring on mainland Australia. A molecular phylogeny for the genus has been established largely with species occurring outside of Australia. This study is the first to sequence all species of the genus in Australia, for the chloroplast regions rbc L, trn L-F and rps 4-trnS.
Maximum parsimony trees were generated, incorporating this new data, and demonstrated that five of the eight main clades of Asplenium occur in Australia, with two of these clades being particularly well represented. These trees also prove to be valuable for assessing the number of taxa that should be acknowledged within Australia.
Phylogeny and rarity in Australian 'tassel-ferns'
Field, A 1 , Holtum, J 1 , Waycott, M 1
1 School of Marine and Tropical Biology, James Cook
University, Port Douglas, Australia
The Lycopods are the most ancient surviving vascular plant phylum, with a cosmopolitan fossil record extending back 400 million years. The most plesiomorphic genus of lycopod is the broadly defined and heterogeneous Huperzia , which contains the temperate terrestrial ‘fir-mosses’ and the tropical rainforest epiphyte ‘tassel-ferns’. The phylogenetic diversity among the 12 Australian Huperzia is unknown but close relationships with Melanesian taxa are suspected. All of the Australian tassel-ferns are rare and threatened and at least one, the blue-tassel-fern, is likely to become extinct in the wild in Australia in the near future. An accurate phylogenetic systematic arrangement is needed to decide which taxa are truly rare by determining whether cryptic taxa are present or whether

undiagnosed relationships with common populations outside of Australia are present. This study of the most comprehensive genetic and living collection of tasselferns available tests the ‘in-use’ taxonomic hypothesis as well as recent phylogenetic hypotheses of the
Lycopodiaceae by using Bayesian inference phylogenetic analyses of the chloroplast loci trnL C-F, trnH-psbA and aCCD in addition to 85 non-DNA characters obtained from plants grown under controlled conditions. Two subfamilies within Lycopodiaceae are strongly supported but the genus Huperzia is polyphyletic and paraphyletic.
A monophyletic arrangement is obtained if the fir-mosses
( Huperzia s.s.) are segregated from the tassel-ferns
( Phlegmariurus broadly defined) and the pygmy clubmoss ( Phylloglossum ). Based on phylogenetic analyses, four additional Australian taxa are recognised and two hitherto considered as occurring in Australia are eliminated. A revised assessment of the rarity of the taxa in Australia increases the number of vulnerable taxa by three and also increases the number of endemic taxa.
Most of the remaining Australian tassel-fern species range into Melanesia and share their closest relatives with populations overseas. The endemic taxa are limited to high altitude rainforests of the Atherton Wet Tropics bioregion. The critically endangered blue-tassel-fern is basally positioned within the tassel-fern clade and is not closely related to any other taxon. The blue-tassel-fern exhibits a unique morphology and specialised hostdependency that is considered to be one of several potential causes of rarity for this species.
Evolutionary history and biogeography of
Tecophilaeaceae
Forest, F 1 , Manning, JC 2
1 Royal Botanic Gardens, Kew, UK; 2 Compton
Herbarium, South African National Biodiversity Institute ,
South Africa
Tecophilaeaceae is a small family comprising 9 genera and 24 species of geophytic perennials, characterised by a cormous rootstock, poricidial anthers and semi-inferior ovary. Most of the genera and species are found in sub-
Saharan Africa ( Cyanastrum , 3 spp.; Cyanella , 7 spp.;
Eremiolirion , 1 sp.; Kabuyea , 1 sp.; Walleria , 3 spp.), while Conanthera (5 spp.), Tecophilaea (2 spp.) and
Zephyra (1 sp.) are restricted to Chile and Odontostomum
(1 sp.) is endemic to California. This disjunct distribution in North and South America and Africa poses an interesting problem: what are the potential biotic or abiotic factors that may explain these distributional patterns and which processes would have influenced them (e.g. long-distance dispersal). In order to address this question, and to review the taxonomy and systematics of the group, phylogenetic relationships within the family were reconstructed using maximum likelihood and Bayesian methods and six plastid DNA sequence regions. The phylogenetic relationships uncovered show strong biogeographical clustering, with the Chilean species grouping together and sister to the remainder of the family, in which the North American
Odontostomum is sister to the sub-Saharan species.
Within the African clade, the tropical genera Cyanastrum and Kabuyea are sister to the southern African genera
Eremiolirion , Cyanella , and Walleria (the latter also found in tropical regions). Relationships are generally well supported and all genera are found to be monophyletic, except for Tecophilaea , in which Zephyra is nested. The stem lineage of the group is thought to have evolved about 105 million years ago, which would suggest a Gondwana origin for the group. The evolutionary history of this group and the possibility of a
Gondwanan origin is explored further using divergence time estimates provided by a Bayesian relaxed clock and biogeographical patterns assessed obtained using stateof-the-art ancestral area reconstruction methods (e.g.
Bayes–DIVA, Lagrange).
Molecular systematics of the aloes (
Aloe
L.,
Xanthorrhoeaceae) – progress towards a predictive phylogeny
Grace, O 1, 2 , Bjora, C 3 , Forest, F 2 , Rønsted, N 1
1 University of Copenhagen, Denmark;
Gardens, Kew, UK;
2 Royal Botanic
3 Natural History Museum, Oslo,
Norway
The systematics of the utility genus Aloe L.
(Asphodelooideae – Xanthorrhoeaceae) is an emergent area of study in an otherwise well-characterised group of considerable diversity (ca. 600 species) as well as taxonomic complexity. Molecular characters hold promise for resolving artificial elements in the classification of the genus, yet phylogenies generated to date from DNA sequence data have conflicted with the morphogroups of existing classifications of Aloe , and have only provided limited resolution. In the present study, we tested a series of DNA regions among alooid taxa, comparing plastid regions such as mat K, rbc L, the trn LF intron and spacer, rpo C1 and psb Atrn H, and the nuclear regions ITS and phyC. The most informative of these are being used to assemble a comprehensive phylogeny of Aloe , representing the full morphological and geographical diversity of the genus. We are significantly expanding upon the number of taxa and regions sequenced to cover about 25% of the genus.
Recent progress towards a well resolved phylogeny of
Aloe will be presented. Insights into evolutionary relationships among aloes will be useful in recovering a natural classification of the genus, since the most recent classification (published in two volumes by G.W.
Reynolds in 1950 and 1966) remained incomplete and lacked predictive power. A robust phylogenetic hypothesis has several applications beyond the infrageneric classification of Aloe . We intend to explore the phylogenetic approach to pinpointing priority taxa for conservation as well as for natural products research. A genus such as Aloe characterised by high species diversity and endemism, as well as cultural and economic value, presents substantial challenges to achieving conservation and sustainable use. The relevance of molecular systematics in achieving these in Aloe will be highlighted here.
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A revision of
Aloe
L. section
Purpurascentes
Salm-
Dyck, using morphological, biogeographical and phylogenetic data
Klopper, RR 1 , Grace, OM 2 , van Wyk, AE 3 , Smith, GF 3
1 Biosystematics Research & Biodiversity Collections
Division, South African National Biodiversity Institute,
South Africa; 2 Jodrell Laboratory, Royal Botanic
Gardens, Kew, UK; Marie Curie Fellow, Dept of
Medicinal Chemistry, University of Copenhagen,
Denmark; 3 HGWJ Schweickerdt Herbarium, University of Pretoria, South Africa
The genus Aloe L. comprises ± 600 species from Africa, the Arabian Peninsula, Madagascar and some Western
Indian Ocean Islands. Southern Africa is one of the centres for aloe diversity and harbours ± 150 taxa. Aloes are valued for a multitude of traditional biocultural uses, and are immensely popular among succulent plant collectors and horticulturalists. Many aloes are threatened by habitat destruction and illegal collecting.
Aloe Section Purpurascentes Salm-Dyck currently comprises five species from South Africa and southern
Namibia: Aloe framesii L.Bolus is confined to sandy coastal flats on the northwest coast of South Africa and is regarded as Near Threatened; A .
gariepensis Pillans is endemic to the lower Orange River valley in northwestern South Africa and southern Namibia; A. khamiesensis Pillans is restricted to a relatively small area in the Succulent Karoo of South Africa; A. microstigma Salm-Dyck has a disjunct distribution, occurring fairly widespread in the western and southern regions of South Africa, as well as in southern Namibia;
A. succotrina Weston is confined to mountain fynbos in the Western Cape, South Africa. Several questions regarding the demarcation of species in this section exist, especially concerning the disjunct populations of A. microstigma and the status of A. khamiesensis and A. framesii . The Namibian population of A. microstigma was described under the name A. juttae Dinter in 1923, but was subsumed under A. microstigma in 1974. Aloe khamiesensis was included as a synonym of A. microstigma subsp. microstigma in 2000, and A. framesii reduced to subspecific rank [ A. microstigma subsp. framesii (L.Bolus) Glen & Hardy]. These views, however, remain controversial. There are also distinct populations of aloes belonging to this section that warrant possible description as new taxa. A taxonomic revision of the section is being conducted under the auspices of the Aloes of the World Project, which forms part of the African Plants Initiative, supported by the
Andrew Mellon Foundation. The main objective of the
Aloes of the World Project is to provide an electronic facility with all relevant information and available images on this genus on a world scale. The revision of
Aloe section Purpurascentes aims to resolve the species and infraspecific circumscriptions of the group. This is done through field studies, evidence from morphology, ecology and biogeography, as well as DNA-based phylogenetic analyses. The data gathered are combined to produce a revised taxonomic treatment of the taxa involved, including full descriptions following the comprehensive Aloes of the World description template.
The results of this and other research done by components of the Aloes of the World Project will enhance our understanding and general knowledge of this
308 important genus. It will also enable environmental decision-makers to improve the conservation of members of the genus.
Phylogeny and evolution of Orchidaceae: a phylogenomic perspective
Ames, M 1
M 2
, Williams, N
, Neubig, K
2 , Leebens-Mack, J
2 , Clements, M 4 , Givnish, T 1
3 , Whitten,
1 Dept of Botany, University of Wisconsin, Madison,
USA; 2 Florida Museum of Natural History, University of
Florida, USA; 3
Georgia, Athens;
Dept of Plant Biology, University of
4 Centre for Plant Biodiversity Research
CSIRO Plant Industry, Australia
Orchids, one of the largest families of flowering plants, include about 900 genera and 25,000 species, representing nearly 10% of all angiosperm species. Most of them inhabit tropical forests, with 80% of all orchid species being epiphytes. Orchids who show remarkable specializations in reproductive biology like the labellum and the pollinia as well as their reproductive success based on deception have fascinated evolutionary biologists since Darwin and they continue to be the focus of evolutionary studies. The understanding of orchid relationships at a variety of levels within the family has greatly improved in the last 15 years through the use of molecular data. Studies based on one to five plastid genes have identified five subfamilies related in ladder fashion, including Apostasioideae, Vanilloideae, Cypripedioideae,
Orchidioideae, and Epidendroideae. As part of the
Monocot AToL project, we and our colleagues are sequencing whole-plastid genomes to provide a broadscale phylogeny for the monocots as a whole, and to investigate detailed relationships within some of the largest families, including the orchids. Our analysis places order Asparagales sister to the commelinids, with
Orchidaceae sister to all other families of Asparagales. In order to provide a solid backbone phylogeny for
Orchidaceae, we are sequencing plastomes for species representing all tribes and several subtribes in each of the subfamilies. Our presentation will focus on the implications of the plastome phylogeny for relationships and evolutionary patterns within the subfamily
Epidendroideae, which includes 80% of orchid species, and almost all epiphytic orchids. Relationships within the subfamily have, however, remained rather enigmatic based on analyses of one to a few plastid genes, with apparently very rapid diversification in several groups and very short, poorly supported branches. Our data cast new light on relationships among the epidendroid tribes and the apparent tempo at which they diverged from each other.
Generic relationships within Asparagaceae:
Lomandroideae inferred from non-coding chloroplast
DNA and morphology
Sirisena, UM 1, 2 , Donnon, MJ 1 , Macfarlane, TD 2 ,
1
Conran, JG 1
Australian Centre for Evolutionary Biology and
Biodiversity, School of Earth and Environmental
2
Sciences, The University of Adelaide, Australia;
Western Australian Herbarium, Manjimup Research
Centre, Australia

A phylogenetic analysis using trn L intron and trn L–F intergenic spacer sequence data and morphological data was carried out in order to reassess the generic relationships of Laxmanniaceae and to investigate the validity of the two putative informal generic groups. In all analyses, three clades were recovered: the blackseeded laxmannioids ( Laxmannia R.Br. and Sowerbaea
Sm.) and arthropodioids (as a pair), sister to the nonphtytomelanic lomandroids. The laxmannioids possess bracteate, pedunculate flower heads and ligulate leaf bases while the lomandroids were defined clearly by characters including: the absence of tubers or phytomelanic seeds and the presence of long-lived flowers with few ovules. In contrast, the arthropodioids only apparent synapomorphy was multi-ovulate locules, but this character also occurs in at least some species of both Laxmannia and Sowerbaea . Tentatively placed taxa such as Murchisonia Brittan and Trichopetalum Lindl. were nested inside the family with strong bootstrap support, but Murchisonia is polyphyletic and nested inside Thysanotus R.Br. Similarly, Xerolirion
A.S.George is apparently a highly-derived Lomandra
Labill. and there are problems with relationships between the various species of Chamaexeros Benth. and
Romnalda P.Stevens.
Differential expression of paralogous DEFICIENS- and GLOBOSA-like MADS-box genes in the flowers of Orchidaceae: refining the orchid code
Mondragón-Palomino, M 1
1 University of Regensburg, Germany
Goethe considered orchids as 'monstrous lilies', while
Darwin and others hypothesized about the mechanisms behind their particular flower morphology. However, relatively little is known about the genetics of orchid morphological diversity. Our project, approaches this question by determining how changes in the number, sequence and expression of developmental regulatory genes resulted in the formation of orchid-specific flower structures. In angiosperms class B floral homeotic genes encode MADS-domain transcription factors which are key in the specification of petal and stamen identity and have two ancient clades: DEF-like and GLO-like genes.
Many species have one gene of each clade, but orchids have typically four DEF-like genes, representing ancient gene clades 1, 2, 3 and 4. Based on the molecular phylogeny of DEF-like genes and the patterns of expression documented in the literature, we developed the 'orchid code'. This model considers the identity of the perianth organs is specified by the combinatorial interaction of four DEF-like MADS-box genes. We tested the hypotheses derived from the 'orchid code' by characterizing the patterns of evolution and expression of the genes we isolated from several species. Here, I discuss these computational and experimental analyses and consider the contribution of gene families to the emergence of morphological novelties during evolution.
Molecular phylogenetics of Asparagales based on nuclear, plastid and mitochondrial DNAs 7 coding gene sequences
Kim, J-H 1 , Kim, D-K 1 , Choi, J-E 1 , Yu, H 1 , Chase, MW 2
1 Kyungwon University, Seongnam, Korea;
Laboratory, Royal Botanic Gardens, Kew, UK
2 Jodrell
A phylogenetic analysis of a combined data set for 400 taxa of Asparagales based on seven DNA coding genes,
18S rDNA(1796bp), plastid rbc L(1368bp), mat K(1668bp), ndh F(2040bp), atp B(1420bp), ycf 5(375bp) and mitochondrial mat R (1860bp), and representing a total of ca. 11.0kb is presented. Parsimony and Baysean inference analyses were conducted to elucidate the relationships of Asparagales and related groups, and bootstrap analysis was performed to assess the support of clades. The combination of seven coding genes data set has resulted in the most highly resolved and strongly supported topology yet obtained for
Asparagales. Among the seven coding genes, plastid matK and ndhF gave more informative contribution. The relationships of these results were nearly congruent with that by Chase et al. (1995), APG, APG II and APG III.
And parsimony and Bayesian analyses yielded identical relationships except slightly among core asparagoids.
The core asparagoids sensu Chase et al. form a strongly supported monophyletic group, whereas the non-core asparagoids form a paraphyletic clade. In the core asparagoids, five clades were grouped: (1) Alliaceae s.l.(sensu APG II, Amarylidaceae–Agapathaceae–
Alliaceae), (2) Laxmanniaceae–Asparagaceae–Ruscaceae s.l. (3) Themidaceae, (4) Hyacinthaceae, (5)
Anemarrhenaceae–Behniaceae–Herreriaceae–Agavaceae s.l. Clades (2), (4) and (5) form a strongly supported clade, and clades (1) and (3) show a moderately supported clade. The position of Aphyllanthes is labile according to the genes, but it is joined with Themidaceae in the combined tree. The highly supported monophyletic clade of Xanthorrhoeaceae s.l. (sensu APG II, including
Asphodelaceae and Hemerocallidaceae) in the lower asparagoids is the sister group of all the higher asparagoids. And the phylogenetic position of lower asparagoids is slightly different from the result of Fay et al. (2000). The successive sister groups to the higher asparagoids were: Xanthorrhoeaceae s.l.;
Xeronemataceae; Iridaceae; Ixioliriaceae–Tecophilaceae;
Doryanthaceae; Asteliaceae–Lanariaceae–Boryaceae–
Blandfordiaceae; and Orchidaceae. Parsimony and
Bayesian analyses yielded identical relationships except for some slight variation among the core asparagoid families, which nevertheless form a strongly supported group. The core asparagoids fell into two clades. The first consisted of four families including Agavaceae s.l. sensu APG, Hyacinthaceae, and Themidaceae as well as
Aphyllanthaceae. The second consisted of Ruscaceae s.l.,
Asparagaceae, Laxmanniaceae, Alliaceae, Agapathaceae, and Amaryllidaceae. Also, this study confirmed monophyly of Ruscaceae s.l. with strong support and it also supports the inclusion of Eriospermaceae.
Relationships within Asparagales: a phylogenomic perspective
Steele, PR 1 , Hertweck, K
, McKain, M 2 , Pires, JC 1
1 , Docktor, V 1 , Leebens-Mack,
1
J 2
University of Missouri, Columbia, USA; 2 University of
Georgia, Athens, USA
309

Except in a few notable phylogenetic studies, current methods of estimating evolutionary histories of plant groups and identifying species with DNA sequences involve only a few genes or noncoding regions from the chloroplast genome and, sometimes, the nuclear genome.
These methods are not always repeatable across distantly related angiosperms. The present study utilized state-ofthe-art, high-throughput sequencing technology and the latest bioinformatics tools to infer the phylogeny of core
Asparagales. This process did not require labor-intensive chloroplast isolations or PCR cycle or reagent variations.
It included a kit-based extraction of total genomic DNA and a kit-based sample preparation for massively parallel sequencing. Results included nuclear and mitochondrial gene sequences, and whole chloroplast genome sequences, representing dozens to hundreds of alignable genes and noncoding regions. These data from multiple sources provided the opportunity to estimate a wellsupported phylogeny of core Asparagales, discover the phylogenetic position of the monotypic genus
Aphyllanthes , and compare and contrast the evolution of genes from independent datasets. The majority of angiosperms contain chloroplast genomes that have the same structure, genes, and gene order – commonly called the 'typical' chloroplast genome. Our results revealed the loss of a few genes from the chloroplast genome in some
Asparagales plant groups as compared to the typical land plant chloroplast genome, and they revealed the most variable regions. These sequences can be used to investigate the evolution of genes, morphological traits, and biogeographic history. This study focused on plants, but we tested this novel approach to DNA sequencing on other groups of organisms such as insects and mammals, for which we sequenced whole mitochondrial genomes using the same methods. A brief review of these results will also be presented.
Origin of bimodal karyotype of Agavaceae coincides with a paleopolyploid event
McKain, MR
CW 3
1 , Pires, JC
, Leebens-Mack, JH 1
2 , Wickett, NJ 3 , DePamphilis,
1
2
Dept of Plant Biology, University of Georgia, USA;
Div. of Biological Sciences, University of Missouri,
USA; 3 Dept of Biology, Penn State University, USA
Polyploidy is a common phenomenon in angiosperm history. Evidence suggests that most if not all angiosperm lineages have experienced multiple polyploidy events in their evolutionary history. The timing of these polyploidy events relative to bursts of diversification and speciation in the angiosperm tree of life has lead many to hypothesize a causal link. For example, members of Agavaceae sensu stricto possess a bimodal karyotype that has been hypothesized to be the consequence of a paleopolyploid event predating the diversification of major lineages in the family. We tested the hypothesis that the origin bimodal karyotype is coincident with an ancient polyploidy event by analyzing sequenced transcriptomes from six Agavaceae sensu lato species ( Chlorophytum rhizopendulum, Hosta venusta,
Agave attenuata, Yucca filamentosa, Hesperaloe parviflora and Chlorogalum pomeridianum ). Frequency plots for synonymous substitutions (Ks) between putative ortholog and paralog pairs were constructed and inspected for evidence of ancient polypoidy. The Ks
310 plots suggest that all species with the Agavaceae s.s. share a paleopolyploid event that occurred just before the earliest branching events in the family. A separate polyploidy event is implicated for the lineage leading to
Chlorophytum (Anthericaceae). These inferences were tested in phylogenetic analyses of gene families using genes mined from transcriptome data from Agavaceae,
Chlorophytum, and the more distantly related taxa
Asparagus and Aloe . Divergence times were estimated for the two paleopolyploid events and the implications of these dates have been explored with regard to karyotype and diversification.
The Astelioid clade of Asparagales: molecular phylogenetics, character plasticity and long-distance
1 dispersal
Kocyan, A 1 , Birch, JL 2
University of Potsdam, Institute of Biochemistry and
Biology, Biodiversity Research/Systematic Botany,
Germany; 2 University of Hawaii at Manoa, Dept of
Botany, Honolulu, USA
The almost exclusively Southern Hemisphere astelioid clade (Asparagales) contains four families, the
Asteliaceae, Blandfordiaceae, Hypoxidaceae and
Lanariaceae. An additional endemic Australian family,
Boryaceae, is sometimes included in the astelioid clade or placed as the next family in the basal Asparagales grade. Astelioid relationships have been proposed based on large scale Asparagales studies, yet without broad sampling across the diversity of these families understanding of astelioid evolution and biogeography remains incomplete. Within the Asparagales, the
Orchidaceae are sister to a clade in which the astelioids are sister to all other Asparagales. Therefore, knowledge of the extensive morphological diversity of the astelioid clade is essential for understanding Asparagales evolution. The first molecular phylogeny of all currently accepted genera of astelioids, including Boryaceae, is presented. Phylogenetic reconstruction is based on the plastid DNA regions ( rbc L, trn Ltrn L-F, trn S-G, psb Atrn H, rps 16trn K, and pet Lpsb E). Preliminary results indicate that Boryaceae and Blandfordiaceae are successive sister to the clade containing
Asteliaceae/Hypoxidaceae /Lanariaceae. These relationships suggest extensive plasticity of vegetative and floral characters. Septal nectaries, which are external in Blandfordiaceae and Boryaceae and internal in
Lanariaceae and Asteliaceae, appear to have been secondarily lost in the Hypoxidaceae. Multiple transitions in ovary position appear to have occurred: the ovary is superior in Boryaceae and Blandfordiaceae, inferior in Hypoxidaceae, and superior to semi-inferior in
Asteliaceae and Lanariaceae. Multiple transitions between fleshy and dry fruit and unilocular and trilocular ovaries have occurred in both the Asteliaceae and the
Hypoxidaceae. Astelioid distributions on all four
Gondwanan landmasses are consistent with a
Gondwanan origin of these families and estimation of divergence times for the major astelioid clades is underway to investigate congruence with Gondwanan biogeographic hypotheses. Long-distance dispersal has clearly influenced diversification of Asteliaceae and
Hypoxidaceae as spectacular trans-oceanic dispersal events of Hypoxidaceae from Southern Africa to

Australia were not possible before 35 mya when West
Wind Drift was fully established. Furthermore, longdistance dispersal events of Astelia s.l. to geologically recent island archipelagos such as the Society or
Mascarenes Islands could not have occurred before emergence of these islands less than 4.5 mya.
The Plant List – a new global botanical resource
Hopper, S 1 , Raven, P 2
1 Royal Botanic Gardens, Kew, UK; 2 President Emeritus
Missouri Botanical Garden, St Louis, USA
The recent publication of the Plant List, a new widely accessible online working list of all plant species, represents the culmination of many years of effort on the part of a number of individuals and institutions. We review the major events and decisions that led to the delivery of what became known as Target 1 of the Global
Strategy for Plant Conservation and discuss lessons learned that might inform future approaches to challenges of this scale. We present some headline statistics from the Plant List and consider how they can be interpreted most appropriately. We conclude with a high-level survey of the opportunities presented by the availability of the list and its likely impact on the delivery of the GSPC over the next decade. Can the Plant
List provide a taxonomic background for the development of the world flora now sought by Target 1?
Does it provide an appropriate framework for the acceleration of progress towards Target 2? How should the availability of the list influence the development of the long-awaited GSPC toolkit?
Using heuristic methods to create the Plant List – approach and outcomes
Miller, C 1 , Allkin, R 2
R 1 , Nicholson, N 2
Lughada, E 2
, Govaerts, R
, Paige, J 1
2 , Hawkes, G 2
, Paton, A 2
, Magill,
, Welsby, J 2 , Nic
1 Missouri Botanical Garden, St Louis, USA;
Botanic Gardens, Kew, UK
2 Royal
Target 1 of the Global Strategy for Plant Conservation called for a working list of all known plant species by
2010. Although this target was carefully evaluated before adoption in 2002 and widely agreed to be feasible, by
2007 it was clear that, despite excellent progress, the
‘taxonomic impediment’ of a world-wide shortage of plant taxonomists made completion by 2010 unlikely and necessitated a creative approach to significantly accelerate delivery. In early 2008, scientists and IT specialists from Missouri Botanical Garden and the
Royal Botanic Gardens, Kew met to workshop possible alternative approaches to filling the delivery gap for
Target 1. Together they devised an innovative heuristic, rules-based approach to synthesize available regional and national floras, checklists and monographic data from data resources at Kew and the Tropicos® database at
MBG, merging them and resolving conflicts within and between the resources and used the resultant synthesized list to supplement available globally treated datasets like the World Checklist of Selected Plant Families at Kew,
Global Compositae Checklist and the International
Legume Database and Information Service in order to generate a working list of all species of angiosperms, gymnosperms, pteridophytes and bryophytes named The
Plant List which was both comprehensive and maximally informative with respect to synonymy. We present the assumptions on which this approach was founded and the algorithms applied to merge and resolve the data. We characterize the resulting resource, identifying its strengths and limitations. Where possible, we explore whether the limitations of The Plant List derive from the nature of the source data or from the approach adopted.
We conclude with an overview of enhancements to data inputs, algorithms and processes which have the potential to improve the quality of future releases of The Plant
List.
The significance of completing GSPC Target 1: what does it mean for the user?
Allkin, B 1 , Nic Lughadha, E 1
1 Royal Botanic Gardens, Kew, UK
The Royal Botanic Gardens Kew and Missouri Botanical
Garden have been working together to deliver GSPC
Target 1: a working global list of plant species. To achieve this by 2010, we sought to compare the various monographic and regional data resources held by our own institutions and collaborators and to derive from them a single list based on the taxonomic and nomenclatural judgements contained. We detected and resolved conflicts automatically; the algorithms and procedures used to generate ‘The Plant List’ are described elsewhere in this symposium. This paper presents a preliminary analysis of the resulting data set describing its strengths and weaknesses from a user perspective. We explore which of the quality criteria considered to define success are of most relevance to the user experience. That the botanical community has created a complete and synonymised list of the world’s plants for the first time has significance beyond systematic botany. The roles of different kinds of plant name lists and the audiences for them are examined. We illustrate the impact, in diverse disciplines, of the lack of easy access to a comprehensive and authoritative list. We present evidence of unnecessary and ongoing costs currently faced by organizations that build third-party information services to deliver other types of information about plants. Finally we discuss the role that The Plant
List might play in such services in the future and the broader benefits to end-users of those services.
The African Plant Database: integrating data from different sources
Chatelain, C 1 , Gautier, L 1
1
GF 3 , Phillipson, PB 2
, Callmander, MW 2
, Dobignard, A 1
, Smith,
, Loizeau, P-A 1
Conservatoire et Jardin Botaniques de la Ville de
Genève, Switzerland;
Louis, USA; 3
2 Missouri Botanical Garden, St
South African National Botanical Institute,
South Africa
311

The African Plant Database is accessible on the internet and presents information on taxonomy, nomenclature, distribution and ecology of all African flowering plants.
It originated from a collaboration between Conservatoire et Jardin botaniques de la Ville de Genève and South
African Botanical Institute and was limited to Tropical and Southern Africa. Through a collaboration with Alain
Dobignard and Tela Botanica, a second release has integrated species from North Africa. A third release includes Madagascan species through collaboration with
Missouri Botanical Garden. The database is updated on a regular basis to integrate publications of taxonomic treatments and new species. Merging data of different origins was of course not straightforward. Part of discrepancies were treated by application of standards
(spelling, authors). It was not our purpose to resolve discrepancies originating from different taxonomic views. However, we had to handle these descrepancies respectfully of rules of nomenclature. This enforced us to improve the structure of the database by integrating a hierarchical approach of names, including exhaustive synonym links. This database has proved to be of great help to many users and has been choosen as a nomenclatural backbone for the African Plant Initiative of the Andrew W. Mellon Foundation.
The Global Compositae Checklist: lessons in marrying biodiversity informatics with systematics
Flann, C 1 , Richards, K 2 , Wilton, A 2
1 Netherlands Centre for Biodiversity Naturalis (section
NHN), Wageningen University, The Netherlands;
2 Landcare Research, Manaaki Whenua, Lincoln, New
Zealand
The burgeoning field of biodiversity informatics is having a large impact on the manner of compiling taxonomic checklists. The Global Compositae Checklist utilises the possibilities of computer assisted checklist generation by integrating existing electronic taxonomic datasets from around the world for this large plant family
(more than 25000 species). Purpose built software (C-
INT) retains all original data and links names that are deemed the same to a 'consensus' name which reflects a summary of all contributed data. Insights from this process will be presented, discussing the advantages and pitfalls of trying to meld taxonomy and nomenclature with informatics technology. As well as this, contribution towards Target 1 of the Global Strategy for Plant
Conservation (GSPC) and The Plant List will be assessed. Issues that have arisen range from cross disciplinary expectations between two fields that have different focuses and priorities to differences in communication (e.g. computer languages versus that of botanical nomenclature). Reuse of software tools and code from other projects has proven technically more difficult than expected, despite the willingness to share.
Major issues have been encountered regarding obtaining and standardising data primarily due to the plethora of formats represented in the data contributed. Progress in obtaining global coverage will be reported. The intricacies of matching names will be discussed, accounting for the vast range of variations in spelling of epithets, author abbreviations, orthographic variants, typographic errors and electronic artifacts. The role of standards, particularly those of the Taxonomic Databases
312
Working Group (TDWG) will be touched upon.
Development of an approach to accommodating conflicting taxonomies will be outlined. Unexpected logical inconsistencies in data content will be reviewed and expectations that taxonomic experts will contribute voluntary vetting input will be examined. The questions of whether aggregation is enough and whether this process leads to fulfilling Target 1 with sufficient data content quality will be considered. The ultimate result of marrying biodiversity informatics with systematics is significant progress. Computers offer impressive advantages while the role of taxonomic and biodiversity informatics experts, in particular the relationship between them, remain integral to the success of these projects.
The Plant List: the contribution from liverworts and hornworts – a model and community-driven system
Von Konrat, M
M 3 , Engel, J 1 da Costa, D 7
1 , Söderström, L 2 , Hagborg, A
, Gradstein, R 4
, ZhU, R-L 8
, Pócs, T 5 , Váa, J 6
1 , Renner,
, Pinheiro
1 The Field Museum, Chicago, USA; 2 Norwegian
3
University of Science and Technology, Norway;
National Herbarium of New South Wales, Sydney,
Australia;
France;
4 Muséum National d'Histoire Naturelle, Paris,
5 Eszterházy College, Eger, Hungary;
7
6 Charles
8
University, Prague, Czech Republic; Instituto de
Pesquisas Jardim Botanico do Rio de Janeiro, Brazil;
East China Normal University, Shangai, China
The critical biological, environmental, and evolutionary significance of liverworts (Marchantiophyta) and hornworts (Anthocerophyta) is now well documented.
There remains a critical need to synthesize the vast amount of nomenclatural, taxonomic and global distributional data for these two groups of plants. These are fundamental components of developing a working list of all known plant species under the auspices of the
Convention on Biological Diversity (CBD). Such a synthesis has far reaching implications and applications, including providing an invaluable tool to taxonomists and phylogenetic systematists for analyzing phytogeographic and diversity patterns, aiding in the assessment of floristic and taxonomic knowledge, and identifying knowledge gaps in our understanding of the distribution and composition of liverwort and hornwort floras at a global scale. We outline a major internationally coordinated effort that is synthesizing the vast amount of nomenclatural, taxonomic and global distributional data for these unique groups of land plants.
The project includes over 20 active participants representing more than a dozen countries and 25 institutions. Prior to this effort there was no central source that attempted to harness core data from the immensely scattered literature. There are three fundamental components to the project: (1) a worldwide checklist of liverworts and hornworts that will ensure high quality data for The Plant List and other international databasing and bioinformatic initiatives; (2) regional or local distribution checklists; and (3) the synthesis of the taxonomy, systematics, and nomenclature for each taxonomic group. The project has been an active data provider to IPNI, Tropicos and CoL as well as numerous bryologists worldwide, and it has been generously supported by GBIF and EOL. The success of the project relies on strong collaboration

among institutions, international databasing initiatives, and the bryological community.
Phylogenomics of the seed plants
Cibrián-Jaramillo, A 1 , Little, DP 2
Martienssen, RA
S 5 ,
3 , Coruzzi, G 4
Stevenson, DW 2
, McCombie, WR
, DeSalle, R 1
3
, Wong, GK-
,
1 Sackler Institute for Comparative Genomics, American
Museum of Natural History, New York, USA; 2 Cullman
Program In Molecular Systematics, New York Botanical
Garden, Bronx, USA;
USA;
3 Cold Spring Harbor Laboratory,
4 Center for Genomics and Systems Biology, Dept of Biology, New York University, USA; 5 Dept of
Biological Sciences, University of Alberta, USA
We explore the genome-level influence of plant diversification using 32,833 sets of orthologs from the nuclear genomes of 185 plant species in 150 genera, analyzed in a combined analysis. The topology for the seed plants consists of the angiosperms as sister to a monophyletic gymnosperms with Gnetales sister to the rest of the living gymnosperms. Of particular focus is the origin of the integument and aril in the gymnosperms and the subsequent diversification of the seed and associated structures. Transcriptome data from ovule primordia and maturing ovules across the gymnosperms and selected angiosperms have been collected in order to elucidate the origin of the integument in seed plants and to try to determine which integument of the two in the angiosperms is the original integument found in gymnosperms. Additionally, the origin of the aril in the
Taxaceae has been investigated. Similarly, leaf primordia and developing leaves have been sampled in order to focus on the origin(s) of the diversification of gymnosperm leaves. We have used a novel phylogenomic approach to identify overrepresented functional gene categories at major nodes in our tree, revealing critical genes important to seed plant and gymnosperm diversification. For example, the second largest subunit of RNA polymerase IV and V (NRPD2) played a prominent role in the divergence of gymnosperms, consistent with maternal control of small
RNA in the seeds of flowering plants and the emergence of double fertilization in angiosperms.
Phytochrome data resolve the backbone of cycad phylogeny and reveal the recent, simultaneous diversification of large genera
Rai, HS 1 , Nagalingum, N 2, 6 ,
Stevenson, DW 3
, Little, DP
, Graham, SW 5
3 , Clements, MD 4
, Mathews, S 4
1 Utah State University, USA;
Berkeley, USA; 3
2 University of California,
4
New York Botanical Garden
Harvard University, USA; 5 University of British
Columbia, Canada; 6 Present address: National
Herbarium of NSW, Sydney, Australia
As part of the collaborative Gymnosperms on the Tree of
Life project, we present phylogenetic analyses of three independently evolving nuclear loci (PHYN, PHYO and
PHYP; and their associated introns) for one of the oldest crown group of seed plants, the cycads (Cycadales). The living cycads are represented by approximately 300 living species, of which we have sequenced approximately 200 species for one target nuclear gene
(PHYP). A subset of these taxa (24 species) were also included in a larger core-taxon data set that included sequences from all three target genes. Relationships among genera of the Cycadales have been ambiguous, primarily due to disagreements regarding the relative placement of a few genera along the backbone of the tree. Despite these disagreements, previous analyses of both molecular and morphological data have provided evidence that Cycas is sister to all other cycads, and have consistently united Zamia with Microcycas . Our combined nuclear data set contains a relatively small number of nuclear characters (6435 aligned nucleotides) that provides similar (and sometimes better) support to phylogenies produced from data sets that include many more characters (many of these analyses were based on genes/regions from organellar genomes). Our data strongly support (1) the position of Cycas as the sister to the remaining cycads, (2) a clade of Lepidozamia plus
Encephalartos , with Macrozamia as their sister, (3) a clade in which Zamia + Microcycas and Stangeria +
Ceratozamia are sister clades, with Bowenia as their sister, and (4) a clade of Dioon that is moderately supported as the sister group of all other Zamiaceae. The positions of Lepidozamia, Encephalartos, Macrozamia, and Ceratozamia are consistent with trees from previous analyses of morphological data, and with some, but not all, published molecular trees, but here we provide strong support for the individual (and separate) placement of
Bowenia and Stangeria (previously recognized as a monophyletic family, Stangeriaceae). Preliminary phylogenetic informativeness profiles (PIP) suggest that our nuclear data set may have greater phylogenetic power
(than previously sampled chloroplast regions) when applied to the extremely short branch lengths found within this ancient group of seed plants. In addition to these findings, molecular age estimates using our more inclusive single-gene data suggest that the short internode branch lengths present in the cycads may be due to a much more recent species radiation, a stark contrast to the ancient estimated age of the family (at least a Paleozoic origin). The strong support for the backbone of Cycadales and interfamilial relationships combined with our age estimates significantly advances our understanding of phylogenetic relationships within this ancient family, and provides a much stronger basis with which to recircumscribe the now non-monophyletic
Zamiaceae.
Integration of plastid phylogenomics, morphology and the fossil record to assess biogeographic patterns and divergence in Gnetales – the importance of sampling deep and broad
Ickert-Bond, S
Aguilera, J 3
1 , Rydin, C 2
, Metzgar, J 1
, Raubeson, L 3 , Peery, R 4 ,
2
1 University of Alaska Fairbanks, Dept of Biology and
Wildlife and UA Museum, Fairbanks, Alaska, USA;
Stockholm University, Dept of Botany, Stockholm,
Sweden; 3 Central Washington State University, Dept of
Biological Sciences, Ellensburg, USA; 4 University of
313

Illinois at Urbana-Champaign, Dept of Plant Biology,
USA
The Gnetales are a unique clade comprising very morphologically disparate genera such as the tropical liana Gnetum L. (31 species), the monotypic genus
Welwitschia Hook. (resembling 'stranded octopuses') from the Namib Desert and the desert shrub Ephedra L.
(45–50 species). However, the monophyly of these three genera is strongly supported based on previous molecular and morphological data sets. Comprehensive sampling of modern diversity is imperative for understanding evolutionary patterns in Gnetales, which are confounded by numerous incidences of parallelisms, intraspecific variation, and polyploidy. Furthermore, rate heterogeneity is particularly prominent in the Gnetales for all molecular data sets examined so far. Here we present results from multiplex sequencing-by-synthesis
(MSBS) of multiple plastid genomes on the Illumina
Genome Analyzer to resolve phylogenetic relationships in Ephedra and Gnetum and the Gnetales as a whole.
First, we PCR-amplified the plastid genomes using 25 primer pairs. Tagged microreads from each species were then assembled with a combination of de novo (YASRA) and reference-guided (RGA) assembly using previously published plastid genomes as references. We have amplified and obtained sequences using this method for over 30 gnetalean species (from Gnetum and Ephedra ) and we are using a Welwitschia plastid genome produced by shotgun sequencing. Based on combined evidence from plastid genome data, morphology and examination of early Cretaceous fossils we conclude that extant species diversity in Gnetales are the result of recent radiations in Ephedra and Gnetum that followed massive extinction of the Mesozoic diversity in the Oligocene and
Miocene. Biogeographic diversification in the Gnetales is complex including Beringian disjunctions and long distance dispersal aided by birds and lizards in Ephedra , as well as water dispersal in sea water in Gnetum .
Differentiation and diversification in the
Araucariaceae
Ruhsam, M 1
T 4 , Lowe, A 2
Thomas, P 3
, Biffin, E 2 , Ennos, R 3
, Matthews, S 5
, Gardner, M 1
, Mei, W 6 , Mill, R 1
, Jaffré,
, Rai, H 7 ,
1 Royal Botanic Garden Edinburgh, UK; 2 Australian
Centre for Evolutionary Biology and Biodiversity,
Adelaide, Australia; 3 University of Edinburgh, UK;
4 Institut de Recherche pour le Développement, Noumea,
New Caledonia;
Gainesville, USA; 7
5 Arnold Arboretum of Harvard
University, Cambridge MA, USA; 6 University of Florida,
Utah State University, Logan, USA
The Araucariaceae is an ancient family of gymnosperms which had its highest diversity in the Jurassic and
Cretaceous period when it occurred almost worldwide.
The extant members of the family have a predominantly
Southern Hemisphere distribution and consist of three genera Araucaria (19 species), Agathis (21 species) and
Wollemia (1 species). In this presentation we will summarise the results of recent research on the phylogeny of Araucariaceae and will then focus on the genus Araucaria . Araucaria (monkey puzzle trees) occurs in South America, Australia, New Caledonia,
Norfolk Island and New Guinea. Its centre of diversity is
314 in New Caledonia, where 13 of the world’s 19 species are endemic. To understand the evolutionary history of the genus, we present a dated phylogeny estimating the timescales of divergence of the different main clades, and discuss the uncertainties associated with these dates. We will also assess whether completely sequenced chloroplast genomes can provide insights into the patterns of diversification in the closely related assemblage of species in New Caledonia. Finally, to contribute towards our understanding of how so many wind pollinated species have diversified on such a small island (the main island of New Caledonia is ca. 410 km x
60 km) we will summarise our investigations into the spatial scales and landscape conditions over which populations become differentiated within individual species.
Progress in integrating molecular and morphological data in seed-plant phylogeny
Mathews, S
David 4 ,
D 7 , Lam, V 8
1 , Campbell, C
Graham, S
, Li, J 9
5
H, Raubeson, L, Rothwell, G 17
2
, Holman,
, Liston, A
, Cronn, R
2
10 , Little, D
Mei, W 12 , Morris, J, Nagalingum, N 14
3
11
, Gernandt,
, Ickert-Bond, S 6 , Kelch,
, Mapes, G,
, Parks, M 10 , Rai,
1
3
Harvard University, USA;
USDA Forest Service, USA;
Autonoma de Mexico;
Canada; 6
5
2 University of Maine, USA;
California, Berkeley, USA;
4
University of British Columbia,
University of Alaska, USA;
8
Universidad Nacional
7 University of
University of British
9
Columbia, Canada
11
Hope College, USA; 10 Oregon State University, USA;
New York Botanical Garden, USA;
Florida, Gainesville, USA;
12 University of
13 University of Texas,
Brownsville, USA; 14 University of California, Berkeley,
USA; present address: National Herbarium of NSW,
Sydney, Australia;
16
15 Utah State University, Logan, USA;
Central Washington University, USA; 17 Ohio
University, USA
Living gymnosperms comprise approximately 1000 species distributed in five major lineages: cycads,
Gingko , gnetophytes, Pinaceae, and cupressophytes. The extinct gymnosperms are distributed in at least 18 lineages with comparable morphological diversity: including Elkinsia , Lyginopteris , Quaestora , medullosans, gigantoperids, cordaites, Voltziales, bennettitites, glossopterids, corystosperms, peltasperms,
Caytoniales, Pentoxylales, Autunia , Laceya , and
Bilignea . We have now sampled nucleotide characters from nearly all living species: we recovered whole plastid genomes and multiple nuclear loci from about
16% of the living species, which we chose to cover critical nodes in currently available trees. These core taxa were also the focus of thorough morphological study here, along with parallel investigations of many of the extinct gymnosperm clades, integrating these different lines of evidence is challenging, but in meeting these challenges we aim for a synthetic understanding of seedplant evolution. Our studies have clarified major relationships within most of the extant gymnosperm clades, providing new insights into character evolution and clade diversification dates in each lineage. The persistent question of relationships among the major living seed plant lineages, and their extinct relatives is also addressed using these considerable new data.

Evolution of Compositae on islands: pattern and process
Baldwin, BG 1
1 University of California, Berkeley, USA
Members of Compositae constitute some of the most diverse plant lineages on islands worldwide. Recent phylogenetic studies in Compositae have revealed particularly striking examples of long-distance dispersal events involved in origins of insular lineages and of anatomical shifts on islands from ancestral herbaceousness to woodiness (and, rarely, vice versa).
Some diverse clades of different archipelagos thought to be only distantly related are now recognized as closely related (e.g. in Sonchinae); others long known to be closely related have been recently shown to have undergone parallel evolutionary changes upon colonization of different archipelagos (e.g. in Madiinae).
Those examples underscore the long-standing question of whether particular lineages have high intrinsic potential for success in insular environments, and, if so, why?
Recent documentation of pseudo-self-incompatibility
(SI) in various groups of island Compositae helps to reconcile Baker's Rule with success of some ancestrally self-incompatible insular lineages, as noted by Crawford and colleagues. Molecular-based age estimates and diversification rates for Compositae clades have reinforced and extended earlier conclusions about the youth of island radiations, especially in the Hawaiian islands, where the endemic Compositae were once viewed as ancient relicts. Hypotheses of homoploid hybrid speciation in island Compositae have gained additional support from recent studies in the Hawaiian and Macaronesian archipelagos, and other hybridization hypotheses await rigorous testing. In situ polyploidization remains a rare phenomenon in island
Compositae (and other island angiosperms), although ancestral polyploidy continues to be implicated in success of some (especially Pacific) lineages. Lack of molecular variation in DNA sequences commonly used for phylogenetic analyses has continued to be a major limitation for resolving evolutionary histories of island clades. Genomic-level sequence acquisition and analysis will likely lead to rapid progress in evolutionary understanding of insular taxa, including the resolution of cryptic diversity that is currently unrecognized and unprotected. Such progress is urgent in the face of rapid demise of island ecosystems from combined effects of invasive organisms and other anthropogenic environmental disruption.
Gene trees and species trees: examples from across the Compositae
Gemeinholzer, B 1 , Enke, N 2
1
2
University Giessen - Special Botany Dept, Germany;
Botanic Garden and Botanical Museum Berlin-Dahlem
(FU-Berlin), Germany
In many groups of the Compositae gene trees not evidently reflect the phylogenetic species affiliation of the organism under investigation. Discrepancies between signals of chloroplast and nuclear as well as morphological characters have been stated by several authors in many investigations of different lineages, e.g. within the Cardueae, Cichorieae, and Anthemideae and are a common phenomena within the Compositae.
Possible scenarios to explain this evolutionary pattern are either due to fast radiations in early stages of lineage developments and subsequent incomplete lineage sorting effects or hybridization events across lineages and signal persistence, called reticulate evolution. But also nuclear genes sometimes not evidently reflect species relationships, when structural changes of chromosomes, especially in polyploid or dysploid species, are results of hybridizing parents and subsequent agamospermy or incomplete lineage sorting. The complex patterns of bifurcating phylogenetic structures among taxa overlain by reticulate relationships have non-trivial implications for the reconstruction of lineage affiliation. Problematic hereby in all molecular investigations is the weak resolution of basal lineage affiliations, as transitional characters indicative of basal relationships are often scarce. Repeated rapid radiations and diversification must have taken place, in all different stages of
Compositae lineage development, which resulted in the large diversification of the family today.
Gene flow, hybridization and adaptation in wild,
1 domesticated, and weedy sunflowers
Kane, N 1 , King, M 1 , Rieseberg, L 1 , Andrew, R
University of British Columbia, Canada
1
Using EST sequence data from across the range of three wild species of Helianthus , as well as domesticated, weedy and invasive genotypes of H. annuus , we compare diversity, gene flow and divergence within and between species. Selective sweeps and adaptive introgression underline the importance of selection in enhancing gene flow at some loci, while disruptive or directional selection increases divergence at other loci.
Phylogeny, biogeography, and evolution of
Anaphalis
(Compositae: Gnaphalieae) in the eastern Himalayas
Nie, Z 1,2 , Sun, H 1 , Meng, Y 3 , Wen, J 2 , Funk, V 2
1 Kunming Institute of Botany, Chinese Academy of
Sciences, Kunming, China; 2 National Museum of Natural
3
History, Smithsonian Institution, Washington DC, USA;
Institute of Tibetan Plateau Research at Kunming, CAS,
China
The daisy tribe Gnaphalieae (Compositae) is well represented in the Southern Hemisphere, especially in southern Africa and western Australasia, with much lower taxonomic diversity in the Northern Hemisphere.
Anaphalis is the largest genus in the Gnaphalieae from
Asia with 80–110 species, mostly in Asia with one in
North America. The eastern Himalayas to Tibet Plateau of SW China harbor the greatest diversity of Anaphalis .
Existing molecular phylogenies have sampled very few taxa from Asia and it can be argued that Asia is the most important ‘missing’ part of the puzzle to construct the
315

evolutionary history of this tribe. The taxa so far sampled from Asia are positioned between the African basal grade and the western Australasian clades and are therefore critical for understanding the evolutionary transitions in the tribe. The internal and external transcribed spacers of the nuclear rDNA (ITS and ETS) were sequenced for
Anaphalis species, focusing on the eastern Himalayan taxa, in order to check the hypothesized monophyly of the Anaphalis and its diversification pattern in the eastern
Himalayan areas. The monophyly of the Anaphalis is weakly supported on the basis of current sampling and data. The genus is nested within the African–
Mediterranean Helichrysum and shows an affinity to
Mediterranean taxa. It seems that Anaphalis , together with European Helichrysum , has a common South
African origin. The species relationships within
Anaphalis are not well resolved but two groups of species are well supported and show a distinct biogeographic pattern that may indicate that the extant distribution of Anaphalis in the eastern Himalayan region resulted from at least two independent dispersals and/or radiations.
Understanding evolution in large clades: novel solutions to global problems in the largest flowering plant family on earth
Funk, VA 1 , Flann, C 2 , Bonifacino, JM 3
1 Smithsonian Institution, USA;
The Netherlands; 3
2 Wageningen University,
Universidad de la República, Uruguay
Examining evolution in large clades poses special challenges. There are new ways to identify problems, create solutions, and pool data to answer questions.
Under the umbrella of TICA (The International
Compositae Alliance) three projects are underway: the
Global Compositae Checklist (GCC; agreed upon names), the Virtual Key for the Compositae (VKComp; key to species), and Life Desks (fed into the
Encyclopedia of Life) with a possible link to WebTol.
All this will provide rapid availability of the latest information and greatly speed up science while making this large family accessible to conservation biologists and the general public. The Global Compositae Checklist underpins all of our efforts. First regional lists were combined along with complete species lists of some super generic taxa. These data were prepared and loaded into new software developed by Landcare Systems. It provides all of the variations of names and allows one to select the preferred name. A preliminary checklist (80% complete) is available to experts. The Virtual Key to the
Compositae began in 2009 and is an international effort to build an electronic key to identify any species in the family. This effort began with Phase I: North America.
There are three main issues: (a) creating a list of characters, (b) standardization of terminology, and )c) scoring the matrix. Some keys are already available on line. The Encyclopedia of Life (Lifedesks) is the result of a consortium of institutions that are working to make reliable information on organisms that inhabit Earth available to everyone. 'Lifedesks' allows 'curators' to build WebPages in a simple fashion before they are released to the public. TICA is working to promote the use of Lifedesks as a way of organizing and sharing information on taxa as it becomes available. Organizing the community is done by TICA. A 2010 workshop,
316 funded by the Synthesis Center of EOL, recommended that TICA create 'tribal coordinators' and 'regional coordinators' for the family. These coordinators are responsible for identifying team members and communicating with them on a regular basis. Answering scientific questions is the ultimate goal and we can use the products of these digital efforts to study questions about systematics, biogeography, and evolution. Imagine the power of knowing the correct name for each of the
25,000+ species in the family, their distribution, and their position on the phylogeny. Go further and imagine that this complex and fascinating family is made accessible to everyone by means of a virtual key and an EOL page, continuously updated. We could learn the origin of floras, which clades are the result of adaptive radiations, reconstruct ancestral groups, determine the accuracy of various hypotheses about the origin of the family, and much more. If we work together, these coming years are destined to be wonderful times for students of the
Compositae.
Hybridization and polyploidy in the evolution of the
1
Compositae: a case study in
Calendula
Plume, O 1
Cornell University, New York, USA
An estimated 70 percent of (if not all) angiosperms are of polyploid origin, and many are also hybrids. Both hybridization and polyploidy have genomic consequences which may include genome rearrangements, gene silencing and/or activation, and changes in location and timing of gene expression. These changes often generate morphological, biochemical, and reproductive diversity, some of which may become subject to selection and drive speciation. A few systems in the Compositae (e.g., Helianthus, Senecio,
Tragopogon ) have been at the forefront of the study of these processes and their evolutionary consequences.
Now, investigation of more lineages of Compositae is required to better characterize both small- and large-scale patterns of evolution in the family and is fundamental to understanding the impact of hybridization and polyploidy on the evolution of diversity across the entire family. The small Mediterranean genus Calendula includes 12 currently recognized species, the best known of which is
C. officinalis (pot marigold), cultivated for centuries for both medicinal and ornamental use. The genus is remarkable for the wide range of chromosome numbers reported for different species (2n=14, 18, 30, 32, 44, and
~85). These numbers have been hypothesized to be the result of hybridization events between species with different chromosome numbers followed by genome duplication (allopolyploidy). In order to elucidate relationships in the genus and to begin testing hypotheses of polyploid parentage, total genomic DNA was extracted from over 70 wild-collected or cultivated individuals representing all 12 species of Calendula , 12 of 15 subspecies of C. incana and C. suffruticosa , and 19 accessions from eight countries of the highly polymorphic and widely distributed C. arvensis , as well as from seven outgroup species in four genera of the
Calenduleae. Three non-coding chloroplast regions
(atpIatpH, petLpsbE, and ndhFrpl32), the internal transcribed spacers 1 and 2 of nuclear ribosomal DNA
(ITS), and two low-copy nuclear markers (CHS and A39)

were amplified, sequenced, and cloned (when necessary).
Separate parsimony analyses of the chloroplast and ITS datasets resolved the taxa into two major clades, a clade of four diploid species endemic to Morocco and a clade of all other taxa, and suggested extensive diversification of the plastid genome since divergence of these clades.
ITS copies appear to have homogenized toward those of the maternal progenitors and there is a loss of evidence of biparental inheritance of nrDNA. These datasets also suggested multiple origins of polyploid species with 44 and ~85 chromosomes. The low-copy nuclear markers tell a more complicated story of extensive hybridization and may help uncover the multiple ways in which the diploid genomes have combined to give rise to the remarkable diversity of species at various ploidy levels.
Eucalyptus
from the early Eocene of Patagonia,
Argentina: phylogenetic, biogeographic, and ecological implications for understanding eucalypt evolution
Hermsen, EJ 1 , Gandolfo, MA 1
, Nixon, KC 1
, Johnson, KR 2 , Zamaloa,
1
MC 3 , Wilf, P 4 , Cúneo, NR
Cornell University, Ithaca, USA;
5
University Park, USA; 5
2 Denver Museum of
Nature and Science, Denver, USA;
Buenos Aires, Argentina; 4
3 Universidad de
Pennsylvania State University,
Museo Paleontológico Egidio
Feruglio-CONICET, Trelew, Argentina
The early Eocene Laguna del Hunco beds of Patagonia,
Argentina contain a diverse fossil assemblage of angiosperms, gymnosperms, ferns, insects, fish, and frogs. 40Ar/39Ar radioisotopic analyses of tuffs interbedded with the fossiliferous, caldera-lake sediments indicate that the assemblage is ca. 51.9 Ma. Some of the most significant fossils from this site are abundant leaves, several buds, a flower, and numerous infructescences and isolated capsules with features characteristic of the extant genus Eucalyptus . The leaves are falcate with numerous, closely-spaced secondary veins and oil glands. Each bud displays a transverse scar separating the hypanthium from an apical operculum.
The scar is interpreted as having been left by loss of a caducous calyx, while the operculum is interpreted as corolline. The margins of the individual petals comprising the corolline operculum are visible, indicating that the petals were coherent rather than connate. The single specimen representing an open flower lacks a perianth, has a single style and stigma, and likely had numerous stamens (only some of which are preserved). Infructescences are composed of several umbellasters with 3–7 capsules each; capsules display valvate dehiscence and have perianth and staminophore scars as well as a prominent disc. Phylogenetic analyses indicate that the fossils belong within the Eucalyptus clade. The presence of a calycine scar on the flower buds is characteristic of species within and closely allied to
Eucalyptus subgenus Symphyomyrtus , indicating that the fossils have affinities to this subgenus. These fossils are currently the oldest record of Eucalyptus in the world, as previously the only reliably dated and diagnosed
Eucalyptus fossils were from Neogene sediments. The fossils also represent the only known natural occurrence of the genus outside Australasia. Given the derived phylogenetic position of the fossils, however, they do not provide direct evidence that Eucalyptus arose or underwent early diversification in South America. They instead provide the first reliable evidence for a more complicated, partially extra-Australasian scenario for the biogeographic history of the genus. The presence of
Eucalyptus at Laguna del Hunco is part of a broader pattern indicating biogeographic links between Eocene floras of Patagonia and modern floras of Australasia exemplified by other taxa from the flora, such as Akania
(Akaniaceae), Gymnostoma (Casuarinaceae), and
Papuacedrus (Cupressaceae). Seedlings of modern
Eucalyptus subgenus Symphyomyrtus that occur in caldera habitats of Papua New Guinea today make use of volcanic destruction of standing rainforest to establish themselves, thus suggesting a possible ecological analog for Eucalyptus at Laguna del Hunco.
Functional and evolutionary relationships between eucalypt terpene synthases
Keszei, A 1 , Padovan, A 1 , Carter, R 1 , Foley, W 1
1 Research School of Biology, Australian National
University, Canberra, Australia
Eucalypts represent the largest genus of trees in
Australia, and are also one of the chemically most variable groups of plants. Their terpene-dominated leaf oils have been shown to influence herbivory, and have also been implicated as a response to climatic stress.
Despite an abundance of chemical data, very little work has focussed on the biochemistry of terpene production, and its relevance as an adaptive strategy which may have shaped the genus through evolution. Based on terpene synthase transcripts from 19 species, we compare phylogenetic relationships, gene function and leaf oil phenotypes to shed light on the evolutionary processes leading to the chemical diversity observed in the genus today. The expressed terpene synthases from eucalypts fall into two well- defined monophyletic groups, one containing the sesquiterpene (C
15 other the monoterpene (C
10
) synthases and the
) synthases. The trees based on DNA sequence comparisons show well supported groups which do not reflect the phylogenetic histories of the species. Comparison of leaf chemistries and gene function of the expressed genesindicates that the best supported clades represent functional separation predating the cladogenesis of Eucalyptus . We also describe foliar terpene compositions and transcriptomes of chemical mosaics and individuals from natural populations to gain an insight into the mechanisms behind the diversity of terpenes in Eucalyptus , and how this can be used in reconstructing the evolutionary history, and predicting the evolutionary potential of chemical variability in the genus.
Reconstructing phylogeny of the eastern Australian clade of monocalypts (
Eucalyptus subgenus
Eucalyptus
)
Bayly, M 1 , Ladiges, P 1 , Nelson, G 1
317

1 School of Botany, The University of Melbourne,
Australia
Eucalyptus subgenus Eucalyptus (commonly called the
‘monocalypts’) is strongly supported as a monophyletic group, with flowers characterised by a single petaline operculum, lack of free sepals, and anatropous ovules arranged in two rows per loculus. This second-largest subgenus, with at least 106 species, includes some of
Australia’s most important timber trees, such as E. marginata (jarrah) and E. regnans (mountain ash), the latter the tallest flowering plant in the world. The group is assumed to date from the early Palaeogene and distributed largely in the wetter areas of both southwestern and eastern Australia. The eastern species form a significant clade, which ranges from tropical to cool temperate latitudes, but is most common in southern coastal and upland regions. Among the eastern clade, subgroups are traditionally recognised by morphology and differentiated in terms of habitat and ecology; they include northern mahoganies, stringybarks, peppermints, snow gums, scribbly gums, blue and green ashes. Despite the distinctiveness of each of these eastern subgroups, morphological synapomorphies relating them to one another are few and DNA sequence variation not high for determining phylogenetic relationships based on any one data set. A meta-analysis combining all available data is presented to hypothesise relationships and biogeographic patterns in eastern Australia.
The role of hybridisation in the evolution of eucalypts in south-eastern Australia
Jones, RC 1 , McKinnon, GE 1
1
BM 2
, Vaillancourt, RE 2 , Potts,
School of Plant Science, University of Tasmania,
Australia; 2 Cooperative Research Centre for Forestry,
Tasmania, Australia
Many plant genera exhibit geographically structured sharing of cytoplasmic DNA markers among species, consistent with the hypothesis of hybridisation, introgression and chloroplast capture rather than shared ancestral polymorphism. Eucalyptus is a large and taxonomically complex genus with over 700 species dominating Australia’s forests and woodlands. Our studies of maternally inherited chloroplast (cp) DNA in this genus suggests that many eucalypt species persisted in glacial refugia in south-eastern Tasmania where introgressive hybridisation and chloroplast capture occurred among multiple species, involving more species than reported in American and European tree genera.
Apart from extensive sharing of the major chloroplast lineages amongst species in this region, our main evidence for such hybridisation comes from a study of co-occurring populations of Tasmanian blue gum
( Eucalyptus globulus ) and heart-leafed silver gum ( E. cordata ). E. cordata is a rare endemic of south-eastern
Tasmania, which co-occurs with the more widespread E. globulus . These species are morphologically divergent and do not form a hybrid zone. Despite this, in a broadscale study of mixed populations of the two species, we found evidence that E. globulus has acquired cpDNA from E. cordata in multiple populations. Further studies have revealed that this cpDNA exchange has been accompanied by nuclear (nr) DNA exchange. While E.
318 globulus and E. cordata retain strongly differentiated nuclear gene pools overall, introgressed nuclear anonymous (Amplified Fragment Length
Polymorphisms, AFLP) and mapped (Diversity Array
Technology, DArT) markers from E. cordata have been incorporated into the E. globulus gene pool, with some markers introgressed repeatedly in more than one population. At the finer scale, the frequency of E. cordata nrDNA markers in E. globulus decreased with distance from E. cordata . This gene exchange has potential effects on genetic diversity and adaptation in E. globulus and other Tasmanian eucalypts, as well as the organisms dependent on these species. The recent sequencing of the eucalypt genome allows us to identify the genomic regions that are exchanged compared with those that maintain species integrity in the face of introgression.
Comparative phylogeography of three south-east
Australian
Eucalyptus
species with different ecological tolerances: signals of shared responses to past climate change and chloroplast introgression during recolonisation
Nevill, PG 1,2 , Ades, PK 3 , Bayly, MJ 4
1 Kings Park and Botanic Gardens, West Perth,
Australia;
Australia;
2
3
University of Western Australia, Nedlands,
School of Forest and Ecosystem Science,
University of Melbourne, Parkville, Australia;
Botany, University of Melbourne, Australia
4 School of
Past climate changes and associated range contraction, expansion and introgression events are regarded as major forces shaping the evolution of the genus Eucalyptus .
Previous investigations in south-eastern Australia have identified some putative refugia and recolonised areas for tall eucalypt forests during the last glacial cycle but the response of individual species are very little understood and in some regions, the evidence for migration or persistence is completely absent. We examined the phylogeography of three south-east Australian trees
( Eucalyptus delegatensis , E. obliqua , and E. regnans ) with different tolerances, in terms of cold, drought and soils, to explore whether: (1) species with different ecologies share major phylogeographic disjunctions; (2) sharing of cpDNA (introgression) is more likely at postulated recolonised locations than in glacial refugia.
Cambial tissue samples were collected from trees of E. delegatensis (n=120), E. regnans (n=270) and E. obliqua
(n=255) at 27 locations across south-eastern mainland
Australia and the large island of Tasmania. Trees were genotyped using five chloroplast DNA microsatellite markers. The number of haplotypes, number of unique haplotypes (the number of haplotypes found only in that species at that location) and haplotypes shared was calculated for each species at each location. For each species, mean within-population genetic diversity, total genetic diversity, and population differentiation were calculated treating alleles as unordered (hS, hT and GST) and ordered (vS, vT, and RST). To visualise relationships between haplotypes a statistical parsimony tree was constructed using the median joining network algorithm to construct the shortest, least complex tree. The species shared major genetic disjunctions (e.g. between
Tasmania, central Victoria and East Gippsland), and common patterns at proposed glacial refugia (generally

high haplotype diversity) and recolonised areas (low diversity). Inter-specific sharing of haplotypes was extensive, and fixation of shared, regional haplotypes was more likely in areas postulated as treeless at the Last
Glacial Maximum (LGM). Despite ecological differences, microsatellite data suggest the three species have responded to past climatic changes in a similar way, by persisting locally, in or near their current range, often in what would have been unfavorable conditions.
Therefore, rather than the waves of recolonisation or long distance dispersal experienced by tree species in much of
Europe or North America, E. delegatensis , E. obliqua and E. regnans , with the exception of the Central
Highlands region of Victoria in mainland Australia and the central region of Tasmania, do not appear to have migrated great distances. Although sharing of cpDNA occurred both in putative refugia and areas thought to be recolonised post-LGM, it seems likely that the process of recolonisation, in particular, results in loss of chloroplast diversity within species and interspecific haplotype sharing. The implications of these results for understanding forest responses to climate change and for current gene pool management will be discussed.
Genetic diversity and mating systems in eucalypt evolution
Byrne, M 1
1 Dept of Environment and Conservation, Australia
Evolutionary change at the level of species is fundamentally driven by processes occurring at the gene, genome and population level. Genetic diversity and selection drive the evolutionary processes of adaptation and speciation. Mutation, migration and random drift influence the level and pattern of genetic diversity and together with natural selection contribute to the cumulative change that underpins evolution. Population genetic studies in eucalypts reveal high levels of diversity that are generally maintained through a mixed mating system where outcrossing predominates but there is a significant fraction of inbreeding. The maintenance of outcrossed breeding systems in eucalypts capable of self pollination is facilitated by post-zygotic incompatibility mechanisms. Historical gene flow was high in most eucalypt species as genetic differentiation is low to moderate, except for species with significant range disjunctions. High estimates of the ratio of gene flow by pollen to seed demonstrate that gene flow is mainly a function of pollen dispersal as seed dispersal is quite limited. Pollen dispersal is generally mediated by nonspecific biotic vectors such as insects and birds. Long distance pollen dispersal, identified through paternity analysis, can be extensive, maintaining genetic connectivity at the landscape scale. Clonality in mallee eucalypts is relatively common as they resprout from the lignotuber following disturbance. Clonality may exacerbate inbreeding in small populations and large infertile clones have been identified in some populations.
Phylogeny and evolution of the Polytrichopsida
, Hyvönen, J 2
2
1
Bell, N 1, 2
Botanical Museum, University of Helsinki, Finland;
Plant Biology, University of Helsinki, Finland
The Polytrichopsida occupy a critical position in the bryophyte tree of life as the only major moss lineage outside of the Bryopsida having significant levels of both species diversity and macro-morphological variation.
They include the largest of all extant bryophytes with the most fully developed vascular systems and are often characterised by a three-dimensionally complex leaf structure that can be interpreted as a pseudo-mesophyll.
A unique and structurally complex spore dispersal apparatus has evolved within the class as a parallel development to the arthrodontous peristomes found in the Bryopsida. If extant bryophytes are the representatives of the oldest embryophyte lineages, now largely sidelined by evolution into ecologically subsidiary roles as epiphytes and other diminutive denizens of the boundary layer, then the Polytrichopsida may offer us the best insights into how bryophytes may once have been (or had the potential to be) more dominant components of terrestrial ecosystems. Our recent work has provided the first robust and fully developed phylogeny for the class, allowing major evolutionary trends to be polarised and identifying parallel evolutionary developments that have previously confounded generic-level taxonomy. While large and complex gametophytes appear to have emerged early in the history of the group, there have been several independent origins of lineages of small plants with reduced lamellae, many of them currently classified in
Oligotrichum . These parallelisms and others often appear to reflect symmetrical adaptation to similar ecological roles in the Northern and Southern Hemispheres, with the class as a whole showing strong phytogeographic patterning. The well-known and predominantly Northern
Hemisphere genera Polytrichum and Polytrichastrum are shown to have been unnaturally circumscribed in the past, with new sporophyte characters providing the basis for a classification that is congruent with the strong evidence from molecular data. Pogonatum represents a speciose radiation of gametophytic diversity based upon a highly distinctive but conserved sporophyte morphology, although its phylogenetic origins are ambiguous – incongruence between nuclear and organellar molecular data may be interpreted as supporting an ancient intergeneric hybrid origin for the genus. Previous generic-level taxonomy within the class has focused insufficiently on sporophytic variation, particularly of the peristome/epiphragm complex. We show that there is considerable diversity in this apparatus across the group, possibly related to functional adaptations towards efficient spore release.
Progress in inferring the deep phylogenetic relationships of mosses using plastid data
Chang, Y 1 , Graham, SW 1
319

1 Botanical Garden and Centre for Plant Research and
Dept of Botany, University of British Columbia, Canada
Recent studies have shed much light on relationships among the earliest branches of moss (Bryophyta) phylogeny, but there is still considerable uncertainty surrounding the relative arrangement of several major clades. Most previous large-scale bryophyte studies have applied taxon-intensive sampling using only one or a few genes. Here we retrieved a large number of slowlyevolving DNA markers (17 plastid genes and associated non-coding regions) from 48 exemplar species that are representative of the core groups of mosses and relatives.
We inferred moss backbone relationships in the context of land-plant evolution, using maximum likelihood (ML) and maximum parsimony (MP). Besides providing additional evidence for a subset of published findings, our results substantially improve support for branching patterns that were previously ambiguous in some analyses, including the very first splits in moss phylogeny. For example, we find substantial support for the following relationships: (i) Takakia as the sister group of all other mosses, followed by Sphagnum as the next successive sister group; (ii) Andreaea and
Andreaeobryum as sister taxa; (iii) Timmia (Timmiidae) as the sister group of a clade comprising Dicranidae and
Bryidae. We recovered multiple major branches within
Funariidae and Dicranidae, generally with strong support.
Relationships within Bryidae tended to be less well supported, as were the placements of Polytrichopsida and
Tetraphidopsida. We uncovered and investigated major conflicts among different analysis methods, and explored potential causes for these conflicts in a variety of ways, including the use of rate-filtered analyses. Discrepancies between MP and ML analyses are likely a consequence of long-branch artefacts. We also used simple perturbations to demonstrate that relatively minor differences in ML analysis conditions can lead to substantial conflicts, for some branches of moss phylogeny.
Phylogeny of haplolepideous mosses – challenges and perspectives
Stech, M 1
1 Netherlands Centre for Biodiversity Naturalis, Section
NHN, Leiden University, The Netherlands
The haplolepideous mosses (Dicranidae) form the second largest group of mosses and are morphologically and ecologically highly diverse. Early molecular phylogenetic reconstructions already provided support for the monophyly of Dicranidae and valuable insights into systematic relationships at the family level.
Examples are the positions of families with reduced peristomes (Amphidiaceae, Archidiaceae,
Catoscopiaceae, Ephemeraceae, Erpodiaceae) in the
Dicranidae and the re-circumscription of Dicranaceae, with several dicranoid genera having been transferred to
Leucobryaceae. Later, familial and generic relationships have been addressed based on more detailed analyses, for example, in Grimmiales, Calymperaceae, or in the largest moss family, Pottiaceae. However, major challenges remain at all taxonomic levels. No comprehensive multigene analyses have been performed after the initial molecular phylogenies to address the open questions of
320 relationships between the major haplolepideous lineages.
Examples are the highly polyphyletic Ditrichaceae and the status of ‘proto-haplolepideous’ species from different families, which were resolved as a basal grade in early phylogenies. Furthermore, Dicranidae include several of the most species-rich moss genera for which still only few molecular data are available (e.g. Fissidens ,
Dicranella, Leucobryum ). But even species-rich genera that have been studied in more detail, e.g. Campylopus , cope with a number of problems, such as extremely different variability of molecular markers and incongruence between molecular clades and morphological species circumscriptions. This talk will provide an overview of the current state and most urgent remaining problems in unravelling relationships in the haplolepideous mosses. Characteristic features of the evolution of molecular markers as well as of morphological and ecological traits will be discussed as a basis and perspective for future (molecular) approaches on relationships and character evolution in the
Dicranidae.
1
Recent advances in the phylogeny of diplolepideousalternate mosses
Quandt, D 1 , Noben, S 1 , Bell, N 2 , Stech, M 3 , Newton, A 4
Nees Institute for Biodiversity of Plants, University of
Bonn, Germany; 2 University of Helsinki, Finland;
3 Nationaal Herbarium Nederland, Universiteit Leiden
Branch, The Netherlands
Despite the increasing ease of generating molecular data for phylogenetic systematics we still often face the
'unresolved backbone' problem, i.e. ambiguous or unsupported relationships between clearly monophyletic lineages. The most stunning bryophyte example perhaps comes from the apical pleurocarp clade, as until now no molecular phylogenetic approach has been able to provide a robust hypothesis of relationships among the
Hypnales. In addition, it is apparent that likelihood and
Bayesian approaches at times propose topologies that conflict with maximum parsimony. For example, under parsimony the Bartramiales/Hedwigiales comprise the sister clade to the pleurocarpids, i.e. the pleurocarpous and acrocarpous Orthodontiales, Rhizogoniales,
Aulacomniales together with the Hypnodendrales,
Ptychomniales, Hypnales and Hookeriales, whereas model-based approaches favour the Orthotrichales as sister to the pleurocarpids clade with maximal support.
Thus the branching order among the early diverging lineages in the by far largest moss clade that harbours
70% of species diversity is still unknown, as is the lineage giving rise to pleurocarpy which resulted in the major moss radiation. Here we will summarize the current state of knowledge concerning the diplolepideous-alternate moss phylogeny and reveal some surprising systematic changes, such as the resolved position of the enigmatic genus Pulchrinodus , the
Catoscopiaceae and the Helicophyllaceae. Based on sequence data from four plastid ( rbc L, rpl 16, rps 4, trn L-
F) and one mitochondrial marker (nad5) we provide a new hypothesis of the branching order among early diverging diplolepideous-alternate mosses and discuss the phylogenetic utility of the markers employed for bryophyte phylogenetics.

Evolution of gametophytic versus sporophytic characters in the moss order Hookeriales (Bryophyta)
Pokorny, CI 1 , Chuan Ho, B 1 , Frahm, J-P 1
Shaw, J 1
1 Duke University, USA
, Quandt, D 1 ,
Convergent evolution and reversibility hamper reconstructing evolutionary histories. The haploid gametophyte is the dominant generation in bryophytes.
The sporophyte is the diploid generation where meiosis takes place and is partially dependent on the gametophyte. These two alternating generations experience different selective pressures, which can lead to opposing evolutionary trends that can further complicate our understanding of their evolutionary histories. Incongruence between classifications emphasizing gametophyte versus sporophyte characters is well exemplified by the pleurocarpous moss order
Hookeriales. Although the branching order among early diverging pleurocarps (Hypnodendrales first,
Ptychomniales second) is fairly well established, relationships within the crown group (Hypnales and
Hookeriales) remain challenging, apparently because of extensive homoplasy in morphological traits. 122 species representing 71 genera of pleurocarpous mosses were sampled for five DNA regions in the plastid ( trn L-F & rps 4), mitochondrial ( nad 5) and the nuclear (ITS & 26S) genomes. Five species from the Hypnodendrales and
Ptychomniales were included as outgroups. The ingroup consists of 95 species of Hookeriales and 22 species of
Hypnales. Six morphological characters (four gametophytic and two sporophytic), often used in classifications of this order, and three types of habitat occupation, were scored for ancestral state reconstructions on the resulting phylogeny, and to identify the occurrence and frequency of reversals in morphological characters. Character state reconstructions and correlated evolution of morphological traits and habitat occupation was performed in BayesTraits. Our analyses show that reversals have occurred in five of the six morphological characters reconstructed. Strict epiphytism was the only type of habitat with sufficient phylogenetic signal that could be unambiguously reconstructed on our phylogenetic tree. We also found correlations among morphological characters, between as well as within generations, and with one type of habitat occupation (strict epiphytism), though no causation could be established for these correlations.
Disentangling the knots of rapid evolution: origin and diversification of the moss order Hypnales
1
Huttunen, S 1 , Pleurocarps Evolutionary Research
Group, PERG 2
Turku, Finland;
2
University of http://www.pleurocarps.eu/
The moss order Hypnales includes ca. 40 % of all mosses and with around 4500 species it belongs to one of the largest lineages among early land plants. Despite significant research efforts, the phylogeny and familial level relationships are still unresolved which hampers testing large scale hypothesis on diversification and phylogeograpy or morphological radiation of the group.
We compiled a data matrix including 141 taxa and four markers from three genomes: nuclear ITS1-5.8S-ITS2, mitochondrial nad 5 and plastid encoded rps 4 and trn L-F.
As our main interest was to study diversification and relationships especially at family level and below, our sampling included one to four species from the most
Hypnelian moss families. Despite of a mixture of rapidly evolving as well as more conserved markers deeper nodes in the phylogeny remained relatively poorly supported and it was not possible to reliably indentify, for example, the first evolving lineages within Hypnales.
In addition the already documented sister group relationship of the Hypnales and Hookeriales could not be inferred with confidence. At family level groupings of the species and genera were very similar to earlier analyses and suggest that circumscriptions of many families are still in need for re-evaluation. The failure to provide a robust phylogeny of the pleurocarpous mosses with the standard sequencing approaches is now well documented and indicates that unless a joined effort will be undertaken using multi-gene analyses via next generation sequencing techniques we will not be able to address the rapid radiation at early stages of the hypnalean evolution.
From Vienna to Melbourne via Latin America: what became of the African Plants Initiative?
Crane, PR 1
1 Yale School of Forestry & Environmental Studies, USA
The African Plants Initiative (API), introduced at the
XVIIth AETFAT conference in Addis Ababa, Ethiopia, in 2003, had as its primary aim the development of a high quality and sustainable database of images of
African plants based mainly on herbarium specimens. It also sought to digitize, and to make available in electronic form, other kinds of resources relating to
African plants. To guide this work and also to ensure a comprehensive approach, the API also helped accelerate the development of a synonymized checklist of African plants. The success of the African Plants Initiative made it possible to develop a similar approach with partners in
Latin America, and this effort is now well under way.
Progress with the Latin American Plants Initiative
(LAPI) now raises the prospect of developing similar efforts in Asia and Australia, and these are now being actively pursued. The possibility of developing a global on-line database of images of the world's plants, with all the benefits that would bring, is now a realistic possibility. One important question for the immediate future is the extent to which this collaboration can be extended to include new partners, and new kinds of partners. Another issue is how best to develop and use this new resource to support teaching as well as research.
In the long term a further key question is the extent to which it will be desirable, or even possible, for even the largest plant diversity institutions in the world, to develop independently and sustain the significant ongoing investments in hardware, software and computer
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support that will be needed to curate and make available electronic images of potentially millions of specimens.
An alternative model, to outsource such activities to a specialist not-for-profit group, is one potential solution that needs to be given serious consideration.
Who are our users and what do they want from us?
Gallagher, Michael 1
1 JSTOR Plant Science
Just as the African Plants Initiative has flourished and grown over the years into the Global Plants Initiative, the online interface to the images and data created in these projects has been continually updated and refined and now resides as part of JSTOR Plant Science
(http://plants.jstor.org/). Digitized materials include herbarium specimens, current and historical scanned literature, photographs of living plants, drawings, prints, correspondence and more. Does this wealth of information meet the needs of our users, and how well have we designed an interface to suite them? How has our user base changed over time? Has it expanded as more resource types have been added? Do any of these changes correlate with the addition of social networking features? What would it take to interest more users outside of science and academia – expanded content types, less technical metadata, a different type of navigation? What content have the users found most interesting? What impact does online access to this information have on our users in terms of time and effort saved, costs, and quality of work produced? Are users primarily searching for specific content about species and geography or do they browse through various resource types? Do the search and browse interfaces easily get our users exactly what they need? Here we look at what the user statistics, feedback and workshops tell us about how
JSTOR Plant Science is being used and how it is shaping future development.
Use of JSTOR Plant Science as a fundamental botanical diversity resource
Davis, A 1 , Darbyshire, I 1 , Briggs, M 1
1 Herbarium, Library, Art and Archives (HLAA)
The JSTOR Plant Science website (http://plants.jstor.org) provides quick and easy access to (herbarium) type specimens, and other original material, produced by the
African Plants Initiative (API), Latin American Plants
Initiative (LAPI) and Global Plants Initiative (GPI). The original aim of these projects was to provide a sustainable digital library of scholarly resources about the plants of Africa, Latin America and, subsequently, the world. Realizing the potential of these resources, botanists from different institutes have independently produced their own ‘virtual floras/herbaria’ through the compilation of downloaded information from JSTOR and the World Checklist of Selected Plant Families
(http://apps.kew.org/wcsp/), using generic software. The applications of these ‘virtual floras’ are many and wide ranging, but the most common use by far is the identification of specimens. There are several key attributes of these hand-made identification tools, including: accuracy; speed (virtual searches are often
322 quicker than using keys or engaging in herbarium searches); coverage (more types are available than in any one herbarium); specimen care (due to a reduction in specimen handling). Use of these ‘virtual floras’ for identification of Rubiaceae specimens has shown that the taxonomic coverage achieved by API, for example, is exceedingly high. It has also been observed that curators are keen to add the types of new taxa, and type specimens overlooked (e.g. during the API project), to
JSTOR via GPI, to further improve taxonomic coverage and as part of best-practice curation (e.g. to enable online access of herbarium data and to reduce specimen handling). The completion of GPI will be a significant achievement and of considerable value for a wideranging audience. For those using GPI-derived JSTOR
Plant Science to produce their own ‘virtual floras’ and identification tools, the next logical step is perceived to be a virtual world flora, through the linking a global species checklist to JSTOR Plant Science, and other electronic resources.
Global Plants Initiative – possible future directions
1
Saw, LG 1
Forest Research Institute, Malaysia
Digital access to taxonomic material in the last ten years has progressed very rapidly. The basic material for plant taxonomic research, access to type specimens and literature have now gone digital and become available in the internet. Projects such as the Global Plants Initiative
(GPI) and the JSTOR Plant Science and Biodiversity
Heritage Library have benefited many developing countries by making such material freely available.
Although there is no substitution to examining actual specimens in taxonomic revisions, such efforts will certainly facilitate projects in particular for many tropical countries in South-east Asia where such material are not readily available but are often held in European and US herbaria. The Tree Flora of Sabah and Sarawak and the
Flora of Peninsular Malaysia projects have tapped into these resources, in particular, type specimens from the
National Herbarium of the Netherlands at Leiden and more recently from the Singapore Botanic Gardens. One very important spin off in the digitising of herbarium specimens is the use of the data for conservation research. The capture of locality information from herbarium specimens has become a powerful tool to document plant distribution and provide the basis for plant conservation assessments. In Malaysia, for example, such herbarium based assessments coupled with field surveys has jump started plant conservation efforts. Analysis of such data is now being used to identify centres of plant diversity and plant endemism, providing quantitative data to identify the most important plant areas for conservation. At a global scale such data is used to understand plant geography and more recently in plant and vegetation responses to climate change and glacial maximums of recent ice ages. From the perspective of users of taxonomic data and resources,
GPI could look into the following:
• There is a need to have Global Plants Initiative (GPI) and the JSTOR Plant Science to have a more inclusive policy, currently there is difficulty for non-collaborating institutions assessing the data.

• Improving and keeping identification standards by engaging authorities and linking to regional and national flora projects and experts.
• Many herbaria are moving towards providing access to their collections in virtual herbarium and many national projects have started to database their collections, GPI could collaborate with such projects and link these databases thus reducing duplication of data entry.
• Extending GPI to Asia in a more focused manner, i.e. prioritising regions that are most taxonomically active, countries with active national flora projects.
• Initiate geo-referencing of data set for conservation and biogeographic research. The database of specimens could extend beyond types, may include national endemic taxa.
There is a need to link with national institutions and local botanists who are more familiar with the gazetteer information to ensure better precision in geo-referencing the data.
Curating a virtual herbarium – new challenges for collection management
Chesselet, P 1 , Labat, J-N† 1 , Pignal, M 1 , Haevermans, T 1
1 Muséum National d'Histoire Naturelle, Paris, France
The Global Plants Initiative (GPI), a collaborative project among more than 150 institutes from all continents, aims to create a sustainable digital library of scholarly resources about the world’s plants, with at its core, high resolution images of nomenclatural type specimens. For
MNHN Paris, participation in GPI and its predecessors
(African Plants Initiative and Latin American Plants
Initiative) has been important for the valorization of its collections and the discovery of types, thus highlighting the immense wealth of the herbarium in both numbers of specimens and coverage, as well as in its long history of plant collecting. GPI has accelerated access to the unique resources that we curate for the world’s scientists but has also increased our understanding of their global significance and raised their profile nationally and internationally. Here we explore how the development and expansion of specimen data capture and high resolution imaging have impacted the role of the herbarium collections manager. It goes without saying that certain tasks that were hitherto difficult, if not impossible, are greatly facilitated by the availability of digital records. However, the requirement to curate a digital collection necessitates a novel approach that must be recognized and appropriately resourced. At MNHN, the digitization initiatives designed to contribute to
JSTOR Plant Science have laid the foundation for a further, nationally funded industrial specimen digitization endeavour on a scale not previously attempted by any of the world’s major herbaria. We describe the scope and scale of this pioneering project and report on progress after the first year.
JSTOR Plant Science: the past, present and future
Guthrie, K 1
1 JSTOR Plant Science
JSTOR Plant Science ( http://plants.jstor.org
) is an online environment that brings together content, tools, and people interested in plant science. It provides access to foundational content vital to plant science – plant type specimens, taxonomic information, scientific literature, and related materials, making them widely accessible to the plant science community as well as to researchers in other fields and to the public. It also provides an easy to use interface with powerful functionality that supports research and teaching, including the ability to measure and record plant specimens, share observations and objects with colleagues and classmates, and investigate global plant biodiversity. JSTOR Plant Science strives to be a comprehensive online research tool for aggregating and exploring the world's botanical resources, thereby dramatically improving access for students, scholars, and scientists around the globe. It is useful for those researching, teaching or studying botany, biology, ecology, environmental and conservation studies. The majority of the content available on JSTOR Plant Science has been contributed through an effort known as the
Global Plants Initiative (GPI). GPI is an international undertaking by leading herbaria to digitize and make available plant type specimens and other holdings used by botanists and others working in plant science every day. Partners include more than 170 institutions in 60 countries. There are two partner networks in place and contributing today: the African Plants Initiative which focuses on plants from Africa and the Latin American
Plants Initiative which contributes plants from Latin
America. GPI is also expanding to Asia with a first partner working from Nepal. GPI has received funding and guidance from the Andrew W. Mellon Foundation.
Without these partners, JSTOR Plant Science would not be possible. Our partners play a central role in selecting and providing content, advising us, setting priorities, obtaining permissions, capturing meta-data, preparing contextual materials, and providing feedback on how best to present the materials online. The content and the tools that comprise JSTOR Plant Science are driven by the plant science community, including leading experts around the world. How can the momentum of this collaboration be maintained over the coming years?
What additional collaborations might this network of international institutions produce? At the next IBC in
2017, what should JSTOR Plant Science look like? How will it be different? How well do we understand our users and are planned developments in line with their current and future needs? Are there new communities of users for the GPI resource that were not anticipated at the outset of the project?
Relationships of African Genistoid legumes
Boatwright, JS 1 , van Wyk, B-E 2 , le Roux, M 2 , Wink,
1
M 3
South African National Biodiversity Institute, Compton
Herbarium, Cape Town, South Africa; Dept of Botany and Plant Biotechnology, University of Johannesburg,
3
2
South Africa; Institute of Pharmacy and Molecular
Biotechnology, University of Heidelberg, Germany
The 'core Genistoids' are an alliance of tribes in the
Fabaceae which comprises the Crotalarieae, Euchresteae,
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Genisteae, Podalyrieae, Sophoreae (in part) and
Thermopsideae. These taxa, centered in Africa and
Eurasia, all share a similar alkaloid chemistry, notably the presence of quinolizidine or more rarely pyrrolizidine alkaloids. Much progress has been made, especially over the last two decades, to clarify intertribal and intergeneric relationships in this group, with a particular focus on the
African Genistoid legumes. These are contained within the tribes Crotalarieae, Genisteae, Podalyrieae and
Sophoreae. Studies of morphological, chemical, cytological, anatomical and molecular data in combination have resulted in important changes at the generic level, contributing towards a more natural and phylogenetic generic classification system for the
Genistoids. This lecture is aimed at reviewing these important changes to the Genistoid classification and the data leading to the resultant re-circumscriptions of genera. The enigmatic genus Cadia , infamous for its pink, actinomorphic flowers, was shown to be closely related to the Podalyrieae and transferred to this tribe from Sophoreae, based on morphological, chemical, cytological and molecular data, increasing the number of genera from eight to nine. Sophora inhambanensis , which shares similar alkaloids and seed flavonoids with the Podalyrieae was suspected of being closely related to this tribe. Through molecular data it was shown, however, to be more closely related to the other Sophora species, suggesting rather that these chemical characters are the result of convergence. The most substantial changes occurred within the Crotalarieae, resulting in an increase of the number of genera from 11 to 16. This has emanated from the generic reclassification of Lebeckia s.l. and Lotononis s.l. Both these genera were found to be polyphyletic. Lebeckia was divided into three genera –
Lebeckia s.s., a reinstated Calobota and a new genus,
Wiborgiella (with nine species). The monotypic North
African genus Spartidium was included in Calobota .
Lotononis was divided into five genera – Lotononis s.s., the reinstated Listia , Leobordea and Euchlora , as well as a new monotypic genus, Ezoloba . Morphological, chemical, cytological, anatomical and molecular evidence contributed towards these changes. Studies of character evolution have revealed many examples of convergent evolution for several characters, which complicate generic circumscription considerably.
A first molecular phylogeny for the genus
Crotalaria
(Fabaceae, Crotalarieae)
Le Roux, MM 1 , Boatwright, JS 2
Moteetee, AN 1
, Van Wyk, B-E 1 ,
1 University of Johannesburg, South Africa; 2 South
African National Biodiversity Institute, Cape Town,
South Africa
Crotalaria L. comprises some 702 species distributed across the Southern Hemisphere and extending into Asia and North America. Tropical Africa (including
Madagascar) is the main centre of diversity for the genus, with 540 species occurring in this region, followed by
India (92 spp.) and Brazil (45 spp.). The genus is characterised by a combination of diagnostic characters which include a rostrate keel, highly inflated pods, strongly dimorphic 5+5 anther arrangement, hairy style and the presence of macrocyclic pyrrolizidine alkaloids.
Polhill established an infrageneric classification system
324 that was later modified as a result of taximetric analyses by Bisby and Polhill. Ansari recently completed a taxonomic revision of the Indian species and further modified the infrageneric classification. Crotalaria is one of the last remaining genera in the tribe Crotalarieae for which molecular studies have yet to be done to explore infrageneric relationships. A first molecular phylogeny for the genus will be presented based on parsimony and
Bayesian analyses of nrITS and psbA-trnH data. The morphological patterns, on which the current classification system is based, are largely congruent with the molecular results. However, the molecular results in many cases revealed a more natural infrageneric placement for some of the species and groups of species
(the latter often geographically correlated).
1
Phylogeny of sections
Alburnoides, Spartopsis and
Verzinum
of
Cytisus
L. (Fabaceae)
Auvray, G 1 , Malécot, V 2
INRA Angers-Nantes,
Beaucouzé/Angers, France; 2
UMR GenHort,
Agrocampus-Ouest, Centre d'Angers, France
Within the controversial genus Cytisus L. (Fabaceae), the sections Alburnoides , Spartopsis and Verzinum , are at the origin of the diversity of colors found in broom cultivars.
These three Mediterranean and North African sections seem to share a close proximity both morphologically and phylogenetically. However, only few phylogenetic studies have focused on Cytisus or parts of the genus and relationships between and within the three sections remain unclear. Based on a more exhaustive taxonsampling of sections Alburnoides , Spartopsis and
Verzinum , the present work aims to clarify those relationships. Two chloroplastic intergenic spacers ( trn D-
T and trn S-G) and two nuclear markers (ITS and
LEGCYC) have been sequenced for 43 accessions representing the fifteen taxa recognized in the three sections as well as five taxa used as outgroups. Both parsimony and Bayesian analyses have been performed on independent and combined data sets. Phylogenetic trees reveal the monophyly of each section. Section
Alburnoides seems to be the sister-group of section
Verzinum . Both those sections cluster in a clade shown as the sister-group of section Spartopsis . Relationships within sections are not well resolved. Only two clades appear within section Spartopsis , separating Cytisus striatus , Cytisus scoparius and its subspecies from
Cytisus cantabricus . This phylogenetic study confirms the delimitation of the three sections revealed by a recent taxonomic study based on statistics analyses of floral characters. Both morphological and molecular analyses have revealed the difficulty in discriminating species within those three sections.
Phylogeny of the tribe Indigofereae (Leguminosae –
Papilionoideae): geographically structured more in succulent-rich and temperate settings than in grassrich environments
Schire, BD 1 , Lavin, M 1 , Barker, NP 1 , Forest, F 1
1 Royal Botanic Gardens Kew, UK

The Papilionoiod genus Indigofera is the third largest in the legumes. Molecular studies have revealed that the genus comprises four lineages, and that the radiation of the genus as a whole began approximately 16 Myr ago.
One of the four lineages (termed the Cape Clade) is restricted to the south-western (mainly winter-rainfall) region of southern Africa (although early divergent elements have an arid affinity to southern Namibia and the Richtersveld). This clade will form the focus of this presentation. We present an updated and expanded phylogeny for the Cape clade, and investigate the factors that might have driven diversification in this group. The link between phylogeny and areas of endemism is explored, and we discuss phytogeographic connections and distribution patterns, We also present preliminary data on the role of edaphic factors as a possible cause of diversification in this clade.
A review of African Phaseoleae
Moteetee, A 1 , van Wyk, B-E 1
1 University of Johannesburg, South Africa
This presentation gives a broad overview of the tribe
Phaseoleae in Africa. The tribe is distributed throughout the tropics, subtropics and warm-temperate regions of the world. It is recognizable by the generally twining nature of the stems, leaflets subtended by stipels and inflorescences that are pseudoracemes. Phaseoleae is traditionally divided into seven subtribes, but recent molecular data has shown that the tribe is both polyphyletic and paraphyletic, as a result the placement of some subtribes (and some genera) is uncertain. Based on these molecular analyses, there has been a suggestion that Phaseoleae sensu latu should be recognized to include the following: four of its current subtribes
(Kennedinae, Cajaninae, Phaseolinae and Glycininae), the tribes Desmodieae and Psoraleeae (at subtribal level) and an assortment of basally branching genera including
Decorsea , Otoptera and Psophocarpus (previously in
Phaseolinae), Erythrina and Mucuna (previously
Erythrininae). However, as currently circumscribed
Phaseoleae comprises 89 genera (making it the largest tribe of the subfamily Papilionoideae in number of genera) and approximately 1580 species. In Sub-Saharan
Africa the tribe is represented by 480 species distributed in 43 genera, including 14 endemic ( Adenodolichos,
Alistilus, Bolusafra, Clitoriopsis, Dipogon, Eminia,
Neorautanenia, Nesphostylis, Physostigma, Pseudenimia,
Pseudoeriosema, Pseudovigna, Sphenostylis, Wajira ) and three subendemic ones ( Decorsea, Otoptera,
Psophocarpus ). The pantropical genus Rhynchosia is by far the largest with 138 species, followed by Vigna with
64 species and Dolichos comprising 60 species. These three genera are mainly centred in Africa (and
Madagascar in the case of Rhynchosia ).
On the African Psoraleoid legumes (Fabaceae) – taxonomy, phylogenetic relationships and drivers of evolution
Stirton, CH 1
S 1
, Dludlu, M 1 , Muasya, AM
1 University of Cape Town, South Africa
1 , Chimphango,
147 species in 3 genera comprise the African Psoraleeae
(Fabaceae). We present molecular and morphological phylogenetic data that confirms the generic status and monophyly of African Otholobium, Psoralea , and
Cullen , and show that Hallia is nested within Psoralea .
The American representatives of Otholobium , transferred recently into the genus, are not congeneric with the
African species and represent a separate and potentially new genus. Otholobium and Psoralea have a centre of diversity in the winter rainfall area of Southern Africa, and originate at the onset of Mediterranean type climate in late Miocene. We present preliminary data that shows that seedling establishment, pollinator strategies, and soil nutrition are important evolutionary drivers in Psoralea and Otholobium .
Investigation of diversity and endemism of
Papilionoid legumes in Iran and cooperation with
1
ILDIS project
Ghahremaninejad, F 1
Tarbiat Moallem University, Iran
The research is concerned to the preparing of a perfect checklist for the Iranian Papilionoid legume (=Fabaceae:
Faboideae subfamily) taxa. This checklist is a part of a scientific work as titled 'Legumes of Iran, a checklist' and is a contribution to the ILDIS global project
(www.ildis.org). Iranian Papilionoid legume includes several tribes. The genera are nearly 60; these are including more than 1300 native species which less than half are endemic. The value of the genera is not the same for the flora of Iran. 22 genera include only one species, e.g. Caragana, Galega, Ebenus ; and 9 genera only with two species, e.g. Sesbania , Cullen ; but some other genera are so rich. These legumes are distributed nearly in all part of the country; but with a concentration on Iranian mountain areas (like Alborz Mountain ranges and
Zagross Mountain ranges). In this group Astragalus is the most important and diverse genus, with an endemism rate of more than 60%. Iran is one of the centers of genetic diversity for this huge genus. To the east the importance of this genus decreases. It includes more than 11 percent of the Iran’s flora, but for example it includes less than
1.3% of flora of China. After Astragalus the richest genera are Oxytropis, Trifolium, Vicia, Medicago, and
Hedysarum . After Astragalus , the richest endemic species are found in three genera: Onobrychis, Oxytropis and Hedysarum . Like Astragalus , these genera have high speciation rate in Iranian highlands. Iran can be one of the centers of diversity for these genera too. Members of some genera of Papilionoid legumes in Iran are introduced by man, with few species; thus these are not native (0.1% of the subfamily species). These cultivated taxa belong to Phaseolus, Vigna, Robinia, Lathyrus,
Trifolium, Medicago, Clitoria, Sophora, and Arachis .
The biggest part of the data is extracted from Flora
Iranica treatments; 'flora of Iran' treatments, and several published papers in diverse botanical journals. All of this data and some local unpublished investigations are used for making this checklist.
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Molecular phylogenetics of the tribe Hedysareae
(Fabaceae–Papilionoideae)
Kazempour Osaloo, S 1 , Amirahmadi, A 1 , Moein, F 1 ,
1
Kaveh, A 1
Dept of Plant Biology, Tarbiat Modares University,
Tehran, Iran
The tribe Hedysareae is one of the 28 tribes of Fabaceae–
Faboideae distributed mainly in Eurasia. A total of
103/89 accessions representing 90/76 ingroup taxa including 71/62 species of Hedysareae and 18/15 species of Astragalean and Caraganean clades (both of the tribe
Galegeae) plus 7 species of Vicioid clade and
Glycyrrhiza glabra as outgroups were included in phylogenetic analyses using nrDNA ITS and chloroplast trn L-F regions, respectively. The sequence data was obtained through PCR amplification of these fragments with appropriate primers and using the cycle sequencing reaction run in the 3730xl DNA Analyzer. The resulting sequence data was aligned using Muscle program.
Maximum Parsimony approach as implemented in
PAUP* using heuristic search strategy and Bayesian inference using MrBayes as well as Maximum
Likelihood method by GARLI were performed.
Phylogenetic analyses of single and combined datasets showed that all of the recognized Hedysaroid genera were nested in a single clade (Hedysaroid clade).
Astragalean and Caraganean clades were well allied with
Hedysaroid clade. Within this clade, Alhagi is the first branch which followed by Sulla species along with
Hedysarum humile and H. aculeolatum forming a well supported clade. Ebenus and Taverniera are each monophyletic and weakly sister to a large clade of
Hedysarum (including Sartoria hedysaroides ),
Onobrychis , Stracheya tibetica ( Hedysarum tibeticum ),
Eversmannia skkubspinosa and Corethrodendron scoparium . Hedysarum , at the current status, is not monophyletic. Forty-two species of Onobrychis plus E. subspinosa and C. scoparium formed a well supported clade. Onobrychis subgenus Onobrychis is paraphyletic but Onobrychis subgenus Sisyrosema is a well supported monophyletic group. PCR and DNA sequencing of the complete trn L-F region revealed that in the two arctic
Hedysarum species ( H. alpinum and H. boreale ), the trn LUAA intron, group I intron, was completely lost, which indicates that this rare genomic structural mutation was occurred before the divergence of these two species.
This is the first record for the loss of the trn LUAA intron in land plants.
Phylogenetic relationships within Coluteoid clade with special reference to the Eurasian genus
Colutea
(Fabaceae–Galeageae) based on nrDNA ITS and cpDNA mat
K sequences
Saadati, N 1 , Hosseini, H 1 , Kazempour Osaloo, S 1
1 Dept of Plant Biology, Tarbiat Modares University,
Tehran, Iran
The Coluteoid clade at the current status comprises
West/Central Asian Colutea , (including Oreophysa mirophylla ), Sphaerophysa , Smirnowia , Eremosparton and Podlechiella , East Asian Phyllolobium , South
African Lessertia and Sutherlandia , and Australian/New
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Zealander Swainsona , Carmichaelia and Clianthus .
Colutea is a medium-sized genus of ca. 30 species with shrubby habit and inflated pods distributed in the
Mediterranean region through west Asia to China. It has been divided into three/four sections. A total of 45 accessions representing 42 species including 12 Colutea species, 16 species of its allied genera, three Oxytropis species and nine Astragalus species plus 2 species of
Caragana and Chesneya as outgroups were included in phylogenetic analyses using both nrDNA ITS and cpDNA mat K sequences. Maximum parsimony,
Maximum Likelihood and Bayesian methods were used for phylogenetic reconstructions. Almost all analyses retrieved the well suppored Coluteoid clade, in which
Podlechiella vogelli forms the basalmost branch. The next branch is the Australian/New Zealand taxa followed by Phyllobium subclade as sister to a large assemblage of three highly supported subclades. One subclade composed of Lessertia (including Sutherlandia fructicosa ). The second subclade comprises Smirnowia turkestana – Eremosparton flaccidum plus Sphaerophysa .
And third one is Colutea in which species relationship is somewhat weakly resolved. Likewise, monophyly of none of its three multi-specific sections Colutea ,
Rostrata and Armata is not supported by the present molecular data. A specimen of Sphaerophysa salsula that has been previously misidentified as Astragalus cysticalyx , well allied with other S. salsula accessions; whereas a true specimen of A. cysticalyx analyzed in this study is a member of Astragalus s. str. clade.
1
Myrtaceae: progress on all fronts
Wilson, PG 1 , Heslewood, M 1
National Herbarium of New South Wales, Sydney,
Australia
Following the previous Myrtaceae symposium, held in
Melbourne in 2003, a new infrafamilial classification was published, based on mat K sequence data, which provided a new baseline for continuing research on the family.
This paper will outline progress in further resolving relationships and generic limits in the family, although a major emphasis will be on the sister taxa Leptospermeae and Chamelaucieae. The chloroplast regions mat K and the rbc L– atp B spacer, plus part of the External
Transcribed Spacer (ETS) of nrDNA, were sequenced. In
Leptospermeae, the major focus has been on testing the monophyly of the genus Leptospermum and examining its relationships to the other genera in the tribe. The genus, as presently circumscribed, is not monophyletic and resolves into two groups, one of which is paraphyletic with respect to the small genus Neofabricia .
In Chamelaucieae, work is proceeding on both morphological and molecular fronts to determine generic boundaries and unravel relationships amongst species.
Chamelaucieae is a diverse tribe of mostly shrubby plants, many with reduced flowers and small, indehiscent fruits. The small size of floral parts has contributed historically to the adoption of broad generic concepts that

are only now being resolved into monophyletic taxa. This paper will highlight several examples from this tribe and will also touch on particular difficulties encountered when considering conversion of the phylogeny into an acceptable taxonomy for some groups.
Sorting out
Syzygium
: progress and prospects for one of the most species rich of angiosperm genera
Biffin, E 1 , Craven, L 2
1 University of Adelaide, Australia; 2 Australian National
Herbarium and Centre for Biodiversity Research,
Canberra, Australia
With between 1200 and 1500 species, the myrtaceous genus Syzygium is one of the largest of angiosperm genera, and is a strongly significant component of humid forest ecosystems through much of the Palaeotropics.
Like other large and predominantly tropical lineages,
Syzygium has been plagued with taxonomic uncertainty, and this hinders documentation of this group’s diversity.
To date, there has been no coherent attempt to revise
Syzygium as a single entity. The majority of research has been locally based, with a focus on alpha taxonomy, and with little consistency, workers have variously emphasised sets of characters that can distinguish among species within a localised region. One important consequence has been confusion surrounding the definition of higher taxa within Syzygium . On the one hand, there has been a tendency to recognise a large and heterogeneous Syzygium along with various smaller segregate genera, while alternatively some workers have argued that an inclusive concept of Syzygium would best serve the group’s taxonomy. Here, we describe recent progress, and prospects, for syzygioid taxonomy and in particular, recent analyses of molecular phylogenetic data have provided insights into relationships that have substantially informed taxonomy. For instance, seed characters have been considered important in the delineation of higher taxa in Syzygium ; e.g. the Acmena and Syzygium sub-alliances are defined (respectively) by the presence, or otherwise of an intercotyledonary intrusion (ramifying tissues that are enclosed by and interlock the cotyledons). From recent molecular phylogenetic studies, there is little support for the recognition of groupings based upon seed morphology.
However, this conflict appears to be resolved by elucidating and invoking convergent evolution the seed.
In light of these new sources of evidence, we have argued for the recognition of a single large genus,
Syzygium . Significant areas of ongoing research need include the completion of floristic surveys for major regions that have been inadequately investigated, including Myanmar, Peninsula Malaysia, Kalimantan,
Sulawesi and the Philippines. Resolving interrelationships of subgenus Syzygium , which includes perhaps more than 90% of species in the genus, remains a challenge, although new and developing genetic approaches provide potential. We consider these areas significant not only from the biodiversity perspective.
For instance, Syzygium is a key group for interpreting wide-ranging evolutionary questions including regional biogeography, and as an exceptionally species rich lineage, understanding the processes that generate such broad disparities in amongst lineage diversities. We discuss these aspects in the context of molecular phylogeny.
Phylogenetic analysis of
Eugenia
(Myrtaceae), with emphasis on infrageneric circumscription and taxonomic implications
Mazine Capelo, F 1 , Castro Souza, V 2
E 3
, Forest, F 3 , Lucas,
2
1 Universidade Federal de São Carlos, Brazil;
Universidade de Sao Paulo, Brazil; 3 Royal Botanic
Gardens Kew, UK
Eugenia L. is an unwieldy and ecologically important genus of trees or shrubs with a pantropical distribution. It is the largest genus of Neotropical Myrtaceae, comprising c. 1000 species, distributed from Southern
Mexico, Cuba and the Antilles to Uruguay and
Argentina, with a small number of species (c. 60) in
Africa and an even smaller representation in Southeast
Asia and the Pacific. Eugenia species are remarkable in their number and in their morphological homogeneity that has resulted in taxonomic neglect and a terrible reputation as an ‘impossible’ group. Evolutionary relationships within Eugenia are unclear, there is no reliable way to divide the group into manageable groups for systematic study identification to species is a frustrating and routine problem. Species are regularly missed or mis-named in ecological inventories/surveys of the forests in which they are most diverse, causing serious problems for biodiversity management as well as preventing the use of Eugenia species as environmental tools. A phylogenetic hypothesis of Neotropical Eugenia, including associated genera and species from Africa and the Pacific was produced using DNA sequence data (ITS,
ETS and psb Atrn H). This analysis aimed to verify infrageneric groups of Eugenia as well as suites of supporting morphological characters, to determine relationships between groups and produce a framework for future taxonomic research. The resulting topology confirmed the inclusion of Neotropical Calycorectes,
Hexachlamys, Phyllocalyx and Stenocalyx in Eugenia to preserve a monophyletic Eugenia. Seven infra-generic groups can now be recognized within Eugenia. These are
Eugenia sect. Eugenia, Eugenia sect. Racemosae ,
Eugenia sect. Umbellatae , Eugenia sect. Phyllocalyx ,
Eugenia sect. Hexachlamys comb. ined., Eugenia sect.
Calycorectes comb. ined. and a new section to be published. This talk will present the results of the phylogenetic analysis in conjunction with supporting morphology, will discuss the possibilities and logic underlying decisions to classify Eugenia as proposed and will describe future challenges in implementing the classification, including typification.
Not so good on paper: evolution and phylogeography of the Australian Broadleaf Paperbark (
Melaleuca leucadendra
: Myrtaceae) complex
1
Edwards, R 1
University of Queensland, Australia
The Broadleaf Paperbark ( Melaleuca leucadendra ) species complex is a dominant component of Australia's tropical monsoonsoon flora. Comprising 14 poorly
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differentiated morphospecies, local scale ecological variation may play a role in segregating morphotypes in the face of more widespread geneflow. We examine species limits in a group where asymmetrical gene flow and a lack of clear differentiation between morphotypes in nuclear or chloroplast DNA suggests recent or ongoing hybridisation.
Psidium
phylogeny and evolution
1
Costa, IR 1 , Forni-Martins, ER 2
CEB 4 , Dornelas, MC 2 , Lucas, E 3
, Forest, F 3 , Proença,
Universidade Federal do Ceará, Brazil;
Estadual de Campinas, Brazil;
Kew, UK; 4
3
2 Universidade
Royal Botanic Gardens,
Universidade de Brasília, Brazil
With ca. 100 species, Psidium (Myrtaceae) is an important component of Neotropical flora and widespread in the Brazilian Cerrado and Tropical
Atlantic Rain Forest. This is a highly taxonomically complex genus and some natural hybridization cases have been identified. Recent phylogenetic studies indicate Psidium to be a monophyletic taxon, although relatively small taxanomic sampling leaves delimitation of some species unclear. To help assist taxonomic delimitation and understanding of species, in this work we applied different methodologies: (1) phylogenetic analysis based on molecular data, (2) chromosomal and
(3) genome size studies in an evolutionary context. The phylogenetic analysis used Parsimony and Bayesian
Inference of 26 species (24% of the generic diversity) and includes morphologically related genera such as
Acca, Accara, Campomanesia, Mosiera, Myrrhinium,
Pimenta and Ugni. The sampling comprised native species collected in the Amazonia, Brazilian Cerrado,
Caatinga and Tropical Atlantic Rain Forest biomes.
Myrrhinium appears sister to Psidium with the resulting cladogram indicating three principal clades and a clear biogeographical pattern. The species endemic to the
Cerrado vegetation appear in two separate clades, each with good statistical support: one emerges as sister group to a clade comprising species from Caatinga and Tropical
Atlantic Rain Forest, while the other cerrado clade is sister to Amazonian species. The chromosomal approach includes the majority of taxa sampled in the phylogenetic analysis: Acca (1), Accara (1), Campomanesia (9),
Myrrhinium (1), Mosiera (1), Pimenta (1) and Psidium
(11). All species present small chromosomes (<2µm).
The diploid chromosome number 2n=2x=22 occurred in these genera, exception for Psidium, where this number was observed in only two species (P. guajava and P. guineense). Polyploidy was observed in 82% of species of Psidium with a big variation on the ploidy levels
(2n=22, 44, 55, 66, 88). For P. cattleianum tetra
(2n=4x=44) and hexaploid (2n=6x=66) cytotypes were found, where 6x populations occurs at highest altitude and 4x were distributed in other Atlantic Tropical Rain
Forest localities with lower altitudes, next to the sea level. The distribution and number of rDNA 45S sites suggest a hybrid origin of the hexaploid. The genome size (2C-values) was determined by flow cytometry for
10 species of Psidium. The intra-specific variation of 2Cvalue was 9-fold, in a positive correlation with ploidy level. This approach reinforces the indications of an allopolyploid origin of 6x cytotypes of P. cattleianum.
Future studies will increase the sampling of molecular
328 and chromosomal data and will help clarify the species relationships and biogeographical patterns from Psidium, and explain the origins of polyploidy in these differential environments.
Extant and fossil Myrtaceae pollen morphology and its use in understanding family systematics,
1 phylogenetics and evolution
Thornhill, A 1 , Ho, S 2 , Macphail, M 3 , Crisp, M 1
Research School of Biology, Australian National
University, Canberra, Australia; 2 School of Biological
Sciences, University of Sydney, Australia; 3 College of
Asia and the Pacific, Australian National University,
Canberra, Australia
The identification and implementation of reliable fossil calibrations represents a key component of many molecular studies of evolutionary timescales. Although most paleontological calibrations are associated with macrofossils, the pollen record can also inform age calibrations if fossils matching extant pollen groups are found. Recent palynology work on over 200 taxa using
Scanning Electron Microscopy (SEM) and 400 taxa using Light Microscopy (LM) has shown that pollen of the myrtle family, Myrtaceae, can be classified into a number of characteristic groups that display phylogenetic signal on molecular trees. Records of Myrtaceidites
(Myrtaceae fossil pollen) extend back to as early as the
Paleocene and possibly Late Cretaceous. We measured the fit of 26 fossil pollen to the molecular phylogeny using parsimony optimisation of characters obtained from extant Myrtaceae pollen and identified seven
Myrtaceidites fossils appropriate for calibration. The age of these pollen, combined with ages of various
Myrtaceae macrofossils, were used to inform age constraints to calibrate a relaxed molecular clock in a
Bayesian phylogenetic analysis using molecular data comprised of two sequences from the chloroplast genome
(matK and ndhF) and one nuclear locus (ITS). The resulting trees can then be used to test Myrtaceae vicariance or dispersal event hypotheses.
1
The Banks and Solander collections – a benchmark for understanding the New Zealand flora
Brownsey, PJ 1
Museum of New Zealand, Wellington, New Zealand
New Zealand was one of the last significant landmasses to have been occupied by humans. Polynesians arrived about 750 years ago, and European colonisation was initiated following James Cook’s Endeavour voyage of
1769–1770. Human influence quickly resulted in massive changes to the country’s flora, including introductions of plants from many parts of the world. Just 240 years after
Cook’s voyage, the number of naturalised vascular plants

now slightly exceeds that of natives in a total flora of around 5000 species. Because New Zealand is an oceanic island archipelago occupied relatively late by humans, it is possible to characterise much of the flora as either native or naturalised, and to identify with reasonable certainty when, and from where, individual species were introduced. The collections made by Joseph Banks and
Daniel Solander on the Endeavour voyage are hugely important in providing a verifiable record of plants growing in New Zealand before European settlement.
They include species now assumed to be native, plus a few of tropical origin that were probably brought by
Polynesian migration. The collection of over 360 species therefore provides a vital benchmark for understanding the New Zealand flora. Cook’s circum-navigation of
New Zealand enabled Banks and Solander to botanise at several sites, particularly in the North Island and Cook
Strait. Most of the plants they collected were previously unknown to European botanists. They all returned to
England as Banks’s personal property, but were later acquired by what is now the Natural History Museum in
London. Solander studied them, coined new names and drafted descriptions for every species. Banks, meanwhile, hired a team of artists and engravers to convert the sketches made by Sydney Parkinson, artist on the
Endeavour voyage, into plates to illustrate the new species. Sadly, Banks’s grand plan for a Flora of New
Zealand never eventuated, and it was left to later botanists to publish formal descriptions of the new species, whilst the engravings only finally appeared in
Banks Florilegium in 1985. Ninety years earlier, in 1895, sets of about 300 duplicate specimens were returned to
New Zealand to assist in the production of a major botanical work, Thomas Kirk’s 'Students’ Flora of New
Zealand and the Outlying Islands'. Kirk realised that the number of introduced plants was so great that he needed the guidance of the Banks and Solander collections to distinguish native from naturalised. Prints from the original copper plate engravings were also sent out, and these, together with subsequent gifts of specimens, are now held in various institutions in New Zealand, including Te Papa. The Banks and Solander collections continue to influence taxonomic research on the New
Zealand flora. Even today they are cited in the descriptions of new species that are being recognised more than two centuries after their initial discovery. Most significantly, however, they are regularly admired by back-of-house Museum visitors as a tangible connection with Cook’s first voyage of exploration.
Botany Bay and the plants collected by Banks and
Solander: a benchmark for 18th century scientific collecting
Benson, D 1
1 Botanic Gardens Trust, Sydney, Australia
After observing the Transit of Venus at Tahiti and circumnavigating the islands of New Zealand, the
Endeavour turned westward and in April 1770 the east coast of Australia was sighted. The northern, western and southern coastlines of New Holland were already known as a result of Dutch exploration in the 17th Century but the east coast was unknown, as was the flora of the continent. The only pr record of Australian plants were some drawings and collections made by William
Dampier on the Western Australian coastline in 1699.
The events of first landfall and eight day stay at Botany
Bay (April 28–May 6) have traditionally focussed on
Kurnell, on the southern shore of the Bay. However a careful look of where some of the original plant specimens are likely to have been collected (using material held at the National Herbarium of New South
Wales in Sydney; now databased and maintained as a
Special Historical Collection) and at Cook, Banks and
Solanders’ day to day activities, shows that the visit was a carefully organised scientific exploration of the whole of the Bay and its surrounds. From the ship exploratory parties systematically radiated out, to maximise collecting opportunities. Collections were made, plant specimens were pressed and dried in paper folders, scientific descriptions written in the field, and drawings with field colours were done by the artists. Banks reports that 200 quires of papers were used and he himself spent one day looking after the drying material. The difference of the Botany Bay flora to anything previously encountered, and the number and variety of plant species amazed the botanists. Despite the fact that April–May is late in the flowering season we can establish that they collected at least 132 species at Botany Bay – this would be higher but many specimens collected in Australia were simply labelled New Holland and could have been collected at Botany Bay or in Queensland. Finally in the naming of Botany Bay it is evident that Cook could see how much the flora had impressed the botanists. The
Botany Bay visit appears to have been planned as the scientific recording site for the east coast. Subsequent stops on the Queensland coast were of much shorter duration except for the 6 weeks at Endeavour River which was unavoidable as a result of the damage to the ship. Banks’ favourable experiences at Botany Bay led to his recommending it as a site for a convict settlement, and to his ongoing support for botanists and collectors including Robert Brown to continue the description of the Australian flora.
1
The Virtual
Endeavour
Cafferty, S 1
Natural History Museum, London, UK
The Virtual Endeavour is a project based at the Natural
History Museum in London, dedicated to the collections of Banks & Solander made on the famous Endeavour voyage. The timing of the project is intended to celebrate the next, and last for 100 years, transit of Venus in the
Southern Hemisphere. The voyage of the Endeavour
1768–1771, took place in the 'Age of Reason', a time of innovation and scientific discovery in Europe. The journey's primary purpose was largely politically motivated – it was to chart the transit of Venus over the
Southern Ocean, as an aid to British naval navigation, and reflected contemporary Anglo-French rivalry in establishing control of the oceans and discovering new colonies. The expedition was deemed so important that all other British naval vessels and captains were ordered to render Cook any necessary assistance. A second, secret set of orders, instructed the Endeavour to proceed south in search of the suspected but as yet undiscovered land mass known as Terra Australis Incognita, and to study and make collections of all natural materials, beasts, fish and minerals found. Although these
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instructions related only to the secondary aim of the voyage, it was because of them, and due to the passion and enthusiasm of the natural historians on board, that the whole expedition became so famous, and even today still fires the imagination. Visiting four continents and many isolated islands, the Endeavour voyage resulted in many new scientific discoveries, particularly in Australia and New Zealand. It also marked the first major encounter between indigenous people in these places with Europeans. Both the scientific discoveries made and the cultural encounters experienced, were to have a lasting impact on each other. This project will create a web portal dedicated to Joseph Banks & the Endeavour voyage. Central to this portal will be the botanical collections which will be re-united for the first time in almost 250 years, in a virtual space and made freely available. Much information is already available on
Banks or the voyage, but the actual collections themselves have always remained hidden away, in
Museums across the world. All the botanical specimens, and artwork from the voyage, will be digitised and be available for study. The portal will also act as a first point of reference for the voyage, including relevant maps, extracts from diaries and manuscript material, all directly linked to the specimens. Links to other sites which already cover other aspects of the voyage, will also be included. A second aspect of the project is the intention to place the collections in a wider context, and to explore and include as much indigenous perspective as possible, building on links which already exist. This part of the project will be exploratory, with the intention of building upon this aspect of the resource in the future. This presentation will explore some of the aims of the project in more detail, and also provide relevant background.
Progress to date will also be covered.
Reusable science
Whitbread, G 1 , Croft, J 1
1
Australian National Botanic Gardens and Australian
National Herbarium, Canberra, Australia
Authoritative lists of scientifically defensible names are the backbone of biodiversity science. The National
Species Lists project of the Atlas of Living Australia
(ALA-NSL) provides this key infrastructure of reusable nomenclatural and taxonomic content, and systems and services to enable interoperability within biodiversity informatics. The NSL project is:
• an integrated, open-data repository;
• owned and maintained by the taxonomic community;
• based on published or citable sources;
• based on web-based distributed data management solutions;
• built on biodiversity standards-based web services;
• using an architecture that accommodates both reusable open factual content and taxonomic overlays;
• supportive of content evolution through incremental change;
• able to ensuring persistence of cited extracts; and
• able to integrate with global initiatives.
The core of the NSL is provided by the Australian
Faunal Directory (AFD), maintained by the Australian
Biological Resources Study (ABRS), and the Australian
Plant Census (APC) based on the Australian Plant Name
Index (APNI) maintained by the Council of Heads of
Australian Herbaria (CHAH). These databases are the most complete on-line resources of information on the nomenclature and taxonomy of Australian plants and animals. They are of high quality, authoritative, actively maintained and widely accepted as the single point of truth for Australian nomenclature and taxonomy.
Complementary additional content comes from
Australian taxonomic compilations such as the
Interactive Catalogue of Australian Fungi , the Australian
Moss Catalogue , the Checklist of Lichens and the
Australian Marine Algae Index .
A key component of the NSL project has been the engagement of the Australian taxonomic community in compiling, extending and enhancing content to build the essential NSL infrastructure. The importance of this community engagement can not be underestimated; without it the NSL will not survive as a contemporary and on-going resource. Re-usability of the science content of the NSL reduces duplication of taxonomic effort, and increased efficiency in the shrinking cohort of practicing taxonomists. Existing services of the NSL include:
•
Persistent identifiers
•
LSID resolution
•
OAI-PMH and linked data objects for names, taxa, publications, classifications and people using TDWG standard forms for RDF, XML,
JSON and HTML
• name search incorporating fuzzy matching; and
• taxon name resolution services.
Taxon names are often used as convenient handles for taxa but are rarely anchored to cited concepts or hypotheses of the precise taxon in question; the connection is implied by context and prior knowledge and left as an exercise for the reader to associate data, information and knowledge. The NSL services address this problem by delivering reusable objects for scientific names and taxa that may be easily incorporated into digital works in flexible formats. Such objects substituted for simple names and concepts in online publications will provide for effective citation of the taxonomic authority wherever they are reused.
Advancing the International Plant Names Index
(IPNI)
1
Nicolson, N 1 , Paton, A
P 3 , Whitbread, G 2
1 , Croft, J 2 , Macklin, J 3 , Morris,
Royal Botanic Gardens, Kew, UK; 2
Herbarium, Canberra, Australia;
Herbaria, Cambridge MA, USA
3
Australian National
Harvard University
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The 'names and taxa' information space is often thought of as being composed of three layers: taxonomic concepts; Code-governed nomenclatural acts; name occurrences. In many circumstances the distinction of these layers is blurred, leading to confusion and inefficiencies in information management. To date, IPNI has been mainly concerned with the middle layer comprising ICBN governed nomenclatural acts, and is formed of three key components: curated data, information services to expose this data, and dedicated editorial staff to provide nomenclatural expertise. IPNI will be advanced from its current state to better connect to the layers above (taxonomic concepts) and below
(name occurrences). This will require the expansion of data holdings, improved linkages, and the development of information services and associated workflows. These will be offered to key actors including name authors, publishers, taxonomists and managers of biodiversity information.
New developments in the Integrated Taxonomic
Information System (ITIS)
1
Guala, G 1
United States Geological Survey, Core Science Systems,
USA
The mission of the Integrated Taxonomic Information
System (www.itis.gov) is to build and maintain a scientifically credible and publicly accessible database of taxonomic data and information on the world’s biota, placing priority on the taxa of North America. The presentation will focus on numerous major new updates including a comprehensive update for all North
American vascular plants as well as more geographically and taxonomically broad updates within the plants and other groups such as pollinators. There will be a demonstration of the New Online Taxonomic Workbench system designed to facilitate quicker updates and broader community involvement, and a discussion of major collaborations and capabilities.
Plant name services using Tropicos
Miller, C 1 , Freeland, C 1 , Magill, R 1 , Paige, J 1
1 Missouri Botanical Garden, St Louis, USA
The importance of organism names to global biodiversity informatics is becoming more apparent as development of the Global Names Architecture progresses. Resolving plant names and concepts has been particularly difficult on a global scale due to the lack of comprehensive online databases for plants. Catalogue of Life contains less than
50% of all angiosperm names. With over 1.2 million names tied to 119,000 references, 3.9 million specimens, and a data structure that enables multiple taxonomic opinions for synonymy and classification to be recorded,
Tropicos provides a valuable tool to botanists world-wide for plant name finding and resolution. Tropicos has been used as a source for plant names by uBIO and the Global
Names Index. The Biodiversity Heritage Library uses
Tropicos names for its taxon finding tools. Tropicos has also supplied name and concept data to the new Taxon
Name Resolution Service project at the iPLANT
Collaborative. The MBG Center for Biodiversity
Informatics has an ongoing effort to provide web services directly from Tropicos for name finding, taxon resolution, and name/taxon validation. This presentation will review the underpinnings of Tropicos and its plant name services. How Tropicos services are used to support biodiversity informatics, plant science, conservation, and sustainability projects worldwide will be discussed and plans for the future directions of
Tropicos support for name services will be described.
What's in a name? Not as much as you might think
(or like)
1
Stevens, P 1
Missouri Botanical Garden, St Louis, USA
Plant names have been called metadata, presumably signifying the value of the information accessible if names can be reconciled. However, much older botanical literature is bedeviled by hard-to- understand information that cannot be linked to specimens, and so any value is severely compromised when the circumscription
(geographic, taxonomic) of taxa is at issue. It can even be argued that there are few or no data, i.e. taxonomic information linked to specimens. Much taxonomic practice effectively precludes information from readily cumulating, the real taxonomic impediment; GenBank, for all its problems, has taken off in part because the information there is ultimately linked to specimens.
Recommendations include routinely putting specimen measurements in freely accessible databases, rethinking the practice and purpose of taxonomic monographs, aggressive use of nomenclatural instruments such as names in common use, promoting the use of a common plant ontology, and the institution of new starting dates for as many groups as possible. The use of older literature should be optional, not mandatory. Vivat
Linnaeus! Really.
Biodiversity Heritage Library: a global resource for
1 open access scientific literature
Freeland, C 1
Missouri Botanical Garden, St Louis, USA
The Biodiversity Heritage Library (BHL) is an international consortium of more than thirty of the world’s leading natural history museum and botanical garden libraries, working together to digitize the public domain literature in their collections. BHL also works with rights holders within learned societies and other organizations to obtain permission to digitize their journal backfiles and monographs, enabling the inclusion of contemporary content in BHL while still publishing all materials via open access means. The digital images and derivative access files of scanned books produced through BHL digitization activities are freely available for download and reuse by anyone with an internet connection. In this way BHL provides new digital access
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to content that has previously only been available in bound form on library shelves, and does so with minimal restrictions. In addition to publishing the digital files representing a book, BHL has built services and APIs on top of its corpus that allow content to be enhanced through manual data entry and automated analyses. BHL provides interfaces for authorized librarians and users to edit bibliographic metadata for scanned books, including the ability to represent titles in ways that they have been referenced by scholars and various indices since
Linnaeus’ publications in the 1750s and before. BHL has built and enhanced existing user interfaces for viewing the digitized books online, including services that enable third-party software developers to bundle BHL content within their own applications. BHL also employs the use of external services to enhance its registry, such as its incorporation of the TaxonFinder service from uBio that searches for and extracts all of the scientific names from the BHL corpus, providing users with an innovative names-based index to more than 250 years of scientific literature. BHL is leading a new initiative to mirror the digitized volumes in its collection at global scale and to bring in new content by partnering with large organizations and multi-national consortia that can provide content and infrastructure to BHL through their own mission-funded or grant-funded activities. Through these partnerships BHL is building an internationally replicated digital library of taxonomic works, serving students and scholars researching the world’s biodiversity as well as computer scientists and informaticians building new and novel algorithms for the semantic enhancement of scientific texts.
Link yourself or perish? Presentation of
PhytoKeys
,
1 the next generation journal in systematic botany
Penev, L 1 , Cress, WJ 2 , Knapp, S 3 , Li, D-Z 4
Pensoft Publishers, Sofia, Bulgaria;
Institution, Washington DC, USA;
Museum, London, UK; 4
3
, Renner, S
2
5
Smithsonian
Natural History
Kunming Institute of Botany,
5
Chinese Academy of Sciences, Kunming, China;
University of Munich (LMU), Germany
The presentation describes the focus, scope and the rationale of PhytoKeys (www.phytokeys.org), a newly established, peer-reviewed, open-access journal in plant systematics. PhytoKeys is launched to respond to four main challenges of our time: (1) Appearance of electronic publications as amendments or even alternatives to paper publications; (2) Open Access (OA) as a new publishing model; (3) Linkage of electronic registers, indices and aggregators that summarize information on biological species through taxonomic names or their persistent identifiers (Globally Unique
Identifiers or GUIDs; currently Life Science Identifiers or LSIDs ); (4) Web 2.0 technologies that permit the semantic markup of, and semantic enhancements to, published biological texts. The journal will pursue cutting-edge technologies in publication and dissemination of biodiversity information while strictly following the requirements of the current International
Code of Botanical Nomenclature (ICBN). The journal provides mandatory inclusion in the publication of IPNI records for new taxonomic names and other nomenclatural activities. All new species are supplied to the Encyclopedia of Life simultaneously with publication
332 of the article; all taxon treatments are deposited on Plazi
(www.plazi.org). The content of the article itself is marked-up at the level of taxonomic descriptions, taxon names, citations and references, identification keys, georeferenced localities, and other taxon information. All papers are open access and free to read, download, print, and distribute. PhytoKeys is more than a journal. It is a linked environment built upon its own content management software and an XML-based editorial workflow. Linking is provided at the internal level
(within an article, within the journal, or within the publishing platform of Pensoft) and to external resources
(GBIF, EOL, BHL, NCBI, Morphbank, IPNI, PLANTS,
Tropicos, The Gymnosperm Database, Index
Herbariorum, Wikipedia, Wikispecies, etc.) through a dynamic web profile of each taxon mentioned within a paper (www.ptp.pensoft.eu). Geo-referenced localities can be mapped within taxon treatments or for the entire paper. The journal can be followed on Twitter, Facebook,
Mendeley, and several other social networks. PhytoKeys provides workflow for data publication through GBIF's
Integrated Publishers Toolkit (IPT) and accepts manuscripts generated in XML from Scratchpads or authors' databases. PhytoKeys is published in four different formats: (1) high-resolution, full-color print version (2) PDF identical to the printed version; (3)
HTML to provide links to external resources and semantic enhancements to published texts for interactive reading; (4) XML version compatible to PubMedCentral archiving (based on TaxPub schema, www.sourceforge.net/projects/taxpub), thus providing a machine-readable copy to facilitate future data mining.
Neither restriction nor charges are imposed on the use of color illustrations.
Taxonomic Literature
(
TL-2
) as a major index to help guide selection and prioritization of botanical literature digitization
Dorr, LJ 1 , Everly, R 1
1 Smithsonian Institution, Washington DC, USA
Taxonomic Literature ( TL-2 ) is a multi-volume print publication that has become an indispensable and essential resource in supporting the research of systematic botanists (and zoologists) and an important reference tool in the science information field for librarians and information professionals. The work has been published over a span of 32 years by five principal authors. As a guide to the literature of systematic botany published between 1753 and 1940, the volumes include over 37, 000 titles published by 9,072 authors. Its coverage is international, providing bibliographic analysis to works in several languages such as German,
English, French, and Latin. Overall, TL-2 provides the most comprehensive biographical and bibliographical analysis for systematic botany literature prior to 1940. Dr
Laurence J. Dorr, Research Botanist & Associate Curator in the Dept of Botany, NMNH, Smithsonian Institution as well as the co-author of the two most recently published volumes, will give a historical perspective, describe how the publication is utilized by scientists and discuss its importance to the practicing taxonomist. Ms
Robin Everly, Branch Librarian, Botany–Horticulture
Library, Smithsonian Institution Libraries (SIL), will present the current project to improve accessibility and

utility of TL-2 . The print publication is being converted and enhanced into a searchable database with multiple access points by the Digital Services Division, SIL. Once created, the TL-2 database will be available for open access through the Biodiversity Heritage Library website.
A schema for description and exchange of taxonomic publication's content
Agosti, D 1 , Catapano, T 2
1 Plazi; 2 Plazi /Columbia University, New York, USA
The increasing data deluge makes it increasingly difficult to find the piece of information that is wanted. Even in our limited world of biodiversity literature millions of pages of printed records are made accessible parallel with a rapidly growing amount of Web-content, from digital scientific publications to dedicated Web sites to a multitude of popular online articles covering our subjects. Whilst finding pages with content we are after, such as a particular taxon, has been facilitated by services like Google, the results are so overwhelming that the return becomes counterproductive, not least because finding names is simple, and the majority of the content is not what is expected. In other words, the search is not specific enough because it doesn’t cover the terms’ expected context. One way to get around this impediment is to add some little information in the text that explains what the text is about. Similar to the ubiquitous and to most Internet users familiar Hyper Text Markup
Language (html) that formats a text so it appears in a particular way on the screen, the Extended Mark-up
Language (XML) allows to define blocks of text to reflect what content is within pieces of text, such as a description of a taxon, a synonymy or a character. This allows defining domain specific elements on the one hand, and on the other the machine to find particular tags and its content, and in a more advanced way to infer relationships, such as that everything within a treatment delimited block of text is in relation to the nominate taxon of this treatment. It also offers the potential to markup requested elements, such as all those covering the information needed to create a nomenclatorially valid taxonomic name. Finally it allows creating services that allow machines to exchange particular elements of texts we are interested in. In this lecture we will provide a overview of the principles of a taxonomy specific XML schema, TaxonX for the mark-up of legacy literature and a derivative, Taxpub for prospective publications. Both of them are being used to mark up legacy publications and to publish taxonomic work respectively. The examples are drawn primarily from work done at Plazi in collaboration with the Global Biodiversity Information
Facility and support of the US National Science
Foundation and Deutsche Forschungsgemeinschaft, and the US National Institutes of Health and Pensoft.
Names-based retrieval of library and internet resources in the Virtual Library of Biology (vifabio): issues in using taxonomic names in library catalogues and metasearch tools
Kasperek, G 1 , Rexhepi, J 1 , Daehne, J 1
1 University Library Johann Christian Senckenberg,
Germany
The portal 'Virtual Library of Biology' (vifabio, see http://www.vifabio.de/?lang=en) offers centralised access to literature and information in libraries and on the
Internet, for biology in general and for taxonomy in particular. The Virtual Catalogue provides bibliographic records from many databases which are searched in parallel (metasearch), including catalogues of numerous specialised libraries, bibliographic data of the
Biodiversity Heritage Library and other digitisation projects, several article databases as well as the vifabio
Internet Guide. Because the taxonomic relevance of a certain publication does not depend on online availability, vifabio places an equal emphasis on digital and printed-only materials. With bibliographic records, users of vifabio will find various options for access whenever possible. Most printed-only materials are made available worldwide through document delivery services. vifabio is operated by the University Library 'Johann
Christian Senckenberg' in Frankfurt am Main (Germany), in conjunction with other libraries and biological organisations. vifabio is on the Web since 2007, and its search portal is available at no cost for users worldwide
(except for document delivery services). The presentation will highlight issues in using taxonomic names in library catalogues and bibliographic databases. Advantages and shortcomings of metasearch will be discussed, such as technical issues of connecting several library catalogues and databases with heterogenous data structures. Some consequences for further developing vifabio's search tools will be debated. The presentation will close with some considerations on implementing 'taxonomically intelligent' names-based search tools in library environments. URL:http://www.vifabio.de/?lang=en
1
Fine-grained semantic markup of descriptive data
Cui, H 1
University of Arizona, USA
Taxonomic literature contains rich descriptive data about various organisms. Such literature ranges from reference works such as Flora of North America and Flora of
Australia to scientific articles published in journals such as ZooTaxa . The descriptive data are valuable to systematic biology research, phylogenetic research, ecology and conservation, and climate change studies.
However, because the vast majority of the descriptive data are written in a semi-natural language form and published in a human readable format such as print text, web pages, and PDF, they are not easily accessible by computers, making locating and mining descriptive data challenging. In the past five years, we have been developing supervised and unsupervised machine learning methods to convert semi-natural language descriptions to structured digital formats such as XML
(eXensible Markup Language), SDD (Structured
Descriptive Data), and RDF (Resource Description
Framework). Our experience shows that unsupervised methods are less expensive and equally effective, compared to supervised methods, as the latter requires hundreds or thousands of training examples. A set of unsupervised machine algorithms have been created and some of them have been tested on document collections such as Flora of North America , Treatises on
Invertebrate Paleontology and OCRed documents from the Biodiversity Heritage Library. The algorithms were
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not designed to process a specific document collection, but are for the markup of all descriptive data (plants or animals) in semi-natural language form. In this abstract, we briefly introduce three algorithms. The unsupervised paragraph extraction algorithm learns to identify paragraphs that contain morphological descriptions from their parent documents. This algorithm is useful as descriptive data is often presented side by side in a document with other types of information such as background information, research methods, and discussion paragraphs. The unsupervised clause markup algorithm bootstraps organ names and descriptive terms from morphological descriptions. This algorithm not only marks up a description sentence by sentence, it also learns organ/structure names and character states. Since comprehensive dictionaries covering all organs/structures and/or all character states do not exist, this algorithm is useful to propose new terms to be admitted to an open accessible dictionary or ontology such as PATO
(Phenotypic Quality Ontology). Ontological control of structure names and character state values are crucially important to fine-grained markup of descriptive data, especially to the markup at characters and character states level. The algorithm we developed for character level of markup employs all above mentioned algorithms and relies on the unsupervised clause markup algorithm to provide correct Part of Speech tags to natural language parsers such as Stanford Parser and to segment a sentence to separate semantic units. The character level markup annotates organ/structure names, characters, character states, and relations between organs/structures.
For example, a sentence 'Individual heads sessile or each borne on a peduncle ;' is marked up in XML format as
<statement id='s1.txt-8'> <structure id='o22' name='head'> <character name='architecture' value='sessile' /> <character name='architecture' value='each borne on a peduncle' /> </structure>
<structure id='o23' name='peduncle' /> <relation id='r8' name='borne on' from='o22' to='o23' /> </statement> If time permits, we will demonstrate the markup system live at the conference.
Is there a dynamic future ahead for Floras?
Wilton, A 1 , Breitwieser, I 1
1 Landcare Research, Lincoln, New Zealand
Floras are an authoritative summation of taxonomic knowledge for a given region at a given time. They provide an important portal for users both inside and outside the taxonomic community to access taxonomic information. In a typical scenario, ongoing taxonomic work produces revisionary papers and curated census and specimen databases. At a given time, data from these and other sources are crystallized into a Flora, usually published in hardcopy. Revisionary and herbarium curatorial work continues, resulting in a growing mismatch between the true state of knowledge and that represented in the Flora. At infrequent intervals, new
Floras or new editions of existing Floras are produced, and the process continues. Because of the considerable
334 time and resources required to compile and produce a
Flora, being out-of-date is the norm rather than the exception – sometimes even before a Flora is printed. It is also typical for a Flora to be produced as a single product – traditionally as a printed document – that is fully accessible to only limited audiences and costly to transform for other audiences. This mismatch between actual knowledge and the Flora, and the way it is delivered, is a significant problem to users, who increasingly require rapid and timely access to up-to-date information in a format that meets their requirements.
The Flora of New Zealand project is overcoming these problems. It has the goal to deliver the next generation of
New Zealand Floras as a dynamic resource: a resource that is both up-to-date and engages a wide range of users.
This involves changing the Flora creation process to develop an information system that allows integration of data from a range of sources, and to extend the concept of a Flora to include its delivery via a variety of tailored products – the first being a version delivered via the internet. Central to achieving this is the development of processes that allow the capture and analysis of granular, highly linked data to ensure that data used to create the
Flora are up-to-date. This creates a number of technical challenges such as efficient processes to capture data linked to specimens, and the development of processes to analyse these data, as well as social and legal challenges such as changing working practices by scientists and the recognition of authorship and copyright.
Floras in the 21st Century: the
Flora of North
America
saga
2
1
Macklin, J 1 , Cui, H 2 , Morris, R 3 , Morris, P 1
Harvard University Herbaria, Cambridge, MA, USA;
University of Arizona, Tucson, USA; 3 University of
Massachusetts-Boston, Boston, USA
The Flora of North America (FNA) has now published more than half of its intended 30 printed volumes. The reality of a project that has spanned more than two decades is that many treatments are well out of date based on new science before the project is even finished!
Floras are dynamic by nature, both due to changes in our understanding of the relationship and delineation of the groups treated, and the appearance and disappearance of species in the treated area over time. Thus, flora should be built and presented in a dynamic environment as well.
The first challenge is converting the printed word into discrete digital objects that can be reassembled in various useful ways by software. An ongoing collaboration to build intelligent text parsing tools for legacy literature has allowed us to achieve fine-grained parsing of treatments to the character and attribute level. These characters can then be managed in a semantic controlled vocabulary (glossary) allowing authors to build new treatments that are automatically comparable and parallel to other treatments. This semantic relationship allows for more efficient creation of treatments, more complex and powerful searching of their content, and production of interactive keys on-the-fly. We are developing a demonstration web portal which incorporates these parsed FNA treatments with knowledge from many relevant aggregated data sets of occurrence records and metadata such as the Global Biodiversity Information
Facility (GBIF), United States Dept of Agriculture–

PLANTS, the U.S. National Invasive Species
Information Center (NISIC), NatureServe, the U.S.
National Biological Information Infrastructure (NBII), images in Morphbank, research-specific databases, and millions of digitized records from specimens housed in herbaria. The knowledge from all of these sources will be managed by a Filtered Push network that provides annotation services and an ability to syndicate or push updates and revisions both to and from interested parties.
At some point, this dynamic presence may be used to synthesize a new stable treatment version through the use of a work flow management tool with the interaction of authors, reviewers and editors, as was done when producing the books. Using these tools will allow authors to produce a treatment much more quickly, and will help manage the process towards attributable e-publication of treatments. Users of the portal can choose whether to interact with the dynamic, but potentially nonauthoritative, content, or with the most recent stable treatment. They can then access this content, annotate it, and produce information as a synthesized digital Flora of a given group or location, including illustrations and photographs, maps, and interactive keys, or use a printon-demand service. This new portal and tools could fundamentally shift the way that Floras are created through a constant cycle of asynchronously updating as demand or interest dictates, making this resource invaluable to professional botanists, amateurs and the general public, alike.
The
Flore d’Afrique Centrale
in the context of the
'European e-Floras Initiative'
Janssen, T 1 , Sierra Daza, S
Smets, E 2 , Robbrecht, E 1
2 , Dessein, S 1 , Roos, M 2 ,
1 National Botanical Garden of Belgium, Domein van
Bouchout, Belgium; 2 Netherlands Centre for Biodiversity
Naturalis (section National Herbarium of the
Netherlands), Leiden University, The Netherlands
Timely elaboration and dissemination of information on plant diversity is crucial to identify, understand, use and conserve plant resources. A growing demand for electronic and online floristic data has led to a proliferation of e-Flora and e-taxonomy projects. Current e-Floras facilitate access to existing, but not the production of new content and there is limited interoperability. Supported by a large consortium of
European botanical institutions, the „European e-Floras
Initiative' aims at improved communication and collaboration in the taxonomic research community to adapt work processes, maximize interoperability and establish best practices for the use of recently developed
IT tools and data models to compile e-Floras. The Flore d’Afrique centrale ( Flora of Central Africa ), published by the National Botanic Garden of Belgium, is the only comprehensive source of botanical information on
D.R.Congo, Rwanda and Burundi, an area that includes the world’s second largest rainforest. Between 1963 and
2005, 87 fascicles for flowering plant and fern families were published in print covering about 60% of the known species of vascular plants from the region. The project currently relies on an increasingly scarce and distributed community of taxonomic experts. We here comment on the progress made in the framework of the ‘European e-
Floras Initiative' in establishing an electronic edition of the Flore d’Afrique centrale based on the EDIT Platform for Cybertaxonomy, complementing the printed edition, and offering multiple services for data providers and data users. The data provider services include infrastructures for remote collaboration on new treatments and data curation as well as direct print and online publication from the Flora database. The data user services include advanced key and visualisation technologies as well as commenting and discussion tools for non-expert data annotation and community building. With this approach we aim at (i) providing an efficient, interoperable, economically attractive, and trustworthy medium to distribute plant knowledge on Central Africa and (ii) achieving completion of the Flore d’Afrique centrale in a timeframe reflecting the high conservation concern of the
Central African plant diversity.
Online interactive pathway (directed) keys: an effective tool for serving today's students while
1 building resources for tomorrow's students
Barkworth, M 1
Intermountain Herbarium, Utah, USA
The primary function of Floras is to provide information about the taxa in a region but they are also important educational resources. It is through working with a regional Flora that students and others become familiar with taxonomic groups, or at least those in their region.
Unfortunately, many existing regional Floras reflect badly outdated taxonomic concepts. Consequently, today’s students, who are tomorrow’s botanists, are being told about new taxonomic concepts but forced to work with old ones, a dichotomy that is frustrating to all involved and contributes. Creation of illustrated, webaccessible multi-access interactive keys that reflect current concepts are one solution to this problem, but producing them takes considerable time. An alternative approach, one being used in the Intermountain Region of western North America, is to provide illustrated, webaccessible pathwayS. Such keys can be constructed relatively rapidly for regions with existing treatments and, as with multi-access keys, the taxon names can be linked to pages that cite existing resources and/or connect to web pages providing additional information, including information generated from constantly updated databases. Two other advantages of directed-choice keys are that (1) through repeated used of the same pathway, users become familiar with the distinguishing characteristics of the taxa, particularly as these apply to the region concerned; (2) the keys can be made available in print format, a format that many active field botanists prefer; and (3) they encourage users to become familiar with structure of the organisms concerned. Their disadvantages are that (1) creating pathway keys does not result in creation of a descriptive database; (2) incorporation of new taxa and taxonomic changes is not quite as simple as it is with free-choice keys (although much easier than developing a whole new printed Flora);
(3) they cannot be used for identification of fragmentary material; and 4) they cannot be used by people with minimal understanding of plant structure. Nevertheless, their ease of development in areas having good floristic resources as a starting point makes them a practical mechanism for accelerating the movement of systematic
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research results to users, including the generation of botanists now in our classrooms.
The eFlora of Anywhere and the eFlora of
Everywhere – Floras in a highly connected world
Thiele, K 1
1 Western Australian Herbarium, Australia
A Flora, as traditionally understood, is a summary of systematic knowledge for a given area. Thus, we have the Flora Europaea , the Flora of Australia , the Flora of
China, etc. When Floras had no choice but to be on paper, this was the only sensible strategy. Early and current generation eFloras have generally adopted this model, sometimes via digital representations of existing paper Floras, sometimes starting from scratch. I argue that this model has a limited shelf-life. In the highly connected world of the 21st Century, we need a new model in which an eFlora of a given region is created by aggregating existing material from a wider variety of specialised data sources, including other eFloras. One way of looking at this is that we need to build the eFlora of Anywhere – a local view of the eFlora of Everywhere
– rather than a whole bunch of eFloras of Somewhere.
Curiously, though, we can also build the eFlora of
Anywhere/Everywhere by first building a whole bunch of eFloras of Somewhere, so long as we go about it in the right way. This talk will discuss the elements of modern, connected eFloras and provide a model for the eFlora of
Everywhere.
Limits and potentiality of geometric morphometrics in plant systematics
Passalacqua, NG 1
1 University of Calabria, Italy
Geometric morphometrics methods started to be developed in the late 1980s. During the 1990s, it showed a remarkable qualitative and quantitative development, thanks to the increased understanding of the theoretical basis that underlie the methodologies, the development of protocols of analysis, and the consequent increase in published works that referred to these methods, especially in Zoology and Anthropology. During this time progress in botanical applications has been marginal indicating a reluctance to accept new methods for morphometric analysis. While we cannot exclude a certain cultural inertia by botanists, it is undoubtedly true that the study material of botanists, i.e. plants, is usually less suitable for the application of geometric morphometric techniques. However, in recent years, experiments carried out by botanists gave a glimpse of how these techniques can offer considerable support to systematic studies in Botany. This presentation will first analyze the main difficulties related to the use of geometric morphometric techniques with plants, with particular reference to the method of landmarks; limits we need to be aware of for a correct application of these
336 methodologies and interpretation of results. The presentation will then move on to the observation of situations where geometric morphometric techniques showed significant potential for exploration and/or interpretation in plant Systematics.
The use of geometric morphometrics in evolutionary
1 studies
Rohlf, FJ 1
Stony Brook University, New York, USA
A discussion of some of the statistical properties of shape data provided by geometric morphometric studies as well as some of its limitations for use in taxonomic and evolutionary studies will be presented. While in many ways there is nothing special about the multivariate analysis of such multivariate data, such data does have to be treated properly. An important property of such data is that it makes possible new ways of visualizing the results of multivariate analyses.
Integrating morphometric and phylogenetic analyses: from phenetic systematics to phylogenetic morphometrics
De Luna, E 1 , Chew, T 1
1 Biodiversidad y Sistemática, INECOL, AC, Mexico
The role of morphometric analyses in plant systematics has stirred mixed reactions. Applications such as discriminant analyses for taxon identification, or fitting morphometric data to a phylogeny to estimate ancestral shapes and changes along nodes on the tree do not seem controversial. What remains unsettled is if morphometric analyses should help and how in decisions about characters, definition of character states, and for taxonomic grouping. Morphometric studies can be a strong basis for phylogenetic reconstructions. A sound integration of morphometric data in phylogenetic analyses is by the use of multivariate methods not for the circumscription of taxa, as in a phenetic approach, but for questions about character homology and monophyly. A double path is now open for the integration of morphometrics into phylogenetic analyses depending on what is considered as 'cladistic character' and how to analyze continuous patterns of variation in morphological variables. The first question is what counts as character data: whole shape, separate modules, partial warps, relative warps? The second question is whether variation must be partitioned and coded as discrete states or is analyzed as continuous data. We argue for a need to move from phenetic systematics to phylogenetic morphometrics. We can now integrate very nicely morphometric data into into phylogenetic reconstructions of monophyletic groups by using continuous characters and landmark data without coding or transformation.
Empirical tests show that the use of relative warp scores, either means or ranges as continuous characters, are to be preferred for phylogenetic analyses based on criteria of concordance with non morphometric trees, levels of homoplasy, and clade resolution/support.

Reconstructing ancestral floral shape using geometric morphometrics: a test of the pollinator shift hypothesis
Van Der Niet, T
MS 3 , Johnson, SD 1
1,2 , Zollikofer, CPE
, Linder, P 2
3 , Ponce de Leon,
1 School of Biological and Conservation Sciences,
University of KwaZulu-Natal, Pietermaritzburg, South
Africa; 2 Institute of Systematic Botany, University of
Zurich, Switzerland; 3 Anthropological Institute and
Museum, University of Zurich, Switzerland
Variation in floral shape has been used for centuries as a diagnostic tool in plant systematics. The absence of adequate methods to quantify floral shape, however, has inhibited research into the underlying evolutionary processes that generate this variation. The recent developments of a rigorous statistical framework for comparative analyses of shape, and the ability to produce
3D reconstructions of flowers with complex shapes, have opened the door for addressing a wide variety of evolutionary questions involving floral shape. We explored the role of pollinators in the evolution of floral shape variation in the orchid genus Satyrium , the species of which have remarkably complex flowers, which are characterized by specialized interactions with a wide variety of insect and bird pollinators. We produced 3D reconstructions of flowers using microCT scanning and obtained 40 homologous landmarks and semi-landmarks.
A principal components analysis of a data set in which size was removed by translation, rotation and scaling, showed that species with the same pollinator have highly similar shapes. To remove potentially confounding phylogenetic effects, we first reconstructed ancestral shapes onto an ultrametric tree by applying squared parsimony to the procrust coordinates. We then calculated the rate of evolution of shape by calculating the Euclidean distance between reconstructed shapes at the base and tip of individual branches, and by dividing this by the branch length. Finally, we reconstructed pollinator shifts onto the branches of the phylogenetic tree. We found that the rate of morphological evolution is significantly elevated in branches where there is an unambiguous optimisation of pollinator shifts, compared to branches where there is unambiguously no pollinator shift, thus confirming the long-standing notion that pollinators have played an important role in the diversification of angiosperm floral shape. Although our analysis is intuitively appealing, it assumes that, in the absence of selection, evolution of quantitative traits occurs according to a model of Brownian motion. Our initial finding of strong similarity among species with the same pollinator, however, suggested that floral shape is a highly adaptive trait. This begs the question whether it is valid to use a model of essentially neutral evolution for the reconstruction of ancestral floral shape. Closer examination of ancestral shapes showed that in cases of a pollinator shift between sister taxa, the shape of the immediate ancestor was intermediate as expected from
Brownian motion. These intermediate shapes sometimes fell outside any of the well-defined, pollinator-specific clusters in the principal components analysis. This means either that these 'hybrid' shapes may be unfit, or that they represent adaptations to unknown pollinators. Either of these possibilities poses a challenge to accurate evolutionary analyses. The development of alternative models of evolution that allow for incorporation of adaptive peaks, such as Ornstein-Uhlenbeck models for multivariate traits, would provide a solution to this problem. Nevertheless, we believe that our novel approach of analyzing geometric morphometric data in a phylogenetic and ecological context holds great promise and represents a first step towards quantitative approaches to addressing long-standing evolutionary hypotheses.
Lessons from the Botanical Capacity Assessment
Project (BCAP) – building links between stakeholders
Kramer, A 1, 2
2
1 Botanic Gardens Conservation International US;
Chicago Botanic Garden, USA
The botanical community plays a mission-critical role in researching, conserving, and sustainably managing the world’s plant diversity and resources. Botanical expertise is required to address current and future issues, including climate change mitigation, land management and habitat restoration, maintenance of ecosystem services, control of invasive species, and conservation of rare species. Yet despite the fundamental role botanical capacity plays in tackling these issues, a recently published report in the
United States reveals severe shortages of botanists at government agencies, a wave of upcoming retirements, and an alarming decline in botanical degree programs and course offerings at the nation’s colleges and universities. The result of a year-long project which surveyed nearly 1,600 members of the United States botanical community, this report (entitled ‘Assessing botanical capacity to address grand challenges’) details the importance of building links between stakeholders throughout the botanical community. It describes how private sector organizations are helping to fill gaps in capacity being created by declines in the academic and government sectors, and urges action across all sectors to work more strategically to more effectively pool resources and ensure program sustainability and conservation success into the future. While this project was carried out in the United States, its results are internationally relevant and applicable. From Australia to the United Kingdom, similar but often unquantified reports of declines in botanical capacity have been circulating for decades. These declines are occurring as the need for botanical capacity rises ever-higher, particularly in developing countries where plant diversity and the need for identification, research, and conservation work is greatest. Report results illustrate the importance of working across public and private sectors to ensure botanical capacity is valued, understood, and sufficiently supported.
1
Does plant identification service shows any trend?
Sumadijaya, A 1
Herbarium Bogoriense, Indonesian Institute of Sciences,
Cibinong Science Center, Indonesia
337

Herbarium Bogoriense (BO), as an integrated part of
Botanical Division, Research Center for Biology,
Indonesian Institute of Sciences, daily receives specimens for identification service. As the Scientific
Authority of Biodiversity in Indonesia, the identification service provides each user with accurate species names.
Specimens are derived from many regions in Indonesia, variable parties, and many purposes. Five years data, from 2005 to 2009, has been extracted from thousands of specimens and users. We hope to see a variety of trends based on time (year, month), purpose (research, business or other), user (institution, private, or other), user location, occupation (students, staf or other) and other observable. Moreover, we hope the data shows trends on the dominant families, genera and species levels. Output from the analyses are expected to build the grand scheme of botanical research in Herbarium Bogoriense.
Strengthening the foundations of plant conservation in Brazil: perspectives of a new decade
D`Ávila De Moraes, M 1
1 Brazilian National Centre for Plant Conservation,
Brazil
In Brazil, Botany started to take place with the arrival of foreign scientists, especially naturalists, after the transference of the Portuguese Court, in 1808. Martius,
Saint Hillaire, Pohl, Gardner, Riedel and Glazious, among others contributed with collections and amazingly detailed journals about the flora. Brazil had an eminent colonial structure, where the general instruction level of its people was low, and just a few privileged ones could travel abroad to receive academic formation.
Consequently, it took a while before a National School of
Botany could be institutionalized. In 1822, Brazil became politically independent and, in order to consolidate the territory of the New Republic, the Government started organizing missions to occupy the inland and secure the borders. Meanwhile, naturalists were sent along with workers and the army. Despite few plant conservation initiatives, environmental concern was triggered after
1992. Rio de Janeiro hosted Rio Earth Summit, a meeting to discuss measures to stop environmental destruction.
During the event, the Convention on Biological Diversity was signed, which represented a milestone and generated a new demand regarding plant capacities. Besides describing the Brazilian plant diversity, the academy received the responsibility to define strategies and priorities for plant conservation, shifting from an exclusive botanic approach to a conservationist perspective. Nevertheless, the transition generated a gap fulfilled by the third sector. Since then, several environmental NGOs have been established in Brazil and started working specifically with government incapacities in the agenda. Important results were achieved such as international fund-raising, social engagement, networking and capacity building.
Nevertheless, the advances were never fully incorporated by the government, which still depends on the third sector to achieve goals related to environmental policies.
Despite achievements in the past decade, a backset has been led by the present government. Most flora conservation initiatives were implemented following international agreements in plant conservation. But the numbers are hiding a catastrophic scenario of
338 biodiversity loss, mostly attributed to incapacities of the federal agencies in dealing with the pressure for economical development and natural resources exploitation, besides conflicting policies and the absence of proper communication with the third sector. Farther on, a shortage of botanists trained to work with conservation biology is being foreseen by governments of several countries. However, the permanent increasing number of postgraduate students of botany constitutes a different scenario nationally. And despite new openings at federal agencies, NGOs have been offering better work opportunities, pushing recently graduated professionals and senior researchers to the third sector. From 2002 to
2005 the number of environmental NGO's has grown c.
70%, more than three times the national mean of growth of 22,6% for the whole sector. This trend is weakening government capacities and building dependency on the third sector. To avoid future shortages in human resources we should invest in institutional infrastructure creating opportunities under the Environmental Ministry and its agencies. In the new scenario Botanic Gardens may play a key role in promoting linkage among government, NGOs and the private sector, strengthening the foundations of plant conservation in Brazil.
Learning about plants in first year college/university
1
– a mix of the real and virtual worlds
Burrows, G 1
Charles Sturt University, Wagga Wagga, Australia
Botanical studies in first year college or university involve a diverse body of students and a wide range of material to be covered. Some students come with a love of plants and prior biological study, while others are required to take botany as part of a set degree structure and might have a limited science background. Most topics are covered in a single lecture and single practical session, with limited opportunities for revision before an end of session practical and/or theory exam. Given that a topic, e.g. flower structure, may be covered in a single week, at certain times of the year there may be limited material available to show a reasonable range of variation in important features, e.g. ovary superior/inferior, flowers actinomorphic/zygomorphic. In an attempt to overcome some of these limitations I have developed a series of web-based applications that cover some fundamental aspects of leaf and flower morphology (see http://www.csu.edu.au/herbarium ):
• Leaf morphology (alternate/opposite/whorled; simple/compound)
• Gynoecium guide (superior/inferior ovary; number and fusion of carpels)
• Floral symmetry (actinomorphic/zygomorphic)
• Virtual floral formula (integrating several of the other applications to construct floral formulae)
• Plant family recognition (recognising plant families ‘on sight’)
• Supermarket botany (using everyday plant food items to learn about stems/roots/leaves and the sequence of flowers/fruits/seeds),
All the applications feature a tutorial and tests based on high quality images of living samples. The tutorials feature one set of images, while the tests use different sets of images so that students apply their knowledge to a different data set. The tests are interactive, giving

feedback as to why an answer is correct/incorrect. At
Charles Sturt University about half of our first year botany students study full time with weekly practical sessions (internal students) and half are distance education students who receive their practical sessions in an intense four day residential school. For both cohorts these interactive activities address many of the limitations described above:
• Students can prepare for practical sessions. This is particularly useful for the distance education students as they make the most efficient use of their limited time on campus.
• Students can revise the material after practical sessions.
This is particularly useful for the full time students where it may be several weeks between a practical activity and the end of session exams.
• Students are able to observe and assess a wider range of material than can be studied in a practical period or than might be available at most times of the year.
All these applications are designed to supplement and support the hands-on, living plant material, practical sessions. Where ever possible we integrate the lectures, practicals and online material. Feedback indicates that students appreciate: the ability to prepare for and revise after practical sessions; the ability to study at their own pace and time; the feedback given in the tests; and seeing progressive improvements in their skills.
Botanical literacy and botanical capacity: linking online scientist mentors to promote thinking about plants
Hemingway, C 1 , Sundberg, M 2
1 Botanical Society of America; 2 Emporia State
University, USA
Science and society stand to reap multiple benefits from efforts to improve student understanding of and appreciation for plants. To prime the pipeline pump needed to rebuild botanical capacity and fill gaps across diverse workforce sectors, botanical education across grade levels must be strengthened. The state of botanical literacy is part of a systemic science literacy crisis in the
United States. Some underlying causes – low interest in plants, little exposure to plants in school, limited coverage of plant content in education standards – are unique to botany and its recent history in US schools.
Regardless of discipline, however, inquiry-based learning is strongly supported as a means to enhance student understanding. Today’s teachers also seek to engage their students in collaboration, communication, and innovation, the 21st Century learning skills for success in the global environment. Here we present key findings on the PlantingScience online learning community, which leverages cyberlearning to make science experts broadly accessible to students and teachers. Scientists volunteer as online mentors to create new opportunities for students and teachers to learn how science works and how scientific research is conducted on plants. Students experience science, and knowledge building within a scientific domain, as a community endeavor by conducting plant investigations in their classrooms and communicating online about their research with their peers and mentors. Teachers have options to co-develop and field-test inquiry modules in scientist-teacher teams and experience inquiry immersion with scientists in a summer workshop. Initiated in 2005 by the Botanical
Society of America, fourteen scientific organizations are now partners. Over 7,500 students and their teachers in
31 US states have thus far collaborated online with over
400 plant scientists (global connectivity allows mentors to participate from Canada, U.K., Turkey, India, and elsewhere). High schools account for over 60% of the classes, but the model has been broadly applied from middle school to college students, after-school clubs and international groups. Assessing effectiveness involves varied data sources (e.g., student pre- and post-tests, teacher and mentor surveys, online artifacts), with research components conducted by Texas A&M
University and BSCS collaborators. We have found that middle school students appear to be slightly more engaged in discourse with their mentors than high school students. Students post most frequently about research questions and predictions, while mentors post more about predictions and experimental design. All students tend to struggle with presenting and making sense of their data.
Teachers feel that mentors are more helpful to the students in both the early and final inquiry phases than the mentors perceive. Preliminary results showed statistical increases in student attitudes about enjoying studying plants and finding plant biology an easy subject.
Numerous students report improved understanding of photosynthesis, alternation of generation, and other conceptually difficult topics. While lessons learned on the impact of this scientist-student-teacher partnership are ongoing, we are confident that online scientist mentors can serve as a critical linchpin connecting the plant education and research communities and effectively supporting student thinking about plants.
Charles Darwin (1809–1882) and his studies on orchid flowers
1
Bernhardt, P 1 , Edens-Meier, R
Saint Louis University, USA
1
On the Various Contrivances by Which Orchids Are
Fertilised by Insects (1862) celebrates its 150th anniversary in 2012. We review and compare the research and observations Darwin and his colleagues made on the functions of floral organs in temperate and tropical species within the 19th century framework of botanical studies and horticultural fads. This line of inquiry explains why Darwin used orchid flowers as model systems to show descent through modification after publication of On the Origin of Species (1859).
Additions to orchid diversity and reinterpretations of floral architecture in the second edition of this book
(1877) occurred because Darwin expanded his correspondence and received publications from new colleagues working in latitudes with far richer orchid floras. Considering the impact both editions made on academics, naturalists, pollination biologists and even science fiction writers living on five continents, over 150 years, it seems appropriate to abandon previous criticisms that Darwin wrote an obscure book grounded in Teleology.
339

Pollination biology of
Cypripedium
and
Paphiopedilum
Luo, Y-B 1
1 State Key Laboratory of Systematic and Evolutionary
Botany, Institute of Botany, CAS, China
Up to now, 24 of 156 species of three genera in subfamily Cypripedioideae have been studied on pollination biology. Four primary conclusions could be made from those studies: (1) The pollination system of two major genera in this subfamily, Cypripedium and
Paphiopedilum , is highly diverse. Both the bee pollination system including bumblebees, sweet bees, and Athophora bees, and the fly pollination system including syrphid fly, fruit fly, and Calliphora fly, occur repeatedly in Cypripedium and Paphiopedilum . (2)
Cypripedium and Paphiopedilum have different deceive strategy to attract the pollinator. The former mainly rely on the complex compounds of floral fragrance while the latter depend mainly on the floral coloration and ‘curious furry warts’. (3) The exclusive floral architecture of both genera appears to function as the primary mechanical feature to limit the visitors acting as effective pollinators.
And, thus, a specialized pollination system is formed throughout Cypripedium and Paphiopedilum , probably within the subfamily Cypripedioideae. (4) Both
Cypripedium and Paphiopedilum have a trend of transition from bee pollination system to fly pollination system. This transition could be carried out through either the limitation of the pollination route size in the flowers or the limitation of the attraction of the floral colour or/and floral fragrance to the pollinators. (5) The floral morphological features similarity between the species of Cypripedium and subgenus Brachypetalum of
Paphiopedilum , such as P. micranthum , P. malipoense , are results of adapted to bumblebee or sweet bee pollination. This similarity could not be considered having phylogenetic implications.
Osmophores in the labellum of three
Ophrys species: structure and role in pollination of sexually deceptive orchids
Francisco, A 1 , Ascensão, L 1
1 Universidade de Lisboa, Faculdade de Ciências, Centro de Biotecnologia Vegetal - IBB, Lisboa, Portugal
Pollination in the Mediterranean orchid genus Ophrys is a rare event that had already intrigued Charles Darwin in the nineteenth century. Since then, many efforts have been made for understanding the highly specialized process of pollination by sexual deceit that has evolved in the genus. The rewardless Ophrys flowers attract only male bees or wasps as pollinators by means of a combination of olfactory, visual, and tactile signals provided by the distinctive labellum, which mimics the female and its specific sex pheromone. Through pseudocopulatory attempts with the labellum, sexually excited males remove pollinaria from the flower and could enable cross-pollination as visiting another Ophrys flower. Odoriferous compounds emitted by the labellum are considered the key elements presiding at the success of sexual mimicry. Chemoecological research has been conducted on Ophrys floral scents, although the precise site of biosynthesis of the pseudopheromones remains
340 unknown for most species. Moreover, little attention has been addressed to the role of visual and tactile stimuli in the attraction of pollinators. Here we compare in detail the labellum micromorphology in three closely related
Ophrys species ( Ophrys tenthredinifera , O. bombyliflora ,
O. speculum ) and provide anatomical and histochemical evidence for the presence of an osmophore in the labellum of all taxa studied. Scanning electron microscopical observations revealed a great diversity of epidermal cell types in the adaxial surface of the labellum in all species, forming a species-specific patterning. Most significantly, the three labella differ in the apical region where a central notch included in a broad glabrous border occurs in O. speculum , a glabrous recurved appendix is evident in O. tenthredinifera and a unique deflexed fleshy appendix is found in O. bombyliflora . Typically, dome-shaped papillae compose the margin of the labellum and a submarginal band of long contorted trichomes is present in all species. Vital staining with neutral red elected precisely the apical lip region as the osmophores in the three taxa. Light microscopical observations of sections of the labellum, in buds and flowers at anthesis, showed the glandular nature of the dome-shaped papillae that constitute the abaxial epidermis of the appendix in O. tenthredinifera . Besides these secretory cells, which possess a large nucleus, dense cytoplasm, numerous plastids and small vacuoles, and thin cuticle, the osmophore comprises also few layers of subepidermal parenchyma cells with abundant starch-rich plastids. The osmophore in O. bombyliflora , located in the fleshy appendix that forms a distinct concavity, has a structure roughly similar to that found in the osmophore of O. tenthredinifera , although the secretory activity is attributed to the adaxial rather than abaxial epidermis. Conversely, the osmophore in O. speculum seems to be comprised of the large domeshaped papillae of the broad lip border, together with several layers of subepidermal cells, but apparently without a detectable starch reserve. Histochemical tests allowed us to conclude that the osmophores synthesise a terpenoid-rich lipophilic secretion that seems to contain highly volatile compounds. These compounds may function as pollinators’ attractants at long distance, acting together with the previously reported low volatile cuticular hydrocarbons, in a multicomponent blend.
Euglossine bees and perfume orchids: what Darwin
1 didn’t know
Pemberton, RW 1 , Whitten, WM 1 , Ackerman, JD
Florida Museum of Natural History, USA; 2
2
University of Puerto Rico
Although Darwin was unable to observe euglossine bees and study their interactions with orchids, his far-flung correspondents sent descriptions of natural pollination, and his colleagues in England provided cultivated plants with which he studied the functional morphology of
Catasetum, Mormodes and Cycnoches flowers. He considered such examples as among the strongest in supporting his thesis that 'nature abhors self-fertilization' and that there must have been strong selection for such elaborated traits. Darwin was largely correct in his interpretation of floral morphology of euglossine bee pollinated plants, but he and his correspondents thought that the bees gnawed the flowers, not understanding the

significance of floral fragrance production to either the bees or the orchids. This extraordinary mutualism involves males of the ca. 190 species of Neotropical orchid bees (Apidae: Euglossini) and the ca. 700 perfume orchid species. Orchid bees are the exclusive pollinators of the orchids. Perfume orchids employ specialized floral fragrances composed of unique blends of compounds that attract particular bee species (often an array of species).
The bees collect these volatile compounds, store them in their spongy hind tibia, and then use these unmodified floral fragrances in their courtship rituals (sexual selection based on collected perfumes). Bees are manipulated into a position by both floral morphology and sites of scent production so that they remove the pollinarium, which becomes cemented to precise places on the bee’s body. The combination of floral fragrance filters and differential placement of pollinaria results in reproductive isolation among species of perfume orchids.
The mutualism is obligatory for the orchids but facultative for the bees, which can obtain their particular chemical needs from other sources such as aromatic leaves, bark, fungi, and the floral fragrances of other plants. Perfume orchid fragrances generally contain high concentrations of monoterpenes, sesquiterpenes, and simple aromatics. Male euglossine bee pollination has evolved independently in a number of lineages, both within the Orchidaceae and in other families (e.g.,
Araceae, Euphorbiaceae, Gesneriaceae). All species in some groups such as Catasetinae and Stanhopeinae are pollinated exclusively by male orchid bees that seek fragrances, whereas other groups (e.g., Oncidiinae,
Zygopetalinae and Lycastinae) contain perfume orchids and orchids that use other pollination systems. Speciation within these lineages is thought to occur when mutations cause alterations in fragrances resulting in a shift in the species of bees attracted to the flowers thereby facilitating reproductive isolation. Recent phylogenetic studies suggest that a shift to perfume-based euglossine bee pollination occurred via either reward or deceptive pollination systems. Reversals from male-euglossine bee pollination to other means of pollinator attraction are not known to have occurred. Life is good for perfume orchids, but how long will that last with the accelerating pace of local and global environmental change?
Darwin on the pollination of
Orchis
- what he taught us and what can we tell him today
Dafni, Amots 1 , Cozzolino, S 2
1 Haifa University, Israel; 2 Dept Structural and
Functional Biology, University Federico II of Naples,
Italy
The genus Orchis consists a considerable part in
Darwins's pioneering book The Various Contrivances by which Orchids are fertilized by insects (1862, 2nd edition
1877) which established the study of experimental orchid pollination biology. Not less than 10% of the book is devoted to the genus Orchis.
Darwin focused on the floral functional morphology of Orchis and its role to insure cross pollination, while considering the floral adaptations as a result of natural selection. Darwin was not aware of the deceptive nature of the flowers which have no nectar. He supposed, as did other contemporary biologists, that the visiting insects pierce the cell wall while looking for 'honey' and extract cell sap. Strangely enough, Darwin had never seen even one single insect as a pollinator under field conditions. The present paper will discuss the post-Darwinian developments in the following aspects: The taxonomic delineation of the genus Orchis s.l.; isolating mechanisms and hybridization in the genus; pollination syndromes and the genus, and Orchis s.l. as a model for testing hypotheses on evolution of deception.
Integrative phylogenetic evidence on the origin of island endemics in the Mediterranean region: comparisons between oceanic and continental
1 fragment islands
Conti, E 1
University of Zurich, Switzerland
Ever since Darwin’s seminal chapters on geographical distribution in The origin of species , islands have played a key role in the development of biogeographic theory.
One aspect that has not been sufficiently investigated concerns the influence of geologic origins (e.g., continental fragment vs. oceanic islands) on processes of colonization and speciation. Fragment islands, which were once part of continental plates, already harbored a full set of co-adapted communities when they separated from the mainland. Conversely, when oceanic islands emerged from the bottom of the ocean and for some time thereafter, they harbored either no or only a few species.
Fragment islands may be colonized via range expansion prior to plate separation, migration over temporary land bridges, or long distance dispersal following fragmentation, while oceanic islands may be colonized exclusively via long distance dispersal. The availability of open ecological niches also differs between fragment and oceanic islands, with an important temporal dimension attached to this factor. The Mediterranean
Region sensu lato (including the Macaronesian archipelago) provides an ideal setting to compare the influence of island origin on processes of colonization and speciation, because it includes both continental fragment (e.g., Corsica and Sardinia) and oceanic islands
(e.g, the Canarian archipelago). Additionally, recent improvements on molecular dating and ancestral area reconstruction methods now allow for a more effective integration of evidence from phylogenies with prior knowledge on past geologic (i.e., microplate movements, formation of temporary corridors, island formation) and climatic events (i.e., onset of the Mediterranean climate).
We focused our analyses on the origin of fragment and oceanic island endemics in Araceae, Boraginaceae, and
Rutaceae. The results supported the single colonization, followed by in situ diversification, of the Canarian archipelago by both Echium (Boraginaceae) and Ruta
(Rutaceae) during the Miocene, likely after the formation of the oldest island about 20 Mya and well before the onset of the Mediterranean climate in the Pliocene.
Additionally, the colonization of islands within the archipelago does not appear to conform to the classic
341

stepping-stone model in Ruta . The origin of Corso-
Sardinian endemics is likely explained by the fragmentation of the Hercynian mountain belt in the early
Oligocene for Helicodiceros muscivorus (Araceae), by the separation of the Corso-Sardinian microplate from the Apulian microplate in the middle Miocene for R. lamarmorae, R. corsica (Rutaceae), and A. pictum
(Araceae), by temporary land connections with neighboring landmasses during the Messinian Salinity
Crisis of the late Miocene for Borago , and by long distance dispersal during the Pliocene for Anchusa
(Boraginaceae). Altogether, our studies highlight the key role of both tectonic and eustatic processes of marine transgression-regression for the origin of endemics in the continental fragment islands of Corsica and Sardinia and the importance of identifying discrete time windows for colonization for the origin of endemics in the oceanic
Canarian archipelago.
Species distribution modeling: a predictive tool for island ecology and biogeography
Broennimann, O 1
1 University of Lausanne, Switzerland
Island ecosystems are among the most sensitive ecosystems to anthropogenic global change. Habitat change, over-exploitation and introductions of invasive species have heavily impacted island ecosystems in the past, and these impacts are projected to increase in the future. The ecological, geographical and evolutionary peculiarities of islands thus offer a unique opportunity to study the warning signs of global changes. In the last two decades, interest in species distribution models (SDMs) of plants and animals has grown dramatically. SDMs statistically relate the geographical distribution of species or communities to their present environment. Projections of these models onto the landscape allow deriving maps of probability of presence of species or communities in areas where they haven not been monitored. Recent advances in SDMs allow forecasting the anthropogenic effects of global change on species distributions at different spatial scales. SDMs thus represent a powerful tool to investigate the impact of global change in an insular context. Here, I first present recent advances in
SDMs, discuss the ecological principles and underpinning assumptions, and highlight critical limitations and decisions inherent in the calibration and evaluation of SDMs. Second, I review the use of SDMs for the study of island ecology and biogeography in the literature, such as patterns of endemism and adaptive radiation, impacts of biological invasions, climate change and habitat destruction, restoration programs and design of nature preserves. I finally underline new topics of research for which SDMs could provide valuable insights.
Assessing the General Dynamic Model of island biogeography with a process-based approach based on plant species
Sarmento Cabral, J 1 , Wiegand, K 2 , Kreft, H 1
1 Biodiversity, Macroecology and Conservation
Biogeography, University of Göttingen, Germany;
2 Ecosystem Modeling, University of Göttingen, Germany
342
Recently, Whittaker and colleagues proposed a General
Dynamic Model (GDM) expanding island biogeography theory by including island age and geology. This made the theory dynamic and the authors made several predictions under this new framework. We addressed core GDM predictions by simulating island biogeographic and demographic processes of insular plant species on islands through geologic time. We increased and decreased island size for mimicking island emergence and expansion from volcanic activity and posterior erosion. We investigated islands with different degrees of isolation, which was varied either by distance to the mainland or by lower dispersal abilities of mainland species pools. We simulated long-distance
(from mainland) and intra-island dispersal, germination, growth, mortality, reproduction, speciation and extinction. Species competed for space and varied in size and in demographic properties, following metabolic trade-offs. Results revealed that individual colonization history and flora composition of the source mainland greatly influenced the fate of species richness, speciation and community assembly over geologic time.
Nevertheless, general trends could be identified and compared to the GDM. Similar to GDM predictions, species richness was a hump-shaped function of island age, being highest for less isolated islands. At advanced ages, richness tended towards a saturation level that was common to all islands, even with different isolation degrees. This suggests that the island species carrying capacity is gradually been reached, mostly due to accumulation of endemic species. Moreover, speciation rate and endemic species richness were humped-shaped, with increasing number of endemic diversifying lineages
(species originating from a common colonizing ancestor) towards island maturity. In contrast to GDM predictions, immigration and extinction curves were hump-shaped, with island size playing a major role than island age.
However, the GDM predictions still partially held: there were more immigration events on younger islands and more extinction events for older islands. Furthermore, less isolated islands had a higher absolute number of diversifying lineages. This can be explained by a larger number of colonizing species, and, thus, more potentially diversifying colonizers. Interestingly, more isolated islands produced more species per diversifying lineage, showing a higher potential for adaptive radiation than less isolated islands. Also in opposition to GDM predictions, both proportional endemism and species number per diversifying lineage increased with island age, probably because endemics were better adapted than non-endemic species and, therefore, were better able to survive island erosion and random disturbances. Results indicate the usefulness of process-based models for simulating species richness and enhancing biogeographic theory with a mechanistic understanding on processes influencing plant species richness.
Getting the right measure of island isolation – a global test of insular floras
Weigelt, P 1 , Kreft, H 1
1 Biodiversity, Macroecology and Conservation
Biogeography Group, University of Göttingen, Germany
According to island biogeography theory, isolation is a core driver of species richness on islands. So far most

studies have investigated the effect of isolation measured as the ordinary distance to the nearest continent hypothesizing that mainlands are the most important sources for colonization. We compared the explanatory power of eight fundamentally different types of isolation metrics in almost 100 variations on vascular plant species richness of 453 islands at global scale. We considered the distance to the nearest mainland, distance to other islands, stepping stones, the proportion and distance of neighboring landmass, prevailing wind and sea current directions as well as climatic similarity between source and target areas. The applied isolation metrics represent different mechanisms like long distance dispersal, island hopping and metapopulation effects as influential components of colonization and maintenance of plant richness. The distance to the nearest mainland explained
25 % of the variation. Stepping stone distances, distances to large islands and the proportion of neighboring landmass gained equally high predictive power or yet performed slightly better. In a multi-predictor model accounting for area, climate, topography, and island geology, stepping stone distances to source landmasses of at least 100,000 km² as well as the proportion of neighboring landmass gained strongest support.
Including both metrics significantly improved model fits compared to a model including the ordinary distance to the mainland. The full model explained 82 % of the variation. Our results show that different colonization mechanisms influence plant species richness on islands.
We demonstrate that such divergent mechanisms can be found even at macro-scales. However, due to multicollinearity among the isolation metrics it is difficult to disentangle their relative importance. Due to small statistical differences, we conclude that the ordinary distance between an island and the nearest mainland serves as an adequate and simple-to-calculate measure of island isolation even at global scale.
The influence of environmental spatial structure on the predictability of plant community composition
Super, L 1 , Pither, J 1
1 University of British Columbia, Vancouver, Canada
Ecologists agree that plant communities are positioned at different points along a continuum of stochastic to deterministic community assembly. Identifying the factors that govern the positioning of communities along this continuum represents an important challenge for academics and practitioners alike. Empirical and theoretical research underscores the importance of understanding how species’ niche characteristics, their dispersal characteristics, and environmental heterogeneity interact to govern the balance between deterministic and stochastic assembly. However, little attention has been paid to the potentially unique influences of spatial-environmental structure. In particular, no study to our knowledge has examined if and how varying the degree of spatial clustering of the environment within a landscape influences the importance of deterministic relative to stochastic assembly within local plant communities. We addressed this question using spatially-explicit simulations of competitive, source-sink metacommunities. We conducted a fully replicated, factorial experiment that crossed environmental clustering (random to positive spatial autocorrelation), environmental composition
(uniform vs. Gaussian distribution of environment amounts), fundamental niche scenario (neutral vs. species-packing model), and dispersal capacity (local vs. global). We quantified variation in the importance of stochastic relative to deterministic assembly by calculating the numbers equivalents of Shannon beta diversity among replicate communities. In the absence of either niche differentiation or dispersal limitation, spatial clustering in the environment had no impact on the predictability of local community composition. However, in the presence of both dispersal limitation and niche differentiation (representative of real plant communities), spatial clustering of the environment significantly increased the predictability of community composition
(i.e. decreased the numbers equivalents for Shannon beta diversity). This effect was apparent even when the amount of optimal habitat in the landscape was equal for all species in the metacommunity. These findings indicate that the current emphasis on the interaction between environmental heterogeneity, niche differentiation, and dispersal limitation needs to be refined to include spatial configuration of environmental heterogeneity. From a practical standpoint, our findings suggest that conservationists and restoration ecologists would benefit from considering the spatial configurations of environments when conserving plant communities.
Multiple colonizations from Madagascar and converged acquisition of dioecy in the Mascarene
Dombeyoideae (Malvaceae) as inferred from chloroplast and nuclear DNA sequence analyses
Le Péchon, T 1 , Dubuisson, J-Y 2
1
Gigord, L 4
France;
, Haevermans, T
Université Pierre et Marie Curie, Paris,
Muséum National d’Histoire Naturelle, Paris,
4 Conservatoire Botanique National de
Mascarin, Saint Leu, La Réunion, France
3 ,
Université de La Réunion, Sainte Clotilde, La Réunion,
France;
France; 3
2
In the Mascarenes, a young oceanic archipelago composed of three main islands, the Dombeyoideae
(Malvaceae) have diversified extensively with a high endemism rate. With the exception of the genus
Trochetia , Mascarene Dombeyoideae are described as dioecious whereas Malagasy and African species are considered to be monocline, species with individuals bearing hermaphrodite/perfect flowers. In this study, the phylogenetic relationships were reconstructed to clarify the taxonomy, understand the phylogeographic pattern of relationships and infer the evolution of the breeding systems for the Mascarenes Dombeyoideae. Parsimony and Bayesian analysis of four DNA markers (ITS, rpl16 intron and two intergenic spacers trn Qrsp 16 and psb Mtrn D) was used. The molecular matrix comprised 2985 characters and 48 taxa. The Bayesian phylogeny was used to infer phylogeographical hypotheses and the evolution of breeding systems. Parsimony and Bayesian trees produced similar results. The Dombeyoideae from the Mascarenes are polyphyletic and distributed among four clades. Species of Dombeya , Trochetia and Ruizia are nested in the same clade, which implies the paraphyly of Dombeya . Additionally, it is shown that each of the four clades has an independent Malagasy origin. Two adaptive radiation events have occurred within two
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endemic lineages of the Mascarenes. The polyphyly of the Mascarene Dombeyoideae suggests at least three independent acquisitions of dioecy. This molecular phylogeny highlights the taxonomic issues within the
Dombeyoideae. Indeed, the limits and distinctions of the genera Dombeya , Trochetia and Ruizia should be reconsidered. The close phylogeographic relationships between the flora of the Mascarenes and Madagascar are confirmed. Despite their independent origins and a distinct evolutionary history, each endemic clade has developed a different breeding systems (dioecy) compared with the Malagasy Dombeyoideae. Sex separation appears as an evolutionary convergence and may be the consequence of selective pressures particular to insular environments.
1
Miocene dispersal drives island radiations in
Trachycarpeae (Arecaceae/Palmae)
Bacon, CD 1 , Baker, WJ 2 , Simmons, MP 1
Colorado State University, Fort Collins, USA; 2 Royal
Botanic Gardens, Kew, UK
In an effort to elucidate patterns of island radiation the phylogeny of tribe Trachycarpeae (Arecaceae/Palmae; ca. 269 species in 18 genera) was inferred from three chloroplast ( mat K, ndh F, and trn DT) and three low-copy nuclear genes (CISPs 4 and 5 and RPB2). The resulting phylogenetic trees, which were generally well resolved and highly supported, indicate the need for taxonomic changes, including the resurrection of Saribus . Historical biogeography analyses were conducted using a likelihood
(dispersal–extinction cladogenesis model) approach that inferred the Trachycarpeae to be of southern North
American, Central American, or Caribbean origin. The mean age of the tribe under a relaxed lognormal uncorrelated molecular clock assumption was estimated to be 37.74 Ma. The relationships between and within genera were interpreted with respect to divergence times and major geological and climatic factors. Events inferred to have influenced extant and disjunct distributions are described, particularly the GAARlandia and North Atlantic land bridges, tectonic movement in
Southeast Asia, and climatic shifts between the Eocene and Pliocene. When considering the three major island radiations within the Trachycarpeae, Miocene dispersal appears to have been the driving force in causing allopatric speciation and is further identified as an emerging pattern across the tree of life.
Genetic consequences of anagenesis in oceanic island plants
Takayama, K 1 , Sun, B-Y 2 , Stuessy, TF 1
1 Institute of Botany, University of Vienna, Vienna,
Austria; 2 Faculty of Biological Sciences, Chonbuk
National University, Jeonju, Korea
Oceanic islands are valuable as natural laboratories of plant evolution, and they have long fascinated evolutionary biologists. In particular, numerous investigations have focused on processes of speciation in island endemics. The most commonly described model of speciation on oceanic islands has been
 gcladogenesis' (also known as adaptive radiation in an
344 island context). From a single ancestor by selection within markedly different ecological zones, several lines of speciation develop rapidly. Morphological and physiological divergence among species in this model is often dramatic, but overall genetic differentiation may be relatively low. A contrasting model of speciation is
‘anagenesis’ (also known as simple geographic or phyletic speciation), whereby an initial founder population simply diverges through time without further specific differentiation. Anagenesis may be much more important in oceanic islands than previously believed, accounting for levels of endemic specific diversity from
7% to 88% (with a mean for all islands of 25%, as inferred from published floras of oceanic island plants).
In particular, low-elevation islands with low habitat heterogeneity, such as Ullung Island (88%), Ogasawara
Islands (53%), and St. Helena (53%), are interpreted to reflect higher levels of anagenesis. These two different modes of speciation not only lead to very different levels of specific diversity, but they also would be expected to yield different genetic consequences. Earlier molecular studies in Ullung Island, Korea, revealed high levels of genetic variation in the endemic species, Acer takesimense and Dystaenia takesimana , which are regarded as having been anagenetically derived.
Furthermore, there is no geographic partitioning of genetic variation within the island species; each behaves as one large population. Anagenetically derived species, therefore, may accumulate genetic variation through time and present a very different genetic pattern from those that have resulted from cladogenesis (adaptive radiation).
Only a few attempts, however, have so far examined critically the genetic consequences of anagenesis in contrast to the classical model of cladogenesis. In this study, we examine patterns of genetic variation in Acer okamotoanum and A. takesimense , using nuclear microsatellite markers, in comparison with their continental progenitors. A synthesis of information from molecular studies in the Ogasawara (Bonin) Islands is also provided. These two island systems, Ullung and
Ogasawara, are ideal for studying anagenesis among endemic species, and they permit evaluation of genetic consequences of anagenetic vs. cladogenetic speciation in oceanic islands.
Global diversity of island floras: species richness, endemism, and threat
Kreft, H 1
1 University of Göttingen, Germany
Island research has prompted some of the most influential theories in ecology, biogeography and evolution. Islands continue to be popular model systems but 200 years of island research have produced only a sketchy understanding of the global-scale distribution of vascular plant diversity on islands. This is particularly glaring given the multiple threats island floras are facing in an increasingly globalized world. Moreover, environmental and historical correlates of diversity as well their underlying mechanism are to date heavily debated. Here, I review recent progress in assembling a global-scale picture of island plant diversity. I demonstrate how plant species richness can be reasonably well predicted from a few core island characteristics (area, isolation, temperature, precipitation,

topography, geology). A multi-predictor model comprising of these six variables accounts for 85% of the variation in a global set of 488 islands also accounting for the overall lower richness of island floras and reconciling classic equilibrium theory with modern advances in macroecology. In a second step, we looked at endemism richness – a measure that combines endemism and species richness – and current and potential future threats of island and mainland floras.
Despite their overall lower species densities, islands have a significantly higher concentration of species’ ranges and contribute significantly to the world’s biodiversity.
These analyses reveal that island biotas are currently more heavily threatened by human activities than mainland regions. Future projections of land use change suggest that habitat loss might even accelerate on islands as compared to mainland areas. These results clearly suggest that islands should have a top priority in global conservation efforts. I conclude this talk by providing an outlook on how island biogeography can make progress by compiling existing but scattered floristic information and by tapping into recent developments in the availability of digital geographic information as well as statistical and process-based mechanistic modeling.
The endemic flora of Micronesia: implications for conservation in the Pacific and beyond
Costion, C 1 , Lorence, D 2 , Kitalong, A 3
1 Australian Tropical Herbarium/University of Adelaide,
2
3
Australia; National Tropical Botanic Garden, USA;
Belau National Museum, Republic of Palau
Micronesia is part of one of the world's top biodiversity hotspots, the Polynesia–Micronesia hotspot. Biodiversity hotspots have been selected based on number of endemic species and degree of threat. We present an up to date and complete checklist of endemic vascular plants for the entire Micronesia bioregion. Previous estimates have been substantially revised. We emphasize the importance of conducting similar regional works in the Pacific region to prioritize conservation efforts and suggest areas where this is needed or taxonomy is out of date. Once endemic status is determined for a species, then it can be prioritized for Red List assessments. These can be difficult on tropical oceanic islands where there are few collection records and poor distribution data. We outline a strategy to overcome this limitation using the pollen record from palaeo-environmental studies in the Republic of Palau and propose that similar approaches can be used on other islands that are rich in poorly known plant species.
A multigene phylogeny of liverworts
Shaw, J
Von Konrat, M 3 , Engel, J 3
L 6
1 , Shaw, B 1
, Devos, L 1
, Crandall-Stotler, B 2
, Long, D 5
, Stotler, R 2
, Davis, C 4
,
, Forrest,
1 Duke University, Durham, USA;
University, USA; 3
2 Southern Illinois
Field Museum, Illinois, USA;
Technical Community College, USA;
4 Durham
5 Edinburgh
University, UK; 6 University of Connecticut, USA
Phylogenetic relationships among the liverworts were reconstructed based on nucleotide sequences from eight nuclear, mitochondrial, and plastid loci. The complete data matrix consists of approximately 1200 accessions representing the major clades and gametophyte body plans (complex thalloid, simple thalloid, leafy) found among liverworts. Because the liverworts are one of, if not the earliest clade of land plants, our reconstruction provides a unique insight into early embryophyte morphological evolution. The phylogeny also provides the framework for additional investigations of biogeographic patterns, reproductive biology, morphology, developmental biology, and genome evolution. We combine the liverwort data set with comparable data from the mosses, another diverse lineage of early, prevascular embryophytes, to compare the timing of diversification in these two clades.
A molecular phylogenetic approach to resolving the
Lepidozia
generic complex
Cooper, E 1,2 , Henwood, M 1 , Brown, E 2
1 School of Biological Sciences, University of Sydney,
Australia; 2 National Herbarium of New South Wales,
Sydney, Australia
Telaranea as traditionally circumscribed is known not to be monophyletic. Species which possess leaves with a distinct disc are more closely related to Lepidozia than they are to those species whose leaves consist only of uniseriate lobes. However, relationships within this
Lepidozia – Telaranea lineage are unresolved. We constructed a phylogeny of Lepidozia and Telaranea using nucleotide sequences from one mitochondrial
( nad 5nad 4), two nuclear (nrITS1 and nrITS2) and two chloroplast ( trn L-F and rbc L) loci. In order to identify clade synapomorphies, we mapped morphological characters onto the phylogeny. We used Bayesian relaxed clock dating methods with fossil constraints and a 150 genus outgroup spanning the Marchantiophyta, to estimate divergence times. The Lepidozia – Telaranea lineage originated in the early Cretaceous during a period of rapid diversification within the Lepidoziaceae. The earliest divergent lineage is comprised of Telaranea species which have waxy cuticular elaborations giving the plants a glaucous appearance. By the mid-Cretaceous a second lineage of Telaranea species had diverged.
Within this lineage, there is a pattern of progressive reduction in the area of the leaf disk and elongation of the uniseriate portion of the lobes, possibly as a result of adaptive radiation within high moisture micro-habitats.
The next divergence occurred early in the paleogene and separated Lepidozia from the remainder of Telaranea .
This was accompanied by a shift towards leaf asymmetry and loss of a differentiated stem epidermal cell layer in
Lepidozia . We find evidence for recent speciation within each of these four lineages. Telaranea species which lack a leaf disc and have reduced stems belong to three other lineages and not the Lepidozia – Telaranea clade. The type species of Telaranea , T. chaetophylla , has this reduced stem and leaf anatomy and does not belong to
345

the Lepidozia – Telaranea clade. Three new genera will require description to reconcile the taxonomy with the phylogeny.
Biogeographic insights from phylogenetic analyses of liverwort lineages
Heinrichs, J 1 , Feldberg, K 1
Von Konrat, M 3
, Kreier, H-P 1 , Schmidt, AR 2 ,
1 Dept of Systematic Botany, Georg-August-University
Göttingen, Germany; 2 Courant Research Centre
Geobiology, Georg-August-University Göttingen,
Germany; 3 Field Museum, Dept of Botany, Chicago,
USA
In the 19th and early 20th century, bryologists preferably used a geographical or typological species concept where species were defined as largely invariant units. Many liverwort taxa were known only from type material and were classified as species with narrow distribution ranges. More recently, authors accepted intraspecific morphological variation and lowered numerous local taxa to synonyms of widespread species. These species often colonized several continents. Disjunct ranges of liverworts have frequently been explained by ancient vicariance and slow rates of morphological evolution.
However, molecular data led to new appraisals of distribution ranges and allowed the reconstruction of refugia and migration routes. We show that intercontinental ranges of bryophytes were often caused by dispersal rather than geographical vicariance. A great deal of distribution patterns of disjunct bryophytes were likely formed by processes such as short distance dispersal, occasional long distance dispersal, extinction, recolonization and diversification. Many morphologically circumscribed liverwort species comprise several morphologically cryptic molecular lineages. Oftentimes these lineages reflect a geographical pattern, pointing to local diversification events and contradicting a general panmixis hypothesis. There is growing evidence that liverworts date back to the
Ordovician and represent the most ancient lineage of extant land plants. Divergence time estimates based on
DNA sequence variation point to an origin of the main liverwort lineages in the late Palaeozoic and diversification of most liverwort crown groups in the late
Mesozoic and Cenozoic; however, detailed estimates are hampered by the sparse fossil record of bryophytes. We present new fossils that improve our knowledge of the time span of liverwort evolution and discuss the relevance of a dense fossil record for reliable reconstructions of divergence times.
Morphological evolution in hepatics – patterns, trends and mechanisms
Crandall-Stotler, B 1
Renzaglia, K 1
, Bartholomew-Began, S 2 ,
1 Southern Illinois University at Carbondale, USA;
Chester University, Pennsylvania, USA
2 West
Long before molecular phylogenetics resolved liverworts as the sister group to the rest of the embryophytes, morphologists considered liverworts to be pivotal in early land plant evolution, primarily because they exhibit
346 both thalloid and leafy shoot morphologies and they have the least complex sporophytes of the embryophytes.
Proponents of the antithetic theory of embryophyte origin hypothesized progressive evolution within hepatics from small, thalloid forms like Riccia to large, leafy plants like
Haplomitrium , while adherents of the homologous theory proposed the opposite scenario. In contrast to the intuitive approaches of the past, the comprehensive molecular phylogeny of the Liverwort Tree of Life provides a framework upon which patterns and trends in the evolution of liverwort diversity can be reconstructed.
To evaluate evolutionary patterns across all levels of the tree, we have partitioned the morphological data, including ultrastructural, ontogenetic and anatomical characters, into three nested matrices, one focused on resolving trends at the deepest nodes of the tree and the others, at the intermediate and terminal nodes, respectively. Ancestral state reconstructions generated in our analyses show that there is an overall tendency for reduction in size and complexity of both sporophytes and gametophytes from the first divergences to the crown group lineages in most of the major clades, as predicted by adherents of the homologous theory of hepatic evolution. Transformations in primary protonemal form, mature apical cell geometry, mature sperm cell organization, gametangial division patterns, and form of the prophase I sporocytes appear to have occurred at the deep nodes that define the five backbone clades. For example, different sperm cell architectures are consistently diagnostic of the Haplomitriopsida,
Marchantiopsida and Jungermanniopsida clades, and transformations from tetrahedral to cuneate and tetrahedral to lenticular apical cell geometries characterize the divergences between the
Haplomitriopsida and Marchantiopsida, and the
Jungermanniidae and Metzgeriidae clades, respectively.
Transformational reversions and reductive character loss result in substantial homoplasy and a lack of polarization for most anatomical characters across the entire tree, although some recurrent patterns were identified within specific clades. For example, in several lineages of leafy liverworts isophylly is reconstructed as derived from anisophylly and stem perigynia, from shoot calyptrae and in complex thalloid liverworts there is a trend for elaborate carpocephala to be reduced or lost in several lineages. New studies of spore germination patterns, protonemal ontogenies and the developmental transitions from juvenile to adult gametophyte morphologies have aided in identifying structural homologies among pivotal taxa and have verified that paedomorphosis is a significant evolutionary mechanism in hepatics.
Liverworts are unique among land plants in their extended matrotrophy and internalization of the sporophyte generation, a life strategy that has doubtless impacted their morphological evolution. Thus, although knowledge of the morphological, ultrastructural and ontogenetic patterns and processes of the earliest diverging liverwort taxa is relevant to modeling early land plant evolution, how evolution has proceeded at intermediate and terminal levels likely is not.
Chloroplast and mitochondrial genome evolution in early land plants: inferences from assemblies of liverwort genomes
Goffinet, B
Wang, B 3
1 , Liu, Y 2 , Forrest, L 1
, Villarreal, JC 1 , Li, L 2
, Xue, J
, Qiu, Y-L 2
3 , Wickett, N 3 ,

1 Dept of Ecology and Evolutionary Biology, University of Connecticut, USA; 2 Dept of Ecology & Evolutionary
Biology, University of Michigan, Ann Arbor, USA;
3 School of Life Sciences, Nanjing University; Penn State,
USA
Chloroplasts and mitochondria harbor a distinct set of genes assembled into a single putatively circular chromosome. The composition of these genomes has been dramatically altered since the endosymbiotic acquisition of a prokaryotic cell. In particular, the genome has undergone a significant reduction in size, through the loss of genes that are redundant with the
'host's' nuclear genes or not required for the persistence of these organelles, or through the transfer of the genes to the nuclear chromosomes. By the time the
Charophycaeae gave rise to the ancestor to land plants about 500 mya, the chloroplast genome (i.e., plastome) was reduced to about 130 kb and ± 120 genes, arranged into a large and single copy region separated by two inverted repeats. During the diversification of land plants, the size of the plastome remained little altered as gene losses were rare except in lineages characterized by dramatic shifts in life histories (i.e, parasitic plants).
Changes in gene order are more frequent and in particular the length of the inverted repeats varies among lineages, at least among vascular plants. Plastome architecture is rather highly conserved among bryophytes based on 6 currently available genomes, including two liverworts. We have sampled complete plastid genomes, using assemblies of reads bioinformatically extracted from whole genome shotgun 454 sequencing, for nine generic exemplars [ Blasia , Haplomitrium ,
Fossombronia , Metzgeria , Moerkcia , Pleurozia ,
Ptilidium , Scapania and Treubia ] spanning the liverwort tree of life. A survey of the genome composition and structure reveals that only two genes (cysA and cysT) may be lacking form the plastome of some – distantly related – lineages and more importantly that the plastid genomes of all photosynthetic liverworts are colinear.
Rare gene losses, large inversions and changes in the boundaries of the inverted repeat as seen in mosses and hornworts, respectively, are absent. Compared to vascular plants plastomes, the cp genomes of bryophytes, and in particular liverworts are thus extremely stable in terms of their size, composition and organization. Plant mitochondrial genomes have been known for a long time to be highly unusual in both genome structure and sequence evolution. However, only two early land plant
(one liverwort and one moss) mitochondrial genomes have been sequenced, and it is unknown when the plastome acquired all the unusual features during its evolution in green plants. We have sequenced mitochondrial genomes from two liverworts ( Treubia lacunosa and Pleurozia purpurea ), one moss ( Anomodon rugelii ), and two hornworts ( Megaceros aenigmaticus and Phaeoceros laevis ). A comparative analysis of these seven genomes shows that the gene order and sequence evolve slowly, but intron gains, gene losses, RNA editing, and genome size changes are very active.
Overall, chondriome exhibits a mixed mode of conservative yet dynamic evolution in early land plants.
The highly dynamic genome structure and sequence evolution seen in angiosperms did not begin until the origin of vascular plants.
Pattern and mode of speciation of New Caledonian
Diospyros
(Ebenaceae)
Samuel, R 1 , Turner, B
Wallnoefer, B 4
1 , Duangjai, S 2 , Munzinger, J 3
, Chase, M 5
,
1 Dept of Systematic and Evolutionary Botany, University of Vienna, Austria; 2 Dept of Forest Biology, Kasetsart
3 Laboratoire de
4
University, Bangkok, Thailand;
Botanique, Centre IRD de Nouméa, New Caledonia;
Natural History Museum, Vienna, Austria;
Laboratory, Royal Botanic Gardens, Kew, UK
5 Jodrell
Diospyros is one of the largest genera of angiosperms, comprising approximately 500 species of which 31 are distributed in New Caledonia. Of these species all except
D. fasciculosa are endemic to New Caledonia. Recent molecular studies on family Ebenaceae where a subset of
New Caledonian Diospyros was included highlighted the presence of four lineages and two types of endemism.
The first being paleoendemics, which suggested ancient
Gondwana (Australian) origin, e.g., D. macrocarpa, D. brassica, D. balansae , and the second neoendemics, recent Indo–Malesian elements that include species like
D. vieillardi, D. umbrosa, D. parviflora etc. Species boundaries among most of the neoendemics seems to be unclear and are not well accepted by all authors. The ongoing project on New Caledonian Diospyros uses
AFLP analysis to determine species boundaries (i.e. taxonomic units) of the neoendemics. Molecular phylogenetics using rapidly evolving plastid and lowcopy nuclear sequences will detect hybridization and introgression that could have given rise to speciation as well as reproductive isolation that has evolved as a consequence of divergent selection on traits in different environments and thus ecological speciation. Speciation is often accompanied by chromosomal rearrangment both numerical and structural, which will be investigated for the New Caledonian Diospyros .
Phylogenetic and biogeographic relationships within
1 the endemic subtribe Espeletiinae Cuatrec.
(Asteraceae) of the South American páramos
Diazgranados, M 1, 2 , Barber, JC 1
Saint Louis University, USA; 2 Missouri Botanical
Garden, St Louis, USA
The high elevation grassland ecosystem of the Northern
Andes, known as páramo, is the most diverse ecosystem of the high elevations of the world, and is an ideal system to understand rapid adaptive radiations and speciation mechanisms in sky islands. The primary goal of this project is to understand the patterns and mechanisms of speciation in the páramos, and the consequences of climate change on the geographic distribution of species.
This is being accomplished using the dramatic radiation of frailejones (subtribe Espeletiinae, family Asteraceae) of the Andean forest and páramos, which provide a potential model system for investigating these phenomena. The nearly 143 named species are grouped into eight genera: Carramboa (5 spp.), Coespeletia (6
347

spp.), Espeletia (68 spp.), Espeletiopsis (27 spp.),
Libanothamnus (11 spp.), Paramiflos (1 sp.), Ruilopezia
(24 spp.) and Tamania (1 sp.). These plants are widely distributed and abundant in the high Andean forest and páramos of Colombia, Venezuela and, to a lesser extent,
Ecuador, and the preservation of the páramo ecosystem depends largely on them. Although there have been some previous attempts to reconstruct the evolutionary history of this group based on morphological and molecular data, relationships among genera and species remain largely unresolved. This project aims to answer the following questions: 1) what are the phylogenetic relationships among species and genera of the subtribe Espeletiinae?
2) is there any correlation between the phylogeny and the phytogeographic relationships within the subtribe? and 3) will climate change affect significantly the geographic distribution of species of the subtribe? We conducted extensive fieldwork covering most of the páramos of the northern Andes, with special emphasis on unexplored or uncollected areas. We found 13 probable new species
(one of Coespeletia , eight of Espeletia , three of
Espeletiopsis , and one of Libanothamnus ), the first report of the genus Ruilopezia for Colombia, several new reports for localities, numerous putative hybrids, a few species previously thought to be extinct, and identified several critically endangered species. Phylogenetic analyses were based on multiple markers: sequence data from nrDNA (ITS and ETS), AFLP (18 loci) and microsatellites (10 loci). Results show strong support for the monophyly of the subtribe. Relationships reflect a geographic structure, with one clade containing only
Venezuelan species and another comprising only
Colombian species, and several internal clades representing geographic complexes. None of the genera as currently described is monophyletic, and the position of Carramboa supports the proposed origin of the subtribe in Venezuela. Incongruence between the different data sets may be due to hybridization, incomplete lineage sorting, and/or differential tempo in evolution of the markers. Biogeographic analyses show no statistical correlation between species radiations and habitat diversity, suggesting that these species are restricted to specific habitats (i.e. niche conservatism).
Comparison of modeled niches under future and current scenarios of climate show significant differences, with areas being on average smaller in the future. This suggests that in the short term, climate change will have a critical impact on the distribution and conservation of these species.
Phylogenetic relationships of
Muehlenbeckia
and the new Australian genus
Duma
based on chloroplast and nuclear sequence data
Schuster, TM 1 , Wilson, KL 2 , Kron, KA 1
1
2
Wake Forest University, North Carolina, USA;
RoyalBotanic Gardens and Domain Trust, Sydney,
Australia
Muehlenbeckia is a group in Polygonaceae that includes lianas, vines and scrambling or erect (sub)shrubs. The group occurs only in the Southern Hemisphere and has a disjunct distribution pattern. Most species occur in
Australasia and nine species are found in Central and
South America. Muehlenbeckia presently includes 31 species and is one of several genera previously
348 segregated from a highly variable Polygonum s . l . The monophyly of Muehlenbeckia and its evolutionary relationships to other genera in subfamily Polygonoideae are unclear. Muehlenbeckia ’s traditional placement in
Coccolobeae with Antigonon , Brunnichia , and
Coccoloba that were shown to be members of
Eriogonoideae was not supported in previous studies.
Instead, Muehlenbeckia is more closely related to
Fallopia in tribe Polygoneae. Twenty-two species of
Muehlenbeckia and representatives of related genera including Atraphaxis , Fallopia (including Reynoutria ) and Polygonum s .
s . were sampled for this study.
Molecular data were generated from five chloroplast
( mat K, ndh F, 3’ rps 16-5’trn K, trn Ltrn F and 3’ trn Vndh C) and two nuclear gene regions (second intron of
LEAFY , ITS). Analyses were done using Maximum
Likelihood and Maximum Parsimony methods. The results show that tribe Polygoneae is supported as monophyletic and is composed of (in branching order from the deepest node): Knorringia , Atraphaxis ,
Polygonum s .
s . and Polygonella , the new genus Duma ,
Reynoutria , Fallopia and Muehlenbeckia . Three
Australian species of Muehlenbeckia fall outside of the main clade and are sister to Reynoutria and Fallopia +
Muehlenbeckia , a relationship that is supported with strong bootstrap. This result is surprising because no previous workers questioned the inclusion of these three species in Muehlenbeckia . Based on the findings of this study and because there was no legitimate generic name for this group, Muehlenbeckia coccoloboides , M . florulenta and M . horrida received the new genus name
Duma T.M.Schuster. Duma is a group of spinescent shrubs that do not possess extrafloral nectaries (the latter are present in Muehlenbeckia ). The stems of Duma are glaucous or have a striking white–cream layer of tissue and the older bark is grey and corky. They have linear, subsessile leaves that may be deciduous at maturity.
Flowers appear in clusters at the nodes. Duma coccoloboides occurs in the desert region of South
Australia and the widespread D . florulenta grows in seasonally flooded areas. Duma horrida occurs in restricted populations in both Eastern and Western
Australia and is associated with seasonally dry lakes and water courses. The latter two species are tolerant of high levels of salinity. Neither Muehlenbeckia nor Fallopia is monophyletic, with the main group of Muehlenbeckia most closely related to two successive clades of Fallopia .
One clade of Fallopia , previously characterized as
Reynoutria , is sister to F . section Fallopia +
Muehlenbeckia . Extrafloral pit nectaries that are found at the base of the articulated petiole are a synapomorphy for
Reynoutria + Fallopia + Muehlenbeckia . Relationships in
Fallopia and Reynoutria correlate with base chromosome numbers, while relationships within Muehlenbeckia correspond to geographic distribution patterns. Within the Muehlenbeckia clade, there are three subclades that generally correspond to groups formed by species either from Australia, New Zealand or Central and South
America.
1
Biogeography and phylogenetic relationships in
Cryptolepis
R.Br. (Periplocoideae, Apocynaceae)
Joubert, L 1 , Bruyns, P 2 , Klak, C 2 , Venter, A 1
University of The Free State, South Africa; 2 University of Cape Town, South Africa

Cryptolepis R.Br., consisting of 32 species, is the second largest genus in the Periplocoideae (Apocynaceae). It is also one of the most widely distributed genera of the subfamily, occurring throughout sub-Saharan Africa, in
Socotra and Yemen in Arabia and in East Asia in India and China. The species of Cryptolepis occur in a wide range of habitats such as semi-desert, savannah and forest and range from trees to climbers or small, almost spiky shrubs. Some of the species are morphologically easily definable and are restricted to specific habitats, while others are morphologically highly variable and occur in a broad spectrum of habitats. We use the loci nuclear ITS and chloroplast trn D-T and trn T-F to reconstruct the phylogeny of Cryptolepis . With this we investigate the relationships among the species and the monophyly of some of the morphologically more variable species.
Allelic variation of floral isolation genes in a sexually deceptive orchid
Xu, S 1,2 , Schlüeter, P 1 , Schiestl, F 1
1 Institute for Systematic Botany, University of Zürich,
2 Switzerland; Institute of Integrative Biology ETH
Zürich, Switzerland
In plants, pollinator mediated pre-mating isolation (floral isolation) can play an important role in the speciation process. Ophrys is a genus of sexually deceptive orchids, which mimics the mating signals of female pollinators
(mainly sex pheromones). These orchids thereby induce so-called pseudocopulations in their male pollinators, leading to pollination. Pollinator attraction in Ophrys is very specific, since the mating signals in pollinators are specific. Differential pollinator attractions can act as premating isolation among closely related species. This has been confirmed by in situ pollen tracking on closely related sympatric species of Ophrys . Further analysis suggests that the floral isolation is mainly due to different alkenes produced by different species. Thus, the genes responsible for differential alkene production among species may be floral isolation genes. Member of the stearoyl-acyl-carrier-protein desaturase (SAD) gene family may be involved in alkene production in Ophrys .
Allelic gene expression and evolutionary patterns of
SAD gene family members were investigated among different species on the population level. We found three
SAD homologues, which showed strong directional selection on allelic expression levels and / or loss of function among different species. Our data is consistent with a scenario in which speciation in sexually deceptive
Ophrys orchids may happen rapidly and possibly in sympatry due to changes at few loci of large effect in the genome.
To hide or not to hide: the influence of apparency on the pollination and herbivory of an endangered terrestrial orchid
Faast, R 1 , Facelli, JM 1 , Austin, AD 2
1 School of Earth and Environmental Sciences, University of Adelaide, Australia; 2 Australian Centre for
Evolutionary Biology and Biodiversity, University of
Adelaide, Australia
The ability to attract pollinators is essential for the reproductive success of plants relying on animals for ovule fertilization. As such, the ease with which plants are located, or their apparency, can be an important determinant of pollination success. Many plants have evolved traits that optimise their attractiveness, such as colour, size or stature, or the number of flowers produced. These intrinsic traits are often genetically controlled and can be subject to pollinator-mediated selection. However, characteristics of the biotic community can also play a role in determining whether, or how often, a particular plant is visited. For instance, the abundance and density of conspecific plants and the composition of neighbouring vegetation can also influence visitation, pollination and fruit set. The contribution of a plant to subsequent generations is determined by interactions with both mutualists (e.g. pollinators or seed dispersers) and antagonists (e.g. herbivores, florivores or seed predators), yet these associations are rarely studied in combination. Some traits or habitat characteristics that enhance a plant’s attractiveness or apparency to mutualists may simultaneously increase the risk of discovery by enemies.
This study simultaneously examines the influence of apparency on the pollination success and vertebrate herbivory of an endangered terrestrial orchid, Caladenia rigida R.S.Rogers. We assessed the effect of floral height, the local density of conspecifics, and concealment amongst neighbouring vegetation, on pollination and successful seed release among five orchid populations, over three years. Seed release was the net outcome of conflicting interactions with pollinators, florivores and fruit predators and varied both spatially and temporally.
However, we found no strong evidence for a compromise between pollination success and predation risk for any of the factors examined. Floral height had a positive effect on pollination success and the subsequent release of seed in one year, while both local density and concealment affected the risk of floral browsing in all three years of the study. Experimental removal of neighbouring vegetation did not affect the final proportion of plants that released seed, but increased the rate of discovery by florivores. Our data suggest that pollinators and herbivores respond to different visual and/or olfactory cues when locating flowers of C. rigida . Although pollinator-mediated selection for flower height is not directly disrupted by conflicting interactions with antagonists, selection pressures may be complicated or dampened by temporal variability and extrinsic characteristics of the plant population, or of the ecological community as a whole. Based on different predation rates for flowers and seed capsules, we propose that under intense grazing pressure, pollination soon after anthesis can provide plants with a reproductive advantage.
Diversity and evolution of floral venation in Apiales, with emphasis on Araliaceae
Nuraliev, M 1 , Oskolski, A 2 , Sokoloff, D 1
349

1 Moscow State University, Moscow, Russia; 2 Komarov
Botanical Institute, Russian Academy of Sciences, St
Petersburg, Russia
Although the pattern of floral vascular system is commonly considered now as an evolutionary labile feature of little importance for phylogenetics, a detailed comparative analysis of its diversity could be of great interest for understanding floral morphogenesis and its regulation, floral evolution and functional aspects of flower structure. We consider the order Apiales as a model group for analyzing evolution of floral vascular system. Both the published records and extensive original data on floral vasculature in different groups within Apiales (especially in Araliaceae) will be discussed in the context of molecular phylogenetic data.
Our analysis suggests that some (but not all) important transformations of floral conductive system are conditioned by the variations of flower groundplan (the number and position of sepals, petals, stamens and carpels). Both flower groundplan and vascular anatomy are uniform in Umbelliferae, the most prominent feature being presence of ten peripheral bundles in the inferior part of the ovary. Each of them ultimately divides to produce a branch to petal or stamen (and sepal, when present) and a carpellary branch. The entire floral venation in Umbelliferae (at least in Apioideae) should be considered in the framework of the occurrence of the specialized fruit type. Pentamerous tetracyclic flowers
(possibly except the dimerous gynoecium), similar to those in Umbelliferae are plesiomorphic in Araliaceae.
At least in four lineages of Araliaceae (Asian Schefflera clade, Plerandra group, Tetraplasandra , Osmoxylon ), polymerous flowers or flowers with polymerous androecia and/or gynoecia evolved (iteratively in some of the lineages) and other unusual transformations of floral structure took place. Vascular anatomy in representatives of each of four groups will be discussed. We found the following vascular features being variable at least in
Asian Schefflera species examined: presence of calyx bundles, number of petal bundles, origin of petal lateral bundles, presence of anastomoses between stamen and petal bundles, number of lateral carpel bundles, manner of ventral bundle fusion (homocarpellary or heterocarpellary), type of ovule innervation. In
Araliaceae, flower vasculature can differ considerably even between closely related species with isomerous pentamerous flowers. Most surprisingly, closely related
Schefflera species differ in types of petal venation that were previously considered to characterize major asterid clades. In several lineages of Apiales, the ovule bundles, which are primitively derived from the ventrals, tend to get their supply from lateral bundles of carpels.
Genetic models for the study of the reduction of petals and stamens in the evolution of Cruciferae
Penin, A 1
1 Dept of Genetics, Biological Faculty, Moscow State
University, Russia
There exists an immense diversity of flowers with respect to number and position of organs and their structure.
However, some families are characterized by a notable constancy of the flower. One of such families is
Cruciferae. The flower of most of its members consists of
350 four sepals, four petals, six stamens in two whorls (two short and four long) and a pistil consisting of two carpels.
However, in some species certain alterations are observed, for example in many representatives of genus
Lepidium a reduction of the short and some of the long stamens takes place. Such type of variation is characteristic not only of this family, but of the whole number of related families comprising the order
Capparales. It stands out that the observed alterations affect organs located in the intermediate zone of the flower – mostly short stamens, while organs located in the basal (sepals) or apical (carpels) parts always remain stable. We suggest studying the genetic basis of the reduction of petals and stamens using three models: 1)
Arabidopsis mutant lepidium-like (lel) that is characterized by the reduction of petals and part of stamens. 2) Naturally occurring variety of Capsella bursa-pastoris with reduced number of petals. 3) The species of the genus Lepidium characterized by different degrees of floral organ reduction. These three systems encompass all hypothetical stages of the speciation under the 'hopeful monsters' model – from single gene mutations that dramatically change plant morphology via populations to the species with the new morphological traits. The data on the morphology, inheritance and the expression of genes involved in floral development will be presented.
Mapping liverwort spore wall ultrastructure onto phylogeny
Taylor, W 1 , Strother, P 2
1 University of Wisconsin-Eau Claire, USA;
Observatory of Boston College, USA
2 Weston
Blackmore and Barnes in 1987 proposed that laminated sporoderm, corresponding to a type seen in mature extant liverworts, was the primitive condition in the embryophyte lineage. Laminated sporoderm has subsequently proven to be a common wall type in the early fossil plant record. It has been evoked by Wellman et al. in 2003 to assign liverwort affinity to Late
Ordovician cryptospore tetrads, and we have shown previously that laminated sporoderm is common in
Middle Cambrian cryptospore dyads. The possibility that variations in laminated sporoderm might be present in the fossil groups that were ancestral to the liverworts, inspired us to look more closely at this character as expressed in extant liverworts. We have gathered all available TEM images of spore walls from extant liverworts and mapped them onto the liverwort phylogeny of Forrest et al. 2006 as a means of determining the evolutionary history of spore wall construction in the liverworts. Initial results show that the
Riccia -type wall occurs in some Middle Cambrian cryptospore dyads. Given the liklihood that the Riccia type sporoderm is derived from a parallel laminate ancestral form, spore wall architecture would seem to support the liverwort–charophyte split before the Middle
Cambrian (Drumian Stage, ca. 505 Ma ±2 m.y.). These results support ongoing research indicating that the embryophytes evolved first from an amphibious gametophyte which later acquired a sporophytic phase through interpolation – a scenario proposed by Bower over a century ago.

Haptanthus hazlettii
, enigmatic Central American plant, in the light of new findings
Shipunov, A 1 , Oskolski, A 2
1 Minot State University, North Dakota, USA;
Botanical Institute, Russia
2 Komarov
Haptanthus hazlettii (Haptanthaceae) was collected in
1980 from Northern Honduras (Central America), and considered as a sole member of separate angiosperm family of uncertain position (Takhtajan, 2009). For twenty years, numerous attempts to recollect it failed.
Reproductive organs of Haptanthus are unique among angiosperms and make the search for relationships difficult. Unfortunately, all attempts to extract DNA and proteins from existing sample were unsuccessful. In
2010, Haptanthus has been finally re-collected, and preliminary molecular analysis of barcoding region of rbc L resulted in a robust placement of Haptanthus inside
Buxaceae, small family of basal eudicots. The strict position of Haptanthus within Buxaceae was not clarified. In MP analyses, Haptanthus form a common clade with all nonBuxus genera of this family
( Styloceras, Pachysandra and Sarcococca ) whereas in
ML analyses we obtained support for Haptanthus +
Buxus s.l. (incl. Notobuxus ) clade. The latter placement was confirmed by the data of wood anatomy and floral morphology: within Buxaceae, the South American genus Styloceras is the most similar to Haptanthus . In contrast to other members of family, these two genera share very long vessel elements (average length exceeds
1500 µm), heterogeneous rays of Kribs’ I type, the absence of a pistillode in male flowers, smooth transition between stamen filament and anther, massive sheath of endothecium and endothecium-like tissue surrounding each theca. It is noteworth that the flowers with two stamens occur also in male inflorescences of some
Styloceras species (von Baltazar & Endress 2002).
Comparison of Haptanthus and Styloceras male flower groundplans suggests that the paired anther-bearing organs in the former genus should be considered as flattened filaments rather than foliar structures with adnate stamens (Doust & Stevens 2005). However, this preliminary interpretation can be corrected after more detailed anatomical study of the Haptanthus flowers.
Testing methodologies for the estimation of undersampled geographical regions: the Asteraceae family in Australia
Schmidt-Lebuhn, A 1 , Gonzalez Orozco, C 1
1 CSIRO Plant Industry, Canberra, Australia
Inferring patterns of diversity, centres of endemism and the degree of sampling completeness for species rich plant groups is particularly challenging in large but unevenly populated countries like Australia. Sampling density is high near population centres, along roads and in areas close to research stations and herbaria, but drops sharply in more remote parts of the continent. To set priorities for additional sample collection, it would be useful to understand which areas have been underexplored relative to the expected species number. The
Asteraceae are the largest flowering plant family with an estimated 20,000–25,000 species worldwide, and they are represented in Australia with ca. 1,000 species from
200 genera. As the family is very diverse in arid areas, our knowledge of the distribution patterns of its species may be limited by patchy sampling activity. To assess the situation, we have extracted specimen data for all
Asteraceae collections from the online database
Australia’s Virtual Herbarium. Using conservative
(lumping) approaches to taxonomic circumscription, all specimens identified to species level and supplied with coordinates or sufficiently precise locality data are used to visualize species number registered for individual grid cells in Australia. In a second step, estimated species richness per cell is calculated using the Chao estimator.
Finally, the index of sampling completeness is calculated from observed and estimated species to identify underexplored areas. In a second approach, the Biodiverse tool is used to map spatial measures of biodiversity, and the results are compared to those from the previous analysis.
1
Predictive vegetation mapping
Cheal, D 1 , White, M 1
Arthur Rylah Institute for Environmental Research,
Melbourne, Australia
Mapping of vegetation community distribution has traditionally been done by extensive (and expensive) field work involving intense (personal and subjective) familiarity with the vegetation of a region. The vegetation communities thus defined and characterized, are then mapped, based on their remote visual signature, often using aerial fotografy, Landsat or similar techniques. The result is often the basis of land management, but is expensive, often highly subjective and restricted by the mapping scale chosen. We here present a novel method of vegetation mapping, as used for mapping the native vegetation occupying app. 970000 ha of East Gippsland in Australia. A long-term (dating back more than 20 years) database of floristic relevés was interrogated and over 4000 useful relevés extracted.
These were combined with around 100 novel relevés
(collected to provide geografic 'infill') and sorted and classified using phytosociological techniques. All relevés had a precise (geospatial) location. The locations of relevés from a phytosociological (vegetation) unit were then matched with a variety of geospatial datasets
(including rainfall, topography, radiometric data) and an environmental 'signature' for each vegetation unit identified. The resultant environmental signatures were then mapped and a vegetation map resulted. Subsequent ground truthing led to refinement of the units and improved maps. The advantages of this approach include
– much less expensive than traditional vegetation mapping techniques, – vegetation units can be readily recognized and mapped at different scales (e.g. 3 units vs. 30 units for the same region), – indicators obtained of the most significant environmental features that determine vegetation communities, – more objective maps (not reliant on individual interpretations of significant, usually visual, discontinuities).
351

When low copy nuclear genes are unable to produce species trees despite large genetic variation: lessons from a highly reticulate plant group (
Hieracium
)
Fehrer, J 1 , Krak, K, Chrtek, J
1 Czech Academy of Sciences, Czech Republic
A major challenge for the reconstruction of species relationships based on molecular phylogenies is presented by groups with recent speciation, abundant hybridization and the occurrence of polyploidy.
Hawkweeds ( Hieracium ) are among the most complex plant groups in these respects. Besides, the fixation of genotypes by apomixis has a major impact on the maintenance of alleles in taxa of hybrid origin. For the elucidation of species relationships and the origin of hybrid taxa, we used seven molecular markers: two chloroplast DNA regions ( trn T-L and trn Vndh C intergenic spacers), two nuclear ribosomal gene regions
(ITS and ETS), and three newly developed low copy nuclear (LCN) genes (glycine hydroxymethyltransferase
– shmt, squalene synthase – sqs, and gammaglutamylcysteine synthetase – gsh1). The three LCN genes provided the highest amount of variation; however, the trees were largely incongruent. The lowest variation was found with ITS and the two chloroplast markers. The best estimate of species relationships was provided by the
ETS tree. It revealed two major lineages with Eastern or
Western European origin and significantly different genome size which contained several subclades each and showed that many accessions had hybrid origin. The
LCN gene trees partly matched ETS subclades with taxa of hybrid origin clustering along with at least one parental species revealed by ETS. On the one hand, the
LCN genes divided the ETS subclades into two or more divergent lineages while on the other hand they merged taxa that belonged to different major ETS clades within a single lineage, with little variation among them.
However, the number of divergent lineages did not exceed the ploidy level of particular accessions. No evidence for pseudogenes or paralogs was found, and the divergent LCN variants were therefore interpreted as divergent alleles of heterozygous (or hybrid) individuals.
Relationships among these lineages remained unclear with all three LCN genes (no support for basal lineages, only for crown groups), i.e., neither was able to produce a species tree. According to LCN genes, the genus behaves like a single large population characterized by a rapid diversification of basal lineages and suffers from lineage sorting with a rather accidental distribution of particular alleles among the species. Interpretations are further complicated by the hybrid origin of many taxa. In addition, ETS, cpDNA and LCN genes all showed evidence for extinct diploid species whose ribotypes, haplotypes, or alleles were maintained in accessions with hybrid origin (including diploids), but were not found in any diploid non-hybrid species (only one diploid
Hieracium species was not sampled yet). While nuclear ribosomal DNA seemed to best reflect species relationships, the more variable LCN genes could provide deeper insight into the processes that have lead to the origin of particular species. In any case, much care has to be taken with the interpretation of LCN gene trees.
Each might provide a different aspect of speciation in a particular group, but to allow or at least facilitate interpretations, a reasonable species tree based on other
352 evidence (nuclear ribosomal DNA, morphology, geographic distribution etc.) is strongly recommended.
Morphology as an arbiter of gene tree incongruence: phylogenetic relationships and biome history in the
African flowering plant genus
Jamesbrittenia
(Scrophulariaceae: Limoselleae)
Verboom, GA 1 , Herron, ML 1 , Slingsby, JA 1
1 University of Cape Town, South Africa
The recognition that gene tree incongruence has a real, biological basis, rather than being a mere analytical or sampling artifact, casts doubt on the validity of analyzing multiple, contradictory gene loci in a single combined analysis ('total evidence'), especially where conflict is strong and spans deep nodes. Combining data in spite of conflict may compromise both topology and branch length estimation, with consequences for the quality of molecular age estimates. In this paper we employ a novel approach to resolving gene tree conflict which involves: identifying the set of taxa responsible for incongruence; splitting each of these into entities representing each locus; running a combined analysis to identify alternative placements of each ‘conflict taxon’ according to each locus; and identifying which placement, in each case, is most consistent with morphology. We justify the use of morphology as an arbiter of gene tree conflict on the grounds that it represents an integrated genotypic signal, based on multiple loci sampled from across the genome.
Application of the approach to Jamesbrittenia
(Scrophulariaceae), an African-endemic genus of flowering plants, shows morphology to be capable of resolving decisively the positions of several conflict taxa, even with a small number of characters. Comparison with other methods (concatenation, consensus and concordance) identifies our method as superior in terms of supported resolution, character consistency and taxon stability. Accuracy is difficult to assess, but is likely to be limited by morphological character quality and, as with other methods, taxon and gene tree sampling. A molecular dating analysis and reconstruction of historical biome shifts in Jamesbrittenia indicates a mid-Miocene origin in arid scrubby vegetation, probably along the margins of the Namib Desert. This supports an earlier interpretation of the Namib as a relatively ancient centre of origin for the southern African flora. The subsequent
(Miocene-Pliocene) radiation of Jamesbrittenia into a suite of vegetation types associated with the central
Escarpment (Grassland and Nama Karoo) and the southern margin of southern Africa (Albany Thicket and
Limestone Fynbos), may reflect the relative youth of these systems, their origins being tied to a major continental uplift during the Pliocene.
Systematic relevance of glandular trichome distribution, diversity and structure in
Cleome
(Cleomaceae)
Ivanova, A 1 , Koteyeva, N 1 , Shavarda, A 1
1
Voznesenskaya, E 1
Komarov Botanical Institute, Russia
,
Trichome morphology and distribution are reliable taxonomically important characters but structural

features of glandular trichomes seldom are considered as diagnostic traits. Overground parts of most Cleome species bear glandular pubescence. Distribution and morphology of trichomes of leaves, petioles and stems were surveyed in 48 Cleome species and among them in
12 species anatomy and ultrastructure of glandular trichomes were studied to evaluate their taxonomic potential. Four morphological types of glandular trichomes were recognized in Cleome : (1) small uniseriate thichomes, (2) middle-size biseriate thichomes,
(3) long-stalked trichomes with spherical head, (4) large trichomes with multicellular globose head. Distribution and structural characters of trichomes were compared to phylogenetic trees (Feodorova et al. 2010, in press). The following tendencies were revealed when the genus was surveyed from early- to late-diverging clades. In basal clade species ( C. africana and C. violaceae ), trichomes of type 2 were uniformly distributed over leaves, petioles and stems. Few ultrastructural features were related to secretion of secondary metabolites in head cells. No specific features of cytoplasm distinguished head from stalk cells. C. droserifolia belonging to early-diverging clade had trichomes of types 1, 2 and 3 but cell ultrastructure in trichomes of all three morphological types was similar to trichomes of other basal clade species. Species from North American clade ( C. isomeris, C. lutea and C. serrulata ) bore only nonglandular hairs on their leaves and stems. In the sister clade, C. angustifolia and C. paradoxa had glabrous leaves but singular trichomes of type 4 were found on their stems. Despite morphological difference in maturity of these trichomes from other trichome types they developed as biseriate glands in the early stages of their ontogenesis. C. viscosa , C. gynandra and C. foliosa had uniformly distributed glands of morphological type 2.
Cell specialization appeared in their heads: the upper cells acquired common features of terpenoid-secreting cells unlike cells of the lower part of the head. Leaves of
C. hassleriana from the late-diverging clade bore type 2 trichomes on the leaf blade and type 3 at leaf base and petiole. Ultrastructure of their head cells differed dramatically: it was typical to terpenoid-secreting cells in the latter and it was similar to trichomes from basel clades in the former. There were dilated reticular cisternae resembling myrosin bodies of Brassicaceae in gland head cells of some species; substances that supposed to be glucosinolates or their derivates have been found in methanolic extracts of leaves by GC-MS.
Trichome distribution and morphology as well as anatomy and ultrastructure of glandular trichomes are evident diagnostic traits for Cleome . They agree with the existing taxonomy of the genus (Iltis 1952) and support new phylogenetic data (Feodorova et al. 2010, in press).
The main structural characters of glandular trichomes are conservative within the genus. Though ultrastructure of secretory cells reflects composition of secretion and therefore is determined by ecology of the species, the phylogenetic patterns are obvious on ultrastructural level.
The main trend of trichome evolution in Cleome is specialization of morphological types in relation to the composition of their secretion.
Genetic reconstruction of colonization patterns of the
Neotropical epiphytic orchid,
Brassavola nodosa
Trapnell, D 1 , Hamrick, JL 1
1 University of Georgia, USA
Anthropogenic habitat disturbance is a common feature of many landscapes today and is responsible for putting numerous species at risk. The abandonment of an anthropogenically disturbed landscape creates a valuable opportunity to study patterns of natural colonization. As the original colonizing species arrive and establish, they alter the landscape in a variety of ways, including providing suitable habitat for subsequent colonists.
Epiphytic plants, which are major contributors to overall biodiversity in the Neotropics both directly and indirectly, are excellent exemplars of this phenomenon.
Only when suitable substrate (i.e. a host tree) is available can epiphytes colonize the landscape. Orchids are one of the major constituents of this epiphyte flora. As cleared areas, such as pastures, are re-colonized by suitable host trees, the establishment of epiphytic orchid populations, and their genetic composition, will be a function of seed dispersal and colonization patterns, in particular the number of founding individuals. Understanding factors that effect and limit colonization by epiphytic orchids may have important conservation implications. To address these questions we used genetic analyses to discern the prevailing pattern of colonization of suitable habitats by the neotropical epiphytic orchid Brassavola nodosa . Leaf samples (mean = 41.5 per population; range
= 9–56) were collected from three populations (i.e. trees) from each of four pastures in the dry forest of northwestern Costa Rica. All 12 host trees belong to the same species ( Crescentia alata , Bignoniaceae) and were separated by a maximum distance of 7.23 km. Leaf samples were assayed for their multilocus allozyme genotypes using one monomorphic and 18 polymorphic loci. Spatial autocorrelation analyses were used to assess the relatedness of individuals within populations compared with individuals from other trees and pastures.
The results showed significant levels of relatedness
(mean r = 0.168; range = 0.106–0.248) within host tree populations and significant but much lower values (mean r = 0.071; range = 0.011–0.140) between populations in the same pasture. Our data suggest that the colonization of new populations is by a few individuals (1–4 per tree) with subsequent in situ population expansion and that colonists within a tree were not a random sample of the regional seed pool. Furthermore, populations within the same pasture were either colonized by propagules from the same source or by seeds produced by the founders of a neighboring population within that pasture. These results have important ramifications for understanding the conservation measures needed for this species as well as other epiphytic orchid taxa.
Disentangling demographic processes and local adaptation in the evolution of adaptive traits in the model species
Arabidopsis halleri
Decombeix, I 1 , Pauwels, M 1
Saumitou-Laprade, P 1
, Godé, C
, Frérot, H 1
1 , Bourceaux, A 1 ,
353

1 GEPV Lille, France
In the Brassicaceae family, A. halleri is of particular interest because of its ecological, phenotypic, and genetic characteristics. In particular, the species is a
'pseudometallophyte': it grows on non-polluted areas as well as on calamine soils with high concentrations in zinc
(Zn), lead (Pb) and cadmium (Cd). Because these Trace
Metal Element (TME), either essential (Zn) or not (Pb,
Cd), are all toxic at concentrations encountered on calamine soils, the species is called 'metal tolerant'. A. halleri also displays metal hyperaccumulation capacities, i.e., it can absorb, translocate and accumulate high amounts of Zn and Cd in its shoot. Metal tolerance and hyperaccumulation are commonly considered as adaptive traits. Because of its phylogenetic proximity with A. thaliana , A. halleri is considered as a model species to study genetic bases of adaptation to calamine soils. In controlled conditions, all populations tested so far show metal tolerance and hyperaccumulation abilities.
However, mean quantitative differences have been noticed between metallicolous (M) populations growing on calamine soils and non-metallicolous (NM) populations, suggesting local adaptation. In this study, we analyzed the spatial distribution of quantitative polymorphisms for tolerance and hyperaccumulation traits among 25 M and NM populations at a local scale, in order to detect a signal of local adaptation. First we have conducted experiments in controlled conditions to determine zinc tolerance and hyperaccumulation polymorphism within and between the same populations.
Apart from metal concentrations, polluted areas can also show extreme values at many environmental parameters, like soil granulometry (polluted areas are often stony and thus very dry). We thus scored a large number of biotic and abiotic parameters in order to describe and to characterize sampling locations and to identify parameters which could discriminate population groups.
The population genetic structure was also analyzed using neutral markers in order to disentangle the respective roles of demographic history and local adaptation in the distribution of genetic polymorphisms. Nineteen microsatellites markers were used to genotype twenty individuals per population. Our results showed that (1) important phenotypic variations were evidenced with a structured polymorphism according to both geographic distance and edaphic type, (2) although soil elements are major discriminating factors, A. halleri habitat is diverse,
(3) at a local scale, neutral genetic structure is mainly determine by geographic distances; populations on calamine and non-calamine soils are thus highly connected. However, maybe because of a high heterogeneity between and within populations and/or a lack of statistical power, we could not be able to detect a clear signal of local adaptation in our studied populations. Moreover, our results show that TME are not the only selective pressure, and that other criteria should be taken into account in the ecological differentiation between 'metallicolous' and 'nonmetallicolous' populations of A. halleri .
Determining the origin and diversification of
Murraya paniculata
: one or more species?
Nguyen, CH 1
Mabberley, DJ 2
, Beattie, GAC 1
, Weston, PH 3
, Holford, P 1 , Haigh, AM 1 ,
354
1 Centre for Plants and The Environment, University of
Western Sydney, Australia;
Kew, UK; 3
2 Royal Botanic Gardens,
National Herbarium of New South Wales,
Sydney, Australia
Murraya paniculata (L.) Jack [Rutaceae: Aurantioideae:
Aurantieae] occurs naturally in Asia and Australasia. It is also a widely cultivated ornamental plant known by the common names orange jasmine, orange jessamine and mock orange. The ornamental form is the preferred host of the Asiatic citrus psyllid ( Diaphorina citri Kuwayama
[Hemiptera: Psyllidae]), which in Asia and the Americas is the major vector of ' Candidatus Liberibacter spp.' [
α
-
Proteobacteria] pathogens that cause huanglongbing
(HLB), a devastating disease of citrus. Some authorities regard the ornamental form as a distinct species, M. exotica L. Recent studies indicate that this ornamental form is a transient host of liberibacters whilst other studies suggest that M. paniculata is not a host of these pathogens. A subspecies, M. paniculata subsp. ovatifoliolata Engl., is considered indigenous to Australia and its host status with regard to the liberibacters is unknown. Other subspecies of M. paniculata occur in
Asia and their susceptibility is also unknown. Due to the severity of HLB and its consequences on cultivated citrus and more broadly on the biodiversity of Rutaceae, this has led us to review the status of taxa currently regarded as being M. paniculata . Diversification within M. paniculata (sensu lato) may be related to tolerance or susceptibility to the liberibacters and knowledge of this may lead to novel forms of pathogen control. To understand the diversification of M. paniculata , our research focused on the phylogenetic relationships among accessions of specimens collected within
Australia and overseas. Six regions ( trn Trps 4R2, trn CGCAycf 6R) and spacers ( trn L-F, rps 16, ma tK-
5' trn K, psb M -trn DGUC) of the maternally-inherited chloroplast genome have been amplified by PCR from a total of 86 accessions and the resulting amplicons sequenced. In addition, part of the internal transcribed spacer (ITS) region of the nuclear-encoded ribosomal
RNA operon has also been sequenced. The data obtained were subjected to parsimony and Bayesian analysis to determine phylogenetic relationships; these two methods produced cladograms with similar topologies. Results from the chloroplast genome indicate that there are two distinct clades, Paniculata and Exotica. Within the
Paniculata clade, three subclades were found: a subclade containing all accessions from Indonesia identified as M. paniculata ; a subclade containing the large and small leaf forms of M. paniculata subsp. ovatifoliolata from
Australia, M. paniculata subsp. omphalocarpa from
Taiwan and M. paniculata subsp. zollingeri from
Indonesia; a subclade containing accessions from Cuc
Phuong, Vietnam, Yingde China and Pakistan. The phylogenetic analysis based on ITS sequence data also places the accessions into two major clades, membership of which is similar to that of the Paniculata and Exotica clades from the cpDNA analyses. However, in the ITS analysis, a hairy accession of M. paniculata from
Indonesia that falls in the Exotica clade according to chloroplastal sequence data now appears in the
Paniculata clade. Conversely, the subspecies omphalocarpa falls with the Exotica clade. These anomalies may be due to hybridisation events or to incomplete lineage sorting. We conclude that that M.

paniculata and M. exotica are distinct species. However, this needs to be tested through further phylogenetic analyses together with morphological and phytochemical studies.
Evolutionary history of
Haplophyllum
(Rutaceae,
Sapindales) in the Irano-Turanian and
Mediterranean floristic regions
Manafzadeh, S 1 , Salvo, G 1 , Conti, E 1
1 Institute of Systematic Botany, University of Zurich,
Switzerland
Sapindales, one of the largest angiosperm orders (ca. 3% of eudicot diversity), include prominent elements of the tropical floras of the world, mostly woody plants.
Rutaceae is the largest family in the order (161 genera and 1815 species), and is also mostly tropical and subtropical in distribution. Haplophyllum is one of the most species-rich, but least-studied genera of Rutaceae.
Its habit (herbaceous) and habitat (arid regions of the
Northern Hemisphere) differ from the more typical members of its family and order. It includes 68 species distributed throughout temperate and subtropical Eurasia and few tropical species of north-eastern Africa. It reaches maximum species diversity in the Irano-Turanian floristic region – in particular, Iran, Turkey, and Central
Asia – which harbour 60% of the species diversity.
Twenty one percent of the species are present in the
Mediterranean basin. Many species of Haplophyllum are characterized by a small geographical range size, sometimes limited to a single mountain chain (i.e.
‘narrow endemics’), a feature that makes them especially vulnerable to extinction, underscoring that, if left understudied and unprotected, they are under threat of extinction. We are performing an integrated study that will allow us to understand the origin and evolution of
Haplophyllum , define species boundaries, improve understanding of species diversity, and finally recommend conservation guidelines. Consequently, we generated phylogenies from DNA sequences of four regions of the chloroplast genome for 66% of the species of the genus (45 species, 118 accessions). To improve our understanding of taxonomic species boundaries,
Haplophyllum was studied morphologically: a matrix was constructed for 27 discrete morphological characters scored for the same species used in the phylogenetic analyses. Moreover, molecular dating, and ancestral range reconstructions analyses were carried out to investigate the extent to which past geological and climatic histories of the Irano-Turanian and
Mediterranean floristic regions explain the current distribution of the genus. Our phylogenetic analyses identified both cases of strongly-supported species monophyly and instances of species non-monophyly.
Character mapping analyses indicated that the main morphological characters traditionally used to classify the genus are consistent with the molecular phylogeny of
Haplophyllum . The Mediterranean representatives of
Haplophyllum did not cluster together and were found to be embedded within a clade that includes primarily
Irano-Turanian species. This suggests that multiple invasions of the Mediterranean basin from the east took place during the evolution of the genus. Our preliminary results strongly suggest that the genus Haplophyllum originated in the Irano-Turanian region and subsequently colonized the Mediterranean basin. This is the first study on the interaction between two floristic regions via a genus that has originated in one region and colonized the other. Different ecological requirements (desert vs. mountainous species) and diverse geographical ranges
(widespread vs. narrow endemic) make Haplophyllum an excellent model to investigate the correlates of habitat range with the geography of speciation (sympatric, allopatric, or peripatric modes).
Phylogeographic patterns of native banana species
(
Musa
L., Musaceae) in Thailand
Swangpol, S 1 ,
Rotchanapreeda, T
Boonprasert, R
1
2
, Atawongsa, K 1
, Yanisarapunt, S 1
, Wongniam, S 1
,
1 Dept of Plant Science, Mahidol University, Bangkok,
Thailand; 2 Faculty of Environment and Resource
Studies, Mahidol University Salaya Campus, Nakhon
Pathom, Thailand; 3 Dept of Botany, Chulalongkorn
University, Bangkok, Thailand; 4 Dept of Biochemistry,
Mahidol University, Bangkok, Thailand
Thailand, situated at the heart of the mainland Southeast
Asia, is among the origins of wild bananas ( Musa L.,
Musaceae) where at least seven species and three subspecies occupy habitats mostly in mountain ranges lying in north-to-south direction. To determine specific ecology and elucidate geographic distribution patterns in association with gene genealogy of the bananas, morphometric and molecular analyses were employed.
Biogeographically, two or more species or subspecies have been found growing in sympatry in many localities among over 100 sites investigated. The analyses indicated that phylogeographic patterns emerged in M. acuminata Colla with subsp. siamea Simmonds found in northern to central Thailand while subsp. malaccensis
(Ridl.) Simmonds typically inhabiting lower peninsula
Thailand. Unexpectedly, some accessions of subsp. malaccensis were also found in northern border of
Thailand–Myanmar. The incident evoked further discussion on the dispersal of the subspecies through the bordered mountain chain. In addition, within this subsp. malaccensis , a new genetically distinct form was found in the Kra Isthmus, at the boundary of Indochinese and
Sundaic floristic provinces. Moreover, color-deficient mutants, two new records, and possibly a new banana species were reported here. Accordingly, preservation of the banana 'hotspots' will definitely guarantee the existence of species diversity at the cradle of its origin, and further utilization for many breeding programs may render.
Affinities of the flora of the Guiana Shield, northern
South America: Boraginaceae, Gesneriaceae &
1
Passifloraceae
Feuillet, C 1 , MacDougal, JM 1 , Skog, LE 1
Smithsonian Institution, Washington DC, USA
The Guiana Shield is a very old geological formation made of crystalline rocks later covered by thick sandstone of marine origin that covers the north of
Brazilian Amazonia, the Venezuelan Guayana and the three Guianas. Later the sandstone layer was partially eroded, leaving tepuis (steep-sided table mountains) in
355

the west. Although the region is surrounded with areas with similar climate and vegetations, the endemic part of the flora is important. The vascular plants are 40% endemic.The Boraginaceae, unplaced in the Lamiids, present strong variations in the level of endemism between the subfamilies: Heliotropioideae 0%,
Cordioideae 49%, Ehretioideae 100%. The Gesneriaceae, in the Lamiales (Lamiids), include 60% of endemic species, and the Passifloraceae–Passifloroideae, in the
Malpighiales (Fabids), 43%. In the same way, the distribution of the species present in a larger area have different affinities in the 5 groups.
Evolutionary history of the Hawaiian violets
Havran, J 1 , Ballard, H 2 , Sytsma, K 3 , Sack, L 4
1 Campbell University, North Carolina, USA;
University, USA; 3 University of Wisconsin–Madison,
USA; 4 University of California at Los Angeles, USA
2 Ohio
The Hawaiian violets ( Viola , Violaceae) are a monophyletic group of nine taxa that have undergone adaptive radiation across the six high islands of the
Hawaiian archipelago. Previous research revealed convergences in leaf and stem morphology among different taxa in similar habitats. We conducted a multidisciplinary study involving all known species of
Hawaiian violets to describe the biogeographic patterns of leaf form and functional trait evolution in the context of an adaptive radiation. We elucidated the historical biogeography of the Hawaiian violets through a molecular systematic investigation based on the Internal
Transcribed Spacer sequence of violet populations from each Hawaiian Island. We determined a Maui Nui origin for the Hawaiian violets, with subsequent dispersals to
Kauai, Oahu, and Hawaii, using Maximum Likelihood and Bayesian inference reconstructions. Establishment on
Maui Nui would indicate a maximum age of the
Hawaiian violets of 1.2 million years. The monophyletic group is separated into two subclades (a wet clade and a dry clade) distinguished primarily by differences in precipitation regimes. Within the wet clade, representing species distributed in high-elevation bogs, cloud forests, and mesic streambanks, Bayesian inference reconstructions indicated the basal leaf shape morphology was either petiolate ovate or petiolate lanceolate. The evolution of petiolate orbicular and petiolate ovate leaf morphologies is associated with habitat shifts to bog and cloud forest habitats, respectively. These habitat shifts have occurred twice within the Hawaiian violets: one on Kauai and another on
Maui Nui. We also measured more than 50 leaf functional traits for eight Hawaiian violet species from bog, cloud forest, dry forest, and cliff habitats. We found a strong diversification among species in all leaf traits measured, even among sister species pairs within the wet clade and the dry clade. Species in the wet clade exhibited thinner cell walls and thicker leaves than those species in the dry clade. The strong coordination between leaf traits in the wet versus the dry clade indicates clusters of co-evolving traits among species with narrow ranges of soil moisture and precipitation. In parallel with convergences in leaf form, we found a convergence of physiological traits, including leaf hydraulic conductance and light saturated photosynthetic rate per area, between bog ( Viola kauaensis and V. maviensis ) and cloud forest
( V. wailenalenae and V. robusta ) taxa on Kauai and Maui
Nui. When compared with cloud forest taxa, violets adapted to bog habitats exhibited smaller leaves, higher leaf hydraulic conductance, and higher gas exchange capacity than cloud forest species. Leaf shape and functional traits that have been repeatedly derived in bog taxa may be associated with high water availability and high irradiance in high-elevation open bogs. The relatively young age of the Hawaiian violets (1.2 million years) derived from molecular systematic investigations indicates a relatively rapid adaptive diversification of leaf features that included convergent evolution of morphology and physiology in open bogs and cloud forests across the Hawaiian archipelago.
Phylogeny and character evolution in
Thysananthus
1
(Lejeuneaceae) – back to the future with modern data and the first hybrid in the largest liverwort family
Sukkharak, P 1 , Gradstein, SR 2 , Stech, M 3
Albrecht von Haller Institute of Plant Science,
University of Goettingen, Göttingen, Germany; 2 Museum
National d'Histoire Naturelle, Dept Systematique et
Evolution, Paris, France; 3 Netherlands Centre for
Biodiversity Naturalis, section NHN, Leiden University,
The Netherlands
The first comprehensive molecular phylogeny of
Thysananthus Lindenb. (Marchantiophyta: Lejeuneaceae subfamily Ptychanthoideae) is assessed by sequence analyses of five markers ( psb Atrn H spacer, trn G intron, trn L-F, trn Srps 4, nrITS) and 27 accessions from all ten currently recognised species, including the recently described T. discretus Sukkharak et Gradst. The species of Thysananthus are resolved into two major clades, which also include Dendrolejeunea fruticosa and
Mastigolejeunea pancheri . The clade of D. fruticosa, T. mollis, T. montanus and T. retusus stands out by the presence of vitta and corresponds to a section recognised in early classifications. The well-supported sister relationship between the T. anguiformis – M. pancheri and
Thysananthus s. str. clades is noteworthy as M. pancheri has long been separated from Thysananthus by its entire female involucres and presence of a stem hyalodermis.
One specimen of M. pancheri is probably of hybrid origin, which would be the first report of a hybrid in the largest liverwort family. Within Thysananthus s. str., T. comosus, T. convolutus and T. spathulistipus were polyphyletic, but the molecular clades largely fit with formerly recognised (but later synonymised) species, and in one case with a novel species that is also recognisable morphologically. In the sister genus Mastigolejeunea , accessions of M. auriculata from southern USA and
Malaysia are not in the same clade, indicating that the
New World populations differ genetically from the Asian ones that were originally described as M. humilis . The results of ancestral state reconstructions suggest that a number of morphological characters currently used to distinguish genera in Ptychanthoideae are homoplastic.
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Phylogenetic diversity of the ectomycorrhizal fungus
Cortinarius associated with native and introduced forests in New Zealand
Orlovich, D 1 , Teasdale, S 1 , Lucas, M 1
1 University of Otago, USA
Soil microbiota mediate interactions between plants and soil, and are thus essential for native ecosystems and managed agriculture. Many forest trees require ectomycorrhizal fungi for growth and survival, yet there is limited understanding of the relationship between fungal diversity and forest type. Ectomycorrhizal fungi are essential for the survival and vitality of native (e.g.,
Nothofagus ) and introduced ( Pinus radiata ) forests, whereas other forest trees (e.g., Leptospermum and
Kunzea in New Zealand) can live with or without ectomycorrhizas. We are discovering how phylogenetic diversity of ectomycorrhizal fungi varies between forest tree species, and what implications this has for understanding the dynamics of forest tree movement
(e.g., the spread of Nothofagus into Leptospermum forest). We have designed genus-specific primers to amplify key ectomycorrhizal genera, and have compared the phylogenetic diversity of Cortinarius (the most common and diverse fungal genus associated with
Nothofagus ) between forests containing Nothofagus menziesii / N. solandri, Kunzea ericoides , and Pinus radiata . We found surprisingly high degree of overlap between fungi occuring in all three forest types, a result not expected on the basis of taxonomic (i.e., formally described species with known host) diversity. We found that Kunzea ericoides was host to a greater-than-expected array of Cortinarius species, indicating that the taxonomic diversity of fungi associated with this host tree is underappreciated. We also found a difference between fungal diversity found by analysis of ectomycorrhizal Nothofagus root tips versus that found by cloning from soil-extracted DNA. Our results so far indicate that taxonomic diversity is a poor metric for comparison of fungal diversity between host tree types.
Studies on the biogeography of Nothofagus give little attention to the diverse communities of ectomycorrhizal fungi assocated with them, which is surprising since the tree is obligately ectomycorrhizal in the natural environment. Through our research we hope to refine tools for exploring and discovering ectomycorrhizal diversity, to better understand the role these fungi play in forest ecosystems, and to determine how fungal distribution patterns inform our understanding of forest biogeography.
Phylogeny of
Nemacladus
(Nemacladoideae,
Campanulaceae)
Morin, N 1 , Ayers, T 2
1 Flora of North America Association, USA; 2 Dept of
Biology, Northern Arizona University, Flagstaff,
Arizona, USA
Much of the diversity found in one or another of the subfamilies of Campanulaceae are found in
Nemacladoideae: Flowers resupinate or not (only
Lobelioideae are resupinate), with highly zygomorphic to nearly actinomorphic corollas (Lobelioideae,
Cyphioideae and Cyphocarpoideae are zygomorphic,
Campanuloideae are actinomorphic), glabrous style (all except Campanulaceae) that curves at apex and elongates after anthers dehisce (Lobelioideae), soft hairs on stamens (Cyphioideae), pollen presented on unopened stigma, superior to inferior ovary. Nemacladoideae also has features unique in the family: filaments partially fused (a few Cyphia basally connate) and anthers free and reflexed, large, clear cells attached to filaments by an elaborate tissue structure, large glands at the base of the style. Nemacladoideae comprises the annual Nemacladus
(ca. 17 species, including Parishella ), endemic to southwestern North America, and monotypic perennial
Pseudonemacladus of central Mexico. It is one of the three subfamilies of Campanulaceae that are phylogenetically and geographically isolated, together with the monogeneric African perennial Cyphioideae and the annual monogeneric Chilean Cyphocarpoideae.
Nemacladus occurs in deserts, mixed conifer forests, subalpine rock scree, and chaparral, 0–2700 m elevation, in southwestern U.S. and northern Mexico. The center of diversity is southernmost California and northern Baja
California. Only a few species are abundant even in years of ample rain. We wanted to reconstruct the phylogeny to test whether morphology and geography accurately reflected evolution. Were species with small flowers closely related? Were the species with highly zygomorphic flowers related? Were all the desert species related? At least one population of all 21 described taxa and several new taxa were collected; SEM and high resolution digital images of flowers were made; and about 2800 individual herbarium collections were examined. Phylogenetic analysis was done using nrITS and the cpDNA spacer atpB-rbcL. Two major clades were recovered: one primarily distributed in northern
California and higher elevations, with one species widespread in deserts, had one subclade with resupinate flowers and one with non-resupinate flowers and species pairs each with one large and one small-flowered member. Parishella californica nested in the nonresupinate subclade. The other major clade occurs in southwestern deserts and has species pairs each with a resupinate, usually smaller unmarked flower and a nonresupinate, larger, highly marked flower. The basal clade within this desert group consists of N. longiflorus with superior ovary, zygomorphic long-tubed corolla, and pink markings, and N. pinnatifidus with partially inferior ovary, nearly actinomorphic, short-tubed and unmarked corolla, both species with resupinate flowers. In general, non-resupinate flowers are mostly highly zygomorphic, have deeply cut, highly marked corollas, and straight filament tube and style and could be expected to be outcrossed whereas resupinate flowers have overarching filament tube and style and unmarked nearly actinomorphic corollas and could be expected to be selfing. A shift from non-resupinate, zygomorphic flowers to resupinate, nearly actinomorphic flowers could drive a change in pollinators, leading to rapid speciation. Such combinations of elements of flower morphology found elsewhere in the family, together with unique features, have resulted in the remarkable diversity found in Nemacladus .
Pollen morphology for systematics in the family
Violaceae
Gavrilova, Olga 1
357

1 Komarov Botanical Institute, Russia
Pollen of 75 species of the family Violaceae have been investigated by light, scanning and partly transmission electronic microscopy. Studied species belong to 10 genera from Leonioideae and 2 tribes of Violoideae subfamilies, most of the studied species are from the genus Viola . Electron microscope data add substructural evidence to palynological and taxonomical diagnostic.
We have found the following pollen morphological types: (1) 3-colpate with microperforate and/or microechinate ornamentation (neotropical Rinorea species, partly species of section Viola of genus Viola ).
(2) 3-(4)-colporate with microperforate and/or microechinate ornamentation ( Agatea, Hybanthus,
Leonia , Melicytus, Noisettia, Schweiggeria , Asian,
African and a part of neotropical Rinorea species, section
Dischidium and a part of section Viola of genus Viola ).
(3) 4-5-(6) colporate with microperforate or rugulate ornamentation (section Melanium of genus Viola ). (4) 2porate with microperforate ornamentation ( Amphirrhox and Paypayrola species). Types 1 and 2 are close to one another by form, sizes, apertures number, ornamentation and exine structure, but type 2 is more advanced by occurrence of compound apertures. Type 3 differs in all thses characteristics, including kinds of microperforate ornamentation, apertures and sporoderm structures. Type
4 is rare in dicotyledonous pollen. So, in Violaceae we have found two groups palynologically independent from other family members: section Melanium of genus Viola and subtribe Paypayrolinnae (genera Amphirrhox and
Paypayrola ).
Origin and evolution of
Cyananthus
(Campanulaceae), an endemic genus from Sino-
Himalayas
Sun, H 1 , Zhou, Z 1 , Niu, Y 1 , Wen, J 2 , Hong, D 3
1 Key Laboratory of Biodiversity and Biogeography,
Kunming Institute of Botany, Chinese Academy of
2 Sciences, China; Dept of Botany, National Museum of
National History, Smithsonian Institute, Washington DC,
USA; 3 State Key Laboratory of Systematic and
Evolutionary Botany, Institute of Botany, CAS, Beijing,
China
Cyananthus (Campanulaceae) is a genus strictly limited to the Sino–Himalayas, distribution range from SW
China extending westwards to Kashmir along the
Himalayas. It is the sole genus with a superior ovary in
Campanulaceae, so it is assumed to be the most primitive genus of the family. The genus consists of 19 species by latest morphological revision and is divided into three sections based on the life history, corolla lobes and hairy types on calyx: (1) Sect. Cyananthus , perennial herbs, corolla lobes usually oblong, longer than broad, calyx glabrous or covered with hairs other than brown-black setae, distributed from SW China to Himalayas ; (2) Sect.
Stenolobi , perennial herbs, corolla lobes nearly orbicular, broadly ovate or oblong, as long as or slightly longer than broad, calyx densely covered only with brown-black setae, only distributed in Eastern Himalayas; (3) Sect.
Annui , annual herbs, distributed mainly in Hengduan
Mountains, SW China. The nuclear ITS and four plastid markers ( mat K, rbc L, psb Atrn H, trn G-S) fragments were analyzed with parsimony, likelihood, Bayesian
358 inference to test the monophyly and evaluate the relationships within the genus. Three plastid markers
( mat K, rbc L, atp B) were conducted with parsimony and relaxed Bayesian dating to assess the systematic position of Cyananthus within the family and conduct the biogeographic diversification history of the genus. Our results suggest the genus is obviously monophyletic and the boundary of each section is also very clearly displayed in our systematical tree based on three chloroplast genes. Sect. Cyananthus and Sect. Annui form a clade, and Sect. Stenolobi forms a clade itself.
The monophyletic of the genus and each section has a high support value. The genus is closely related to
Codonopsis, Platycodon, Leptocodon whose ovary have more or less degrees of superioriority. All these genera are relatively primitive in the family. Our dating analyses demonstrate Sect. Stenolobi diverged from the other two sections in the late Miocene (about 7.27 mya) and Sect.
Annui diverged from Sect. Cyananthus in early Pliocene
(about 4.25mya). We conclude Sect. Stenolobi may be the primitive type of the genus which originated in the
Miocene (about 11.71 mya), Himalayas might be the earlier divergence place of the genus, then migrating to
Hengduan Mountains, evolving into two life history styles, Sect. Cyananthus with perennials and Sect. Annui with annuals. The latter two sections are still in fierce diversifications, especially in Sect. Cyananthus , low level of divergence in both morphology and molecular is shown in this young section. We further found that many
(and only) species belongs to Sect. Cyananthus include two sexes within population, hermaphrodites and females, and is therefore gynodioecious system. This sexual system has been regarded as an important link in transition from hermaphrodite to dioecy. The divergence in sexual system may lend additional thread to understand the evolution of this genus.
The plants of Mexico, as seen through the eyes of the artist
Silva-Torres, B 1
1 Universidad Autonoma Metropolitana, Mexico
Mexico is one of the most bio-diverse countries of the world, in fact occupying fourth place in terms of plant diversity. This diversity springs from the confluence of two biogeographic zones: the Neartic and Neotropical zones which result in flora ranging from desertic to tropical. This rich diversity has over the centuries inspired artist and in turn enriched México’s cultural history. Some painters have expressed their fascination with the natural world through the all-encompassing art of landscape. Two of the greatest masters of the Mexican landscape are José María Velasco and Gerardo Murillo
(generally know as Dr Atl). The variety and sheer number of Mexican artists who have studied the landscape mirrors the bio-diversity of México itself. Paul

Fischer, José Miguel Covarrubias and Rufino Tamayo may be mentioned. Some artists such as Diego Rivera have used nature to express purely symbolic meaning.
This may further a rhetorical, even a political program – a common impulse in the art of early and mid-century
Mexican art. But many artist have used natural motifs in a celebratory sense, free of symbolic import. Rufino
Tamayo painted succulent watermelons; Frida Khalo incorporated bouquets and fruits into her personal dialog.
Maria Izquierdo’s still life paintings reflect an appreciation of the life-giving nature of the flora of
México. Keith Miller and Leonard Brooks amongst other expatriate artists living and working in México have explored the incredible variety and strangeness of this country’s diversity. In my presentation I would like to suggest to the attendees the tremendous natural richness of México as seen through the eyes of her artists.
The legend of the sad tree in the 16th century
‘discovery’ of India by Europeans
Dias, AS 1
1 Universidade de Évora, Portugal
A peculiar tree with delicate white flowers of wonderful fragrance spreads its perfume at the Indian nights. The flowers open only at night and moreover on daytime the tree seems wilted as if the sunlight harms it. Given these characteristics, the Portuguese called it ‘arvore-triste’ which means sad tree and in the same sense Linnaeus gave it the name, still in use, of Nyctanthes arbor-tristis .
In his book Coloquios dos simples, e drogas he cousas mediçinais da India… on the medicines of India, first published in Goa in 1563, the Portuguese physician
Garcia da Orta (garçia dorta) devotes a chapter to the sad tree also known as tree-of-sorrow and night-Jasmine where he presented a legend told in Goa about this plant, that he considers as nonsense of those people. This people, he wrote, believes that this tree was a maiden, daughter of a noble man called Parizatak, who committed suicide after being snubbed by her lover, the sun, who dumped her for another. And from the ashes this tree was born and its flowers hate the sun and they do not appear in daylight. The notice of the tree and its legend was translated and commented by various European authors denoting the interest in an exotic Indian plant by a 16th century Europe, eager for the prodigious curiosities of the recently ‘discovered’ distant lands. Some of these authors also did illustrations of the plant where the botanical interest prevails but also happens that the illustration departs from the botanical realism and clearly attempts to represent the legend. The strong elements of the myth are discussed considering the European reaction to what can be seen as a metaphoric explanation for the set of peculiar features of this remarkable and useful plant. Another and far more complex Indian myth can be related with this tree that is considered as the Parijata tree that according to the Hindu mythology perfumes the entire universe. However, the Goa legend told by Orta, on its apparent simplicity has a set of features that makes it a human universal history worthy of all the attention dedicated to it. Namely the existence of a relationship with a superhuman entity, the sun, a love to death story, a death that gives place to rebirth, the spirit that prevails over death and rebirth, and finally a special tree that appears as the bright result of suffering transcendence, a world wonder and a gift to mankind.
Interpretation of the relationships between, plants, mythology and art
Planchuelo, A 1
1 CREAN, Universidad Nacional de Córdoba, Argentina
The relationships between Botany and Greek and Roman mythology are numerous and can be classified into the following categories: (1) Mentions of plants in mythological tales; (2) Mythological Gods or Goddesses associated with nature and plants; (3) Botanical terms related to mythology; (4) Species or genera names given by botanists in honor to mythological characters; (5)
Metamorphosis of mythological characters into plants.
The Renaissance and Mannerism (1400–1610) as well as
Neoclassical (1830–1890), art periods have many pictorial testimonies and sculptures of Greek and Roman mythological characters related to plants. Art recreation of original tales sometimes were adjusted to modern life and consequently confusion and misleading of the plants involved in the original Greek or Latin documents are common. Several plants are cited in mythological tales, like oaks ( Quercus robur L.) sacred plant associated with their highest god, Zeus or Jupiter the divinity of the sky and thunder. The olive tree ( Olea europaea L.) was a gift of Athena to the people of the city which is now named
Athens in her honor. Several plants, flowers and fruits are attributes of mythological Gods and Goddesses.
Demeter, Goddess of Agriculture is related with grains, especially wheat ( Triticum spp.) and the name 'cereals' is derived from Ceres, the Roman version of Demeter.
Vertumnus and Pomona (meaning fruit) are Roman protectors of fruit trees, gardens, and orchards, usually represented arraying a large platter of fruit or a cornucopia. Many botanists, such as Linné have named plants after mythological characters: Achillea L.
(Asteraceae), for Achilles Hero of Troy; Adonis L.
(Ranunculaceae) for Adonis, son of Mirra and lover of
Aphrodite; Artemisia L. (Asteraceae) for Artemisia =
Diana, Goddess of hunting; Asclepias Beos.
(Asclepiadaceae) for Asclepios = Aesculapius, famous healer; Centaurea L. (Asteraceae) and Centaurium L.
(Gentianaceae) for Centaur Chiron, the wiser Centaur;
Chloris L. (Poaceae) for Cloris or Cloe, nymph protectors of wild plants; Eragrostis (Poaceae), for Eros
= Cupid, God of love. The term hermaphrodite, used to describe bisexual flowers, derived from the tale of
Hermaphroditus, child of Aphrodite and Hermes, who was transformed into an androgynous being when united with the nymph Salmacis. Greek mythology and its
Roman version are rich in metamorphosis of characters into plants. Apollo myths are related with several transformations such as: Cyparissus, into cypress tree
( Cupressus ); the dryad Daphne into the laurel or bay tree
( Laurus nobilis L.), and Hyacinths' blood, into a flower whose petals have the inscription Ay Ay (Ai, Ai) as a symbol of his groan for his dead. The mythological
Hyacinth has been identified with a number of flowering plants other than the true hyacinth or iris. The flower mentioned by Ovid is equivalent to the flower that was transformed Ayax Telemonio and it is not Hyacinthus orientalis L. which was used in the pictorial representation of Hyacinth. The flower connecting both
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legends was identified as Consolida ajacis (L.) Schur.
(ex Delphinium ajacis L.).
Medicinal plants used in Thai traditional medicine: a case study in Kapchoeng Hospital, Surin Province,
Thailand
Chotchoungchatchai, S
1
Pornsiripongse, S 4
1, 2 , Saralamp, P
, Prathanturarug, S 2
2 , Jenjittikul, T 3 ,
MSc Program in Plant Science, Faculty of Science,
Mahidol University, Rajathevi, Thailand; Dept of
Pharmaceutical Botany, Faculty of Pharmacy, Mahidol
3
2
University, Thailand; Dept of Plant Science, Faculty of
Science, Mahidol University, Thailand; 4 Research
Institute for Languages and Cultures of Asia, Mahidol
University, Salaya, Thailand
Nowadays, Thai traditional medicine (TTM) is increasing in acceptance to be one of the alternative health care for Thai people, so TTM is available in many modern Thai hospitals. Although, many medicinal plants are promoted for increasing utilization in modern hospital, there have been still several problems such as lack of traditional use information, doubt in reliable sources of herbal material and limitation of integrating
TTM into modern hospital system. On the other hand, some modern hospitals are able to solve these problems and success in herbal used in modern hospital system.
One of these hospitals is Kapchoeng Hospital, a community hospital located in Surin Province, Thailand, where have routinely used herbal medicine since 1992.
Kapchoeng Hospital was selected to be a case study for documenting ethnobotanical uses of medicinal plants and to find out the factors affecting integration herbal used with TTM into modern hospital system. Fieldworks were carried out between August 2009 and October
2010. Data collection was done through semi-structure interview, participatory observation, plant specimen collection and botanical identification. The data has been analyzed and concluded by content analysis. The results show more than 116 medicinal plants were utilized through extemporaneous and combination preparations.
All herbal medicines were used for treatments of approximately 60 indications. Main source of herbal materials is from Cultivation and Collection Group.
Members of this group supply herbal material according the requirement of Kapchoeng Hospital in both quality and quantity. TTM practitioners are the specialists in herbal use at this hospital. They associate with modern medical staffs in health care team of Kapchoeng
Hospital. Not only TTM practitioners can use herbal medicine but also physician, pharmacist, nurse. There are
3 main factors for the success of herbal medicine utilization in Kapchoeng Hospital: knowledge and experience of herbal use with good results, respect of each profession and good relationship in health care team, and reliable source of herbal material from
Cultivation and Collection Group. From these results, the experience of herbal utilization in Kapchoeng Hospital could be useful not only for improve understanding of herbal used in modern hospital system, but also policy-
360 making in integrating the TTM into the health care system in Thailand.
Ethnobotanical investigations of Alamut region (Iran)
Ahvazi, M 1 , Khalighi-Sigaroodi, F
Charkhchian, M-M 4 , Mozaffarian, V 4
2 , Zakeri, H 3
, Mojab, F 5
,
2
1 Iranian Academic Center for Education, Dept of Culture and Research Herbarium (ACECR), Karaj, Iran;
Institute of Medicinal Plants, Iranian Academic Center for Education, Dept of Pharmacognosy and
3 Cell Line Pharmaceutics (ACECR), Karaj, Iran;
Engineering, Sigma Aldrich Biotechnology Division, St
4 Louis, USA; Research Institute of Forests and
Rangeland, Dept of Herbarium, Ghazvin, Iran; 5 School of Pharmacy and Pharmaceutical Sciences Research
Center, Shahid Beheshti University of Medical Sciences,
Tehran, Iran
We will present our results of an investigation on the ethnobotany of Alamut (Iran). Alamut is located northeast of the Ghazvin province in northwestern Iran.
Alamut is bordered to the north by the Mazandaran province and to the east by Tehran province. The climate is cold and humid with a 10 year average rainfall around
368.03 mm/year. In this region medicinal plants are often accessible health care alternative for most of the population in the rural areas and in fact folk herbal medicine is the most commonly used remedy to cure common diseases. Information on each of these plants was obtained by interviewing the native people, mainly the elderly and housewives. The collected plants, indicated by the locals, have been identified according to botanical references such as Flora Iranica . The exsiccate specimens of these plants are preserved at the Institute of
Medicinal Plants Herbarium (IMPH). We have identified
57 medicinal plants belong to 29 families used traditionally in Alamut’s rural areas. In this paper we describe these plants’ specifications and their application by native people.
Traditional medication in Tapkeshwari Hill Range of
1
Bhuj Taluka, Kachchh district, Gujarat (India)
Jain, B 1
MG Science Institute, India
Present study communicates the documentation of 38 medicinal plant species used for indigenous medication by the local villagers including Maldharies (pastoralists) and farmers of Tapkeshwari Hill Range of Kachchh district, Gujarat, India. Traditional knowledge on medicinally important plant species was collected from local villagers through a questionnaire survey using an open ended questionnaire data sheet. The response from the people interviewed clearly indicated that all the villagers were partially dependent on the forest for their daily health care requirement. Out of a total of 448 plant species recorded from the hill range, villagers of the selected villages used 209 species to cure various diseases. There were only 38 species which were commonly used by all villagers. Various plant parts like leaves, roots, seeds, bark, fruits, gum, flowers, latex and spadix paste of these species were used by local people to cure various diseases like dysentery, diabetes, skin

diseases, hypothermia, bronchitis, piles, snakebites, dropsy and fistula, cough and cold, ulcer, earache, gastric trouble, toothache, fever, indigestion, bile and diarrhoea.
Based on the field exposure, the most predominantly used ten plant species in order of preference (based on the preference of locals indicated by number of respondents) include Asparagus racemosus ,
Commiphora wightii , Capparis cartilaginea , Enicostema axillare , Fagonia schweienfurthii , Tinospora cordifolia ,
Balanites aegyptiaca , Cassia auriculata , Maytenus emerginata and Tribulus terrestris . The present paper describes the uses of 38 plant species in different ailments along with their local names, mode of preparation and dosage.
Database on anti-diabetic medicinal plants (DADMP) used by Valaiyans of Alagarkoil, Tamilnadu, India
Ramaraj, J 1 , Subramanian, R 1
Muthusamy, K 2
, Krishnasamy, G 2
1 RD Government Arts College, Tamilnadu, India; 2
,
Dept of Bioinformatics, Alagappa University, Tamilnadu,
India
WHO depicts that over 80% of world’s population depends on natural diversity and traditional system of medicine for healthcare needs (WHO, 2000). India, traditional medical practices are empirical in nature; however, an estimate suggests that over 200 million people with limited access to the primary healthcare services still depend on traditional medicine for medicine use plants to treat ailments. Surprisingly, many of the tribal and local communes in the rural area exploit the therapeutic potential of common plants that have not received much attention as their therapeutic potential is less known or have been scientifically unexplored. India possesses a total of 427 tribal communities with splendid diversity of indigenous tradition. The knowledge base and the practice have been marginalized due to political, social and economical reasons. Valaiyans of Alagarkoil are known to use more than 100 of plants as source to treat diabetics. Since in depth study on the usage of medicinal plants by Valaiyans of Alagarkoil Hills is far lacking, an attempt will be made to fill this gap. Anti-
Diabetic Medicinal Plants diversity informatics at present is not at instant access, data on anti-diabetic medicinal plants diversity either scanty in existing databases or other media not amenable to interactive search. Moreover majority of the literature do not contain photochemical informatics, ethnobotanical information, local names, in key words that make the computer search much more complicate. Literature survey on anti-diabetic medicinal plants diversity suggest that vast amount of information may be available in the scattered literature and has not been digitized. Therefore in the present study an attempt has been made to create a database for anti-diabetic medicinal plants diversity informatics. Database Anti-
Diabetic Medicinal Plants diversity (DADMP) has been systematically designed by cataloguing the information available on Anti-Diabetic Medicinal Plants and linked to various National and international databases (NCBI
Entrez, EMBL-EBI, Swiss-Prot, OpenIsis and Google) that provides added information on anti-diabetic healthcare needs. In recent times, mounting body of research is hoisting the credibility of underexplored traditional knowledge in meeting the challenges of primary healthcare services globally to treat diseases like medicinal plants. DADMP expected to serve the academicians, researchers and conservationists to exploit medicinal anti-diabetic medicinal plants in a sustainable manner for ethnopharmacological purposes. diabetics. AYUSH and other Indian folklore system of
Theme Poster numbers
01: Ecology, environmental change and conservation 0002 – 0236
02: Economic botany including biotechnology, agriculture and plant breeding 0237 – 0359
03: Genetics, genomics and bioinformatics
04: Physiology and biochemistry
0361 – 0457
0458 – 0607, 1098
05: Structure, development and cellular biology
06: Systematics, evolution, biogeography and biodiversity informatics
07: Plants in society
P0002 – ePoster
Response of peripheral population to extreme climatic events: results from a ten-year monitoring in the N-Apennines (Italy)
Abeli, T 1 , Gentili, R 2 , Rossi, G 1 , Cristofanelli, P 3
0609 – 0707
0708 – 1078, 1099, 1100
1079 – 1097
1 University of Pavia, Italy; 2 University of Milan-Bicocca,
Italy; 3 ISAC-CNR (Institute of Atmospheric Sciences and
Climate, National Research Council), Bologna, Italy
Peripheral plant populations tend to occur in less suitable environments and are often small, isolated and with low genetic variability with respect to central populations.
For these reasons they are expected to be more prone to extinction than central ones, due to stochastic events.
Climate change has been leading to a higher frequency of extreme climatic events that can represent a threat for peripheral populations of alpine/mountain species as a consequence of altered plant life attributes like phenology, reproduction, resource allocation, etc. We
361

studied the relationships between reproductive success of isolated populations of five arctic-alpine and orophitic species at the southern boundary of their range,
( Alopecurus alpinus , Carex foetida , Leucanthemopsis alpina , Silene suecica , Senecio incanus ) and climatic parameters over an 11 years period (1999–2009). We analysed the response of species reproductive performance during extreme climatic events. The study was performed in the Northern Apennines (N-Italy), an area predicted to be highly sensitive to climate change and have strong plant extinction events in the future, due to latitude and low altitude (<2200m). Species reproductive success was measured as number of flowering stems (NFS) and flowers/flowering stem (FFS) and % variation of NFS and FFS with respect to the previous year (FP and f/f, respectively). Mean temperature of May, June, July, August were obtained from the regional GAW station 'O. Vittori' at Mt. Cimone
(44°12'N, 10°42'E, 2165m a.s.l.) not far from the study site. The snow cover surface was recorded from 1999 to
2009, every 15 days starting from the 1st June until the
15th July. Redundancy Analysis of climate parameters and species fitness yielded significant relationships with
June temperature. C. foetida and S. incanus FP were distributed along an increasing snow cover gradient. S. incanus showed an increasing trend in NFS in 11 years.
We did not find any relationships between temperatures and NFS. Linear regressions showed negative and positive relationships between FP of A. alpinus and S. suecica , respectively and temperature variation of June.
FP of C.
foetida was positively related to temperature variation of May. L. alpina showed fluctuations in NFS in alternate years, probably due to resource limitation. In
2003, (extreme year due to the occurrence of summer heat-waves) NFS always decreased, reaching very low values, except for L. alpina and S.
suecica . Flowering decrease are followed by peak flowering the next year. f/f slightly varied from the mean value for S. incanus and S. suecica . FFS and f/f were not related to climatic data, for both S.
incanus and S. suecica . Extreme climatic events and resource limitation have detrimental effects on reproductive success of peripheral plant populations.
Despite the strong effect of climatic parameters, our peripheral populations seems to be stable or increasing in time, probably as a consequence of their ability to save resources during 'extreme' years to be relocate in more favourable years. Data showed that the effect of extreme years on reproductive success are visible one or two years after the event as an unusual increase in flower production for most of the species.
P0005 – ePoster
Eco-physiologic studies on some endemic plants living on serpentine at Sandras Mountain (Mula-Turkey)
Lu, Y 1
1 Ege Üniversity, Turkey
In Turkey, there has already been special interest in Ni accumulation by species of Aethionema R.Br., Alyssum
L., Bornmuellera Hausskn., Cochlearia L. and Thlaspi L.
(Brassicaceae). We now report instances of hyperaccumulation of Ni in some of the Turkish serpentine occurrences of Centaurea L. (Asteraceae).
There is worldwide interest in exploiting the property of
362 hyperaccumulation, both for remediation of metalcontaminated soils (phytoremediation) and for economic selective extraction of metal compounds by cropping hyperaccumulators (phytomining). The potential for these processes to be carried out in Turkey, the need for further exploration of the natural resource and the conservation issues involved are discussed. In this study, revealed in the relationship between Rosularia serpentinica (Werdermann) Muirhead, Teucrium sandrasicum O.Schwarz (which are serpentine endemic plants) and serpentine soils or serpentine rocks.
Observation area is Sandras Mountain of Mu ğ la (Turkey) from southwest Anatolia. This area has a large mass of serpentine, also serpentine has high amounts of heavy metal content. Contents of plant and soil samples were collected by using the Atomik Absorbsiyon spektrofotometre (AAS). In the end of the our analyses, soils the habitats of Rosularia serpentinica
(Werdermann) Muirhead and Teucrium sandrasicum O.
Schwarz are found to have high Ni, Cr, Mn, and Fe concentarations, However, very low rate of Mg/Ca are showed the negatives affected at plant growing.
P0007 – ePoster
Are subordinate ants the best seed dispersers?
Linking dominance hierarchy to seed dispersal ability in myrmecochory interactions
Aranda-Rickert, A 1
1 CRILAR-CONICET, Argentina
True myrmecochory involves the dispersal by ants of elaisome-bearing seeds. Between the guild of ants that are attracted to these seeds, only a few of them will act as effective dispersers, i.e., transporting the seeds to suitable sites (the nests) for germination and plant establishment.
Ant communities are known to be highly hierarchical, and competitively dominant ant species often occupy the most attractive food resources. In contrast, subordinate ants quickly deliver resources to their nests rather than consuming them on-site, thereby avoiding encounters with more dominant species. As a result, seed dispersal will be favoured when a subordinate ant first discovers a myrmecochorous seed. In this work, it is proposed that ant competitive hierarchies determine the ability of ant species to behave as effective dispersers of elaiosomebearing seeds, expecting that the best seed disperser ants will be subordinate species. Results from a series of studies carried out in semiarid northwestern Argentina found that Pogonomyrmex cunicularius Mayr. was the most subordinate species and also the main seed disperser of a myrmecochorous native shrub, whereas
Camponotus mus Roger was the most dominant species and the main elaiosome predator. Both the occurrence and the ability to first discover the baits were significantly influenced by the time period of the day. At midday the behaviourally dominant C. mus ants were virtually absent from baits, but instead monopolized them at more moderate temperatures. On the other hand,
P. cunicularius showed a higher occurrence and discovering ability at the highest temperatures of the day.
This strategy allowed P. cunicularius ants to avoid the risk of interference competition with dominant species.
In summary, the behaviour of subordinate species appears to be integral to successful myrmecochory, but

also the temperature at which the dispersal takes place plays a significant role by shaping the ant community interactions. Therefore, myrmecochorous plants should not only favour their discovery by subordinate ants, but also should present their seeds at those times of the day when behaviourally dominant ants are less active.
P0008 – ePoster
Woody community dynamics in a Brazilian savanna on rocky soils located at the Pireneus State Park,
Brazilian State of Goiás
Arcela, V 1 , Pinto, JRR 1
1 Brasília University (UnB), Brazil
The biome known as Cerrado, which contains the richest savanna in the world, is located in central Brazil. Cerrado rupestre (Brazilian savanna on rocky soils) is a sub-type of cerrado stricto sensu characterized by shallow soils and rock outcrops. Floristic studies in these environments are scarce and there is no information about the dynamics of these communities. The aim of this study was to evaluate changes in floristic and structural composition of the woody plant community of a cerrado rupestre located in Pireneus State Park (Goiás, Brazil), during a five-year period. Consecutive vegetation surveys (2004 and 2009) of the woody species (at least 5 cm of trunk diameter at 30 cm above the ground) were conducted in
1.0 hectare (ten plots of 20 × 50 m) of cerrado rupestre.
We sampled 70 species belonging to 55 genera and 31 families over the monitored time. Even under fire conditions inside the Park, there was an entrance of nine species, while three others disappeared in the same area.
The Shannon Wiener´s diversity and Pielou´s equability indices changed from 3.37 and 0.82 in 2004 to 3.39 and
0.81 in 2009, respectively. In both surveys, the distribution of individuals in diameter classes showed a reversed J-shape, without a meaningful statistical deviation. Changes in the ranking of the most representative species in the community were observed, although Schwartzia adamantium obtained the highest IV in both years. The community presented a 0.2% increase in basal area and a 3.4% decrease in density of individuals during the monitored period. The community´s mortality rate (8.4% yr the recruitment rate (8.2% yr -1
-1 ) was higher than
). The periodic annual increment (PAI) observed in the community was 0.23 cm yr -1 . The species with the highest PAI was Sclerolobium paniculatum (0.74). This community presented higher mortality and recruitment rates than those found in other
Brazilian vegetation types, showing how high the turnover of individuals in cerrado rupestre could be.
Nevertheless, this community showed higher PAI than those obtained by others studies. Periodic monitoring of cerrado rupestre should be continued to elucidate consistent information about possible floristic and structural changes that may occur in rocky soil environments. Keywords: floristic change, structural change, mortality rate, recruitment rate, cerrado rupestre
P0009 – ePoster
Carbohydrate metabolism of the Amazonian tree
Senna reticulata
under elevated CO
2
Arenque, B 1
Buckeridge, M 1
, Grandis, A 1 , Souza, A 1 , Pocius, O 1 ,
1 Universidade de São Paulo, Brazil
Plants of floodplains tend to show many morphological and physiological strategies to get higher survival rates in this places. One of them but with received less attention is the plasticity showed by carbohydrate metabolism that is directly linked to high energy demand at this sites.
Senna reticulata (Leguminosae) is a pioneer tree which is common in floodplains sites submitted to an annual pulse of flooding at Central Amazonian and one of the most efficient colonizers at this sites besides extremely flooding tolerant. In this work by the first time was made the characterization about reserves and mobilizations patterns of sugars on the time range that corresponding to the establishment at the initial first terrestrial phase.
Additionally, the effect of high CO
2
was checked by growing plants in Open-Top-Chambers (OTCs) with ambient (~380
µ mol) and elevated (~760
µ mol) CO
2 concentrations. It was made systematic harvests along periods of 24 hours and 90 days to determinate the carbohydrate non-structural content. It was founded that
S. reticulata had higher levels of starch in leaves showing evidences that this reserve is directly linked with constantly production of new leaves strategy. The ratio sucrose:monosaccharides was high in all organs but specially in roots showing evidences that this specie has metabolic strategies also related with drought periods because besides sucrose could be used as the aim substrate to adventitious root formation at flooding conditions and can be useful too on the osmotic regulation of roots in sites with low moisture content soil. The high levels of carbon dioxide promote significative increase of starch in leaves and stem besides decrease the degradation rate of transitory starch in leaves in the dark period. The levels of soluble sugars
(glucose, frutose and sucrose) showed tendency of decrease under high CO
2
conditions probably due to higher use in root growth. This set of answers evidence that this tree has positively response to high CO
2
useful by possible mechanisms that decrease the signaling through sugars like for example the big abiliity of increase capacity of preexistents sinks to promoting the surplus carbohydrates mobilization to reserves and growth.
P0010 – ePoster
Role of seedborne fungi on percentage germination of three varieties of castor (
Ricinus communis
L.) and their growth performance by incorporation of AM
1 fungi
Arya, A 1 , Misra, R 1
Maharaja Sayajirao University of Baroda, Vadodara,
India
India is one of the leading producers of castor. The oil of castor ( Ricinus communis L.) is used in pharmaceuticals, medicine and industries related to soap, paint, and lubricants etc. Agricultural scientists in the country have developed a large number of hybrid varieties exploiting hybrid vigor. The studies were undertaken to assess the occurrence of seed borne mycoflora at different
363

temperatures, when the seeds were stored for one year in different types of containers in Baroda, India. Seed health ensures a better crop in the field. Seed storage is a serious problem in tropical countries. The experimental results showed association of 19 different fungi with three different varieties of castor. Presence of seed mycoflora reduced percentage seed germination to 10 in GCH4, 20 in Avani 41, and 38 in local var. after storage at 250˚C. It was 13% in GCH4, while germination reduced to 7 and
12 % in Avani41 and local var. when stored to one year at 100˚C. The growth of plants is influenced by
Mycorrhizal Symbiosis. AM fungi not only provide nutrients and vital minerals necessary for growth but also provide water balance and protection from phytopathogens. To assess the role of AM fungi on growth enhancement a pot experiment was performed. In which AM consortium was added and increase in plant biomass and disease appearance was recorded. An increase in plant biomass was recorded in all the treatments. After 90 d of growth the number of leaves were 9 as compared to 8 in control plants of local var.
The leaves were 14 in other two var. after 90 d. Five percent wilting was recorded in control plants of local var., in other plants no such symptoms were recorded.
P0012 – ePoster
Use of remote sensing in the analysis of fructification phenology in a Pantanal mesoregion of Brazil
Atique, G 1 , Costa Penatti, N
Francis Roche, K 3
2 , de Melo Bandeira, H
, Paranhos Filho, A 3
2 ,
1 Post-Graduate Program in Vegetal Biology,
Universidade Estadual de Campinas, São Paulo, Brazil;
2 Applied Geotechnologies Laboratory, Dept of
Hydraulics and Transport, CCET, Universidade Federal
3 de Mato Grosso do Sul, Campo Grande, Brazil; Postgraduate Program in Ecology and Conservation,
Universidade Federal de Mato Grosso do Sul, Campo
Grande, Brazil
Phenological changes in plant populations are strongly related to environmental variation in such characteristics as water availability, temperature and solar radiation.
Phenological studies have traditionally been carried out through ground observations. However, in recent years, remote sensing (RS) using satellite-borne imaging systems has been widely adopted to monitor vegetation phenology. This paper aimed to compare the above indices, using Landsat TM images, procuring relationships with ground gathered data. The fruiting phenology of gallery forest plant communities of an area next to Brazilian Pantanal were studied. Specifically we hypothesized that the potential of the indices to reflect the changes in fructification phenology of the plant community studied. Ground data on the fruiting phenology were gathered, during the course of one year, and compared with Landsat TM images, with application of the following indices: NDVI (Normalized Difference
Vegetation Index), SAVI (Soil-adjusted Vegetation
Index), LAI (Leaf Area Index) and MVI (Moisture
Vegetation Index). Comparisons were made of the efficiencies of the different indices in detecting changes in fructification phenology. Values of all four indices decreased during the dry season. This could have been a consequence of the decreasing quantity of chlorophyll in
364 the plants during the height of the dry season, involving low water availability and decreasing photosynthetic rates. The fact that seasonal changes in air temperature were relatively small suggests that it was principally the water balance that limited the vegetation growth.
Because the numbers of species and individual plants bearing fruit in the plant community increased during this period, the VI’s were inversely proportional to these abundances. The lowest values of the VI’s, coincident with high production of anemochoric fruits, could also have been related to the loss of chlorophyll due to fruit maturation. The peak value for zoochoric fruits occurred in December, when increases in the four indices, manuscript especially the LAI, were recorded. The NDVI was the least accurate in reflecting the changes in vegetation phenology. The present study has shown that fructification phenology can be reflected by various vegetation indices. Landsat images were successfully used, and the relative effectiveness of different indices identified. These findings could have important implications for research in such areas as environmental conservation, the sustainable exploitation of natural plant communities, and climate change.
P0014 – ePoster
Army and DoD approaches to managing spread of invasive species
Balbach, H 1 , Rew, L 2 , Pollnac, F 2 , Brummer, T 2
1 US Army ERDC, Champaign, Illinois, USA;
State University, Bozeman, Montana, USA
2 Montana
Control of the movement of invasive plant species has long been a matter of grave concern to government agriculture agencies. Australia is one such concerned country, as are New Zealand, the United States, and many others. The level of concern and responses to the problem are widely varied, however. With respect to military equipment especially, the retrograde shipment of vehicles to the USA following the Gulf War in the 1990s heightened these concerns. This led to the creation of a
Presidential Special Oversight Board in 1999 to investigate, among other things, the capability to adequately clean returning military vehicles and equipment. Presidential Executive Order 13112, also from 1999, directed that all agencies monitor and minimize movement of non-indigenous invasive species.
The order has been interpreted in different ways by different agencies. Those with activities largely taking place within the U.S. assumed that it applied to those activities. The U.S. Forest Service, for example, some time ago implemented the requirement to wash vehicles entering and leaving forest fire management areas. They also developed a prototype portable vehicle washing system to perform these tasks. The Department of
Defense, however, sensitized by the issues relating to the return of equipment used in Operation Desert Storm in
1991, focused on the need to clean vehicles being retrograded from overseas. While these concerns were clearly the more critical ones, this has left the Army and
DoD without clear policy or guidance as it relates to the risks and preventive measures appropriate for preventing the spread of invasive species from place to place within the continental United States. It is clear that many elements of risk are present when vehicles, containers,

and especially construction equipment are transported from place to place within the country, as well as internationally. National Guard and Army Reserve components may often train at several locations in a year, and regular Army units may conduct joint exercises at other installations or on National Forest or Bureau of
Land Management public lands as well. Only a few locations require vehicles to be cleaned before entering.
A series of research efforts over several years has allowed the risks of different activities to be quantified.
One important aspect has been to determine how effective existing cleaning methodologies are in removing seeds and other propagules from the vehicles.
Another focus has been to quantify the potential for different vehicles to pick up weed seeds when they are operated either on or off road. In one study, it was found that existing vehicle cleaning using field-portable equipment typically removes about 80% of the soil on a vehicle. This clearly reduces risk of transport from place to place, but does not eliminate it. The data and risk assessments are applicable to all types of vehicles and all public and private operators of the vehicles, including the
Forest Service, Army, private ranchers, and the recreational public.
P0015 – ePoster
Self-incompatibility, pollen limitation, and endangerment in the Hawaiian silversword alliance
(Compositae) on Kaua`i
Bainbridge, SJ 1 , Baldwin, BG 2
1 Jepson Herbarium, Berkeley, USA;
Biology, Berkeley, USA
2 Dept of Integrative
Strong self-incompatibility in the Hawaiian silversword alliance is highly unusual for angiosperms of remote oceanic islands and makes these plants especially vulnerable to severe pollen limitation from pollinator loss, loss of S-allele diversity, or both. In wet forests of
Kaua`i, lack of recruitment of the locally common tree
Dubautia knudsenii subsp. knudsenii and extreme rarity of the shrub or small tree D. kalalauensis and the liana D. latifolia led us to study whether pollen limitation impacts these taxa and, if so, why. Controlled crosses in each taxon between plants of the same or different stands and controlled self-pollinations were conducted over consecutive years (2007 and 2008) and stands were surveyed for any naturally recruited seedlings or juveniles. Results of these hand pollinations in comparison with levels of natural seed set of the same plants indicated that D. knudsenii subsp. knudsenii was completely pollen limited, with no natural seed set detected. Extensive seed set and germination of vigorous seedlings from within- and between-stand hand-crosses of D. knudsenii subsp. knudsenii indicated that lack of pollinator service and not loss of S-allele diversity (or other genetic factors) was responsible for lack of natural seed set. Absence of any observed visitation by potential pollinators, discovery of only one juvenile plant across both studied stands, and unusual floral characteristics
(including minimal pollen production) of D. knudsenii subsp. knudsenii raise concern that the taxon may have lost its pollinator and be biologically extinct despite still being locally common. In contrast, results from the sympatric D. kalalauensis indicate minimal pollen limitation, substantial recruitment, and extensive visitation by potential pollinators, with active destruction of entire plants by feral goats and, apparently, by invasive Passiflora mollissima . Reproductive failure in natural and within-stand crosses of the rare D. latifolia appear to implicate small population sizes and lack of within-stand cross-compatibility as major contributors to endangerment. Preliminary results from other taxa of
Dubautia from Kaua`i reinforce the above finding that different factors can be most important as causes of rarity or endangerment of closely related taxa sharing the same environment.
P0018 – ePoster
Importance of conservation units in the Atlantic
Forest, southeastern Brazil, to the Orchidaceae family
Baumgratz, JFA 1 ,Barberena, FFVA 2 , Barros, F 3
1 Museu Nacional/Pós-graduação, Universidade Federal do Rio de Janeiro, Brazil; 2 Instituto de Pesquisas Jardim
Botânico do Rio de Janeiro, Brazil; 3 Instituto de
Botânica, Estado de São Paulo, Brazil
The Atlantic Forest biome is the second largest forest formation in the Neotropics, and one of the most threatened biomes on the planet. This biome has high degree of endemism and richness species, and 92% of this biome is located in Brazil along the northeastern and southeastern coast. However, the Conservation Units protect only 9% of Brazilian forest remnants and only
0.03% of the forest fragments are larger than 10,000 ha.
The conservation of the larger forest fragments and in advanced stage of succession is a strategic priority. In this context, it can be mentioned the National Park of
Itatiaia, in the states of Rio de Janeiro and Minas Gerais, southeastern Brazil. This Conservation Unit has ca.
30,000 ha, is located at altitudes between 500 and 2,787 m, and is covered by montane forests and high altitude grasslands. Brazil is known as a megadiverse country in
Orchidaceae, with more than 2,650 species, of which more than 50% occur in the Atlantic Forest. This study shows the importance of National Park of Itatiaia in the preservation of Orchidaceae in the Atlantic Forest, describing the diversity, endemism, and conservation data. This study was based on literature data, analysis of herbaria collections and expeditions to the area during two years for collecting botanical material. In the area occur 85 genera and 226 taxa, which represent ca. 9% of
Orchidaceae in Brazil. 108 taxa are endemic of Atlantic
Forest, of which 98 are also endemic to the country, and two species ( Acianthera capanemae (Barb.Rodr.)
Pridgeon & M.W.Chase, Octomeria itatiaiae Brade &
Pabst) are endemic to the state of Rio de Janeiro.
Octomeria itatiaiae Brade & Pabst is the only species mentioned in the list of rare species from Brazil, and known only by the type. Five species are exclusive of the high altitude grasslands: Coppensia caldensis (Rchb.f.)
Docha Neto, Habenaria itatiayae Schltr., Habenaria rolfeana Schltr., Pelexia itatiaiae Schltr., and Prescottia phleoides Lindl. In addition, 46 species appear in the list of threatened species of southeastern Brazil with different degrees of threat, with 50% classified as
Vulnerable. Thus, the maintenance of Conservation Units in the Atlantic Forest is an important conservation
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practice and relevant to the preservation of the
Orchidaceae species and the environment.
P0020 – ePoster
Mycorrhizal fungi in
Cattleya
(Orchidaceae) species occuring in Atlantic Forest and Cerrado, Brazil
Bocayuva, M 1 , Oliveira, S
, Otoni, W 3
1 , Veloso, T 1
, Kasuya, M 1
, Ribeiro, L 1 ,
1
Liparini, O 2
Laboratory of Mycorrhizal Association, Dept of
Microbiology, BIOAGRO, Federal University of Viçosa
(UFV), Viçosa, Brazil; 2 Dept Plant Pathology, UFV,
Viçosa, Brazil; 3 Dept Plant Biology, UFV, Viçosa, Brazil
The current knowledge of threatened orchid species in
Brazil is still very incipient not only due to the lack of specific studies, but also to the megabiodiversity of orchid species. The official Redlist of Brazilian flora considers around 356 vascular plant species, which includes 34 representatives of Orchidaceae. Among
25,000 natural species distributed throughout the world,
1779 are included in any conservation status, which represents 14 % of them are endangered species. Brazil presents 2,400 species of orchids, with few data related to the conservation status of natural populations. Due to the fact that the distribution of these species concentrates on the Atlantic Forest, specially on the southeast region, a hotspot with less than 10 % of its original ranging, it becomes essential to study the natural populations as well as ex situ conservation techniques. Some of the orchid species selected for this study are mentioned on the Redlist of Brazilian flora and occur mainly in
Atlantic Forest and Cerrado areas of the Minas Gerais and Rio de Janeiro States. Cattleya jongheana (Rchb.f.)
Van den Berg and C. perrinii Lindl., are epiphytic species with high ornamental value and with a restricted geographic range. As well, Cattleya caulescens (Lindl.)
Van den Berg, a Campo Rupestre terrestrial orchid is found in large populations in private areas of iron mine at
Minas Gerais. Cattleya cinnabarina (Bateman ex Lindl.)
Van den Berg also occurs in Campo Rupestre area and, at the moment, it is the only species studied not cited on the
Redlist. The major objective is to establish a method of in vitro propagation of these four orchid species and to evaluate the biodiversity of mycorrhizal fungi associated with these orchids. Thus, the strategies to be used to achieve this goal are: (a) to identify morphologically and molecularly mycorrhizal fungi associated with orchids; and (b) to evaluate methods of in vitro propagation and acclimatization of orchids grown in laboratory. To date, a variety of binucleate isolates, probably of the genus
Epulorhiza were obtained with different colony diameter, colour, border and aerial mycelia aspect. DGGE analyzes have been conducted to confirm this fungi biodiversity.
Seeds of orchid species were inoculated, individually, using isolates of Epulorhiza and incubated in light.
Stages of development were performed after 3 and 10 weeks of inoculation and also for control treatment
(OMA and Knudson). After 3 weeks, in the symbiotic culture, the development stage of elongation of first leaf was observed, against the major stage in the uninoculated control that was the appearance of the protomeristem.
Our results will contribute to the knowledge of the biodiversity and ecology of orchids, mycorrhizal fungi and therefore the habitat. Finally, studies with emphasis
366 on conservation can also assist in the management of impacted areas and encourage the creation of groups of environmental education to work with the local population. (Support: AOS,CNPq,FAPEMIG)
P0021 – ePoster
Effect of mother plant age on germination and size of seeds and seedlings in the perennial plant
Bogdanowicz, AM 1 , Lembicz, M 1 , Zukowski, W 1
1 Adam Mickiewicz University, Dept of Plant Taxonomy,
Poland
The performance of seeds and seedlings in relation to the age of the mother plant was studied in Carex secalina .
Seeds of this sedge can differ substantially in size. We planted 100 C. secalina individuals from three populations in a common garden and followed them for four years. We found that mean seed mass varied with plant age, but the pattern of variation was populationspecific, with only one population showing significant reduction in seed mass with age. Similarly, germination rate changed with age differently in different populations.
The relationship between the age of the mother plant and the length of emerged seedlings did not differ between populations. In spite of the fact that plant size and mean seed mass exhibited similar patterns of variation within populations, there was no correlation between these two variables at the level of individuals. This means that the size of C. secalina tufts does not determine how big the seeds will be. Moreover, there was no relationship between mean seed mass and the length of seedlings.
Presumably, factors intrinsic to each plant determine the production of either small or large seeds in a populationspecific way.
P0022 – ePoster
Assessing air quality of public parks in Bangkok,
Thailand from photosynthesis and chlorophyll fluorescence of the transplanted lichen
Parmotrema
1 tinctorum
(Nyl.) Hale
Boonpeng, C 1 , Boonpragob, K 1
Ramkhamhaeng University, Thailand
Lichens are recognized as effective biomonitor of air pollution. The objective of this study was to use lichen to assess air quality of public parks in Bangkok by observing photosynthesis and chlorophyll fluorescence of transplanted lichen. Parmotrema tinctorum (Nyl.) Hale was transplanted from Khao Yai National Park, the unpolluted area, to ten public parks in Bangkok.
Photosynthesis and chlorophyll fluorescence of the lichens were measured before and at different time intervals after transplantation. The results showed that photosynthesis of the transplanted lichens declined to
15–60% of the pre-transplanted value after 45 days of exposing to urban air, whereas Fv/Fm varied to 10–33% of the original condition. Transplanted lichens in the city center were seriously affected than those in the sub urban areas. The study demonstrated that lichens can be used to assess air quality of public parks in Bangkok.

P0023 – ePoster
Hydraulic redistribution of fog water from shoots to the rhizosphere in the cloud forest species,
Drimys brasiliensis
Miers.
Breder Eller, C 1 , Silva Oliveira, R 1
1 UNICAMP, Brazil
Fog is an important source of water for plants in tropical cloud forests but the mechanisms by which plants use fog water are still unclear. Here we investigated the occurrence of hydraulic redistribution (HR) of fog water from shoots to the rhizosphere during nocturnal fog events propelled by water potential gradients between a saturated atmosphere and dry soil. We used the heat ratio method to monitor bi-directional sap flow in the stems of a common Neotropical cloud forest species, Drimys brasiliensis Miers (Winteraceae), in a greenhouse experiment where saplings were subjected to nocturnal nebulizations and soil drought. Additionally, we monitored the soil water content of the potted plants to determine if fog water was actually released into the rhizosphere. We also measured maximum photosynthesis
(A) and daily stomatal conductance (gs) to evaluate the influence of fog in the ecophysiological performance of the saplings. We prevented fog drip to reach the soil by placing plastic bags on each pot. Nocturnal nebulizations led to sap flow reversals and increased the soil water content above control plants. Sap flow reversals represented up to 12% of the maximum diurnal sap flow velocities. Plants subjected to crown nebulizations also had higher A and gs than plants subjected to drought
(p<0.05). The redistribution of fog water mediated by the crowns of D. brasiliensis to their rhizosphere may allow these plants to photosynthesize and transpire during periods of seasonal droughts. Therefore the HR of fog water may play an important role in the ecohydrology of cloud forests subjected to soil drought.
P0024 – ePoster
Does pollen production translate into photosynthetic differences between female and male
Gaultheria poeppigii
plants?
Briceno, V 1 , Cruz, M 2
A 1
, Ostria, E 3 , Corcuera, L 3 , Nicotra,
1
2
Australian National University, Canberra, Australia;
3
Universidad Nacional de Colombia, Colombia;
Universidad de Concepcion, Chile
Gaultheria poeppigii is an evergreen dioecious shrub endemic to Chile and Argentina. The sexes are easily identified when flowering, as males produce large amount of pollen whereas females produce fruits and seeds. Hence, the cost of reproduction for female and male plants is different. Male plants are predicted to have larger energy expenses during flowering, and females are expected to have overll greater costs when fruit production is considered. Here, we focused on the flowering period and we ask whether the cost of pollen production is translated into photosynthetic differences between male and female individuals. We hypothesized that pollen production is expensive in terms of N, which might be translocated from leaves to pollen causing a decrease of photosynthetic capacity in the leaves nearest to the flowers. To test that, we selected fifteen females and fifteen males in Katalapi Park (X Region de Los
Lagos, Puerto Montt, South of Chile). In these plants we chose flowering branches which had two leaf cohorts, one year old (new leaves) and two years old (old leaves) and flowers located in between. We measured chlorophyll fluorescence and net photosynthesis (Pn) at the same time. In addition, light curves of fluorescence were measured in a Dual-Pam Fluorometer, so that we could study the PSI and PSII performance simultaneously. Furthermore, old leaves of both sexes were exposed to high light intensity for 3 hours to see if photoinhibition occurred to the same extent in both sexes. Chlorophyll content was also measured. We found that new female leaves had the lowest net photosynthesis compared with all other leaf types and also they have lower electron transport rate of PSI (ETR I) and yield of photosystem I (YI) compared with older females leaves.
There were no differences between the sexes in non photochemical quenching (NPQ) or electron transport rate of photosysthem II (ETRII). In both sexes new leaves had higher NPQ and lower ETRII compared with older leaves. We did not find evidence of photoinhibition in any treatment and no sexual differences in chlorophyll were found between sexes, nevertheless new leaves have lower chlorophyll values. These data do not show a decrease in photosynthesis in male plants associated with pollen production in either new or old leaves, in fact male new leaves have higher net photosynthesis than the new female leaves. It is likely therefore that leaf N is not being reallocated from leaves for pollen production; instead male new leaves up-regulate photosynthesis to pay for the cost of reproduction.
P0025 – ePoster
Global biogeographic pattern of floral ovule number: is there an Ashmole's Rule for angiosperms?
1
Burd, M 1
Monash University, Victoria, Australia
Ovule number per flower bears some functional similarities to clutch size in birds. Clutch sizes show strong biogeographic structure (increasing egg production with increasing lattitude) both within and among bird species. A leading explanation for this pattern is known as Ashmole's Hypothesis, and concerns the effect of seasonality at high latitudes on mortality rates and per capita food availability. What about flowering plants? Data from 187 species suggests that any pattern in floral ovule number runs opposite to that for bird clutch size: ovule number tends to be lower at higher latitudes. I propose an explanation for this based on previous work relating ovule number to stochastic variation in pollination success. If correct, this explanation would also suggest an alternative to
Ashmole's hypothesis for the biogeography of clutch size in birds. Australia may provide a special opportunity to test ecological explanations for the evolution of floral ovule number, in that ecological variability in Australia tends to be more independent of latitude than in other continents.
367

P0026 – ePoster
So near, yet so far: evolutionary relationships among rare
Tetratheca
species endemic to banded ironstone ranges in Western Australia.
Butcher, R 1 , Byrne, M 1 , Crayn, D 2
1 Science Division, Dept of Environment and
Conservation, Perth, Australia; 2 Australian Tropical
Herbarium, James Cook University Cairns Campus,
Australia
The banded iron formation (BIF) ranges scattered across
Western Australia's Yilgarn Craton are small, ancient, biodiverse and highly prospective for iron-ore mining.
Driven by development threats, morphological and molecular studies were undertaken to assess the taxonomic status and evolutionary relationships of three
Declared Rare species of 'leafless' Tetratheca endemic to three adjacent BIF ranges in the Goldfields, as well as the positions of three, putatively new, morphologically similar taxa. These investigations confirmed the distinctness of the rare species T. aphylla F.Muell., T. harperi F.Muell. and T. paynterae Alford and identified one new rare species and two new rare subspecies from collections affiliated with T. aphylla and T. paynterae .
The recognition of these taxa at specific and subspecific ranks was based on their different degrees of morphological and molecular divergence, combined with geographic disjunction. Cladistic analysis of nrDNA ITS and cpDNA trnL-trnF sequences from a range of
Tetratheca species from Western Australia and the eastern States indicates that T. aphylla , T. harperi and T. paynterae belong to three separate evolutionary lineages and that the endemism displayed among these taxa to small, disjunct ranges within the same geographic area, is a result of in situ speciation due to historical fragmentation. These results exemplify the extremely high conservation value of the Goldfields BIF ranges, where another new, rare species of Tetratheca allied to T. harperi has since been located and described. The superficial similarity among the study taxa in having a
'leafless' habit can be seen to be adaptive convergence in response to the marginal and semi-arid environments in which they occur, and this character is highly homoplastic within the genus.
P0027 – ePoster
Guyana's avoided deforestation climate policy opportunities for post-2010 biodiversity targets: the
Botanical Dimension
Caesar, JC 1
1 University of Guyana, Guyana
In the Anthropocene Age, accelerating biodiversity loss is intertwined with the overall threats to human security, human well-being and earth system ecological sustainability from global climate change. This potential reality has engendered serious evidence-based policymaking globally. Guyana has taken bold steps in articulating a national climate adaptation and mitigation policy within the context of a Low Carbon Development
Strategy (LCDS), largely as a consequence of Guyana's
368 firsthand experience with increased frequency and intensity of climate change-induced flash floods disasters and notable vulnerability to sea-level rise. The nation advocates avoided deforestation (REDD+) as a prudent and practical mitigation and adaptation strategy. The predominantly coastal population has left vast expanses of forest reasonably free from serious threats. Despite the paucity of local taxonomic capacity generally, significant work has been achieved on higher botanical biodiversity taxa across selected areas of the country's geopolitical boundaries. We believe avoided deforestation provides an excellent opportunity for marrying Guyana's climate change mitigation policy with collateral benefits of working towards post-2010 biodiversity targets in general, and especially, the botanical dimension for a number of pristine areas not yet significantly inventoried in tandem with biodiversity loss mitigation. The pressure on natural resource use and land use management issues from gold mining and other alternative interests require an aggressive national taxonomy strategy to fully assess the nation's botanical wealth within the broad framework of the Low Carbon REDD+ strategy. We discuss a framework strategy emerging from these REDD+ opportunities for promoting biodiversity inventories generally and botanical diversity inventories in particular, across all national terrestrial, aquatic and marine ecosystems, with particular emphasis on endemic species and less studied plant families. Ecological studies, germplasm collections, gene banking, conservation genetics of important forest species, and climate impact ecophysiological studies may need equal prominence in the overall strategic framework within resource allocation priorities of the LCDS. Guyana's largely untapped botanical diversity can provide the most useful and sustainable natural capital for expanding value added botanical products and related R&D innovations while mitigating climate change, creating jobs, reducing poverty, meeting the Millennium Development Goals, alleviating the environmental drivers of biodiversity loss and ecosystem degradation, and enhancing human wellbeing especially of indigenous communities under the
REDD+ framework. This provides a perfect opportunity for marrying an enhanced and more effective participation in the Global Taxonomy Initiative with national biodiversity conservation priorities and Guyana's contribution and obligations to the Global Plant
Conservation Strategy and the Convention on Biological
Diversity.
P0028 – ePoster
Indigenous people’s cultural beliefs and plant biodiversity conservation in the climate change era
1 ecology policy framework
Caesar, JC 1
University of Guyana, Guyana
Cultural beliefs among our indigenous relatives may represent opportunities for unique systems of ethnoconservation strategies to safeguard forest ecosystems. The roles of cosmology, taboos, folklore, spiritualism, religion, ethnobiology, among others, together provide a holistic view of the role of indigenous knowledge in ecologically sustainable development and the maintenance of the ecological integrity of the

biosphere especially in the climate change era. Cultural diversity is an important socio-ecological construct. On face-value, indigenous knowledge may be erroneously perceived as primitive attitudes, lifestyles, thinking, and beliefs. This poster briefly reviews corollaries of forest dwellers’ beliefs in the mystique of the forest and its biodiversity and how best we can adapt and formulate some of these cultural diversity socio-ecological constructs into adaptive management paradigms for meeting the needs of climate and environmental justice for forest governance under the REDD+ mechanism for climate change mitigation.
P0029 – ePoster
Effects of UV and temperature on
Sphagnum
biology
Cardona-Correa, C 1 , Graham, L 1 , Graham, J 1
1 University of Wisconsin, Madison, USA
During the past decade or so ozone (O
3
) layer depletion, particularly at high latitudes, has become an ecological concern. Studies conducted by The World Health
Organization have indicated that the rate of O
3
depletion has leveled off. Even so, experts point out that the damage caused by chlorofluorocarbons (CFCs) has been so great that a time period of ~50–60 years (2050–2060) will be needed for ozone to recover enough for surface
UV to return to 1980 levels. For this reason it is important to evaluate the impact of UV radiation on high latitude organisms that could be affected during the recovery period. The peat moss Sphagnum plays an important role in modern biogeochemical systems, especially in north temperate regions, where it dominates an estimated 3% of global landmass and stores approximately 400 gigatons of C peatmoss abundance, photosynthetic activities, and production of cell wallbound hydrolysis-resistant polymers that shield UV radiation and help to sequester organic C are all important factors in C-cycling on a global level. Previous research indicates that UV radiation impacts peat moss photosynthesis and pigment pool size, but not total biomass. To aid the prediction of future UV and global warming impacts on peat mosses and their carbon storage capacity, we evaluated the combined effects of temperature and UV exposure on the production of resistant C by Sphagnum compactum , We selected this species to represent Northern Hemisphere peat mosses because its relatively small size facilitates laboratory experimentation, as in our previous demonstration of mixotrophy (Graham et al. 2010). Here, we report the results of studies of laboratory cultures of S. compactum concurrently exposed to three temperature regimes (8–
10°C, 20°C, 28–30°C) and two UV radiation levels in a full factorial experimental array, while maintaining constant PAR. Acetolysis was performed to provide a conservative estimate of percent resistant C produced in each experimental condition. Preliminary data demonstrate that at 20°C, Sphagnum sequesters less C than when grown at 10°C, and that plants exposed to enhanced UV at the higher T exhibited rapid death, bleached biomass indicating severe impact on the photosynthetic apparatus. These results suggest that global warming may exacerbate UV impacts on high latitude peatmosses, thereby reducing their ability to sequester C.
P0030 – ePoster
Distribution, ecology, demography and conservation of rare and endangered
Limonium
Mill. species
(Plumbaginaceae) in the Mediterranean
Caruso, G 1 , Uzunov, D 1 , Bogdanoviæ, S 2
1 Dept of Environmental and Crop Science, Marche
Polytechnic University, Italy; 2 Dept of Botany, Faculty of
Science, University of Zagreb, Croatia
The genus Limonium Mill. (Plumbaginaceae) in the
Mediterranean area is a taxon with active speciation processes, including many microspecies with high conservation value. The present study deals with halophyte, chasmophyte, narrow endemics occurring on the rocky coast of Calabria (S Italy): Limonium lacinium
Arrigoni, L. calabrum Brullo and L. brutium Brullo. In order to evaluate the general trend of the distribution, as part of historic biogeographical relations and future evolution, the approach of the asymmetric areale analysis is applied. It implies that the coast belt where the plant lives could be represented as a line running parallel to coast profile. Along this line the occurrence of the studied species is checked and censused. Starting from presence/absence analysis the population density and anisotropy (of natural or anthropic origin) are calculated, and the most relevant stands, in a conservation perspective, are identified, as well as the main threat factors. Besides applying GIS techniques the correlation between local population structure and different biotic and abiotic factors, micro-chorology, cliff geomorphology, etc. are evaluated. Different models for the ecological features of the three species, related to the main environmental typologies, are critically evaluated and a programme for long term monitoring is proposed introducing further ecological investigation (e.g. reproductive biology, colonization strategies, etc.). Some of these species are included in the regional Red List of rare and threatened plants. For the assessment of the global conservation status the IUCN methodology is applied. On the base of the present study a more efficient strategy for in situ and ex situ conservation is proposed.
Some of the populations grow into protected areas and
NATURA 2000 sites. The Limonium species, as an output of this work, have to be considered as efficient bioindicators for the conservation status of the coastal zone, while population dynamic is a reliable test for the conservation actions at local level. Moreover, the methodology applied to these three species could be extended to other Limonium species and used as a casestudy for microspecies ecology and chorology investigation in order to be implemented into rapid biodiversity assessment procedures.
P0031 – ePoster
Studies on diversity of coastal sand dune vegetation and stress biochemistry at coastal zones of Bay of
Bengal in India with special reference to its conservation
Chakraborty, T 1 , Mondal, AK 1 , Mondal (Parui), S 2
369

1 Vidyasagar University, West Bengal, India; 2 Lady
Brabourne College (under Calcutta University), West
Bengal, India
Coastal sand dunes are the important sites for a variety of flora, fauna and microbial elements. Coastal plant species have value-added nutritional, medicinal and agricultural uses along with their capacity to check the soil erosion and wind velocity, performing as a guard wall for the coastal habitat. Various natural forces and human interferences cause destabilization of coastal sand dunes and dangerously influence the dune ecosystem. Plant species adapted to coastal habitat are immensely useful in dune stabilization and habitat restoration. The research on dynamics of coastal ecosystems, threats and their conservation strategies are very limited. Our present study enlightens the ecology, floristic study and some biochemical investigation related to the stress-tolerance of the sand dune vegetation at the coastal zones of Bay-of
Bengal in India. We have detected several amino acids by thin layer chromatography among these proline showed the largest increase in the sand dune areas. The remarkable increases of soluble sugars are also noted.
P0032 – ePoster
Reintroduction and restoration of
Hubbardia
Bor, a monotypic, critically endangered grass genus in the
Western Ghats, India
Chandore, A 1
Gurav, R 1
, Gund, S
, Yadav, S 1
1 , NImbalkar, M 1 , Kedage, V 1 ,
1 Shivaji University, Kolhapur, India
Hubbardia Bor, a monotypic critically endangered and endemic genus in the Western Ghats was first collected by Sedgewick from Jog fall in 1919. It was described in
1950 by Bor. Hubbardia heptaneuron Bor considered to be endemic to Karnataka state, was presumed to be extinct from type locality as it could not be recollected from this place over a span of 8 decades even after intensive searching (Mishra & Singh, 2001). However, it was recollected for the first time from Tillari Ghat
(15˚48’083”N and 73˚ 10’042”E), a location other than type locality in Kolhapur district of Maharashtra in 2000.
Initially the grass was located in ghat region along road side on about 1 sq. meter, wet rock in shady place at c.
500 m altitude. Intensive search in consequent years in other ghat areas as well as its type locality could not locate the grass species. In year 2006 however, it was found growing in some nearby locations in Tillari region.
Taking into account the botanical importance, the unique habitat requirement and rarity of the species, a species recovery programme supported by Department of
Biotechnology (DBT), New Delhi, was initiated in 2006 by Shivaji University on restoration of the species in collaboration with ATREE, Bangalore and Forest
Department, Pune. Careful observations were made on phenology, seed setting, seed dispersal, seed germination and seedling establishment both in the field as well as in laboratory. Based on these observations, a programme was designed for reintroduction and restoration of the species. Conventional method of seed propagation was found to be the best suited for restoration of the species.
The species was reintroduced in 20 ghat regions at c. 120 locations in suitable habitats on vertical rock surfaces,
370 covering a stretch of 677 kms (air distance) from Jogfalls in south to Malshej Ghat in the north. The seeds germinated and established self-perpetuating populations in localities of reintroduction. The populations are monitored for their performance for the last 4 years.
Restoration of H. heptaneuron is successful in different localities of Western Ghats. Over 10,000 individuals at various locations have been established so far in Western
Ghats. This perhaps, is the original story in India of successful reintroduction and restoration of critically endangered species in Western Ghats involving efforts of
Department of Biotechnology (DBT), New Delhi; Shivaji
University, Kolhapur; ATREE, Bangalore and Forest
Department, Pune. The present paper deals with successful reintroduction and restoration of botanically important and globally threatened H. heptaneuron in
Western Ghats.
P0033 – ePoster
2
1
Endangered plants in Fagaceae from China and the reintroduction of
Quercus xichouensis
Chen, W 1 , Zhou, Z 2
Kunming
, Xia, K
Institute
1 of Botany,CAS, China;
Xishuangbannan Tropical Botanical garden, CAS,
China
The plants of Fagaceae are widely distributed from tropical to temperate forests and play a very important role in the Northern Hemisphere ecosystem. Many
Fagaceae members are dominant elements in forests.
However, several species of Fagaceae are endangered.
Nine genera and 900 to 1000 species are recorded in
China totally, 49 species grouped into nine genera on the
Red List of IUCN, about 5% Fagaceae species are under endangered status. A few of them are very rare based on the IUCN standard. Quercus sichouensis (Sichou Oak) is a most distinguished example. For this species only a total of five individuals survive in the wild. There are no remaining individuals in the type locality, even though the species was common in south-east Yunnan in 1947.
Even though the five individuals of Sichou Oak are growing and produce a lot of nuts every year, it is facing an extremely high risk of extinction in the wild. For rescuing Sichou Oak, the reintroduction and mechanism of endangered are carried on. The good news is there is no problem to germinate from this species. The optimal temperatures for germinating Q. sichourensis fruits were
25 and 30 ˚ C, of the germination about 88.0% and 84.0%.
89.5% seedlings could survive in the nursery garden.
There are 1000 seedlings growing in the nursery so far.
The 100 young trees are re-planted in its original distributed region and some other places in 2009 and
2010. To increase the individuals in the wild and also for the population regaining of Q. sichourensis , 35 seedlings have been replanted into Weishan county, about 800 km west from their original habitats in 2009. All new seedlings were growing well in the first year and 16 were left after extremely dry season in 2010. There are
100 seedlings are replanted in their original habitats in
2010. Based on the records, all replanted seedlings grow well. They grow 4.78 cm in height in average from them were replanted. The best one grows 11 cm. This critically endangered oak can be rescued with humans intervening.
The project is supported by National Natural Science

Foundation of China (3070056) and Yunnan provincial fund for applied basic researches and by the national fund for natural science (2008CD165).
P0034 – ePoster
Variation in plant communities and species composition along the microtopographic changes at the Lienhuachih Forest Dynamics Plot in Central
Taiwan
Chiu, S 2
Hsieh, C 6
, Chang, L 1 , Yang, K 3 , Wang, H 4 , Hwong, J 5 ,
1 Taiwan Forestry Research Institute, Taiwan; 2 Dept of
Botany, National Museum of Natural Science, Taiwan,
3 Dept of Ecology, Providence University, Taiwan;
4 Fushan Research Center, Taiwan Forestry Research
Institute; 5 Lienhuachih Research Center, Taiwan
Forestry Research Institute; 6 Institute of Ecology and
Evolutionary Biology, National Taiwan University,
Taiwan
The variations of the forest composition are primarily governed by topographic and climatic factors. How microtopography and microclimate affect the forest communities and species composition at one stand-level need direct and quantitative analyses. In this study, we investigated how the plant communities and species composition varied across fine-scale environmental heterogeneity at the Lienhuachih broad-leaved forest dynamics plot. All free-standing woody plants with diameter at breast height >1 cm of a 25 ha Lienhuachih
FDP were identified, measured, tagged and mapped at every 20*20m 2 subplot. Four plant communities were identified and represented with dominant and indicating species based on two-way indicator species analysis
(TWINSPAN). The Syzygium buxifolium – Pasania nantoensis type locating on the ridge and the highest elevation was with the highest stem density. The
Schefflera octophylla – Cryptocarya chinensis type locating on the upper slope was with the fewer stem density. The Cinnamomum subavenium – Cryptocarya chinensis – Tricalysia dubia type locating on the lower slope was with lower stem density but the highest species richness. And the Machilus japonica var. kusanoi –
Helicia formosana – Neolitsea konishii type locating on the valley or creak bank was with the lowest stem density. Further studies using Canonical correspondence analysis (CCA) were used which confirmed the results of
TWINSPAN. The first axis showed that convexity and elevation are the most important factors and the second axis showed that aspect is the important factor. Plant community and species at Lienhuachih FDP obviously differentiated in quantity and basal area relating to microtopography. It is possible that different microtopography influences the soil formation and so that affects the plant biomass and community types. It is also possible that, when disturbances occur, the reaction process and degrees from different microtopography are varied and result in the temporal variation in plant community and species distribution.
P0035 – ePoster
Regeneration of tropical
Acacia
species in response to fire
1
Congdon, B 1 , Williams, P 1 , Parsons, M 2
School of Marine & Tropical Biology, James Cook
University, QLD, Australia; 2 Queensland Parks and
Wildlife Service, Townsville, QLD, Australia
Most acacias are well known to regenerate prolifically from the soil seed bank following fire; however, some species have been observed to resprout from the base.
This study examines the germination and resprouting behaviour of 8 species – A. cincinnata , A. crassicarpa , A. flavescens , and A. mangium from tropical coastal woodlands and forests, and A. elachantha , A. hyaloneura , A. platycarpa , and A. ramiflora from the inland woodlands of White Mountains National Park in
North Queensland. Seeds of each species were subjected to dry heat at 40, 60, 80, 100 and 120°C and in water at
60 and 80°C for 5 minutes, and then incubated at 28°C.
Highest germination percentages were found for most species after treatment with 80 or 100°C dry heat or 80°C wet heat. Some 49% of seeds across treatments germinated over the first 140 days, whilst 10% of the remaining seeds germinated over a further 614 days, with one third of seeds remaining potentially viable after this time. In a pot study of resource allocation, eight individuals of A. cincinnata , A. crassicarpa , A.
flavescens , A. mangium , A. platycarpa and A. ramiflora ,
6 individuals of A. hyaloneura , and 3 A. elachantha were grown in 200 mm pots for 18 months. After harvesting the above-ground biomass, all of the wattles resprouted from the base, except A. hyaloneura and A. mangium . To further examine resprouting behaviour, nine 12 m x 12 m plots, separated by 4 m fire breaks, were established.
Acacias were planted at a spacing of 1.5 m x 3 m. Three replicates were allocated randomly to the three treatments – control, burnt and clipped. Seedlings were planted between October and December 2003, most by the end of November. Ten individuals of A. crassicarpa ,
A. flavescens , A. ramiflora and 8 of A. elachantha were planted in each plot, alternating between species. Fewer individuals of A. mangium (10), A. platycarpa (9), A. cincinnata (10) and A. hyaloneura (2) were available, and these were planted in even numbers across plots.
Three plots were burnt on August 2004. In 3 plots, the wattles were cut 3 cm from the base on September 2004.
Surviving plants were measured in September 2004 and
September 2005. Only one individual each of A. crassicarpa and A. mangium survived the fire treatment, while 3 individuals of A. crassicarpa , and one individual of A. ramiflora , A. elachantha and A. mangium survived the clipping treatment. Hence, survival was low, probably due to the competition from a high biomass of
Guinea Grass ( Megathyrsus maximus ), and the intensity of the fire fuelled by the high fuel load. The results indicate that germination of most species is favoured by a heat shock at 80°C for 5 minutes. All species but A. hyaloneura showed some ability to resprout, but few resprouted following an intense fire. For management purposes, low intensity fire may promote resprouting of many of these species, but high fuel loads, such as result from invasive grasses, will not favour resprouting.
P0036 – Poster
The real cost of carbon
Davidson, N 1,2,3 , Bailey, T 4 , Close, D 4
371

1 University of Tasmania, Australia;
Tasmania, Australia;
Australia; 4
3
2 Greening Australia,
CRC for Forestry, Tasmania,
TIAR, Tasmania, Australia
Carbon credits can be used to fund broad-scale restoration in the dry woodland landscapes of southern
Australia, once a price is placed on carbon in Australia.
The State Government of Tasmania (Climate Change
Office), Greening Australia and the University of
Tasmania have a $million project underway to demonstrate the practicality of this proposal. The ecological objective of the restoration project is to buffer and connect isolated remnants of native forest in the dry
Midlands of Tasmania in a way that will improve landscape scale connectivity in the face of climate change. Restoration plantings are designed to be biodiverse plantings of local native species (trees understorey, shrubs and ground cover) that reflect the forest structure and species composition of the 'original' forest thus optimising the biodiversity benefits of the restoration. To achieve the project objective we had to:
1. Determine the potential carbon storage in the dry forests in Tasmania (this was approximately 200t/ha C or
730t/ha CO
2 e)
2. Work with landholders and legal advisors to develop
130 year covenant agreements to secure our ownership of the Carbon in the trees while farmers owned the land
3. Choose land and agree on price with landholders
4. Adapt plantation forestry techniques to suit slow growing trees in a dry environment exposed to high levels of weed competition
5. At the three 30 ha sites chosen for the project, conduct replicated factorial experiments to identify: (1) the most suitable genotypes of the hardy local native eucalypts
(provenance trials involving 450 families of Eucalyptus pauciflora ); (2) the species mixture that would maximise productivity and carbon gain. Treatments were: single eucalypt species, mixed eucalypts, eucalypt and
Allocasuarina , eucalypt and Acacia , eucalypt and
Callitris , eucalypt and Bursaria , Allocasuarina monoculture, Acacia monoculture, grass and untreated control. Each of the species mixtures were planted with or without mixed species understorey. The real price of growing carbon that will also provide improvements in landscape scale biodiversity has to take into account of the pioneering work outlined in the points above.
However, once this technology is streamlined and commercialised we consider that carbon sequestration could provide an additional income stream to farmers on the low quality grazing country (steeper slopes and slower soils) and provide a new industry and employment in the dry landscapes of Australia.
P0037 – ePoster
Genetic conservation of
Ficus bonijesulapensis
R.M.Castro (Moraceae): landscape approach de Carvalho, D 1
F 2
, Fidelis Duarte, J 1 , de Almeida Vieira,
1 University Federal of Lavras, Brazil; 2 University
Federal of Rio Grande do Norte, Brazil
Ficus bonijesulapensis is endemic to deciduous forests on rocky outcrops and are distributed in disjunct areas of
Cerrado and Caatinga. Species of this genus are
372 considered key resources in tropical forests providing food during the periods of scarcity of other resources and, additionally, help in restoration of plant communities. Therefore, conservation of these species in its natural range while maintaining the structure of communities, contributes to the maintenance of the viability of populations in the long term and their genetic diversity. This diversity is directly linked to the evolving capacities of the species response to environmental changes. The identification of genetic discontinuities between populations performed by Delaunay triangulation helps to detect barriers that limit the gene flow among populations and the existence or not of spatial patterns in order to delineate management units
(MU) of genetic diversity. We used ISSR primers to analyze the diversity, structure, spatial patterns of genetic variability and discontinuity of 15 populations of F. bonijesulapensis . The genetic diversity (He) was 0.37 and the percentage of polymorphic loci was 100%.The
AMOVA showed that most of the diversity is within populations (77.18%) as expected for outcrossing plants and perennials. This may reflect the low gene flow (1.0), which is probably limited due to the isolation of outcrops where F. bonijesulapensis occurs, added to the different surroundings landscapes. The genetic structure inferred by Bayesian analysis showed that the genotypes of all sampled individuals can be divided into six groups.
However, there is a spatial pattern of genetic variability according to multivariate correlogram profile and that was confirmed by the Mantel test correlating the genetic distances (
θ β
) and geography (r=0.06, P=0.68). The analysis indicated that genetic disruption of the barriers among populations could be mainly mountain ranges and rivers. Six management units (MU) were proposed according to the location of genetic discontinuities, the values of diversity of populations and their geographical location. The aim was to enhance the MU's ability to maintain minimum viable populations and greater genetic variability.
P0038 – ePoster
Photoperiodic induction of synchrony in
Annona crassiflora
Mart. in the Cerrado southeast Brazil
De Carvalho, D 1 , Leao Bandeira de Melo, D 1
1 University Federal of Lavras, Brazil
Phenological behaviour among tropical trees has been a long-standing question for ecologists. Plant phenology in
Cerrado trees seems not be driver by single cue and appears to be lead by biotic and abiotic interaction over a phylogenetic heritage. To interrelate the whole complexity involved in the phenological behaviour of tropical trees seems to be necessary to study deep single species, trying to explain it whole fitness behaviour. The main objective of this study is to describe part of the fitness behaviour of adult trees of A. crassiflora , an important component of Cerrado canopy and belongs to a genus that is both ecologically important and commercially valuable. We recorded environmental cues, intensity and synchrony of phenological events of, leafing, flowering and fruiting, and interrelated them using Spearman’s correlation. A. crassiflora is a brevideciduous species. Rainfall is the main resource to support vegetative growth and day length play an

important role in vegetative bud break, working as a trigger for leaf flush. The species show a bimodal flowering behaviour during the year and anthesis synchrony only show correlation with photoperiodic variation, coinciding with the proximity to the equinoxes.
This phenomenon could be explained by the center of origin of the Annonaceae and the selective force toward to maintain the population of pollinator and guarantee the pollination service. A. crassiflora is dispersed by dung beetles, dispersing its fruit during the wet season, coinciding with dung beetle activity. It appears that in A. crassiflora reproduction is designed to suit the most favourable conditions for fruiting and seed dispersion rather than for flowering.
P0040 – ePoster
Phenology, water status and radial growth in two ecotypes of a tropical tree from the Brazilian Atlantic
Forest and Cerrado (savanna)
Lemos-Filho, JP 1
Paiva, EAS 1
, Toledo, MM 1 , Cândido, MVO.
1 ,
1 Departamento de Botânica, Universidade Federal de
Minas Gerais, Belo Horizonte, MG, Brazil
Understanding tree growth is essential to predict the effects of climate and human changes on vegetation structure and composition. Distinct ecotypes of
Plathymenia reticulata Benth. (Leguminosae,
Mimosoideae) occur in the Brazilian Atlantic forest and
Cerrado (savanna). The aim of this study was to compare the radial increment between ecotypes in a forestsavanna boundary, and to investigate the relationship of radial growth with climate, phenology and water potential. Band dendrometers were installed in fourteen individuals of each ecotype and the phenology and the water potential at predawn and midday were evaluated monthly. Mean comparison tests and Spearman correlation analyses were carried out with the data. Both ecotypes showed a positive radial increment during the months of high precipitation, but no increment, or decrement, in months of low rainfall. The radial variation was greater in the forest ecotype. Leaf-fall coincided with the cessation of radial increment. The Cerrado ecotype showed lower water potentials during the short drought period on summer, but during the winter dry season there was no significant daily variation. Radial growth was positively correlated with precipitation and temperature and negatively with deciduousness. The radial variation was due to cambial activity (growth) and tissue water status. The secondary tissues characteristics of the forest ecotype may have caused the greatest radial variation and a smaller drop in water potential during high evaporative demand conditions. Leaf shed coincided with cambial dormancy and may decrease water stress during the dry season. Precipitation was determinant in the phenology and growth of this species. Theses differences between ecotypes probably reflect in current and future distribution of P. reticulata . This study was supported by FAPEMIG (Fundação de Amparo à
Pesquisa do Estado de Minas Gerais) and CNPq
(Conselho Nacional de Desenvolvimento Científico e
Tecnológico).
P0042 – ePoster
The sexual polymorphism in a trioecious population of
Opuntia robusta
can increase the ecological versatility of this population in a human-altered
1 ecosystem
Del Castillo, RF 1 , Trujillo-Argueta, S 1
Instituto Politecnico Nacional CIIDIR IPN Oaxaca,
Mexico
The study of sexual polymorphisms is essential for understanding the evolution of sexuality and breeding systems. The presentation of both functional sexes in a single individual in contrast to a single functioning sex is one of the most extreme differences of the sex systems.
Most studies of the consequences of sex systems in plants usually are performed by comparing related species of different sex systems living in different kinds of habitat. With this procedure, it is difficult to disentangle the effects of the sex system from those of the habitat or the genetic background. In contrast, studying species with trioecious populations allows comparing the performance of male, female and hermaphroditic plants of the same species and in the same population. Opuntia robusta (Cactaceae) is one example of species with this kind of populations. A previous study conducted in Santa Rosa, San Luis Potosí, in central Mexico, shows that males and females potentially outperform hermaphrodites in the quantity
(males) and quality of their gametes. However, pollen limitation reduces greatly the reproductive potential of unisexuals to the point that they probably cannot compensate their full sterility of the non functional sex.
Hermaphrodites, on the other hand, can produce seeds autonomously with no evidence of inbreeding depression, reducing, in this way, interspecific pollination, and contributing to the pollen pool, as pollen discounting is negligible. Therefore, the maintenance of unisexuals in O. robusta is puzzling (del Castillo and
Trujillo 2009). In this study, we compared the vegetative performance of males, females and hermaphrodites in the same population. In particular, we monitored the growth rate of cladodes (the units of growth in Opunita ), the capacity of sprouting of manually detached cladodes, and the rates of water loss. We found that unisexuals are superior in growth rate, sprouting capacity and display lower transpiration rates than hermaphrodites. A faster growing rate of the cladode may also reduce the time at which the cladodes are tender and with soft spines, a stage with the highest vulnerability to herbivory.
Compared to other species of Opuntia , in O. robusta cladodes cannot be detached unless they are cut off manually, suggesting that the superiority in asexual reproduction in unisexuals is associated to human activities. To test this hypothesis we study the spatial distribution of unisexuals and hermaphrodites in the study population, and found evidence that unisexuality is mostly associated to places subjected to strong habitat alteration by humans, such as in the proximity of trails and fences. Thus, the maintenance of the sexual polymorphism in O. robusta can be explained by the contrasting advantages which exhibit the different sexual morphs. While hermaphrodites are superior in their sexual reproductive output, unisexuals are superior in their vegetative performance. This study shows evidence
373

of the role of anthropogenic activities in maintaining the sexual polymorphism in O. robusta . We propose that the diversity in sexual morphs increases the ecologic versatility of this species and can help explain the wide geographic and ecological distribution of this species.
P0045 – ePoster
Seed volume as an alternative to seed mass in ecological and functional studies
Dias, LS 1 , Ganhão, E 1
1 Dep. Biologia, Univ. Évora, Portugal
Seed size, a trait known to have a high ecological and functional relevance, is overwhelmingly expressed as average mass. However, to integrate ecological and functional correlates of seed size distribution in seedbank studies, seeds must be sorted by size which for practical reasons has to be done by sieving. Separation by sieves relies on linear dimensions of seeds from which volume can be easily calculated using published equations and implies that seeds quantification relate to intervals of linear dimensions. Data of length, width and thickness of individual seeds was assembled from a variety of sources, mostly from Iberian Peninsula, USA and Brazil
(50%, 30% and 12% of records) generating a seed volume inventory comprising by now 2693 species from
138 families. Two or more values of seed volume could be calculated for 75% of species. It has been predicted that plants with many resources have more variable seeds than plants with few resources and that the minimum seed size is independent of the amount of resources.
Therefore, it can be expected that within species, seed volume amplitude is better explained by the maximum than by the minimum volume. This hypothesis is supported by our data, with the minimum and maximum volume explaining 80% and 96% of seed volume amplitude respectively. Therefore minimum volume was chosen to investigate seed size ecological and functional correlates given its greater independence from environmental conditions during seed production. We investigated how seed volume compares with seed mass to explain light requirement for germination, expressed as relative light germination (RLG). Published data show that average seed mass explains 28% of average RLG.
The minimum seed volume can explain 20%, a lower but still meaningful fraction of RLG variation. The relationship between size and chemical defences of seeds and between size and nutritional value of seeds was investigated using published data of dihydroxyphenol, cyanoglycoside and soluble protein mean concentration in seeds. No significant relationship could be found between seed volume and cyanoglycoside concentration but seed volume could explain 22% of dihydroxyphenol concentration and 8% of soluble protein concentration, a value that rises to 37% if three clearly outlier species are removed from the analysis. However preliminary, our results suggest that seed size expressed by the minimum volume can be an adequate substitute to average seed mass in ecological and functional studies, making possible the re-examination of a large mass of published data and the integration of seed size distribution in seedbank studies.
374
P0046 – ePoster
Wood decaying fungi in
Abies cilicica
forest
1
Dogan, HH 1
Selcuk University, Turkey
In this work an attempt has been made to establish the qualitative composition of macromycetes, parasites and saprobes in fir ( Abies cilicica subsp. cilicica and subsp. isaurica ) distributed in Turkey. Turkey is rather insufficiently studied from a mycological standpoint, and the number of recorded macromycete species is much smaller compared to some other Mediterranean countries.
Research has been conducted in more than 30 localities in the central and southern parts of the country between
1,400 and 1,800 m altitude. Research has been done in most of the localities where its associations are best developed. The majority of these localities are in Konya,
Adana, Mersin, Antalya districts, on the slopes of Taurus
Mountain. A total of 150 macromycetes species belonging to Basidiomycota and Ascomycota have been identified. Amaurodon , Cabalodontia , Ceraceomyces ,
Craterocolla , Crustoderma , Ditiola , Fibulomyces ,
Leptosporomyces , Peniophorella , Resinicium ,
Trichophaea and Veluticeps genera were found for the first time in Turkey. The distributions according to the ecological status of the species are as follows: parasites
(50), lignicolous (100). Many of the macromycete species in fir forest are very rare, threatened or with small disjunct ranges. Some of them are indicators of specific, endemic habitat types or are interesting for other reasons. The importance of the species and their ecologic position in the fir forest are discussed and some brief knowledge are also given for new and rare species.
P0047 – ePoster
Phosphorus conservation constrains tree responses to rising atmospheric [CO
2
] in native Eucalyptus woodland
Ellsworth, D 1 , Crous, K 2
1 University of Western Sydney, Australia;
National University, Canberra, Australia
2 Australian
Global atmospheric CO
2
concentrations are rising at a rate exceeding nearly 2% per annum and as [CO
2
] is enriched, a stimulation of leaf net photosynthesis and the ecosystem carbon cycle is expected. Given the vast global extent of native forests, stimulation of photosynthesis and tree growth as well as C storage by elevated [CO ] has large potential offsetting effects on
2 further increases in atmospheric [CO
2
]. However, nutrient limitations can pose strong constraints to such growth and C storage increases as has been documented for nitrogen-limited temperate forests. While the effects of elevated atmospheric [CO
2
] on ecosystem carbon cycles are well-documented for forests on relatively young and fertile soils, little work has been conducted on ancient soils with limited nutrients and limited P supplies for growth. We describe a free-air CO
2
enrichment experiment ‘EucFACE’ being implemented in a mature native Eucalyptus woodland in western Sydney,
Australia, the first-ever ecosystem FACE study in old-

growth forest vegetation. The study forest occurs on heavily-weathered, ancient soils with low P supply to plants. Phosphorus limitations to plant production predominate throughout Australia’s landscape and globally on ancient soils in tropical and subtropical regions, and the nature of these limitations are poorly studied in most regions of the world. We present theory and evidence that photosynthesis of native sclerophyll trees growing on P-poor soils is constrained by carbohydrate export capacity, particularly at high intercellular [CO
2
] levels, corresponding to 500–600 ppm atmospheric [CO
2
]. This sink limitation and carbohydrate export hypothesis, first proposed over 100 years ago and utilised in practical cultivation of fruit trees, is tested by a series of experiments on native plants within
‘EucFACE’. Whilst the short-term mechanism of photosynthetic response to elevated [CO
2
] would suggest a large CO
2
stimulation effect in forests, the longer-term response is very different from this. The findings have relevance to explaining responses of evergreen broadleaved woodlands to atmospheric change, and provide clues to how the significant fraction of global ecosystems that are phosphorus- and water-limited may alter functioning in a future, higher CO
2 anticipated in the next 40 years.
atmosphere as is
P0048 – ePoster
Diversity and evolution of Araucariaceae: in pursuit of ghost lineages from South American fossils
Escapa, I 1 , Cúneo, R 1 , Wilf, P 2
1 Museo Paleontológico Egidio Feruglio, Patagonia,
Argentina; 2 Pennsylvania State University, USA
In South America, the family Araucariaceae is represented by two living species of the genus Araucaria section Araucaria , A. araucana and A. angustifolia , which along with the Australasian species of Araucaria ,
Agathis , and Wollemi define a mostly Southern
Hemisphere distribution for the family. In addition, the
Araucariaceae shows an extended fossil record with occurrences recognized since the early Mesozoic in both hemispheres. Stratigraphic adjustment of phylogenetic hypotheses for the family indicates that divergences leading to the major clades probably occurred as early as the Early Jurassic. Despite the extensive fossil record of the family, the phylogeny indicates ghost lineages for several genera and sections. We discuss the rich record of the family in South America and its implications for understanding the evolutionary relationships and the geographic distributions of past and present species.
Several of these taxa fill remarkable temporal and morphological gaps in the record of Araucariaceae and therefore provide a more complete assessment of the evolutionary history of the group. New findings from
Jurassic, Cretaceous and Paleogene localities in
Argentina and Colombia are highlighted in this evolutionary context.
P0049 – ePoster
Intraspecific competition in initial stages of the moss
Tortula muralis
: spore-density dependence of germination and protonemal development
Estebanez Perez, B 1
N 1
Nunez Martin, C 1 , Garcia Medina,
1 Universidad Autonoma de Madrid, Madrid, Spain
Little study has been made on intraspecific competition in mosses, especially in initial developmental stages.
Here we present an in vitro test for density-dependence in spore germination and protonemal development in the moss Tortula muralis , one of the most abundant and widely distributed species in the Mediterranean region.
We have analyzed the dynamics of germination and protonemata development through a series of experiments using widely different spore densities
(ranging from 1,600 to 240,000 spores/ml), in standard conditions for in vitro liquid culture of bryophytes: medium was half-strength Murashige & Skoog inorganic formulation (MS1/2), at pH 7, and culture chamber conditions were 25ºC and 16 h light / 8 h dark photoperiod. Spore germination rate and protonematal development were observed and recorded every 5–7 days for five weeks. Our results show a clear effect of spore density on germination and, most especially, on cell division. The germination rate is high (over 90%) in all cases; however, it is significantly delayed for spore densities over 16,000 spores/ml. In turn, the development of multicellular stages (unbranched and branched protonemata) was observed as clearly impaired from the first week in dishes with even lower spore densities (over
4000 spores/ml). A sigmoidal dose-inhibition curve was fitted by nonlinear regression for density vs. development
(R2 >0.90). A series of parallel experiments varying the nutrient concentration in the culture media show that the inhibitory effect of spore density was not reverted using higher nutrient concentrations (MS1 or even MS2).
Besides, we observed the ability of the spores in this moss species to germinate and produce protonemata in very low mineral content (MS1/40). This suggests that competition for nutrients is not the only factor mediating these intra-specific interactions, and points to the possible production of an inhibitory chemical factor. The inhibition effect of the spore density on initial development stages must be taken into account in moss culturing, including ex situ conservation programs, and could play a major role in the establishment of natural moss populations.
P0050 – ePoster
Comparative analysis of lead tolerance in urban and non-urban moss species
1
Estebanez Perez, B 1
Medina, N 1
, Menendez Martinez, M 1 , Garcia
Universidad Autonoma de Madrid, Madrid, Spain
We present here a comparative study of the toxic effects of lead on germination and protonemal development in urban and rural mosses. We have selected three families of mosses, and within each family, a pair of related species, one of them characteristic of urban environments and the other typical of unpolluted sites. The taxa selected list as follows: Fam. Pottiaceae, Tortula muralis
(urban) and Tortula canescens (rural); Fam.
Grimmiaceae, Grimmia pulvinata (urban) and G. orbicularis (rural); and Fam. Orthotrichaceae:
375

Orthotrichum diaphanum (urban) and Orthotrichum tenellum (rural). A sequence of increasing lead acetate concentrations (ranging from 0 to 10 millimolar) was tested on mature spores grown in half-strength
Murashige & Skoog inorganic liquid medium, pH adjusted to 6 or 7. Spore germination and protonemal developmental stages were monitored for 28 days. In all cases, response to lead follows a typical sigmoidal model of log transformed dose vs.response model both for germination and for cell division. In the Pottiaceae and
Grimmiaceae, but not in the Orthotrichaceae, the lethal dose and the cell division inhibitory dose (ID50) were higher in the urban moss representative than in their rural partner. Different degrees of tolerance were detected between families. In Tortula muralis (Pottiaceae), even using 100 micromolar lead concentrations, both branched and unbranched protonemata were produced in an abundance comparable to that observed in the control dishes, and spores were shown to germinate in lead concentrations of 1 millimolar; thus showing higher lead tolerance than previously reported for mosses. In contrast, the spore germination and protonemal development in the urban member of Grimmiaceae was observed as much more sensitive than expected. Lead tolerance in initial stages is pointed out as a major selective constraint in urban environments at least for some mosses, although in other species, such as the
Orthotrichum pair analyzed here, their different urban tolerance must be conditioned by their diverse sensitivity to other pollutants or stress factors.
P0051 – ePoster
Spore ultrastructure and viability in moss species of the genus
Orthotrichum
with different dispersal strategy
1
Estebanez Perez, B
Callejo, R 2
Atauri, I 1
Garcia, F 1
1 , Garcia Medina, N 1
, Medina Bujalance, R
, Ruiz Herranz, A 1
, Caparros
, Garilleti Alvarez, R
, Mazimpaka Nibarere, V 1
1 , Draper y Diaz de
2 , Lara
Dpto Biologia, Universidad Autonoma de Madrid,
Spain; 2 Dpto Botanica, Fac. Farmacia, Universidad de
Valencia, Spain
The different modalities of spore dispersal in mosses are considered as a crucial factor in the establishing strategies of populations and the dynamics of the distribution areas of each particular species. Different biological aspects may, in turn, affect the competence of the spores to survive dispersal and establish a new colony. We have undertaken an ultrastructural and viability study on 6 moss species belonging to the genus
Orthotrichum sect. Gymnoporus , where two different spore dispersal strategies are represented: a) xerocastique dispersal (spores released in dry conditions), and b) hygrocastique dispersal (spores released in wet conditions). Electron microscopy (TEM and SEM), as well as germination tests of spores subjected to stress
(heat and prolonged storage), have been performed. Our observations show no clear-cut differences between hygrocastique and xerocastique species regarding spore wall thickness, starch and lipid storage, and plastid development, although there are some examples of endosporic germination restricted to hygrocastique representatives. However, viability of spores in
376 hygrocastique species shows a greater decrease when subjected to heat and storage stress. According to our results, the diverse sensitivity of the spores to stress conditions could explain to a certain extent the different dispersal strategies, although some other factors affecting the survival success in other cycle phases must be involved.
P0053 – ePoster
Agriculture intensification through irrigation results in a decline of arable plants diversity in a
Mediterranean area
1
Fagundez, Jaime 1
Gómez, D 1
, Tejedo, P 1 , Mateo, P 1
, Pérez-Olea, P 1
IE University, Segovia, Spain; 2
, Castro, V 2 ,
University of León,
Spain
Agriculture intensification is a major threat to biodiversity held in agroecosystems, and arable plant species are one of the groups more directly affected by these changes. In Castilla y León (Central Spain), intensification is rapidly taking place mainly by means of the transformation of extensive cereal dryland to irrigated farmland This results in deep changes over management practices such as crop rotations or nitrogen inputs.
Effects of these major changes on the environment, and particularly on biodiversity, have been very little studied despite of the extensive areas to be affected in years to come. Here we study the effects of agronomical intensification through irrigation on arable plants richness, diversity, functional traits and community composition. The study was carried out in an extensive area where both traditional extensive dryland and irrigated farmland areas occur. 80 fields were selected,
40 dryland and 40 irrigated land. The only crop in dryland is wheat. Irrigated fields where divided in three categories according to crop (wheat or maize) and irrigation (drip or sprinkle-irrigation). Irrigated fields had significative lower values for total species richness, mean species number per field and beta diversity. Floristic composition is also affected, with higher percentages in irrigated fields of perennials vs annuals, cosmopolitan and exotic vs native, and wind-pollinated vs insectpollinated. Highest differences are found for both types of maize fields where many species are exotic and/or C ,
4 and some represent a severe pest such as Cynodon dactylon . We discuss that future transformation of large areas to irrigated farmland will represent a strongly negative effect on biodiversity, and that environmental costs of this intensification should be taken into account on agronomy policies.
P0054 – ePoster
The study on the interactions between the branching of arbuscular mycorrhizal fungi and parasitic plant seed germination by using strigolactones as hastening stimulus
Feng, D 1 , Chen, GL 1
1 Inner Mongolia University,Huhhot, PR China

Strigolactones (SLs) is the generic name of a collection of organically related compounds which are derived from carotenoids and synthesized in roots of most land plants including the monocots: Sorghum , maize and proso millet, and the dicots: cotton, cowpea and red clover.
Originally, Strigolactones have two closely related functions. One is triggering seed germination of the parasitic plants as principal plant-derived signals; the other is promoting arbuscular mycorrhizal (AM) fungi branching and symbiosis. While their newly discovered function is to act as hormonal signals to inhibit bud outgrowth and shoot branching, together with auxin and cytokinin. These discoveries explain the natural role of strigolactones during plants evolution. Scientists have paid due attention for a long period of time to strigolactones which act as a stimulus to parasite seed germination. Parasites such as Striga and Orobanche , which parasitize plant roots and rob water and nutrients of hosts, are considered to be the scourge of agriculture in many parts of the world. A single mature Striga plant is able to produce thousands of seeds, which may stay dormant in the soil for up to 20 years until they sense the growing roots of a potential host and then begin their germination. Germination of the parasite seeds is initiated by recognizing certain chemical signals secreted by the roots of the host plant. Then the germinated seeds grow toward a host until they attach the host plant’s root via a haustorium, through which the parasite seedling draws nutrients from the host. Strigolactones are also known as the branching factor of stimulating AM fungi to branch in order to penetrate and colonize plant roots, receiving carbohydrates derived from photosynthesis in the host and providing essential nutrients such as phosphate, nitrate and other minerals from the soil to the host. Hyphal branching is the original step in host recognition by AM fungi. Host roots release branching factors to induce extensive hyphal branching in AM fungi which produces Myc factors to induce the molecular and cellular responses leading to successful root colonization by AM fungi. Therefore, strigolactones are accepted as the branching factors. There is another speculation that the direct germination effect on parasitic plant is not owing to the strigolactones but due to the AM fungi branching and the symbiosis with its hosts which strigolactones promoting. This encourages us to investigate the interactions between AM fungi branching and parasitic plants seeds germination hastened by strigolactones, especially the germination of two kinds of parasitic plants, Cynomorium songaricum and Cistanche deserticola , both of which are traditional Chinese medicine. These researches may provide some useful information for the study of mechanisms of parasitic plants germination as well as its related field.
P0055 – ePoster
Treasure in the junk: domesticated transposable elements and low temperature tolerance
, Bureau, T 1
1
Forczek, E 1
McGill University, Québec, Canada
Transposable elements (TEs) are independently replicating sequences that can move in the host genome.
Because of this ability, TEs were labeled as junk, selfish or parasitic elements, until recently. In the last two decades, a growing number of studies have documented cases where TEs have lost their mobile lifestyle and become part of the host gene repertoire by a process called molecular domestication. However, the biological function of most of them is still unknown. One example of domesticated TEs in plants is the MUSTANG (MUG) gene family, which was originally uncovered computationally. MUG was derived from the superfamily of Mutator-like elements (MULEs) and is present in a majority of angiosperms, but not among gymnosperms.
This might suggest that MUG was domesticated just as flowering plants began their rapid evolutionary diversification. In Arabidopsis , the MUG gene family consists of eight members divided into two subfamilies, based on their conserved domains. Subfamily A (MUG1-
MUG4) consists of three domains, MuDR, MULE and a
SWIM (subfamily A) and Subfamily B (MUG5-MUG8) has an additional PB1 domain. In this study, we focus on two genes, MUG7 and MUG8. Exploiting T-DNA insertion mutants, we identified and selected four homozygous mutants lines for MUG7 and two for
MUG8 and crossed them to generate double-mutant, mug7mug8, lines. Phenotypic analyses including developmental timing, inflorescence height, flowering time and seed set indicate that each single mutant has a specific phenotype and double mutants have the most severe phenotype, a dwarf form. We also subjected mug7 single mutants to a cold stress assay. Plants were kept in either at +13°C or +4 °C from stratification until the end of their life cycle. Phenotypes were much more pronounced than under normal growth conditions
(+22°C); however, these cold stress conditions had no obvious effect on fitness. Furthermore, when subjected to a freezing tolerance experiment, one of the mug7 alleles has freezing tolerance for temperatures as low as -12°C when compared to wildtype and mug8 mutants. At higher temperatures from 0°C to -4°C, most of mug7 mutants can survive without any serious damage. Among domesticated MULEs, there are genes whose functional role has been experimentally confirmed. FAR1 and
FHY3 are transcription factors responsible for modulation of phytochrome A signaling pathway and effect plant light response. FAR1 and FHY3, except for the PB1 domain, have similar conserved domains, what might suggest that MUG7 could also be a transcription factor. The difference however is in the functional outcome and highlights the important roles that domesticated TEs might play in host evolution.
Furthermore, domesticated TEs are virtually an untapped resource that may underlie many agronomically important traits and should be aggressively examined.
Specifically, we hope our study might contribute to the improvement of agronomically important Brassicaceae species, such as canola, by exploiting the unique freezing tolerance feature of MUG7.
P0057 – ePoster
Climate change and its implications on the biodiversity of the biosphere reserve 'Pantanos de
Centla', Tabasco, Mexico
Gama, L 1 , Collado-Torres, R
Díaz-Lopez, H 1 , Moguel-Ordoñez, E 1
Pacheco-Figueroa, C 1
1 , Villanueva-Garcia, C 1
, Ortiz-Perez, M 1
, Valdez-Leal, J 1
,
,
1 Universidad Juarez Autonoma de Tabasco, Mexico
377

Tabasco is located on the southeastern part of Mexico on the Grijalva-Usumacinta basin with the biggest and most dynamic rivers of the region. One third of the all fresh water reservoirs of the country pass through it. The
Biosphere Reserve Centla Swamps was created on 1992 and covers 302,706 hectares, represents the 12% of
Tabasco's area and 11% of the total swamps within
Mexico. The biosphere contains almost 600 different species of plants. Due to its location, it´s one of the most vulnerable areas to climate change impacts in Mexico.
And evaluation of the potential conservation of the area was require to promote protection policies related to the effects of climate change especially to environmental services that will help propose strategies for conservation. The objective was to identify the degree of conservation of the system, as well as identifying threats that could increase the impacts of climate change. A geographical information system was use to configure a cartographic data base of actual and historical maps to find out changes on it. A review of a set of landscape components was done to relate them to natural changes on the hydrodynamic and their probable impacts on the territory. Maps with different landscape components as well as climate change scenarios were use to identify vulnerable areas. Results show that current threats are floods, fires, and increase on salinity related to sea level elevation as well as and sea coast erosion. The baseline scenario shows that by the change in sea level would affect 75% of the reserve which will result on significant movements of species of flora and fauna. Because this is urgent to determine the species that is threatened and generate migration corridors to enable them to areas of refuge or so as to raise the location of new protected areas.
P0059 – ePoster
Phylogeography and genetic diverstiy of the endangered species
Taxus wallichiana
Gao, Lian-Ming 1 , Liu, Jie , Moller, 1 , Li, De-Zhu 1
1
Michael 2
Kunming Institute of Botany,Chinese Academy of
Sciences, China; 2 Royal Botanic Garden Edinburgh, UK
Abstract: The collision and post-collisional convergence of the Indian subcontinent with Eurasia resulted in the uplift of Qinghai-Tibetan Plateau (QTP) which dramatically changed the topography and climate of
Asia. Climatic oscillations in the Quaternary played a major role in species expansion and population structure.
In this study, the chloroplast DNA trn L-F region sequencing and nine microsatellite loci genotyping of
815 individuals from 43 natural populations of
Himalayan yew ( Taxus wallichiana Zucc.), an endangered tree species occurring in the East Himalayan and Hengduan Mountain (HM) were performed. Twentynine cpDNA haplotypes were detected in the sampled populations, which were divided into two lineages on phylogenetic and network analyses. The distribution pattern of the cpDNA haplotypes showed a strong phylogeographical structure (NST = 0.768, GST = 0.469,
P < 0.01). The divergence time of the two lineages dates back to 2.35–5.32 Ma, supporting the two lineages diverged by the rapid uplift of the QTP. The SSR data of the same samples well correspond with the cpDNA
378 results. Higher level of the genetic diversity was detected in EH populations than that in HM populations. The genetic diversity decreasing westward from China to
Nepal alone the Himalayas was found in EH populations.
Strong natural barrier was found between the two lineages. The mismatch distribution results revealed that the HM lineage may experience a seriously bottleneck effect in the Last Glacial Maximum (LGM), and then a recent rapid range /demographic expansion. And the East
Himalayan lineage probably survived in separate refugia and underwent several expansion/contractions during glacial/inter-glacial cycles in Quaternary.
P0060 – ePoster
Extension of winter down-regulation among different plant functional types
Garcia-Plazaola, JI 1
1
, Fernandez-Marin, B 1 , Esteban, R 1
1
Becerril, JM
University of Basque Country, Spain
,
In terrestrial ecosystems in which winter is enough cold as to inhibit photosynthesis, plants must face the risk of overexcitation of the photosynthetic apparatus generated by the unbalance between light absorption (which is temperature independent) and energy utilisation (which represents an enzymatic, temperature-dependent process). Under these conditions many plant species lose their leaves, while others remain green. One of the strategies that evergreen plants adopt during winter is to down-regulate photosynthetic efficiency maintaining a sustained thermal energy dissipation. This mechanism implies a decrease, and eventually the suppression, of photosynthesis, but allows the safe de-excitation of chlorophyll molecules within the antenna complexes.
Once conditions become favourable, photosynthetic structures need a certain time (typically several days) to be fully operative. So there is a trade off between the maximization of carbon assimilation and the achievement of effective photoprotection. This process, also called winter photoinhibition, can be easily detected by the presence of sustained depressions of Fv/Fm bellow the optimal value of 0.75-0.83 that are maintained even after several hours at warm temperature. This process of photoprotective photoinhibition has been deeply studied in boreal conifers, broadleaf evergreen trees and in some annual crops, but it is not known whether is a general phenomenon among photosynthetic organisms exposed to low temperatures or is an exclusive adaptation of some temperate plants. Thus, the aim of the present study is to study the extension, plasticity and relevance of this mechanism in different plant functional types naturally exposed to long periods of subfreezing temperatures.
Evergreen mosses, ferns, lycopods, gymnosperms and angiosperms belonging to three categories (herbaceous, shrubs, trees) were collected in the field and maintained
12 hours in the dark at 20°C to allow the recovery of photosynthesis. In these plants Fv/Fm was measured 30 minutes after collection and 12 after collection, allowing the estimation of the relative contribution of chronic and dynamic photoinhibition, considering 0.8 as the optimal value in non-stressed plants. Divergent strategies among functional groups were observed, but also among species coexisting in the same ecosystems. This study highlights

the importance of this mechanism as a key attribute for plant fitness.
P0061 – ePoster
The assessment of plant biodiversity on and off mature stands of
Androstachys johnsonii
Prain and
Colophospermum mopane
(J.Kirk ex Benth.)
J.Leonard
Georginah, M 1 , Ligavha-Mbelengwa, M 1 , Bhat, R 1
1 University of Venda, Limpopo Province, South Africa
The species that were growing on the stands of C. mopane and on open space stands weighed more than those on the stands of A. johnsonii in both loam and sandy soil areas. This is probably an indication that there are some toxic chemicals that are excreted from various parts of A. johnsonii that caused the suffering and hence lesser weights of the understory species of A. johnsonii .
The conditions were found to be unfavourable for the majority of the understory species to grow and flourish well under the canopies of A. johnsonii . This therefore, might imply that even though A.
johnsonii possess allelochemical materials, some understory species may adapt to such conditions in the proximity of its vicinity.
The decomposition of different parts of the plant on the ground surface under A. johnsonii canopies probably results in relatively highest concentration of allelochemical materials in the soil on A. johnsonii stands. Limiting factors like shading, temperature, water availability, and soil type may not necessarily be causes of the observed differences as the investigation was carried out from the same areas. The present investigation revealed that A. johnsonii in woodlands chokes its understorey plants and such understories might eventually die out.
P0062 – ePoster
Biodiversity information management tool for identification of plant conservation priority areas in
Tanzania
Gereau, RE 3 , Kindeketa, WJ 1
, Ward, H 4
, Stephen, SA 1 , Madadi,
1
LM 2
Tanzania Commission for Science and Technology, Dar
2 es Salaam, Tanzania;
Garden, St Louis, USA;
Tanzania Forestry Research
Institute, Lushoto, Tanzania; 3 Missouri Botanical
4 NetWings Company Inc., Boise,
ID, USA
In Tanzania, total plant diversity has not been considered as a major basis for the establishment of protected areas, in part because comprehensive and reliable information on plants was not previously available in a unified, readily accessible format and therefore could not be used to inform conservation planning and environmental decision-making. The documentation of the more than
10,000 higher plant species in Tanzania is widely scattered in numerous publications and herbarium specimens deposited in various institutions, most of which are not readily accessible to researchers and conservation practitioners working in Tanzania due to cost and logistical difficulties. The project will collate data on the vascular plant flora of Tanzania from sources both inside and outside the country together with relevant geospatial information into a comprehensive online biodiversity database, adopting the DarwinCode
Standards in use by the Global Biodiversity Information
Facility (GBIF) and the Tanzania Biodiversity
Information Facility (TanBIF) to facilitate data sharing.
The database will be used for assessment and identification of geographic and taxonomic gaps in currently available data and for delineation of areas of high plant conservation priority. The database will be the repository of all components of Tanzanian biodiversity, from which species checklists of all organisms can be obtained; conservation assessment of species based upon
IUCN Red List threat categories can be derived; and ecological, land use, and climate change modeling performed. The database will thus serve as a reliable source of information for environmental policy decisions.
P0063 – ePoster
Is genetic diversity analysis enough to approach conservation of the neotropical palm
Butia eriospatha
?
, Reis, MS 1
1
Nazareno, AG 1
Universidade Federal de Santa Catarina – UFSC,
Brazil
The long-living palm Butia eriospatha (Martius ex
Drude) Beccari (Arecaceae) is native to the Atlantic Rain
Forest, and grows in altitude grasslands. The current remaining populations of this vulnerable palm species consist of adult plants and seedlings, with no evidence of recruitment to later stages of maturity due the cattle farming. Furthermore, other adverse factors have threatened B.
eriospatha , such as loss of habitat due to exotic tree species’ reforestation, illegal sale of specimens in both local and international trade, and overexploitation of fruit. In order to estimate the current conservation status of B. eriospatha , we investigate the genetic diversity of adult plants and seedlings, the patterns of natural regeneration and the herbivory rate in one B. eriospatha population in Southern Brazil. In B. eriospatha population, all adult plants and seedlings were tagged with a numbered aluminum plate and mapped.
Nine microsatellite markers, previously developed for B. eriospatha , were used for genetic analyses. The B. eriospatha population studied consist of 44 adult plants and 29 seedlings, which show lower seedling recruitment and high rates of livestock herbivory (69%). The gene diversity was higher in seedlings (HE=0.504) than adult plants (HE=0.407) suggesting that inbred lines are intercrossed. Although the gene diversity seems to be preserved, we observed fixation of one allele resulting in eight polymorphic loci. Based on our observations, B. eriospatha may be able to reproduce by selfing
(geitonogamy), which may increase the homozigosity.
Contrary to expected for small populations, we observed a decrease of inbreeding effects in seedlings (f=0.171) as compared to adults (f=0.348). We hypothesized that the reduction of 50% of fixation index in seedlings can be due to the presence of recessive deleterious mutations reducing the survival of offspring as was, apparently, observed. Future studies about fixation index in B. eriospatha embryos can elucidate this question.
379

Furthermore, we did not detect unique alleles in seedlings indicating that this population, located 2 km apart from the next, can be isolated. This study highlights the importance of analysing both ecologic parameters and genetic data to better understand the threats facing a species. Clearly, our results suggest the urgent need for planning of the conservation and use of this biological resource. Monitoring and combating the illegal trade of adult plants, preventing the entry of cattle into the species occurrence areas, and facilitating the gene flow among populations are urgently required.
P0064 – ePoster
Genetic diversity among natural populations of
Butia eriospatha
, a vulnerable palm species from the
Atlantic Rainforest
Nazareno, AG 1 , Reis, MS 1
1 Universidade Federal de Santa Catarina – UFSC,
Brazil
The Brazilian palm flora is very rich, with estimates of about 400 species. Tropical palm species from the
Atlantic Rainforest, however, are experiencing reducion in population size due to current destruction and high deforestation rates. In spite of this, more research regarding the palm flora and its specific threats are necessary to generate useful information to plan conservation strategies. From a conservation perspective, we developed microsatellite markers in order to investigate and contribute for the knowledge about population structure and genetic diversity in remaining populations of the vulnerable palm species Butia eriospatha (Martius ex Drude) Beccari. Fourteen primer sets were isolated and characterizated in 90 reproductive individuals from three natural populations located in
Santa Catarina State, Southern Brazil. Genetic diversity parameters such as allelic richness (A), observed heterozygosity (HO), and expected heterozygosity (HE) were calculated. The levels of inbreeding were estimated using the equation f = (HE – HO)/HE. To test genetic differentiation between populations, we used the standardized G’ST Hedrick’s statistic. For the microsattelite set, five loci were monomorphic with alleles fixed in all B. eriospatha populations surveyed.
The number of alleles per locus ranged from 2 to 6; the observed and expected heterozygosities ranged from
0.265 to 0.460 and from 0.289 to 0.407, respectively. A total of 35 alleles were identified and the average number of alleles ranged from 2.25 to 3.11. Significant inbreeding effect (f=0.34) was observed just in the smallest population. Additionally, this population – with only 44 adult plants - contribute severally for the very high genetic differentiation among populations
(G’ST=0.56), especially due to a 65% deficit in heterozygotes observed in relation to the expected for
Hardy-Weinberg equilibrium. Nevertheless, because of the presence of rare and unique alleles in all populations, and high level of genetic differentiation, we emphasize the importance of the conservation of these populations, even highly fragmented.
380
P0065 – ePoster
Can extensive cross pollination buffer the effects of habitat fragmentation in tropical fig trees?
Nazareno, AG 1
Mestriner, MA 3
, Coelho, LFM
, Pereira, RAS 2
2 , Alzate-Marin, AL 3 ,
1 Universidade Federal de Santa Catarina – UFSC,
Brazil; 2 Faculdade de Filosofia, Ciencias e Letras de
Ribeirao Preto - FFCLRP/USP, Brazil; 3 Faculdade de
Medicina de Ribeirao Preto - FMRP/USP, Brazil
In population genetics a more general approach of landscape structure and processes on populations provides insight for understanding how species with different life history respond genetically to forest fragmentation. Nuclear microsatellite and landscape information were combined to answer how Ficus citrifolia P. Miller (hemy-epiphyte) and F. eximia Schott
(free-standing) respond to habitat loss at broaden spatial scale. We assessed three large plots of 100,000 ha representing three landscapes with different levels of forest fragmentation located in Semideciduous Atlantic
Forest fragments in Southest Brazil, Sao Paulo State.
Twelve microsatellite loci previously developed for
Ficus insipida were used to screen 24 to 82 individuals of F. citrifolia and F. eximia from each of the three studied populations. In all the populations analysed, we detected high levels of genetic variability and no inbreeding in both species. We detected loss of allelic richness in both fig species suggesting that intrinsic biology traits of species (e.g. long distance cross pollination) does not buffer the impacts of forest fragmentation. In spite of presence of rare and unique alleles, and high interpopulation genetic differentiation we emphasize the importance of conservation of these forest remnants. Our results contribute to better understand the dynamic of population genetics at broaden spatial scale, because the current knowledge about population genetics in Ficus is limited to intrapopulation surveys, with no inference on forest fragmentation effects.
P0066 – ePoster
Tough love: how drought in the maternal
1 environment affects seed dormancy
Gorecki, M 1 , Long, R 2 , Flematti, G 3
School of Plant Biology, University of Western
Australia, Crawley, Australia;
Garden, West Perth, Australia;
2
3
, Stevens, J 2
Kings Park and Botanic
School of Biomolecular,
Biomedical and Chemical Sciences, University of
Western Australia, Crawley, Australia
Seed dormancy delays germination in unfavourable conditions to facilitate seedling survival, and is an important ecological trait for plants in changing climates.
In weed species, seed dormancy complicates management, as sporadic germination allows plants to avoid being controlled. In native species used for restoring mined or degraded lands, seed dormancy can reduce the efficiency of the restoration effort. Despite these important implications of seed dormancy, little is understood about the factors that control it, and thus how

it might be affected by changing climates. On one hand, seed dormancy can be considered a polygenic trait with continuous phenotypic variation, but it can also be substantially influenced by the conditions experienced by the parent plant during seed development. The effect of particular maternal environmental conditions on the developing seed may depend on the level of stress perceived by the parent plant. Stress, such as occurring under drought, may be signalled in the plant by the hormone abscisic acid, which is also involved in maintaining seed dormancy. Therefore, variation in seed dormancy in a population may be due to genetic variation, or phenotypic plasticity in response to the maternal environment. This study aimed to explore the relative effects that maternal environment and genetics have in determining seed dormancy characteristics. Seeds from Brassica tournefortii , an invasive weed in southern
Australia, were collected from different locations in south-west Western Australia. We tested the effects of drought and population source on seed dormancy. Plants were grown either in a common garden, in the field, or in the glasshouse where they were subjected to drought stress, and the seeds were collected. To distinguish subtleties in dormancy state, we tested the germination response of seeds to the smoke-derived chemical karrikinolide (KAR1), which promotes the germination of B. tournefortii . The concentrations of abscisic acid and gibberellic acid (GA3) in the seeds, and the sensitivity of the seeds to these hormones were measured. The results of these experiments will be presented, and will aid our understanding of the interaction between genetics and maternal environment and their effects on seed dormancy. In turn, this will help us to better manage treatments for invasive weed species and to regenerate degraded land more effectively, by optimising the time of application of karrikinolide.
P0067 – ePoster
Gradients in grassland diversity on the pacific island
Alejandro Selkirk (Robinson Crusoe Archipelago,
Chile)
Greimler, J
P 4
1 , Gatica, A 2
, Penailillo, P 2 , Ruiz, E
, Baeza, C 3
3 , Stuessy, T
, Lopez, P
1
1 , Novoa,
1 University of Vienna, Faculty Center Biodiversity,
2
Systematic and Evolutionary Botany, Austria;
Departamento de Biologia Vegetal y Biotecnologia,
Universidad de Talca, Talca, Chile; 3 Departamento de
Botanica, Universidad de Concepcion, Concepcion,
Chile; 4 Jardin Botanico, Corporacion National Forestal,
Vina Del Mar, Chile
Grasslands on oceanic islands are among the ecosystems that are most severely altered by introduced biota. Both continental animals via herbivory, seed dispersal, and plants via competition have had a high impact on native plant communities. We focus here on gradients that can be observed in data collected in February 2011 on the island Alejandro Selkirk, situated c. 850 km west of
Valparaiso, Chile, in the southeastern Pacific. The grassland there covers a high proportion of the island especially in its lower elevations but ascending also to the summit area around 1,300 m. A decrease in number of weeds can be observed in horizontal and vertical distance from the only settlement and port. An ecological component is also present in the vertical gradient. This is expressed in a diversity depression in elevations around
1,000 where only few introduced species are found.
However, due to pseudo-alpine conditions around and above 1,200 m, the subantarctic (Magellanian) flora increases the grassland diversity again. The diversity depression and the fact that only a few introduced species comprise the grassland patches around 1,000 m may point to succession processes following disturbance by human impact (fire!) and the high (c. 3,000) goat population.
P0068 – ePoster
Additive partitioning in an oak forest diversity across
1 multiple spatial scales
Feng, Y 1
ICBR, China
Species distributions in a landscape are the result of processes operating at both local and regional spatial scales, measuring species diversity is critical for ecological research and biodiversity conservation.
Additive partitioning of three measures of diversity
(species richness, Shannon diversity index and Simpson diversity) was used to study the relationship between local and regional diversity across four spatial scales
(quadrat, slope-position, transect, and region) in an oak forest ( Quercus liaotungensis ), Beijing, China. Our observations suggest that local factors determine species abundance at the within-quadrat scale whereas patterns of richness are determined at large scales. Small scales are dominated by common species. Rare species represent a critical component of community and contribute to between-transect differences in community composition. The results of additive partitioning of the species-area relationship revealed that beta diversity due to factors such as among-patch heterogeneity, the surrounding matrix habitat, forest fragments, or chance effects have very different assemblages of species due to factors other than habitat area.
P0069 – ePoster
Genetic diversity of
Lilium tsingtauense
in China and
Korea assessed with ISSR markers and morphological characters
Guo, W 1,2 , Jeong, J 2 , Kim, Z 2 , Wang, R 1 , Kim, E 2 , Kim,
1
S 2
College of Life Science, Shandong University, Jinan PR
China; 2 College of Life Sciences and Biotechnology,
Korea University, Seoul, Republic of Korea
Genetic diversity and population structures of 12 populations with 648 samples of Lilium tsingtauense
Gilg (Qingdao Lily) in China and Korea were investigated using Inter-Simple Sequence Repeat (ISSR) molecular markers and morphological characters. At the species level, genetic diversity was relatively high, with the percentage of polymorphic loci 72.3%, effective number of alleles 1.437, average expected heterozygosities 0.231 and Shannon's information index
0.369, respectively. Considerable genetic differentiation
381

among natural populations (GST=0.144) and gene flow
(Nm=1.487) were detected. AMOVA analysis and
UPGMA-dendrogram suggested hierarchical regional structuring among the populations, and micro-scaled spatial structure appeared by spatial autocorrelation analysis. The morphological characters showed high correlation with ISSR parameters; No altitudinal differences were found and there was only a low genetic differentiation among different morphological types of L. tsingtauence in China. We therefore recommend in situ and ex situ conservation strategy of L. tsingtauense in terms of these results.
P0072 – ePoster
Phenotypic integration and endemism level of fourteen plant species in the Sub-Antarctic Kerguelen
Islands
Hermant, M
Hennion, F 4
1 , Prinzing, A 1 , Bailey, RI 2 , Bartish, I 3 ,
1 UMR CNRS ECOBIO, Rennes, France;
3
2 Evolutionary
Biology Centre, Uppsala, Sweden; Academy of Sciences of Czech Republic, Pruhonice, Czech Republic; 4 UMR
CNRS ECOBIO, Rennes, France
The sub-Antarctic region contains both many endemic species and strong abiotic gradients. It thus provides us with key opportunities to improve understanding of ecological and biological factors that may be involved in species geographical restrictions. Ecological explanations suggest that endemic species are specialized either on narrow or on particularly stressful habitats, both possibly linked to narrow phenotypic variability. However, variability per se may not suffice to reflect the interactions within and between complex phenotypes and complex environments. Here, we introduce phenotypic integration (i.e. number and strength of correlations between phenotypic traits) as a new explanation of endemism. We first examined the variation in mean traits
(growth, reproduction and reproductive strategies), and in phenotypic integration, between low and high altitude populations of an endemic species to the South Indian
Ocean Province, the Kerguelen Cabbage ( Pringlea antiscorbutica , Brassicaceae), at both small and large spatial scales. We then developed a multi-species approach with fourteen plant species present in the
Kerguelen Islands and with geographical distributions that ranged from highly endemic to cosmopolitan. We measured distribution, flowering phenology and multiple aspects of plant size along three types of abiotic environmental gradients in these species, and we compared their phenotypic integration, environmental integration and correlation between traits and environments. We found that changes in phenotypic integration with altitude in Pringlea antiscorbutica were as strong as changes in trait means, but did not occur at the same spatial scale: phenotypic integration showed higher variation at small spatial scales and much lower variation at large spatial scales. The increase in phenotypic integration with altitude at small spatial scales may restrict the responses to increasing stress with altitude at large scales to parallel shifts of all intercorrelated traits. Moreover, through the multispecies approach, we found that phenotypic integration, environmental integration and correlation between traits
382 and environments significantly increased with endemism level of species, suggesting that species with higher endemism levels may be constrained by particular combinations of environmental conditions, where these species are limited to particular combinations of phenotypic traits, resulting in strong trait/environment correlations. Overall, these results suggest that the more restricted the species area is, the higher its integration within and between phenotypes and environments is, likely resulting in a lower capacity of these species to respond to novel environmental conditions. Our results are particularly significant for the future of endemic sub-
Antarctic plant species under the current climate change in this region.
P0073 – ePoster
Responses of stomatal size and density to increasing
CO
2
vary within the southern Australian Myrtaceae
Hill, KE 1 , Watling, JR 1 , Hill, RS 1
1 University of Adelaide, Australia
Increasing
([CO
2 atmospheric concentrations of CO
2
]atm) are hypothesised to have opposite effects on stomatal density and size, decreasing the former and increasing the latter. These effects have been shown for many species across the Phanerozoic. Here we investigated the effects of recent rapid increases in
[CO
2
]atm on stomatal density and size of four
Myrtaceous species: Eucalyptus cneorifolia , Callistemon rugulosus , Leptospermum continentale and Melaleuca lanceolata . We used herbarium specimens and living material collected from small geographic ranges in South
Australia, thus reducing the effects of altitude and variability in climate, and enabling us to sample across a
130-200 year time period which corresponded with a change in [CO
2
]atm of 180–380 ppm. Different responses were observed across the four species.
Stomatal density decreased significantly for L.
continentale and M. lanceolata . Stomatal size increased significantly for E. cneorifolia and M.
lanceolata . No significant changes occurred for C. rugulosus stomata.
Stomatal conductance to water (gwmax), a value calculated from stomatal density and size, decreased significantly for all species except C. rugulosus , that showed marginal significance (P = 0.0808). The combination of fewer, larger stomata, such as seen with
M. lanceolata , results in lower conductance to both CO
2 and water. Due to the rise in [CO
2
]atm, intracellular CO
2 concentration increases, while transpiration decreases due to the reduction of stomatal conductance to water, the overall effect is to increase water use efficiency
(WUE: CO
2
fixed per H
2
O transpired). The consequences of these different responses to changing [CO
WUE of plants are discussed.
2
]atm for
P0074 – ePoster
Evolution of a hybrid zone of
Salix helvetica
and
S. purpurea
(Salicaceae) on the forefield of the Rhone
Glacier in the European Alps
Cosendai, A 1 , Dullinger, S 2 , Hadacek, F 3
Hoerandl, E 1
, Sagmeister, P 2 ,

1 University of Vienna, Dept of Evolutionary and
Systematic Botany, Austria; of Conservation Biology, Vegetation- and Landscape
Ecology, Austria;
2 University of Vienna, Dept
3 University of Vienna, Dept of
Chemical Ecology and Ecosystem Research, Vienna,
Austria
In the context of the global warming, new areas become available where glaciers have retreated and have left empty ecological niches for colonization. Such pioneer habitats are ideal for observation of evolutionary processes, especially if the time of retreat of the glacier is documented and recorded. During colonization previously isolated species are brought into contact, potentially resulting in hybridization. If hybrids can occupy extreme or intermediate habitats compared to the parental species, they eventually may establish populations as a first step towards speciation. To test this hypothesis, we analyze hybridization of two ecologically and morphologically distinct willow species, Salix helvetica and Salix purpurea , in the alluvial zone directly under the Rhone glacier in the Swiss Alps. We study population genetic diversity, genetic composition of the hybrid and putative backcrossing to the parents by use of microsatellite markers. The ecological niche of the two parental species and potential hybrid is assessed by vegetation relevees and analyses of soil types, preliminary results confirm frequent hybridization on the glacier forefield and differences in the parental ecological optimum. The assessment of soil characteristic and types of vegetation suggest that the hybrids have a broad ecological amplitude, but occupy preferentially habitats that combine characteristics of the parental species. Further studies on population genetic structure, backcrossing and fertility will give insights whether the hybrids do have a potential for speciation.
P0075 – ePoster
Conservation of critically endangered plant species based on information obtained by the ubiquitous
1 genotyping
Isagi, Y 1 , Kaneko, S 1 , Kato, K 1
Shiga, T 2 , Masumoto, I 3 , Ohtake, K
Kyoto University, Japan;
History, Japan; 3
2
, Izuno, A
3
1 , Mizutani, M 1 ,
Osaka Museum of Natural
Chuden Engineering Consultants,
Hiroshima, Japan
As is common in most developed countries, many plant species have become threatened with extinction, for example, in Japan about 24% of vascular plant species are classified as endangered. Because every individual of organisms have a finite lifespan, regeneration processes of populations must be considered in order to conserve threatened species in the long term. Genetic information can provide crucial information about regeneration processes such as clonal structure, gene flow, inbreeding depression and relatedness. In addition, genetic information provides data on genetic diversity and structure which are important for conserving species.
However, many problematic activities have been undertaken for the purpose of biological conservation due to the lack of or complete absence of genetic information. These problems, for example, include propagation and reintroductions that do not represent the full genetic diversity of species, transplantation without consideration of intraspecific genetic differentiation, breeding without consideration of relatedness, hybridizing with related species that causes genetic pollution, etc. These activities, though most of them have been done with good intentions, may in fact cause fatal effects on the endangered plant species. We selected 50 critically endangered plant species whose remnant populations number less than 1,000 individuals, then, determined the habitats and genotypes at microsatellite loci for all plant individuals growing in the wild. We call this research approach Ubiquitous Genotyping. By taking advantage of information from ubiquitous genotyping, we can evaluate genetic traits of plants, select appropriate plant individuals for artificial crossing, determine appropriate locations for transplantation based on genetic structure, detect genetic pollution, prevent illegal trading by genetic tagging, and evaluate the long term viability of populations. In this paper, we will provide tangible examples of how biological conservation can be aided by incorporating information from ubiquitous genotyping for several critically endangered plant species:
Adenophora palustris ( Campanula ceae, Japan, Korea and China, n = ca. 800 in the wild of Japan),
Veronicastrum villosulum (Plantaginaceae, Japan and
China, n = ca. 130 in the wild of Japan), Meterosideros boniensis (Myrtaceae, endemic to Japan, n = ca. 330 in the wild), Stachyurus macrocarpus var. prunifolius
(Stachyuraceae, endemic to Japan, n < 20 in the wild),
Loropetalum chinense (Hamamelidaceae, Japan, China and India, n = ca. 100 in the wild of Japan), Nuphar submerse (Nymphaeceae, endemic to Japan, n = ca. 100 in the wild) and Athyrium viridescentipes (Woodsiaceae, endemic to Japan, n = ca. 100 in the wild). This research was supported by the Environment Research and
Technology Development Fund (D-0903) of the Ministry of the Environment, Japan.
P0076 – ePoster
Conservation of a critically endangered tetraploid
1 fern species
Athyrium viridescentipes
based on the ubiquitous genotyping approach
Izuno, A 1 , Takamiya, M 2 , Kaneko, S 1 , Isagi, Y 1
Kyoto University Graduate School of Agriculture,
2
Division of Forest and Biomaterials Science, Japan;
Kumamoto University Graduate Schhol of Science and
Technology, Division of Life Science, Japan
Of the c. 750 fern species described in Japan, 185 are threatened with extinction. Understanding the genetic and ecological characteristics of these fern species is important for developing effective conservation management programs. However, in general, little information is available concerning the genetic diversity and structure of fern species. In particular, few study focused on conservation genetics of ferns. Athyrium viridescentipes Sa. Kurata (Woodsiaceae) is a critically endangered hypotetraploid (2n = 156) fern endemic to southern Japan. The remnant populations are highly fragmented and the number of individuals is extremely small (n = 103). Geographic distances among the four remaining populations are between 2 km to 120 km, with the number of individuals in each population being 5, 5,
34, and 59, respectively. In order to elucidate the genetic
383

status of A. viridescentipes , we conducted ubiquitous genotyping, by determining the genotypes of all 103 known individuals at 15 nuclear microsatellite loci. In addition, the genotype of 20 spore from one individual was determined to reveal the breeding system (sexual vs. asexual) of this species. The level of allelic variation of
A. viridescentipes was remarkably low, with the average number of alleles per locus (A) being 1.8. This value was smaller than those of three closely related tetraploid
Athyrium species. Although the four populations are geographically isolated, the genetic composition among populations were quite similar with the FST values among four populations being approximately 0.01. Sixtynine individuals (67%) showed an identical pattern of alleles at the 15 microsatellite loci suggesting that clonality may occur in the species. However, the allelic combinations of all 20 spore genotyped were different from those of their parent sporophyte indicating that the species can undergoe sexual reproduction. While the genetic variation is very limited at present different genotypes may occur in future generations through recombination. In order to maintain and conserve the present four populations, we have to monitor the population size and genetic variation continuously. This research was supported by the Environment Research and Technology Development Fund (D-0903) of the
Ministry of the Environment, Japan.
P0077 – ePoster
Clonality, polyploidy and sterility in two rare species of
Grevillea
(Proteaceae)
James, E 1,3 , McDougall, K 2 , Ladiges, P 3 , Newbigin, E 3
1 Royal Botanic Gardens Melbourne, Australia; 2 Dept of
Environment, Climate Change and Water, Australia;
3 School of Botany, University of Melbourne, Australia
Clonality is a reproductive strategy common to many plants. It provides a mechanism for persistence in the landscape when environmental conditions are hostile to sexual reproduction although recruitment from sexually derived seed is considered necessary to maintain genetic diversity and long-term viability. For the majority of clonal plants, reproduction is a combination of both vegetative and sexual reproduction with the relative contribution varying with environmental conditions. In the case of two rare species of Grevillea from southeastern Australia, clonal growth appears to be the sole method of reproduction. One species, G. infecunda , is male sterile. The second species, G. renwickiana , produces a large number of floral buds but rarely flowers and seed production has not been seen. Spatial genetic analysis using microsatellites confirmed clonality in both species. Clones tended not to intermingle although fragmentation of extensive, presumably long-lived clones has occurred. Fewer than 30 genotypes were identified for each species. Based on chloroplast sequencing, each species comprises fewer than ten maternal lineages. The role of polyploidy and somatic mutation in the sterility, genetic diversity, distribution and persistence of the species will be discussed in relation to conservation.
384
P0078 – ePoster
Online mapping of botanical resources for Hawaii and New Guinea: test cases for the Pacific Basin
James, S 1 , Masaki, D 2 , Allison, A 1 , Pyle, R 1
1 Pacific Biological Survey, Bishop Museum, Honolulu,
Hawaii; 2 USGS National Biological Information
Infrastructure, Pacific Basin Information Node, USA
Museum specimen data are essential for taxonomic and biodiversity research, invaluable for mapping historic and predicted distributions of native and introduced species for conservation and ecosystem restoration activities, and as an educational tool. A collaborative effort between the USGS National Biological
Information Infrastructure’s (NBII) Pacific Basin
Information Node (PBIN) and the Bishop Museum is producing a suite of web-based mapping tools using
Adobe Flex and ESRI ArcGIS Server. The effort leverages the extensive digital collections and taxonomic expertise of Bishop Museum staff along with the geospatial technical capacity of the USGS Pacific Basin
Information Node. The resulting map service brings together species occurrence elements, high resolution aerial imagery and ecological attributes (climate, soil types, etc) to provide users with a visual data interface for query and navigation. In addition to the spatial viewer, a web portal providing species information featuring Pacific Region specimen, taxonomic data and digital images is available online at http://nsdb.bishopmuseum.org and is cross-linked to the
Integrated Taxonomic Information System (ITIS - www.itis.gov). Current efforts are using data from the
Hawaiian Islands and New Guinea collections for an initial pilot project which will be expanded to provide mapping and taxonomic services for the Pacific Basin.
P0080 – ePoster
Conservation of anticancer drugs yielding medicinal plants in the east coast of Tamilnadu, India
1
John De Britto, A 1
St Xavier's College (Autonomous), Palayamkottai,
Tirunelveli, India
Cancer is the killer disease. Discovery of much effective and less toxic drugs and treatment are the national and international priorities of medical research. In general, medicinal plants in the coastal areas are characterized by low population densities, greater inter population and intra specific variations. The medicinal plants found in coastal areas possess stress-induced metabolites belonging to chemical groups such as steroids, triterpenoids, saponins, flavanoids, alkaloids and tannins.
These chemicals are mostly unexplored for anticancer property. Accuracy in recording the medicinal use of a plant, molecular and chemical characterization, their roles and selection of the superior genotypes of the plant species are important issues that need to be verified in the development of drugs of plant origin for cancer. Such medicinally valuable plants in the coastal areas should be conserved. Two anticancer drug yielding plants of the
East coast of Tamilnadu Abrus precatorius L., and

Pongamia pinnata Pierre were collected from five locations in the east coast of Tamilnadu, India. Natural and anthropogenic factors were investigated. Molecular variations using ISSR markers were analyzed.
Identification of genetic polymorphism was done using
Pop Gene 3.2 and NTSYS. Phytochemical (active principles) variations in these medicinal plants using
HPLC were analyzed. Considering the genetic polymorphism and the quantity of active principles, superior genotypes were identified. Strategies for conservation of these plants are discussed. Natural factors such as unprecedented climatic changes and anthropogenic pressures such as habitat degradation and over harvesting are largely responsible for genetic depletion and rarity of these medicinal plant species.
New means and mechanisms are to be worked out for long-term conservation and sustainable utilization of these rare and economically important medicinal plants.
P0081 – ePoster
Ex situ conservation: the challenge of Brazilian
Botanical Gardens in achieving target 8 of the global strategy for plant conservation
Judice, D 1 , Moraes, M
Martinelli, G 2
1 , Messina, T 1 , Martins, E 1 ,
1 National Centre For Plant Conservation, Brazil; 2 Rio de
Janeiro Botanical Garden, Brazil
Ex situ conservation is a much needed tool considering the present state of biodiversity depletion and the urge to guarantee the persistence of threatened plant species outside natural populations. Target 8 of the Global
Strategy for Plant Conservation (GSPC), calls for 60% of threatened plant species cultivated in accessible ex situ collections and 10% of them included in recovery and restoration programs. The GSPC target was indeed a starting point to the implementation of ex situ conservation programs and to the elaboration of protocols and guidelines to be adopted by the parties. On the other hand, developing countries, including Brazil, still face restrictions and limitations in accomplishing ex situ conservation projects, such as fund raising and institutional infrastructure, as well as data insufficiency of flora at a national level. Brazil is a mega diverse country with approximately 20% of the global flora
(40.989 species), which makes even harder to consolidate an efficient ex situ conservation strategy and to achieve the 60% proposed by GSPC. Farther on, in situ conservation has been carried out but has not been truly effective over the past decades, since only c.10% of the
Atlantic Forest is remaining. Therefore, ex situ conservation must also be implemented and Botanical
Gardens may play an important role. This survey aims to analyze all quantitative data of ex situ living collections of threatened species existent in Botanical Gardens of
Brazil, as well as seed banks across the country, in order to attain an updated figure of the reality. There are 30
Botanical Gardens in Brazil, where only five of them are registered as Botanical Garden Conservation
International (BGCI) members. This numbers justify the quantitative results showed by a recent research carried out by the Jardim Botânico do Rio de Janeiro (JBRJ) through the Rede Nacional de Jardins Botânicos do
Brasil (RNJB), in which 81 taxa of 22 botanical families can be found in 11 Botanical Gardens as ex situ living collections, representing 17% of the 472 species listed on the official list of Brazilian threatened flora. Scientific researching and conservation of threatened plant species throughout ex situ programmes and collections are mandatory in order to maintain diversity and enlarge species surviving rates.
P0082 – ePoster
The Brazilian list of flora
Martins, E 1 , Moraes, M 1 , Messina, T 1 , Judice, D 1 ,
1
Martinelli, G 2
Brazilian National Centre for Plant Conservation,
Brazil; 2 Rio de Janeiro Botanical Garden, Brazil
The Global Strategy for Plant Conservation – GSPC – establishes as its first target that a ‘widely accessible working list of known plant species’ must be consolidated by the parties with the aim of understanding and documenting plant diversity, ‘as a step towards a complete world flora’. Plant inventories in mega diverse countries are a challenging task due to its great species richness, the lack of information in scientific collections and the poor networking among scientists and institutions. In order to mainstream national efforts in
Brazil with international initiatives towards plant conservation, the Brazilian National Centre for Plant
Conservation – CNCFlora, was created under Rio de
Janeiro Botanic Garden Research Institute – JBRJ infrastructure. The Centre`s mission is to coordinate national efforts, and to understand, document, and conserve plant diversity, in collaboration with research institutions and environmental agencies. The actions are being organized accordingly to GSPC framework, prioritizing targets that meet national needs and capacities. Target one has always been considered a challenging task, given the diversity of the Brazilian flora. The last compilation of plant species was published in the 19th century, edited by Von Martius, Eichler and
Urban and named Flora Brasiliensis . After that, several initiatives have failed. Since 2008, JBRJ has been working on the development of the Lista de species –
Flora do Brasil. For that, CNCFlora has articulated 480 researchers from national and international institutions through an online platform, specially designed in collaboration with the Centro de Referência em
Informação Ambiental – CRIA. The process used the
Species2000 protocol, and generated a species list composed by 40,989 taxa (3.608 fungi, 3.496 algae,
1.521 bryophytes, 1.176 pterodophytes, 26 gymnosperms e 31.162 angiosperms). The biome with the highest number of species was Mata Atlântica (19.355 species), followed by Amazônia (13.317 espécies), Cerrado
(12.699 espécies), Caatinga (5.218 espécies), Pampa
(1.964 espécies) and Pantanal (1.240 espécies). The list can be assessed online, and the website also contains information such as the scientific and common name of a species, synonyms and distribution. This represents a major achievement for the Botanical Community, since a huge proportion of the worlds plant diversity occurs in
Brazil and is under constant threats due to unsustainable land use practices. The data base will be updated systematically, in order to keep track of taxonomic changes. The catalog is also expected to be used for
385

supporting development of conservation policies and for standardizing taxonomic decisions. Besides compiling all flora species, the List will also be used as a taxonomic guide for the elaboration of the Official List of
Threatened Plant Species, GSPC’s target two.
P0083 – ePoster
Field data and conservation biology in Brazil: the need of a more integrative approach in order to conserve flora endangered species
Messina, T 1 , Moraes, M 1 , Judice, D 1 , Martins, E 1
1
Martinelli, G 2
National Centre For Plant Conservation, Brazil; 2
,
Rio de
Janeiro Botanical Garden, Brazil
Mega diverse developing countries face a difficult task when dealing with conservation of flora threatened species. Limited financial means for conservation projects such as researching, sampling and monitoring of protected areas, create great obstacles that make a proactive approach on the subject, a remote reality. Also, developing countries have the imminent need of improving their scientific information systems and enhancing quality of biological collections. The poor data base of species information existent, the lack of scientific networking and the traditional culture existent among scientists, conditioned to only get the minimum information or the only information needed to reach their own purposes while doing their field work, are a step back for flora conservation in a country with such large proportions and diversity. In order to guarantee effective species conservation, considering that Biodiversityrelated sciences, due to their complexity and youth, have a major challenge to face and not much time available, efforts from all different knowledge fields must acquire a more integrative approach enabling the success in the bigger picture, which is saving species from extinction.
By doing that, all field Biologists can contribute to threatened species conservation studies and actions, even if indirectly, therefore a collaboration in order to acquire a single vision towards the improvement of strategies for such an intrinsic matter as halting extinction risk have to be put in practice. The aim of this work is to propose the minimum necessary information to be collected on a field trip by a plant scientist of any expertise area and to point out the need of an integrative approach in order to achieve conservation goals and standardize on a national level, the gathering of field information. It is mandatory to understand that the main goal is to upgrade the quality of information avoiding by that rework and the loss of precise data that can be used for different matters and subjects. The information proposed can be incorporated to Herbarium cards suggesting the different data to be collected such as: identification of incident threats, habitat, occurrence of alien species in the study area, as well as the occurrence of other subpopulations in the area and number of individuals. Farther more, the GPS has been available in the market since the sixties, but there are a large number of plant samples without an accurate coordinate. Recording precision will not only improve the quality of work as it can be used for a variety of studies, as Prediction Modeling and Ecology, among many others. Building and compiling more detailed data on diversity and pointing out the necessity of
386 multidisciplinary approaches enabling conservation of flora threatened species will be a major step for science and for flora threatened species conservation.
P0084 – ePoster
Cryopreservation of native Western Australian flora to aid post-mining restoration
Kaczmarczyk, A 1, 2
Smith, S 4
, Funnekotter, B 0
, Mancera, RL 1
, Bunn, E 0 , Che, P 4 ,
1 Curtin University, Bentley, Australia; 2 Botanic Gardens and Parks Authority, West Perth, Australia; 3 School of
Plant Biology, University of Western Australia, Crawley,
Australia; 4 Australian Research Council Centre of
Excellence in Plant Energy Biology and Centre of
Excellence for Plant Metabolomics, University of
Western Australia, , Australia
Mining in Western Australia is predicted to rapidly expand, an outcome that will require effective land restoration to reassemble functional native ecosystems with high species abundance and diversity following mining. Alcoa Australia and BHP Billiton Worsley
Alumina currently use several different methods to return the majority of native species to their mine sites.
However, for a small group no effective method of conventional propagation is currently viable limiting propagation options to plant tissue culture. Loxocarya cinerea (Restionaceae) is a locally abundant understorey
‘native rush’ species in the Western Australian jarrah
( Eucalyptus marginata ) forest where mining for alumina is prevalent. This species is important in post mining restoration as it is resistant to dieback disease
( Phytophthora cinnamomi ) and forms large rhizomatous clonal colonies with soil binding properties that reduce soil erosion. Loxocarya cinerea is dioecious, rarely produces seeds and conventional propagation extremely difficult, therefore it is by necessity micropropagated.
However continuous maintenance of multiple provenance culture lines incurs risks of unwanted somaclonal variation and losses due to accidental culture contamination – leading to increased costs, hence the current study on cryopreservation of L. cinerea for longterm cost-efficient storage of valuable culture lines.
Shoot tips are used commonly for cryopreservation, which is the storage of viable material at temperatures of liquid nitrogen (-196°C). Thus far cryogenic experiments have met with limited success with L.
cinerea highlighting the current problems with the development of new cryogenic approaches for poorly understood species. Currently there is an inadequate understanding as to why one cryogenic protocol works for one species or even genotype but not for another. The initial aim of this study is to develop a suitable cryopreservation protocol for L. cinerea and other native species of interest to the mining industry. The second aim is to develop a detailed understanding of the underlying species physiology through biochemical analyses on shoot tips to compare membrane and sugar compositions of cryotolerant and cryosensitive species. Preliminary results have found clear differences in phospholipid, sterol and sugar compositions between L. cinerea ,
Grevillea scapigera (a critically endangered species) and
Arabidopsis thaliana – a model ‘cryo-friendly’ species used in this project for comparison purposes. Notable

differences within species were also found after various cryogenic pretreatments that were also shown to influence post cryogenic survival. Survival following cryopreservation was highest in A. thaliana , shoot tips
(87.5%) compared to G. scapigera (51.5%) or L. cinerea
(<5 %). However, the total sterol and phospholipid content increased for all three species following alternating temperature (AT) ‘pre-conditioning’ treatments. Whereas the total soluble sugar concentration decreased in A. thaliana after AT, G. scapigera and L. cinerea showed increases in total soluble sugars after
AT. Further experiments aimed at analysing the state of water within shoot tips using differential scanning calorimetry (DSC) have also helped to establish correlations between cryopreservation results and biochemical compositions, which is increasing our overall understanding of the cryopreservation process and facilitating the development of novel approaches for poorly studied species requiring cryopreservation.
P0085 – ePoster
Conservation of a critically endangered shrub species
Stachyurus macrocarpus var. prunifolius
(Stachyuraceae) based on the ubiquitous genotyping approach
Kaneko, S 1 , Abe, T 2 , Isagi, Y 1
1 Kyoto University, Japan;
Research Institute, Japan
2 Forestry & Forest Products
Approximately 3,000 species are listed as critically endangered in the world. Conservation measures such as restoration of habitat, assisted reproduction, and reintroduction is required to prevent the extinction of critically endangered species in the wild. However, genetic and ecological knowledge about endangered species are usually very limited, and this limitation might lead to the conservation measures being ineffective.
Here, we propose that ubiquitous genotyping, that is, a method whereby the genotype for all known wild individuals of the species is determined using high resolution genetic markers, is an essential method required to the effective conservation management for critically endangered species. In this study, we undertook ubiquitous genotyping of Stachyurus macrocarpus var. prunifolius endemic to the Ogasawara (Bonin) Islands in
Japan. We determined the genotype of all 15 individuals known in the three wild populations using microsatellite markers. In addition, the seeds of one wild individual were collected to form the build of an ex situ collection, and 60 seedlings were genotyped in total. The results of ubiquitous genotyping showed that both wild and nursery populations contained the population specific alleles.
Two alleles only detected from the nursery population indicate the existence of as yet undiscovered mature individuals in the wild population. Four alleles specific to the wild population were detected in two individuals isolated from the main wild populations. These isolated individuals are therefore of high conservation priority to maintain intra-specific genetic diversity of the species. In addition significant heterozygote excess was observed in the wild population individuals. This heterozygote excess may suggest the existence of inbreeding depression, and indicates the need to evaluate the factor causing inbreeding depression in the wild environment prior to reintroductions. These results show that ubiquitous genotyping data can provide essential genetic and ecological information that is essential for effective management of critically endangered species. The present findings also warn of the inefficiency of conservation measures without the knowledge of genetic characteristics of all known individuals. Ubiquitous genotyping should be considered as an integral component of conservation programs for critically endangered species. This research was supported by the
Environment Research and Technology Development
Fund (D-0903) of the Ministry of the Environment,
Japan.
P0089 – ePoster
Conservation of a critically endangered perennial species,
Veronicastrum villosulum
(Plantaginaceae) by ubiquitous genotyping
Kato, K 1 , Kaneko, S 1 , Isagi, Y 1
1 Kyoto University, Kyoto, Japan
For effective conservation management of threatened plants, we must design conservation programs with consideration of genetic information. For example, it is important to ensure that ex situ collections contain an adequate representation of the species genetic diversity.
Veronicastrum villosulum (Miq.) T. Yamazaki is a critically endangered perennial herb rediscovered in southern Japan in 1989, with only one known population
(area equals ca. 100m2). Plants derived from this wild population have been cultivated in local elementary schools, while plants of unknown origin have been cultivated in a few botanical gardens in Japan. The objective of this study was to reveal the genetic diversity and structure of V. villosulum in the wild, to understand the genetic composition of cultivated individuals and to apply this knowledge to the future conservation management of the species. A total 256 individuals of V. villosulum were collected and the genotypes were determined using nine nuclear microsatellite markers developed for this study. The numbers of samples were as follows: 139 individuals from the single wild population, 99 individuals from the elementary school collection and 18 individuals from three botanical gardens in Japan. Microsatellite analysis of all 256 individuals revealed 13 multilocus genotypes. We found that the wild population of V. villosulum was composed entirely of one clone. Individuals from the elementary school collection contained (1) the same wild clone
(83%), (2) nine clones originating from self-fertilization of the wild clone (10%) and (3) two unique clones (7%).
The cultivated individuals in three botanical gardens all consisted of one unique clone. The finding of three clones not in the wild population implies that some wild plants may have become extinct since collection. In order to conserve this genetic diversity of wild and cultivated plants an ex situ collections containing all four unique clones was established at the Hiroshima botanical garden in November 2010. The clonal identity of individuals of this species can be determined by genetic tagging using the nine microsatellite loci developed for the species in the future. These approaches will contribute to the conservation of V. villosulum. This research was supported by the Environment Research and Technology
387

Development Fund (D-0903) of the Ministry of the
Environment, Japan.
P0090 – ePoster
Using structural complexity as a surrogate measure for biodiversity
Keeton, K 1,3 , Davidson, N 2 , Bailey, T 3 , Close, D 4
1
3
University of Tasmania, Australia; 2 Greening Australia;
CRC Forestry, Hobart, Tasmania, Australia; 4 TIAR,
Tasmania, Australia
The ecosystem services provided by native forest are becoming valued in Australia and a price on carbon is soon to be decided. The international price placed on carbon (Chicago open market) is $10/t. Biodiversity may be the next ecosystem service to be traded. Already traders are talking about the sale of biodiverse carbon
(carbon grown using a biodiverse mixture of local native species). However there is no practical and economical measure of biodiversity currently available. Therefore this project sets out to compare two measures of structural complexity as surrogates for biodiversity. We assessed 40 sites in the midlands of Tasmania, Australia
– 20 sites were healthy native forest remnants and 20 sites were block plantings of local native species
(restoration plantings) established between 6 and 25 years previously. Using the McElhinny quantitative index of structural complexity (McElhinny et al 2006), restoration plantings were shown to have a low percentage score (50%) compared to healthy native forests (70%), but a higher score than farmland (40%).
Habitat Hectares (Parkes et al 2002), which is a tool for assessing stand condition, provides a qualitative measure of structural complexity and landscape-scale connectivity. When the same sites were assessed, this measure gave a higher relative score to restoration plantings. Direct measure of biodiversity using bird surveys, conducted for half an hour starting at dawn in summer on a subset of 10 sites (5 restoration sites and 5 healthy native forest sites) were in agreement with structural complexity measures. This project demonstrates that structural complexity does provide a means by which the ecosystem service biodiversity can be valued and sold.
P0091 – ePoster
Developing a state of the environment indicator for long term monitoring of the impacts of climate change on Antarctic terrestrial communities
1
King, D 1
Lucieer, A
, Wasley, J 2 , Ryan-Colton, E 1
3 , Robinson, S 1
Antarctic Division, Kingston, Australia;
Tasmania, Hobart, Australia
, Turnbull, J
University of Wollongong, Australia;
3
2
1 ,
Australian
University of
Climate change is occurring rapidly at high latitudes, yet for East Antarctica few studies have monitored the impact of environmental change on terrestrial vegetation.
Severe impacts are predicted if climate change affects water availability, a key driver of Antarctic terrestrial dynamics. Long-term monitoring of vegetation
388 communities along a moisture gradient at two sites in the
Windmill Islands, East Antarctica has commenced, forming Antarctic State of the Environment Indicator 72.
Three complementary sampling regimes were implemented in both 2003 and 2008; turf water content, broad scale percent cover of vegetation types and finer scale relative abundance of bryophyte species and lichen taxa. The baseline data reveal a transition from bryophyte to lichen communities with increasing aridity. For bryophyte species, the relatively desiccation sensitive
Schistidium antarctici dominates the wettest communities, whilst the desiccation tolerant Ceratodon purpureus is most abundant in the drier transitional communities. Previously known physiological tolerance of species and vegetation types to water availability allows prediction of vegetation shifts under different climate scenarios, for example expansion of C. purpureus into formerly wetter communities if the climate dries. Digital image analysis was trialled, as an alternative method to visual percent cover estimation, in an attempt to improve survey methodology in these extreme field conditions. Both methods estimated cover to within 5%, although digital methods underestimated scarce vegetation types in complex lichen communities.
In 2008, field confirmation of scarce vegetation types supplemented digital imagery. Currently, new methods of mapping moss beds, using Unmanned Aerial Vehicle
(UAV) aerial photography to provide ultra-high resolution spatial data, are enabling high resolution modelling of environmental factors affecting moss distribution and growth. Our results indicate that this methodology is sensitive enough to detect climateinduced change in these slow growing communities.
P0092 – ePoster
In vitro culture enables the propagation of ecological
1
J 1 keystone sedge species
Kodym, A 1 , Turner, S 2 , Bunn, E 2 , Temsch, E 3 , Delpratt,
University of Melbourne, Richmond, Australia; 2 Kings
3
Park and Botanic Garden, West Perth, Australia;
University of Vienna, Vienna, Austria
Many Australian character species cannot be returned to restoration sites because of the lack of efficient propagation methods. These include several
Lepidosperma and Gahnia species (Cyperaceae) that possess unresolved dormancy, often low seed fill and are not responsive to vegetative propagation by division. In vitro techniques are being investigated as a means of producing large quantities of planting material while retaining appropriate levels of genetic diversity. Seeds of
Lepidosperma concavum , L. laterale and L. longitudinale , Gahnia trifida and G. filum germinated in vitro after removal of the pericarp and successfully established in the nursery. Germination ranged from 60-
91% depending on the species and population and occurred within 7 to 18 days on 1/2MS media supplemented with 1 µM zeatin and 10 µM GA3. In vitro plants could be successfully stored without subculturing at 14˚C under low light conditions for at least one year.
Micropropagation in L. laterale was initiated using shoot culture. Micropropagation rates were 1.3–2.8 over a 6 weeks period on medium containing BAP while shoots

rooted easily on auxin medium. As a tool for mass propagation and in preparation for the production of artificial seed, somatic embryogenesis was initiated.
Embryogenic callus was induced from immature seed and from in vitro-grown seedlings on medium containing
2,4-D in combination with zeatin. Induction was 16% and 100% respectively within 6 weeks. High rates of conversion to plants were achieved on charcoal medium followed by TDZ medium. Acclimatisation of rooted plants was 95%. Acclimatised plants grew vigorously under standard nursery conditions. Flow cytometer measurements determining ploidy level were carried out on plantlets regenerated from seed derived callus.
P0093 – ePoster
1
Comparisons of endangered plants between Japan and adjacent East Asian regions
Kokubugata, G 1 , Kato, M 1
National Museum of Nature and Science, Japan
Previously many studies and investigations have been performed for endangered species within countries or regions of East Asia. These studies and investigations are very important and valuable for understanding endangered species at the national and regional level, but also we believe that global concept must be more applied for biologically considering these endangered species because national borders are no mean for wild animals and plants. Here, we numerically compare endangered vascular plants between Japan and four adjacent East
Asian regions, namely China, South Korea, eastern part of Russia and Taiwan; and try to suggest essential matters for globally considering endangered plants in
East Asia. In 2007, a Red List of Japanese endangered vascular plants was published by the Japanese Ministry of Environment. Following the list, 2,018 endangered plant taxa (including EX, EW, NT, DD) exist in Japan.
On the other hand, 571 and 1,335 plant taxa are respectively listed in a red list of South Korea (Korean
National Arboretum, 2009) and in a combined Red List based on nine areas of eastern part of Russia
(Khabarovsk, Primosrky, Amur, Sakhalin, Yakutia,
Kamchatka, Buryat, Irkutsk and Chita). In China and
Taiwan, the newest plant Red List is under preparation
(Wang and Xie, prep.; Taiwanese Council of Agriculture, prep.). Between Japan and the four regions of China,
South Korea, eastern part of Russia and Taiwan, 93, 93,
88 and 91 taxa are respectively counted as common endangered plants. Out of the common endangered plants, Orchidaceae is most found between Japan and three regions of China, South Korea and Taiwan; and
Cyperaceae is most found between Japan and eastern part of Russia. On the other hand, it is noteworthy that there are no plant taxa being commonly treated as endangered species in all of five regions. In comprehensively understanding endangered plants in East Asia, we should: 1) push forward with global taxonomic studies, because taxonomic treatments, being generally accepted at present, of these endangered plants may not always be correct, especially, there is a possibility to miss endemic plants in each region; 2) investigate distribution ranges and in situ situations of each plant taxon in East Asia, because it is not necessary that all of endangered plants in a certain region are on the edge of extinction in other regions; and 3) standard categories and criteria for sharing an uniform concept of endangered plants in East
Asia.
P0094 – ePoster
The capacity of
Echium plantagineum
(Paterson’s
Curse) to evolve and respond to predicted drought conditions
Konarzewski, T 1 , Godfree, R 2 , Murray, B 3
1 CSIRO and UTS, Sydney, Australia;
Plant Industry, Canberra, Australia;
2 CSIRO Division of
3 Plant Functional
Biology and Climate Change Cluster, Dept of
Environmental Sciences, University of Technology
Sydney, Australia
The exotic plant species Echium plantagineum is responsible for substantial ecological, economic and social problems across much of Australia. E. plantagineum causes substantial ecological damage through the invasion and degradation of native grassland, high cost of control in agricultural regions due to the degradation of land and poisoning of livestock which has severely influenced Australian meat and wool industries.
Each plant is capable of releasing up to 10,000 seeds which can remain in the soil for up to ten years requiring expensive large scale control programs. The key objective of our research project is to experimentally determine variation for morphological and demographic traits associated with drought tolerance over a range bioregions spanning a temperature and rainfall gradient, and to link these data with the evolutionary potential of
E. plantagineum under future climatic changes.
Predicting the response of an invasive species to climate change will; assist in determining the future distribution of this species, provide a model to predict the changes in distribution of other native or invasive broad leaf species, allow land managers to allocated their resources accordingly and assist industry to make appropriate decisions regarding control methods. The capacity to respond to climatic changes is species and trait dependant and it is important to know if invasive species have this capacity to evolve. Evolution comes at a high cost and plants are only able to respond as long as they have sufficient resources and as Australian soils are already resource limited species may not have the necessary resources to evolve. Seed samples were collected in 2009 from 50 plants at each of 34 sites within six bioregions along a 1000km temperature and rainfall gradient representative of E. plantagineum habitats in eastern
Australia. We investigated variation in seed weight, seed dimensional characteristics, seed germination rates and seed viability across and within each collection location.
A large scale glasshouse-based growth experiment was conducted during the 2010 growing season where established seedlings were measured for geographic variation in morphological, physiological and drought tolerance traits across and within populations. This work will provide information on the evolutionary ability of this species to adapt to climate change leading to insight into its future distribution thus allowing land managers to appropriately adjust their management strategies and resource allocations to control this invasive species.
Future work will focus on integrating glasshouse-based trait data with evolutionary potential and impacts on
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native plant communities under field conditions. Invasive species are a global problem that requires research and development to ensure that the most ecologically safe and cost effective methods are used. To be able to control these species we must know how they will react to future climatic changes and where we should focus our resources.
P0095 – ePoster
Hanging gardens at risk – land-use impact on epiphyte diversity in two Ecuadorian rain forest areas
Köster, N 1, 2 , Nieder, J 1 , Barthlott, W 1
1 Nees Institute for Biodiversity of Plants, University of
Bonn, Germany; 2 Botanical Garden, University of
Tübingen, Baden-Württemberg, Germany
Tropical rain forests are strongly threatened by human land-use. As outstanding elements of these forests, vascular epiphytes are likely to be affected substantially by forest loss and fragmentation. This study analyzes the effect of deforestation on epiphyte diversity and the potential for epiphyte conservation in anthropogenically transformed habitats. It is based on the comparison of epiphyte species numbers of 220 study plots in primary forests, forest fragments, isolated remnant trees in young and old pastures, as well as young and old secondary forests. Field work was conducted at a lowland rain forest and a cloud forest study site in western Ecuador, one of the global centres of biodiversity. Accordingly, epiphyte diversity of both study sites was remarkably high, comprising 381 and 383 species, respectively. The study revealed a pronounced decrease in epiphyte species numbers from primary forests through forest fragments to isolated remnant trees and secondary forests, although species numbers increased considerably with maturing of secondary forests. Species assemblages on isolated remnant trees impoverished distinctly with time since isolation and may serve for recolonization of secondary forests only for a short time. Spatial patterns of habitat fragmentation had no significant effect on epiphyte diversity, at least within the time frame and spatial range studied. Host tree characteristics explained a remarkable amount of the variation in epiphyte diversity. For isolated remnant trees, host tree genus statistically explained up to half of the variation in epiphyte species richness. Tree size proved to be a core predictor of species richness among secondary forests, but had less significance within other habitat types. Epiphyte diversity is largely driven by the fact whether a forest is mature, regrown after complete clear-cutting, or heavily reduced to forest fragments and isolated remnant trees. However, the structural and taxonomic composition of the tree community in these habitat types is also fundamental to epiphyte diversity. Differences in the response to habitat transformation between species indicate an important role of life-history traits and taxon-specific ecological constraints. Araceae and Hymenophyllaceae showed the highest decrease of species numbers in transformed habitats, whereas Bromeliaceae even increased slightly on isolated remnant trees. Secondary forests were dominated by facultative epiphytes and hemi-epiphytes, which are often confined to the tree trunk. Isolated remnant trees harboured mostly epiphytes typical for tree crowns, with high proportions of drought tolerant
390 species. Dispersal mode had little influence on the response to habitat transformation, indicating a dominance of microclimatic constraints compared to dispersal limitations. Epiphytes proved to be a suitable model group for studying human impact on forestdwelling life forms. Isolated remnant trees and secondary forests can harbour a remarkable part of regional epiphyte floras, but temporal dynamics in these habitats emerged to be a major driver of epiphyte species richness: epiphyte assemblages on isolated remnant trees gradually impoverish and their potential for epiphyte conservation is, thus, temporarily limited. Hence, the maintenance of at least small primary forest areas within agricultural landscapes is essential for the sustainable conservation of epiphyte diversity.
P0097 – ePoster
Climate change, medicinal plants and ethnobotany in
Nepal Himalaya
Kunwar, RM
RW 4
1 , Karki, JB 2 , Shrestha, KP 3 , Bussmann,
3
1 Ethnobotanical Society of Nepal, Kathmandu; 2 Dept of
National Parks and Wildlife Conservation, Kathmandu;
Natural History Museum, Tribhuvan University,
Kathmandu; 4 William L Brown Centre, Missouri
Botanical Garden, USA
Climate Change, Medicinal Plants and Ethnobotany in
Nepal Himalaya Abstract: Nepal has witnessed accelerated glacier melting, deglaciation, rainfall variation, impacts on forests and biodiversity, landslides, alien species invasion, etc. due to climate change. It is hypothesized that the forest ecosystem is severely affected with altering forest types, species population, association, regeneration, etc. due to climate change since changed in plant species distribution, richness, association, phenology, behavior, etc. were pronounce in
Nepal Himalaya. Impact of climate change on indigenous medicinal plants is intense and it is complemented by invasion of alien species. Furthermore, these indigenous medicinal plants are not immune to climate change and are at more risk due to the finite limits at the tops. These plants are more stressed producing abnormal secondary metabolites in a warmer environment as albedo changes are likely to have higher mortality, dieback and vary on phenology, and quality and quantity of the products, and the plants may no longer own their therapeutic potential.
All these changes have been known to affect harvesters’ and cultivators’ abilities to grow, collect and use the species. It is therefore argued that there is a need for more research into the effects of climate fluctuations on indigenous medicinal plants and ethnobotanical information.
P0098 – ePoster
Community signatures of phenotypic integration in foliar traits
Lechowicz, M 1 , Gilbert, B 2 , Radovski, M 1
2
1 McGill University, Montreal, Quebec, Canada;
University of Toronto, Canada

Photosynthetic capacity, leaf mass per area and foliar nitrogen concentration are key elements in an integrated set of foliar traits that underpin the function of leaves as photosynthetic organs. Although these traits vary widely among species, this variation falls along a leaf economic spectrum (LES) organized by broadly consistent interrelationships that indicate a high degree of functional integration (Wright et al. 2004; Shipley et al. 2006).
Depending on their evolutionary adaptation, species occupy different positions along the axes of the LES. At any point along the spectrum, the foliar characteristics of a given species fall within a constrained range of variation around what appears to be a global trend in the alternative designs that have emerged in the course of vascular plant evolution. In a particular environmental regime, species with somewhat different combinations of traits can achieve comparable levels of foliar function in conjunction with variation in other elements of functional design at the whole-plant level (Marks & Lechowicz
2006). In turn, as a plant community assembles in a locality, only those species in the regional species pool that have foliar traits consistent with the LES for the environmental regime in that locality can enter the community. The dominance-diversity relationships in the community should be determined, at least in part, by the degree to which the foliar traits for each species in the community are, or are not, at the global optima defined by the LES in that environmental regime. If this line of reasoning is valid then we can make two predictions.
First, the bivariate relationships between traits in the LES should be stronger when expressed as community-wide averages weighted by species abundance across a series of plots than when expressed, as is usually the case, by the average trait values of species in the plots. This convergence toward the global trend in LES relationships reflects both the role of plasticity in individual plants and the nature of community assembly at the scale of neighboring plants (Messier et al. 2010). Second, some bivariate relationships between foliar traits may differ for communities comprising distinct functional groups. This reflects the fact that bivariate variation around global trends in the LES can arise in evolutionarily and functionally distinct designs that organize the tradeoffs among foliar and other traits at the whole-plant level in fundamentally different ways to achieve comparable performance in a given environmental regime. We show support for these predictions in comparisons of communities of ferns and juvenile trees growing in a series of 50 m 2 understory plots in an old-growth, temperate deciduous forest.
P0099 – ePoster
1
Distribution patterns of
Hyparrhenia hirta
(L) Stapf
(C
4
),
Merxmuellera disticha
(Nees) Conert (C
Themeda triandra
Forsskal (C neighbour interactions on the Signal Hill, Cape Town,
South Africa
Ligavha-Mbelengwa, M 1 , Bond, W 2
4
3
) and
) and nearest
, Bhat, R 1
2
University of Venda, Limpopo Province, South Africa;
University of Cape Town, South Africa
The present investigation describes the competitive interactions between three perennial grasses,
Hyparrhenia hirta (L) Stapf (C
4
), Merxmuellera disticha
(Nees) Conert (C
3
) and Themeda triandra Forsskal (C
4
) which coexist on the east-and west-facing slopes of
Signal Hill, Cape Town. Through quadrat sampling it was determined which of those species was most dominant relative to the other two species. Competitive interactions were inferred from pattern analysis. In the first place, the distribution patterns of the three grasses were analysed using a dispersion index (R). In all five cases, except one, distributions tended towards regularity; the exception tended towards aggregation.
Secondly, nearest-neighbour analysis was used to detect inter-and intra-specific competition. Ratios of the diameters of the smaller individuals to the distances from their neighbours showed that for individuals of the same size, both inter- and intra- specific nearest-neighbours were located at nearly the same distances. The slopes of regression lines of the combined cover versus distance for different nearest-neighbour pairs were also compared.
Results indicated the relative intensity of competition among and within species. M. disticha appeared to be growing better than H. hirta , whilst T. triandra , wherever it occurred with M. disticha , appeared to be competitively stronger. As a result regular distribution is anticipated for the species in the long-run. In some cases, intraspecific competition was relatively more intense than interspecific, whilst in others interspecific was rather more than intraspecific. Accordingly both mixed and segregated populations are feasible, as well as the exclusion of weak competitors.
P0100 – ePoster
The ecological role of fog and foliar water uptake in three woody species from southeastern Brazilian
1 cloud forests
Lima, AL 1 , Oliveira, RS 1
Institute of Biology, State University of Campinas
(UNICAMP), Brazil
Fog is an important water source for plants in cloud forests but future climate scenarios predict that the cloud base in tropical mountains will move hundreds of meters upwards, exposing plants to extreme water deficits.
There is scarce information about the mechanisms by which cloud forest species use fog water, and how dependent they are on this water source for survival. The objective of this study was to demonstrate that direct uptake of fog water by the foliage is a common mechanism of water acquisition for cloud forest species.
We also evaluated the anatomical pathways involved in the process of foliar water uptake and the role of fog deposition on the ecophysiological performance, growth and survival on three common woody species from
Brazilian cloud forests: Drimys brasiliensis , Eremanthus erythropappus and Myrsine umbellata . Greenhouse experiments using artificial fog enriched in deuterium and labeling experiments with fluorescent apoplastic tracers demonstrated that all species absorbed water directly through their leaf surfaces. Leaf water uptake contributed to as much as 42% of total foliar water content in D. brasiliensis . Water diffused directly through leaf cuticles in all species. Tector trichomes of E. erythropappus and glandular peltate trichomes of M. umbellata are important pathways for water uptake in these species. After the cuticular barrier was transposed,
391

solutions moved through apoplastic routes at the epidermis and parenchyma in all species, reaching the xylem in E. erythropappus and accumulating on collector cells of M. umbellata . The high abundance of hydrophilic compounds found on leaf surfaces of D.
brasiliensis and
M. umbellata may have contributed to the great retention of solution with apoplastic tracers in the mesophyll of these species. Sprayed water on isolated branches caused an immediate increase in leaf water status in all species, a benefit also observed in a long-term experiment. After two months, plants under the fog treatment improved leaf water potential, photosynthesis and stomatal conductance in relation to plants exposed to drought in the greenhouse experiment. Species responses to the treatments were different. D. brasiliensis had 100% mortality after a month under drought. Soil water content (SWC) of fogged plants was constant throughout the experiment, suggesting that fog water might have been used as the main water source for transpiration during this period. E. erytroppappus was the only species with similar growth and survival rates in all treatments and the beneficial effects of fog water was only observed after two months.
For M. umbellata , SWC decreased substantially in the drought and fog treatments but no decrease in water potential, photosynthesis and stomatal conductance was observed in the fog treatment. For this species, fog water deposition caused a decoupling of foliar water relations and gas exchange from soil water availability. We demonstrated that the three cloud forest woody species can absorb water directly through their leaves during fog deposition, allowing shoot rehydration and improving ecophysiological performance, growth and survival when soils are dry.
P0101 – ePoster
China grasses: phylogenetic influence on biogeographical pattern and ecological trait evolution of C
4
photosynthetic subtypes
Liu, Q 1 , Peterson, P
Sciences;
2 , Ge, X 1
1 South China Botanical Garden, Chinese Academy of
2 Dept of Botany, National Museum of Natural
History, Smithsonian Institution, Washington DC, USA
We study the influence of phylogeny on biogeographical pattern and relations of climatic factors to evolution of C
4 photosynthetic subtypes for China grasses. The diversity database of 145 grass species was collected from 115 nature reserves in China, their climatic factors (mean annual temperature and precipitation) and photosynthetic pathways (C
3
and three C
4
subtypes) and plastid DNA sequences (ndhF, rbcL and matK) were collected simultaneously. The phylogenetic influence on biogeographical pattern of grasses was analysized in
Phylocom. The rates of climatic niche evolution across the phylogenetic tree were analysized in Brownie.
Furthermore, the relations of climatic factors to evolution of C
4
photosynthetic subtypes were analysized in SPSS.
Within PACCMAD clade, the multiple rates of temperature niche evolution was marked in the C
3
/C
4 node AP, the multiple rates of precipitation niche evolution was marked in the C
3
/C
4 node AX according to both AIC difference and chi-squared P-value. Among boxplot analysis, the divergence width of climatic niches in C
3
nodes were the widest, whereas the divergence
392 width of climatic niches in C
3
/C
4 insignificantly different. Among C
4
nodes were
nodes, the divergence width of temperature niches of NAD-ME nodes were wider than NADP-ME nodes, the divergence width of precipitation niches in NADP-ME nodes were wider than NAD-ME nodes, a novel interpretation for the ecological sorting of C
4
subtypes was provided. The results revealed that phylogenetic characteristics in different lineages played a critical role for distribution patterns of two major clades along temperature gradient and two major subfamilies along precipitation gradient in
China.
P0102 – ePoster
In the line of fire: deciphering why some grasses respond to smoke-derived germination-stimulants whilst others do not
Long, R 1,2 , Griffiths, E
Merritt, D 1 , Powles, S 1
2 , Adamek, M 2 , Stevens, J 2 ,
1 School of Plant Biology, University of Western
Australia, Crawley, Australia;
Garden, West Perth, Australia
2 Kings Park and Botanic
The regeneration of plants from seeds following fire can be mediated by both physical and chemical effects.
Chemically speaking, there are compounds in smoke and ash that can stimulate germination, such as the karrikins, of which karrikinolide (KAR1) is the most naturally abundant. Whilst KAR1 commonly triggers seeds in the
Brassicaceae, Solanaceae and Asteraceae families to germinate, species in the Poaceae – another major understory and weed family – have afforded mixed results. This study aimed to understand why some grass species respond to KAR1 whilst others do not, by testing the response of known KAR-responsive and nonresponsive grass species to KAR1 during dormancy alleviation under natural and laboratory conditions.
Using a field-based seed-burial trial, dose-response experiments, and stratification experiments, we investigated whether KAR1 could alleviate dormancy and trigger seeds to germinate for three global agronomic weeds: Avena fatua , Lolium rigidum and Eragrostis curvula . Seeds of Avena fatua were consistently stimulated to germinate with KAR1 in all experiments, whether seeds were freshly-collected or dormancy had been partially alleviated. In contrast, seeds of L. rigidum and E. curvula failed to respond to KAR1 when the seeds were fresh, after-ripened in the laboratory, and even during natural dormancy loss in the field. Interestingly, although KAR1 did not stimulate germination in L. rigidum and E. curvula , it did alleviate dormancy when applied during stratification, indicating that KAR1 can alleviate dormancy in some species and trigger germination in others. Moreover, the finding that species that were previously thought to be non-responsive to
KAR1 can be manipulated to respond to it, suggests that the molecular mechanisms for karrikin responses may exist in all species, but may not be realised under natural conditions. These findings are helpful for understanding and predicting the responses of grass species following fire. They also contribute to a growing body of research aimed at using karrikinolide as a tool for triggering uniform germination of seeds of both fire-responsive and

non-fire-responsive species, for enhancing restoration efforts and depleting the weed seed bank.
P0103 – Poster
Hot subantartic megaherbs: thermal effects of leaf and flower gigantism
Lord, J 1 , Little, L 1 , Tomlinson, V 1 , Huggins, L 2
1 Botany Dept, University of Otago, New Zealand; of Conservation, Invercargill, New Zealand
2 Dept
Temperature can drastically limit growth in cold climates and for biotically pollinated plants, low temperatures can also limit pollinator availability. Alpine plants have evolved a range of features that are thought to enable survival in cold climates. These include dense or reflective leaf hairs and compact growth forms. The climate of the World Heritage Subantartic Islands, unlike most alpine zones, is consistently wet, cold and overcast.
With less than 650 sunshine hours per year and frequent gale-force winds, even 'summer' air temperatures seldom exceed 10 o C. The endemic megaherbs found on these islands are remarkable for their massive leaves and inflorescences and also for their unusually coloured flowers. It has been speculated that leaf and flower gigantism in Subantartic megaherbs provides a thermal benefit to growth and pollination in the cold, overcast conditions typical of Subantarctic islands. We tested this hypothesis for five megaherb species over a two week period on Campbell Island (latitude 52 o South) using an
Fluke infra-red thermal imaging camera. Leaf temperatures markedly higher than ambient were only observed in species such as Pleurophyllum speciosum
(Astercaeae) and Stilbocarpa polaris (Araliaceae) which have massive, hairy, corrugated leaves. These species showed pronounced rapid heating up to 15 o C above ambient temperatures during even brief periods of increased solar radiation, despite significant and consistent wind chill. Species with smooth, but still large leaves, showed little or no heating effects suggesting that the modification of the leaf boundary layer is a significant factor in the observed thermal effects.
Significant increases in flower temperature were observed for a range of species, which, with the exception of the dioecious Bulbinella rossii , all possessed very dark reddish-purple flower pigments. In the massive yellow flowers of Bulbinella rossii , heating was maximal in male inflorescences, particular in the zone of young flowers were nectar content was highest and pollen was being freshly released. Pollinator activity
(mainly small Diptera ) was largely restricted to this zone suggested that the combination of heat and rewards was a very effective attractant. These findings provide a starting point for unravelling the evolution of these striking plants, in particular how they may have adapted to survive on these inaccessible and inhospitable southern islands.
P0104 – ePoster
Anthropogenic impacts on the Brazilian Atlantic
Forest: genetic effects on the threatened tree
Dalbergia nigra
(Leguminosae)
Resende, LC 1 , Ribeiro, RA 2 , Lovato, MB 1
1 Universidade Federal de Minas Gerais, Belo Horizonte,
Brazil; 2 Universidade Federal dos Vales do
Jequitinhonha e Mucuri, Diamantina, Brazil
The Brazilian Atlantic Forest, one of the richest biomes in the world, has been intensively deforested and fragmented due to different types of land use.
Considering that species endemic to this biome are restricted to forest fragments, they may be more sensitive to the genetic effects of habitat fragmentation. Therefore, we aimed at studying the genetic diversity and population structure of the threatened species Dalbergia nigra , a tree endemic to the Brazilian Atlantic Forest, in a recently fragmented area. A total of 119 adults and 116 saplings were sampled from five populations. Two were located in a large reserve, one of them in a well preserved area of primary forest, and the other three in fragments of different sizes and degrees of disturbance. All samples were genotyped for six microsatellite loci. The well preserved population showed the highest levels of genetic diversity, which were significantly higher than the diversity of saplings from impacted populations.
Traditional means of measuring population structure was significant and ranged from low to moderate (pairwise
FST = 0.028-0.152), while the recently developed Jost’s
D was considerably higher (pairwise DEST = 0.039-
0.301). However, genetic structure among saplings was lower than among adults (FST = 0.056; DEST = 0.231 for saplings versus FST = 0.088; DEST = 0.275 for adults), suggesting an increase in gene flow after fragmentation that could partly buffer the loss of genetic diversity following fragmentation and logging. Despite recent fragmentation, we found evidence of genetic diversity loss in the most disturbed populations.
Considering the presence of private alleles in impacted populations and the suggestion of increased gene flow, it is important to preserve the extant fragments in order to maintain diversity and avoid long-term effects of inbreeding and drift. This study was supported by CNPq
(Conselho Nacional de Desenvolvimento Tecnológico,
Brazil) and FAPEMIG (Fundação de Amparo à Pesquisa do Estado de Minas Gerais, Brazil).
P0105 – ePoster
Biological and ecological peculiarities of
Heracleum sosnowskyi
distribution in Russia
Luginina, E 1
1 Russian Research Institute of Game Management and
Fur Farming, Russia
Invasive species are considered to be one of the threats to biodiversity and economy. In connection with increasing mobility of human, quick development of transport, tourism and world trade, biological invasions can become the base of ecological disintegration due to expanding spread of alien species. And to work out complex measures intended for monitoring, localization and extirpation of alien species we need to study their biological and ecological peculiarities. Such investigations are especially urgent in case of Heracleum sosnowskyi which appeared in taiga zone of European
Russia in late 40’s as highly productive fodder plant for livestock. H. sosnowskyi (Apiaceae (Unbelliferae)) is new adventive species for taiga flora of Russia which
393

distribution reflects common trends of synanthropic changes of flora. It is south-moderate European species native to Caucasus, Transcaucasia, Asia Minor where it inhabits forest edges and subalpine meadows. As a result of spreading from cultural lands the species occupied different regions of the country and their number is rapidly increasing. During the period of 2000-2009 the areas of H. sosnowskyi stands near large settlements increased more than 3 times. During past 18 years (1991-
2009) H. sosnowskyi marked at first as solitary plants has formed solid stands along highways, penetrated into meadows, bushes and forest edges. According to some estimations the areas of H. sosnowskyi stands on agricultural lands increased 2 times during 2005-2008. H. sosnowskyi is a tall (up to 3.5-4 m high) biennialtriennial plant. All age groups are presented in the population. The age spectrum is left-side. Juvenile plants prevail in coenopopulations (51%), immature individuals
– 20%, generative – 17%, senile – 12%. The density of mature individuals reached 4+0.1 per m 2 178;. Seed reproduction is basic for H. sosnowskyi and the study of seed productivity revealed that the seed potential of 1 generative shoot is about 6 thousand seeds. Hence each square meter of a stand can produce 24000 seeds.
Germination ability in lab conditions is about 62% and
56% in field. If to take field germination ability then 1 m 2 of H. sosnowskyi coenopopulation can provide more than
13 thous. seedlings occupying approx. 2 hectares. High seed productivity and germination ability ensure intensive seed renewal of the stand. Reproduction potential of the species is best fulfilled in synanthropic habitats but there are many cases of forming of H. sosnowskyi populations in undisturbed areas. Populations of H. sosnowskyi in most of Russian districts are stable, self-reproducing without repeated introduction.
Intensively forcing out aboriginal species H. sosnowskyi breaks normal functioning of local ecosystems and causes the decrease of biodiversity. To prevent uncontrolled distribution of alien species in natural habitats the Ministry of Natural Resources suggests development and realization of a system which includes revelation of basic transit paths, inventory and monitoring of alien species, prognostication and estimation of risks of potential invasions. As for today we only have methodical recommendations of how to control the distribution of H. sosnowskyi and also
European manual which both offer temporary measures.
P0106 – ePoster
Flowering dynamics and pollination of
Rhynchospora ciliata
(Vahl) Kükenth (Cyperaceae): ambophily as a strategy to maximize reproductive success
Costa, ACG 1 , Machado, IC 1
1 Departamento de Botânica, Universidade Federal de
Pernambuco – UFPE, Recife, Brazil
Cyperaceae is characteristically a wind pollinated family, although readaptation to entomophily has already been reported for some representatives, including
Rhynchospora ciliata . In this study we aim to elucidate:
1) is R. ciliata pollinated by bees, wind or both vectors?
2) which floral traits indicate anemophily and/or entomophily in R. ciliata ? 3) the biotic pollen vectors increases the reproductive success in this species? The
394 field observations were performed from August 2007 to
June 2010, at the Dois Irmãos Ecological Reserve, a remnant of Atlantic forest located in the urban area of
Recife, northeastern Brazil. The reproductive phenology was monitored, as well as the floral biology, the frequency and behavior of insects on flowers. We conducted experiments to test which parts of flowers are more attractive to the visitors and whether the pollination is also mediated by wind. The reproductive system was analyzed based on the results of spontaneous selfpollination and natural pollination (control), manual selfing and outcrossing. Rhynchospora ciliata has spikelets with hermaphrodite (bisexual) flowers at the base of the inflorescence which have three stamens, superior ovary and a style with a bifid stigma. The neutral (morphologically staminate) flowers are located at the apex of the inflorescence, and are composed of three staminodes and often a pistillode. We can recognize four anthesis stages in bisexual flowers, distinguished by the development of stamens, style and ovary. During anthesis the filaments elongates exposing the anthers. At the same time the style also grows up allowing the contact with visitors and facilitating the pollen capture through the air, by the stigma. Each floral scape lasts about two months and flowering is continuous throughout the year. We recorded four species of bees pollinating R. ciliata contacting the ventral part of its abdomen with pollen, only floral resource, and with stigmas. The flowers are characteristically anemophilous, however the white floral bracts could be considered an entomophilous trait. We found through the tests of floral attractiveness that the absence of exposed anthers interferes with the average number of insect visits, while the presence or absence of white floral bracts is indifferent to the pollinators, indicating possible memorization of the flowers to be visited. Reproductive tests show that R. ciliata is self-incompatible and that ambophily can be considered a strategy that increases fruit set in this species.
P0107 – ePoster
The backbone of oil flower networks: the importance of plant and bee species in different biomes
Mello, MAR 2 , Bezerra, ELS 1 , Machado, IC 1
1 Departamento de Botânica, Universidade Federal de
Pernambuco – UFPE, Recife, Brazil; 2 Institute für
Experimentelle Ökologie, Universität Ulm. Albert-
Einstein, Germany
Interactions between oil-collecting bees and oilproducing flowers are a very specialized mutualism with strong mutual dependence. At community level, those interactions have been seldom studied. Evidence from the single network study available suggests that species with more interactions are more important for maintaining the whole structure of oil flower networks. Therefore, we aimed at investigating the hierarchy of species in oil flower networks and its biological basis. Byrsonima oil flowers and Centris and Epicharis oil bees, which are the best adapted for oil interactions, were expected to form the backbone of those systems: a subset of highly important species that are either hubs (i.e. have many interactions) or connectors (i.e. bind together parts of the network). As floras and faunas vary geographically, we

also expected the importance of each species to vary among biomes. By compiling data from 40 papers on oil interactions, we built six networks for different Brazilian biomes, from steppes to rainforests. Although 90% of the species played peripheral roles, a few were pointed out as very important. On average, plants played more important roles than bees. Oil flowers of the genera
Byrsonima and Banisteriopsis were hubs or connectors in most networks, and oil bees of the genera Centris and
Epicharis were connectors in most biomes. Our findings suggest that species with a longer coevolutionary history, which are well adapted for oil interactions, form the backbone of the highly specialized oil flower networks.
Moreover, the importance of those central species varies among biomes, suggesting that additionally to adaptation, local availability of partners is an important structuring factor. Our study also shows how different the concepts of specialization in networks and ecological theories are from each other, leading to opposite classifications for the same species.
P0109 – ePoster
Ethnobotanical evaluation of some angiosperms from
Aravalli Ranges
Mahmoudi Otaghvari, A 1
1 University of Mazandaran, Mazandaran Province of
Iran
In human society, plants play an important role since time immemorial. The study of relationship between plants and human beings in term of benefits are called
Ethnobotany. Angiosperms are by far the most numerous divers and successful extant plant group containing well over 95% of all land plant species alive today. Flowering plants grow in virtually every habitable region and are dominant in some aquatic and most terrestrial ecosystems. Angiosperms comprise the great bulk of our economically important plants including our most valuable food crops. In present electronic world modern man's life still depends fundamentally on plants. The three basic necessities of life food, shelter and clothing and a host of essentials are still derived mainly from the plant- kingdom. The Aravalli is oldest mountain range in the world. Aravalli ranges have wide range of habitats, varied climate, physiographic landscape, rocks, soil types and geological antiquities. There is no independent systematics account of angiosperms of Aravalli range.
The present paper deals with some of important valuable angiosperms, which are found in the Aravalli range. Key words: Ethnobotany, Angiosperms, Aravalli range.
P0111 – ePoster
Lichens on man-made substrata in Russian cities: what substrate will be next?
Malysheva, N 1
1 Komarov Botanical Institute RAS, Russia
Lichens are not indifferent to the substrate. This is confirmed by the methods of mathematical statistics and is used in the determination. A special group is formed by lichens on man-made substrata. It is interesting from a scientific and practical point of view. The approaches to the classification of that lichen group could be different.
It can be physical and chemical properties of the substrate, the utilitarian use of objects, the typical habitats. We used all these approaches. Our and herbarium collections, literature data were analyzed.
Lichens on man-made substrata were studied in 110
Russian cities. The particular attention was given to lichens found on building materials. There were identified 123 species of lichens. On the processed stone were found 27 species. On the treated wood were identified 78 species. On ceramic materials, which included bricks, were registered 21 species. Inorganic binders were populated with lichens worse. On the cement were found 12 species, on slate – only 3.
Concretes and mortars were populated by lichens better.
29 lichens were met on concrete, 23 species – on reinforced concrete. Metals were colonized by lichens also poorly. 9 species were found on iron roofs. Thermal insulation materials were colonized by lichens poorly as well, there were found 10 species. On the waterproofing materials were obtained few lichens, only 3 species. On polymeric building materials were recognized few species (2), but sometimes they formed a large coverage.
The lichen group registered on building materials contains about 20 % species identified in the Russian cities. This demonstrates the sufficiency of their activity.
We have found 2 species on flax rope and cardboard cover also. The taxonomic structure is quite similar in different cities. The most frequent genera were
Caloplaca , Candelariella , Lecanora , Phaeophyscia ,
Physcia , Xanthoria . Crustose lichens dominated (58 % of total), less were foliose (32 %) and little - fruticose (10
%). There were mostly widespread species. They occur in different elements of the urban landscape – in the parks, cemeteries, residential buildings, etc. Cultivated plants are a special substrate for lichens. Among the cultivated plants are varieties and hybrids, which are not found in nature, such as x Cerapadus fontanesiana . On this plant we have collected lichens Lecanora hagenii , L. symmicta , Xanthoria polycarpa . We have found 60 lichen species on 26 kinds of fruit trees and shrubs in the northwest of Russia. New problems arise in the study of lichens on man-made substrata. There are the study of lichens on new synthetic materials (plastic, etc.) and genetically modified plants. Time and our future research will show what kind of the substrata will be the next.
P0113 – Poster
How much genetic variation is stored in the seed
1 bank?
Mandak, B 1 , Zakravsky, P 1 , Mahelka, V 1 , Plackova, I
Institute of Botany, Pruhonice, Czech Republic
1
The idea that seed bank might have evolutionary consequences was first formulated by Templeton and
Levin (1979). In their view, the seed bank can sometimes serve as an important reservoir of genetic variation, buffering the effects of local extinction of genotypes in adult populations caused by selection or drift. However, no empirical studies have investigated the genetic relationship between above- and belowground populations on larger data set than one species over maximally five localities. Hence, almost no empirical
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studies have investigated the genetic relationship between seed banks and surface plants across larger spatial scales. These larger scale, multipopulation studies are necessary to test whether there are broad genetic differences between soil seeds and surface plants, rather than merely idiosyncratic differences between these lifehistory stages in particular populations. We have tested hypothesis concerning seed bank genetic diversity using three species with different life history cycle to improve our understanding of whether and to what extent soil seed banks affect the genetic structure of entire surface plant populations.
P0114 – ePoster
Floristic composition of a fragment of Atlantic Forest in São Paulo, Brazil
Martins, E 1 , Cruz, A 1 , Santos, A 1
1 CEPEMA-USP, São Paulo, Brazil
The study was conducted in an ecological park Pereque, located at coordinates 23°15'05''S and 51°01'40''W, in the city of Cubatao, Sao Paulo, in the conservation area of atlantic forest. Represents an important conservation area boundaries to do with the Parque Estadual Serra do Mar
– Núcleo Itutinga-Pilões and by presenting the remaining primary and secondary vegetation of Montana Rain
Forest and Lower Montane dampening the anthropogenic pressure coming from the industrial pole of Cubatao. The study aims to survey the floristic composition of the park and contribute to the preparation of management plan for the same. Fortnightly collections were made of specimens of vascular plants in the period 2009 and 2010 by random walks on trails and in the interior of the fragments. In this preliminary floristic survey were reported until now a list of 121 species belonging to 31 families and 86 genera, where seven species are ferns.
The families with the greatest number of species were
Poaceae and Fabaceae (11), Asteraceae, Rubiaceae and
Myrtaceae (9), Lauraceae and Euphorbiaceae (7),
Cyperaceae (5), Solanaceae, Melastomataceae,
Urticaceae (4) and Piperaceae and Meliaceae (3). The genera with most species were Cyperus (Cyperaceae), with 5 species, Piper (Piperaceae) with 4, Miconia
(Melastomataceae), Panicum (Poaceae) and Solanum
(Solanaceae) at 3. Altogether, Trichilia silvatica C.DC.
(Meliaceae) and Brosimum glaziovii Taub. (Moraceae) were present in the list of species considered endangered by IUCN (International Union for Conservation of
Nature). The study found a diversity index of Shannon
(H') for species of 3.01 nats. individual -1 , a reasonable rate compared to other surveys in the Atlantic Forest in
Sao Paulo. This species diversity associated with the presence of endangered species suggests the importance that the park is to conserve the biodiversity of the
Atlantic Forest and the need to prepare a management plan aimed at planning and adequate control of their public use for recreational activities.
P0115 – ePoster
Conservation genetics of the endemic root holoparasite,
Dactylanthus taylorii
McLay, T 1
Symonds, V 1
, Tate, J 1 , Holzapfel, A 2 , Gemmill, C 3 ,
1 Massey University, New Zealand;
Conservation; 3
2 Dept
University of Waikato, New Zealand of
Identification of intraspecific genetic variation is important for management and conservation of endangered taxa. Dactylanthus taylorii
(Balanophoraceae) is an endemic New Zealand root holoparasite on approximately 30 native trees and is endangered due to threats from introduced pests, particularly brush-tailed possums, and habitat degradation. Dactylanthus taylorii is listed as nationally vulnerable and management is coordinated through a
National Recovery Group, who actively monitor and protect populations throughout its North Island range.
For this study, we are using microsatellite loci to evaluate genetic diversity within and between populations of D. taylorii to determine population structure and to aid in conservation management decisions. To isolate microsatellite loci, a random sample of genomic DNA was sequenced using 454 GS FLX. From the 61,709 random sequences that were generated, 4044 repeats were identified and 753 primer pairs were designed using
MSATCOMMANDER and PRIMER3. Based on criteria related to sequence quality and repeat structure, 48 loci have been screened to identify consistently amplifiable and variable markers that will be suitable to evaluate the genetic structure of all known populations. Fifteen microsatellite markers will be used to determine the population structure of genetic variation, and the level of inbreeding within the species. This information will directly benefit future conservation efforts as the
Department of Conservation and the National Recovery
Group determine populations of national significance to conserve and potential seed sources for translocation to increase genetic diversity in existing populations and to establish new populations. These results will provide new insight into the conservation and management of parasitic plants, especially those restricted to small, declining populations in isolated habitats.
P0116 – ePoster
Structuring of genetic diversity in three endangered wild daffodils endemic to mountains of southeastern
Iberian Peninsula,
Narcissus bugei
(Fern Casas),
N. longispathus
Pugsley and
N. nevadensis
Pugsley
Medrano, M 1 , Herrera, CM 1
1 Estación Biológica de Doñana, CSIC, Seville, Spain
Biodiversity hotspots provide particularly favourable places for the study of the evolutionary mechanisms underlying the appearance of new species. The Baetic
Ranges, in the southern Iberian Peninsula, is one of the most remarkable biodiversity hotspots in the
Mediterranean Basin, and is characterized by the high endemism level of many plant lineages. One of the latter involves species of the Mediterranean genus Narcissus .
DNA variation was studied at the intra- and interspecific levels in three endangered species of this genus endemic to mountains of the southeastern Iberian Peninsula, N . bugei , N. longispathus and N. nevadensis (section
Pseudonarcissi). The taxonomic status of these three
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closely related taxa is still under debate, which most probably reflects ongoing evolutionary processes.
Interspecific differences in geographical ranges and habitat specialization are expected to yield contrasting patterns of population genetic structure. Whereas N. longispathus and N. nevadensis had very narrow distribution ranges, and are strict habitat specialists confined to permanently water-logged meadows, a very rare and isolated habitat in the Mediterranean region, N. bugei has a wider distribution range and it can grow in a broader variety of Mediterranean habitat types, such as sclerophyllous forests, deciduous shrublands, or poorly drained meadows, suggesting a more recent history of population isolation. Amplified fragment length polymorphisms (AFLP) fingerprints were generated for a total of 679 individuals belonging to 36 populations of these three endemic taxa of Narcissus . Sampled populations encompassed the whole distribution ranges of the three taxa. We used four primer combinations, which generated a total of 577 scorable fragments per genotype. Preliminary results revealed that patterns and magnitude of genetic divergence among populations are strongly correlated with geographical distribution.
Results are consistent with predictions that (i) specieslevel, overall genetic diversity was directly related to geographical range size, and (ii) despite their narrower distribution ranges and higher geographical isolation of their populations, N.
longispathus and N. nevadensis , also have high genetic structure at small spatial scales.
High genetic structuring exhibited by these narrow endemics across the Baetic Ranges is compatible with the hypothesis that enhanced genetic diversification of lineages can partly account for high plant diversity levels in this biodiversity hotspot.
P0117 – ePoster
Anatomy of the vegetative organs of 17 Neotropical species of Gentianaceae Juss. as a subside to taxonomy and phylogeny
Dalvi, V 1
A 1
, Meira, R 1 , Coutinho, I 1 , Silva, L
1 Universidade Federal de Viçosa, Brazil
1 , Azevedo,
The Gentianaceae family has around 1,650 species, 87 genera, and 6 tribes (Chironieae, Exaceae, Gentianeae,
Helieae, Potalieae and Saccifolieae), besides the genus
Voyria , without a definite position. Such classification considers phylogenetic affinities based on molecular data which demonstrate congruence with the external morphology, palynology, and chemical taxonomy. This family shows great anatomical and morphological diversity. Circumscriptions of the species and genera have not been well established yet. This study aims to describe the anatomy of the vegetative organs of species belonging to genus Calolisianthus (Helieae), Curtia and
Hockinia (Saccifolieae), and Deianira (Chironieae); to contribute to the knowledge of the family; and to select useful anatomical characters to taxonomy and phylogeny of this group. Specimens of Calolisianthus pedunculatus ,
Deianira damazioi , Curtia tenella , Curtia tenuifolia ,
Curtia patula and Curtia diffusa were collected at different Rupestrian Fields (Rock Outcrops) in the State of Minas Gerais, Brazil. Samples of Curtia obtusifolia ,
Curtia conferta , Curtia verticillaris and Hockinia montana from voucher material were also analysed as well as specimens of Declieuxia fruticosa and Declieuxia cordigera , which were the Rubiaceae species used as the external group in the cladistic analysis. Samples of leaves, stems, and roots were processed according to the usual plant anatomy methodology for epidermal dissociation, cleared preparations, methacrylate and paraffin embedding, histochemical tests, and scanning electron microscopy. For the cladistic analysis, data from literature of seven species belonging to the tribes studied were included. Altogether, 59 anatomical characters were analysed. The Maximum Parsimony Analysis was run by the Paup 4.0.b10 version program. All the species structurally analysed showed anomocytic stomata placed at the same level of the other epidermal cells, nectarioles, colleters, and stems with stomata and internal phloem.
The characters considered as diagnostic for the different species are as follows: type of mesophyll; type of vascular bundle; distribution and type of stomata; presence of hypodermis; presence of cuticular flanges; outlining of the epidermal cells; presence of crystals; patterns of venation; shape of the stem; number of wings in the stem; presence of aerenchyma, mycorrhiza and
Casparian strips evident in the roots; type of root growth; and variation on the type and distribution of the colleters and nectarioles. According to the cladistic analysis,
Gentianaceae is monophyletic a bootstrap value of 100%.
Three monophyletic clades giving support to the
Calolisianthus (67% value of ‘bootstrap’), Curtia (89%) and Hockinia genus (86%) also emerged. On the other hand, Deianira showed polytomies probably due to insufficient anatomical data on the species belonging to this genus, especially data regarding the secretory structures. Data on the secretory structures, pattern of venation, and type of stomata showed to be promising to the phylogeny of this group. It should also be highlighted the importance of using anatomical characters to solve taxonomic problems concerning the different species of
Gentianaceae. The necessity of data from different sources of characters in order to achieve a more accurate analysis of the phylogenetic relationships among the different taxonomic categories of Gentianaceae should also be emphasized. We thank CNPq and FAPEMIG for the financial support.
P0118 – ePoster
Influence of soil physical properties on plant functional ecology of the Mussununga ecosystem,
Brazil
Saporetti-Junior, A 1
M 1 , Araujo, D 2
, Schaefer, C 1
, Meira-Neto, J 1
, Souza, A 1 , Soares,
1 Universidade Federal de Vicosa – Botany Postgraduate
Program, Brazil; 2 Universidade Federal do Rio de
Janeiro, Brazil
Distribution ranges of plant species are related to physical variables of ecosystems that limit plant growth.
Therefore, each plant species response to physical factors builds up the functional diversity of an ecosystem. The higher the species richness of an ecosystem, the larger the probability of maintenance of functions and the higher the potential number of Plant Functional Types
(PFT.) Thus, the richness potentially increases the number of functions of the highly diverse Atlantic
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Rainforest domain in Brazil. On the other hand, severe plant growth limitations caused by stresses decrease species richness. In Spodosols of Mussununga, an associated ecosystem of Atlantic Rainforest, the percentage of fine sand is directly related to water retention. Moreover, the depth of cementation layer in
Mussununga’s sandy soil is a physical factor causing flooding stress for plants, and where it is added to low water retention in soils with low fine sand percentage it causes the double stresses of flooding in rainy season and water scarcity in dry season. This study aimed to identify plant functional types (PFTs) among Mussununga plant species responding to water stress gradients of soil and to verify the effects of the gradients on plant species richness of Mussununga. Canonical Correspondence
Analysis (CCA) of species abundance and soil texture variables was performed on 18 plots in six physiognomies of Mussununga. Species richness rarefactions were calculated for each vegetation form for comparisons of diversity. The two main axes of CCA showed two PFTs responding to soil texture and cementation layer depth: stress tolerator species and mesic species. Diversity of plants is affected by physical variables. The species richness rises as fine sand proportion also rises in Mussununga. The effect of the cementation layer is not significantly related to species richness variation.
P0119 – ePoster
First insight into microbal community diversity associated with
Sphagnum denticulatum
(Sphagnaceae)
Melosik, I 1 , Turska, A 1
1 Adam Mickiewicz University, Pozna ń
, Poland
Background: To learn more about the anonymous consortium of organisms (algae, bacteria, fungi, nematodes) associated with gametophores of Sphagnum denticulatum and preserved epiphytically and in intercellular spaces between living chlorophyllous cells
(i.e. within hyaline cells), we surveyed for sequence similarities among the DNA sequences obtained from a
Sphagnum denticulatum random-sequence DNA library.
Information reported here may yield deeper insight about the composition and structure of the Sphagnum denticulatum community. Methods: The samples of
Sphagnum denticulatum gametophores were taken directly from field populations at two locations in Poland
(shorelines of Krasne Lake, 53°22´00´´N; 17°16´40´´E, and P³oczyca Lake 54°05´02´´N, 17°25´01´´E). Total nucleic acids were extracted as described by Schlink &
Reski (2002). DNA was digested with FastDigest EcoRI and established in the plasmid pBluescript II KS (+). A total of 285 randomly chosen clones from the DNA library were sequenced and analyzed using BLASTn,
BLASTx, and tBLASTx with low complexity filter
(NCBI GenBank, http://www.ncbi.nlm.nih.gov). The query size ranged from 500-1800bp. Results: The information obtained from sequence similarity indicate that gametophores of Sphagnum denticulatum harbor mainly acid-tolerant, aerobe or anaerobe, Gram-negative, nitrogen-fixing diazotrophic members of plantassociating
α
- and ß-subgroups of Proteobacteria.
Bacteria that inhabit ecosystems such as water or sediments from fresh water lakes were also detected among
α
-, ß-, and Deltaproteobacteria. The other groups of the library were determined to be in the Acidobacteria, whereas Cyanobacteria, although detected, show low sequence similarity. Of the 273 good quality sequences,
19% belongs to these microbial groups. However, there was no clone whose fragment of sequence was 100% identical of known bacteria species, i. e. sequence similarity ranged from 70% to 83%; the sequence overlap ranged from (74) 85% to 100%. It has been repeatedly reported that under the conditions were nitrogen is limiting, as in wetlands, the N is provided by natural atmospheric N deposition and by N-fixation via
Cyanobacteria (blue-green algae), e.g. Nostoc sp. living epiphytically or intracellularly in peat mosses. Our finding suggests rather low cyanobacterial diversity (and low sequence similarity), even in plants collected from two separate locations. The results indicate that bacterial community associated with Sphagnum may be composed of unknown Proteobacteria species. Although, a definition of bacterial species based upon 16S-23S rRNA gene sequencing still is not fully established, 16S rRNA gene has emerged as a preferred genetic region for bacteria species-specific identification, therefore additional studies are required.
P0120 – ePoster
Comparative movements of sympatric common and rare wasp pollinators and implications for pollen flow
1 in sexually deceptive orchids
Menz, MHM 1, 2 , Dixon, KW 3 , Peakall, R 3
2
Kings Park and Botanic Garden, West Perth, Australia;
School of Plant Biology, University of Western
Australia, Crawley, Australia; 3 Research School of
Biology, Australian National University, Canberra,
Australia
Conservation of specific pollination relationships, such as those observed in sexually deceptive orchids requires an understanding of the ecology of both the plant and the pollinator. The movement capability of the pollinator has significant implications on gene flow of the plant.
Pollination by sexual deception may confer the advantage of long-distance pollen flow, as males are hypothesized to leave the vicinity of an orchid following attempted copulation. Males of Zaspilothynnus gilesi and
Z.
nigripes (Hymenoptera: Thynnidae) are the sole pollinators of the orchid species Drakaea elastica and D. livida respectively. D. elastica is Critically Endangered and endemic to the Swan Coastal Plain, Western
Australia, while D. livida is relatively common and widespread in the south-west of Western Australia. We tested whether common and rare species showed similar movement patterns in continuous habitat to determine the potential connectivity of patchily distributed orchid populations. Movement patterns of Z. gilesi were compared with those of Z. nigripes using a markrecapture study. Male wasps were attracted by baiting with picked orchids. In addition to recapturing individuals using orchids, patrolling males were also captured opportunistically by sweep-netting. Recapture distances ranged from 0-556 m and 0-267 m for Z. gilesi and Z. nigripes respectively. For Z. gilesi , recapture distances while baiting ranged from 0-300 m, median =
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38 m (n = 132). Z. nigripes recaptured while baiting moved from 0-150 m, median = 46.5 m (n = 58). The maximum distance moved by Z. gilesi is up to 5 times longer than that previously reported for Thynnid wasps, while that of Z. nigripes is up to twice as long. These results are likely to be conservative in terms of the maximum distance these species can move. This indicates that in continuous habitat, patchy populations of these Drakaea species are likely to remain connected via pollen flow. As to how these wasp species respond to fragmented habitat requires further research. While conducting this study, it was observed that D.
elastica flowers from the northern part of the species range did not attract Z. gilesi , as the southern orchids do. Rather, northern D. elastica attracted a closely related species of wasp distinguished by morphology. Reciprocal baiting experiments have shown that northern orchids do not attract the southern wasps, and northern wasps do not alight on the southern orchids. This means that these northern populations of D. elastica need to be managed separately. Baiting across 42 areas of suitable habitat has demonstrated that this northern pollinator appears to be rare (1.5% of sites), suggesting that further investigation into the ecology of this wasp species is warranted.
P0121 – ePoster
Ex situ conservation of seeds of native species of
Georgian flora at the Caucasus Regional Seed Bank,
Georgia, and the Millennium Seed Bank of the Royal
1
Botanic Gardens, Kew
Mikatadze-Pantsulaia, T
L 1 , Kikodze, D 1
Trivedi, C 2
, Barblishvili, T
, Terry, J 2
1 , Eristavi, M
, Sutcliffe, V 2
1
1 , Kobakhidze,
, Khutsishvili, M 1 ,
Tbilisi Botanical Garden and Institute of Botany, Tbilisi,
Georgia; 2 Royal Botanic Gardens, Kew, UK
Georgia is located at the heart of one of 34 'biodiversity hotspots', where conservation works are especially urgent. Ex situ conservation via establishing seed bank and living collections provides an effective insurance against loss of biodiversity in the wild. Of special importance is conservation of endemic, rare and endangered species of economically important plants.
Our work is aimed at ex situ conservation of seeds of endemic and rare species of Georgian flora at the
Caucasus Regional Seed Bank (CRSB) in Georgia. The work is the result of on-going collaboration between the
Department of Plant Conservation of the Tbilisi
Botanical Garden and Institute of Botany, and the
Millennium Seed Bank (MSB), Royal Botanic Gardens,
Kew. As a result of joint activities accomplished within the frame work of the Millennium Seed Bank Project the seed bank now holds seed of 848 rare and endangered plant species. This encompasses nearly 21% of Georgian flora. Out of all collections, 183 are endemic to the
Caucasus and 65 are Georgian endemics. The secured species belong to 97 families (52% of the total number of families recorded in Georgia) and 389 genera (38% of the total number of genera recorded in Georgia). Thirtyfive species in the collection are relic species. Many are also crop wild relatives. The collected seeds are deposited at the CRSB, and the duplicates are sent to the
MSB. Each seed collection is accompanied by a herbarium specimen. For each seed collection made a data form is completed with information on locality, ecological and habitat data, population size, species distribution, threats, taxonomic information, plant description, vernacular names, ethnobotanical data, and collectors' information. After processing, all data is transferred to the CRSB electronic database using
BRAHMS (Botanical Research and Herbarium
Management System) and shared with the MSB. Seed treatment and deposition corresponds to modern standards accepted at the MSB. Prior to deposition in cold storage, seeds are germination tested in petri dishes using either filter paper or agar. Any seed remaining at the end of a germination test are checked by cut tests under a light microscope. The germination percentage of each seed collection is calculated as a measure of the viability of the collection. Seedlings produced as a result of germination tests are planted into pots and thus living collections of around 60 rare species have been established. Similarly, horticulturists at the Royal
Botanic Gardens, Kew, have established living collections from some Georgian seed collections.
Collected seeds are dried in an incubator-drier or using silica-gel. The moisture status of a seed collection is measured using a Rotronic hygrometer. For storage at low temperature (-20 o C), the moisture status of a collection is reduced to 15-20% equilibrium Relative
Humidity. Once seeds have reached the appropriate moisture status, they are placed in foil bags and sealed.
The seeds are stored at -20 o C at the Caucasus Regional
Seed Bank and the accompanying herbarium specimens are held at the Georgian National Herbarium (TBI).
Duplicate seed collections and specimens are sent to the
Millennium Seed Bank.
P0122 – ePoster
The reproductive capacity and ex situ conservation of some species of genus
Campanula
of primary conservation concern for Georgia
Mikatadze-Pantsulaia, T
Gabedava, L 1
1 , Jgenti, L 1 , Melia, N 1 ,
1 Tbilisi Botanical Garden and Institute of Botany, Tbilisi,
Georgia
Genus Campanula is represented in Georgia by 66 species, of which 28 are Georgian endemics and 16 are endemics of the Caucasus. Reproductive capacity of 3
Georgian endemic species of genus Campanula -
Campanula kachetica Kantsch., C. raddeana Trautv. and
C. kemulariae has been studied for the first time in places of occurrence of their native populations. All three species are subject to protection as highly ornamental economic species of extremely localized distribution.
The present state, fertility and viability of individual populations were evaluated, exact locations of populations were recorded by means of Global
Positioning System. With the aim to establish selfreproduction capacity of a species structural aspects of seed formation were investigated. Fertility of female and male generative spheres has been established.
Reproductive capacity – the ratio of a number of actually formed seeds (actual seed producing capacity) to a number of ovules set (potential seed producing capacity) was calculated for each target species. Terms of seed ripening and collection were established. Germination
399

capacity of seeds, sprouting and development of seedlings were tested experimentally in laboratory conditions on petri dishes, open ground and pots.
Favorable conditions for seed germination – temperature, illumination regime have been established. In contrast to literary data preferential importance of high temperature for germination of seeds of target species against cold stratification has been revealed. All collections of seeds of the studied species germinated rapidly and have shown high germination percentage. Germination percentage was high for C. kachetica and C. kemulariae – 95.6% and 91.4% correspondingly. Collection of seeds of C. raddeana revealed low germination percentage – 40%.
Ex situ conservation activities have been undertaken for target species: seed bank and collection of seedlings sprouted from seeds were established. The living collections of species under study have been created by means of rapid vegetative propagation by rhizomes. As a result of division/partition of rhizomes average of 5-10 individuals have been obtained from a single plant.
P0125 – ePoster
Shading effects on the reproductive ecology of
Besseya Bullii
, a rare species
Chi, K 1 , Molano-Flores, B 2
Seoud, D 1
1 University of Illinois, USA;
Survey, USA
, Collins, M 1 , Abou-El-
2 Illinois Natural History
Besseya bullii (Plantaginaceae) is a rare, prairie-savanna plant species endemic to the Midwestern United States of
America. During the 1980s, this species was a candidate for U.S. federal protection as a threatened species, but ultimately lost consideration due to insufficient knowledge of its biology. Earlier surveys of this species listed woody encroachment as a primary conservation concern because it can limit resources (i.e., light, space, nutrients and water) for B. bullii . In this study, we wanted to determine the effects of shading on reproduction of B. bullii populations in the state of
Illinois (USA); specifically, we were interested in the effects of shading on inflorescence morphology (e.g., flower density), reproductive output (e.g., fruit/seed set), and fitness (i.e., seed germination). Populations were visited in 2008 (n = 8) and 2009 (n = 10), and assigned to one of three habitat categories according to degree of canopy closure: open, semi-shaded, and shaded. At each population, 20 infructescences were randomly collected and used to determine: (1) fruit density (i.e., number of fruits per centimeter), (2) fruit set (i.e., proportion of fruit produced from total flowers), (3) seed set (i.e., proportion of seeds produced from total ovules), and (4) germination
(i.e., proportion of seeds germinated from a sample of
100 seeds randomly selected from infructescences per population). Results from infructescences sampled in
2009 show that fruit density was significantly higher for open and semi-shaded habitats compared to shaded habitats. Moreover, fruit and seed set for open and semishaded habitats were significantly higher than for shaded habitats in both years. Lastly, seed germination did not differ among habitats in either year of the study. These results show that closed canopy habitats negatively impact reproduction in B. bullii by reducing fruit density and fruit/seed set. Shading modifies photosynthesis rates,
400 and thus affects the development of reproductive organs and offspring. Furthermore, studies suggest that shading alters the foraging behavior of insect pollinators, which may further impede fruit and seed production in shaded populations. Based on our findings, we recommend woody species removal to promote and increase the reproductive success of B. bullii .
P0126 – ePoster
Reproductive success in gynodioecious
Lobelia
1 spicata
: effects of population size and sex ratio
Ruffatto, D 1 , Molano-Flores, B 2
University of Illinois, USA; 2 Illinois Natural
History Survey, USA
Gynodioecy, or the classification assigned to all joint hermaphrodite and female plant breeding systems, has been documented in only 7% of angiosperms. In this system, female plants must depend on hermaphroditic pollen donation since they are unable to produce pollen, and may experience lower pollinator visitation due to their lack of nectar production and smaller flower size. It has been suggested that female plants may compensate for these deficiencies by producing more flowers, larger fruit and seed sets, and larger, more quickly-germinating seeds than their hermaphroditic counterparts. The objective of this study was to determine if the predicted patterns of gender differences between hermaphrodite and female plants are found for Lobelia spicata
( Campanula ceae), a gynodioecious prairie species.
Lobelia spicata is a short-lived, herbaceous perennial that is common in loess, sandy and loam prairies throughout eastern North America. Its terminal raceme inflorescence matures acropetally, with flowering and fruiting occurring from May to August. Flowers are zygomorphic ranging in color from white to blue with blue to white nectar guides, respectively. This species is pollinated primarily by small bees, the most common being members of the Augochlorella genus. During summer
2008 and 2009, eleven L. spicata populations located across Illinois (USA) were visited to gather the following measurements for both plant genders: population counts, gender ratios, fruit set, seed set, seed biomass and percent seed germination. High variation in population size and gender ratio was observed among the surveyed populations. Additionally, differences in female and hermaphrodite fruit sets, seed sets, seed biomass and seed germination rates were found. Gender patterns appear to be similar to other gynodioecious systems, although variation among populations was also found suggesting that the means by which females are maintained varies among populations (e.g., pollinator visitation, mycorrhizal mutualism dynamics).
P0127 – ePoster
The ant foraging on extrafloral nectaries of
Ipomoea
1 pes-caprae
(Convolvulaceae) in the dune vegetation: ants as potential antiherbivore agents
Mondal, AK 1 , Mondal (Parui), S 2
Vidyasagar University, India; 2 Lady Brabourne College
(Under Calcutta University), India

Among the different members of the dune vegetation which help in the formation and stabilization of sand dunes, Ipomoea pes-caprae has acted as a principal sand binder along the sandy shores and nearby areas of Bengal and Orissa, India. But unfortunately this species is gradually disappearing from this belt due to the rapid increase in agriculture, housing, recreation and tourist resorts and for this, effective management policies for the conservation and protection of this species is necessary.
But fortunately enough this species has its own defence mechanism to protect itself from being overgrazed. It is due to the presence of extrafloral nectaries. Extrafloral nectar is an indirect, generally ant-mediated, defence mechanism that is particularly common in tropical plants.
This study focuses on the different groups of arthropods visiting and the interactions among these different groups of arthropods visiting extrafloral nectaries of Ipomoea pes-caprae. The diurnal activity patterns of arthropods on nectaries were recorded. This paper also reports the presence of similar extrafloral nectaries in other species of Ipomoea.
P0128 – ePoster
Genetic basis of local adaptation in
Arabidopsis halleri ssp. gemmifera
: spatial and temporal variations in functional genes
Morinaga, S 1 , Nagano, AJ 2 , Ito, M 1
1 University of Tokyo, Japan; 2 Kyoto University, Japan
Adaptation to the local environment is the major driving force in evolution. Especially for plant species, gradients in environmental characters cause adaptation to different habitats, resulting in ecotypic differentiations as well as speciation. Although the genetic basis of local adaptation is one of the major topics in evolutionary biology, it is difficult to conduct such genetic studies on most wild species due to the lack of knowledge about their genome.
Arabidopsis halleri ssp. gemmifera is most closely related species to A. thaliana and it demonstrates phenotype differentiation attributed to altitudinal gradients. Two ecotypes grow in the lowland and highland habitats on Mt. Ibuki in Japan, providing a favored study system for elucidating the mechanisms of local adaptation along altitude. To uncover genetic basis of local adaptation, we conducted cross-species genomic microarray analysis in two ecotypes of A. halleri ssp. gemmifera using AtMap1, tilling array for A. thaliana . In these results, we found eighteen genes whose signal intensities significantly differentiated between lowland and highland types. These genes encoded several metabolic enzymes, disease resistance proteins, kinese family proteins, and a transcription factor. Furthermore, we analyzed the spatial and temporal variations in those functional genes using voucher specimen sampled from herbarium. In this presentation, we will discuss the functions of these genes for different environmental factors, such as temperature and wind.
P0129 – ePoster
Interactions between dark root endophytes and
Subterranean Clover (
Trifolium subterraneum
L.) in soil
1
Mukasa M, Tendo T 1 , McGee, P 1
University of Sydney, Australia
, Saleeba, J 1
Facultative fungal endophytes have been widely documented in above-ground plant tissues, where the endophyte-host plant interaction ranges from mutualism to antagonism. Documentation of non-mycorrhizal fungal endophyte colonisation of roots is limited. A collection of endophytic fungi isolated from 406 plant roots across the Sydney basin (NSW, Australia) resulted in the isolation of more than 900 endophytes, 120 of which were melanised. Sterile soil sown with germinated seed of Trifolium subterraneum L. was separately inoculated with each melanised fungus and plant growth responses recorded. Plant growth over 7 weeks followed three broad patterns: (1) The majority of endophytes neither enhanced nor inhibited plant growth compared to the uninoculated control; (2) A second large group initially inhibited seedling growth, but seedlings recovered and mostly were similar in size to the control after 7 weeks;
(3) A very small number were seedling pathogens, inhibiting growth. The melanised isolates most likely contain dark septate endophytes (DSE), a diverse group of root-inhabiting fungi whose actions within host plants include growth promotion. These fungi are being further investigated for their potential influence on soil aggregation and carbon sequestration within soil, factors beneficial to plant growth.
P0130 – ePoster
Does Proteaceae cluster-root activity benefit phosphorus uptake and growth of neighboring
1 shrubs?
Muler, AL 1 , Oliveira, R 1 , Lambers, H 2 , Veneklaas, E 2
Universidade Estadual de Campinas (UNICAMP), São
Paulo, Brazil;
Australia
2 University of Western Australia, Sydney,
Species that inhabit phosphorus(P)-impoverished soils often have typical adaptations for enhancing P uptake, such as cluster-roots. However, there are several species that co-occur in the same environment that lack such clear adaptations. The aim of this study was to investigate whether two of these species ( Scholtzia involucrata and Hibbertia subvaginata ) can benefit from the cluster-root P mobilization by a co-occurring Banksia species. We conducted two greenhouse experiments: the first one used a split-root design where in one half of the pot roots of Scholtzia or Hibbertia intermingled with one side of the Banksia attenuata root system, while in the second one we used a replacement series design, where
Banksia and Scholtzia shared a pot at proportions of 2:0,
1:2 and 0:4. We found that where the amount of clusterroots was greater, the [P] in young leaves of Scholtzias was higher and the total biomass of Hibbertias was greater. However, Banksia formed less cluster-roots and regular roots in the side of the pot with the neighbour plant. Phosphorus concentration was also higher in the young leaves of Banksia when plants grew with another
Banksia than when they grew with two individuals of
Scholtzia , despite the smaller size of Scholtzia s.
Additionally, plants of Scholtzia grew more next to one individual of Banksia and one individual of Scholtzia
401

than with three other individuals of Scholtzia . Our results demonstrate that Banksia can positively influence the growth and foliar [P] of neighbouring plants. However, the facilitative effects of cluster roots can be offset by the apparent ability of Banksia to sense their neighbours and produce most of their cluster-roots far away from them.
P0131 – ePoster
Pollination network from a mangrove in northeastern
Brazil: The importance of flies (
Diptera
) as pollen vectors
Nadia, TL 1 , Mello, MAR de 2 , Machado, IC 3
1 Centro Acadêmico de Vitória, Universidade Federal de
2 Pernambuco, Vitória de Santo Antão, Brazil; Institute für Experimentelle Ökologie, Universität Ulm, Germany;
3 Departamento de Botânica, Universidade Federal de
Pernambuco, Recife, Brazil
The mangrove is a tropical ecosystem from intertidal areas and it is characterized by having few plant species.
This ecosystem has been threatened by the increase in human population is coastal areas. It is, therefore, important to investigate not only its species, but also the interactions among them. Network theory has been very helpful to study interactions at community level, allowing us to understand how they affect the structure and fragility of the ecosystem services delivered. So, our objective was to analyze a plant-pollinator interaction network from a mangrove community, in order to understand its structure and so provide subsidies for its conservation. Data collection was performed from June
2002 to May 2006 in a mangrove in northeastern Brazil.
There are only four plant species in the studied area:
Rhizophora mangle (Rhizophoraceae), Avicennia schaueriana (Acanthaceae), Conocarpus erectus and
Laguncularia racemosa (Combretaceae). As the first is anemophilous, we analyzed only the last three.
Pollinators were collected, identified and assessed in terms of frequency, behavior and result of the visit
(pollination or nectar robbing). We analyzed only the pollinators. The following metrics were calculated: network size, number of observed interactions, connectance and number of interactions per species
(average degree). The degree of nestedness was calculated with the NODF index. Dependence among species and relative interaction strengths were calculated from the frequency of visits. We recorded 37 pollinator species, resulting in a total of 40 species in the network.
We observed 61 interactions, resulting in a connectance of 0.55. The average degree was higher for plants (20.3) than form animals (1.65). 49% of the animals interacted with only one plant species. The degree of nestedness
(0.53) observed was not significant. Only four species of flies and a wasp interacted with the three mangrove species, the other 14 pollinators (bees, butterflies, flies and wasps) interacted with two mangrove species and 18 with only one. The fly Palpada albifrons showed the highest relative strength (0.55). 50% of the pollinators showed high dependence on A.
schaueriana , 35% on C. erectus and 15% on L. racemosa . Thus, A. schaueriana had the highest relative strength (0.51) among plant species. High connectance and low degree of nestedness are expected in networks with few species. Since nested networks are more robust to extinctions, pollination
402 networks of mangrove may be more fragile to disturbances than other pollination networks; this might have serious consequences, considering that this ecosystem is under severe threat. Although the network was not nested, it was possible identify a core of highly connected species, formed by four species of flies, mainly P. albifrons , and a wasp, unlike other communities, where the core is formed mainly by bees.
Therefore, flies play an important role in the mangrove pollination network, being important for the maintenance of mangrove plants and the animal species that depend on them.
P0132 – ePoster
Relationship between flowering intensity of
1
Melocactus
species (Cactaceae) and hummingbird’s flower visit
Nadia, TL 1 , Machado, IC 2
Centro Acadêmico de Vitória, Universidade Federal de
2
Pernambuco, Vitória de Santo Antão, Brasil;
Departamento de Botânica, Universidade Federal de
Pernambuco, Recife, Brasil
Melocactus is a genus of Cactaceae widely distributed in
Caatinga vegetation, a dry forest in the semiarid region, northeastern Brazil. Their representatives are pollinated by hummingbirds and cases of hybridization in this genus are known. In this work, we aim to analyze the relationships among climatic factor, flowering intensity and frequency of visit, throughout the year, among three sympatric Melocactus species and their hummingbird pollinators. Phenological data were collected monthly from November 2009 to January 2011 in two localities of
Parque Nacional do Catimbau, Canyon (8°31’55.40”S and 37°15’3.08”W, 914 m alt.) and Serrinha
(8°30’59.24”S and 37°14’2.23”W, 914 m alt.), northeastern Brazil. In the first locality, nine individuals of Melocactus violaceus and four of M. zehntneri were tagged, while in the second locality, we marked 23 and
21 individuals of M. zehntneri and M. ernestii , respectively. The frequency of visits was monthly calculated on the number of times a hummingbird foraging for nectar. The number of visited flowers per visit bout was also recorded. These data were collected from January 2010 to January 2011. We used the historical average rainfall to run the analyses. The rainy season occurs from January to August (>60 mm), dry season from September to December (<50 mm), and average annual rainfall is 1098 mm. Pearson correlation coefficient was calculated among the rainfall, flowering intensity and frequency of visit pair-wise. Melocactus violaceus and M. ernestii flowered continuously, while
M. zehntneri did not flower during four months (Apr-Jul) in Canyon and flowered continuously in Serrinha. The flowering peaks occurred in Nov/2009 and Dec/2010 in
Canyon and in February and September in Serrinha. Only
M. zehntneri flowering was negatively correlated with rainfall at both sites. Two hummingbird species were recorded in every locality, Chlorostilbon aureoventris at both sites, Anopetia gounellei only in Canyon and
Eupetomena macroura only in Serrinha. Chlorostilbon aureoventris was recorded in almost months, except in
July at both sites showing the highest frequency in
February, October and Jan/2011 in Canyon and in June

and Jan/2011 in Serrinha. Anopetia gounellei was not recorded in Fev-Mar and Jun-Jul, showing the highest frequency in Jan/2010 and in August, while E. macroura was the less frequent hummingbird, being recorded only in five months. The number of visited flower/visit of each species was correlated with frequency of visit.
There was no relation between the frequency of visit of any hummingbird and rainfall or flowering intensity.
However some features may be directly correlated with visited flowers/visit. The flowering intensity of M. zehntneri (Canyon) and M. ernestii (Serrinha) and the number of flowers of focal plants (both sites) were correlated with the number of visited flowers by C. aureoventris (both sites) and A. gounellei (Canyon).
Eupetomena macroura may not present any correlation because it is probably a non-resident hummingbird.
Therefore M. zehntneri and M. ernestii seem to be important to the attraction and retention of residents hummingbirds in their respective locality (Canyon and
Serrinha), while M. violaceus is crucial in the months which M. zehntneri is not in bloom.
P0133 – ePoster
Pollination ecology and reproductive biology of
Chrysobalanus icaco
L. (Chrysobalanaceae) in coastal vegetation, northeastern Brazil
Barral, EL 1 , Nadia, TL 2 , Machado, IC 3 , Lopes, AV 3
1 Programa de Pós-Graduação em Biologia Vegetal,
2
Universidade Federal de Pernambuco, Recife, Brasil;
Centro Acadêmico de Vitória, Universidade Federal de
3
Pernambuco, Vitória de Santo Antão, Brasil;
Departamento de Botânica, Universidade Federal de
Pernambuco, Recife, Brasil
Chrysobalanaceae is a pantropical family of about 500 species, with distribution center in the Neotropics. The genus Chrysobalanus consists of only three species in the world, among which two species and one subspecies occur in Brazil ( Chrysobalanus icaco , C. venezuelanus and C. icaco subsp. icaco ); Chrysobalanus icaco is typical of coastal vegetation. The family as a whole is poorly studied for reproductive systems and pollination.
Therefore, in view of the importance of studies in this area for the maintenance and conservation of species, ecosystems and considering that coastal vegetations have suffered high levels of degradation, the aim of this study was to examine the floral biology, the pollination systems and the reproduction of C. icaco in Restinga vegetation in Pernambuco, Brazil (7º33’ S e 35º00’ W).
Monthly phenological observations were made to register the intensity of buds, flowers and fruits. The species showed continuous flowering and fruiting with peaks in the dry season. Flowers are actinomorphic, pentamerous, arranged in cimose inflorescences, hermaphrodite, and offer nectar as its primary resource. Anthesis starts around 7:00 pm and the flowers are completely opened at
9:00; ca. 16:00 they began to wilt. Pollen/ovule ratio and pollen viability was high, indicating that C. icaco seems to present facultative xenogamy. Chrysobalanus icaco is generalist regarding its pollination system, being pollinated by diverse small insects, as flies, bees, wasps, butterflies, and diurnal moths. However, the wasps were considered the main pollinators due to its higher frequency of visits (ca. 80% of the visits). With respect to the reproductive system, the species is self-compatible, setting fruits after manual geitonogamy, manual crosspollination, and natural pollination. Thus, C. icaco depends on biotic vectors for pollination, however, the generalist pollination system can ensure sexual reproduction of the species.
P0134 – ePoster
Oil pollution effects on the mangroves
Avicennia marina
and
Bruguiera gymnorrhiza
Naidoo, G 1 , Naidoo, Y 1
1 University of KwaZulu-Natal, South Africa
The effects of Bunker fuel oil on morphological and physiological responses of Avicennia marina and
Bruguiera gymnorrhiza were investigated in glasshouse and field experiments. In the glasshouse study, 15month-old seedlings of A. marina were subjected to oiling or debarking treatments for 6 months. Oiling or debarking of a 5 cm ring of the basal portion of the stem, alone and in combination, reduced leaf CO
2
exchange by over 50% and resulted in the production of adventitious roots immediately above the debarked and/or oiled stem
8-12 weeks after the commencement of treatments. In the field study, sediment oiling at a single dose of 5L m-2 of
A. marina and B. gymnorrhiza trees reduced electron transport rate (ETR) through Photosystem II (PSII) and
PSII quantum yield. Oiling also reduced the photochemical efficiency of PSII (Fv/Fm) in B. gymnorrhiza , but not in A. marina . After 15 weeks of oiling, adventitious roots developed at the base of the stem in A. marina , but not in B. gymnorrhiza . Naturally occurring A. marina seedlings with adventitious roots exhibited lower leaf CO
2
exchange rates, photochemical efficiency of PSII and leaf chlorophyll content than similar seedlings without these roots. These results indicate that Bunker fuel oil adversely affects photosynthetic performance of A. marina and B. gymnorrhiza mangroves. Avicennia marina responds to oiling by producing adventitious roots at the base of the stem. Adventitious root production at the base of the stem may be a useful biological indicator of oil or other toxic pollutants in A. marina .
P0135 – ePoster
Variation of floral morphology in
Aster hispidus var. tubulosus
(Asteraceae): correlation between
1 frequency of floral types and microhabitat
Nakagawa, S 1 , Ito, M 1
Dept of General Systems Studies, Graduate School of
Arts and Sciences (Biology), University of Tokyo, Japan
Variation in floral morphology would have direct effect to speciation. Thus, its origin and maintenance mechanism seem to be important for diversification of plants. Family Asteraceae shares a composite flower head, comprising ray and/or disc florets. They have ligulate and tubular florets which are known to be regulated by CYCLOIDEA like gene. Aster hispidus var. hispidus has ligulate ray florets in the outermost wheel,
403

as is typical in Asteroideae. However, this is not the case with A. hispidus var. tubulosus , which is characterized by long tubular ray florets. Moreover, these ray florets show extreme variation within a population. In this study, we surveyed floral variation of A. hispidus var. tubulosus in the population alongside an armlet of Tenryu River,
Nagano pref., central Japan. The study population was subdivided into small patches, which differed in the frequency of each floral type. We found that the frequency of flowers with long tubular ray florets is positively correlated with the lightness of the patches.
We will discuss this result in relation to possible difference in pollinator attractiveness due to morphological variation in ray florets. In addition, to clarity genetic mechanism of the floral variation, we determined sequences ortholog of CYCLOIDEA gene in
A. hispidus var. tubulosus , and will report association between floral types and their sequences.
P0136 – ePoster
Limnological study of Cyanophyceae Of Unai thermal spring of Gujrat (India)
Nandan, S 1
1 Dr PR Ghogrey Science College, Dhule, India
A thermal spring is a spring which is produced by the emergence of geo thermally heated ground water from the earths crust. Many workers have paid their attention towards certain thermal springs from different parts of
India. They studied thermal springs for taxonomical aspects of algae and recorded many algal taxa belonging to class Cyanophyceae, Bacillariophyceae and
Chlorophyceae. But in Gujarat State of India very few workers have paid attention to thermal springs. In the present study Unai thermal spring of Navasari district of
Gujarat was selected for limnological and biodiversity studies of Cyanophyceae for the period of six months.
The collection of water samples & algal samples were made from 4 different stations of Unai thermal spring.
The physico-chemical analysis of water of Unai thermal spring has carried out from November 2006 to April
2007. In present study the biodiversity of Cyanophyceae group of algae was studied. Total 14 genera and 47 spcies of Cyanophyceae group includes taxa belonging the orders Chroococcales and Nostacales. The dominant genera of Cyanophyceae, viz. Microcystis , Chroococcus ,
Gleocapsa , Gleotheceae , Aphanocapsa , Aphanotheceae ,
Synechococcus , Merismopedia , Dactylococcopsis ,
Arthrospira , Spirulina , Oscillatoria , Phormidium &
Lyngbya, were recorded from 4 stations of Unai thermal spring. The most dominant genera, viz. Chrococcus (8),
Gleocapsa (8) & Oscillatoria (8), were observed in group of Cyanophyceae Oscillatoria laetevirens ,
Phormidium africanum , Phormidium orientale ,
Aphanocapsa thermals are definately thermal forms whereas other forms recorded in the present study were found in cold water as it is tallied with earlier findings.
P0138 – ePoster
Implications of clonal integration on water use of
Hydrocotyle bonariensis
subjected to contrasting water conditions in a coastal dune of Ubatuba, São
Paulo, Brazil
404
Nery Cardoso, I 1 , Vieira Santello, G 1
Negri Bernardino, P 1
Oliveira, R 2
, Brasci B, G 1
, Angeluzi Jardim, V 1
,
, Silva
1 State University of Campinas – UNICAMP, São Paulo,
Brazil; 2 Dept of Plant Biology – UNICAMP, São Paulo,
Brazil
Sandy soils in coastal dunes are restrictive habitats for the establishment and growth of plants because they are characterized by low nutrient and water availability and high salinity levels. Such habitats are dominated by plants with stoloniferous growth, which may allow the transfer of resources among ramets from favorable to unfavorable sites. The objective of this study was to evaluate the role of clonal integration in ecophysiological performance of Hydrocotyle bonariensis . We tested the hypothesis that connected ramets would have higher short-term fitness when subjected to water deficits than fragmented ramets . This study was conducted in a sanddune area in the southeastern coast of Brazil, Ubatuba –
São Paulo. To evaluate the short-term fitness of the local plants, we measured stomatal conductance and transpiration rates of twelve individuals of H. bonariensis subjected to the following treatments: integrated and fragmented ramets (irrigated and nonirrigated with sea water). In each treatment, we considered parental-ramets (further from the ocean) and offspring-ramets (closer to the ocean). The irrigation simulated a high tide reaching the offspring plants. The salty water induced stomatal closure in all offspring ramets, but the magnitude of the response varied among the treatments. Fragmented offspring-ramets that received salty water reduced stomatal conductance and transpiration rates in comparison to integrated ramets, indicating that the water stress of ramets close to the sea can be relieved by the translocation of water from wellhydrated parental-ramets. Clonal integration allows the transference of resources available from ramets in favorable microhabitats to ramets in less favorable sites, and can explain the success of plants with stoloniferous growth on such stressful environments.
P0139 – ePoster
Invasive dinoflagellate
Ceratium furcoides
in two tropical reservoirs
Nishimura, PY 1 , Moschini-Carlos, V 2 , Pompêo, MLM 1
2
1 Universidade de São Paulo, Instituto de Biociências,
Departamento de Ecologia, São Paulo, Brazil;
Universidade Estadual Paulista Júlio de Mesquita
Filho, Departamento de Engenharia Ambiental, Campus
Experimental de Sorocaba, São Paulo, Brazil
Ceratium is a large freshwater mixotrophic dinoflagellate protected by a rigid cellulose armor. Although Ceratium is a characteristic summer inhabitant of temperate stratified lakes with low surface nutrient concentrations, since 1999, reports of high densities of Ceratium in tropical and subtropical eutrophic waters became more frequent, such as Argentina, Chile, South Africa, New
Zealand and Australia. Since 2007, Ceratium is frequently found in Brazilian eutrophic reservoirs. We found the species Ceratium furcoides (Levander)
Langhans, 1925 in phytoplankton samples from two

reservoirs in São Paulo city, Brazil: Billings
(Taquacetuba branch) and Guarapiranga (Parelheiros branch), in September 2009. Billings reservoir is used for electric power generation, leisure, fishery and navigation.
Guarapiranga’s main use is water supply. According to the State Environmental Agency, the main problem of both reservoirs is the excess of organic matter from clandestine domestic sewage input. Because of that,
Billings is mainly eutrophic and Guarapiranga, mesotrophic. Consequently, phytoplankton blooms, especially cyanobacteria, are frequent in both reservoirs.
We found C. furcoides in all 18 samples from Billings reservoir and only in four out of 18 samples from
Guarapiranga reservoir. In Billings reservoir, the density ranged from 11 to 297 cells/mL, while in Guarapiranga reservoir, ranged from 15 to 30 cells/mL. C. furcoides was first recorded in Billings reservoir in 2008. The authors suggested that the appearance of C. furcoides caused the reduction of cyanobacteria density. In our study, we found low cyanobacteria density along with C. furcoides population. Further studies are required to explore C. furcoides -Cyanobateria interaction in detail.
Here, we report the first occurrence of C. furcoides in
Guarapiranga reservoir. Since 2000, water from Billings reservoir (Taquacetuba branch) is pumped to
Guarapiranga reservoir (Parelheiros branch) during the dry season, when the water level of the last is low.
Probably, C. furcoides was transferred to Guarapiranga reservoir during this pumping. The lower frequency and density of C. furcoides observed in Guarapiranga reservoir, suggest that the colonization of C. furcoides in
Guarapiranga reservoir is still in early stages comparing with the colonization in Billings reservoir. Further studies are required to investigate: (1) the interaction of this dinoflagellate with the phytoplankton and zooplankton community; (2) how this species is being dispersed; and (3) what are the consequences of the C. furcoides colonization for the water supply.
P0140 – ePoster
Spatial heterogeneity of aquatic macrophytes related with the trophic gradient in urban tropical reservoir
Rodrigues, MEF 1
Nishimura, PY 3
, Macedo, CCL 2
, Pompêo, MLM 3
, Hirata, RT 3 ,
1 Universidade de São Paulo, Instituto de Biociências,
2
Departamento de Botânica, São Paulo, Brazil;
Universidade Estadual Paulista Júlio de Mesquita
Filho, Departamento de Engenharia Ambiental, Campus
Experimental de Sorocaba, São Paulo, Brazil;
3 Universidade de São Paulo, Instituto de Biociências,
Departamento de Ecologia, São Paulo, Brazil
Aquatic macrophytes varies considerably in their morphology, physiology, life cycle and adaptive strategies that enable them to tolerate different kinds of stresses such as the variation of water level, light, availability of nutrient and temperature. The aim of this study was to investigate the composition and distribution of aquatic macrophytes in Guarapiranga reservoir (Sao
Paulo, Brazil), which owns 70% of its perimeter in the largest city of South America, São Paulo, featuring it as an urban reservoir. Guarapiranga reservoir has an area of
3618 ha, with total volume of 194.000.000.000 L and the average depth of 7 m. It supplies the southwestern part of
São Paulo with potable water, which covers more than 3 million people. The reservoir also functions as flood control, power generation and recreational activities.
Previous studies revealed the heterogeneity of the reservoir, comprising oligotrophic to eutrophic regions.
Field samples were collected monthly from June/2008 to
May/2009, in 20 macrophyte stands distributed throughout the reservoir. The similarity between the macrophyte stands was calculated from presence/absence matrices, using the grouping analisys of the cluster type, with the similarity measure of Sørensen and connection method type UPGMA. Aquatic macrophyte community of the reservoir was composed of 145 species belonging to 93 genera distributed in 45 families, being 139 angiosperms, 5 ferns and 1 bryophyte. The families with the greatest number of species were Cyperaceae (33 species), Poaceae (15 species), Asteraceae (9 species),
Onagraceae (8 species) and Polygonaceae (7 species).
Through the dendrogram generated in the analysis was possible to verify a cluster of four macrophyte stands located in an oligotrophic region of the reservoir and characterized by the presence of species such as
Nymphoides indica , Nymphea caerulea and Utricularia foliosa . Another cluster grouped 14 macrophyte stands located in mesotrophic or eutrophic regions of the reservoir where were frequently found Salvinia herzogii ,
Lemna aequinoctialis , Pistia stratiotes , Polygonum ferrugineum and Egeria densa . Two macrophyte stands were very different from all the others: one had the lowest species richness and the other had the highest number of endemic species. The last was located within a protected area, the State Park Guarapiranga. With these results it can be concluded that there is spatial heterogeneity of aquatic macrophytes in Guarapiranga reservoir and this distribution is related to the trophic level.
P0141 – ePoster
Distribution, habitat adaptation and conservation of
Anthocleista
species in Nigeria's Niger Delta
Nkang, A 1 , Edwin-Wosu, N 1 , Omara-Achong, T 2
1 University of Calabar, Nigeria;
Nigeria
2 Research Institute,
The Niger Delta is a center of endemism for Africa and is the most extensive and lowland forest/aquatic ecosystem in West Africa. The area is undergoing conversion at a rapid rate. Investigations were carried out on the diversity and abundance of four commonly occurring species of the genus Anthocleista in Nigeria’s Niger
Delta, namely Anthocleista djalonesis A.Chev., A. vogelii
Planch., A. nobilis G.Don and A. liebrechtsiana De Wild.
All the species are found in the wild. Assessment of distribution indicated that the species are generally mesophytic in habitat requirements. Examination of morphological features and geographical distribution patterns indicated that the species are perennial trees with marked preference for tropical climates. Disparities in ecological habitats were found. Anthocleista djalonesis , and A. nobilis show preference for both normal terrestrial
(lowland dry rainforests) and wetland (seasonally flooded) environments. Anthocleista vogelii shows a preference for normal terrestrial habitats while A. liebrechtsiana prefers wetland or semi aquatic habitats.
405

Also, Anthocliesta djalonesis , A. nobilis and A. vogelii are prevalent in lowland secondary rainforests while A. liebrechtsiana is prevalent in fresh water swamp forests.
Anthocliesta leibrechtsiana seems to be a ‘habitat specialist’, showing a narrow niche and limited geographical distribution. The present study revealed declining abundance and distribution of Anthocleista species. The lack of a concise conservation programme in the region threatens these species and other plant genetic resources with extinction. The study highlights the need to promote the development and conservation of these and other economically useful plant genetic resources of the Niger Delta.
P0142 – ePoster
Fine root biomass and its spatial, topographical, and regional variations in tropical rain forests of Brazilian
Amazon
Noguchi, H 1 , de Souza, S 2 , Suwa, R 1
1
Ishizuka, M 1 , Higuchi, N 2
, Kajimoto, T 1 ,
Forestry and Forest Products Research Institute,
Tsukuba, Japan; 2 Instituto Nacional de Pesquisas da
Amazónia, Brazil
Amazonian forest has prominently huge stock of carbon, and has large effect on global carbon dynamics. Fine roots less than 2mm in diameter do not contribute much to the whole carbon stock, and neglected in most biomass studies. However, they are one of the most dynamic parts of trees, which account for 30-50% of the net primary production of forest ecosystems (e.g. Vogt et al., 1996).
CADAF is a joint project between Brazil and Japan, which aims to develop precise and repeatable evaluation technique on a Carbon Dynamics of Amazonian Forest with exhaustive ground data. As a part of this project, we focused on the fine root biomass and its spatial, topographical, and regional variations in tropical rain forests of Brazilian Amazon, which is a basis to clarify the hidden half of the carbon flow of this ecosystem. Fine roots were sampled by steel cylinders in different depth and spatial scales (N = 468 in total) in undisturbed tropical rain forests in S&atilde;o Gabriel da Cachoeira
(0 0 20’ S, 67 0 10’ W), and in Manaus, Brazil (ZF-2 research forest; 2 0 37’ S, 60 0 08’ W). Fine roots were aggregated to shallow top soils, and more than 80 % of their biomass was confined in 20cm depth in both regions. This tendency was more conspicuous at
S&atilde;o Gabriel da Cachoeira forest, which resulted in smaller fine root biomass in this region than in Manaus.
Local variation of the fine root biomass was significantly high within and among 20 x 20m quadrates, but not between longer distances (ca. 300m). Vertical distribution pattern of fine roots also differed among quadrates in both regions. The biomass of fine roots differed among topography, which was higher in lower slope basin with seasonally inundated sandy soil than in plateau with clayish soil.
P0144 – ePoster
Primary pollinators of
Erytrhochiton gymnanthus
Kallunki: hummingbirds
Ocampo, M 1 , Ortega, R, Rodríguez, J
406
1 Universidad Estatal A Distancia, Costa Rica, Central
America;
America; 3
2 Costaricabird.org, Costa Rica, Central
Universidad de Santiago de Compostela,
España
The threat to plants increases as knowledge gaps continue to persist on its ecology; these gaps include those associated with reproduction and interaction with other organisms, which may represent key elements for its preservation in situ or ex situ. This condition is more significant for rare or endemic species, whose vulnerability can be considered synonymous with extinction. In Costa Rica, there is a broad taxonomic knowledge of the flora (about 12,000 taxa); one thousand two hundred taxa are estimated to be endemic. However, there is limited information to determine the actual state of conservation of many endemic species. This research is part of a pioneering study on Erythrochiton gymnanthus Kallunki. The dynamics of growth, phenology, interspecific relationships and genetic variability between and within populations were analyzed for several years. This study identifies 10 species of hummingbirds as primary pollinators of Erythrochiton gymnanthus , at Carara National Park. This shrub is endemic to Costa Rica, belongs to the family Rutaceae and little is known about the populations in the Central and North Pacific of the country . Its condition as sciophilous limits their natural distribution, since determining the relationship with pollinators and dispersers is critical to ensure the stability of their populations . The behavior of hummingbird species varied throughout the observation periods (5:45 to 9:45,
13:00 to 16:45), due their preference for flowers of plants grown in darker areas of the forest, and flowers located at higher altitudes. The interaction of Erythrochiton gymnanthus with pollinators or dispersers had never been documented. This research aims at stimulating the development of knowledge on species considered rare bio-geographically, such as Erythrochiton gymnanthus , in order to provide valuable data to its natural history.
P0145 – ePoster
Habitat conditions and maintenance of
Betula
(birch) forests in the pacific side of eastern Japan
Ogawa, S 1
1 Graduate School of Horticulture, Chiba University,
Tokyo, Japan
Betula spp. are key forest species in deciduous broadleaved forests on the Pacific side of eastern Japan.
However to date, there have only been a few reports on the habitat conditions and maintenance of Betula forests in Japan. We surveyed the habitat condition and maintenance of Betula ( Betula davurica, B. platyphylla ,
B. grossa and B. schmidtii ) forests in the Chichibu
Mountains, eastern Japan. In the survey area, Betula forests of B. davurica , B. platyphylla and B. grossa were distributed only on the stony soil and site on the gentle slope originated from the landslide in the area dominated by Quercus serrata . Those landslide sites were formed few tens years ago. Ages of Betula trees are consisted with landslide event year. The formation of Betula forests depends on the formation of open site due to geomorphic disturbances of landslide. The survey area

often experience geomorphic disturbances of landslide by mechanism and process of landforms of the Sambagawa metamorphic rocks, resulting in the formation of open site by landslide event. Betula forests should maintain their regeneration with frequent geomorphic disturbances in the area. B. schmidtii is different from B.
davurica , B. platyphylla and B. grossa in species characteristics. It is a slow-growing species with a long life span, ca. 500 yrs. It occurs only as dwarf forests on the habitat conditions comprising stony soil and local gentle slopes on the steep ridges in the Chichibu Paleo-Mesozoic area. B. schmidtii trees require such habitat conditions as no soil disturbance, and steep ridges for optimal growth. B.
schmidtii forests maintained in different there less two soil environments: stony soil (type 1) and plentiful soil
(type 2). The forests on type 1 soil consisted of sapling and adult trees, whereas the forests on type 2 soils consisted of only large adult trees. We concluded that B. schmidtii forests were maintained in forests showing predominantly successive regeneration (type 1) but not in forests showing gaps regeneration (type 2).
P0146 – ePoster
Relationship between pollen assemblage and vegetation in Lagos Lagoon, Nigeria
Ogundipe, O 1 , Adekanmbi, O 1
1 University of Lagos, Nigeria
Eight lagoonal communities: Atlas Cove, Five Cowrie
Creek, Maroko, Berger De Motz, Palaver Island, Bayeku,
Majidun Lagoon, and University of Lagos Lagoon were sampled quantitatively with respect to their flora diversity and palynomorphs content. The pollen species encountered range from 10 at Moba, 12 at Five Cowrie
Creek, 28 at Palaver, 37 at Bayeku, 38 at Berger De Motz and 39 at Atlas cove, 52 at University of Lagos Lagoon and 60 at Majidun Lagoon vegetation communities. A total of 96 palynomorphs were recorded in all the sites.
Absolute counts and relative percentages of the vegetation were estimated. The phytosociological record of the eight sampled sites compared favourably with their respective pollen spectra obtained from sediment samples. The dominant plant types are well represented in the pollen spectra in most cases, although some species presented cases of overrepresentation and under representation. The palynofacies are made up of basically pollen grains sourced from local vegetation and only one regional plant. The results of this work show that palynomorphs from Lagos Lagoon sediments truly represent the vegetation and just as it is being used in the
Niger Delta and other places; it can as well be used to characterize parent vegetation communities. Typical swamp pollen grains found in Lagos Lagoon and their botanical affinity are presented as part of this study.
Keywords: flora diversity, palynomorphs, pollen spectra,
Lagos Lagoon.
P0147 – ePoster
Evidence of plant carnivory in
Philcoxia
(Plantaginaceae): underground leaves for nutrient acquisition from nematodes
Pereira, CG 1 , Fritsch, PW 2 , Oliveira, RS 1
1 Plant Biology Dept, Campinas State University –
Unicamp, São Paulo, Brazil; 2 Dept of Botany, California
Academy of Sciences, San Francisco, USA
Plants have evolved a remarkable suite of mechanisms for acquiring growth-limiting nutrients in nutrientimpoverished soils. In the campos rupestres of Central
Brazil, a mosaic of rock outcrops and shallow white sands, soils are highly weathered and impoverished in P and N, and paradoxically, sustain a high diversity of plant species. Very little is known about the diversity of adaptations that allow species to grow and survive in these habitats. Carnivory is considered an effective strategy for plant nutrition acquisition in nutrient-poor and well-lit environments, having evolved independently at least six times in five angiosperm orders. Philcoxia , with three species endemic to the campos rupestres of
Central Brazil, has several unusual morphological features similar to some carnivorous plants, especially peltate leaf blades with conduplicate vernation, stalked capitate glands on the upper leaf surfaces, and leafless scapose inflorescences. An additional and striking feature is the subterranean placement of many of the leaves, which are minute (1.5–3 mm blade diameter) and well imbedded in the white sand in which these plants grow.
The combination of these features, the highly nutrientimpoverished substrate, and the observation of nematode worms sticking to the surface of most leaves in the field prompted the hypothesis that Philcoxia is carnivorous, trapping nematodes and perhaps other soil organisms with the leaf glands and absorbing their nutrients. To test this hypothesis we conducted a greenhouse experiment on one of the species, P. minensis , by hand-feeding nematodes labeled with 15N. Plants were collected from the field and grown in 5-liter pots (one plant per pot) for five months in the greenhouse prior to the experiment.
Ten plants were fed with 15N-labeled nematodes and their leaves harvested 24 and 48 hours after nematode addition. Plants with unlabeled nematodes and those without nematodes were used as controls. Leaves were then oven-dried, cleaned and analyzed for the detection and rate of 15N uptake. Significant amounts of 15N were traceable in nematode-fed leaves in both periods: about
5% of the prey 15N was found in the leaves within 24 hours and 17% after 48 hours. Scanning electron microscope images of P. minensis and P. bahiensis demonstrated the presence of different species of nematode worms at the original sites of both species.
Nematodes were also visible at 60 x magnification with light microscopy on the leaves of the type specimen of the third species, P. goiasensis . Our results support carnivory in Philcoxia , specifically via trapping and nutrient uptake by specialized underground leaves.
Although carnivory is present in two other families of
Lamiales (Byblidaceae, Lentibulariaceae), it has not been previously known in the Plantaginaceae. They also suggest that prey use is critically important as a nutrient source for Philcoxia and uncover a fascinating complement to the diversity of adaptations that allow plants to grow and survive in the species-rich habitats of the campos rupestres .
P0148 – ePoster
Fungal community profile associated to
Epidendrum spp. (Orchidaceae) root system analized by dgge
407

Oliveira, S 1
Pereira, O 3
, Bocayuva, M 1
, Kasuya, M 1
, Veloso, T 1 , Bazzolli, D 2 ,
1 Laboratory of Mycorrhizal Associations, Dept of
Microbiology, BIOAGRO, Federal University of Viçosa
(UFV), Viçosa, Brazil; 2 Laboratory of Genetic and
Molecular Biology of Micro-organisms, Dept of
Microbiology, BIOAGRO, Federal University of Viçosa,
Brazil; 3 Dept Plant Pathology, UFV, Viçosa, Brazil
Orchidaceae is one of the largest families of plant kingdom and includes epiphytic, lithophytic and terrestrial species. A huge diversity of orchids has been observed across the biomes; however, deforestation and the overcollecting have declined this diversity, threatening some species with extinction. In nature, orchids establish an association with mycorrhizal fungi, which is essential for seed germination, vegetative development during the heterotrophic stage of protomeristem and maintenance of individual autotrophic. Although, the Epidendrum is widespread occurring in diferent phytophysiognomies of Atlantic
Forest, these orchids species has shown quite specific in their mycorrhizal associations and are often associated with Epulorhiza fungi. However, different microorganisms, including fungi and symbioticor nonsymbionts bacteria have been observed in orchids.
Therefore, the knowledge of the profile of the microbial community that maintains association with the orchids is important for understanding the ecology and complexity of symbiotic associations. The DGGE (Denaturing gradient gel electrophoresis) was used to evaluate the profile of mycorrhizal fungi associated with roots of orchids of the genus Epidendrum . Fragments of DNA with high intensity in the DGGE gel were sequenced. By
PCR-DGGE it was visualized the profile of Ascomycota and Basidiomycota communities associated in the roots of Epidendrum . It was possible to identify differences between the fungal communities’ profiles from roots of orchids of different localities, Epidendrum spp., and habitat. Using BLAST program, beside Rhizoctonia-like fungi it was also identified Aspergillus genus with 99 % of identity, which has been found as endophytic in other plants. We can conclude that the community of fungi associated with Epidendrum spp. roots is much more diverse than previously demonstrated using the techniques of isolation in pure culture. (Support: AOS,
CNPq,FAPEMIG)
P0150 – ePoster
The Brazilian Atlantic Rainforest with bamboos: a problem for the tree structure?
Padgurschi, MCG 1 , Joly, CA 1
1 Plant Biology Dept, Unicamp, Campinas, Brazil
The Brazilian Atlantic Rainforest is an important
Tropical Forest being classified as a biodiversity hotspot due to high species richness, high degree of endemism and high vulnerability, since there is less than 15% of its original area. The largest remnants of this forest are preserved in São Paulo state, mainly in the slopes of the
Serra do Mar (a range of mountains parallel to the coast that reaches 1,500m above sea level). The biggest conservation unit of the Brazilian Atlantic Forest is the
408
Serra do Mar State Park (PESM, in Portuguese), which is also the largest continuum of remnants of this biome.
Within the PESM, near the border with Rio de Janeiro state, the Atlantic Forest is divided in, at least, 4 physiognomies, classified according to altitude: Restinga
Forest at sea level, Lowland Forest from 50 to 100m,
Submontane from 100 to 500m and Montane Forest from
500 to 1.200, where this study was conducted. The
Brazilian Atlantic Forest is the main center of diversity of a plant's group typical from tropical forests: the bamboos (Poaceae: Bambusoideae). Some studies have showed that when bamboos are present in a high density, forest tree structure changes, with reductions in density, height and diameter of trees. However, these studies were developed in areas that somehow favored the establishment of bamboos (as the semideciduous forest and the open evergreen forest). Our objective was to study an area of Montane Ombrophilous Dense Forest (at
1,100m above sea level) with bamboos to answer the following questions: how is the structure of an Atlantic
Montane forest with bamboos in a well protected and undisturbed area? Is it different from other Atlantic
Montane forests studied in southeastern Brazil? Is there a relationship between bamboos and trees? We carried out our study in an area of Montane Atlantic Forest located within Núcleo Santa Virgínia/PESM (45˚W 04' 05' and
23˚S 19' 32'), using 1 ha plot divided into 100 subplots of
10 x 10 m. Within this plot we marked, plotted in a map and measured (diameter and height) all trees with at least
4.8 cm of diameter at breast height (DBH). For the bamboos, the position of the clumps in the plot and the total area they where occupying within each subplot were recorded. We also counted the number of culms per plot.
We found 1974 trees and 579 clumps/3813 culms of the bamboo Merostachys neesii Ruprecht (Poaceae:
Bambusoideae). Trees DBH ranged from 4.8 to 109 cm, and the average height was 9.1m, (ranging from 5 to 30 m). The presence of bamboos was registered in 94 subplots and we did not find a relationship between the density of culms and the diameter and height of trees.
Comparing with other studies carried out in a Montane
Ombrophilous Forest in São Paulo state (Brazil) we concluded that the presence of bamboo M. neesii in our plot did not altered the structure of the arboreal component of the forest.
P0153 – ePoster
The terrestrial plant communities of Keller Peninsula,
King George Island, Antarctica in the atypical austral summer 2009/2010
1
Pereira, AB 1
CVS 1
, Francelino, MR
, Roesch, LW 1
2 , Moura, TA 2 , Gastal Jr,
Universidade Federal do Pampa, Brazil; 2 Universidade
Federal Rural do Rio de Janeiro, Brazil
This work presents the study of plant communities carried out in the Keller Peninsula, King George Island,
Antarctica, during the atypical austral summer of
2009/2010 and compares the actual results with those from the same area obtained in 2002/2003. Plant communities’ identification was based on their biodiversity, physiognomy, the relationship with bird communities and ground configurations. The area was mapped based on a systematic evaluation and by using an

Astech Promark II® DGPS. By comparing the actual data with the data obtained in 2002/2003 it was possible to observe a large reduction of the area covered by the vegetation. However most of the communities were covered by ice, which avoided a precise determination of the plant community. The 2009/2010 summer presented the lowest temperatures of the last 65 years. The communities were described according to the representative plant species or most abundant biomass and were nominated as: 1. Mosses and flower plants community; 2. Lichens and mosses community; 3.
Leptogium community; 4. Cushion mosses community;
5. Carpet mosses community; 6. Deschampsia and mosses community; 7. Neuropogon community; 8.
Polytrichum community. The results also include distribution maps of the community accompanied by the description of them. This work was supported by the
Brazilian Antarctic Program through CNPq (process no.
574018/2008, FAPERJ (process no. E-26/170.023/2008)
Ministry of science and Technology – MCT, Ministry of
Environment – MMA and CIRM.
P0154 – ePoster
Specialised symbioses and their role in rarity in orchids
, Peakall, R 1 , Barrett, M 4 , Dixon, K 4 , Phillips, R 1,2, 3
1
Hopper, S 4
3
Australian National University, Canberra ACT,
Australia; 2 Kings Park and Botanic Gardens, Perth, WA;
University of Western Australia; 4 Royal Botanic
Gardens, Kew, UK
The role of specialized plant-pollinator and plantmycorrhiza relationships in plant species rarity is poorly understood and has rarely been tested. Here, we test the role of these symbioses in rarity in Drakaea , a genus endemic to south-western Australia, where five of the nine extant species are rare and endangered. Drakaea are pollinated by sexual deception of male thynnid wasps and rely on mycorrhizal fungi for germination and annual growth. Using sequencing of the ITS region and germination experiments, all Drakaea were shown to utilize a single, widespread species of mycorrhizal fungus. Further, in situ baiting demonstrated that germination frequency in both common and rare
Drakaea was similar to that exhibited by co-occurring orchid genera. Baiting for pollinators and DNA barcoding demonstrated that most Drakaea species rely on a single pollinator species. Comparative pollination rate and pollinator abundance data was used to test for general trends between common and rare Drakaea . Fruit set was high in most species but low in a subset of rare species. Baiting at 290 locations across south-western
Australia demonstrated that rare Drakaea tend to have rare pollinators that can limit their expansion into other areas of suitable habitat. At regional scales, some pollinators of rare species were more widespread than the orchid demonstrating that pollinators are not limiting their distribution. While there are trends within the genus, considering the ecology of each species demonstrates that in Drakaea the causes of rarity are idiosyncratic. Species rarity appears to result from different combinations of the abundance of pollinators, habitat specialisation and the evolutionary history of each
Drakaea species.
P0155 – ePoster
Influence of heavy metal toxicity on defense gene activation in
Arabidopsis thaliana
affected by necrotrophic fungal attack
Cabot, C 2 , Gallego, B
Poschenrieder, C 1
1 , Martos, S 1 , Barcelo, J 1 ,
2
1 Universidad Autónoma de Barcelona, Spain;
Universidad de las Islas Baleares, Spain
Abiotic and biotic stress share common signaling pathways. High, potentially toxic concentrations of heavy metals have been described as elicitors of plant defense reactions similar to those induced by microbial infections. However, very few studies have addressed the interactions between high metal concentrations and pathogen infections on stress signaling in plants . More detailed investigations into the mechanisms of such interactions will be useful not only for a better understanding of the fundamental processes governing specificity and cross-talk in stress signaling pathways, but also may help in the future development of both better stress adapted plants and more environmentalfriendly and efficient phytosanitary products. Here we report first results on the influence of Zn and Cd, alone and in combination with inoculation with necrotrophic fungi, on the expression of genes related to jasmonate and salycilate signaling in Arabidopsis thaliana . Plants were pre-cultured in hydroponics using control nutrient solution or solution spiked with either 10 µM Zn or 10
µM Cd. Part of the plants receiving excess Zn were inoculated with the necrotropic fungus Alternaria brassicicola , while part of the Cd-stressed plants were infected with Botrytis cinerea . RT-PCR were performed to reveal the level of expression of PR1 and PDF1.2 as markers for the salycilate and jasmonate–dependent signaling pathways, respectively. Cadmium and zinc alone had no effect on the expression of these stress related genes. Infection with necrotrophic fungi typically enhanced expression of the jasmonate signaling related gene PDF1.2, while the salycilate- related PR1 was not enhanced. Alternaria brassicicola was more effective than Botrytis cynerea in inducing PDF1.2. Infection with
Alternaria in plants exposed to excess Zn exhibited an even stronger induction of PDF1.2( more than 250 times increase). In plants simultaneously exposed to excess Zn and Alternaria infection also PR1 expression was enhanced (8 times). According to these first results excess Zn supply can enhance the resistance response of
Arabidopsis thaliana to Alternaria brassiciola . In contrast, our results do not support the view that Cd acts as elicitor of defense responses in this system.
Acknowledgements: Supported by the Spanish Ministry of Science and Innovation projects
P0156 – ePoster
Liana success, ecophysiology of two
Bauhinia
species in Hong Kong
Ramsden, L 1 , Luk, M 1
409

1 University of HongKong SPACE, School of Biological
Sciences, China
The woody climbing plants or lianas are an important component of vegetation in Hong Kong and can constitute a considerable quantity of the biomass in their natural habitats. In this study the physiological factors which underlie the success of two contrasting liana species within the same genus, Bauhinia glauca and
Bauhinia championii have been investigated. Previous research had shown that although these two lianas have an overlapping distribution in HK their reproductive biology shows differences that are likely to represent adaptations to different environments (Lau et al. 2009).
In Hong Kong both lianas are an abundant part of hillside vegetation and appear to occur in similar locations, yet closer examination revealed that only B. championii can persist in more densely vegetated natural forest while B. glauca only occurs in exposed sites, where however, B. championii can also be found. Historical records indicate that B. championii was considered to be the rarer of the two species but its abundance is now increased.
Examination of photosynthesis in the two species failed to find a clear difference that could support different adaptations, with similar light responses and maximal efficiency observed. Study of water relations revealed a higher resistance to water loss in B. championii leaves with several aspects of morphology contributing to improved water control compared to B. glauca which was vulnerable to water stress. But how can this be reconciled with the apparent preference of B. glauca for more exposed sites?
P0157 – ePoster
Forest cover change detection using remote sensing and geographical information system
Rane, Gauri
Kumar, A 1
1 , Joshi, B 1 , Mahendran, V 1 , Mahajan, C 1 ,
1 JalaSRI, Watershed Surveillance and Research Institute,
Maharashtra, India
Jalgaon is located at 20º-27ºN, 74º-76º E in the North-
West region of Maharashtra state, India. Total forest area of the district is 1185 sq. km comprising 10.07% of the total geographic area and divided into scrub forest on the southern side of river Tapi and dry deciduous forest on the northern side of river Tapi. Dry deciduous forest is having a continuous forest patch while the scrub forest on north of river Tapi is fragmented and divided into smaller patches. The forest area has been found to be reduced from 2000 onwards. The temperature of the
Jalgaon district is however, increased and rainfall is reduced which indicates that deforestation can be one of the associated causes for it. Forest can be considered as one of the ecosystem having biological diversity as it supports the life. Forest plays an important role for maintenance and enhancement of environmental quality, social security of livelihood and economic development.
Forest land is decreasing continuously due to encroachment by the tribal for the agricultural practices.
Land use and Land cover mapping is really important to detect the neighborhood, which in turn help us to study ecosystem functioning, biophysical and human variables.
The changes of Land use and Land cover pattern over a
410 time period control the pressure on land. Land cover data presents the physical coverage of the earth’s surface while Land use data presents its socio-economic functionality. For the present study IRS P6, LISS-III data of November 2007 and January 1998 is used to detect land cover changes and forest status using Remote
Sensing, Geographical Information System and GPS tools. Study area makes up most of landuse and landcover classes like agriculture, forest, barren land and water bodies. Though, Jalgaon is developing rapidly but still agriculture retains the major land use class followed by barren land, forest covers, settlement and water body respectively. The study reveals a positive change in the overall forest cover with 1.02%. The forest area in 1998 was 1153.53 sq. km and it is 1185 sq. km in the year
2007 with a significant growth of 2.73 %. This positive change can be associated with new plantation as well awareness policies to protect the forest by centre and state governments. Dense forest observed the increase of
4.34% with an area of 415 sq. km and 397 sq. km respectively in 2007 and 1998. Open forest in other hand decreased 6 sq. km during the same period. A significant loss has also been mapped in scrub forest area with a
31.68% during the same period.
P0158 – ePoster
Medicinal plant biodiversity of Chennimalai and its
1 conservation
Ravi Raja,S 1 , Asir Selin,K 1
Scott Christian College, Nagercoil, India
Chennimalai is popular for its rich medicinal and aromatic plant diversity. The varying soil, topography and occurrence of different climatic and microclimatic zones of the region provide very congenital conditions to grow a number of therapeutically important medicinal plant species. In ancient times many Siddhar and medicinal practitioners were utilizing many plants of this hills for medicinal uses. In recent years due to increasing awareness about herbal products there is tremendous pressure upon medicinal plants. In this region, no serious attempts are made so for on cultivation of these important plants, especially medicinal and aromatic plants. The unscientific, over and irregular exploitation of medicinal plants from their natural habitats has resulted in very fast depletion as well as extinction of some important species like, Eltyraria acaulis , Evolvulus alsinoides , Ionidium suffruticosum , Randia dumetorum ,
Canthium parviflorum etc. So urgent need for conservation of many such species by adopting scientific methods are required.
P0159 – ePoster
1
Microsatellite markers reveal that a? small translocated population of
Dianthus superbus
L. has not lost genetic diversity
Reier, Ü 1 , Rammul, K 1 , Talve, T 1 , Oja, T 1
Dept of Botany, Institute of Ecology and Earth Sciences,
University of Tartu, Estonia

Low level of genetic diversity is expected in small populations; including those that have undergone bottleneck or substantial reductions in abundance.
Dianthus superbus L. (Caryophyllaceae) is a perennial outcrossing meadow plant. Its formerly large continuous populations are nowadays small and isolated, mostly due to anthropogenic activities. We studied genetic diversity of a translocated population and compared it with three natural populations. The translocated population is a fragment of the population now appering to grow within the limits of the town (Tartu, Estonia). We used DNA microsatellite markers designed for genus Dianthus
(Smulders et al. 2000, Smulders et al. 2003). 59 alleles were detected (10 loci) of which four were unique for the small translocated population. The number of unique alleles did not differ significantly from the natural populations as well as the average allele number per loci
(3,6 for transplanted population and 3,6 for natural populations). The results showed quite high genetic diversity in the small translocated population. Our results suggest that the small translocated population of
Dianthus superbus has not critically lost its genetic diversity and does not need immediate augmentation with plants from bigger populations. The translocated population sets viable seeds and has given more than 180 seedlings of which two have reached generative stadium in five years after translocation. The translocated population had the highest inbreeding coefficient (0,499) in comparison with the three natural populations (0,194;
0,228; 0,269). This is inevitable because of the small number of parent plants and geographical isolation from the nearest population. The distance of about one kilometer is crucial for insect-pollinated species with no means for long-distance seed dispersal. This can lead to inbreeding depression in longer perspective.
P0161 – ePoster
Triplaris americana
L. (Polygonaceae) sex ratio at the
Brazilian Pantanal
Reis, T 1
1 Universidade Estadual de Campinas, Brazil
Dioecious plant populations can have their sex ratio skewed towards one gender. These deviations may have an environmental basis, such as differential mortality of staminate (male) and pistillate (female) plants. Likewise, different ages of sexual maturation between sexes may increase the incidence of one sex over another. Both factors may vary with habitat, so that same species in distinct habitats may have different sex ratios. Moreover, investment in reproduction often differs between sex and in most cases females allocate more energy to reproduction. According to the reproduction cost hypothesis, this can compromise other female life history aspects, such as growth and survival. In this case, we would find smaller sizes for females and male biased sex ratio. Triplaris americana is a dioecious tropical tree that occupies habitats susceptible to flooding and high luminosity. In the Miranda Sub-region of Pantanal this species can be found both in the riparian vegetation along the river Miranda and in natural forest fragments. These two habitats differ in their resource availability and the riparian forest would be a more favourable environment for T. americana considering the high demand for flooding in the life cycle of this species. For that matter, it would be expected a less skewed sex ratio for males in the riparian forest following the existing trend of staminate flowers predominating in stressful habitats and pistillate flowers in favorable ones. Thus, the aims of this study were: (1) compare the sex ratios of T. americana among habitats, and (2) investigate possible relationships between size and sex of individuals in both habitats. No sex ratio deviations were found in any of the habitats.
This is consistent with the lack of evidence for both differential mortality and differential sex maturation between sexes since the distributions of plant basal area were similar between male and female plants. Thus, these factors that would deviate sex ratio were canceled for these T. americana populations, which could be indicating possible compensation mechanisms for the costs of female reproduction (e.g. increased photosynthetic rate) although this could not be evaluated in this study. However, another ecological factor that could lead to biased sex ratio is the difference in vegetative reproduction between sexes. More investment in vegetative reproduction for males and sexual reproduction for females are usually found. This also seems to occur for T. americana since the number of stems per individual were higher in males. This investment could mean increases in male survival and possible sex ratio deviations, although these have not been found. Finally, if we consider the existence of mechanisms by which female plants may be offsetting the cost of reproduction, smaller sizes for females would not be expected. Indeed, the riparian female plants were even higher than male ones, which may be related to this species anemochory. Height increases for female plants would be an advantage in seed dispersion. Even with this apparent strategy of vertical expansion for females and horizontal for males (through vegetative reproduction), there were no differences between sexes with respect to individuals allometry.
P0162 – ePoster
Spatial pattern of the sub-canopy tree
Bathysa australis
(A. St.-Hil.) & Hook. in an Atlantic tropical rain forest of SE Brazil
Reis, TS 1 , Costa, RC 1 , Santos, FAM 1
1 Unversidade Estadual de Campinas, Brazil
The spatial pattern of a plant population is related to the spatial and mortality patterns along its life cycle. The spatial distribution of small individuals is most influenced by the dispersal patterns, but also by the distribution of safe sites. In the following stages, it is more likely that environmental constraints are more important in determining distribution patterns since environment induced mortality may already be occurring.
In this case, the tendency would be the formation of aggregates at favorable sites. However, high competition in these sites would favor density-dependent mortality so that survival would be associated with decreasing aggregation. Assuming resources have a clumped distribution, it is expected that aggregation also occur for juvenile and adult stages although this would be less intense considering the presence of density-dependent mortality. Bathysa australis (Rubiaceae) is a sub-canopy tree whose light-demanding germination and restricted
411

dispersal distances suggest that local scale plays an important role in both the spatial distribution and population dynamics. This species occurs at Atlantic
Tropical Rain Forest of Serra do Mar State Park (23° 31' to 23° 34' S and 45° 02' to 45° 05' W), where this study was performed. Sampling consisted of three 1ha plots in the upper forest (1,000m a.s.l.) and three 1ha plots in lower forest (100-200m a.s.l.). All individuals were mapped, measured at diameter at ground level (DGL) and classified into three size categories: small
(DGL<15mm), intermediate (15
≥
DGL
≤
50mm) and large
(DGL>50mm). We analyzed the spatial patterns with
Ripley's K univariate function and tested for associations between size classes with the Ripley's K bivariate function. We sampled 636 individuals of B. australis in the upper forest and 920 in the lower forest. Individuals of the smaller size class were aggregated in all plots and positively associated with the largest individual’s class in most scales, except for one plot. This pattern is consistent with the limited seed dispersal of this species and suggests the importance of seed dispersal process in the spatial distribution of smaller size individuals. We also found a trend of decreasing aggregation intensity with ontogeny. The smallest size class had always the highest level of aggregation, which decreased in subsequent classes in most of the plots, getting random in one lower plot. Association tests still showed positive associations between small and intermediate individuals and between intermediate and large ones for most of the plots.
However, only in two plots the association between large individuals and intermediate ones has become weaker in relation to the association between large and small ones, contradicting the expectation of a decrease in aggregation intensity due to intraspecific competition. For Bathysa australis the negative effects of intraspecific competition may not be so intense as to overcome the positive effects of the occupation of favorable sites. Finally, this distribution in relation to favorable sites seems to be more limiting for small and intermediate individuals since the association between those size classes was the most intense. All results hold for lower and upper forest plots in spite of their altitude differences.
P0163 – ePoster
Triplaris americana
L. (Polygonaceae) sex ratio at the
Brazilian Pantanal
Reis, TS 1
1 Unversidade Estadual de Campinas, Brazil
Dioecious plant populations can have their sex ratio skewed towards one gender. These deviations may have an environmental basis, such as differential mortality of staminate (male) and pistillate (female) plants. Likewise, different ages of sexual maturation between sexes may increase the incidence of one sex over another. Both factors may vary with habitat, so that same species in distinct habitats may have different sex ratios. Moreover, investment in reproduction often differs between sex and in most cases females allocate more energy to reproduction. According to the reproduction cost hypothesis, this can compromise other female life history aspects, such as growth and survival. In this case, we would find smaller sizes for females and male biased sex ratio. Triplaris americana is a dioecious tropical tree that
412 occupies habitats susceptible to flooding and high luminosity. In the Miranda Sub-region of Pantanal this species can be found both in the riparian vegetation along the river Miranda and in natural forest fragments. These two habitats differ in their resource availability and the riparian forest would be a more favourable environment for T. americana considering the high demand for flooding in the life cycle of this species. For that matter, it would be expected a less skewed sex ratio for males in the riparian forest following the existing trend of staminate flowers predominating in stressful habitats and pistillate flowers in favorable ones. Thus, the aims of this study were: (1) compare the sex ratios of T. americana among habitats, and (2) investigate possible relationships between size and sex of individuals in both habitats. No sex ratio deviations were found in any of the habitats.
This is consistent with the lack of evidence for both differential mortality and differential sex maturation between sexes since the distributions of plant basal area were similar between male and female plants. Thus, these factors that would deviate sex ratio were canceled for these T. americana populations, which could be indicating possible compensation mechanisms for the costs of female reproduction (e.g. increased photosynthetic rate) although this could not be evaluated in this study. However, another ecological factor that could lead to biased sex ratio is the difference in vegetative reproduction between sexes. More investment in vegetative reproduction for males and sexual reproduction for females are usually found. This also seems to occur for T. americana since the number of stems per individual were higher in males. This investment could mean increases in male survival and possible sex ratio deviations, although these have not been found. Finally, if we consider the existence of mechanisms by which female plants may be offsetting the cost of reproduction, smaller sizes for females would not be expected. Indeed, the riparian female plants were even higher than male ones, which may be related to this species anemochory. Height increases for female plants would be an advantage in seed dispersion. Even with this apparent strategy of vertical expansion for females and horizontal for males (through vegetative reproduction), there were no differences between sexes with respect to individuals allometry.
P0164 – ePoster
Victorian orchid conservation project
1
Reiter, N 1
Pollard, G 1
, Cross, R 2
, Argall, M
, Wright, M 2
1
, Lawrie, A 3
, Bedggood, W 1
, Pitts, D
, Burns, A 1
1 ,
Dept of Sustainability and Environment, Victoria,
Australia;
Australia; 3
2 Royal Botanic Gardens, Melbourne,
RMIT University, Melbourne, Australia
The Victorian Orchid Conservation Project aims to implement a range of actions from the National Recovery
Plans and Flora and Fauna Action Statements of 80 species of terrestrial orchids from across the state of
Victoria. Using ex situ symbiotic germination and reintroduction, this collaborative project lead by the
Victorian Department of Sustainability and Environment also involves the Royal Botanic Gardens Melbourne, the
Australasian Native Orchid Society, the University of
Melbourne, RMIT University and many community

groups. Summaries of four federally threatened orchids from southwest Victoria that have had substantial recovery work conducted on them over the past decade are presented. The work includes surveys, long term monitoring, propagation, habitat modification, reintroductions and mycorrhizal phylogenetic work. The species highlighted here are Caladenia hastata
(Melblom’s Spider-Orchid), Caladenia calcicola
(Limestone Spider-Orchid), Caladenia xanthochila
(Yellow Lip Spider-Orchid) and Thelymitra epipa ctoides
(Metallic Sun-Orchid). Melblom’s Spider-Orchid has been a pioneering plant in much of this conservation research giving valuable insight into ‘how to conduct orchid conservation work’. The Limestone Spider Orchid and Yellow Lip Spider-Orchid have both been monitored over a ten year period and have had successful trial reintroductions conducted in 2007; the information being gathered on these two species will aid future reintroductions of Caladenia throughout Victoria. The
Metallic Sun-Orchid research is investigating the edaphic conditions in which its populations grow, the diversity of its associated mycorrhizal populations both within and between populations, and conditions for optimal ex situ nursery growth.
P0166 – ePoster
Effect of annual plants community and disturbance on ant diversity in a semiarid zone in Central Mexico
Rivas Arancibia, SP 1 , Carrillo Ruiz, H 1
1 Universidad Autónoma de Puebla, Mexico
Ants have a wide geographic distribution, and it is know that local communities are affected by several factors such as temperature, humidity, sunlight incidence and interactions with other organisms. Due to its high sensitivity to environmental changes and the wide range of niches they occupy in the ecosystem, these insects have been proposed as anthropogenic disturbance bioindicators. In Zapotitlan Valley a semiarid region in
Central Mexico, the ants interact with plant communities and affect their distribution, abundance and species composition, by their granivorous habits. This relationship is reciprocal, the diversity and abundance of seeds also affects the diversity of ant colonies. The short periods of seed production of annual plants during the rainy season, could be important in providing insect biomass of the ecosystem. It has been reported that annual plants are colonizing disturbed and early successional habitats, although in the Zapotitlan valley this is not always true and is not yet clear their ecological role. So in this study we worked on the hypothesis that the annual plant community could reduce the negative effects of disturbance on ant communities in those places where they are present, but not for sites with few species of annual plants, these last show a decrease in ant diversity and increased abundance of only a few species.
In the study area with previous information we selected six sites in the Valley of Zapotitlan organized as a 3 2 factorial design. Two groups were formed: three sites with low and three high human disturbances. Each group with three levels of annual diversity (high, low and no annuals). On two parallel transects at each site, placed 20 pitfall traps baited with tuna. The traps were checked twice daily (morning and evening) for five days. We identified 17 species of ants, of which 11 are new records for the Zapotitlan Valley. Preliminary results have shown that the diversity and abundance were higher at sites with low disturbance. The sites with high annuals diversity and with high and low disturbance, showed significant differences only in four ant species that could be considered as anthropogenic disturbance bio-indicators.
The sites with few annual species or without them (with high disturbance conditions) not showed difference on ant diversity or abundance. The methodology does not consider species with activity on the ground surface or above one meter in height which explains the large number of new ant species records. It is necessary to evaluate the combined factors effect to understand and suggest a better explanation for the proposed hypothesis.
P0167 – ePoster
Daily and seasonal water use dynamics of
Calophyllum brasiliense
Cambess. (Calophyllaceae) in a seasonally–flooded restinga forest in southeast
Brazil
1
Saenz, G 1 , Joly, C 1
Unicamp, Brazil
, Oliveira, R 1
Soil water saturation may represent a limitation to plant water acquisition because low diffusion of gases in flooded soils causes anoxic conditions in the rhizosphere.
However, flood-tolerant species have adaptations that may allow maintenance of stomatal conductance and growth during flooding periods. In order to evaluate how flooding affects the water use of Calophyllum brasiliense , we monitored the daily and seasonal dynamics of sapflow of trees in a tropical restinga forest in São Paulo state, Brazil. Restinga forests occur over coastal sandplains and are subjected to seasonal flooding due to the rise of the water table level. We used the heat ratio method (HRM) to make continuous measurement of sap flow. We also monitored continuously the table water level and VPD (vapor pressure deficit) close to the tree crowns. We found evidence of strong stomatal limitation of maximum sapflow rates during periods of both high and low water table levels. On a daily basis, sapflow reached maximum values around 10:30-11 am and decreased during midday, when VPD values exceeded 1 kPa. This indicates an upper limit to transpiration, despite the level of the water table. On a larger time scale, sapflow was positively and highly correlated with
VPD, while the water table level had a minor influence in the water use dynamics of the trees. The results suggest that transpiration of C. brasiliense is not constrained by flooding but rather by high atmospheric demand and limitations imposed by the hydraulic system of the trees.
P0168 – ePoster
Coexistence mechanism of tree species in a natural riparian forest throughout the reproductive traits, on
1 upper stream in central Japan
Sakio, H 1
Niigata University, Japan
413

Reproductive traits and responses to the environment differ widely among tree species. To understand tree coexistence, it is important to investigate the entire life history and demography of coexisting tree species in a community. Water and light are important factors for the growth of trees in the cool-temperate zone. Gap formation accelerates the growth and flowering of trees, and while trees require moderate humidity in soil, flooding may retard their growth. Many tree species
( Fraxinus platypoda , Pterocarya rhoifolia and
Cercidiphyllum japonicum , Acer carpinifolium etc.) coexist on riparian forests in the Chichibu Mountains, central Japan. We compare the reproductive characteristics among tree species. F. platypoda is a dominant species of canopy trees. F. platypoda have large-sized seeds with clear irregular fruiting behaviors with many saplings on abandoned channel and floodplains, and can invade not only large disturbance sites but also small disturbance sites. F. platypoda seedlings had relatively high tolerance for shade and flooding treatments among tree species. P. rhoifolia is a subdominant canopy species that have patch structure containing even-aged trees on the deposits of large–scale landslides. P. rhoifolia have large-sized seeds with irregular fruiting behaviors. Seedling is distributed gravel site under large gaps. In forest gaps, the growth of seedlings of P. rhoifolia was faster than that of F. platypoda.
C. japonicum is the other subdominant canopy species. C. japonicum produce a lot of smallsized seeds every year, and have many sprouts around main stem. This tree had few saplings, and invaded large disturbance sites together with P. rhoifolia . The establishment site of C. japonicum was restricted to fine mineral soil and fallen logs. Sprouts of this tree play an important role in the maintenance of the canopy in the processes of regeneration. A. carpinifolium is dominant lower layer species. This tree had clear irregular fruiting behaviors with few seedlings on forest floor. A. carpinifolium have relative high shade tolerance, and have many sprouts for the maintenance of individual.
The causes of coexistence of the riparian canopy tree species are both niche (niche partitioning and tradeoff)- and chance (disturbance and competition) -determined in variable proportions. There were tradeoffs of reproductive characteristics between the tree species. The tree diversity was maintained in riparian areas through some niche partitioning and different reproductive strategy. Disturbance regime of riparian forest was various in type, frequency and size. On the other hand, chance played a major role in regeneration of P. rhoifolia and C. japonicum through the unpredictable disturbance in large size. In conclusion, these riparian tree species were well-adapted to various disturbances throughout their life-history stages.
P0169 – ePoster
A brief on the floral diversity, threats and conservation activities in the United Arab Emirates
Sakkir, S 1
S 1
, Kabshawi, M 1 , Al Mehairby, M
1 Environment Agency, Abu Dhabi, UAE
1 , Al Dhaheri,
A brief discussion is presented on the current floral diversity of UAE including threat analysis and
414 conservation steps underway. United Arab Emirates, consisting of 7 emirates, with a total area of 83,600 km2 has been an abode of rich biodiversity despite the prevalent hyper-arid conditions.UAE is home to 731 species of vascular plant species (i.e. 386 genera, 83 families) and the 4th largest in the highest number of angiosperms in the Arabian Peninsula. The flora constitutes therophytes (50%), chamaephytes (33.24%), hemicryptophytes (8%), phanerophytes (4.2%), geophytes (3.4%), lianas (0.8%) and parasites (0.4%). A good percentage (18%) of the species is found to have medicinal properties and has been traditionally used in medical practices. The diverse habitats, UAE is blessed with, such as coastal habitats, sabkhas, mountains & wadis, deserts, plains, oases, urban habitat, and offshore islands, each with its own phyto-geography and plant communities account for this diversity. Of these, mountains and wadis (33%) and gravel plains (26%) are found to support distinctly high floristic diversity. And the mangroves on the coastal habitats also merit a special mention. However, given the fragility of arid/semi-arid regions, threats, both man-made and natural pose serious challenges to natural vegetation, plant biodiversity and sustainability of natural environment. Applying IUCN categories, preliminary data reveals 1.5% of flora belongs to threatened category, even when there is no critically endangered species facing extinction. 4.8% of floras are vulnerable to potential extinction. While 77.7% belong to low risk category, data are not available for
2.2% of species. Threat analysis, based on the data available to Environment Agency, Abu Dhabi (EAD) suggest, habitat degradation, desertification, overgrazing, climate change and introduction of exotic species as prominent reasons which play significant role in the deterioration of plant diversity. Through annual surveys during the decade 2001–2010, EAD has covered 60% areas of Abu Dhabi Emirates. As a result, EAD maintains a herbarium of over 3000 specimens and plant nurseries where seeds of native species are collected and propagated. Rich documentation on species-rich areas is also available for research. Under the state sponsored conservation activities (of EAD), both short term and long term strategies are underway. Under short term approaches, specific priority areas are identified and strategies specific to each priority area are being executed. Long term strategies however, focus on generating the right awareness among students and public (including key decision makers, scientists) on environment protection, an enhanced understanding of the biological diversity and uniqueness of the region.
Standard conservation steps such as establishment of botanical gardens, seed bank, and restoration of habitats are already underway. Besides, UAE is signatory to international conventions such as Ramsar Convention on
Wetlands, CITES and CBD.
P0172 – ePoster
Stocks and flows of coarse woody debris in a hill dipterocarp forest in peninsular Malaysia
Sato, T 1
Ripin, A 3
, Niiyam, K 2 , Yagihashi, T 2 , Kassim, ARB 3 ,
1 Forestry & Forest Products Research Institute,
Tsukuba, Japan; 2 Tohoku Research Center, Forestry &

3
Forest Products Research Institute, Morioka, Japan;
Forest Research Institute Malaysia, Selangor, Malaysia
Coarse woody debris (CWD) is a major pathway of carbon flux in forest ecosystems. Because CWD is a large and labile pool of carbon with a significant impact on the net carbon budget of forest ecosystems, surveys of live biomass alone are insufficient to determine carbon budgets. In this study, we investigated the stocks of
CWD in a hill dipterocarp forest in peninsular Malaysia.
The study was conducted in an old-growth hill dipterocarp forest in the Semangkok Forest Reserve,
Selangor, Malaysia. The study plot is dominated by
Shorea curtisii . To estimate the mass of CWD, we employed a sample plot inventory method using thirtyfive 20-m×20-m quadrats. In each quadrat, all CWD with an end diameter
≥
10 cm was measured in September
2009 and September 2010. We also classified CWD type as snaps, standing dead, uprooted logs, or fallen logs.
The total necromass of CWD averaged 81.43 ± 100.38
(SD) Mg ha –1 (range, 0.10–379.73 Mg ha –1 ). This average value is greater than that estimated in a lowland dipterocarp forest at Pasoh (49 Mg ha –1 ). The CWD mass accounted for about 15% of AGB. This percentage is higher than that in other tropical forests. During the census period (1992–2007), the mean input of CWD from tree mortality was 103.4 Mg ha –1 over 15 years
(0.10–379.73 Mg ha –1 ). In the plot, fallen logs were the major form of CWD, accounting for 40%. About 35% of
CWD was uprooted and 18% was snaps. The stock and input of CWD differed markedly according to topographic position in the plot. The headmost wall slope position had a higher CWD mass than that in other slope positions. On the other hand, CWD mass was lower on the lower slopes and bottomland, which were densely covered with palm ( Eugeissona tristis ) and bamboo
( Gigantochloa scortechinii ). In the Semangkok Forest
Reserve, many Shorea curtisii trees are distributed around ridges, and are dominant within the top canopy layer (e.g., 40–50 m in height). This specific stand structure is one of the reasons that plots in hill dipterocarp forest ecosystems tend to have a high CWD stock.
P0174 – ePoster
Mexican wild cactus pear conservation proposals
Scheinvar, L 1 , Gallegos, C 2 , Olalde, G 1 , Sule, D 1
1 Mexican National Autonomuos University; 2 Centro
Regional Universitario Centro Norte, Universidad
Autonoma Chapingo, Mexico
In the Cactological Laboratory of the Botanical Garden of the Biological Institute of the Mexican National
Autonomous University, we are developing a systematic, phytogeographic and ecological project about the
Mexican wild cactus pears species (genera: Opuntia and
Nopalea ). We studied the effects of climatic changes over the species that inhabit in the central region of
Mexico. Cytogenetical studies of the wild cactus pears of
Nayarit and Colima are developed. Initially we revised the cactus pear deposited in 42 Mexican herbaria, taking digital photos of the sheets and revising the identification of each one. Intensive excursions to the field where realized, especially in states and areas poorly collected and poorly represented in the herbaria. In the field plants where georefered, were taken digital photos of their habitat and habit, specially of the stem, cortex, canopy, flowers and fruits. Four cladodes of each studied plant were collected; others were observed and the dates anoted. Their dates where included in a data wich contains 5800 registers that can be consulted in the web page of our Biological Institute. For each species we included a detailed study of microstructures made with the scanning electronic microscope of areoles, thorns, glochids, pollen grains, epiderm and seeds. Our team was composed of 16 students of biology that elaborated thesis and social service in our laboratory, dedicated to the study of the wild cactus pears in the states of Baja
California, Chiapas, Chihuahua, Colima, Guanajuato,
Hidalgo, Nayarit, Oaxaca, Puebla, Querétaro, San Luis
Potosí, Sinaloa, Sonora, Tamaulipas, Tlaxcala and
Zacatecas. We elaborated maps of the geographical distribution of each species and maps of models of potential distribution. This maps where overlapped to the maps of protected natural areas and types of vegetation.
We made also maps for each state with the same information. We elaborated maps with hot spots where cactus pear inhabit in Mexico and predictive maps of potential distribution with the impact of the climatic changes in the future 20 and 50 years in the central states of Mexico. We have in Mexico 174 protected natural areas (CONANP 2002) but only 47 contain wild cactus pear, some are in their bordes. We identified 93 wild species of Opuntia , with 62 endemic and nine microendemic, and 8 wild species of Nopalea , with 6 endemic. Mexico is the center of origin of Nopalea and this genera has only 3 species that extend their distribution to Guatemala and Honduras. It is also an important center of diversification of Opuntia . To conserve the native prickly pear germoplasm the protected natural areas are an important estrategy in-situ and the collections of the Botanical Gardens ex situ. In our Botanical Garden, the 'National Collection of
Mexican Wild Cactus Pear' actually with 8 areas, has 397 plants of 64 species of Opuntia and 3 of Nopalea . We thank the director of our Biological Institute and
Botanical Garden, CONABIO and SINAREFI for the support to this project. With the information obtained we are actually elaborating an Atlas of the wild Mexican cactus pear.
P0176 – ePoster
Genetic variation in photosynthetic response of wheat to elevated CO
2
in Australian grains free air carbon dioxide enrichment (AGFACE)
Seneweera, S 1
Fitzgerald, G 3
, Thilakarathne, L
, Tausz, M 4
1 , Posch, S
, Borgeault, M 5
1 , Norton, R 2 ,
1 Dept of Agriculture and Food Systems, University of
Melbourne, VIC, Australia; 2 International Plant
Nutrition Institute, VIC, Australia;
Primary Industries, VIC, Australia; 4
3 Victorian Dept of
Dept of Forest and
5
Ecosystem Science, University of Melbourne, Australia;
CSIRO Climate Adaptation Flagship, CSIRO
Sustainable Ecosystems,Qld, Australia
The present atmospheric CO
2
concentration is rising and is predicted to double by the end of the 21st century which is likely to have a profound effect on plant growth
415

and yield, especially for C
3
plants. A large genetic variability in growth response to elevated CO
2
(eCO
2
) has been documented among different species and there is very limited understanding of the underlying mechanism. In this study, wheat ( Triticum aestivum L.) was used as a model plant to investigate the physiological basis of the intraspecific variation in plant response to elevated CO
2
. Seven wheat (Triticum, aestivum L.) cultivars, Janz, Yitpi (high tillering),
Silverstar, H45 (synchronous tillering), Drysdale, Hartog, and Zebu (medium tillering) were grown in the
Australian Grains Free Air Carbon dioxide Enrichment
(AGFACE) facility. The effects of rising carbon dioxide
(ambient aCO
2
~380 ppm, elevated eCO
2
~550 ppm) on photosynthesis (A) and its related processes were investigated. Gas exchange measurements of flag leaf blades were carried out at pre and post anthesis stages. A versus intercellular CO
2
concentration (Ci) was used to estimate apparent maximum carboxylation capacity
(Vc.max) and photosynthetic electron transport rate
(Jmax ). A variation in A when measured at growth CO concentration (either aCO
2
or eCO
2
2
) was observed among the cultivars tested where A varied from 20-23 and 20- 29 µmol CO
2
m-2 s-1 at aCO
2
and eCO
2 respectively. The highest pre-anthesis A responsiveness
(eCO
2
/aCO
2
) was 45% for Janz while Yitpi showed least stimulation. In contrast, post anthesis A was significantly decreased for all cultivars when compared to post anthesis measurement where Drysdale and Zebu showed the least reduction. Mechanistic analysis of gas exchange data showed large variation in apparent maximum carboxylation capacity of Rubisco (Vcmax) and photosynthetic electron transport rate (Jmax). A reduction in Vc.max at eCO
2
was prominent at the post anthesis stage suggesting acclimation of A to eCO2.
There were genetic differences in A acclimation to elevated CO largest acclimation (Vc.max at eCO aCO
2
2
among the cultivars tested. H45 showed the
2
versus Vc.max at
) compared to Zebu, which showed the least acclimation to elevated CO
2
. These data suggest that the magnitude of growth stimulation to eCO
2
is closely linked with the capacity of the plant to acclimate photosynthesis to eCO
2
. Acknowledgement Funding from the Australian Commonwealth Department of
Agriculture, Fisheries and Forestry (DAFF) and the
Grains Research and Development Corporation (GRDC) is gratefully acknowledged.
P0177 – ePoster
Species richness in Thadagai Hills of Western Ghats,
Tamil Nadu, India
Sethuraman, P 1
1 Nirmala College For Women, Coimbatore, Tamil Nadu,
India
Western Ghats are themselves renowned as one of the world’s great biodiversity hot spots. This great chain of hills effectively operates as a string of islands, each with their own distinctive ecosystems and varying plant communities. Thadagai hills (950 m above MSL) located in Western Ghats of Coimbatore district, Tamil Nadu,
India pocess humid climatic condition with rich biodiversity. A study has been carried out to know the diversity study of moist semi evergreen forests of
416
Thadagai Hills. The species richness of this forest is 42.
The important tree species of this vegetation are
Terminalia arjuna , T. bellerica , Gardenia gummifera ,
Combretum ovalifolium , Sterculia foetida , Syzygium cumini , Cordia obliqua , Carissa carandas and shrub species like Atalantia monophylla , Cissus vitiginea ,
Cayratia pedata , Gemelina arborea , Asclepias curssavica , Mundulea sericea etc. are also more common. The main species in herbaceous layer of the study area are, Ammania baccifera , Portulaca tuberosa ,
Ludwigia hyssopifolia , L. octavalis , L. peruviana ,
Catharanthus pusillus , Ocimum tenuiflorum etc.
However, utility value of these bioresources must be analysed to prepare conservation strategies.
P0179 – ePoster
Phenotypic plasticity leading to sympatric speciation in populations of
Ephedra
in Lahaul and Spiti (India)
Sharma, P 1 , Uniyal, Pl 1 , Hammer, O 2
1 Delhi University, India;
University of Oslo, Norway
2 Natural History Museum,
Phenotypic plasticity is likely to play a significant role in speciation. Despite sustained interest in the morphology of Ephedra , growing in extreme conditions in hot as well cold deserts in India, the structural and developmental bases of evolution highlighting phenotypic plasticity for transitions in vegetative and sexual systems and adaptive strategies leading to speciation are not investigated. Here, an exploration is made into the relative contribution of phenotypic plasticity to the phenotypic variations, identifying the cause of speciation in populations of ethnobotanically, medicinally, ecologically and evolutionary significant genus Ephedra . Phenotypic plasticity of qualitative and quantitative traits
(morphology of stem, scale leaf, male and female strobilus) have been investigated in ten populations of the shrub Ephedra (Ephedraceae), native to Western
Himalaya, in the Lahaul and Spiti region (altitude up to
3800 m), using a combination of field and laboratory measurements including Principal Coordinates Analysis
(PCoA) for ordination. All data analysis was carried out in the software Past, version 1.98. An altitudinal morphocline (Spearman’s rank order correlation rs =
0.95, p < 0.001) and phenotypic plasticity is evident. The samples from each locality form tight, clearly separated groups in this morphospace. Conservative and stable, reproductive characters also highlight three new species
( E. sumlingensis sp. nov., E. kardangensis sp. nov., and
E. khurikensis sp. nov.). First report of Phenotypic plasticity as a major evolutionary force in populations of
Ephedra via habitat adaptation through the variability of various morphological characters causing sympatric speciation is revealed. Efficient conservation of the populations can only be based on habitat management, to favour the maintenance of microenvironmental variation and the resulting strong phenotypic plasticity.

P0180 – ePoster
Ecological and palaeoecological understanding of mangrove wetlands of Maharashtra (west coast,
India) towards better planning and management
Shindikar, MR 1 , Gunale, VR 2 , Kumaran, KPN 3
1
2
College of Engineering, Pune, Maharashtra, India;
Dept of Botany, University of Pune, Maharashtra,
India;
India
3 Agharkar Research Institute, Pune, Maharashtra,
Mangrove, a highly specialized coastal wetland, along the Maharshtra coast (north west coast, India) is analyzed on the field based investigation in recent years. The present day mangroves and their relics represented in the coastal deposits have been explored to study their diversity and distribution with associated ecological and conservation issues. The status of forest diversity is studied in view of conservation of these imporatant coastal resources. Out of 68 sampling sites along the estuaries, creeks and other mangrove habitats, the locally sensitive areas or hot spots are identified considering the overall diversity and composition of mangroves. The 18 mangrove species show specific pattern in their distribution and density along the coast. Analysis of surface and subsurface sediments from these wetlands has provided insight for their antiquity during Holocene period. The techniques like palynology and environmental magnetism in the framework of C14 and
Luminescene dating are proved significant to know the ecolgical shifts in response to changing climatic conditions in the past. Further, current utilization pattern of these wetlands is understood from the secondary sources with the review of conservation efforts at different levels. The threats to mangroves are also studied towards its incarporation in an appropriate management practice. As such this information is significant to take a stock of how mangrove wetlands have locally responded to the coastal evolution and accompanying environmental changes in the past on the basis of their present diversity and composition. This could form a basis for the better planning of conservation and sustainable utilization of mangrove resources in
Maharashtra.
P0181 – ePoster
The distribution of
Cladonia
species at subalpine region of Mt Fuji in Japan
Shiori, I 1 , Susumu, O 1
1 Chiba University, Japan
Genus Cladonia communities occur near forest limit of
Mt.Fuji. These communities are rare in Japan. It is important to preserve it with knowledge containing lichen ecology. Little attention has been given to the ecology of lichen communities in Japan, yet. We investigated the relation of distribution of Cladonia communities to plants above lichens. Study site is Oniwa of Mt.Fuji at 2450 m high. In Oniwa, patchy vegetation appears surrounded by evergreen coniferous forest by
Abies vetchii and Tsuga diversifolia. The patches are 1–6 m wide and composed of Larix kaempferi , Fallopia japonica and lichens. We surveyed the distribution of
Cladonia communities within the patch using 4 transects per patch. Bryophytes occur under Larix kaempferi at the center of the patch.
Cladonia stellaris occurs around them. C. nigripes tends to occur also inside the patch. C. mitis occurs outside the patch. C. squamosa shows no clear pattern of distribution within the patch. C. stellaris ,
C. nigripes and C.
mitis are dominant lichens in the patch. Their individuals are larger than the other
Cladonia species. They make dense colonies.
Meanwhile, C. squamosa hardly dominates in the study site, and its individuals are smaller. They don’t make dense colonies. The distribution of Cladonia species making a dense colony shows the relations to distance from trees. C. stellaris and C. nigripes occur near trees within the patch. C. mitis occurs apart from the tree within the patch. These distribution patterns indicate that trees control their colonization. Plants reduce the wind speed. Because C. mitis grows well under windy condition, it can grow apart from the tree. It reduces the site inside the patch by competition with C. stellaris .
Because C. stellaris is sensitive to the wind among genus
Cladonia , it can’t grow outside the patch under windy condition far from tree. The distribution of Cladonia species colonizing is affected by plants. The distribution of C.
squamosa shows no clear pattern.
P0182 – ePoster
Land use, vegetation and climate change in Manaslu
Conservation Area, Nepal
Shrestha, K 1 , Scheidegger, C 2
1 Central Dept of Botany, Tribhuvan University,
Kathmandu, Nepal; 2 Swiss Federal Institute for Forest,
Snow and Landscape Research, Birmensdorff,
Switzerland
The central Himalayan region contains biodiversity reserves that are under increasing threats from stressors including shifting cultivation, nomadic grazing, stand fragmentation, and climate variability and change.
Climate change is likely to influence agricultural practices in traditional land-use systems. Climate and land use change, as well as their interactions will result in a dramatic reshuffling of species. Invasive species have been documented to spread into natural environments, often to the detriment of rare or endangered organisms.
The Manaslu Conservation Area (MCA) is Nepal’s newest conservation area established to balance biodiversity conservation and local livelihoods. The
MCA is bordered by Tibet Autonomous Region of China to the north and east, part of Gorkha district to the south, and Manang and Lamjung district to the west. The total area of the MCA is 1663 sq km, with a population of over 9000 people residing in 56 small villages of seven
Village Development Committees, and the vast majority of the population is ethnically Gurung. In the conservation area there are 15 peaks that are more than
6000 m above sea level, including the seventh highest peak of the world, Mt. Manaslu (8136 m). Attempts will be made to identify and analyze how different levels of land-use intensity (from natural forests, disturbed forests, meadows to arable fields) and climate do affect biodiversity in high-elevational landscapes in the southern slope of Nepal Himalaya. The general aim of
417

the study is to investigate the relationships between the biodiversity of vascular plants in the Nepal Himalaya below the timber line (2400 m to 4000 m) and the two main environmental drivers, i.e. land use and climate in different valleys of MCA. Biodiversity will be characterized by different measures in four land use types ranging from closed forests to open landscapes, and climate analyzes will include both temperature and precipitation. The analysis include consequences of biodiversity change for species richness of plants, plant functional groups; rare, endangered and endemic plant species, and selected ecosystem goods and services including high medicinal and aromatic plants. The results should thereafter allow to quantify the impact of land use and climate on vascular plant diversity and to estimate potential changes in this biodiversity under land use change and climate change scenarios. The outcomes of the research will be useful in the consequences for sustainable land-use and appropriate conservation measures to mitigate climate change effects in plant biodiversity in the Himalayas.
P0184 – ePoster
Structure, condition, botanical composition and current distribution of grasslands in the state of
Aguascalientes (Mexico)
Sierra, JC 1
A 2
, Siqueiros, MA 1
, Esparza, E 3
, Martínez, J 1 , Rodríguez,
1 Universidad Autónoma de Aguascalientes (UAA),
Mexico; 2 Instituto Nacional de Geografia y Estadística
3 (INEGI), Mexico; Comición Técnico Consultivo Para
La Determinación de Los Coeficientes de Agostadero
(COTECOCA), Mexico
Aguascalientes is a small political state located in central
Mexico, characterized mainly for arid and semiarid vegetation in most of its territory. One of the most important plant community is the arid grassland, which has been one of the most impacted by the activities of humans. In Aguascalientes, the grassland areas are located in the central plateau, where the majority of human settlements are located. The aim of this project, is to explore the general condition of the grasslands. We will evaluate the structure, type, botanical composition, current status, and distribution, as well to determine the degree of disturbance that these communities are facing.
The project is carried out for the UAA (Universidad
Autónoma de Aguascalientes), in collaboration with two government institutions (INEGI and COTECOCA). It includes two parts, one focusing on image interpretation and GIS modeling information techniques. The other one, centers in field sampling and in situ techniques, using the Canfield line as the sampling method. We planned 180 sample points to cover all Aguascalientes grasslands. So far, we have in advance of 40% sampled.
We found 49 families, 199 species, and have characterized 10 types of grassland, most of them with a high degree of disturbance.
P0185 – ePoster
Conservation of state park 'Cerro El Faro'
Silva Torres, B 1 , Gonzalez Ibarra, M 1
418
1 Universidad Autonoma Metropolitana, Mexico
The State Park 'Cerro el Faro' is a temperate forest made up of different species of Pinus , Quercus and Cupressus .
This park has been subjected to intense timber exploitation to supply a neighboring paper factory. Since
2005 an agreement was signed to allow the UAM to administer it for 25 years. The objective of this restoration process is to recover not only forest mass from a quantitative point of view but also to insure the forest quality allowing for the recovery of environmental goods, services and processes. It is important for the recovery of trees but is also necessary for the recovery of the soil, of medicinal plants which have local use, the fauna, etc. First a diagnosis was made of the zone and on that base a Management and Conservation Program was designed where restoration activities were included. The restoration activities included: plague elimination for the bark beetles, the rehabilitation of the neighboring trees, the elimination of weeds and a determination of the fire index. This paper shows the results obtained since 2005.
P0187 – ePoster
Hosts, habitats and light: surveys of native and exotic vines in the Sydney and south coast regions of
1
Australia
Smith, L 1 , Robinson, SA 1 , French, K 1
School of Biological Sciences, University of
Wollongong, NSW, Australia
Vines (climbing plants) have been described as
‘structural parasites’ due to their use of other plants for support, and the subsequent structural and physiological stress that this causes vine hosts. Many invasive climbers or scramblers are recognised as having serious detrimental effects on the communities and ecosystems they invade, however the physiological effects of vine invasion are seldom emphasised. In addition, native vines
(particularly woody vines or lianas) can also impair host growth and reproduction, and may cause just as much structural damage as invasive vines. Some species may be more susceptible to infestation by certain vines due to morphological traits. Alternatively, hosts may be locally abundant in microclimates preferred by vines. Vine growth (and therefore ability to impact hosts) depends upon the availability of light and support, which may both vary between the edges and interiors of forest fragments. There is currently a lack of information on whether some species of native plant are more likely to be used as hosts by both native and exotic vines, particularly in Australia. Such information is required for successful management of invasive vines. We aimed to determine whether the distribution of exotic and native vines differs between edges and interiors of forests, and whether some species of vines preferentially grow on certain trees or shrubs. Three distinct habitat types in the
Sydney and south coast regions of New South Wales,
Australia, were surveyed in August to November 2010.
Each vine species present was identified on ten hosts on the edges and interiors of forest fragments. Sixteen exotic and 26 native vine species from 21 families were recorded. The composition of vine communities differed between habitats but not between edges and interiors, possibly because these locations did not vary

significantly in terms of light or other abiotic factors. The two most common exotic vines were Araujia sericifera and Delairea odorata , with Stephania japonica and
Cissus antarctica being two of the most abundant native species. At present, a controlled experiment is underway to investigate the effect of light on competition between these vines and a native host, Acmena smithii . Of the seven most abundant vine species in this survey, five showed preferences for certain host species. In contrast,
A. sericifera was not associated with any species of host, and thus has the potential to exploit (and possibly damage) a wide variety of native species. The results of these investigations into vine preferences and impacts may aid future efforts to prioritise the control and eradication of invasive vines in south-east Australia.
P0188 – ePoster
National eco-regional planning needs to implement the global strategy for plant conservation
Squeo, FA
Martinez, K
1,2,3 , Stoll, A
4
4 , Gaymer, C F 4 , Letelier, LE 4 ,
1 Universidad de La Serena, Chile;
& Biodiversity (IEB); 3
2 Institute of Ecology
Center for Advanced Studies In
Arid Zones (CEAZA), Chile; 4 Universidad Catolica del
Norte, Chile
Within the framework of the Convention of Biodiversity
(CBD), Republic of Chile agreed to protect 10% of terrestrial and marine ecosystems by 2010. Gap analysis for biodiversity conservation was performed. As elements of biodiversity were considered the eco-regions, habitats, communities, services ecosystem and threatened species. As conservation goal used 10% of the surface, except for the threatened species (from 25% to 75%).
Considering only public protected areas, Chile still does not comply with the goal to protect 10% of the terrestrial or marine eco-regions and presents empty conservation most objects conservation. Only seven of the 13 terrestrial eco-regions met the goal and none of the eight marine eco-regions. In this scenario, Chile can demonstrate that it has complied with two-thirds of the terrestrial goal and only 0.3% of the themarine target, and in total (i.e., terrestrial and marine) only 11.8%.
Private protected areas increase terrestrial protected surface by only 3.3% (moving from 32.3% to 29.0% the unfulfilled goals). Conservation portfolio generated with
MARXAN 2.1.1 for different scenarios indicate that in
Chile between 18.5% to 21.3% of the national surface is required to achieve the conservation goals of 1433 considered objects (near 50% of the Earth's surface and ca. 13% of the marina). The terrestrial surface required decreased to 37.5% if goals for terrestrial animals are not considered. We considered that the use of plant communities is a novel approach to include most of the conservation objects. Eco-regional planning is a dynamic process, which improves to the extent that better quality information is available and is adjusted in the course of its implementation.
P0189 – ePoster
Procedure for single-pollen genotyping in ecological research
1
Suyama, Y 1
Tohoku University, Japan
Direct determination of multilocus genotypes and DNA sequences from single pollen grains is an efficient method and can be used to answer questions in different research areas such as pollination ecology and genomics.
For example, in plant population analyses, this technique will allow the estimation of pollen flow by directly tracking the actual movement of individual pollen grains.
Further, in genomic studies, this technique will enable direct analysis of the haploid DNA sequence of single pollen grains. The essential prerequisites to perform this technique are (1) the ability to manipulate small objects in small amounts of liquid by using standard pipettes, (2) determine optimal conditions to minimize the loss of
DNA, (3) identify the optimal conditions for polymerase chain reaction (PCR) amplification using extremely lowcopy template, and (4) eliminate possible contaminants.
Owing to its simple yet efficient approach, the singlepollen genotyping technique has become the technique of choice among molecular ecologists. This technique is expected to be used extensively in various disciplines in the future. I will present a comprehensive description of the laboratory protocol for this technique with some application studies.
P0190 – ePoster
Phytosociology and successional characterization in a flooded area of Iguaçu National Park - PR, Brazil
Temponi, LG 1 , Cardozo, AP 1
1 Unioeste, Cascavel, Brazil
Floristic and phytosociological surveys aim to inventory an association between species among the flora studied, and aim to contribute to the knowledge of local flora, being essential for preservation. Iguaçu National Park
(ParNa Iguaçu) is the largest remnant of the Interior
Atlantic Forest of Brazil. The aim of this study was to conduct a floristic and phytosociological survey in a flooded area of the ParNa Iguaçu, to gain an understanding of the flora of the park, and to support recovery actions of similar areas. 18 plots were demarcated, 10 x 10 m in size, totaling 0.18 ha of sampling area. Individual trees with a circumference at breast height (CBH) of 15 cm or larger were included in the sample. The specimens were deposited at the Western
Paraná State University Herbarium, analyzed, and identified. For the phytosociological analyses, FITOPAC
1.6 was used. The sample included 186 individuals from
19 families, 33 genera, and 35 species. The Shannon-
Wiener (H') diversity index was estimated at 3.085 for species and 2.493 for families, with Fabaceae having the highest species richness. Of the species sampled, 10 are recommended for restoration of degraded areas in the region, 54.83% being cited for flood areas. A predominance of secondary species can be seen, indicating that the area is at an intermediate stage of regeneration.
419

P0191 – ePoster
Metapopulation dynamics of the Mistletoe
Psittacanthus robustus
Mart. (Loranthaceae) in savannas areas
, Guimarães, CDC 2 , Coelho, GO 3 , van den Teodoro, G 1
1
Berg, E 4
2
University of Campinas - UNICAMP, Campinas, Brazil;
Federal University of São João Del Rei - UFSJ, São
João del Rey, Brazil;
UFLA, Lavras, Brazil;
Lavras, Brazil
3 Federal University of Lavras -
4 Federal University of Lavras,
The mistletoes’ habitat presents a peculiar spatial structure. It is composed by a collection of patches that can potentially be colonized in an inhospitable matrix formed by open space and trees not susceptive for colonization. In this way, the mistletoe populations are constituted by local sub-populations can be considered a metapopulation. The aim of this study was to evaluate the dynamic of mistletoe Psittacanthus robustus Mart. and its host Vochysia thyrsoidea Pohl and investigate if the P. robustus dynamics can be a metapopulacional model. It was carried out in three savanna areas in Minas Gerais state, Brazil, two of them classified as 'Outcrops
Savanna' and one as 'Sensu Stricto Savanna'. The first area is located in Parque Ecológico Quedas do Rio
Bonito (PEQRB 21º19'45.31''S e 44º58' 22.69''W), the second in Lavras and third one is located in the municipality of Carrancas (CZ 21º28’16”S and
44º37’21”W and CE 21º27’59”S and 44º42’10”W). 2.8 hectares were inventoried in each study area. The populations of P. robustus and V. thyrsoidea were monitored in an interval of six months. In site PEQRB, six consecutive surveys were carried out and in sites CE e CZ, three surveys were conducted. The rates of growth, colonization, re-colonization and extinction of the mistletoe were evaluated in each habitat patch (individual
V. thyrsoidea ). The change rates were low for the host and higher for the mistletoe. Metapopulation parameters showed that there was a decrease in the proportion of patches occupied in the study period. In the site PEQRB, the local sub-population extinction was due to the patch habitat extinction. The parasitism intensity can lead the host death; in this site all the patch habitat extinction had high number of P. robustus . In site CE, at the 2nd inventory, the sub-populations extinction was larger than the patch colonization, however, there wasn’t extinction of habitat patch and only the mistletoe sub-populations became extinct due to fire. In area CZ, the largest decrease of proportions of patches occupied, occurred at the 2nd inventory. In this site all extinction events of P. robustus sub-populations occurred because of a fire event between the first and second inventory. In conclusion, the
P. robustus dynamics can be viewed as a metapopulation, but each site presented its own dynamics due to the different fire occurrence. The P. robustus is a fire sensitive species and the burning controlled the mistletoe population size in CE and CZ sites. In these sites only the local sub-population extinction occurred, and patch extinction didn’t occur. However, in the site PEQRB
(without fire) the mistletoe presented a high local population and this caused patch extinction.
420
P0192 – ePoster
Correlations among stem and leaf traits in six rockoutcrop savanna species
Teodoro, GS 1 , Muler, AL 1 , Oliveira, RS 1
1 University of Campinas – UNICAMP, Brazil
Plant functional diversity can be assessed by identifying major axis of trait variation. Trade-offs among leaf functional traits such as leaf mass per area (LMA) and maximum photosynthesis (Amax) have been broadly described across plant communities and reflect plant resource use strategies. A 'wood economics spectrum' has been recently proposed, in which wood density (WD) and minimum water potential (WPmin) are considered key traits of stem hydraulic function. However, the tradeoff among these traits (WD and WPmin) has not been broadly tested and there is still no consensus whether leaf and stem traits are well correlated with each other. Our purpose was to investigate if there is a trade-off between stem traits (WD, WPmin) and leaf traits (LMA, Amax, stomatal conductance and water use effiency) in six dominant outcrop savannas species. The study was carried out in ‘Fazenda Vale do Céu’, Delfinópolis,
Minas Gerais state, Brazil, in a rock-outcrop savanna community, locally known as ‘Cerrado Rupestre’. This physiognomy occurs under a highly seasonal rainfall regime, over quartzite or sandstone outcrops and shallow soils with low water and nutrient availability. The study species were Byrsonima sp. (Malpighiaceae),
Eremanthus sp. (Asteraceae), Myrcia lingua (Myrtaceae),
Myrsine sp. (Myrsinaceae), Palicourea rigida
(Rubiaceae) and Qualea cordata (Vochysiaceae). Wood density was positively correlated with LMA (r²=0,1642; p=0,0494) and water use efficiency (A/gs) (r²=0,2797; p=0,0079). However, in contrast with previous studies, we found no correlation among wood density and minimum water potential, neither among LMA and
Amax. The correlation among two key stem and leaf traits (LMA and WD) supports the idea of whole-plant functional integration and may be informative for predicting plant performance under future environmental change.
P0193 – ePoster
Gametophyte ecology of the American Hart's-tongue
1 fern
Testo, W 1 , Watkins, Jr., J 1
Colgate University, New York, USA
This study investigated the gametophyte ecology and physiology of the American Hart’s-tongue fern,
Asplenium scolopendrium var. americanum , a federallylisted species native to eastern North America and found only in lime sinks and glacial plunge ravines with calcium-rich substrates. Over 90% of the approximately
4000 plants remaining in the United States are found in two counties in central New York, where populations have recently experienced a sharp decline. The goal of this work was to generate a better understanding of the ecology of the gametophyte generation and to investigate potential reasons for the species’ decline. Gametophytes

of this species were grown at 20 o C and 25 o C and in low-
(2.5g/L), medium-(5g/L), and high-(7.5g/L) calcium treatments, and developmental pattern and growth rates were recorded. Spore germination occurred after 11-15d, and was of the Vittaria -type. Germination rates were similar in all treatments, but gametophyte growth was significantly slowed in the 25 o C, remaining in the protonemal stage of development indefinitely. Calcium affected sexual development, with gametophytes grown in the high-calcium treatments developing gametangia earlier and having higher sporophyte recruitment rates
(13.9%) than those grown in medium-(2.3%) or lowcalcium (0.3%) treatments. Isolation experiments were conducted to investigate the possibility of intragametophytic selfing in the species. Self-fertilization did not occur, though several cases of apogamy were observed in the high- and medium-calcium treatments. A unique form of asexual reproduction was also discovered.
Gametophytes began to produce copious outgrowths along the gametophytic margin at ca. 90d. These outgrowths developed pluricellular meristems and eventually became independent gametophytes capable of further asexual reproduction. Additionally, several comparative developmental studies and ecophysiological experiments were conducted with A. scolopendrium and four common associate fern species: A. trichomanes ssp. quadrivalens , A. rhizophyllum , Cystopteris bulbifera , and
Dryopteris marginalis . Of these species, A. scolopendrium exhibited the lowest germination percentage after 60 days, and was also most sensitive to desiccation and freezing. Each of the other species studied appeared to be more tolerant of environmental changes, developing normally at both 20 ° C and 25 ° C and in all calcium treatments. These findings suggest that A. scolopendrium is particularly susceptible to the effects of climate change and habitat disturbance, and may be outcompeted by other species on account of its slow growth, limited reproductive potential, and extreme habitat specificity.
P0194 – ePoster
Genetic variation in photosynthetic response of wheat to elevated CO
2
in Australian grains free air carbon dioxide enrichment (AGFACE)
Thilakarathne, L 1
G 3 , Tausz, M 4
, Posch, S
, Seneweera, S
1 , Norton, R
1 , Borgeault, M
0
5
, Fitzgerald,
1 Dept of Agriculture and Food Systems, University of
Melbourne, Australia; 2 International Plant Nutrition
Institute, VIC, Australia; 3 Victorian Dept of Primary
Industries, VIC, Australia;
Ecosystem Science, University of Melbourne, Australia;
5 CSIRO Plant Industry, QLD, Australia
The present atmospheric CO
2
4 Dept of Forest and
concentration is rising and is predicted to double by the end of the 21st century and this is likely to have a profound effect on plant growth and yield, especially for C
3
plants. A large genetic variability in growth response to elevated CO
2
(eCO
2
) has been documented among different species and there is very limited understanding of the underlying physiological mechanism. In this study, wheat ( Triticum aestivum L.) was used as a model plant to investigate the physiological basis of the intraspecific variation in plant response to elevated CO
2
. Seven wheat ( Triticum , aestivum L.) cultivars having different physiological traits, Janz, Yitpi (high tillering), Silverstar, H45
(synchronous tillering), Drysdale, Hartog, and Zebu
(medium tillering) were grown in the Australian Grains
Free Air Carbon dioxide Enrichment (AGFACE) facility.
The effects of rising Carbon dioxide (ambient aCO
2 ppm, elevated eCO
2
~380
~550 ppm) on plant growth, photosynthesis (A) and its related processes were investigated. Gas exchange measurements of flag leaf blades were carried out at pre and post anthesis stages. A versus intercellular CO concentration (Ci) was used to
2 estimate apparent maximum carboxylation capacity
(Vc.max) and photosynthetic electron transport rate
(Jmax ). Immediately after gas exchange measurements, leaf blades were sampled and leaf nitrogen (N) concentration was measured. A large variation in A was observed among the cultivars tested when measured at growth CO and eCO
2
2
concentration (either aCO varied from 20–23 and 20–29 µmol CO
2
2
or eCO
2
m -2 respectively. The highest pre-anthesis A responsiveness (eCO
2
/aCO
2
s -1
). A
at aCO
2
) was 45% for Janz while
Yitpi showed the least stimulation. In contrast, post anthesis A was significantly decreased for all cultivars when compared to pre anthesis measurement where Yitpi and Hartog showed the least reduction. Mechanistic analysis of gas exchange data showed large variation in apparent maximum carboxylation capacity of Rubisco
(Vc.max) and photosynthetic electron transport rate
(Jmax). A reduction in Vc.max at eCO
2
was prominent at the post anthesis stage suggesting that strong A acclimation to eCO
2
occurs in the later part of leaf development. There were large genotypic differences in
A acclimation to elevated CO strongest acclimation (Vc.max at eCO aCO
2
). Leaf N concentration tends to decrease at elevated CO
2
2 where Zebu showed the
2
versus Vc.max at
and found a large genetic variability which was well correlated with Vc.max was found.
Acknowledgement Funding from the Australian
Commonwealth Department of Agriculture, Fisheries and
Forestry (DAFF) and the Grains Research and
Development Corporation (GRDC) s gratefully acknowledged.
P0196 – ePoster
Floristic, edaphic and climatic aspects of an island forest in a doline on quartzitic rock fields at Itacolomi
1
State Park, Brazil
Tonaco, A 1 , Baêta, H 1 , Leite, M 1 , Kozovits, A
Universidade Federal de Ouro Preto, Brazil
1
Rocky fields consist of a mosaic of plant communities distributed according to variations in microtopography, soil and microclimate characteristics. In the Itacolomi
State Park (ISP), in Ouro Preto, Minas Gerais, Brazil, part of the complex composing the rocky fields on quartzite is represented by dolines occurring as closed, circular or elliptical depressions and variable dimensions.
Doline is a kind of karstic geomorphologic feature and are uncommon on quartzitic rocks, but are widely distributed in the study region. This work aimed to know the florula of a doline and its relationship with the flora of other phytophysiognomies in ISP and to assess physical factors that influence the establishment of the plant community within the doline. The studied doline is
421

located at 1600 m elevation and has an elliptical edge, with about 0,05 ha and 20 m of depth. It presents tree and shrubby vegetation, differing strongly from the physiognomies of rock outcrops and plateaus in the surroundings, which are predominantly dominated by herbaceous plants. In this work, floristic survey of vascular plants within the doline was carried out for 12 months, and was compared with the flora from rocky fields and forests of ISP. Also, physical parameters (soil fertility and microclimate) were monthly measured in and outside the doline, in its surroundings. We found 89 species and 40 families so far. Asteraceae (10spp.),
Solanaceae (8spp.), Rubiaceae and Orchidaceae (6spp.) had the greatest number of species and 20 of these plants are still not determined. Considering the other 69 species,
26 are common to both the rocky fields and forest from
ISP, 18 are common only to the rocky fields, 11 only to the forests, and 14 of them are new registers, being known only in the doline. In the doline, 0,17 species per m² area were found, which is a similar value found in other studies on plant communities in rocky fields in ISP and even higher than those measured in forest phytophysiognomies. The higher value of richness per area in the doline might be explained by more favorable soil and microclimatic conditions for tree species, the influence of the surrounding flora and the presence of species that do not tolerate fire, a frequent event on rocky fields. A PCA was performed with physical data in order to test the main differences between the dolina and its surrounding environment. In rainy season (Oct/Apr) soil
N and P were the parameters that mostly differed between areas, followed by soil water content, Ca and relative air humidity. During the dry season (May/Sep) the main factor was soil water content, followed by soil
K, relative air humidity, soil N and Ca and organic matter. The doline flora has a high richness of species, increasing the floristic diversity of rocky fields at ISP.
These results confirm the determinant role of geomorphologic processes in karsts, creating different edaphic and microclimate conditions in dolines, allowing the development of a very different flora and plant physiognomy.
P0197 – ePoster
Patterns of genetic variation in a native shrub of the drylands of southeast México
Trujillo-Argueta, S 1 , del Castillo, R 1
1 Instituto Politécnico Nacional CIIDIR Oaxaca, Mexico
Malacomeles denticulata (formerly Amelanchier denticulata ), is a member of the Rosaceae family. It grows mainly on deteriorated areas in shrubs, tropical dry forest and oak and pine-oak forests along its distribution area from Central México to Guatemala. We consider that M. denticulata might be an ecological important species for recovering degraded areas since it might be highly genetically differentiated to withstand such arid and fragmented places. In order to characterize the genetic diversity of this species on the drylands of
Oaxaca State, México, we used single nucleotide polymorphisms (SNPs). Several assays were designed, one from M. denticulata and three more from closely related genera: Amelanchier and Peraphyllum . Sequences were analysed with the programs Repeat Masker and
422
Basic Local Alignment Search Tool (BLAST). Each assay included a pair of primers and a pair of VIC and
FAM fuorescent probes. Leaves were collected from eight populations from Central Valleys in the State of
Oaxaca, México and were analysed for SNPs. A real time
PCR was used. Parameters of within-population genetic diversity and among-population divergence (Fst) were calculated. Only positive results were obtained with the
SNP design assay from M. denticulata which yielded recognizable homozygotes and heterozygotes genotypes.
An Fst = 0.l98 value provides evidence of genetic differentiation among the eight studied populations. The inbreeding of individuals relative to their populations or fragments, Fis, was -0.282 and the total inbreeding Fit was -0.028. Negative values can be explain by the excess of heterocygous observed in the most degraded locality
(Yanhuitlán) where observed heterocygosity (Ho= 0.44) was higher than the expected heterocygosity (He= 0.34).
Since heterozygous individuals were only detected in the most fragmented and degraded population, heterosis probably is an important mechanism to survive for
Malacomeles denticulata on this deteriorated area. These results show that high genetic diversity can be found in some populations following disturbance.
P0198 – ePoster
Phenology of the Western Australian endemic
Persoonia longifolia
(Proteaceae), a highly recalcitrant species required for minesite restoration
Chia, K 1, 2 , Turner, S 1 , Koch, J 3 , Dixon, K 1
1 University of Western Australia; 2 Kings Park and
Botanic Gardens, Perth, Western Australia; 3 Alcoa of
Western Australia
Persoonia longifolia is a mid-storey climax tree species from the jarrah forest of Western Australia. Whilst relatively common in natural bushland (between 350-440 plants per hectare), its regeneration on rehabilitated mine sites in Boddington and Dwellingup is very poor, with only 0 and 40 plants per hectare recorded several years after restoration work. In mining restoration the principle source of new recruits is from respread topsoil, as propagation from seeds is currently impossible due to complex seed dormancy. The use of cuttings for propagation has also proven to be problematic. As with other Persoonia species, P. longifolia stems are also harvested from wild plants for the export and domestic flower market as they are used in cut flower arrangements. There is significant horticultural interest in this species due to it graceful symmetry, textured bark and relatively small compact stature. Nevertheless, as found in restoration programs, propagation of P. longifolia for the cut flower trade and horticultural industry has also proven to be exceptionally difficult.
Despite being a common species within the jarrah forest environment, very little is currently known about the overall ecology of this species and in particular its reproductive biology. This study aims to address the lack of information concerning P. longifolia through investigating its phenology with the intent of developing a better understanding of its ecology in order to develop effective propagation approaches for this beautiful tree species. As part of this study, trees in three different geographical regions were monitored on a monthly basis

to measure vegetative growth, presence/absence of flowers, buds and fruits. Fruit counts were also made to determine peak fruiting season. Germination events, which are rarely observed under natural conditions, were recorded and seedlings monitored for survival. Trees in different areas were also subjected to different fire regimes and monitored to understand the impact of fire on the growth, fruiting and flowering, and germination events of this species. Exclusion cages were established over seeds placed out in the field to monitor germination events from seeds of known age and the impact of predation on these germinants. These experiments are intended to be a part of a longer-term study and will continue over a 3-year period to document changes in seed production and germination events in association with climatic variation over several years.
P0199 – ePoster
Determinants of geographic range size in the plant genus
Brachyotum
(Melastomataceae)
Ulloa, C 1 , Jiménez, I 1 , Bogler, D 1
1 Missouri Botanical Garden, Saint Louis, Missouri, USA
Patterns of range size in anyone group of organisms may best be explained by a combination of hypotheses but few studies have simultaneously considered more than two of them to assess their relative importance. We estimate the relative importance of five hypotheses as explanations of variation in geographic range size among species in the Andean plant genus Brachyotum
(Melastomataceae). Comparative studies suggest that general ecological and evolutionary factors may largely determine variation in geographic range size in a wide variety of taxa. These factors are differentially stressed by several currently prominent, non-exclusive, hypotheses. Two of these hypotheses relate to how characteristics of ecological niches determine the amount suitable habitat under contemporary environmental conditions. The niche width hypothesis proposes that the range of environmental conditions in which a species is able to survive and reproduce is a major determinant of geographic range size, so that there is a positive interspecific relationship between the niche width and geographic range size. The niche position hypothesis poses that geographic range size is determined by the spatial frequency of the environmental conditions in which a species is able to survive and reproduce. Two other hypotheses relate to dispersal limitation. The postglacial dispersal limitation hypothesis proposes that species geographic ranges have undergone major shifts since the last glacial maximum, as species responded individualistically to climate change by tracking suitable habitat on geographic space. On the other hand, the habitat-connectivity dispersal limitation hypothesis stresses the effect that the contemporary spatial arrangement of suitable habitat may have on dispersal and thus the proportional occupation of suitable habitat by individuals of a given species. Finally, the age-range hypothesis, pulls together several models about how the age of a species relates to geographic range size, reflecting a potentially general set of processes that determine the transformation of geographic ranges between speciation and extinction. Explaining interspecific variation in the size of geographic distributions is a central concern in ecology and biogeography, and studies in this field are needed to estimate the relative importance of different currently prominent hypotheses. The proposed research takes on this task using species of the Andean plant genus
Brachyotum as a study group, ideally suited for this purpose because they are highly detectable during field surveys and easy to collect. It integrates concepts from several areas in ecology and evolutionary biology as well as a wide array of approaches including models to estimate occupancy while account for detectability, ecological niche modeling, spatial analyses from landscape ecology, molecular systematics, and structural equation models. The size of the geographic range of a species is a fundamental ecological and evolutionary trait, and understanding its variation has far-reaching pure and applied biological significance. By example, geographic range size is thought to determine the likelihood of species extinction and species responses to future climate change. Thus, understanding the determinants of geographic range size leads to understanding extinction risk and species responses to future climate change.
P0200 – ePoster
Documentation of endemic plants of Nilgiris, southern
Western Ghats, India
Ramachandran, VS 1
1 Dept of Botany, Bharathiar University, Tamil Nadu,
India
The Nilgiris is a biodiversity rich hotspot due to its location where the Eastern Ghats joins with the Western
Ghats which attracts at most interests of biologists to work in. The level of endemism in the flora and fauna of any bio-geographical region determines its uniqueness.
The Western Ghats contribute more than 5% of species to the global pool. The Nilgiris has more than 2611 species of angiosperms which is more than of the British flora, of which 115 species endemic to Nilgiri Biosphere
Reserve. Working on the inventorisation and documentation of endemic plants in Nilgiris resulted in the collection of 40 species; in which habitat preference of these endemic plants were analysed and found that 23 species occur in the upper plateau (> 2000 above msl) of the Nilgiris; whereas 17 species prefer the habitat which is less than 2000 m. as the niche choice. The IUCN red listed plants collected from Nilgiris are Ixora lawsonii
Gamble, Mellicope indica Wight, Psychotria globicephala Gamble, Sophora wightii Baker, Symplocos oligandra Bedd., Youngia nilgiriensis Babcock,
Euonymus serratifolius Bedd. as endangered category.
The Nilgiris have experienced significant land use changes over the years and loses still occur in the extent of dense forests. This has affected the population of many endemics due to the reduction in the range thereby leading to local extinctions. Due to dam construction, hydroelectric projects, conversion of forest lands for agricultural purposes, grazing, invasion of weeds (alien species) and fire has led to habitat destruction resulting in loss of native flora. Surveys of the population and studies on the habitat requirements and the availability of suitable habitats are essential for conservation and management.
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P0204 – ePoster
Plant ecological studies of Kalsubai-
Harishchandragarh Wildlife Sanctuary (India)
Waman, M 1 , Mahajan, D 2 , Shindikar, M 3
1 Board of College and University Development,
University of Pune, India;
College,Pune, India; 3
2 Dept of Botany, Waghire
Dept of Applied Sciences,
Government College of Engineering, India
The present work represents ecological observations on
Kalsubai-Harishchandragarh Wildlife Sanctuary of
Ahmednagar district (India). In spite of biotic interference, the region is still rich in diversity. The present investigation was undertaken, with a view to bring out comprehensive ecological and other related vegetation aspects of Kalsubai-Harishchandragarh
Wildlife Sanctuary which has been declared as Wildlife
Sanctuary since February 25th, 1986. Total area of
Kalsubai-Harishchandragarh Wildlife Sanctuary is
36171.03 hectares. Out of the total area, 17119.86 hectares was forest, 2884.17 hectares protected forest and
16167 hectares non-forest area. Before declaration of wild life sanctuary, this area was a part of Rajur forest range of Ahmednagar Forest Division (India). The semievergreen and moist, mixed deciduous forest of the study area consisted of a wide range of economically important plant species. The soils were reddish-brown and chiefly lateritic and gravely along the slopes. The floristic evaluation of plants has resulted in enumeration of 629 species. They constitute 372 genera of 112 families. The ten most dominant families of the region on the basis of species number were Poaceae (69), Fabaceae (59),
Asteraceae (42), Acanthaceae (29), Cyperaceae (28),
Rubiaceae (26), Euphorbiaceae (21), Lamiaceae (20),
Asclepiadaceae (19) and Scrophulariaceae (16). The phenological observations were resulted in documentation of 85 plant species, out of which 63 are trees, 4 Climbers and 18 shrubs. The physiognomy of this region was observed to be semi-evergreen to mixed moist deciduous. The top canopy was enclosed by
Mangifera indica , Terminalia bellerica and Ficus racemosa or Syzygium cumini , Terminalia chebula ,
Tectona grandis , Mangifera indica , and Olea dioica .
Some of the individuals attain the height of about 20 or more meters and spread horizontally. The second storey was occupied by Syzygium cumuni , Memecylon umbellatum and Phyllanthus emblica or Mallotus philippensis and Memecylon umbellatum . The species like Carvia callosa , Carissa congesta and Memecylon umbellatum comprise understorey of vegetation cover.
Due to increased practices of shifting cultivation the ecosystem was disturbed and moving towards the instability. As long as the tribal depend on this practice, they will not be able to improve. Shifting cultivation was one of the reasons of deforestation and also an important cause for soil degradation. Sacred groves were unique characteristics features of this wildlife sanctuary. They were rich in plant diversity and represent the index of natural vegetation. During the present study three new
(yet unnoticed) sacred groves have been investigated.
These were Ghatandevi (Ghatghar), Maruti-ban
(Koltemba) and Raghoba-ban (Pachnai). These were well preserved in their natural conditions and shows rich biodiversity. However, these need immediate attention from conservation point of view.
P0205 – ePoster
The impact of extremely cold weather on the first flowering dates of garden woody plants under
1 warming climate in Central Shanxi, China
Wang, L 1 , Zhang, Y 2 , Li, F 3
Forestry College, Shanxi Agricultural Unniversity,
China; 2 School of Landscape Architecture, Beijing
Forestry University, Beijing, China; 3 Economics and
Trade College, Shanxi Agricultural University, China
The weather from November 2009 to April 2010 was extremely cold in Shanxi, China, while the overall climate has been warming over the past 50 years, especially since the 1990s in the region. The impact of this period of extremely low temperature on the first flowering dates of garden woody plants was studied in
Taigu located in central Shanxi, China. The results showed: (1) the first flowering dates of all the woody plants were delayed significantly, with an average of 12 days delayed; (2) the first flowering dates were affected more for the species with earlier flowering dates than those with later ones.
P0206 – ePoster
Decomposition characteristic of leaf litters for main
1 forests at protective greenbelt in Karamay
Wang, Y 1 , Yin, LK 1 , Zheng, L 2
Xinjiang Institute For Ecology and Geography, Key
2
Laboratory of Biogeography and Bioresource In Arid
Land, Chinese Academy of Sciences, Beijing, China;
Graduate School of Chinese Academy of Sciences,
Beijing, China
According to the decomposition study of leaf litters for seven forests at protective greenbelt in Karamay, the results indicate that trend of leaf litters decomposition rate was 'quick- slow- quick', the order of it was
Elaeagnus oxycarpa and Populus alba var. pyramidalis +
Amorpha fruticosa > Populus euphratica and Amorpha fruticosa > mixed herbage, Populus alba var. pyramidalis and Ulmus pumila after one year decomposition. After the mixture of P. alba var. pyramidalis and A. fruticosa , it could significantly accelerate the decomposition of leaf litters. Litter decomposition processes were simulated using Olson's exponential models, which were found the best of fits to the data. According to the models, it needed 2.25-8.98 years to reach the half decay rate and 9.82-45.53 years to reach the 95% of decay rate for the seven forests in the experimental region, and the time was 7.17 and 7.88 times for the subtropical forest, and that of 2.66 and 3.04 times for the cold temperate forest. Nutrients content changed at the end of the experiment, the content of nitrogen and calcium was higher than initial value, the content of kalium and magnesium was lower than initial value, there was a little variation with the content of phosphor after one year later. Decay of organic carbon was fast and C/N value descended gradually. Kalium had
424

the most release rate, the next was phosphor, magnesium and calcium. Nitrogen was the net accumulation.
Cellulose content of P. alba var. pyramidalis , U. pumila and mixed herbage were higher than initial value, that of others was lower than initial value. Lignin content was higher than initial value except that of U. pumila . The trend of amount of bacterium was more in spring and autumn, less in summer, the amount of actinomyces and fungi was more and more with decomposition.
Correlation coefficient and probability value of decomposition rate with phosphor content, lignin content, C/P, lignin/N were higher and higher, but that with C/N was lower and lower, with decomposition.
Statistics analysis indicates that there were no correlations about decomposition of leaf litters with above indexes, and suggested that to establish comprehensive indexes correlation relation between litter substrate quality and decomposition.
P0207 – ePoster
Types and characteristics of vegetation in Urumqi
City
Yin, LK 1
Zan, Q 3
, Nan, WJ 2 , Jiang, FQ 1 , Yan, C 1 , Wang, L 1 ,
1 Xinjiang Ecology and Geography Research Institute,
Key Laboratory of Biogeography and Bioresource In
Arid Land, Chinese Academy of Sciences, Urumqi,
China; 2 Ollege of Agriculture and Gardening, Xinjiang
Agricultural University, Urumqi, China;
Limited, Hong Kong
3 Earthasia
According to the degree of human disturbance, community function and advantages of the dominant and constructive species, the paper divided the Urumqi urban vegetation into three parts: natural vegetation, half natural vegetation and artificial vegetation including three vegetation races, 13 vegetation groups and 125 vegetation types. Natural vegetation race was classified as 7 vegetation groups, i.e., coniferous forest vegetation group, broadleaf forest vegetation group, scrub vegetation group, desert vegetation group, meadow vegetation group, steppe vegetation group and alpine vegetation group with 82 vegetation types. Half natural vegetation race was divided into 4 vegetation groups, i.e., broadleaf forest vegetation group, scrub vegetation group, herbaceous vegetation group and partner person vegetation group with 33 vegetation types. Artificial vegetation race was classified as 2 vegetation groups, i.e., green space and agricultural vegetation group with 10 vegetation types. Urumqi had a full spectrum of natural vegetation race and rich type diversity. Dominated by the inland desert climate, the zonal desert vegetation types developed well, xerophytes scrubs and super-xerophytes scrubs, semi-scrubs and perennial herbaceous vegetation had obvious advantage. Due to human disturbance, the number of plant community species of the half natural vegetation race was increased significantly, and the proportion of trees and scrubs was increased when compared with the natural vegetation. In the community, the majority of the upper species of trees and scrubs were cultivated species, while mostly native herb group were composed of the wild and exotic species. Several species appeared a patchy distribution and linear distribution along the irrigation zone. All of these made the species diversity index of the half natural vegetation relatively higher. Green space of artificial vegetation race had a higher degree of fragmentation, showing spatial distribution of points, blocks, slabs and other forms in the center of the city, with a low coverage and little correlation between communities and populations.
Among them, the protection green space was dominated by a higher proportion of the introduced exotic species and naturalized species. Affected by the market economy, agricultural vegetation group was composed of three main vegetation types, i.e., food, vegetables and oilseeds.
P0208 – ePoster
Is Africa losing its umbrella?
Webber, BL 1, 2
C 5 , McKey, D 1
, Born, C 3 , Chandler, RE 4 , Djieto-Lordon,
1 CNRS, Montpellier, France;
Sciences, Wembley, Australia; 3
4
2 CSIRO Ecosystem
Stellenbosch University,
Matieland, South Africa;
UK; 5
University College London,
Faculté des Sciences, Yaoundé, Cameroon, Central
Africa
In the late 18th and early 19th centuries, colonial botanic gardens were staging posts for the global movement of a great diversity of plants within and between continents.
Some of these botanic gardens were neglected for long periods following their establishment as their purpose drifted between horticultural exchange or agriculture research, and between changes in management regimes with different interests and priorities. Many botanical garden plant introductions are now significant invasion problems and history reveals that plants were frequently re-introduced many times, even when in hind-sight it was clear that they had invasive properties. The Neotropical tree Cecropia has been moved between botanical gardens, either deliberately or accidently, many times since the first known export from Jamaica in 1778.
Although listed as one of the world’s 100 most invasive species and with key traits that facilitate rapid dispersal and colonisation, there is very little published research on the invasiveness of Cecropia , or the impact of these invasions on native biodiversity. In Cameroon we investigated the spread of Cecropia peltata from the
Limbe Botanic Gardens, where it was first introduced, toward Korup National Park, a largely primary stand of one of Africa’s most diverse lowland rainforests. In particular, we (i) document the competitive replacement of the native congener, Musanga cecropioides (the oldworld sister taxa to Cecropia ), by comparing present day distributions to a survey completed 20 years ago, (ii) investigate the effect of vegetation type on the relative density of the two taxa, and (iii) explore the implications of our findings for other countries where Cecropia invasions have escaped from garden introduction points.
P0209 – ePoster
Targeted taxonomy in Western Australia: positives and pitfalls
Wege, JA 1 , Shepherd, KA 1 , Butcher, R 1 , Thiele, KR 1
425

1 Western Australian Herbarium, Dept of Environment and Conservation, Australia
Western Australia has a rich and highly endemic flora which comprises over 12,150 native vascular plants, about a third of which are threatened or listed as having conservation significance. This inventory currently includes 1600 informally named taxa, the majority of which lack descriptive information. These undescribed taxa are often under-collected and poorly understood, and in the absence of accessible diagnostic information can be very difficult to recognise. This taxonomic backlog therefore presents a major challenge to conservation management. Increased exploration and mining of iron ore in Western Australia has exacerbated these challenges, with new species continuing to be discovered through floristic surveys of mineral-rich habitats and the botanical evaluation of mineral leases. One approach taken by the State's Department of Environment and
Conservation to deal with the taxonomic backlog has been to fund targeted research to expedite the description of threatened and poorly known species, particularly those vulnerable to mining activities. In the past five years, this focused approach has led to the formal description of 120 new taxa, many published in a special issue of the Western Australian Herbarium's journal,
Nuytsia (Volume 17). The mobilisation of taxonomic and ecological information for these species has greatly enhanced on-ground conservation efforts. A greater understanding of the distribution and rarity of many of these taxa has also been achieved through improved herbarium specimen identification and field survey. This short-term approach raises a number of issues. On the positive side, it has focused attention on the need for taxonomy, promoted collaborations, and led to tangible conservation outcomes. On the negative side, it has necessitated small, species-focused papers rather than revisionary taxonomy, and there has been limited potential for building taxonomic capacity in specific groups, or for tackling intractable species complexes.
The lack of adequate long-term resourcing for taxonomic research remains of paramount concern to biodiversity conservation in Western Australia.
P0211 – ePoster
Can rapid evolution in response to an environmental change alter levels of gene flow in an annual plant?
Welt, RS 1 , Lalchan, R 1 , Litt, A 2 , Franks, SJ 1
1 Fordham University, Bronx, NY, USA; 2 New York
Botanical Garden, Bronx, USA
The effect of climate change on temperature and precipitation patterns across the globe is a major environmental concern and understanding how species are affected by climate change is of great importance as these changes are expected to continue into the future. As a strong selective force, climate change can promote evolution in natural populations. Recent research found an evolutionary change in flowering time in two natural populations of Brassica rapa (Field Mustard) in southern
California following a five-year drought (1999-2004).
These two populations rapidly evolved earlier and more synchronous flowering times in response to this climatic event. This greater overlap in flowering time corresponds
426 to decreased phenological, and potentially reproductive, isolation between the populations, which can affect levels of gene flow. To determine whether levels of gene flow have changed concurrent with this drought, we will examine the genetic structure of these populations using microsatellites. We hypothesize that gene flow between these two populations of Brassica rapa increased following an evolutionary response to a climatic event.
This research will be the first to evaluate gene flow across time using an ancestor-descendent comparison approach and will elucidate the potential for environmental changes to alter population structure.
Furthermore, these results can provide insight on the response of natural populations to such changes. An increase in gene flow following a drought, as predicted for this experiment, can have two opposing effects on these populations: increased local adaptation through the spread of adaptive alleles, or decreased local adaptation as non-adaptive alleles are shared; either of which can impact the potential for an adaptive response to an environmental change. Understanding the genetic basis of a species’ response to environmental changes can benefit the conservation of species in a changing world.
P0212 – ePoster
Cutting grass on desert islands: genetic structure of
GAB spring populations of
Gahnia trifida
Clarke, L 1 , Whalen, M 1 , Mackay, D 1
1 Flinders University, Adelaide, Australia
Gahnia trifida commonly occurs in temperate wetlands of southern Australia, however relict populations are also present on Great Artesian Basin springs near Lake Eyre in arid central Australia. We used microsatellites to compare the genetic structure of spring and coastal populations to assess levels of genetic diversity and connectivity between the fragmented spring habitat.
Consistent with the expected consequences of habitat fragmentation, GAB spring populations of Gahnia trifida show lower levels of genetic diversity and much higher inter-population differentiation (HE = 0.32 ± 0.05, FST =
0.52) than populations at coastal sites (HE = 0.49 ± 0.04,
FST=0.22). Our results indicate dispersal amongst spring populations is insufficient to counter-act the effect of genetic drift. Spring populations may have persisted for long periods of time, but may have limited evolutionary potential due to low levels of genetic diversity. Thus,
Gahnia trifida populations at GAB springs are likely to persist if conditions remain stable, but may be vulnerable to environmental change.
P0214 – ePoster
Analysis of the plastid genome genetic variation of
Euryale ferox
Salisb. (Nymphaeaceae) populations in
South Korea
Jang, JH 1 , Kim, JH 1 , Won, H 1
1 Daegu University, South Korea
Euryale ferox Salisb. (Nymphaeaceae) is a herbaceous aquatic species distributed from India to Russian Far
East, including China, Taiwan, Japan, and Korea.

Euryale ferox has large floating leaves up to 2 m in diameter, with prickles on both sides, unlike its close relative Victoria in northern South America having prickles only on lower surface. It is designated as an endangered species group II in South Korea, due to threats of destruction and reduction of habitats following development and reclamation of wetlands. To study the genetic variation of the E. ferox populations in South
Korea, we have collected 559 individuals of 37 populations and their plastid genome genetic variation was studied. Sequence variation of seven plastid regions
– psbA-trnH IGS, psbK-psbI IGS, atpF-atpH IGS, trnL intron, trnL-trnF IGS, rpoC1 gene, and rpoB gene – were examined for the representatives of 22 populations, of which the psbA-trnH IGS and the psbK-psbI IGS regions only showed variation. 16 bp long stem region and four bp long loop structure was detected from the psbA-trnH
IGS region, with TCTT or AAGA inversion variation in the loop region. A9/A10 repeat and A/G substitution was detected from the psbK-psbI IGS region. When all the
557 samples were sequenced for the psbK-psbI IGS regions, we obtained 311 (56.8%) of A9/A, 20 (3.6%) of
A9/G, and 217 (39.6%) of A10/G. Also, we obtained 406
TCTT type (76%) and 125 AAGA type (24%) from the psbA-trnH IGS region. These results altogether comprised 5 plastid haplotypes for E. ferox in South
Korea. Generally only one haplotype dominated a population, and population with mixed haplotypes was rare. These results suggest that (1) there are at least five maternal founders for the E. ferox populations in Korea,
(2) the lack of genetic diversity indicates that selfpollination is more common than cross-pollination
(which is also indicated by lack of chasmogamous flowers in the field), (3) further research using microsatellite markers is necessary to unveil genetic structure of E. ferox . Also, this information will be helpful in conserving and restoring E. ferox populations in South Korea.
P0215 – ePoster
Incongruence between genetic and morphological characters - lack of molecular variation between two
Korean endemic
Iris
species
Won, H 1 , Lee, J 2
1
2
Daegu University, Gyeongsan City, South Korea;
Green Plant Institute, South Korea
Iris koreana is a Korean endemic species first recognized by Takenoshin Nakai in 1914, distinct from I. minutoaurea . While I. minutoaurea is widespread from
North to South of Korea, I. koreana shows limited distribution in SW part of Korea. Due to the limited distribution and threat of population extinction, South
Korean government designated it as an endangered species level II. Also, another Korean endemic Iris species, I. odaesanensis , described by the late Youngno
Lee in 1974, was designated as an endangered species.
Fortunately, recent floristic surveys have discovered moderate number of new localities for I. odaesanensis , however no new was found for I. koreana . Due to similar morphology among the three Iris species, taxonomic reality of I. koreana has been questioned. To solve the problem, we have examined morphological characters and sequence variations of the three Iris species in Korea.
Morphologically, leaf width and texture, number of flowers per inflorescence, color and pattern of calyx, relative position of ovary with bracts, length of ovary stipe, length of calyx tube, etc. have clearly diagnosed three species. Sequence variation clearly distinguished I. odaesanensis from two other Iris species. However, I. koreana and I. minutoaurea show no variation in sequences of multiple plastid regions surpassing chloroplast LSC and IR boundaries, matK gene, and nuclear xdh gene and nrITS region sequences. This incongruence between the morphological and sequence characters may have been caused by recent and fast divergence of I. koreana from I. minutoaurea morphologically, with the lag of genetic divergence, following geographical isolation and founder effect.
P0218 – ePoster
Positive associations between the Cushion Plant
Arenaria polytrichoides
(Caryophyllaceae) and other alpine plant species increases with altitude in the
Sino-Himalayas
, Niu, Y 1 , Cavieres, L 2 , Sun, H 3
1
Yang, Y 1
Kunming Institute of Botany, Chinese Academy of
Sciences; 2 ECOBIOSIS, Departamento de Botanica,
Facultad de Ciencias Naturales y Oceanográficas,
Universidad de Concepción, Concepción, Chile, and
Instituto de Ecology Biodiversidad (IEB), Santiago,
Chile; 3 Kunming Institute of Botany, Chinese Academy of Sciences
It has been proposed that the facilitative effect of cushion plants tend to increase with elevation as a result of increases in environmental harshness. However, this hypothesis has not been tested in the Sino-Himalayan
Mountains. We evaluated the spatial association of several plant species with the Cushion Plant Arenaria polytrichoides (Caryophyllaceae) at two elevations (4500 and 4700 m elevations) in the study site and monitored temperature, moisture and nutritional status of soil and within cushion in Lakaka Pass on Baima Snow
Mountain, SW China. While 14 species grow more frequently associated to cushions at the higher elevation, at the lower site only 3 species were positively associated to cushions. Eleven of the species that occurred at both elevations changed their spatial association from neutral or negative with cushions at the lower site to positive at the higher elevation. Substrate temperatures were rather similar between cushions and bare ground areas. Internal water content of cushions maintained higher moisture than bare ground areas at both elevations. Soils under cushions containing significantly higher availability of nitrogen and potassium compared to open areas at the higher elevation. Our results show that facilitation by A. polytrichoides cushions increases with elevation in the
Sino-Himalayan region. This facilitation effect by A.
polytrichoides cushion is probably due to the improved nutrient conditions provided by cushion plants in the higher elevation, and these conditions probably permit increased plant recruitment, growth and survival.
P0220 – ePoster
Wood density - what are its underlying components and why does it vary with different climates?
427

Zieminska, K 1
Westoby, M 1
, Butler, D 1 , Gleason, S 1 , Wright, I 1 ,
1 Macquarie University, Sydney, Australia
Water availability and temperature are the two most influential climatic factors shaping vegetation structure worldwide. A key plant trait that varies with these environmental properties is wood density (dry mass per fresh volume [g/cm³]). On a global scale, species in drier environments tend to have higher density wood.
Although this trait is one of the most commonly measured plant characteristics in ecological and forestry studies, it is still rather unclear what exactly wood density tells us about plant functioning nor why it trends with climate. Presumably, the answers to these questions lie in the structural composition of wood. Wood consists of several types of tissues, the main ones being: (1) vessels which conduct water and nutrients; (2) fibres, which give mechanical support; and (3) parenchyma which transports and stores carbohydrates. Presumably, higher vessel and parenchyma fractions (filled with water, gas or cytoplasm) decrease wood density, whereas larger fibre fractions or fibres with thicker cell walls increase density. The fraction of wood occupied by vessel lumens (cavities) is particularly interesting because of potential for tradeoff between primary functions of wood; namely water conductance and mechanical support. For example, wood with a large proportion of vessel lumens may be effective in conducting water but mechanically weak. In this work we asked how anatomical structure changes along rainfall and temperature gradients, and how these shifts in anatomy relate to wood density. To address these questions we studied 24 species, six from each of four sites chosen to contrast in annual rainfall and temperature. Properties measured included wood density, vessel and fibre properties, and fractions of individual tissues. The overall goal was to develop and test functional hypotheses concerning the observed trends in anatomy and density. Preliminary results refer to twelve species from two Tasmanian sites with different rainfall but similar temperature regimes. Across these species wood density was more strongly determined by the density of matter outside vessel lumens (i.e. density of the non-lumen fraction) than by proportion of wood occupied by vessel lumens (the vessel lumen fraction).
This suggests that increases in wood density might not be simply related to conductance of water. Non-lumen fraction consists of vessel cell walls, axial parenchyma, rays, fibres and, in some species, tracheids. The density of all these tissues together varied most strongly with fibre fraction but the slope was shallow. Other tissue fractions had little influence on density variation. It is likely that the effect of fibre fraction on non-lumen density might be cancelled out by coordinated shifts in fibre cell wall thickness. For example, a species with a large fibre fraction but thin fibre walls may have similar wood density to a species with lower fibre fraction but thick fibre walls. The material properties and the physiological strategies of the two woods might be quite different. Ongoing work includes investigating the potential roles of fibre properties and various tissue fractions in determining wood density and its variations across sites.
428
P0221 – Poster
Contribution to the Red List of Pakistan: a case study of
Silene longisepala
(Caryophyllaceae)
Ali, H 1 , Qaiser, M 2
1 Center For Plant Conservation, University of Karachi,
Pakistan; 2 Federal Urdu University For Arts, Science and Technology, Islamabad, Pakistan
Silene longisepala Nasir (Caryophyllaceae) is endemic to
District Chitral Pakistan. This taxon was known from 2 localities (i.e. Torikhoo and Chitral proper). After three years of continuous field studies during 2005-2007, 24 new localities in lower Chitral were discovered. In view of its population size of 553 mature individuals, extent of occurrence of 3587km2 and area of occupancy of 104 km2, the taxon is placed under the category of endangered (EN). According to its population size and number of localities reported i.e. 608 mature individuals were found in 22 localities in 2005, 520 mature individuals were found in 17 localities in 2006 and 531 mature individuals were found in 17 localities in 2007.
The main threat posed to the degradation of the taxon is its habitat destruction. In 2 localities i.e., Megegram
Bakhtooli and Suratwet Rumboor, 25% and 30.7% decrease has been observed in its population size, respectively, caused by the construction of road in the core habitat. Soil erosion resulted from deforestation, grazing and agricultural land extensions are among the other main threats responsible for the reduction in population size. There is urgent need to develop species specific conservation strategies, followed by comprehensive conservation action plan and thereafter their implementation at ground root level on the national scale.
P0222 – Poster
Ecological pollination studies in
Ephedra foeminea and
Ephedra distachya
from Greece
Bolinder, K 1 , Ickert-Bond, SM 2 , Rydberg, A 1 , Rydin, C 1
1 Uppsala University, Evolutionary Biology Centre,
Norbyv.,Uppsala, Sweden; 2 University of Alaska, UA
Museum of North Herbarium and Institute of Arctic
Biology,Fairbanks, USA
Recent findings indicate that two major radiations have occurred during the evolutionary history of Ephedra
(Gnetales). The first radiation in the Early Cretaceous resulted in a comparatively large diversity of ephedroids including species that share uniquely derived features with extant Ephedra . A significant part of this diversity went extinct in the late Cretaceous, causing a bottleneck effect in Ephedra . It is noteworthy, that extant species show an extraordinarily low amount of genetic and structural diversity. A second radiation, which started presumably 30 million years ago, gave rise to the present-day diversity. What caused this second radiation?
Based on our current understanding of extant Ephedra diversity, a key morphological innovation or range expansion to a new latitudinal zone appears unsatisfactory as explanations. Another hypothesis concerns a possible shift in pollination biology. Biotic pollination and co-evolution with pollinators is one possible explanation for the evolutionary success of

angiosperms. However, biotic pollination also occurs in gnetalean gymnosperms, e.g., in Welwitschia and studied species of Gnetum . While Ephedra is most often referred to as wind pollinated, there are reports indicating a possible variation in pollination syndrome within the genus. We study pollination ecology in two distantly related species of Ephedra from Greece that differ in key morphological features of the male cones: E. foeminea and E. distachya . The experiment consists of two treatments: 1) exclude both wind and insects in order to discover if, in the absence of pollination, ovules might develop by apomixis; 2) exclude insects but allow abiotic transfer of pollen to the cones. Two sets of controls were also established: 1) hand pollinating for each of the two treatments to assess treatment effects on seed set; 2) natural pollination to assess treatment effects on pollination. Observations in 10 minutes intervals will be made to conclude if any insects show interest in the plants. The study is part of an interdisciplinary project on pollination biology in Ephedra .
P0223 – Poster
Spatial ecology of fungal communities associated with hair roots of
Woollsia punge ns (Styphelioideae)
Price, S 1 , Anderson, I C 1 , Cairney, J WG 1
1 Centre for Plants and the Environment, University of
Western Sydney, Australia
Several Ericaceae families form ericoid mycorrhizal
(ERM) associations, and these are regarded as fundamental to the success of Ericaceae plants in environments that are characterised by several edaphic stresses. While understanding of ERM fungal diversity and functioning has increased in recent years, we know little of the spatial patterns of ERM fungal communities.
To address this we have investigated the spatial structure of fungal communities associated with hair roots of
Woollsia pungens (Styphelioideae) at a range of spatial scales to determine whether spatial separation distance influences the composition of root associated fungi. W.
pungens plants were collected from eight study sites on a
500 km transect established along coastal New South
Wales, Australia. DNA was extracted directly from the hair roots and the rDNA internal transcribed spacer (ITS) regions were amplified by PCR using the fungal primers
ITS1-F and ITS4. The fungal communities were profiled using terminal restriction length polymorphism analysis
(T-RFLP) and compared using a range of spatial and multivariate statistical analyses. The composition of the fungal communities was found to differ with increasing geographical distance at the largest scales sampled (100 –
500 km), while spatial heterogeneity was not evident at intermediate scales (40 m – 25 km). The community structure also varied substantially among individual plants from within each site, and thus spatial patterns in the composition of ERM communities are also predicted at smaller scales (< 40 m) than were sampled in this study.
P0224 – Poster
Entomophily and scent mimicry in Newfoundland dung mosses (Splachnaceae)
1
Gaskett, A 1,2 , Marino, P 3 , Raguso, R 2
University of Auckland, New Zealand;
University, USA; 3 Memorial University, Canada
2 Cornell
Flowering plants are well-known for using colours and scents to attract and deceive animals, but mosses are perhaps the most ancient agents of plant-animal behavioural manipulation and sensory exploitation.
Mosses in the family Splachnaceae are unusual because some species grow only on animal remains such as carcasses, bones, dung, and old bird nests. How can
Splachnaceae mosses locate, colonise, and spread between such patchy and ephemeral habitats? Most mosses must rely on passive spore dispersal by water, but the Splachnaceae have several unique features suggesting they exploit insect vectors for spore dispersal: brightly coloured ornaments, adhesive yellow spores, and appalling odours reminiscent of rotting carcasses or faeces. We analysed signalling and insect attraction to three Splachnaceae mosses, Splachnum ampullaceum ,
Splachnum pensylvanicum , and Tetraplodon mnioides in
Newfoundland, Canada. We analysed the scents produced by the mosses and their potential odour models, moose and coyote dung, and experimentally tested insect attraction to synthetic odours associated with herbivore, omnivore and carnivore dung. Investigating such poorlyknown and ancient plant-insect interactions provides considerable insight into the origins and mechanisms of more recently evolved relationships including pollination and seed dispersal.
P0225 – Poster
Effect of growth temperature on carbon assimilation and water use efficiency in chenopod species with different types of photosynthesis
Koteyeva, N 1
Edwards, G 2
, Voznesenskaya, E 1 , Cousins, A 2 ,
2
1 V.L. Komarov Botanical Institute of RAS, Russia;
Washington State University, USA
The purpose of the study is to evaluate effects of climate change on performance of chenopods which grow in semi-arid regions, and which have different structural and biochemical forms of photosynthesis. Two C
4
Suaeda species with different Kranz anatomical types ( S. taxifolia and S.
eltonica ), two unique species with the C
4 pathway inside individual chlorenchyma cell ( S.
aralocaspica and intermediate (
Bienertia sinuspersici
Salsola divaricata
), one C
) and two C
3
3
–C
4
species
( Suaeda maritima , S. heterophylla ) were grown under different temperature conditions to simulate predicted climate change. Analysis of gas exchange data in C
4 plants from three different growth temperatures was conducted. Compared to moderate temperature, there was a significant decrease in photosynthesis rates per unit leaf area in plants grown under temperature extremes (low and high), except for S. taxifolia . With increasing growth temperature there was an increase in the light compensation point for photosynthesis, indicative of greater respiratory loss of CO
2
. However, plants grown under low temperature had earlier saturation of photosynthesis with increasing light, indicative of low temperature limitation on the capacity of enzymes in
429

carbon assimilation. The water use efficiency (WUE) in
C
4
and C
3
–C
4
intermediate plants is higher than in C
3
, and it decreased with increased growth temperature. This suggests that, with global warming, when potential for water loss is high, the C
4
species with their higher WUE will have an advantage. Analysis of the temperature response curves of photosynthesis showed species specific differences in acclimation in plants with different forms of photosynthesis. The highest potential for acclimation to growth temperature was observed in the single-cell C
4
species Bienertia sinuspersici .
Measurements of the CO
2 compensation point (
Γ
) were made by three methods, including on-line massspectroscopy. Under moderate growth temperature,
Γ values characteristic of photosynthetic types were obtained, which were very low in C species. However, in two C
4
4
and high in C
3
species and in the C
3
–C
4 intermediate grown under low temperature, there was an increase in
Γ
values towards C
3
-like photosynthesis. In the C
3
-C
4
intermediate this was shown to be due to increased respiratory losses (analyses by immunolocalization showed retention of protein compartmentation, characteristic of intermediates). Measurement of carbon isotope composition ( 13 C/ 12
C
3
–C
4
C) of biomass did not reveal significant changes in photosynthetic type in the C
4 species under different growth temperatures. The growth response of the species under different temperature regimes was measured by increase in branch biomass. At the biochemical level, western blots are being analyzed to determine the quantity of photosynthetic enzymes
(PPDK, PEPC, NAD-ME, LSU Rubisco) in C
4
plants from different growth conditions. Current results indicate plants having the single-cell type of C photosynthesis do
4 not show marked differences when compared to the dualcell Kranz type C
4
species in adaptation to temperature.
Rather, the efficiency of CO
2
uptake and water use depends on the biochemical type (C
4
, versus intermediate, versus C
3
).
P0226 – Poster
Invasive and native Balsaminaceae species of Baltic region: genetic diversity and factors related to it
Kupcinskiene, E
Zukauskiene, J 1
1 , Jankauskaite, R
, Paulauskas, A 1
1 , Zybartaite, L 1 ,
1 Vytautas Magnus University, Dept of Biology, Kaunas,
Lithuania
Balsaminaceae is mainly distributed in the Old World tropics and subtropics, while the temperate regions in
Europe are inhabited by only a few Impatiens species.
Recent climate change, growing fragmentation of habitats, acidification, nitrogen input or inapproriate management practices including forest logging, alteration of water regimes causes nonindigenous species incorporation into local communities and displacement of indigenous species of similar biology and habitat requirements. Geographical trends in genetics of
Impatiens species are mainly analyzed for Western and
Southern regions of Europe. Nowadays increasing hemeroby of the environment in Baltic countries causes significant changes in distribution of many species including Balsaminaceae representatives. In Baltic
States, data available mainly concerns frequency of
430 invasive species distribution. Impatiens noli-tangere is documented in all districts. Impatiens parviflora is defined as naturalized species of active distribution and is rather frequent in the region: for the first it was mentioned in 1852, and later on in 1920. Impatiens glandulifera runs wild in most part of the Baltic area.
The objectives of this study were: (1) to evaluate genetic affinities of populations of Baltic Impatiens species from different parts of the Baltic region for evidence of local differentiation; (2) to survey local climatic and edaphic data for possible correlates associated with population biology. For the survey of adaptations plant material from 21 populations of Impatiens parviflora , 20 populations from I. glandulifera and 11 populations from
Impatiens noli-tangere was used. The latitudinal gradient covered range of examined species from North (56° 27') to South (53° 54'), a longitudinal gradient extended from
West (20° 56') to East (26° 51') of Baltic region.
Populations that were at least 20 km apart were studied.
Habitat types included shaded mires, forest path sides, riverbanks, trench edges, street sides, waste ground and abandoned gardens. Samples of the plants were taken in the period of 20 July - 8 August. For DNA extractions, at each site, leaves were taken from the tops of 20-22 plants. From the leaves DNA was extracted using a genomic DNA Purification Kit (#KO512, Fermentas,
Lithuania). For PGR reactions Taq polymerase was used
(Fermentas, Lithuania). Among 30 RAPD primers tested the most usefull ones were as follows: OPD-20, OPQ-11,
269, 340, 250, 222, 474, 516, OPB-7, OPA-20
(Biomers.net GmbH, Germany). RAPD information is discussed in relation with ISSR and chloroplast atpBrbcL spacer sequence data. UPGMA dendrograms revealed differentiations between the populations of examined species. Different sets of markers revealed estimates of genetic diversity related to geography of populations. Habitat peculiarities including climatic, edaphic and anthropogenic factors might be important source of genetic variation.
P0227 – ePoster
Phytosociology in rock field of the Iron Quadrangle -
Minas Gerais, Brazil
Pereira, A 1 , Miranda, E 1 , Schaefer, C 1 , Neri, AV 1
1 Universidade Federal de Viçosa, Brasil
Introduction: the Iron Quadrangle is located in a transition area between two Brazilian biomes: Cerrado and Atlantic Forest. By coincidence of areas of ferruginous rock field with iron formations of high commercial value, mineral exploration opportunities without compromising these ecosystems become necessary. The vegetation associated with hematite outcrops in Minas Gerais was named Ferruginous Field
(Rizzini, 1979) and is among the most threatened ecosystems in the state (Jacobi et al., 2008), highlighting the need for its protection. Objectives: This study aimed to characterize the main flora of a piece of ferruginous rock field of the Iron Quadrangle in Minas Gerais, with its horizontal and vertical structures of the tree component. Methodology: The area chosen for this study
(Canga Plateau - Mariana - MG) was analyzed in the degree of conservation and access through geological data, Landsat satellite images and field visits. Where

there is an accumulation of nutrients, organic matter and water, there is an increase in size of vegetation, forming small forestry fields, where 10 plots of 10x10m in different areas were randomly allocated. Individuals with a diameter of 1,30m above the ground less than 3cm circumference were measured, identified and total height estimated. To study the horizontal structure, we used the phytosociological parameters of density, dominance and frequency as well as the importance value. Results: We sampled 705 individuals in 67 species belonging to 45 genera of 26 families. The Shannon index was 3,24 and the evenness index 0,77, indicating good distribution, without significant dominance of any kind. The main families in the phytosociological survey in terms of richness of species were Myrtaceae, with 15 species sampled, followed by Fabaceae (7 species), Lauraceae (5 species), Melastomataceae (4 species), Rubiaceae and
Sapindaceae (3 species each). Regarding the number of sampled individuals, the family Myrtaceae once again stood out, with 201 individuals, representing 31,11% of the total. Next comes Peraceae family, which although it had only one species stood out from the many individuals
(112), representing 17,34% of the total. Fabaceae also stood out with 95 individuals, representing 14,71% of the total. Rubiaceae had 39 individuals, representing 6,04% and Annonaceae with 30 individuals sampled, representing 4,64% of the total. Conclusion: The study area is shown floristically diverse, with great richness of species, some being considered at some risk of extinction. Shannon and Pielou scores calculated were within the quoted literature for other areas of the same formation. However, diversity was higher than in other related areas as in Carajás, in Pará State, confirming the importance of the conservation of this type of vegetation, especially in areas of economic interest.
P0230 – ePoster
It gets hot down at the pig farm, how
Typhonium eliosurum
attracts insect visitors
Barnes, J 1 , Grant, NM 1 , Miller, RE 2 , Robinson, SA 1
1 University of Wollongong, Australia; 2 Monash
University, Australia
Thermogenesis, the phenomenon by which plants produce heat in their reproductive organs, occurs across a broad range of families from ancient Gymnosperms such as the Cycadaceae (1) to Angiosperms from the two major classes; eudicots (e.g. Nelumbonaceae; (2, 3)) and monocots (e.g. Araceae). The Araceae family contains more species of thermogenic plants than any other family
(4), and has thus attracted much attention from researchers aiming to understand heating mechanisms (5-
9), or to characterise the ecological significance of thermogenesis in plant-pollinator interactions (10, 11).
We examined the pollination biology of the Australian native Araceae Typhonium eliosurum (F.Muell. ex
Benth.) to determine whether it is thermogenic, and to identify potential insect pollinators. The pollination biology of T. eliosurum was compared to that of two cooccurring Araceae, the naturalized, Colocasia esculenta
(L.) Schott and the native Alocasia brisbanensis
(F.M.Bailey). Thermogenic profiling, floral development sequences and insect identifications were conducted on these populations located within the Wollongong Botanic
Gardens. Significant heating occurred in T. eliosurum during the two days and nights prior to pollen release with spadix temperatures increasing up to 3 ºC above air temperatures. Maximum heating in T. eliosurum occurred the day before pollen release and was associated with pungent odours resembling pig manure. The single heating period of T. eliosurum contrasted to that of C. esculenta which displayed two maximum heating events, with associated with sweet, fruity odours, during two consecutive mornings before pollen dehiscence. Insects most commonly found in the floral chambers of T. eliosurum were Diptera , in particular the family
Sphaeroceridae. Insects most commonly attracted to A. brisbanensis during flowering were from two Coleoptera families, Nitidulidae and Staphylinidae. No insect pollinators were found within C. esculenta plants. This study is the first to show that T. eliosurum is thermogenic and provides an impetus for future research into the pollination biology of Australian native Araceae species.
Further research is important to gain a stronger understanding of the ecological significance of thermogenesis and insect visitors to the overall reproductive success of these fascinating plants. (1)
Tang, M et al. (1987); (2) Miyake, K (1898); (3) Watling,
JR et al. (2006); (4) Meeuse, BJ (1966); (5) Ito, K et al.
(2003); (6) Onda, Y et al. (2008).; (7) Wagner, AM et al.
(2008); (8) Grant, NM et al. (2008); (9) Miller, R et al.
(2011); (10) Gibernau, M, Barabe, D (2002); (11)
Gottsberger, G (1999).
P0231 – Poster
Plasma membrane Ca
2
+ transporters mediate virusinduced acquired resistance to oxidative stress
Shabala, S 1 , Bækgaard, L 2
Babourina, O 4
, Shabala, L
, Palmgren, MG
Nemchinov, LG 6
2
3 , Fuglsang, A
, Cuin, TA 5
2 ,
, Rengel, Z 4 ,
1 School of Agricultural Science, University of Tasmania,
Australia; 2 Dept of Plant Biology and Biotechnology,
University of Copenhagen, Denmark; 3 Menzies Research
Institute, University of Tasmania, Australia; 4 School of
5
Earth and Environment, University of Western Australia;
CNRS-INRA, Montpellier, France; 6 USDA/ARS,
Molecular Plant Pathology Laboratory, Beltsville, USA
Cross-tolerance is the synergistic co-activation of nonspecific stress-responsive pathways, referring to a situation when an organism’s exposure to one stress increases its tolerance to another. This paper reports the phenomenon of acquired cross-tolerance to oxidative stress in plants and investigates the activity of specific
Ca
2
+ transport systems mediating this phenomenon.
Nicotiana benthamiana plants were infected with Potato virus X (PVX) and exposed to oxidative (either UV-C or
H
2
O
2
) stress. Plant adaptive responses were assessed by the combined application of a range of electrophysiological (non-invasive microelectrode ion flux measurements), biochemical (Ca
2
+- and H+-
ATPase activity), imaging (FLIM measurements of changes in intracellular Ca
2
+ concentrations), pharmacological and cytological (TEM) techniques. Our major findings are as follows: (1) PVX-infected plants show better tolerance to UV stress, showing less structural damage to chloroplasts (revealed by the TEM technique) and less functional damage to PSII (judged by
431

chlorophyll fluorescence Fv/Fm values); (2) PVXinfected plants show better tolerance to H
Acute 5 mM H
2
O
2 net Ca net Ca
2
2
2
O
2
treatment.
treatment has triggered a significant
+ uptake into tobacco mesophyll cells, with peak
+ influx being two-fold higher in mock controls compared with PVX-inoculated segments. At the same time, H
2
O
2 stress also caused much higher K+ efflux from mock controls compared with PVX-inoculated samples. (3) Cytosolic Ca
2
+ levels are lower in PVX inoculated plants under oxidative stress conditions
(estimated by conventional confocal and fluorescence lifetime imaging (FLIM) measurements). (4) Active plasma membrane Ca + transporters mediate plant
2 adaptive responses to oxidative stress. Twenty minutes of
UV-C treatment caused a dramatic increase in the magnitude of net Ca
2
+ uptake when measured immediately after UV stress. Two hours later however, a significant net Ca
2
Ca
2
+ efflux was measured. The kinetics of
+ efflux was strikingly different between control- and
PVX-inoculated leaves, with much rapid ‘switch' to Ca
2
+ pumping in inoculated plants. (5) Plasma membrane
Ca
Ca
2
2
+/H+ exchangers but not Ca
2
+-ATPases mediate net
+ efflux under oxidative stress conditions. No noticeable difference was observed in expression levels of plasma membrane Ca
2
+-ATPases or in their potency to bind CaM between PVX-inoculated and control plants; also, two specific inhibitors for P2B type Ca
2
+-ATPases, erythrosine B and eosin yellow (EY), showed no impact on the patterns of stress-induced Ca
2
+ fluxes. (6) Plasma membrane H+-ATPase-catalysed proton pumping increase in response to UV light and viral infection.
Taken together, our results suggest that more active Ca
2
+ removal from the cytosol in PVX-inoculated plants is essential to plant adaptation to oxidative stress conditions, and that this process is most likely mediated by the plasma membrane Ca
2
+/H+ exchanger. Overall, this study highlights the crucial role of Ca
2
+ efflux systems in acquired resistance to oxidative stress in plants.
P0232 – ePoster
Diversity on rock outcrop in the Parque Estadual Da
Serra Do Brigadeiro, southeast Brazil
V. Neri, A 1 , V. Tinti, B 1
E. Valente, G 1
, M. Moura, V
1 Universidade Federal de Viçosa, Brazil
1 , de Souza, PP 1 ,
Mountain areas are recognized as centers of diversity and endemism. Because of their isolation they are also known as soil island. The landscape on this high altitude complex is often a mosaic of scrubs, rock outcrops, grassland and small patches of forests where the soil is deeper. The objective of this work was to study the variation of flora, structure and diversity in high altitude complex in the Parque Estadual da Serra do Brigadeiro
(PESB). The PESB belongs to the Mantiqueira range between the coordinates 40º20’ and 42º 40’S and 20º 21’ and 21º00’ W, in Zona da Mata in the state of Minas
Gerais. The area is located at an altitude of 1,000 m and surrounded by Atlantic Forest. In the studied area,
Lajinha da Sede, we sampled two physiognomies and three environments: the rock outcrop, the scrub, and the vegetation under the scrub. Studying different physiognomies we needed to use different methods. We
432 used the Braun Blanquet's cover value to study the first and third environment. To study the scrub we used the quadrat method. In the rock outcrop we studied 15 plots
(1x1 m) where all individuals were sampled. In the superior scrub layer, we used 10 quadrats of 5x5 m where all individuals with stem diameter greater than 5 cm at soil height were sampled. In the inferior layer, we used smaller plots (1x1 m) inside of the biggest quadrats.
In this case, all the plants were sampled using the Braun
Blanquet, except those with stem diameter greater than 5 cm at soil height. The depth of substrate was measured to characterize the areas. In total 71 species were found: 31 on the rock outcrop, 25 in the superior scrub layer and 37 in the inferior layer. The main families were: Asteraceae,
Fabaceae, Melastomataceae, Poaceae and Rubiaceae.
Despite presenting the highest richness, the inferior scrub layer had the lowest H’ (Shannon diversity index) and J’
( Pielou’s equability) (2,26 and 0,63). The rock outcrop and the superior scrub layer presented similar values of
H’ (2,65 and 2,48) and J’ (0,79 and 0,76). However, the
Hutcheson T-test shows that the Shannon diversity indices are similar in all areas. This T-test can lead us to believe that three environments are very similar, however a PCA analysis shows that the areas have different floristic composition. In the PCA analysis, the quadrats formed three different groups, each representing one of the environment. The depth varied between the areas: in the rock outcrop the deepest was 5 cm, while in the scrub the depth varied from 35 cm to more than 85 cm. This variable is one of the most important factors for colonization of vascular plants. The soil depth can be considered one of the main factors responsible for the difference in floristic composition between the studied areas and the diversity in this landscape.
P0233 – Poster
1
Spatiotemporal variation in an Australian arid zone flower-visitor network
Wardle, G 1 , Popic, T 1 , Davila, Y
University of Sydney, Australia
1
Mutualistic and indirect interactions are central to structuring ecological communities and maintaining biodiversity. However, the degree of sharing of floral visitors among co-flowering plants is not known for many native communities. In arid Australia, flowering is influenced largely by rain events, resulting in times of mass co-flowering interspersed by times of little flowering. Our data reveals that seasonal variation in rainfall has led to temporal and spatial differences in the composition of the flowering plants in arid zone grasslands of the Simpson Desert. The network of interactions among plants and their floral visitors in these arid zone grasslands was sampled to determine how floral phenology might drive the spatial and temporal scale of variation and overall structure of this community. We observed 1273 flower-visitor interactions (464 were unique) between 61 plant and 159 insect species. The structure of the network was highly nested (0.94 on a scale where 1.0 equates to perfect nesting) indicating specialist species interact with more generalist species. The connectivity distribution was consistent with many species with few interactions and few species with many interactions. Overall the network

exhibited small-world properties of short distances between species and was highly clustered, a pattern consistent with other pollination networks from a recent global review. Only native plants and animals were encountered during our sampling, which provides baseline data on the native interactions influencing arid zone grasslands. The unpredictable rainfall peaks and fires that characterise the arid zone are likely to become more extreme under current models of climate change, which could lead to species range shifts or losses and affect the structure of the network. Continued monitoring of this flower-visitor network will reveal the extent of spatial and temporal variation and whether invasive species, such as the European honey bee, become important components of the plant-pollinator system.
P0234 – Poster
Ground-truthing floral visitor networks – validation is critical to quantifying connectance
Wardle, G 1
S 2
, Gross, C 2 , Hoebee, S 3 , Popic, T 1 , Chivers,
1 University of Sydney, Australia;
England, Armidale, Australia; 3
2 University of New
La Trobe University,
Melbourne, Australia
Despite the rapid progress world wide in producing flower visitor networks, decisions on which species to include are problematic for at least three reasons. Firstly, although species observed visiting flowers are potentially acting as pollinators, many will be ineffective at performing this task and others will often remove floral rewards without contributing to plant reproduction.
These non-mutualistic and indirect interactions are important to the broader ecological function of communities, but without further clarification their unqualified inclusion is problematic for comparative studies that assume that the published visitor networks represent mutualistic pollinator interactions. To address this, we outline a series of validation tests and experiments that a species must fulfill before being included in a pollinator network. The tests are based on the match of floral morphology to visitor size, shape and behaviour, as this will inform the potential for pollen transport. The experiments would further establish that a pollen vector is required, confirm pollen transport and deposition on conspecific stigmas and quantify the strength of the interaction. The key test, however, is to observe whether the species contributing to each link cooccur in time and space – to ensure that forbidden links are not included in the network. Secondly, the current practice of pooling data to report annual networks, or over spatial scales that do not realistically reflect potential function, will inflate the network dimensions and directly bias the connectance, nestedness and the many indices used to analyse networks. Similarly, the exclusion of singleton observations will also influence the structural properties of networks. These decisions taken during the analysis of networks are exacerbated by the problems outlined in the next and final point. Diverse field methods, varying levels of sampling effort, and partitions of diurnal and nocturnal samples will directly influence the number and identity of visits and flowers and flow onto network size and composition. It is acknowledged that this will be true to some extent for all compilations of published work but it is particularly important for comparisons across diverse networks such as mutualistic versus trophic networks, as the empirical approaches will be even more divergent. To illustrate the importance of these issues we use two case studies from our own work on the vegetation of granite outcrops and in arid spinifex-dominated grasslands. The proliferation of networks has sparked wide interest in synthesis and comparisons, and therefore it is timely to encourage field researchers to incorporate explicit ground-truthing and validation of floral visitors as pollinators within these networks. Robust pollinator, pollen-transport, floralvisitor and other networks can then be used to understand the assembly and maintenance of ecological networks, contribute to benchmarks for ecological restoration of degraded areas and to fundamental ideas about how the stability and resilience of the networks will respond to environmental changes.
P0235 – Poster
Alternate stable state theory in kauri forest: kauri as an ecosystem engineer?
Wyse, S 1 , Burns, B 1 , Wright, S 1
1 University of Auckland, New Zealand
New Zealand Kauri ( Agathis australis ) is the largest and longest lived tree species in New Zealand forests, dominating its ecosystems and known to exert a substantial influence on soil properties and nutrient cycling. Although considerable research has investigated the effects of kauri on its soils, less is known about the potential importance of the species in shaping habitats, or its influence on its associated plant communities. These communities are often highly distinctive since species that may be rare in adjacent forest types are common in kauri dominated forest and vice versa, causing marked changes in species composition between sites where kauri is present or absent. The alternative stable state theory proposes the existence of ecosystems in which two or more 'states' can occur for a given set of environmental conditions, with various feedback processes acting to maintain the system in one or another state. The present research aims to determine whether the characteristics of the vegetation in kauri and adjacent forest ecosystems can be explained by this theory with kauri creating its ecosystem state through feedback mechanisms which enhance its persistence. The research will allow assessment of the ecological importance of kauri within its communities, thus determining how potential risk factors that threaten kauri may also threaten the wider plant community. Preliminary results indicate that kauri may significantly influence its neighbouring plants; controlling the suites of species which coexist with it and creating habitat for a range of species which otherwise may not occur in the region.
P0236 – ePoster
Breeding system and its consequence on fruit set of a rare sand dune shrub
Eremosparton songoricum
(Fabaceae: Papilionoideae): implication for conservation
Zhang, DY 1,2 , Shi, X 3 , Wang, J 1 , Liu, H 2
433

1 Key Laboratory of Biogeography and Bioresources In
Arid Land, Xinjiang Institute of Ecology and Geography,
Chinese Academy of Sciences, China; 2 Turpan
Eremophytes Botanical Garden, Chinese Academy of
Sciences,China; 3 Forestry College, College of
Agriculture In Shihezi University, China
The breeding system and its consequence on fruit set of
Eremosparton songoricum (Litv.) Vass., a rare shrubby legume occurring in migratory or semi-fixed sand dunes of Central Asian deserts, were examined by manipulative experiments and observational studies in a natural populations during 2007~2009. Results showed that E. songoricum exhibits a mixed mating system. It is selfcompatible, but depends strictly on pollinators to set fruits. Only two effective pollinators were detected and they triggered the specialized pollination mechanism (a
'brush type' and 'tripping mechanism'). Because
74.5±1.3% of visiting activity happened within or between inflorescence and 24.3±1.4% occurred between ramets, geiotonogamy becomes predominant in natural populations. As a result, inbreeding depression caused by geitonogamous selfing inevitably happened under nature conditions, showing 2.36 times less fruit set than that of hand cross-pollination. The results give light on the rarity mechanism of the speices in relation to breeding system, and help to develop suitable conservation strategies under sever desert environment.
P0237 – ePoster
Evaluation of salt tolerance parameters including growth and physiology in nine table grape (
Vitis vinifera
L.) genotypes
Abbaspour, N 1 , Mohammadkhani, N
, Fozuni, M 1
1
Dolati baneh, H 2
Urmia University, Urmia, Iran;
Research Center, Urmia, Iran
2
1 , Heidari, R 1 ,
Urmia Agricultural
The effect of NaCl salinity on growth, ion content and compatible solutes changes in nine table grape ( Vitis vinifera L.) genotypes (Laal e Bidaneh, Gharashahani,
Sachagh, Shahrudi, Laal e Sefid, Khalili, Chawga,
Ghezeluzum and Gharagandomeh) were studied. These genotypes have been grown in the regions close to Urmia salt lake (West Azarbaijan, Iran) for several decades.
Hydroponicaly grown own rooted cuttings of the vines were treated with 0, 25, 50 and 100 mM NaCl for two weeks. Shoot and root growth and dry weights were significantly (p<0.05) reduced, although at low salinity
(25 mM) growth was stimulated in some cultivars.
Among the genotypes, Ghezeluzum showed a higher growth inhibition and the shoot and root dry weights decline. Chloride and sodium were highly accumulated in
434 the various parts of the vines (root, stem, petiole and lamina) with increasing external NaCl concentration.
Lamina Cl content was generally higher than 1.5 % og
D.W at 100 mM NaCl. However, Gharashahani showed a higher ability to restrict Cl and Na + transport to lamina
(1.08 % of D.W and 2.56 % of D.W respectively) in comparison to the others, particularly to Ghezeluzum
(4.1 % of D.W and 4.5 % of D.W respectively) that showed intensive leaf burn symptom. There were interactions between NO3 and Cl and also between K + and Na + in the various organs determined as a negative correlation (p<0.01). It was shown that root and shoot K +
/Na + selectivity in Chawga was significantly higher than that of the other genotypes. Lamina soluble sugars, proline and glycine betaine contents increased when external NaCl was increased. Correlation analysis
(p<0.01) for each genotype indicated that there was a higher positive correlation (+0.99) between Cl and Na + contents with glycine betaine, proline and soluble sugars accumulation in lamina. In comparison to the other genotypes, Gharashahani accumulated high amounts of glycine betaine in leaf blade particularly at 50 and 100 mM NaCl. As far as low Na + and in particular Cl transport to the shoot as well as high compatible solutes and soluble sugars accumulation are concerned, the results showed that Gharashahani and Ghezeluzum had respectively a higher and a lower capacity to tolerate salt stress when compared to the other genotypes. However all the genotypes studied here seems to be relatively sensitive when exposed to high external salinity.
P0238 – ePoster
Wheat genotypic variation and protein markers in
1 relation with in vitro selection for drought tolerance
Abdel-Hady, MS 1 , Hoda, MHE 1
Botany Dept, National Research Center, Dokki, Cairo,
Egypt
Embryogenic calli isolated from immature embryos of five durum wheat genotypes included one local cultivar
(Benysweif-2), as well as four introduced lines (MSWD-
2, MSWD-9, MSWD-19 and NRCWD-16) were subjected in vitro selection for drought tolerance. The effect of polyethylene glycol (PEG) on the embryogenic cultures and plant regeneration had been investigated.
The results indicated that the relative growth weight of callus was highly significant influenced by differences in durum wheat genotypes and PEG concentrations. The relative growth of callus decreased rapidly as the concentrations of PEG increased. At the concentration of
5% PEG, all cultures of the five genotypes studied could tolerate that level of drought. At the concentration of
10% PEG, MSWD-2 and MSWD-9 gave the highest percentage on callus growth rate, while Benysweif-2 the lowest one. At the highest concentration of 20% PEG, plant regeneration percentage has been succeeded in the genotypes (MSWD-2, MSWD-9 and MSWD-19) with minimal reduction, while (NRCWD-16 and Benysweif-
2) scored highly reduction at that concentration. The plant regeneration is a good parameter to evaluate the effect of drought. The analysis of water soluble, nonsoluble and alcoholic soluble (gliadin) proteins by SDS
PAGE was detected. Some newly protein markers for drought tolerance were induced under different PEG

concentrations, such as water soluble protein bands in the tolerant genotypes MSWD-2 and MSWD-19, respectively. Other water soluble proteins were expressed only at the higher drought stresses in MSWD-2 and
MSWD-9. Moreover, the gliadin proteins were induced in MSWD-9 and MSWD-19, respectively. While three protein bands were induced under the higher concentrations of PEG 10 and 20% in MSWD-2. Mean while, some protein bands disappeared under the highest drought stress in NRCWD-16 and Benysweif-2. The newly induced protein bands could be used as markers for indirect selection to the drought tolerance, and this could support the development of drought tolerant genotypes of durum wheat.
P0239 – ePoster
Ion beam mutagenesis : new innovative technology for mutation breeding
Abe, T 1 , Kazama, Y 1
Y 1
, Hirano, T
1 RIKEN Nishina Center, Japan
1 , Morita, R 1 , Hayashi,
Induced mutations are highly effective techniques to enhance genetic variation and have played a great role in increasing new cultivars among ornamental plants and crops. We found that the ion beams are highly effective in inducing mutations using developing seed embryos at a particular stage after fertilization without damage to other plant tissues. Many types of mutant including albino, sectorial chimera, periclinal chimera, herbicideand salt-tolerant phenotypes in M1 plants in tobacco. The
RIKEN Nishina Center, RI Beam Factory (RIBF) is the one of the biggest facilities to accelerate heavy ions in all over the world since 1986. Heavy-ion beam consists of particles of vary widely in mass, from helium to uranium and beyond. There are two significant differences between photon (
γ
-ray, X-ray) and ion; ion has both mass and charge, by contrast photon has neither one. As the result, they provide different linear energy transfer
(LET); 0.2 to 2 keV/
µ m for
γ ray and X-ray, by contrast large and vary widely for ion beams, e.g. 23 keV/
µ m for
C ion and 640 keV/
µ m for Fe ion. High-LET radiation causes more localized, dense ionization within cells than low-LET radiation. Then ion beam irradiation is expected to produce double-stranded DNA breaks. It is still uncertain whether the repair systems are inactivated, or merely ion-beam lesions are less repairable. Mutations induced by ion beam irradiation at the molecular level have been most extensively studied in mammalian cells.
The frequency of deletion is higher for ion beam irradiation than for
γ
-rays. We found that the ion beam is an excellent tool for mutation breeding. There are the advantage of ion beam mutagenesis, low dose with high survival rates, high mutations rates and a wide variation.
The irradiation treatment given to the various plant materials is short, only seconds or a few minutes, but is enough to induce mutation. A new variety can be obtained by selecting a mutant with a modification to the target characteristic while retaining the existing valuable ones. The time span for breeding can be shortened significantly to two to three yeas. We already put 19 new cultivars on the market in Japan, USA, Canada and EU since 2002. An international heavy-ion plant research consortium has been organized with 135 national user groups and 15 international institutes.
P0240 – ePoster
Ribosomal FISH mapping reveals hybridity in phytoestrogen producing
Curcuma
species from
Thailand
Anamthawat-Jónsson, K 1 , Sunthornchainaksaeng, P 2
1 Faculty of Life and Environmental Sciences, University of Iceland, Reykjavik, Iceland; 2 Faculty of Science,
Mahidol University, Bangkok, Thailand
Species in the genus Curcuma (Zingiberaceae) that are cultivated widely in Thailand for their phytoestrogenproducing rhizomes are called wan-chak-motluk.
Rhizomes of this plant, which are ovoid to ovatespheroidal in shape and about 8–15 cm in diameter, have long been used in Thai traditional medicine for treatment of uterine abnormalities and ovarian hormone deficit.
Products from cultivars of wan-chak-motluk are extensively used for this purpose, especially those belonging to the species C. comosa . Wan-chak-motluk in cultivation is extremely variable, both in rhizome morphology and overall plant appearance. This has often resulted in wrong identification of cultivars – the feature which is used by local growers to indicate medicinal properties. Botanically speaking wan-chak-motluk belongs to three Curcuma species: C. comosa , C. elata and C. latifolia , but the latter two species have not been systematically explored for their medicinal properties.
The aim of the present study was to compare wan-chakmotluk cultivars from different species and cytotypes using a molecular cytogenetic approach. Five cultivars belonging to C. comosa (cultivars with 2n = 42 and 63) and C. elata (2n = 63) were examined using the molecular cytogenetic method of fluorescence in situ hybridisation (FISH) in order to identify genetic relationships among these cultivars based on chromosomal maps of the 18S-25S ribosomal loci. The results revealed hybrid features in this Curcuma species group and a significant similarity among wan-chakmotluk cultivars. The main features included (1) the presence of the single largest ribosomal site, assigned the
Cc1 marker site, in the somatic 2n complement of all cultivars, and (2) the odd numbers of ribosomal sites in the complements, most often in sets of three. We therefore propose that the cultivar with 2n = 42 ( C. comosa ) is a homoploid hybrid species comprised of two different ancestral genomes and has a diploid status with the (secondary) basic chromosome number x = 21. On the other hand, the cultivars with 2n = 63 ( C. comosa and
C. elata ) are most probably triploids arising within the 2n
= 42 diploid species/cultivars via a meiotic modification, rather than from hybridisation between diploid and tetraploid plants. As homoploid hybrid species or triploid cultivars, wan-chak-motluk should have no difficulty in becoming established in a relatively short time. Wanchak-motluk is cultivated on a large scale via vegetative
(rhizome/bulbil) propagation. Growers might have also selected and preferred certain cultivars, either based on desirable agronomic features or medicinal properties.
The knowledge about genetic and genomic relationships among wan-chak-motluk cultivars will be important in
435

the research projects that aim to explore and promote new plant materials for cultivation.
P0241 – ePoster
Genetic improvement of
Centella asiatica
(L.) urban for high yield of asiaticoside and madecassosides through ploidy breeding
Boddu, JA 1
1 Jijamata College of Science and Arts, Bhende village,
India
Centella asiatica (L.) Urban (syn. Hydrocotyle asiatica
L., family Apiaceae; 2N=22), commonly known as
Pennywort, Indian Pennywort, (Indian names
Mandukaparni, Brahmi and Gotu Kola), is a valuable medicinal plant, highly valued for its glycosides
(triterpenoids), asiaticoside and madecassoside. As the induced ployploids were known to produce increased contents of therapleutically active compounds, attempts were made in the investigation to induce autrotetraploidy in Centella asiatica , employing colchicine, with an objective of obtaining increased production of secondary metabolites, including asiaticoside and madecassoside.
Young developing shoots of Centella asiatica were treated with different concentrations of colchicine for varying time intervals. It was found that treatment with
0.4 % colchicine for 6 hours at room temperature was very effective in inducing autotetraploidy in Centella .
The induced autotetraploids were bigger in size and possessed large leaves with thick petioles. They showed four-fold increase in leaf area, large stomata, 2.5 fold increase in fresh weight, 2 fold increase in dry weight and doubled chromosome number as compared to control diploid plants. The autotetraploid plants were allowed to reproduce vegetatively for at least 15 generations. Such plants, along with their controls were, analyzed on High
Performance Thin Layer Chromatography (HPTLC) for asiaticoside and madecassoside contents. TLC densitometric analysis revealed that the induced autotetraploids produced double the amount of asiaticoside and one and half time more the content of madecassoside as compared to control diploid plants.
P0244 – ePoster
Quercetin accumulation predicts drought resistance in a novel white clover hybrid
Ballizany, W 1 , Hofmann, R 1 , Barrett, B 2 , Jahufer, Z 2
1 Faculty of Agriculture and Life Sciences, Lincoln
2 University, Lincoln, New Zealand; AgResearch
Grasslands Research Centre, Palmerston North, New
Zealand
White clover ( Trifolium repens L.) is an important pasture legume crop in New Zealand. Meat and milk producing livestock performance improves with increasing clover content in swards, by providing valuable proteins. Furthermore, white clover Rhyzobium root nodules fix nitrogen from the air into the soil, lessening the need for fertilisers. Increasing climate change is causing the Eastern and central areas in New
Zealand to become drier, combined with rising levels of
436 damaging UV radiation as a result of diminishing ozone layers. Consequently, new white clover cultivars need to be more resistant to abiotic stresses than ever before.
Molecular plant breeding methods can increase breeding efficiency and identify genes controlling complex traits based on genotype rather than phenotype. Previous research showed that levels of the flavonol quercetin (Q) were positively associated with UV stress resistance in populations of white clover, and were inversely linked to biomass production. Q has been implicated as an antioxidant and energy dissipating compound in plants exposed to stress, and could be a useful indicator for resistance to other stress factors. This study used a novel white clover hybrid of the productive cultivar Kopu II and the stress-resistant population Tienshan to examine whether these relationships also hold under drought on a genotype level for germ-plasm development in future breeding trials. The results showed for the first time that drought resistance in a new white clover hybrid can be linked to drought-induced increases in Q levels, and genotypes that are high in both the traits Q and biomass have been determined. Subsequent DNA-marker analysis studies will be utilised to identify marker-trait associations and quantitative trait loci (QTL) for improved marker-assisted selection (MAS) methods in drought resistant white clover to combine high Q response with high herbage yield.
P0248 – ePoster
Effect of vermicompost on dark green leafy vegetable
(
Spinacia oleracea
L.)
Biradar, SR 1
1 Shri Chhatrapati Shivaji College,Omerga Dist., India
Solid wastes are produced in enormous quantities at a rate of ½ to 1 kg per head per day in Indian society. A major portion of the solid waste is street garbage comprising of leaves, grasses, paper and garbage from kitchen and 4 to 5% is from vegetables, fruits and other market wastes. This solid waste is biodegradable and can be used as a raw material for the preparation of vermicompost. (Kalambasa et al., 1993). A dumping of the biodegradable waste arbitrarily in muncipal areas cause serious environmental health problems like plague and dengue. Vermicompost will not only improve the soil fertility and water retention but also reduce the outlay on chemical fertilizer (Vikram Reddy, 1999).
Many earthworm breeders claim that the produced earthworm compost has significant properties.
Vermicompost was prepared by layering method.
Earthworms of the species Eisenia foetida , the red earthworm or surface dweller, were used. (Kalambasa,
1996). Vermicompost was prepared in a 2’x2’x2' wooden box with bottom holes for aeration. The layering was a
7.5 cm bottom layer of coir followed by a 5 cm layer of cow dung manure uniformly spread over it. 500 earthworms were left on the dung manure. Fresh cow dung slurry was added evenly on it followed by semidigested organic matter, evenly distributed forming
2.5 cm. The final layer was 3 kg of organic matter which included kitchen waste, vegetable waste, garden waste and fruit peds. Water was sprinkled on an even layer to keep it moist. After 15 days the layering was done again with the garbage or kitchen waste. Sufficient water was

sprinkled above it and the whole layering was covered with damp gunny bag. The box was kept in shade and was protected from rats, frogs, dogs, hens, etc. The layering was repeated after every 15 days. After filling the box the total contents were kept moist and it was turned into vermicompost within 45 days. The total waste used from a five member family. The fresh yield on vermicompost, along with dry leaves, was less than the control because the dry leaves could not get sufficient time for the decomposition; however, the fresh yield, dry matter yield and total nitrogen in spinach cultivated on only vermicompost show the significant results over control. The re-growth yield of fresh weight after 95 days showed the significant yield over control and the same sequence was followed for dry matter and nitrogen content in the spinach leaves. In the present investigation the vermicompost was used as a nutrient source for spinach in pots and compared with 50% vermicompost and dry leaves in the garden and control with three replicates.
P0249 – ePoster
Heavy metal contaminants in herbal remedies: preliminary investigation of two concoctions in
Guyana
Caesar, JC 1 , Yhan, A 1
1 University of Guyana, South America
Herbal remedies, popularly referred to locally as bush medicine, have been used for centuries to treat different medical conditions in humans and domestic animals with varying degrees of efficacy. Toxic contaminants in herbal medicines have been reported in the global literature, but may not have been reported in Guyana previously. We examined the presence of heavy metals in selected local herbal remedies – 'cold medicine' and the aphrodisiac
'Kapadula tonic.' Samples were purchased from Bourda
Market and Stabroek Market in the city of Georgetown,
Guyana. Heavy metal contents were analyzed at the
Central Research Laboratory of the Guyana Sugar
Corporation (GuySuco) located at La Bonne Intention
(L.B.I.) Estate on East Coast of Demerara, Greater
Georgetown. Acid digests of 5 replicate samples each of the two herbal remedies were analyzed by means of a
UV-VIS spectrophotometer following AOAC standard procedures. The results confirmed the presence of aluminium, cadmium, iron, zinc, and lead at very high concentrations, each exceeding the respective recommended daily allowance (RDA) values several fold. Mean values of heavy metal contents of samples obtained from the two markets were: Al = 82.79 mg L -1
(Bourda) and 117.23 mg L -1
1 (Bourda) and 10.10 mg L
L -1 (Bourda) and 43.85 mg L mg L -1
21.03 mg L -1
(Stabroek); Cd = 9.00 mg L
-1 (Stabroek); Fe = 38.43 mg
-1 (Stabroek); Pb = 153.87
(Bourda) and 193.10 mg L -1
(Bourda) and 24.76 mg L -1
-
(Stabroek); Zn =
(Stabroek). We intend to expand the survey in order to provide policy guidance for risk communication, safety measures, enhanced safety of 'ethnomanufacturing' methods and human well-being.
P0250 – ePoster
Edible wild plants in Calabria (S Italy): a check-list
1
Caruso, G 1
Dept of Environmental and Crop Science, Marche
Polytechnic University, Ancona, Italy
Calabria is the southernmost region of continental Italy.
It forms a narrow peninsula N-S oriented, surrounded by around 740 km of sea coasts (Ionian and Thyrrenian seas). Inland is mainly hilly and mountainous, while plains are quite rare. Calabria is characterized by mountain ranges up to 2000 m a.s.l., limestone in the north and acid rocks in the centre-south. Climate is mainly thermomediterranean on the coasts gradually shifting into mesomediterranean, supramediterranean, supratemperate, orotemperate towards the mountain tops and Ionian coast is less wet than the Thyrrenian. Vascular flora is quite rich (more than 2600 species) because of altitudinal, geological, climatic diversity as well as geological and biological history. 900 species belonging to Calabrian vascular flora are potentially wild food plants but only 140 species (15.6%) are known to be actually used, today or in the past, in this territory. The remaining 760 species (84.4%), despite not used as edibles in Calabria, are well known as food plants in the surrounding areas (Italian territory, Mediterranean Basin) or even rather far from the investigated area (America,
Asia, Africa). The low level of knowledge about a so high potential of food plants in Calabria is mainly due to lack of ethnobotanical investigation, progressive lost of ethnobotanical knowledge due to globalization, crisis of traditional life-styles, people migration from inland towards coastal settlements. On the other hand ethnobotanical knowledge in Calabria should be hopefully improved in order to increase practical wild food plants applications. Efforts should be concentrated on field ethnobotanical research, rediscovering of traditional food habits using wild plants as part of the local cultural identity, spreading of wild food plant knowledge into different social context (education, agriculture, tourism, trade, etc.), experimenting of new potential applications of wild food plants (cultivation, new recipes, nutraceuticals, food-medicine, etc.)
P0252 – ePoster
Development of tissue culture protocol for promising interspecific F1 tri-hybrid of
Eucalyptus
FRI-13 (
E.
1 camaldulensis
x
E. tereticornis
) x
E. grandis
Chauahn, S 1 , Sharma, R 1 , Rathi, N 1 , Arya, S 2
Forest Research Institute, Dehradun, Uttarakhand,
India; 2 Arid Forest Research Institute, Jodhpur,
Rajasthan, India
Tissue culture, an important aspect of biotechnology has a great potential for rapid and large-scale clonal production of plants. Promising interspecific F1 trihybrid of Eucalyptus artificially developed for the first time in India by Forest Research Institute, Dehradun has displayed a very high degree of vigour (positive hetrosis) both in height, diameter and wood quality. It is very widely adaptable and drought resistant and has fast
437

growth rate and excellent stem form. Technique for successful micropropagation of FRI-13 ( E.camaldulensis
x E. tereticornis ) x E.grandis
from nodal segments has been developed. Explant was collected from selected mature F1 tri-hybrid of Eucalyptus FRI-13 and washed with teepol. Surface sterilization with 0.1% HgCl2 for
10–15 minutes after treating with fungicide (Bavistin) and antibiotics (Streptomycin and Chloramphenicol) were very effective to control 80% contamination with good survival rate (70-75%). Axillary buds of F1 trihybrid FRI-13 were inoculated aseptically on defined
MS medium for axillary bud break. Axillary bud break was achieved within three week of culture on MS medium containing cytokinin BAP with 75% success.
These axillary shoots were later isolated and subcultured on MS medium supplemented with varied concentrations of BAP for in vitro shoot multiplication. The in vitro shoots prior to their subculture on rooting medium need to be elongated. For shoot elongation, these shoots were transferred on MS medium without growth regulator.
After four weeks, the shoots were elongated and were transferred on rooting medium. In vitro rooting was achieved 80% on ½ MS medium supplemented with different concentrations of IBA. Clonally propagated hybrid has been planted in different climatic conditions at different agro climatic sites. So far data collected has shown overall superiority of these hybrids even under varied climatic conditions.
P0254 – ePoster
Evidence for prehistoric human contact between
Polynesia and South America: DNA analysis of the
Sweet Potato (
Ipomoea batatas
)
1
Clarke, AC 1
Nakatani, M 4
, Holland, BR 2
, Matthews, PJ 5
, McLenachan, PA 3
, Green, RC 6 , Penny, D 3
,
2
University of Otago, Dunedin, New Zealand;
University of Tasmania, Hobart, Australia;
University, Palmerston North, New Zealand;
Institute of Crop Science, Tsukuba, Japan;
Museum of Ethnology, Osaka, Japan; 6
4
5
3 Massey
National
National
University of
Auckland, New Zealand
The Sweet Potato ( Ipomoea batatas ) was a crop species fundamental to many agricultural systems in pre-
European Polynesia, but the long-standing scientific interest in this species centres on its origins. The sweet potato is thought to have arrived in the Pacific from
South America, and various lines of botanical, archaeological, linguistic and genetic evidence are consistent with this transfer being effected by Polynesian voyagers who collected the sweet potato from South
America between AD 1000 and 1200. The strongest evidence for contact between Polynesians and indigenous
South Americans is the Polynesian word ‘kumara’, which appears to be derived from the Ecuadorian
‘cumar’. Despite growing acceptance of this humanmediated transfer, there remain a number of unresolved questions about the sweet potato in Oceania, including: the point on the South American coast where Polynesians made landfall, the number of Polynesian lineages introduced into the Pacific, the dispersal routes of sweet potato within Polynesia, and the influence of Spanish
(camote) and Portuguese (batata) introductions of sweet potato in the western Pacific. To address these questions
438 we have used the amplified fragment length polymorphism (AFLP) fingerprinting technique to genotype 270 unique accessions of sweet potato from
Asia, Island Melanesia, Polynesia and the Americas.
AFLP data have been used to construct phylogenetic trees, and to improve phylogenetic resolution we have developed a new method to optimise AFLP scoring parameters. A putative kumara lineage, representing a prehistoric, Polynesian-mediated introduction from South
America, was identified. Sweet potato accessions from
Asia to Western Polynesia were found to be genetically diverse, and the relationships between them are complex.
The phylogenetic positions of the New Zealand M ā ori varieties raises questions about their presumed close relationships with other lineages in Eastern Polynesia.
Together, these findings suggest a much more complex picture of sweet potato dispersal in Oceania than is currently recognised. The current sampling appears insufficient to accurately determine the point of
Polynesian contact on the South American coast, and future work is focused on more intensive sampling to address this. Further work is also planned to better integrate the genetic data with existing historical and linguistic data.
P0255 – ePoster
Structural and functional characterization of photosynthesis in
Oryza
lines
Giuliani, R 1
AB 1
, Voznesenskaya, E
, Edwards, GE 1
2 , Koteyeva, N 2 , Cousins,
1 Washington State University,USA; 2 VL Komarov
Botanical Institute of Russian Academy of Sciences
The genus Oryza is composed of 24 species, including the cultivated O. sativa and O. glaberrima , which belong to several genomic groups and are known to have C
3
type photosynthesis. The species exhibit a wide range of phenotypes, including annual versus perennial life cycles, and sun versus shade-adapted species. The aims of this study are 1) to describe multiple leaf structural and functional features of 17 Oryza species, 2) to group the species based on structural, functional, and structuralfunctional clusters of traits, 3) to analyse the functional significance of structural diversity, 4) to evaluate whether structural-functional relationships reflect genomic or habitat (e.g. sun/shade) of Oryza species, and 5) to determine whether these structure-function relationships influence rates of photorespiration. For each species, analyses were conducted on leaves from 30-40 day-old potted plants grown under controlled conditions. Leaf and cell structural traits were determined with light and electron microscopy, including the quantification of mesophyll cell surface area facing the intercellular air spaces (IAS) and its fraction covered by chloroplasts
(expressed per unit leaf surface area), thickness of lamina and of chlorenchyma cell walls, and cell size. Functional photosynthetic characteristics were determined from leaf gas exchange measurements over a range of partial pressure of CO
2
and at 20 and 40% O
2
. The CO
2 compensation points, maximum carboxylation efficiencies, and photosynthetic rates at ambient and saturating levels of CO
2
were determined. Stomatal and mesophyll conductance to CO
2
were analyzed (the latter from combined leaf gas exchange and chlorophyll

fluorescence measurements). Additionally, intrinsic water-use efficiency was calculated from the ratio of leaf net photosynthetic activity and stomatal conductance to water vapour. Data analyses were carried out to first explore the variance in each functional and structural trait across and within Oryza genomic groups. Based on traitto-trait correlation analysis significant structural and functional traits will be extracted to characterize each
Oryza species. Furthermore, multivariate analyses will be performed to cluster species based on selected structural and functional traits. The functional significance of leaf structural heterogeneity and the clusters will be assessed and discussed with respect to Oryza genomic groups.
P0256 – ePoster
HS-SPME-GC-FID based method development for quantification of 2-acetyl-1-pyrroline synthesized by
Bacillus cereus
(ATCC 10702)
Deshmukh, Y 1 , Nadaf, A 1
1 University of Pune, India
2-acetyl-1-pyrroline (2AP) has been identified as a principle basmati aroma compound in basmati and other scented rice varieties. Bacillus cereus (ATCC 10702) has been reported to synthesize 2AP. A simple and reliable methodology for the extraction, detection and quantification of 2-acetyl-1-pyrroline, synthesised by
Bacillus cereus has been developed using headspace solid-phase micro-extraction technique (HS-SPME) coupled with gas chromatography flame ionized detection (GC-FID). The optimization has been carried for the several parameters, viz. medium for bacterial growth, bacterial incubation period, sample weight, adsorption time and temperature, head space equilibration time and different precursors with different concentrations. The optimum conditions for maximum extraction of 2AP were 500 mg of 48 hr grown B. cereus on plate count agar supplemented with 4% ornithine,
80°C adsorption temperature, 10 min of headspace equilibration time and 25 min adsorption time. Following these optimized conditions, 2AP was quantified for B. cereus (ATCC 10702) as 0.21 ppm within 50 min per sample. Thus, this method is rapid solvent free, fast and requires less quantity of sample. The developed method can be used for large scale screening and quantification of 2AP synthesized by different bacterial isolates.
P0257 – ePoster
Effects of different harvesting methods in growth and productivity of sphagnum moss
Diaz, MF 1 , Silva, W 2
1
2
Universidad Andres Bello, Caracas, Venezuela;
Universidad de Chile
Sphagnum secondary peatlands have become major sites for commercial harvesting of Sphagnum moss by local communities in southern Chile. Sphagnum moss species, primarily Sphagnum magellanicum , is used internationally as substrates for horticulture. Chile exports annually 2,250 tons of dry Sphagnum moss derived from the southern temperate forest region.
Because of the growing international market demand, there is an indiscriminate use of the resource, with a number of potentially negative ecological and social consequences, such as disruption of Sphagnum ecosystems, changing water storage capacity of moss wetlands affecting water supply to rural communities, and reducing biodiversity. In Chile moss extraction has proceeded for nearly 10 years without technical protocols to insure regeneration of the resource. Moss extraction is showing symptoms of overexploitation and deterioration of sustaining conditions for moss production. We urgently need to promote a sustainable management protocols to ensure the economic and ecological viability of resource use in the long term. There is no government office that currently regulates Sphagnum moss extraction from private lands. To fill in the sustainability gap due to the current lack of regulation and high demand for this resource, we attempt to improve harvesting techniques to ensure moss regeneration and the sustainability of moss cover in harvested areas. What is the influence of different harvesting methods on the growth rates of moss? Is moss regrowth promoted by re-seeding in harvested sites? Answering these questions can enable us to improve the existing harvesting methods and formulate future management plans for the sustainable use of Sphagnum moss in peatlands. To address these questions, we determined growth rates and biomass production of Sphagnum moss in harvested and nonharvested sites and evaluated the influence of re-seeding on the recovery rate of Sphagnum after harvesting. We installed 28 3m 2 plots to determine growth and biomass production. Each plot with 3 subplots as treatments: control, without harvest (C), with harvest (H), and with harvest and re-seeding (R). Data were analyzed by a randomized block design analysis of variance. Moss growth is very slow compared with other studies in the region. The mean cumulative growth of control plots was
2.33 cm year -1 , higher than harvested plots. There is no differences between harvest and harvest with re-seeding plots. Analyzing biomass production, we found differences between treatments. Control plots recorded the highest biomass production. Our results suggest that it is extremely important to include biomass production and not only height increment or growth when considering the right time to harvest. If only the growth in length is used as a proxy of the decision to harvest or not, overexploitation of the resource will be imminent.
P0259 – ePoster
Distribution, properties and synthesis of (1,3;1,4)-
β
-
D-glucan in
Sorghum bicolor
(L.) Moench
Ermawar, R 1 , Collins, H 1 , Fincher, G 1 , Burton, R 1
1 ARC Centre of Excellence In Plant Cell Walls, School of
Agricuture, Food and Wine, Waite Campus, University of
Adelaide, Australia
The amount, properties, distribution and biosynthesis of
Sorghum bicolor (1,3;1,4)-
β
-D-glucan were examined in this present study. The amount of the (1,3;1,4)-â-Dglucans in five varieties of Sorghum grain and vegetative tissues was examined using a modified Megazyme assay.
In grain, the amount was low while in the vegetative tissues it was found to vary and was significantly influenced by the age of the tissue. Properties of the S. bicolor (1,3;1,4)-â-glucan were indicated by the
439

DP3:DP4 ratios in both the grain and the vegetative tissues where the ratios appeared to be varied and influenced by temporal and spatial factors. Distribution of the (1,3;1,4)-â-D-glucans in S. bicolor was examined using the specific antibody BG-1 and visualised with fluorescence and transmission electron microscopy
(TEM). The fluorescence and TEM micrographs also showed a variable distribution of (1,3;1,4)-â-D-glucans both in grain and vegetative tissues linked to both temporal and spatial factors. Real-time quantitative PCR
(Q-PCR) analyses of the Sorghum cellulose synthase-like
F and H gene families showed that the transcript of the
SbCslF6 gene was dominant in most tissues except in the older leaf where the level of one of the CslH genes was highest. In the developing grain the SbCslF6 transcript was also dominant where it peaks around the soft dough stage of development.
P0263 – ePoster
Predicting climate change impacts on yield and nutritional value of cyanogenic tropical crops
Gleadow, R 1
Blomstedt, C 1
1
Cavagnaro, T 1
, Miller, R 1 , Zacarias, A 2
, Hamill, J 1
, O’Donnell, N
, Evans, J 3 , Burns, A
1
1
School of Biological Sciences Monash University,
Australia; 2 Agricultural Research Institute of
Mozambique (IIAM), South Africa;
Biology, ANU, Canberra, Australia
3 Research School of
,
,
Tropical crops will become increasing important for food production as the planet warms and traditional food growing regions in temperate latitudes become progressively drier. Sorghum and cassava are contenders for meeting this gap. Both are have good potential as biofuels crops. With the dwindling amount of arable land and the increasing cost of fertilisers, research to date has largely focused on increasing yields, but food quality also needs to be considered. These crops are among dozens of crops that produce cyanogenic glucosides that break down to release toxic cyanide gas and can be detrimental to human and animal health. Cases of cyanide poisoning due to consumption of high cyanide varieties of cassava have been associated with epidemics of the permanent paralysing disease konzo, which particularly affects children and women of child-bearingage and sometimes death. Sorghum grain is not cyanogenic, but the leaves can be lethal to grazing animals. The concentration of the cyanogenic glycosides depends on environmental conditions as well as genotype, ontogeny and phenology. We are creating a framework to facilitate predictions for the future nutritional value and productivity of these crops by coupling ecophysiological studies in glasshouses with climate models of southern Africa. Both species become more toxic when subject to water stress and have reduced yields. At elevated CO
2
cassava, a C nitrogen use efficiency. Sorghum , a C
3
plant, becomes more cyanogenic probably as a consequence of improved
4
plant, becomes slightly less toxic when grown at elevated CO
2
, probably due to increases in water use efficiency. Adaptation strategies to avoid cyanide poisoning from cassava include development and implementation of low-cyanide varieties of cassava, improved processing of cassava products and wider testing of those products.
440
Diversification of the diet of cassava-dependent communities is also going to be increasingly important as
S-containing amino acids are required for the detoxification of cyanide. Adaptation strategies for forage Sorghum lie primarily in the development of new varieties that are less toxic, and improved field monitoring so that grazing can be minimised during periods of water stress. This requires an interdisciplinary approach. As cyanogenic crops are introduced to new areas, it is important that this is accompanied by knowledge about how to manage their toxicity.
P0264 – ePoster
Carbon storage character of major bamboo forest and management technology of carbon storage forest type in China
Feng, Y 1
1 ICBR, China
After overall investigation in bamboo forest distribution and resource situation, we divided the bamboo into bamboo culm, bamboo branch, bamboo leaf, culm stump, bamboo rhizome, bamboo root. We also can divide it into plant layer, litter layer and soil layer, or into aboveground and bellow-ground. This investigation covered all the bamboo distribution area, including 16 bamboo forest types belonging to 5 bamboo area. In these bamboo area, biomass, carbon density and carbon pattern were researched in 172 permanent plots or temporary sample plot. Six bamboo including Phyllostachys edulis were done in eco-geographical regions, stand management degree and carbon character.
P0266 – ePoster
Satureja khuzistanica
: a valuable medicinal plant for rural development in Iran
Hadian, J 1 , Najafi, F 1 , Salehnia, A 1 , Ehteshamnia, A 1
1 Shahid Beheshty University, Iran
Satureja khuzistanica , an endemic herb with interesting pharmacological and biological properties, grows wild in south-west of Iran. A domestication program has been conducted to develop improved varieties and cultivation guideline in marginal lands. In first step, morphological and phytochemical variability was studied among and within populations. Among all populations, those of
Abdanan (2.81 %) and Kaver (2.79 %) showed the highest oil yield. Essential oils analyzed by GC-FID and
GC–MS showed that all 69 sampled individuals among populations have the high percentage of carvacrol
(ranging from 89.59 % to 95.41 %) as main component.
Talent populations were sampled and transferred to field where the genotypes were evaluated in two successive years. Several genotypes with high oil and biomass yield were identified and propagated through tissue culture to make clones. Parallel with vegetative propagation and cultivation, selection will be performed to gain homogenous seed variety. Several experiments have been conducted to define nutrient and irrigation requirements, plant density, etc., to develop guideline for agriculture production. Based on our primarily results, S.

khuzistanica is a good plant of choice for cultivation in marginal land and rural development in south-west of
Iran.
P0267 – ePoster
Variability of morphological trait and rosmarinic acid content of
Satureja khuzistanica
Jamzad populations from Iran
Hadian, J 1 , Salehnia, A 2
1 Shahid Beheshty University, Iran; 2 Dept of Research and Development (R&D), Khorraman Pharmaceutical
Co, Khoramabad Industrial City, Iran
Satureja khuzistanica , an endemic herb with concentration of biologically active compounds growsing wild in Iran. Morphological and phytochemical variability was studied among and within populations.
Among populations, peduncle length (cv. of 47.39 %) and leaf surface area (cv. of 47.21 %) were the most variable characteristics. Rosmarinic acid content of methanolic extracts, analyzed by means of HPTLC, showed high level of variability (cv. of 50.0 %) ranging from 1.81 % in populations of Abdanan to 0.59 % in population of Paalam. Our results revealed high level of morphological and phytochemical variability among and within populations. This suggests breeding approach during domestication to gain new promising homogenous cultivars, attractive for industry and agriculture.
P0269 – ePoster
Analysis of population structure and genetic diversity within a global collection of Tall Fescue (
Festuca arundinacea
Schreb.)
Hand, ML 1,2,3 , Cogan, NOI 2 , Forster, JW 3
1 Dept of Primary Industries, Victorian AgriBiosciences
Centre, La Trobe University Research and Development
Park, Bundoora, Australia; 2 Dairy Futures Cooperative
Research Centre, Australia; 3 La Trobe University,
Bundoora, Australia
The Festuca genus contains over 500 species of temperate grasses, that range in ploidy level from diploid
(2n = 2x = 14) to dodecaploid (2n = 12x = 84). The most agriculturally important Festuca species is Tall Fescue
( Festuca arundinacea Schreb.), an outbreeding allohexaploid (2n = 6x = 42) cultivated for pasture worldwide. The majority of cultivars are classified as
Continental types, and have arisen from northern
European ecotypes that display maximum growth over summer periods. All additional cultivars can generally be categorized as either Mediterranean or rhizomatous, which differ from Continental cultivars with respect to geographical origin, growth patterns and morphological features. Comparison of homoeologous nuclear gene sequences from each tall fescue morphotype has revealed that the Continental and Mediterranean types are likely to have arisen independently following hybridization of different progenitor taxa; a finding which supports observations of hybrid sterility. Previous sequencing of chloroplast and internal transcribed spacer region from each tall fescue morphotype and related tall fescue cytotypes has also identified diagnostic nucleotides capable of characterising tall fescue germplasm. This approach has been used to screen a large collection of tall fescue and meadow fescue accessions held and maintained by the United States Department of
Agriculture through the Germplasm Resource
Information Network. Results have identified the presence of each three tall fescue morphotypes as well tall fescue sub-species of varying ploidy levels in both collections. Characterisation of these accessions has enabled biogeographical analysis of each tall fescue morphotype and sub-species and as expected, has revealed a North African distribution of Mediterranean tall fescue and a wider European distribution of
Continental tall fescue from Spain through to western
China. Genetic structure within Continental,
Mediterranean and rhizomatous tall fescue was investigated further through the amplification of 34 SSR markers that had originally been developed from perennial ryegrass sequence. This analysis supports the divergence between Continental and Mediterranean tall fescue and also identifies subpopulations within the
Continental germplasm that reflect morphological differences and geographical distribution. This work has provided an insight into the phylogeographical history of tall fescue whilst comprehensively characterising and dissecting a complex germplasm resource for the benefit of plant breeding.
P0271 – ePoster
Ectopic expression of NAD biosynthetic gene causes developmental alterations in
Arabidopsis
; a precocious ageing in leaves and inflorescences
Hashida, SN 1 , Takahara, K 2 , Kawai-Yamada, M 3 ,
Kitazaki, K 1
Uchimiya, H 4
, Shoji, K 1 , Goto, F 1 , Yoshihara, T 1 ,
1 Central Research Institute of Electric Power Industry,
Tokyo, Japan; 2 Institute of Molecular and Cellular
Biosciences, University of Tokyo, Japan;
Environmental Science and Human Engineering,
Saitama University, Japan;
3 Dept of
4 Institute for Environmental
Science and Technology, Saitama University, Japan
Nicotinamide adenine dinucleotide (NAD(P)(H)) plays indespensable roles in growth regulation and responses to various stressful environments. It is recently reported that overexpression of NADP biosynthetic gene promoted carbon and nitrogen metabolism without significant morphological alternations in Arabidopsis and rice. In this study, we generated transgenic Arabidopsis plants overexpressing NAD biosynthetic genes, NMNAT and
NADS. Ectopic expression of NMNAT and NADS did not show any remarkable changes in vegetative growth.
However, ectopic NADS expression induced early ageing just after bolting. Noticeably, the ageing was detected not only at leaves but also at inflorescences and stems. Consequently, NADS-overexpressing plants frequently failed to set a seed. Transcriptional analysis revealed that gene expressions of senescence accelerated genes were significantly upregulated and the expressions of senescence down-regulated genes were precociously decreased. Interestingly, the expressions of salvage NAD synthesis genes were dramatically increased.
Metabolome analysis demonstrated that the levels of
441

intermediates of NAD biosynthesis (NaMN and NaAD) were dramatically increased after bolting in NADSoverexpressing plants only while the level of NAD was comparable to wild type. Alternatively, NAD derivatives
(nicotinate and nicotinamide) were accumulated in
NADS-overexpression plants. These results suggest that the ectopic NADS expression causes futile NAD turnover and indicate a possible role of alteration of
NAD homeostasis as a signal for the phase transition from vegetative growth to reproductive growth
P0272 – ePoster
Effect of heavy–ion beam irradiation on mutation induction in rice
Hayashi, Y 1 , Morita, R 1 , Tokairin, H 1 , Sato, T 2 , Abe, T 1
1 RIKEN Nishina Center, Wako-shi, Japan; 2 Graduate
School of Life Sciences, Tohoku Univ., Sendai, Japan
Heavy ion beams have high linear energy transfer (LET) and produce more dense and localized ionization along their tracks than other radiation such as X-rays and
γ
-rays for the same dose. Therefore, a heavy ion particle is postulated to cause double strand break of DNA and induce mutation with low dose irradiation. LET of ion beams is an important factor affecting mutagenesis. We examined the effect of LET on mutation induction in rice. The seeds of rice ( Oryza sativa L. cv. Nipponbare ) were soaked for 3days in water at 28 ° C without light. The single-layered seeds were exposed to C or Ne-ion accelerated to 135 MeV/u by RIKEN Ring Cyclotron at
RI-Beam Factory. The dose range and the LET range of the beam were 5 to 40 Gy and 22.5 to 100 keV/
µ m respectively. The LET values were calculated at the surface of the seeds. M1 plants were grown in the paddy field and harvested individually five month after transplanted. Survival rates were surveyed in M1 plants.
The seed fertilities were surveyed with a main panicle from each plant. The mutation rate was estimated by the frequency of M1 plants that produced CDM
(chlorophyll-deficient mutants) in their progenies. The survival rate did not decrease at lower than 15Gy irradiation with any LET values, and decreased as the
LET value increase at more than 20 Gy irradiation. In over all the seed fertility decreased as the dose increased, and there was no significant difference with increased
LET values. Mutation rates were low at any dose of irradiation with more than 80 keV/
µ m. The irradiations at 15Gy with 50–70 keV/
µ m were most effective for
CDM induction in both ions. These results show that increase of mutation rates is achieved by controlling LET in adequate dose irradiation. In this study, we isolated several mutant lines such as salt-tolerant, bronzing, earliness, high yield, virescent and dwarf. The trait of these mutants was fixed in M3 or M4 generation. Such a short developing period is a useful advantage of ionbeam treatment for breeding. Furthermore these mutants could be important as genetic resources for research in functional genomics of rice.
P0273 – ePoster
Plant hormones and male sterility in cereal crops caused by temperature stress
442
Higashitani, A 1
Watanabe, M 1
, Sakata, T 1 , Oshino, T 1 , Yano, K 2 ,
1 Graduate School of Life Sciences, Tohoku University,
Japan; 2 Meiji University, Tokyo, Japan
Plant reproduction processes are threatened by high temperature (HT) injury caused by recent global warming. Lobell and Field have reported that, at least in the cases of wheat, maize and barley, there is clearly a negative correlation between worldwide crop yields and increased temperatures, and for these crops, recent warming has resulted in an annual combined loss of approximately 40 megatons or $5 billion (Environ. Res.
Lett. 2007, 2: 014002). In particular, severe injuries by high-temperature stress commonly appeared during pollen development, and these result in male sterility.
Despite lots of previous histological and physiological studies, molecular mechanisms are not still clear. We therefore focused on entire gene expression during hightemperature injury in barley. Increasing temperatures repressed the expression of YUCCA auxin biosynthesis genes, and endogenous auxin levels specifically decreased in the developing anthers of barley. Moreover,
DNA proliferation in mitochondria, chloroplasts and nuclei of developing panicles is inhibited with increasing temperatures. Following DNA proliferation suppression, terminal abnormalities were observed in the organelles of anther wall cells, including mitochondrial swelling and over-development of chloroplasts. Comprehensive transcriptome analyses using both reproductive organs and vegetative tissues showed high and positive pairwise correlations between the expression profiles of auxin induced genes, DNA replication-related genes and mitochondrial-related genes. In contrast, the expression profiles of auxin-repressed protein genes and photosynthesis-/chloroplast-related genes were negatively correlated with those of the previously mentioned genes. Under HT conditions, the former was repressed and the latter was up-regulated in the developing panicles. Furthermore, application of exogenous auxin promoted the expression of DNA replication-related genes under HT conditions, inducing anther cell proliferation, and completely reversed male sterility. In addition, in the developing anthers of
Arabidopsis , a similar suppression of anther-specific auxin activity was observed with elevated temperatures, suggesting that the mechanisms underlying HT injury are highly conserved across plant species. These findings suggest that tissue-specific auxin reduction is the primary cause of high temperature injury, which leads to the abortion of pollen development with transcriptional alterations involved in nuclear and organellar DNA proliferation. Thus, the application of auxin may help sustain steady yields of crops despite future climate change.
P0274 – ePoster
A survey of bamboo mosaic virus and its modification to express foreign genes focusing on temperate strains
Hisamoto, Y 1 , Natsuaki, T 2 , Kobayashi, M 3
1 University Forest In Chiba, Graduate School of
Agricultural and Life Sciences, University of Tokyo,
Japan; 2 Dept of Bioproductive Science, Faculty of

Agriculture, Utsunomiya University, Japan; 3 Dept of
Forest Science, Faculty of Agriculture, Utsunomiya
University, Japan
Introduction: Bamboos form a huge rhizomatous clone with a gregarious flowering habit. Abandoned bamboo forests have spread unfavorably in Japan. To manage or clear those invasive bamboos, a novel biological technique is expected for controlling bamboo propagation. We isolated a flowering gene,
FLOWERING LOCUS T homolog, PmFT from
Phyllostachys meyeri to control flowering and seed production in bamboo. However, we were unable to introduce the isolated gene into bamboo clones for systemic gene expression. If the bamboo virus could be altered to a vector, the flowering gene would be distributed throughout the clone by the virus-modified vector through self-replication and movement. Bamboo mosaic virus (BaMV), a species of genus Potexvirus , is a single-stranded, positive-sense RNA virus that causes mosaic symptoms and reduces vigor. In the present study, we investigated BaMV infection in bamboos collected from Taiwan, Tanzania, and Japan, and tried to develop a BaMV vector for target gene expression in the entire bamboo clone.
Materials and methods: BaMV infections were investigated by observing symptoms and examined by
RT-PCR. Total RNA was extracted from 16, 16, and 30 bamboo leaves collected from Taiwan, Tanzania, and
Japan, respectively. Phylogenetic relationships among the BaMV strains were analyzed by comparing partial nucleotide sequences. Furthermore, full-length sequences of BaMV isolated from 3 bamboo species were determined to construct the vector.
Results and Discussion: Bamboo leaves with yellow mosaic symptoms were observed in major commercial bamboos in Taiwan and some culture collections in the
Fuji Bamboo Garden (FBG), Japan, but none in
Tanzania. BaMV infection was detected by RT-PCR in 9 bamboo species in Taiwan, of which 7 species were tropical and 2 temperate, i.e., Arundinaria usawai and
Pseudosasa usawai . A phylogenetic parsimony analysis indicated that BaMV strains isolated from temperate bamboos form a sister clade in the most advanced grade.
In FBG, BaMV infection was detected from 4 introduced bamboos; 2 each from tropical Asia and China (via the
USA). Tanzanian bamboos are also under inspection now. In general, minute leaf tissues compartmentalized by longitudinal and cross-veins in temperate bamboos are considered the unit of supercooling, and thus, temperate bamboos are suspected to exhibit virus resistance similar to cold hardiness, with blocking in minute tissues. The
6365 and 6364 nucleotide sequences of full-length cDNA from the BaMV strains BaMV-Bo and BaMV-Pu, isolated in Taiwan from Bambusa oldhamii and P. usawai , respectively, were determined. These sequences exhibited 7–10% nucleotide variations compared with other strains recorded in the Genbank database. The sequence of A. usawai from Mt Yangmingshan, Taiwan, is still under construction, probably because of a complicated 5 side-nucleotide sequence structure for
ORF1. The BaMV-Bo sequence was cloned into a pUC19-based plasmid vector between a cauliflower mosaic virus 35S promoter and a nopaline synthase terminator. The infective ability of the BaMV-Bo clone was confirmed when Nicotiana benthamiana leaves were infected by the particle gun method.
P0275 – ePoster
Vulnerability to branch xylem cavitation in
Prunus
x yedoensis
can predict tree decline
Ikeda, T 1 , Ueda, M 1 , Yanai, J 1 , Kasuya, N 1 , Ogura, K 2
1 Kyoto Prefectural University, Kyoto, Japan; 2 Kyoto
Botanical Garden, Kyoto, Japan
Introduction: Prunus x yedoensis Matsum., Yoshino
Cherry, is one of the most popular and widely planted cultivated flowering cherries (somei-yoshino) in temperate climates worldwide, especially in Japan. Those trees more than 50-year-old recently declined and died.
In response to this, tree doctors are trying to treat in diverse ways to fix them up. However. a preliminary diagnosis is not adequate, because it is judged by visual diagnosis, but not a judgment from a plant physiological viewpoint. Most of the declined P.
x yedoensis are infected with wood decay fungi, and show earlier defoliation, reduction of leaf biomass and dieback of shoots. Judging from those symptoms, declined trees seem to suffer from water deficit. In order to diagnose tree vigor accurately, it seems useful to evaluate water relations in P.
x yedoensis . In this study, we investigated following perspectives on water relations: (i) water relations parameters of leaves, (ii) xylem vulnerability to embolism of branches and (iii) soil survey such as soil chemistry, soil profile and soil hardness. Finally we examined the consistency of judgments between a visual diagnosis and a plant physiological one.
Materials: Fifty to eighty-year-old Yoshino Cherries, P.
x yedoensis , growing at the Kyoto Botanical Garden, Japan were used for our study. P.
x yoshino is a hybrid cherry.
All of P.
x yedoensis are clone because seedlings are propagated by means of grafting.
Methods: Measurements on water relations are as follows: (i) to evaluate water relations of leaves, midday leaf water potential and water relations parameters using the pressure-volume curve method were measured during summer and (ii) to evaluate vulnerability to xylem cavitation of branches, vulnerability curves were established using the air injection method. Studied trees were visually diagnosed and were discriminated between declined trees and sound ones. Visual reference was conformed to the tree doctor guidelines.
Results and discussion: There were not significant differences in midday leaf water potential, water relations parameters and vulnerability of xylem cavitation between visually declined trees and visually sound ones. Water relations of some visually declined trees were normal, meanwhile those of visually sound tree were bad. In other words, visual diagnosis did not necessarily correspond to physiological diagnosis. However, interesting results were obtained from analysis of water relations of individual trees. Tested trees were differentiated into three groups by vulnerability curves of branches, which were more vulnerable, less vulnerable and intermediate between more and less vulnerable.
Some of the more vulnerable group died next year and the year after. There was no problem with soil chemistry, but the soil was very hard due to tread pressure of visitors. Furthermore some root systems suffered from root rot. These seem to lead to water deficit in trees.
Water deficit for many years makes branch xylem more vulnerable to xylem cavitation caused by cavitation
443

fatigue. So, evaluation of vulnerability of branch xylem embolism can be possible candidate to predict tree decline and death.
P0276 – ePoster
Functional analysis of the gentian orthologues of the
FT/ TFL1 gene family
Imamura, T 1 , Nakatsuka, T 1
H 1
, Nishihara, M 1 , Takahashi,
1 Iwate Biotechnology Research Center, Japan
Gentian plants are a perennial plant and known as popular ornamental flowers. Since the flowering period of their cultivars is short and fixed between summer to autumn, the flowering control technique is required.
Unfortunately, there was no study about flowering initiation and environmental factor affecting flowering in gentian plants. To identify the machinery of the flowering initiation, we isolated and characterized the gentian FT / TFL1 family genes. First, we obtained three genes possessing high homology to FT / TFL1 and denoted GtFT1, GtFT2 and GtTFL1, respectively. The seasonal expressions of these genes in leaves and shoot apical meristem (SAM) were analyzed by a real-time RT-
PCR. The expressions of GtFT1 and GtFT2 in leaves were remarkably increased prior to the flowering initiation, whereas GtTFL1 was highly expressed at vegetative stage and was remarkably decreased just before the flowering initiation in SAM. We compared the expression pattern of GtFT / GtTFL1 family with earlyflowering and late-flowering lines. Both lines showed remarkable increase of GtFT1 and GtFT2 at the flowering initiation. The result suggested that GtFTs might play roles in the flowering initiation in gentian as well as other plants. However, the GtTFL1 expression was higher in late-flowering line than that in earlyflowering line at vegetative stage. To know why the different expression pattern of GtTFL1 was observed, we compared approximately 10 kbp region of GtTFL1 genomic DNA between early-flowering and lateflowering lines. We found that early-flowering line has a
320 bp insertion in promoter region. Moreover, we examined this sequence by genomic PCR in 14 gentian cultivars with different flowering times. The additional
320 bp sequence was found in the early-flowering lines, whereas no additional sequence was found instead in late-flowering lines. Cultivars with intermediate flowering times had both sequences. These results implying that the insertion may be related to the flowering time in gentian.
P0277 – ePoster
1
Starch structures/properties in high-amylose rice
Abe, K 1
S 5
, Hanashiro, I
, Itoh, K 6
2 , Idichi, K 3 , Sato, Ki 4 , Akuzawa,
Graduate School of Science and Technology, Niigata
University, Japan;
University, Japan;
2 Faculty of Agriculture, Kagoshima
3 Graduate School of Agriculture,
Kagoshima University, Japan; 4 Graduate School of Food and Nutritional Science, Tokyo University of Agriculture,
Japan; 5 Faculty of Applied bioscience, Tokyo University of Agriculture, Japan
Starch is major storage product of plant and is major sourse of carbohydrates for human. For the use of the starch in food industry, starch properties are good index, and are affected by glucan and starch granule structures.
The major component of the starch is amylose and amylopectin, former has linear structure of alpha-1, 4 glucan chain, and latter has alpha-1, 4 glucan unit chains and numerable branches by alpha-1, 6 glycoside linkage.
The structural characteristics of two components depend on action of different classes of starch-biosynthetic enzymes, including starch synthases, branching and debranching enzymes. Alteration of the enzymatic activities alters microstructures of the starch and starch granule, and could change the starch properties. In this study, we generated transgenic lines modified the expression of two starch biosynthetic enzymes in wx rice, and then analyzed the altered structures/properties of the starches. Line WAB expresses Granule-bound starch synthase I (GBSSI), resulted in synthesis of higher amount of amylose and extra long chain (ELC) in amylopectin(1), (2), and line WABEIIb lacks expression of Branching enzyme IIb (BEIIb), resulted in no major changes of unit chain distribution in amylopectin. We also generated line WAB/WABEIIb by intercrossing between line WAB and line WABEIIb. Line
WAB/WABEIIb showed production of high-amylose type starches, and major changes of unit chain distribution of amylopectin. The changes of structures and properties for these transgenic starches will be discussed. (1) Hanashiro, I et al. (2008); (2) I. Hanashiro,
I et al. (2009).
P0282 – ePoster
Application of gene silencing for control of plant parasitic nematodes
1
Jones, M 1
WA State Agricultural Biotechnology Centre, Murdoch
University, Perth, WA
Healthy roots enable a plant to make full use of available water and nutrients. Plant parasitic nematodes are a neglected but economically group if plant pests that damage plant roots and cause annual crop losses estimated at US$120 billion p.a. New resources, in the form of the complete annotated genome of the model nematode, Caenorhabdhitis elegans , and more recently genomes of two root-knot nematode and one cyst nematode, provide new information to identify new target genes in plant parasitic nematodes. Using these resources, a range of transgenic dicotyledonous and monocotyledonous plants (wheat and sugarcane) have been generated containing synthetic resistance genes to confer resistance to different nematode parasites.
Progress will be prevented on this research, including the results of nematode challenge experiments from two different approaches that show promise in conferring host plant resistance to economically important plant parasitic nematodes.
444

P0283 – ePoster
Ion-beam mutagenesis: New technology for the control of deletion-size by heavy-ion beam irradiation
Kazama, Y
Shirakawa, Y
1
1
, Hirano, T
, Hayashi, Y
1
1
, Nishihara, K
, Abe, T 1
1 , Ohbu, S 1 ,
1 RIKEN Nishina Center, Japan
Heavy-ion beams consist of ions accelerated to about
50% of the light speed, and have a high linear energy transfer (LET). While LETs of
γ
-ray and X-ray are 0.2 keV/
µ m and 2 keV
µ m, respectively, LETs of heavy-ion beams in the RIKEN RI-beam factory have a range from
22.5 keV/
µ m for C ion to 4000 keV/
µ m for Fe ion. This high LET irradiation is an excellent technology for inducing mutations to improve horticultural and agricultural crops with high efficiency. We previously reported that the LET value affects the mutation frequency, and that the most effective LET is 30.0 keV/
µ m for A. thaliana (Kazama et al. 2008). In
Mesorhizobium loti , symbiotic bacterium, irradiation of
Fe ion (640 keV/
µ m) showed a tendency to induce lager deletions than that of C ion (22.5–40.0 keV/
µ m) (Ichida et al. 2008). From the result, we postulate that the LET may also influence the deletion size in the mutated genes.
In the present study, therefore, we aim at developing a novel method for deletion-size-controlled mutagenesis by irradiation of heavy-ion beam with different LETs in plant mutagenesis. As a first step toward controlling deletion size, we investigated the LET effect on induced mutations in Arabidopsis thaliana . Dry seeds of A. thaliana (Col-0) were irradiated with C (22.5 keV/
µ m,
30.0 keV/
µ m, and 290 keV/
µ m), or Ar (290 keV/
µ m).
For detection of mutations, morphological mutants of which responsible genes have been well characterized were screened in M2 generation, and then responsible genes of the mutants were sequenced. We analyzed the mutations in responsible genes of the 48 mutant lines.
Mutation sites on the genes contained base substitution, deletion, insertion, and chromosomal rearrangement. Of the detected mutations, 41 had deletions in their responsible genes, of which 24 were simple deletions and the others were complicated deletions accompanied by a translocation or base substitutions. With 22.5- and 30.0keV/
µ m irradiations, 92% of the deletions were less than
100 bp. On the other hand, with 290-irradiation, such small deletions were only 40%, and the other deletions were larger than 100 bp. These results suggest that deletions induced by heavy-ion beam have a tendency to increase the size with increasing LET. Additionally, such large deletions can be detected by a whole-genome tiling array, which allows us to investigate the number of deletions par genome in the heavy-ion induced mutants.
Data of the whole-genome tiling array will be present in the conference.
P0284 – ePoster
Micropropagation of
Satureja khuzistanica
Jamzad., a potential source of Carvacrol
Ramak, P 1 , Sharifi, M
Ebrahimzadeh, H 2
1 , Kazempor osalo, S
, Ahmadian, N 1
1 ,
1 Tarbiat Modares University, Tehran, Iran; 2 Tehran
University, Tehran, Iran
Satureja khuzistanica Jamzad (Persian name, ‘marzeh khuzistani’) of the family Lamiaceae is an endemic plant of Iran that is widely distributed in the southern part of
Iran. This species belongs to the family Lamiaceae, subfamily Nepetoideae and the tribe Mentheae and close to Satureja edmondi Briquet but with some differences in stem, leaf and inflorescence (Jamzad ,1994). The essential oil of Satureja khuzistanica Jamzad is characterized by high concentration of Carvacrol (93%).
To date there have been no reports on the propagation of this species either conventional or non-conventional methods. In this context, we report a protocol for micropropagation and rooting of this important aromatic plant. Node segments excised from in vitro seedling and established on LS, B5, MS and half-strength MS medium. The highest propagation rate was obtained using LS medium supplemented with 1 µM 6benzylaminopurin (BAP). For shoot proliferation, The node explants were cultured on LS medium supplemented with different concentrations of BAP and kinetin (Kn) within the range of 0.5 to 2 µM and combinations of indole-3-butyric acid (IBA: 2 and 5 µM) with BAP at two concentrations (2 and 5 µM). Multiple shoots were obtained from the nodal explants, the higher frequency (77%) formation of shoots was observed in the
LS media that contained BAP (5 µM) in combination with IBA (2 µM).The best condition for rooting were Ls medium plus 2.5 µM of indole-3-butyric acid. The rooted plants were successfully transferred to garden soil, exhibiting a normal development. Key words: Satureja khuzistanica , node explants, 6-benzylaminopurin, growth regulators, micropropagation Reference Jamzad,Z., A new species of the genus Satureja (Labiatae) from Iran,
Iran J. Bot. 6 (1994) 215–218.
P0286 – Poster
Response of Anise (
Pimpinella anisum
L.) to organic and natural inputs
Khalesro, S 1 , Ghalavand, A 1 , Sefidkon, F 2
1
Asgharzadeh, A 3 , Mahdavi, B 1
Tarbiat Modares University, Iran; 2
,
Research Institute of
Forests and Rangelands, Tehran, Iran; 4 Soil and water
Research Institute
Biofertilizers and organic manure not only have a positive effect on growth and yield of plants, but also provide the safety of environment. Accordingly, in this research the effects of PGPR, vermicompost and natural zeolite on growth, yield and essential oil composition of
Anise were investigated. Anise ( Pimpinella anisum L., family: Apiacea) is one of the most important aromatic and medicinal plants that’s essential oil is of great value and has vast application in medicinal and food industries.
Field experiment was conducted in the Agriculture
Research Station at Sanandaj, Iran, in 2008. Plants were treated with three levels of vermicompost (0, 5 and 10 t.ha
-1 ), two amounts of zeolite (0 and 4.5 t.ha
-1 ) and a mixture of Azotobacter chroococum, Azospirillum lipoferum and Pseudomonas fluorescens (inoculation and not-inoculation). Experimental design was randomized complete blocks design with factorial arrangement of
445

treatments with three replications. The results showed that vermicompost levels significantly increased plant height, 1000 seed weight, biological yield, seed yield, essential oil content, anethol and methyl cavicol percentage. The highest amount of them was obtained from the maximum rate of vermicompost. Inoculating
Pimpinella anisum seeds with PGPR led to significant increment for all of mentioned characters except for 1000 seed weight and methyl cavicol percentage. Zeolite application had significant effect on plant height, biological yield, seed yield and essential oil content but no significant effect on the other characters including
1000 seed weight, anethol and methyl cavicol percentage. In addition the highest values for biological yield (3005 kg. ha -1 ) and seed yield (1099.15 kg. ha -1 ) were obtained as a result of the maximum rate of vermicompost (10 t.ha
-1 ) combined with PGPR inoculation. Finally, application of vermicompost, PGPR and zeolite have beneficial effects on growth and yield of
Anise.
P0287 – ePoster
Improvement of wheat and barley frost tolerance by constitutive and cold-inducible overexpression of
DREB/CBF transcription factors
Kovalchuk, N
Pyvovarenko, T
Langridge, P 1
1
1
, Jia, W
, Shirley, N 1 , Ismagul, A 1
, Lopato, S 1
1 , Eini, O 1 , Bazanova, N 1
, Singh, R 1
,
,
1 ACPFG, Australia
Cold (freezing temperatures) is among the major environmental factors limiting crop productivity worldwide. Vegetative tolerance is a concern in parts of the world where cereal crops need to overwinter at the vegetative stage under severe conditions (e.g. -20 ° C) before flowering in the spring/summer. The direct damage caused by frost in Australia (around 100 M/yr) may be modest compared to that caused by drought or salinity, but an overnight frost event in the order of -2 to
- 8°C during the flowering stage can damage the sensitive reproductive structures and cause near-total loss of grain set. We have recently demonstrated considerable improvement of the vegetative frost tolerance of transgenic barley with strong and moderate constitutive up-regulation of TaDREB2 and TaDREB3 transcription factors (Morran et al. 2011). However, improvement of frost tolerance is accompanied with several undesirable changes in plant development, including slow development and delay of flowering. These problems could be overcome by using cold-inducible and/or spikespecific promoters. Therefore, we have developed several cold inducible promoters with different levels of strength and basal activity. We will present preliminary results of the analysis of plant development and vegetative frost tolerance for two transgenic populations of wheat and barley, with elevated levels of the cold tolerance gene
TaDREB3 regulated by strong and weak cold-inducible promoters. An assessment of the tolerance of vegetative organs of 2x35S-TaDREB2 and 2x35S-TaDREB3 transgenic barley plants is being extended to reproductive tissues. The T3 generation of plants with normalised/partially normalised developmental phenotypes were analysed for the frost tolerance at the reproductive stage of development.
446
P0290 – ePoster
A physiological investigation on the response of sugarcane to water stress
Basnayake, J
N 3
1 , Jackson, P 2
, Lakshmanan, P 4
, Inman-Bamber, G 2 , Berding,
1 BSES Limited, Ayr, Australia;
Townsville, Australia; 3
2 CSIRO Plant Industry,
BSES Limited, Gordonvale,
Australia; 4 BSES Limited, Indooroopilly, Australia
With recurring drought and the large requirement of irrigation, efficient use of water and drought adaptation are becoming major determinants of sugarcane production in Australia. Nearly 60% of the Australian sugarcane production receives full or supplementary irrigation, and the annual economic loss due to water stress is estimated to be $261 million. Despite such a large economic impact, improving cane productivity under water-limited condition has never been specifically addressed through breeding in Australia. Towards that objective, we investigated the phenotypic and genetic variations of traits that are linked to or known to confer adaptation to water-limited production conditions in a genetically diverse sugarcane population, and their contribution towards productivity. Field experiments were conducted under rainfed, fully irrigated and imposed drought conditions at three locations in the sugarcane growing regions for three years. Substantial genotypic variation for cane and sugar yield was observed under water stress conditions and the clones showed different degrees of susceptibility for cane and sugar yield. Among the different traits studied, mid-day stomatal conductance (gs), canopy temperature (CT), leaf temperature (LT) and relative water content of leaf tissue
(RWC) showed significant genotypic variation under water-limited conditions. However, gs and RWC were the most useful traits to discriminate clone performance under water stress as they were stable across different locations and crop cycles, and showed significant phenotypic and genetic correlation with cane yield and sugar production under drought. Based on the physiological and yield attributes four groups of clones were identified. Multivariate analyses showed a significant contribution of gs, RWC and the potential yield for clustering those clones. The physiological mechanisms underpinning those groupings from a crop adaption and productivity perspective will be discussed.
Key words: sugarcane, drought, adaptation, phenotypic variation, genotypic variation.
P0292 – ePoster
Phytoremediation of organic contaminants in soil: survey of species and associated techniques for future
1 applications
Lima, AL 1
Marcio AR 1
, Leite, DC 1 , di Romagnano, LFT 1 , Nahuz,
Center for Forest Resource Technology - Institute for
Technological Research/IPT
The state of São Paulo has the largest number of contaminated areas and the highest levels of contamination of Brazil, exposing more than 500.000

inhabitants to toxic agents. Contamination by organochlorine compounds widely used in agriculture and policies of public health, such as herbicides and pesticides, outweigh those derived from petroleum, industrial waste and heavy metals on a national scale.
Even São Paulo, which has as main pollutants aromatic solvents, fuels and polycyclic aromatic hydrocarbons, presents critical situations of pesticide contamination.
The impacts caused by these contaminants are harmful to society and the environment, since in addition to high toxicity, they are persistent, accumulating along food chains.The search for solutions to remediate contaminated areas by organochlorines has been widespread through various physical, chemical and even biological technologies. Biological techniques are concentrated in using of microorganisms and/or vegetation and are proving to be very promising for the decontamination of organochlorine compounds in soils and groundwater, since they favor and stimulate natural mechanisms leading to removal and degradation. These techniques even amplify the recovery of landscape and represent relative low investment. Recent research and international cases have demonstrated great efficiency with the use of several plant species, associated directly or indirectly to microorganisms in the remediation of contaminated areas. Nevertheless, knowledge and application of this technology are still scarce in Brazil.
We surveyed phytoremediator plant species for organochlorine compounds and those occurring on contaminated areas (for being tolerant in contaminated environments), to generate a database to support future research on phytoremediation. The main scientific databases were consulted between April 2010 and
December 2011; these being Web of Science, SciELO,
Highwire, Scirus, ScienceDirect, Scopus and LILACS, in addition of the collective information from universities, institutes and companies. We found 164 species, belonging to 47 plant families, with phytoremediation action demonstrated and 38 species (from 35 families) with potentiality for phytoremediation. The families that stood out were Poaceae (grass) and Fabaceae (legumes), with species and genera that occur in the edaphoclimatic conditions of São Paulo. Complementary techniques, associated directly or indirectly to phytoremediation were outlined for evaluation. The intensification of the processes of natural degradation, e.g. by applying organic matter improves soil quality, increases microbial colonization, promoting more efficient and faster organochlorine degradation.
P0293 – ePoster
The study and genetic improvement of main warm season turfgrass conducted in China
Liu, J 1 , Guo, H 1 , Guo, A 1 , Zong, J 1 , Xuan, J 1 , Chen, J 1
1 Institute of Botany, Jiangsu Province and Chinese
Academy of Sciences, China
With the aim to nationalize the Chinese turfgrass industry, a lot of work have been conducted on turfgrass germplasm resources collection, assessment, genetic improvement and extension since 1994. On the basis of these works, 1300 accessions of warm season turfgrass genetic resources, belonging to 20 species of 10 genus, were collected and introduced from domestic and abroad, and establish the most extensive turfgrass gene bank in
China. Through the systematical study on morphological variation, reproductive performance and stress resistance
(cold tolerance, drought resistance, salinity resistance and aluminium tolerance) of turfgrass genetic resource(mainly including bermudagrass, zoysiagrass and centipedegrass), we innovatively proposed a set of novel index and evaluation system, including the turfgrass creeping characteristics index, the apparent permanent wilting index and the comprehensive assessment system of turf quality, which has been widely used in China; we had clarified the inheritance mode of the important vegetative traits, reproductive traits and cold tolerance of important warm season turfgrass; screened the closely linked molecular markers with cold resistance, green period and salinity resistance traits; preliminarily constructed a high-resolution centipedegrass molecular genetic mapping. Cold tolerance related transcription factor gene ZjDREB1 and actin gene ZjACT were also firstly cloned from zoysiagrass and successfully expressed in prokaryote and eukaryote organism. Using systematic breeding, cross breeding, radiation induction and somatic mutation techniques, 8 bermudagrass cultivars (lines), 19 zoysiagrass cultivars (lines), 13 centipedegrass lines were bred, and a set of somatic mutation lines of zoysiagrass and centipedegrass with cold and drought resistance were obtained. Our research work has been highly valued. from 1994 on, more than 150 papers and 5 books were published; the first and third prize of science and technology awards at province-ministry level were honored, respectively; 3 cultivars including Nanjing bermudagrass, Yangjiang bermudagrass and Suzhi No.1 hybrid zoysiagrass were registered at the state level and
15 varieties were identified at province-ministry level; 2
Chinese invention patents were granted. The extension area of the released cultivars were more than 5 million square meters in total.
P0296 – ePoster
1
Molecular characterization of somaclones using
RAPD marker In patchouli
Masgod, G 1 , Behera, A 1
Bangalore University, India
The plants regenerated through indirect organogenesis from leaf explants of Pogostemon cablin Benth. variety
Johor , an essential oil yielding plant showed variation with regard to morphological characters- five tall plants with few branches and thick leaf; five tall plants with more branches and broad leaf and six dwarf plant with more branches and normal leaf, among 100 in vitro plants sreened. Therefore, Randomly Amplified
Polymorphic DNA (RAPD) analysis was employed to assess genetic fidelity among isolated somaclones. The selected sixteen plants of SC5 (SC1 first generation after an in vitro phase followed by SC2-SC5) generation were subjected to RAPD analysis. Out of 8 random 10-mer primers, 5 primers generated polymorphism. A total of scorable 649 bands ranging from 100bp to 3kbp were amplified. Further, among the sixteen plants, five plants revealed 38 polymorphic bands,being 5.8 per cent of the total bands. The result demonstrates that RAPD markers can be used successfully to determine the genetic fidelity
447

and confirms that the isolated somaclones are morphologically and genetically different from that of the control plants.
P0297 – ePoster
Contribution to the chemistry of
Croton
L.
1
(Euphorbiaceae)
Matos, L 1
ML 1
, Salatino, A
, A.C. dos Santos, D
Motta, L 1 , Cordeiro, I 3
São Paulo, Brazil; 4
1 , Negri, G
1
Federal de São Paulo, Brazil; 3
2 , Faria Salatino,
, Furlan, C
Universidade de São Paulo, Brazil;
1
, Ribes de Lima, L
, Barbosa da
2
4
Universidade
Instituto de Botânica de
Universidade Federal de São Carlos,
Brazil
Croton L. is one of the largest genera of Euphorbiaceae, comprising 300 species in Brazil. Plants of this genus are widely used in popular medicine to treat wounds, inflammatory diseases, gastrointestinal and hepatic disturbances, diabetes, hypertension and even cancer.
The variety of medicinal applications reflects the diversity of biologically active compounds, mainly alkaloids, flavonoids, essential oils, diterpenes and triterpenes. The present study focuses on the chemical screening of seven Brazilian Croton species ( C. betulaster Muell. Arg., C. glutinosus Muell. Arg., C.
antisyphiliticus Mart., C. hemiargyreus Muell. Arg., C. grandivelum Baill., C. pycnocephalus Baill., C. montevidensis Spreng.) by means of chromatographic and spectrometric (GC/MS and HPLC/MS) analyses of essential oils, flavonoids and compounds of crude ethanolic extracts. The essential oils are rich in mono and sesquiterpenes, mainly alpha-caryophyllene. No phenypropanoids were found. Tiliroside, a bioactive kaempferol acylglycoside, is a major constituent in flavonoid fractions. Oxygenated sesquiterpenes (e.g. caryophyllene oxide), steroidal triterpenes (e.g. sitosterol), N-methyl-crotonosine and pronuciferine, benzylisoquinoline-like alkaloids typical of Ranunculales and common in Croton , were predominant in the EtOH crude extracts. The present study provides the first data about the chemistry of the studied Croton species, apart from the presence of alkaloids and clerodane diterpenes in C. hemiargyreus and the tetramethoxylated flavonol casticin in C. betulaster , which were already reported in literature. The data obtained suggest pharmacological potentials and might be used as chemotaxonomic tools at the infrageneric level.
P0298 – ePoster
Assessment of variability in mothbean germplasm
(
Vigna aconitifolia
) and
V. trilobata in the Indian arid region
Meghwal, RR 1 , Dwivedi, NK 1
1
GS 3
, Yadav, S 2 , Krishnan,
NBPGR, RS, Jodhpur, India;
New Delhi, India; 3
2 NBPGR, Pusa Campus,
Division of Genetics, IARI, Pusa
Campus, New Delhi, India
Mothbean ( Vigna aconitifolia (Jacq.) Marechal) is a traditional legume cultivated by traditional farmers in
448 arid region of India. It is a minor Kharif pulse crop and considered as one of the most adapted arid legume in the region, because of its tolerance to drought and heat. It is cultivated sporadically in the dry habitats and grown as a pasture, fodder, and green manure crop. The mat like growth of the plants makes it superior to other arid legumes such as cow pea, cluster bean. It is one of the important sources of proteins, calories, essential amino acids (lysine and lucine), minerals for the poor as well as rich people in arid and semi-arid regions of India and abroad. Vigna trilobata (Chidi moth) the wild relative of mothbean endemic to India is considered as progenitor of the species. Adak moth is considered a weedy form of
Vigna aconitifolia . Variability for eighteen promising accessions of V. aconitifolia (cultivars) along with three checks, eight accessions of weedy form of V.
aconitifolia and eight accessions of V. trilobata were assessed during
Kharif 2008- 09. These accessions were characterized for
11 qualitative and 18 quantitative characters at National
Bureau of Plant Genetic Resources, Regional Station,
Jodhpur, India. A wide range of variability was observed for qualitative characters such as days to germination, early plant vigor, plant growth, plant growth habit, leaf color, leaf pubescence, days to first flowering, days to
50% flowering, days to first and 80% maturity, pod color at maturity, seed shape and seed color. Variability among quantitative characters namely plant weight, plant height, peduncle length, number of branches per plant, no. of pods on branches, cluster length, no. of clusters on branches, no. of clusters per plant, pod length, pod yield per plant, no. of seeds per pod, seed yield per plant and
100 seed weight were recorded. Variability was also recorded in protein content in dry seeds of V. aconitifolia and V. trilobata . The protein content in promising accessions of V. aconitifolia ranged from 22.86–27.33%, while 21.74– 25.50% and 22.34– 25.0% were recorded in weedy form of V. aconitifolia and V. trilobata respectively. The accessions number IC329044,
IC329037, IC16221 and IC311396 were identified as more potential genotypes. On the basis of present investigation it is concluded that the diversity and variability studied in mothbean with their weedy (Adak moth) and V. trilobata (wild) are useful tools in germplasm management and future planning of any improvement and breeding programmes related to this crop.
P0299 – ePoster
Euclidean cluster analysis in Mothbean (
Vigna aconitifolia
(Jacq.) Marechal) germplasm in arid
1 region of India
Meghwal, RR 1 , Dwivedi, NK 1
NBPGR, RS, Jodhpur, India; 2
, Mathur, BK 2
CAZRI, Jodhpur, India
Euclidean cluster analysis was assessed in one hundred and eight accessions of Mothbean ( Vigna aconitifolia
(Jacq.) Marechal) germplasm along with three checks namely, Jadia, Marumoth and RMO-40 using D2 statistics. These germplasm accessions were grown at
NBPGR, Regional Station, Jodhpur, India during 2006 in augmented randomized complete block design for high yielding, desirable plants. The observations were recorded on five randomly selected plants in each row for
18 quantitative and 11 qualitative characters. All the

germplasm lines were classified into eleven clusters.
Cluster V was the largest comprising of 20 genotypes followed by cluster IX with 19 genotypes. The genotypes of cluster II had maximum intra cluster distance (20.426).
The maximum inter cluster distance was recorded between cluster II and cluster XI (116.10). The clustering pattern of the strains, in general, did not follow their geographical distribution. Based on cluster means the important clusters are viz., cluster I for superior performance of no. of branches per plant (50.84) while cluster II superior for cluster per plant (63.47), cluster length (2.54), pods on branches (156.0), pod yield per plant (24.35) and seed yield per plant (18.32). An effort has been made to collect and screen mothbean germplasm for high yield and resistance to disease.
P0300 – ePoster
Oil yields and fatty acids distribution of pericarp and seeds of
Oenocarpus bataua and
O. bacaba
: two potential Amazonian palms for biodiesel production
Meyer, JM 1 , Salatino, A 1
1 Universidade de São Paulo, São Paulo, Brazil
Due to depletion of fossil fuels and environment degradation, alternative fuels represent a relevant subject in scientific and technological research. In Brazil, the
Agriculture Ministry supports biodiesel production from oleaginous plant sources. Prospective studies of native oleaginous plants as oil feedstock for biodiesel production may contribute to improve social and economic conditions of certain Brazilian regions, especially North and Northeast. Arecaceae are potential oil sources because of the frequent high yields of their species. Thus they may represent an alternative for sustainable development. This present study aimed to determine the lipid content and the fatty acids distribution of dried pulps and kernels of Oenocarpus bataua and O. bacaba grown naturally in the Amazonian
North region of Brazil. The percentage of lipids determined by Soxhlet extraction with hexane was 1.5% in kernel and 30.7% in pulp, regarding O. bataua , and
1.2 and 19.7%, regarding O. bacaba . The fatty acids were analyzed by gas chromatography. Saturated acids of the kernel oil of O. bataua were capric (9.1), lauric (4.0), myristic (1.5), palmitic (16.8) and stearic (2.7), while the unsaturated acids were oleic (19.4), linoleic (36.5) and linolenic (4.2). The saturated acids of the pulp oil were myristic (1.3), palmitic (19.5), and stearic (2.3) and the unsaturated acids were palmitoleic (1.1), oleic (46.5), linoleic (19.5) and linolenic (2.3). Regarding O. bacaba , saturated fatty acids of the pulp were myristic (3.7), palmitic (27.8), and stearic (3.8), and the unsaturated acids were palmitoleic (1.15), oleic (26.8) and linoleic
(25.12); in kernel oil, the saturated acids were myristic
(2.8), palmitic (16.2) and stearic (2.9), and the unsaturated acids were oleic (15.1) and linoleic (22.3).
Kernel and pulp fatty acid compositions of both species are similar to palm oil ( Elaeis guineensis ), widely used in
India and Malaysia for biodiesel production. Though physical studies of the oil are still necessary, O. bataua and O. bacaba are seemingly potential oil sources for biodiesel production.
P0303 – ePoster
Radiation mutagenesis: molecular characterization of
1 mutations induced by gamma irradiation and heavyion beam in rice
Morita, R 1 , Hayashi, Y 1
Nishimura, M 2 , Abe, T 1
, Shibukawa, T
RIKEN Nishina Center, Wako, Japan; 2
2 , Takehisa, H
NIAS, Japan
2 ,
Mutants are indispensable and powerful tools used in the analysis of plant gene function. Higher plant mutants are usually produced using T-DNA or transposons, chemical mutagens, or ionizing radiation. The insertion of T-DNA or transposons destroys gene structure. Chemical mutagens such as ethyl methanesulfonate or N-methyl-Nnitrosourea mainly cause base substitution by DNA base alkylation. Ionizing radiation produces both single and double strand DNA breaks. As a result, ionizing radiation has been thought to have the potential to cause various types of mutation. Ionizing radiation can be divided into two classes according to differences in linear energy transfer (LET) (the energy transferred per unit length of the track). Alpha particles, neutrons, and heavy-ion beam have high LET, while gamma rays, X-rays, and electron beams have low LET. However, there is almost no comprehensive information about mutations induced in plant genomes by both high and low LET ionizing radiations. To investigate mutations induced by ionizing radiations in higher plants, we first irradiated rice with gamma ray, screened mutations expressing five mutation phenotypes, i.e., glutinous endosperm (wx), chlorophyll b deficiency, endosperm protein deficiency, gibberellinrelated dwarfism, and shortened plastochron. We analyzed 24 mutants and identified the type and size of the mutations. Nucleotide sequence analysis showed that the most frequent mutation induced by gamma rays was deletion, particularly small deletion. Of the 24 mutations,
15 were small deletions (1–16 bp), four were large deletions (9.4–129.7 kb), three were single-base substitutions, and two were inversions. Deletions 100 bp–8 kb in length were not found, suggesting that gamma irradiation is unlikely to induce deletions of 100 bp to 8 kb but is more likely to induce deletions between 1 and several ten bp or those of around 10 kb or more. Then we irradiated rice with heavy-ion beam, collected mutants in order to clarify types of mutations. Based on the results, reverse genetics applications may be possible for ionizing radiation-induced deletions in rice.
P0304 – ePoster
Effect of aqueous extracts of some weeds on germination and seedling growth of tomato
1
(
Lycopersicon esculentum
Mill.)
Mukhtar, I 1 , Bajwa, Rukhsana 1
Institute of Plant Pathology, Pakistan
Tomato ( Lycopersicon esculentum Mill.) is the most important vegetable crop in Pakistan. Seed priming has been adapted in many developed countries to improve seed performance, through increased seed vigor subsequent to enhanced percentage of seed germination and rates of seedling emergence. The effect of seed
449

priming on germination percentage and mean time to germination of local cultivars of tomato ( Lycopersicon esculentum Mill.) was studied. Effect of aqueous leaf extract of some weeds Convolulus arvensis L.,
Euphorbia helioscopia L., Vicia sativa and Anagallis arvensis L. was assessed for their effect on the germination and seedling growth of tomato in petri-dish and in the field. Seeds of tomato were primed with aqueous extracts (0%, 10%, 20%, 30%, 40% and 50%) of the four weeds. Seeds were air dried on metallic mesh and were placed in petriplates lines with two layers of
Whatman filter paper soaked in sterile distill water. In each petriplate, ten seeds were placed. Plates were incubated at 25°C under dark. Germination percentage and germination index of tomato seeds was calculated.
Weed extract priming increased germination rate and germination percentage as compared to control. An interaction effect on germination performance between vigor and priming was observed. 30% of all tested weed extract, showed increased germination performance in seeds. Extracts of C. arvensis , E. helioscopia , V. sativa and A. arvensis showed increased germination percentage and growth enhancement in tomato seedlings.
In field, root and shoot length of seedling was also high overall at 30%, but, 20% A. arvensis extract showed significant increase in shoot and root length in field experiment.
P0305 – ePoster
Study of effects of late season drought stress in wheat cultivars using stress susceptibility, tolerance indices
1 and canopy temperature depression
Nazemi, G 1 , Dehghani, G 2 , Nazemi, S 2
2
Islamic Azad University, Haji Abad Branch, Iran;
Education Office Fasa, Iran
Drought is a wide-spread problem seriously influencing wheat production and quality. This research was done in two separate projects to determine drought tolerance of wheat cultivars and lines for selecting drought tolerant varieties using stress susceptibility, tolerance indices in
Darab, Iran during 2009-2010. This experiment was conducted with 25 lines and cultivars in a randomized complete block design with three replications under normal and terminal drought environments terminal.
Irrigation was done in normal conditions based on water requirements until physiological maturity, and in drought stress conditions irrigation wasn’t done after anthesis to physiological maturity stage. Based on combined analysis most traits in the genotypes showed a significant reaction to irrigation treatments. While, in some traits interaction of genotype and environment was not significant. Lines No 15, 2, 24, 10 and 12 have had the highest yield in drought environments respectively. In normal environment, lines 13, 17, 16, 2 and 4, have had the highest performance respectively. Comparison of means with Duncan method in combined analysis also showed that the lines 13, 15 and 17 have higher average yield than other varieties. Drought resistance indices showed that except for TOL and SSI in drought environment all indices have a significant correlation with yield in both environments. lines 13 and 15 with the highest value of STI, MP, and GMP and having the highest yield in drought environment had a good
450 resistance to drought. Calculating three times cultivar
CTDs from the flowering to maturity indicated that cultivars canopy temperature depression have a positive and significant correlation with yield. Average three times CTD in both environments also showed significant positive correlation with the yield.
P0306 – ePoster
1
Ornamental plant resources of
Hedychium
in China
Nian, L 1 , Xiu, H 1
Zhongkai University of Agriculture and Engineering,
China
Hedychium K. occurs in tropical and subtropical areas, it belongs to the plants of Zingiberaceae. There are about
50 species in this genus in the world. As a genus,
Hedychium is characterized by the conspicuous ligules, terminal inflorescences, very long (rarely short) filaments, petaloid staminodes and fragrant flowers. It is an economically important genus used for cut flowers, with species possessing flowers with a wide range of color and as fragrances such as honeysuckle, jasmine, sweet-scented osmanthus , or orchid. The wild existence state included species, distribution, habitat, and ornamental resource of Hedychium in China, are systematically researched by consulting the specimens and literatures, collecting and investigating the living species in the wild, and cultivating them in garden. The results shows that: (1) In China, there are 32 species, 2 varieties and 1 form, a total of 35 taxa. Of them, we found: 2 new species: H. longpetalum X.Hu & L.Nian and H. menghaiense X.Hu & L.Nian, 1 new form: H. villosum f. albifihamentum X.Hu & L.Nian, and 2
Chinese new record species: H. gardenarium and H. ellipticum . There are 18 special species in our country. H. bipartitum was reduced to H. flavum by us. We support
Tom Wood’s view: reduced H. puerense to H.
stenopetalum . And, we support Wu Telin’s view: reduced H. emeiense to H. flavescens . H.
efilamentosum was reduced to H. wardii by Fischer, we support this view. (2) This genus is mainly distributed in the southwest of China, from 50 m to 2500 m altitude. Some species are distributed in limestone area. Some species can grow in water. Most of them like to grow in the forest edge. (3) Most species of this genus in China are very beautiful ornamental plants, especially as a cut flower. Some species can be used as garden ornamental plants. (4) In order to easily identify and use those ornamental plants, this genus is divided into 6 groups:
Hedychium villosum group, H. coranarium group, H. yunnanense group, H. dengsiflorum group, H. coccineum group and H. yungjiangense group, based on their morphologic characters, distribution and wild environment characters etc.
P0307 – ePoster
Adaptation of wheat to climate change in southwestern Australia
, Bramley, H 23 , Siddique, KHM 3
1
Oliveira, E 1 , Palta, JA 2
A School of Plant Biology, University of Western
Australia, Crawley, Australia; 2 CSIRO Plant Industry,

Wembley, Australia; 3 Institute of Agriculture, University of Western Australia, Australia
Wheat ( Triticum aestivum ) is Australia’s most important grain crop and Western Australia production occurs where water supply is the most critical limiting factor to crop growth and yield. Climate change models predict that rainfall in Western Australia will decrease by up to
15%, evapotranspiration will increase and mean temperature will rise by 3–4°C, all relative to that in
1990. Based on existing physiological knowledge, there is a reasonable expectation that these changes will affect wheat production. However, the effects of elevated temperature and reduction rainfall combined with elevated [CO elevated [CO
2
2
] are unknown.We hypothesized that
] will mitigate the interactive effects of high temperature and terminal drought on growth and yield, and improve water use efficiency in wheat. We aim to determine how the interaction between elevated
[CO
2
], high temperature and terminal drought affects growth, yield grain protein content and size, and crop water use. Field experiments are been conducted at
UWA Field Station in Perth, WA using four naturally lit tunnels 10m length x 2m wide x 2.5m high. One tunnel has been set as control under ambient [CO
2
] and ambient temperature while the other three tunnels are been set at
700 µmol mol -1 [CO
2
] and 2, 4 and 6°C above ambient temperature, respectively. After 50% of flowering, water will be with-held from half of the crop in each tunnel to induce terminal drought.
P0308 – ePoster
Phytochemical screening and in vitro evaluation of antidermatophytic activity of
Oxalis corniculata subjected to various drying treatments
Pandith, J 1 , Singh, G 2
1 Dept of Botany, Govt. Degree College, Sopore,
Kashmir, India; 2 Dept of Botany, St. John's College,
Agra, U.P India
Purpose: the research on the medicinal plants should be extended with the identification of the active principles in the plants. Scientific examination of the remedies could lead to standardization and quality control of the products to ensure their safety. It is after such evaluations that they can be approved for use in the primary health care. Such research activities could also lead to the development of new drugs as in the past. Methodology: the present study was carried out to determine phytochemical analysis of
Oxalis corniculata subjected to various drying treatments and monitored the change in the chemical composition and antidermatophytic activity against human pathogenic fungi. Results: results obtained revealed that the drying treatments inhibits the number of phytocompounds and the maximum inhibition was observed due to oven dry at
70 ± 1ºC for 5 hours using ethanol as solvent, however the plant extract using ethanol as solvent by freeze drying for 2 hours inhibits 86.15 % in the mycelial growth of
Microsporum gypseum . The least 21.5% inhibition has been observed using acetone as solvent by Oven dry at
50± 1ºC for 9 hours in the mycelial growth of
Trichophyton mentagrophytes . Conclusion: it is interesting to note deviation in inhibitory effects due to various drying treatments which monitored the presence of phytocompounds present in selected plant material.
Hence plant drying treatments had a bearing in monitoring therapeutic potentiality of selected plant species against experimental organisms.
P0310 – ePoster
Photochemistry, photosynthesis and antioxidative defence capacity of two wheat cultivars with contrasting tillering ability grown in FACE
Posch, S 1
Norton, R 3
, Seneweera, S
, Fitzgerald, G 4
1 , Loew, M
, Tausz, M 2
2 , Dempsey, R 2 ,
1 University of Melbourne, Dept of Agriculture and Food
Systems, Australia; 2 University of Melbourne, Dept of
3 Forest and Ecosystem Science, Australia;
Plant Nutrition Institute; 4
International
Victorian Dept of Primary
Industries, Australia
According to current projections of global climate change, the atmospheric CO
2
concentration [CO
2
] is increasing and is expected to reach 550 micromol mol -1 by the middle of this century. Changes in [CO
2
] affect photosynthetic processes in plants. In general, plants respond to increases in CO
2
by increasing photosynthetic net assimilation rates (A) and internal CO
2 concentrations (Ci) as well as decreasing stomatal conductance (gs). Such changes can feed back on underlying processes of photosynthesis such as effective quantum yield of photosystem II (PSII, phiPSII), maximum quantum yield of light adapted PSII (Fv’/Fm’) and photochemical quenching (qP) as well as related antioxidative defence processes. For example, under elevated CO
2
(eCO
2 destined for CO
) photosynthetic electron transport assimilation will increase
2 proportionally with increasing A, while perhaps decreasing electron flux to oxygen, which would imply a decreased requirement for antioxidative defence. Wheat is counted among the three major cereal crops; it provides one fifth of the total calorific input of the world’s population. To secure future global food supply in a changing climate, it will be necessary to adapt current cultivar selection and breeding strategies to an eCO
2 environment. For example, breeding for restricted tillering ability is a current strategy used for dry environments. In the present study we specifically investigated photochemical and photosynthetic responses as well as antioxidative defence capacity of two Triticum aestivum L. cultivars with contrasting tillering ability
(H45, low tillering; Yitpi, high tillering) to the effects of two CO
2
levels (ambient aCO
2
~380 ppm, elevated eCO
2
~550 ppm) in two subsequent years (2009 and 2010).
Plants were grown within the Australian Grains Free Air
Carbon Dioxide Enrichment (AGFACE) facility,
Horsham, Australia. Chlorophyll fluorescence and gas exchange measurements as well as antioxidative defence capacity (ascorbic acid, glutathione) were studied on flag leaves during anthesis. For both years we found that A and Ci were significantly greater in both cultivars when grown under eCO
2
, whereas there was no significant difference in gs. In addition, greater A did not translate into greater phiPSII, there were also no differences in
Fv’Fm’ or qP between CO
2
treatment or cultivars.
Antioxidant analyses are still pending. Our first results suggest that during anthesis flag leaf photosynthesis, although significantly affected by CO
2
, is not different
451

between these two cultivars. In addition, photochemical processes were neither affected by CO
2
treatment nor different between cultivars. The current study is funded by the Australian Commonwealth Department of
Agriculture, Fisheries and Forestry (DAFF) and the
Grains Research and Development Corporation (GRDC).
P0312 – ePoster
Control of sucrose transport and accumulation in sugarcane
Rae, A 1 , Pioch, K 2
Bonnett, G 1
, Grof, C 3 , Casu, R
, Manners, JM
1 , Glassop, D 1 ,
1 CSIRO Plant Industry; of Cologne, Germany; 3
2 Botanical Institute, University
School of Environmental and Life
Sciences, University of Newcastle, Australia
Sugarcane has a highly effective carbohydrate biosynthetic and storage metabolism that facilitates its use for the commercial production of sucrose. Sucrose is stored in the large vacuoles of the parenchyma cells in the culm, where it can reach 60% of the dry weight.
Accumulation of this osmotically active solute relies on partitioning of sugars between tissues, cells and subcellular compartments. As part of a functional genomics approach to increasing sucrose content, EST sequences were isolated and expression analysis was performed to identify genes correlated with sucrose transport and metabolism. By combining these results with functional analysis and localisation, we have developed an integrated model for the pathway of sucrose accumulation. In this pathway, the membrane transport steps are thought to have a large influence on the final sucrose concentration. In particular, the transporters located on the vacuolar membrane have the potential to regulate the ultimate proportion of sucrose retained in the storage compartment. We have identified and characterised several sugar transporters from the sugarcane culm. Recently, two of these transporters were localised to the vacuolar membrane by expression of
GFP fusion proteins. Measurement of radiolabelled sugar uptake by the transporters in a heterologous system defined specificities for glucose and for sucrose respectively, and suggested that both transporters regulate efflux of sugars from the vacuole. Since the final sucrose concentration is the net product of the amount of sucrose stored and the amount remobilised from the vacuole, these transporters are likely to be an important control point for sucrose accumulation.
P0313 – ePoster
In vitro selection of ginger cell lines tolerant to
Pythium myriotylum and its regeneration
Rangappa, K 1 , Masgod Chennaveeraiah, G 2
1 Assistant Professor, Dept of Life Sciences, Kristu
Jayanti College, Bangalore, Karnataka, India;
2 Professor, Dept of Molecular Biology, Bangalore
University, Bangalore, Karnataka, India
Ginger ( Zingiber officinale Rosc.) cell lines tolerant to culture filtrate of Pythium myriotylum were isolated using in vitro selection techniques. The axenic sheathing leaf base explants was cultured on Linsmaier and
452
Skoog’s Basal Medium (LSBM) supplemented with 2,4-
D (9.04 µM) to obtain profuse, compact, creamish white callus. Continuous in vitro selection technique was employed for three cycles to the callus using pure culture filtrate of P. myriotylum to obtain four tolerant cell lines
CL1a1 and CL1a2, CL1b, CL2. The tolerant cell lines were sub-cultured on non-culture filtrate regeneration medium LSBM supplemented with BAP (22.20 µM) and
Kn (9.28 µM) to retrieve complete plantlets. These plantlets were subjected to in vitro sick plot technique and were screened for disease tolerance that revealed the tolerant:susceptible ratio 14:32. The plants retrieved from
CL1a1 and control callus were susceptible under in vitro sick plot selection pressure. The tolerant plants thus obtained were subjected to in vivo sick plot technique that revealed tolerant:susceptible ratio 7:14. The genetic fidelity of the isolated tolerant plants was confirmed through RAPD assay. The axenic disease tolerant clones were hardened and successfully transferred to soil with
62% survival frequency.
P0314 – ePoster
Molecular characterization of bamboo using RAPD markers
Rangappa, K 1 , Srinivasan, B 2
1 Kristu Jayanti College, Bangalore, India; 2 Oxford
College of Science, Bangalore, India
Random amplified polymorphic DNA (RAPD) markers were used to analyze genetic variation among eight varieties of bamboo. Polymerase chain reaction (PCR) with 10 arbitrary decamer oligonucleotide primers applied to eight varieties produced a total of 246 different marker bands of which 4.47% were polymorphic. The 10 primers generated discrete markers with a molecular weight ranging from 0.5 kb to 4.5 kb. Of the 10 primers used, 9 primers generated amplification products and primer OPG 03 did not show amplification in all the varieties studied. The amplified products were used to estimate genetic variability using Dice’s similarity coefficient. The data matrix was converted to a dendrogram, which displayed relative genetic similarities between the varieties. The RAPD analysis offered a rapid and reliable method for evaluation of genetic variability between the varieties, which could be utilized as elegant tool in molecular diagnosis of different varieties of bamboo.
P0316 – ePoster
Study of in vitro callogenesis and organogenesis in
Pimpinella anisum
L.
Raouf Fard, F 1 , Moieni, A 2 , Omidbaigi, R 1
1 Dept of Horticulture, Faculty of Agriculture, Tarbiat
Modares University, Tehran, Iran; 2 Dept of Plant
Breeding,Faculty of Agriculture, Tarbiat Modares
University, Iran
Independent experiments were carried out to investigate the callogenesis potential and indirect shoot regeneration in various genotypes of Pimpinella anisum L. Explants, including stem and root segments, intact cotyledonary

leaves, and shoot tips, were prepared from seedlings grown on MS medium. Root explants from 77 genotypes and shoot explants from 28 genotypes were grown on
MS medium containing 3 mgl
+ 0.5 mgl -1
-1 2,4-D and 2 mgl -1 2,4-D
kinetin respectively. Leaf explants from 50 genotypes and shoot tip explants from 73 genotypes were grown on MS medium containing 3 mgl -1 2,4-D + 0.5 mgl -1 kinetin. The shoot and root explants of every studied genotypes produced callus, which had significant differences in volume. However, in the cotyledonary leaves and shoot tips, only 64.35% and 84.9% of the genotypes produced callus respectively. To study the shoot regeneration, produced callus from shoot and root explants were transferred to MS medium with different hormone treatments. The results indicated that 95.7% of produced callus from shoots had shoot regeneration on
MS medium containing 0.5 mgl -1 BAP, whereas produced callus from roots had the maximum shoot regeneration on MS medium containing 2 mgl -1 BAP and
0.5 mgl -1 NAA.
P0317 – ePoster
Influence of AM fungi on the growth, biochemical and yield parameters of
Lycopersicum esculentum
Mill. (tomato)
Reddy, PG 1 , Andhale, BS 1
1 Padmashri Vikhe Patil College Pravaranagar, India
Investigations on the changes in growth, biochemical and yield parameters of Lycopersicum esculentum Mill.
(tomato), plants inoculated with two arbuscular mycorrhizal fungi viz; Gigaspora margarita and Glomus mosseae were carried out through pot culture experiments. Significant increase in height of the plants inoculated with only G. mosseae was noted. No. of branches, leaf area, inflorescences, flowers, fruits and the content of chlorophyll, nitrogen and phosphorous was more in the plants inoculated with both species. G. margarita alone showed comparatively less effect. The results are compared with control experiments and represented in tabular as well as graphical forms.
P0318 – ePoster
Pathogens and pathogenicity of grain molds of
Sorghum
Reddy, PG 1 , Wabale, HS 1
1 Padmashri Vikhe Patil College Pravaranagar, India
Grain mold is a major disease that develops under moist weather conditions after flowering until grain maturity and before harvest in Sorghum bicolor (L.) Moench. A number of fungal species are responsible for grain mold.
The seeds affected by grain mold show characteristic discoloration and become shriveled. Such seeds lose their market value. Causal organisms decrease the % germination of seeds and increase seedling mortality.
Fungi associated with discolored and shriveled Sorghum grains collected from fields were isolated by standard blotter method and the isolates were cultured on agar medium in petriplates to study the cultural and morphological characters for identification. A total of five fungal pathogens viz; Fusarium moniliforme Sheld.,
Curvularia lunata (Wakker) Boedijn., Alternaria alternata , Phoma sorghina (Sacc.) Boerema.,
Macrophomina phaseolina (Tassi.) Goid. were isolated and their pathogenicity test for assessing the germination inhibition as well as seedling mortality was carried out under in vitro and in vivo conditions by inoculation of spore suspension of each isolate on the surface of grains sterilized with 0.1 % HgCl . The % germination
2 inhibition was maximum (49% in vitro, 45% in vivo) in seeds inoculated with C. lunata and minimum (23% in vitro, 19% in vivo) in those seedlings inoculated with A. alternate , whereas the seedling mortality was high (75% in vitro, 78% in vivo) in seeds inoculated with F. moniliforme and low ( 26% in vitro, 32% in vivo) in those inoculated with M. phaseolina . Overall, the germination inhibition was more in vivo and less in vitro conditions in all cases. The results are supported by observation tables and photographs.
P0319 – ePoster
Effects of fertilizer sources and application rates of
Nitrogen (N) on growth, yield and nutrient status of
Kohlrabi
Saleh, S 1 , El-Bassiony, AM 2 , Fawzy, ZF 2
1 Horticultural Crops Technology Dept, National
Research Centre, Egypt; 2 Vegetable Research Dept,
National Research Centre , Egypt
It is well know the influence of fertilization to achieve a high yield of vegetable crops. Increasing the used of mineral fertilizers causes environmental problems due to the excessive accumulation and leaching of harmful elements to the ground water. Therefore, it is advisable to pay special attention to use safe agriculture system for vegetable production. With the increased environment awareness, organic agricultural production is being more important around the world. The objective of this experiment was to reduce mineral fertilizers in vegetable production without yield loss and to minimize its impact on environment pollution. Two field experiments were carried out in newly reclaimed land at El-Nobaria, northern Egypt during the two successive seasons of
2008/2009 and 2009/2010 to study the response of kohlrabi plants (cv. Grand Duke) to different fertilizer sources and application rates of Nitrogen (N). Five-week old kohlrabi seedlings were transplanted on October 29th and November 1st, respectively into the open field. The experiments were carried out in a split plot design with three replicates. Three fertilizer sources, e.g., mineral N fertilizer (control) as ammonium nitrate (33.5% N), organic N as chicken manure (3.4% N) and combined application of 50% mineral N (ammonium nitrate) + 50% organic N (chicken manure) were assigned to the mainplots, while three N rates, e.g., 60, 80 and 100 kg
N/feddan (4200 m 2 ) were randomized and occupied the sub-plots. Growth characters as plant height, number of leaves per plant, leaf area), chlorophyll content and tuber yield and its characters as well as chemical composition of edible part were evaluated. The important results obtained were as follows:
• Applying of mineral N source ranked the first in increasing kohlrabi yield, followed by the combined application of 50% organic N with 50% mineral N and
453

lastly coming organic N source. Organically fertilized plants resulted in 81% yield containing less nitrate (76%) compared to the mineral N source. The productivity of kohlrabi plants fertilized by 50% mineral N in combination with 50% organic N was not only similar to those fertilized by 100% mineral N, but also had less nitrate content.
• With increasing the application rate of N within the range of 60 up to 100 kg N/fed., all studied plant growth characters, chlorophyll content and tuber yield and its quality increased, but the differences within application rate of 80 and 100 were not great enough to be significant.
• It could be concluded that the economical and useful fertilizer source and application rate of N for the best growth, productivity and tuber quality of kohlrabi plants is combined source of 50% organic N with 50% mineral
N at application rate of 80 kg N/fed. Where there was no great variation obtained within combined source and mineral source and within application rate of 80 and 100 kg N/fed.
P0322 – ePoster
Fluoride discharge from aquifer using
Brassica oleracea based activated carbon
Veeraputhiran, V 1 , Alagumuthu, G 1 , Padmavathy, S 2
1 Chemistry Research Centre, Sri Paramakalyani College,
Tamilnadu, India; 2 Dept of Plant Science and
Biotechnology, Nirmala College For Women, Tamilnadu,
India
Fluoride occurs in some drinking water sources at levels that are hazardous to health. Excess of fluoride (>1.5 mg/L) in drinking water is harmful to the human health.
Various treatment technologies for removing fluoride from ground water have been investigated in the past.
Present investigation of batch adsorption experiments using carbonized form of Brassica oleracea was conducted to remove fluoride from aqueous solution at neutral pH. The influence of contact time, adsorbent dose, adsorbate concentration, temperature and the effect of co-anions, which are commonly present in water has been experimentally verified. The fluoride adsorption isotherm has been modeled by Freundlich, Langmuir,
Redlich-Peterson and Temkin models. Adsorption study obeys Redlich-Peterson isotherm as well as Langmuir isotherms. This adsorbent obeys pseudo second order kinetics for fluoride removal while performing pseudofirst-order, pseudo-second-order, intra particle diffusion and Elovich models. Thermodynaic studies reveal that this adsorption is endothermic in nature. Instrumental analysis XRD, FT-IR, SEM and EDAX gives the conformation about the fluoride binding ability of adsorbent. Field studies also carried out and obtained good success rate for safe, pollution free and eco-friendly drinking water. All the reports from the experiment confirm that Brassica oleracea based adsorbent should improve the drinking water quality by superlative purging of fluoride and fulfill our aim of technical viability, cost-effective and social desirable.
454
P0326 – ePoster
Triticum dicoccoides
, Wild Emmer Wheat: genetic diversity of salinity tolerance
Shavrukov, Y 1 , Langridge, P 1 , Tester, M 1 , Nevo, A 2
1 Australian Centre for Plant Functional Genomics,
University of Adelaide, Australia;
University of Haifa, Israel
2 Institute of Evolution,
Wild Emmer ( Triticum dicoccoides ) is a progenitor of tetraploid wheat and currently grows in environments subject to abiotic stresses, including high salinity. Fiftyfour genotypes originating from nine geographical populations in Israel, and five standard wheats (three durum and two bread wheats) were assessed for salinity tolerance using supported hydroponics. In this study, we summarize two key components that contribute to salinity tolerance: shoot growth in saline conditions relative to control conditions (relative dry weight); and
Na + accumulation in leaves of salinised plants. An additional third component (shoot growth under control conditions) has an indirect role but is important for salinity tolerance in an agricultural context. Variability in these three components was high. Some genotypes showed high overall relative dry weight, having the ability to maintain growth in moderately saline solution, a low-to-moderate Na + accumulation, and high biomass production under control conditions. Genotypes from other populations had very high relative dry weight but grew very slowly, so were of limited agricultural relevance. Six selected genotypes possessing useful qualities for at least one of the tested components of salinity tolerance were re-analyzed, and a genotype from
Gitit in the eastern Samaria steppes was identified as the most promising salt-tolerant line. Further detailed experiments focusing on the wild emmer genotype from
Gitit could unravel the mechanisms of tolerance to salinity in T. dicoccoides . Such salt-tolerant accessions would be of particular use in wheat breeding programs.
P0327 – ePoster
Diversity of aromatic plants and spices in Thailand
Sookchaloem, D
Poopath, M 2
1 , Narongajavana, P 1
, Phutthai, T 2
, Wongprasert, T 2 ,
1 Faculty of Forestry, Kasetsart University, Chatuchak,
Bangkok, Thailand; 2 National Park, wildlife and plant conservation Dept, Chatuchak, Bangkok, Thailand
A study on diversity of aromatic plants and spices in
Thailand was conducted over a three year period to identify the number of species and generate basic information to assist with utilization and conservation.
Basic data were gathered to plan for the survey and collection of specimens. The study was divided into three parts: 1) categorization of aromatic plant diversity; 2) development of a database of the aromatic plants and 3) a chemical study to extract essential oils, concrete and crude extract. In the first part of the research, analysis based on taxonomic studies identified 471 species of 212 genera and 65 families of aromatic plants The highest numbers of species were found in trees (198 species) and climbers (51 species), respectively. The families

Leguminosae, Rubiaceae, Dipterocarpaceae,
Orchidaceae, and Annonaceae had the highest numbers of species. Dominant genera were: Bauhinia,
Caesalpinia, Gardenia, Pavetta, Morinda, Wendlandia,
Shorea, Dipterocarpus, Dendrobium, Aerides, Polyalthia and Goniothalamus . Nevertheless, 91 aromatic plants species were identified as at risk of extinction, with 35 rare species, 44 threatened species and 12 endemic species. There were 130 aromatic species that could be utilized in one or more categories as ornamental (45 species), edible- food (34 species), medicinal (71 species) and miscellaneous purposes.(4 species). In the second part of the study, a database was created for 471 aromatic plants species. The third part, involving the chemical study found that 106 species (85 genera, 42 families) of aromatic plants could provide essential oil.
Furthermore, 88 species (72 genera, 33 families) produced concrete extract and crude extract, using petroleum ether and ethanol, respectively. Of the 88 species, 46 species (39 genera, 25 families) produced concrete extract, while 42 species (36 genera, 22 families) produced crude extract.
P0329 – ePoster
Metabolome analysis of developmental alterations in
Gentian
caused by mineral deficiency
Takahashi, H 1 , Imamura, T 1 , Miyagi, A 2 , Uchimiya, H 2
1
2
Iwate Biotechnology Research Center, Japan;
University of Saitama, Japan
Gentians , herbaceous perennials, are distributed throughout most of the world. In Japan, Gentiana triflora , G. scabra , and their hybrids are cultivated as ornamental flowers because of their attractive flower colors and shapes. However, since most of the key varieties in Japan gentians show strong inbreeding depression, it is difficult to maintain their genetic purity by self-pollination. Recently, explants were cultured clonally to maintain genetic purity, but culture conditions have not been studied systematically, and the essential nutrients of gentian are currently unknown. We therefore investigated the effects of deficiency of the macronutrients potassium (K) and phosphorus (P) in the culture medium. The results showed that the explants could grow under K or P deficiency and P deficiency caused formation of new buds similar to overwintering buds. To elucidate the metabolic mechanism behind the gentian response to mineral deficiency, we performed targeted metabolome analysis using capillary electrophoresis-mass spectrometry. Multivariate analysis using metabolite profiles clearly revealed that characteristic metabolite patterns arise in response to K or P deficiency. Under P deficiency, a severe decrease in energy metabolites was observed, in contrast to K deficiency and the control conditions. P deficiency caused low energy metabolism, which may in turn trigger new overwintering bud formation in vitro. These findings will contribute to understand an appropriate horticultural condition of gentians and triggering bud formation may provide a new strategy for improving explant storage periods and preventing breakdown of parental and hybrid varieties by repeated subculture.
P0337 – ePoster
An effective in vitro propagation of
Jasminum sambac
L. using axillary buds
Wang, R 1 , He, L 2 , Peng, F 3 , Xia, B 4
1 Institute of Botany, Jiangsu Province & Chinese
Academy of Sciences, China
Jasminum sambac L. is one of the popular flowers and economic plants, however, the efficient micropropagation system has not been developed. The purpose of this study is to establish an efficient micropropagation system of J. sambac through in vitro axillary bud culture. The axillary buds sprouting in the basal culture medium WPM were cultured on elongation medium supplemented with 2 mg/L indole-3-butyric
(IBA), 0.5 mg/L benzyladenine (6-BA) and 0.7 mg/L gibberellic acid (GA3). The elongated shoots were cut into nodal segments and then placed on the shoot multiplication media. Maximum number of shoots (11.1 per explants) was induced on medium containing 1 mg/L zeatin (ZT) and 0.5 mg/L IBA. The most effective protocol of rooting (86.4%) was that shoots were first dipped in 450 mg/L naphthalene acetic acid (NAA) solution for 10 minnutes and then subcultured in the basal 1/2 WPM. The plantlets were acclimatized and transplanted to the field successfully with 78% survival rate.
P0340 – ePoster
Selection of thin cell-wall mutants from a barley
TILLING population, for reduced steeping during malt production
1
Washington, JM
Fincher, G 2
1 , Burton, R 2 , Little, A 1 , Stewart, D 3 ,
School of Agriculture, Food and Wine, University of
Adelaide, Waite Campus, Australia; 2 ARC Centre of
Excellence for Plant Cell Walls, School of Agriculture,
Food and Wine, University of Adelaide, Waite Campus,
Australia; 3 Viterra, Adelaide, Australia
The cell walls of the barley starchy endosperm are an impediment to efficient malting, requiring enzymic degradation before hydrolytic enzymes can gain access to the starch and protein substrates contained within the cells. Penetration of water through starchy endosperm cell walls has been identified as a potentially limiting factor in the rate of grain hydration to initiate endosperm modification during steeping and subsequent germination stages. In this project we are investigating the role of cell wall thickness and composition in water movement across the endosperm during steeping, with a view to reducing steep times and hence water use in the malthouse. Barley starchy endosperm cell walls contain approximately 70% (1,3-1,4) beta-glucan. Theoretically, barley with thin cell walls or reduced (1,3-1,4) betaglucan content will allow more efficient water penetration and reduced water usage during steeping. A
Calcofluor-based fluorescence assay has been developed to screen a large population of Flagship TILLING mutants in order to select for thin cell wall mutants. Over
2300 individual half-grains (M3 grain) were crushed and
455

assayed in a 96-well plate format using Calcofluor fluorescence to estimate cell wall content. The remaining embryo-containing portion of grains with reduced cell wall content, were sown to produce M4 grain. A total of
9 plants were identified as showing 25-40% reductions of
(1,3-1,4) beta-glucan content in the bulk grain, compared with the Flagship parent. Individual M4 grains from these plants were re-assayed in the half-grain Calcofluor assay to determine segregation patterns. All of the lines appear to be segregating with up to 20% of grains per line showing a 75% reduction in cell wall material.
Remaining half-grains and whole grains (M5 grain) from these plants will be replanted for further analysis and backcrossing to parent to produce pure grain for multiplication. Pure mutant grain will be used for steeping experiments and malt analysis in order to identify a malting quality barley line that requires less water for steeping.
P0341 – ePoster
Inclusion of a mammalian cGMP response element in the development of a luciferase reporter cGMP responsive promoter system suitable for use in plant protoplasts
Wheeler, JI 1 , Irving, HR 1
1 Monash Institute of Pharmacy Science, Parkville,
Australia cGMP is a second messenger molecule associated with plant responses to hormones, light, nitric oxide and both biotic and abiotic stress. Currently cGMP levels can be assessed using antibody kits designed for mammalian cells or mass spectrometry analysis. Maathuis (2006) used micro-array analysis of Arabidopsis roots to identify genes that are differentially expressed 90 mins after a 30 min exposure to membrane permeable cGMP. Three genes that were up-regulated were NTL1 (At1g33440),
CHX21 (At2g31910) and SOS3 (At5g24270). We have shown that these three genes are also expressed in
Arabidopsis leaves and protoplasts derived from mesophyll cells. Promoters of these genes were cloned into the pLUCTRAP3 vector (Calderron-Villalobos et al.
2006) to generate luciferase fusions. Results of cGMP induced expression of each promoter luciferase clone in freshly isolated Arabidopsis mesophyll protoplasts show slight variations in luciferase expression after cGMP treatment. The NLT1 promoter fragment contains the mammalian cGMP responsive element (RE) identified by
Hum et al (2004). We have inserted three additional cGMP RE into the NTL1 promoter with the aim of enhancing the promoter response to cGMP and report here the effect on luciferase activity in our protoplast system. The development of a cGMP sensitive luciferase reporter system would then allow us to assess receptors such as AtBRI1, AtPSK1 and AtPepR1 with guanyl cyclase activity by measuring differences in luciferase activity. We could also compare the cGMP sensitivity of our enhanced promoter with other enhanced promoters and reporter systems.
P0342 – ePoster
Effects of wood compost for preventing water pollution caused by chemical fertilizer
456
1
Yamamoto, R 1
Grad School of Horticulture In Chiba University, Japan
The quantity of wood waste is increasing in the urban area of Tokyo, and when it is burned, CO
2
gas is emitted which accelerates the greenhouse effect. We have to consider about how to reduce, recycle and reuse these wastes. One of the solutions is to make compost from wood waste, but the use of that compost is not widespread widely. In this study, I focus on its ability to retain nutrients, and consider the possibility of using wood compost to prevent underground water pollution caused by using too much chemical fertilizer on agricultural fields. I prepared some planting pots were filled with soil and compost, some with only soil. Then, liquid fertilizer and water was applied on them. Water leached from the pots was collected. I examined the leached water, and calculated the quantity of NH
4
and
NO
3
in each sample. 3 types of pots were prepared.
'Control' was filled with only soil not any compost,
'Based' was filled from its bottom with compost and its upper portion was filled with soil and 'Mixed' was filled uniformly with a mixture of soil and compost. The control pots were filled with 1000ml soil each, in based and mixed with 900 ml of soil and 100 ml compost each.
Watering and collecting leached water were done five times during each three consecutive days. Liquid fertilizer was added only at the first watering. In result, outflow of NH
4
-N in the first collection are control based mixed. Using compost appears to decrease the flow of NH -N through the soil column and into the
4 catchment basin. There are no definite differences of outflow of NO
3
-N between control and based in all collections, but mixed are less than others clearly. For both NH
4
-N and NO
3
-N leachate concentrations decrease in this order control, based, mixed. Concerning total outflows of chemicals, both based and mixed resulted in smaller outflows of chemicals than control-fertilizer. It can be said that using compost reduces the total quantity of outflow of chemicals by adsorption and immobilization on to compost. By using wood compost on agricultural fields we can prevent some element of water pollution caused by chemical fertilizer. From all of the results, outflows in mixed treatments tend to be less than ones in based treatments. Then we can say that, the function of preventing water pollution is enhanced when compost is mixed into soil rather than used in one place.
P0345 – ePoster
Research advance on cross-breeding between different species of
Taxodium
Yu, CG 1 , Yin, YL 1
1 Institute of Botany£¬Jiangsu Province and Chinese
Academy of Sciences
This paper gives an overview of the advance on the selection and excellent characters of Taxodium crossed
F1 and backcrossed F1. The Taxodium breeding program of the Nanjing Botanical Garden are convinced that controlled Taxodium hybridization promises to combine the best characteristics of superior parents. Clones T.
‘Zhongshanshan302’( T. distichum x T. mucronatum ); T.
‘Zhongshanshan401’ ( T. ascendens x T. mucronatum) ; T.

‘Zhongshanshan118’[( T.
distichum x T. mucronatum ) x
T. mucronatum ], T. ‘Zhongshanshan405’ ( T. mucronatum x T.
distichum ), T. ‘Zhongshanshan406’ ( T. mucronatum x T. distichum ), T. ‘Zhongshanshan407’ ( T. mucronatum x T. distichum ) and T. ‘Zhongshanshan502’ were selected for growth rate and tolerance to alkaline and salt-rich coastal floodplains. They were identified by
SRAP (Sequence-related Amplified Polymorphism) markers. T. ‘Zhongshanshan405’, T.
‘Zhongshanshan406’, and T. ‘Zhongshanshan407’ were selected in 2004. By in 2011, Their mean annual growth of height was 174.4%, 175.2% and 173.8% of the female parent and 131.0%, 131.6% and 130.5% of the male parent, respectively. Their mean annual increment of
DBH was 358.6%, 317.5% and 374.9% of female parent and 174.4%, 154.4% and 182.3% of the male parent, respectively. T. ‘Zhongshanshan502’ were selected in
2005. By in 2011, It mean annual growth of height was
156.6% of the female parent and 118.7% of the male parent, respectively. It mean annual increment of DBH was 289.6% of female parent and 147.3% of the male parent, respectively. All of them have tall and straight tree trunk, and with excellent characteristics, such as fastgrowing, waterlogging tolerance, salt-alkali tolerance, a wide range of adaptability to various habits and landscape effects. They can be planted in Jiangsu,
Zhejiang, Shanghai near Chongqing, Yunnan, etc.
P0347 – ePoster
Antimicrobial activities of essential oil and lemon juice sour on
Staphylococcus aureus
,
Bacillus cereus,
Klebsiella pneumoniae
,
Pseudomonas aerginosa
and
E. coli
strains
Zaheri, A 1
1 Payame Noor University, Iran
Abstract: lemon belongs to the composite family which have a high level of organic acids like citric acid, and because of acid making conditions were used for treatment of many intestinal infections. Aim: this study has been done to evaluate the antibacterial effect of lemon juice and it's essence on pathogen bacteria comparatively. Material and methods: pure essence of the lemon was prepared from Yasuj zarband Co. and the lemon juice was prepared from the south of Iran in two ways: one part of the juice was prepared from the whole fruit of the lemon and the second part of the juice was prepared from the peeled lemon. Anti bacterial effect of different compounds were examined by disc diffusion method on the following strains: S.aureus
(PTCC1112),
E.coli
(PTCC1330), P.aeruginosa (PTCC1074),
K.pneumonia
(PTCC1053), B.cereus
(Clinical). Then
MIC and MBC effect of each compound was tested on strains. Results and conclusion: results showed that the effect of lemon juice is better than essence of lemon.
P0348 – ePoster
Synthesis regulation and key genes expression of triterpenoid from Birch,
Betula platyphylla
Suk.
Yin, J 1 , Zhan, YG 1
1 College of Life Sciences, Northeast Forestry University,
China
One-year-old seedlings of Betula platyphylla was used as experimental material. The effects of light, moisture, nitrogen nutrition on synthesis, the distribution and variation pattern of temporal and spatial of triterpenoid in the growth process of white birch saplings were studied.
Tissue-specific expression of key enzyme genes of triterpenoids was analyzed by fluorescence quantitative
PCR method. The aim of the study was therefore to explore the mechanism of efficient synthesis of terpenoids, and to lay the foundation for metabolic regulation of triterpenoid from birch. The main conclusions are as follows: (1) One-year-old birch seedling could synthesize betulin and oleanolic acid triterpenoid. Triterpene substance mainly accumulated in bark. The content was very low in root, skin and leaf, even which could not be detected. The accumulation of two kinds of components in the various parts of plant was significantly affected by season and environment factors. During the growing season (from May to
October), the accumulation peak of secondary metabolites in stem bark was from the end of July to late
August. Low concentration of nitrogen could promote accumulation of oleanolic acid in the root, skin and leaf, while high concentration of nitrogen was more beneficial to the accumulation of oleanolic acid and betulin. The triterpenoid accumulation in bark was effectively promoted after MeJA spraying. The highest content treated by MeJA(100uM/L) was 9.370mg/g (2 times of control) at the end of July. Medium (water 35%), and mild drought stress (water 50%), promoted accumulation of the oleanolic acid and betulin in stem bark, respectively, while severe deficiencies of water (water
15%) inhibited synthesis of triterpenoids. Red, yellow, blue or green light resulted in lower accumulation of triterpenoid in stem bark than control (white light).
Among these treatments, green and blue light were more effevtive than others. Proper shading (50% natural light) was also beneficial to the synthesis of triterpenoid. (2)
During the growing season, the gene expression level of
BPW and BPY was the lowest in leaves, while the highest in stem cambium from end of July to end of
September. The expression of BPY gene was high in root from end of July to end of October, the BPW gene showed the highest expression level in root in August, and the highest expression level of BPX arose at the end of August, but its product was comparatively low. The mRNA level of triterpenoid related genes (BPW, BPX and BPY) took similar accumulation and distribution to triterpenoid constituents, and both them were markedly regulated by the season.
P0350 – ePoster
Evaluation of genetic diversity of pecan cultivated in
China using SSR and ISSR markers
, Tao, W 1 , Jiping, X 1 , Fei, Zhongren, G
X 1
1 , Xiaodong, J 1
, Yuhong, Z 1
1 Jiangsu Institute of Botany, Chinese Academic of
Sciences, Nanjing, PR China
Pecan ( Carya illinoensis (Wangenh.) K.Koch) has been introduced into China for over 100 years. Now, there are
457

hundreds of varieties, cultivars in China, including cultivars introduced from American, domestic cultivars and cultivars selected from seedling pecan. In this study, simple sequence repeat (SSR) and inter-simple sequence repeat (ISSR) markers were utilized to reveal the genetic diversity of pecan grown in China. 81 samples, including
18 introduced cultivars, 10 domestic cultivars, 53 pecan seedlings with good qualities, together with Carya cathayensis Sarq. and Juglans nigra L. were analyzed. A total of 8 informative SSR primer pairs and 10 ISSR primers were chosen following an initial screening of 19
SSR primer pairs and 46 ISSR primers using 3 DNA samples. 68 and 94 DNA bands were detected, of which all the bands were polymorphic. SSR results showed that the effective number of alleles (Ne) was 1.4651, Nei’s gene diversity (H) was 0.2858 and Shannon’s information index (I) was 0.4409. This information indicated that the main genetic variance existed among cultivars. Genetic variance analysis showed the coefficient of gene differentiation among cultivars (Gst) was 0.9661 and the gene flow among cultivars (Nm) was
0.0175, which means 96.61% differences were existed among cultivars. Genetic distances, based on the similarity coefficient of Nei & Li, varied from 0.96 to
0.38, with an average of 0.72 among all cultivars. ISSR results showed that the effective number of alleles (Ne) was 1.4534, Nei’s gene diversity (H) was 0.2732 and
Shannon’s information index (I) was 0.4216. Genetic variance analysis showed the coefficient of gene differentiation among cultivars (Gst) was 0.9803 and the gene flow among population is 0.0101. Genetic distances, based on the similarity coefficient of Nei & Li, varied from 1.00 to 0.50, with an average of 0.74 among all cultivars. The Gst based on ISSR and SSR were
0.9661 and 0.9803 and very close, which indicated that the pecan cultivars had significant diversity in DNA level. The Nm was very small may result from the selfpollination. All this data suggested that there were genetic differentiation among cultivars. Though they were cultivated for many years, they kept rich genetic diversity among cultivars. These results provided molecular data for the genetic improvement and breeding of pecan.
P0351 – Poster
Phytotoxic effects of six surfactants on
Allium cepa
L. plantlets
Akbas, H 1
S 1
, Dane, F 1 , Yilmaz, G 1 , Ozturk, F 1 , Leventer,
1 Trakya University, Edirne, Turkey
Surfactants are used to improve the effectiveness of foliar applied herbicides, other pesticides and defoliants by reducing the surface tension of aqueous systems.
However, there is evidence that surfactants are capable of exerting inhibitory as well as stimulatory effects upon plant growth and behavior. The phytotoxic effects of two anionic surfactant, Sodium dodecyl benzene sulfonate
(DBSNa) and Sodium dodecyl sulfonate (SDS), one meric and one dimeric cationic surfactant,
Cetyltrimethylammonium bromide (CTAB) and Gemini surfactant (16-2-16) and two non-ionic surfactant
Polyoxyethylene octyl phenyl ether (Triton X-100) and
Polyoxyethylene 23 lauryl ether (Brij 35) were evaluated
458 under laboratory conditions using onion ( Allium cepa L.) as a test material. The phytotoxic effects and the change of pH in solutions were observed. The Phytotoxic effect on root were determined after 24, 48, 72, 96, 120 and 144 hr of treatment under continuous light at 16±1°C.
Surfactants used in the study were tested at 0.5%, 0.2%, and 0.1% (w/v) concentrations. Some of the phytotoxic effects of anionic and cationic surfactants were observed in onion. Also, non-ionic surfactants have shown some stimulatory effects. Stimulatory effects occurred at all non-ionic surfactant concentrations and these effects were more Brij 35 than Triton X-100. Also, stimulatory effects were observed at more high level at the low
Triton X-100 concentration than at the high. However, some differences in effects were observed between the six types of surfactant. The pH values were decreased with time at all surfactant concentrations.
P0352 – ePoster
Development and utilization of the Yellow Lotus germplasm in Thailand
, La-ongsri, W 2 , Boonsirichai, K 1
1
Puripunyavanich, V 1
Thailand Institute of Nuclear Technology (Public
Organization), Nakhon Nayok, Thailand; 2 Botanical
Garden Organization, Chiang Mai, Thailand
Being in a tropical climate, Thailand hosts only a single species of lotus: Nelumbo nucifera (Gaertn.), represented by six varieties. These plants offer only white or red flowers. In temperate regions, such as southern United
States and China, Nelumbo lutea (Willd.) thrives and gives yellow flowers in June through July. Many failed attempts have been made to raise N. lutea to flowering in
Thailand in hopes of transferring the yellow-flower trait into local N. nucifera . We resorted to radiation-induced mutagenesis. Rhizomes were treated with 10Gy gamma radiation and vegetatively propagated. A mutant was identified in the second generation, which produced flowers. It appeared self-sterile but could be vegetatively propagated and still produced flowers. The mutant flowers produced yellow petals pistils and stamens, similarly to wild-type N. lutea . However, the mutant petal shape is oblong instead of obovate. Its pistil contains only one to three ovaries stead of multiple ovaries. The mutant enters a dormant state during the cool period of the year in December through February and flowered only during the long-day period in June through August. Vegetative propagation is possible but attempts are being made for in vitro propagation to mass produce the mutant. Despite being self-sterile, the mutant pollens are active and were used successfully to pollinate local N. nucifera in order to transfer the yellow-flower trait to local varieties. Two hybrids were constructed, namely Chandrakomen and Yellow Chutra.
Chandrakomen is a hybrid between a red lotus and the mutant yellow lotus ( N. nucifera cv. Bua Luang
Phrarachinee x mutant N. lutea ). Its flowers produce red and yellow petals, with the red color receding from the petal base toward the petal tips over the blooming days similarly to Mrs.Perry D. Slocum which is also a hybrid between a red N. nucifera and the American N. lutea . On the last blooming day, its petals appear creamy yellow.
Attempts to self-pollinate Chandrakomen have not been successful. On the other hand, Yellow Chutra came from

a cross between a white-flower lotus and the mutant N. lutea of ( N. nucifera cv. Sattabut x mutant N.
lutea ) and produces whitish yellow petals.
P0353 – ePoster
A weakly photoperiod sensitive mutant of Khao Dawk
Mali 105 and its tissue culture method
Boonsirichai, K 1
Taprab, S 3
, Phadvibulya, V 1
, Puripunyavanich, V 1
, Klakhaeng, K 2
, Chookaew, S 1
,
,
Khamvarn, V 1 , Sungsinleart, K 1
1 Thailand Institute of Nuclear Technology (Public
2
Organization), Nakhon Nayok, Thailand;
Chachoenghsao Rice Research Center, Chachoengsao,
Thailand; 3 Rice Research and Development Bureau, Rice
Dept, Bangkok, Thailand
Khao Dawk Mali 105 (KDML105) is widely known in world markets as Thai jasmine rice. It is a photoperiodsensitive, short-day cultivar and only yields a crop a year, being planted in July and harvested in November.
Because of its high export value, we isolated a weakly photoperiod-sensitive mutant of KDML105, which can flower all year round allowing three crops per year in irrigated regions. KDML105 seeds were bombarded with
20 Gy fast neutron particles. Five mutant sibling lines including 20-200-1, 20-200-4, 20-200-8, 20-200-16 and
20-200-19, which were derived from a single parent, were selected for further propagation. These mutant lines showed an early-flowering phenotype with similar plant morphology to KDML105. However, 20-200-16 later gave a progeny with short stature, which we named 20-
200-16NPT. The degree of photoperiod sensitivity of 20-
200-4 placed the mutant into the weakly photoperiod responsive group. The mutant appeared recessive. An F2 mapping population was generated between 20-200-4 and Hangyi 71, a photoperiod-sensitive cultivar. Seeded in April, the F2 progeny was later categorized into two groups: one which flowered within 120 days after seeding and one which flowered after 120 days after seeding, on which bulk segregation analyses were performed. Two DNA bands, which correlated to the photoperiod-insensitive phenotype (flowering during the long-day period), were identified through AFLP fingerprinting with primer pairs EcoRI-AGC/MseI-GCA and EcoRI-ATC/MseI-CAC. Further studies are still needed to identify a linked DNA sequence. On the hand, tissue culture methods were being developed for
KDML105 and 20-200-16NPT in order to provide a platform for future radiation-induced mutagenesis using irradiated calluses. Calluses were first obtained from embryo being cultured on medium containing 2 mg/l 2,4-
D with or without 1g/l L-proline and 3mg/l L-lysine.
KDML105 yielded comparable percentages of callusforming embryos on all media, ranging from 76.98 –
82.15%. 20-200-16NPT gave the best callus yield on medium without L-proline and L-lysine with 71.36% callus-forming embryos, while the smallest yield of
23.88% callus-forming embryos was obtained when cultured on medium with both L-lysine and L-proline.
These calluses will be further tested for their ability to develop green shoots on shoot induction media containing varying amounts of auxin and cytokinin in the future.
P0354 – Poster
Histological and cytological observations on early development phases of calli in cultures of anthers of
Oryza sativa cv. Baldo
Dane, F 1 , Meric, C 1
1 Trakya University, Erdine, Turkey
Microspores have the remarkable capacity to develop into haploid plants via embryogenesis in vitro. In addition, microspore embryogenesis can be used to study plant embryogenesis and phase transitions during the alternation of generations in plants. We carried out a systematical study on the histological and cytological during different development phases of calli. Anthers containing uninucleate microspores from Oryza sativa cv. Baldo were cultured on N6 medium and evaluated for callus induction. Results showed that the callus developed nodule-like structures, which were formed by small, tightly packed, hemispherical cells. Calli at different development stages were fixed for histological and cytological examinations, cut 10 µm thick sections, stained with PAS and IKI and examined using light microscopy. The cytological and histological examination showed that the cells of calli had similar morphological characteristics. It was observed that cells had vacuolation of cytoplasm and amyloplasts. This paper describes callus formation from cultured anthers with uni-nucleate microspore of rice and examines cytologic and histologic properties of callus, and also aims to understand the mechanisms of callus morphogenesis of rice.
P0355 – Poster
Contrasting hydraulic regulation in closely related forage grasses: implications to breeding for drought resistance
Holloway-Phillips, MM 1 , Brodribb, T 2
1 Tasmanian Institute of Agricultural Research,
University of Tasmania, Australia; 2 School of Plant
Science, University of Tasmania, Australia
Plant traits that improve crop water use efficiency are highly sought after but difficult to isolate. Here we examine the integrated function of xylem and stomata in closely related forage grasses to determine whether quantitative differences in water transport properties could be used to predict plant performance under waterlimited conditions. Cultivars of two forage grass species with different drought tolerance ratings, Lolium multiflorum Lam. and Festuca arundinacea Schreb., were assessed for maximum hydraulic conductivity
(Kmax), vulnerability of xylem to hydraulic dysfunction
(P50) and stomatal sensitivity to leaf water potential.
Species-specific differences were observed in several of these traits, and their effect on whole-plant performance was examined under well-watered and restricted watering conditions. It was shown that whilst P50 was comparable between species, for F.
arundinacea cultivars there was greater hydraulic risk associated with reduced stomatal sensitivity to leaf hydration. In contrast, L. multiflorum cultivars expressed a higher capacity for water transport,
459

but more conservative stomatal regulation. Despite different susceptibilities to leaf damage observed during acute drought, under the sustained moderate drought treatment, the two strategies were balanced in terms of water conservation and hydraulic utilisation resulting in similar dry-matter production. Characterisation of water use patterns according to the key hydraulic parameters is discussed in terms of implications to yield across different environmental scenarios as well as the applicability of water transport related traits to breeding programs.
P0356 – Poster
Nuclear DNA contents of some
Ophrys
L. species
(Orchidaceae) growing in European Turkey
Meric, C 1 , Guler, N 1 , Arda, H 1
1 Trakya University, Erdine, Turkey
In this study, nuclear DNA contents of eight taxa belonging to the genus Ophrys ( Ophrys apifera , O. fusca ,
O. lutea subsp.
minor , O. mammosa , O. oestrifera subsp. oestrifera , O. oestrifera subsp. heldreichii , O. umbilicata , O. vernixia subsp. vernixia ) naturally growing in Trakya region (European Turkey) were analyzed for the first time by flow cytometry. All investigated species were found to be diploid. The 2C
DNA amounts were determined as 19.35 (±0.25) pg for
O. apifera , 20.74 (±0.11) pg for O. fusca , 19.78 (±0.36) pg for O. lutea subsp. minor , 21.24 (±0.18) pg for O. mammosa , 21.34 (±0.30) pg for O. oestrifera subsp. oestrifera , 20.12 (±0.60) pg for O. oestrifera subsp. heldreichii , 18.58 (±0.20) pg for O. umbilicata and 18.85
(±0.42) pg for O. vernixia subsp. vernixia ). This study provides additional data on nuclear DNA content of
Orchidaceae taxa.
P0357 – Poster
Volatile components of a nocturnal flower,
Anemarrhena asphodeloides
Bunge (Agavaceae)
Murata, H
A 1
1 , IWABUCHI, H
, MURATA, J 3
2 , INATOMI, Y 1 , INADA,
1 Faculty of Pharmaceutical Sciences, Setsunan
University, Japan;
USA; 3
2 San-Ei Gen F.F.I., Inc., New York,
Botanical Gardens, University of Tokyo, Japan
The rhizome of Anemarrhena asphodeloides , native of northeast China, is a popular Chinese medicine 'Zhi Mu'.
It is introduced to Japan and sometimes grown in the herbal gardens but the flowering behavior may not be well known. The flowers are closed during the day then fully open in the evening and very fragrant through the night. Small moths were observed to visit at night. The fragrant flowers were collected at night, extracted with hexane and the extract including endogenous volatile components was analyzed using gas chromatographymass spectrometry. Various components including the esters of aromatic and fatty acidic series, benzoic acids and phenylacetic acids were recognized. It is notable that a unique gamma-nonalactone with coconut-like sweet smell as well as methyl cinnamate and 3-hydroxy-4phenylbutan-2-one were also recognized.
460
P0359 – Poster
An investigation of the effects of nonionic surfactant with herbicide on
Triticum aestivum
L. by flow cytometry
Yilmaz, G 1 , Dane, F 1 , Meric, C 1
1 Trakya University, Erdine, Turkey
The aim of this study is to show the effects of non-ionic surfactant (alkylethersulfate sodium salt) with herbicide
(propoxycarbazone-sodium) on the wheat ( Triticum aestivum L.) using flow cytometry. It is a technique for counting and examining microscopic particles, such as cells and chromosomes, by suspending them in a stream of fluid and passing them by an electronic detection apparatus. Three different doses of (0.42 mM, 0.84 mM,
1.68 mM) propoxycarbazone-sodium and the same doses with 0.25 % (alkylethersulphate sodium salt) were applied to Triticum aestivum L. as seed treatment. After seed treatment, for flow cytometric analysis, the young leaves of growing plants were prepared. Fresh leaf fragments of samples were chopped with a razor blade on ice in a plastic petri dish containing MgSO buffer,
4 dithiothritol, propidium iodide and triton X- 100. Nuclei, isolated from leaves of wheat were stained with propidium iodide (PI) and found suitable for DNA analysis by flow cytometry. The prepared materials were analysed by EPICS XL model flow cytometer (Beckman
Coulter). Analysis were performed three times, averages and standart deviations of measures were taken. The mean DNA content per sample plant was based on 10000 scanned nuclei. DNA amounts were given by flow cytometry graphics of increasing doses of
(propoxycarbazone-sodium) with and without
(alkylethersulphate sodium salt). When nuclear DNA contents in leaves were compared with control group,
DNA contents increased by increasing doses of
(propoxycarbazone-sodium) with non-ionic surfactant.
STATISTICA AXA 7.1 statistical program which has number AXA507C775506FAN3 was used for statistical analysis.
P0361 – ePoster
Spatiotemporal gene expression profiling of
Oryza sativa
ssp. japonica
cv. Nipponbare
Antonio, B 1
Kamatsuki, K
,
2
Sato, Y 1 , Takehisa, H
, Sugimoto, K 1
1 , Namiki, N
, Nagamura, Y 1
2 ,
1 National Institute of Agrobiological Sciences, Tsukuba,
Japan; 2 Mitsubishi Space Software Co. Ltd, Tsukuba,
Japan
Understanding the function and regulation of all the rice genes essential for vegetative and reproductive growth is one of the major goals of cereal genetics to ensure the

development of improved varieties. To address this goal, we performed spatiotemporal gene expression profiling of the japonica rice cultivar Nipponbare using 48 different tissue and organ types representing the entire growth and developmental cycle from transplanting to harvesting under natural field conditions. Transcriptome analysis was performed with the Agilent 44K rice microarray platform, which contains 35,760 independent probes corresponding to 27,201 annotated loci in the rice genome. We obtained a total of 143 microarray data representing triplicate expression profiles for each organ and/or tissue sample. The number of expressed genes across organs/tissues did not vary significantly and ranged from 63-76% with about 43% (15,224) of the transcripts were expressed in all organs/tissues. Principal component analysis revealed three distinct transcriptome clusters corresponding to the profiles of vegetative organs such as leaf, stem, and root; reproductive organs such as anther, pistil, and entire inflorescence; and the endosperm. Based on the degree of specificity for a particular organ or tissue as estimated by the Shannon entropy scores, a total of 731 organ/tissue-specific genes corresponding to 660 loci were identified including leaf-, root- and seed-specific gene clusters, and genes specifically expressed in floral organs which were mostly found in anther. In the leaf blade, leaf sheath, and root, the expression of many genes also showed growth-stage specific signatures. Additionally, we extracted 215 genes that universally showed changes in expression in these tissues from vegetative to reproductive stages. Analysis of the transcriptomes of vegetative organs at daytime and nighttime also showed a diurnal pattern in gene expression which is particularly prominent in the leaf. A global gene expression profile of the rice plant during the entire life cycle provides an opportunity for clarifying the expression of many genes that are likely to regulate major developmental changes and govern the manifestation of agronomically important traits, as well as in deciphering the most probable function of many genes that until now remain uncharacterized. All the data generated from this spatiotemporal gene expression profiling can be accessed via our database, RiceXPro
(http://ricexpro.dna.affrc.go.jp/). With the expression profile of all genes that comprise the genome, we hope to provide reference information that can be used not only in functional characterization of all genes in rice but also in elucidating the global transcriptional changes encompassing the entire growth under natural field conditions.
P0362 – ePoster
QTL mapping of trichome density in two new RIL populations of
Arabidopsis thaliana reveals environment-specific and herbivory-response loci
Bloomer, R 1 , Symonds, VV 1
1 Massey University, New Zealand
Leaf trichome density, a highly variable and selectively important trait in natural populations of Arabidopsis thaliana , is an interesting model system for understanding the genetic basis of natural phenotypic variation. In A. thaliana leaf trichomes are single-celled and branched, serving to protect the plant from insect herbivory. Quantitative trait locus (QTL) mapping can provide insights into the genetic architecture underlying trichome density variation in natural populations, but those insights are limited by the amount of genetic variation and number of experimental environments sampled. Here, we present the mapping results for two previously uncharacterised A. thaliana recombinant inbred line (RIL) mapping populations, St-0 x Sf-2 and
Hi-0 x Ob-0: trichome density, in both control and simulated herbivory environments. These populations will ultimately be genotyped with approximately 70 markers each, with at least 30 markers in common to facilitate linkage map alignments. Although trichome density has been mapped previously, mapping in new populations and in multiple environments captures additional natural genetic variation and will potentially identify more loci of interest. The natural accessions St-0 and Sf-2 have very similar trichome densities while Hi-0 and Ob-0 have relatively different densities, providing an interesting contrast. QTL were mapped for trichome density in control and simulated herbivory environments in both populations; in addition, the difference in trichome densities between control and simulated herbivory environments was calculated for each RIL as a measure of the plants’ response to damage. High heritability and evidence of transgressive segregation was found in both environments and in response to damage in both populations. Early mapping results indicate that some QTLs appear in both environments, while others are unique to one environment or to response. Some
QTLs map near physical positions of known epidermal development genes, suggesting possible candidate genes for further analysis.
P0363 – ePoster
The impact of variation in regulatory genes on
1 anthocyanin production in
Arabidopsis thaliana
Butcher, M 1 , Symonds, V 1
IMBS, Palmerston North, New Zealand
Anthocyanins are a group of phenolic pigments unique to plants produced via a specialised flavonoid pathway under the regulation of a complex network of transcription factors. Anthocyanins act as a generic stress response mechanism to both internal and external stimuli in many plant species due to their various phytoprotectant properties. As such, natural variation observed in anthocyanin production may have important consequences for acclimation at an individual level and adaptation at the population level. Using association mapping in a core population of 96 ecotypes, we are investigating the genetic architecture of anthocyanin production in the model species, Arabidopsis thaliana .
Mapping results from two environments will be presented, along with candidate gene data. Our findings will contribute both to understanding the roles and regulation of anthocyanins as well as the rapidly expanding story of development throughout the life of a plant.
P0364 – ePoster
Genetic diversity and population structure of a broad range of olive genotypes assessed by SSR markers
461

Borges do Val, AD 1
Neto, J 3
, Ferreira, JL 2 , Pasqual, M 1
, Soares, BDF 2 , Cancado, GMA 2
, Vieira-
1 Dept of Agriculture, Federal University of Lavras
2
(UFLA), Lavras, MG, Brazil
Plant Biotechnology Laboratory, Agricultural Research
Agency of Minas Gerais (EPAMIG), Caldas, MG, Brazil;
3 Ituporanga Experiment Station, Santa Catarina State
Agricultural Research and Rural Extension Agency
(EPAGRI), Ituporanga, SC, Brazil
Olive ( Olea europaea L.) is an important crop worldwide and Brazil has recently begun to produce olive oil at commercial scale. EPAMIG, an agricultural research agency of Minas Gerais State, was responsible for the initial establishment of this crop in Brazil and it still remains as the major research and technology developer for this specie in the Brazilian environment. It is crucial for the correct management of the olive breeding program to know in detail the genetic diversity and population structure of the olive germplasm available in
Brazil. Thus, the aim of this study was the DNA fingerprinting and characterization of genetic interrelationships among sixty olive genotypes belonging to
EPAMIG’s germplasm by using SSR markers. The genotypes sampled included Brazilian genetic accesses and most of commercial varieties cultivated worldwide.
We used 12 SSR markers specific for olive: GAPU101,
GAPU11-17, UDO99-009, UDO99-019, GAPU45,
GAPU71A, GAPU71B, GAPU89, GAPU59, UDO99-
031, GAPU12, and UDO99-039. Statistic analyses were performed with the Convert, GeneAlex v. 6 (Matching,
Principal Coordinate Analysis – PCoA, and Probability of Identity), Powermaker v. 3.25 (Summary Statistics and Neighbor Joining Tree), and Structure v. 2.3.3 applicative. The average number of alleles identified was
6: GAPU101 generated 10 and GAPU71B generated 9 while GAPU45 generated only 3 alleles. The highest
Gene Diversity and Polymorphism Information Content were found in GAPU101 (0.8399 and 0.8203) and
GAPU71B (0.8117 and 0.7863) while the marker
UDO99-019 presented the lowest levels (0.1976 and
0.1860). The multilocus matching identified 52 unique profiles out of 60 accessions. The high sensitivity of SSR markers allowed us to identify some synonyms and homonyms among the varieties evaluated. Furthermore, the 12 SSR were able to provide a probability identity of
1.52 x 10-10. The first two components of PCoA explain
48.16% of the genetic variation and the 60 accessions structured in 4 major groups. Eighteen hybrids (30%) were detected considering the 80% cut-off of membership coefficient. The congruence between
Bayesian Structure and the Unrooted Neighbor Joining
Tree methodology was very high indicating strong consistency between the results obtained. In order to explore the genetic variability available in the olive germplasm, these results will be useful to guide breeders during the best parental choice. The authors would like to acknowledge financial support from FAPEMIG, IFS,
PNPD-CAPES, EMBRAPA, FINEP, and CNPq.
P0365 – ePoster
Comparison of a retrotransposon-based marker with microsatellite markers in the genus
Vitis
San'Ana, GC 1 , Ferreira, JL 2
Soares, BDF 2
, Souza, RAV
, Cançado, GMA 2
3 , Borem, A 1 ,
1 Dept of Genetics, Federal University of Viçosa (UFV),
Viçosa, MG, Brazil; 2 Plant Biotechnology Laboratory,
Agricultural Research Agency of Minas Gerais
(EPAMIG), Caldas, MG, Brazil; 3 Dept of Agriculture,
Federal University of Mato Grosso (UFMT), Cuiabá,
MT, Brazil
The grapevine is a perennial fruit of greater economic importance worldwide. Brazil, although not among the world's largest producers, has been showing growing development in the wine and juice industry, and also has increased markedly your production with the planting of new areas and renovation of vineyards. The correct identification and knowledge of genetic diversity are fundamental information for reasonable management and use of grapevine germplasm. Recently, new types of molecular markers have been developed, such as retrotransposon-based markers. Because of its multilocus pattern, this kind of molecular marker might be able to differentiate several grapevine accessions with just one pair of primers. In order to evaluate the efficiency of this type of marker, we did a comparison between the retrotransposon marker TVV1 against 7 microsatellite markers frequently used in the genus Vitis (VVMD7,
VVMD25, VVMD5, VVMD27, VVMD31, VVS2, and
VZag62). The reference population used in this work consisted of 26 accessions of Vitis , comprising 7 Vitis vinifera varieties, 4 North American varieties and 15 rootstock hybrids. Individually, the TVV1 and the 7 microsatellite markers were capable to discriminate all accession except the ‘Niagára branca’ from ‘Niagára rosada’. A point mutation may clarify why these two kind of molecular markers were not able to distinguish those two accessions. Furthermore, using the software
Structure, the retrotransposon marker TVV1 generated 2 clusters: One with V. vinifera plus North American varieties and another only with the rootstock hybrids. On the other hand, the 7 microsatellite markers evaluated together were able to generate 5 clusters: One only for V. vinifera , another only for North American varieties, while the rootstocks hybrids were distributed among the three remaining clusters. The percentage of variation explained by the first two components in the Principal
Coordinate Analysis were 65.21 (TVV1) and 50.42
(microsatellite markers). The Mantel correlation between the distances matrixes generated by both markers was
42.5%. These results evidenced the usefulness of TVV1 marker in DNA fingerprinting, but they also showed the lack of efficiency of TVV1 during the discrimination of structured groups. The authors would like to thank the financial support from IFS, FAPEMIG, FINEP,
EMBRAPA, CAPES and CNPq.
P0366 – ePoster
Genetic variability of strawberry intervarietal hybrids determined by inter simple sequence repeat markers
Nunes, CF
MSC 1
1 , Fernandes, MCN
, Ferreira, JL 1
Cancado, GMDA 1
1 , Ribeiro, AP 2 , Dias,
, Fontes-Soares, BD 1 , Juhász, AC 1 ,
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1 Agricultural Research Agency of Minas Gerais
(EPAMIG), Brazil;
(UFLA), Brazil
2 Federal University of Lavras
Strawberry is an important agricultural crop in Brazil.
However, most of the commercial genotypes currently cultivated there were developed in other countries with environment adaptation often inadequate for the
Brazilian condition. In this work, the Inter Simple
Sequence Repeat (ISSR) technique was used to access the genetic variability and the loci segregation profile of eighty four individual strawberry hybrids obtained from a breeding program developed by the Agricultural
Research Agency of Minas Gerais State (EPAMIG). The aim of this program is to develop new strawberries cultivars with reduced susceptibility to diseases and pests that affects this specie in Brazil as well as improving the fruit quality for industry and fresh consumption. The hybrids were obtained from the crossing between
[Toyonoka x Sweet Charlie], [Camino Real x Sweet
Charlie], [Oso Grande x Sweet Charlie], [Oso Grande x
Toyonoka], [Dover x Oso Grande], and [Camino Real x
Toyonoka] and fourteen genotypes for each hybrid combination were evaluated. The ISSR primers used to screening strawberries genotypes were
5´VBVACACACACACACAC3´,
5´BDBCACACACACACACA3´, 5´HBH
CTCTCTCTCTCTCT3´, 5´GCVTCTCTCTCTCTCTC3´ and 5´BDV AGAGAGAGAGAGAG3´ where V is non
T, B is non A, H is non G, and D is non C. Although the octaploid nature of the strawberry genome complicates interpretation of molecular data, since each locus can hold up to eight different alleles, the ISSR markers might be the most adequate choice for genotyping these genotypes due its low cost, easy handling, high reproducibility and stability associated with its great power of genome sampling. The preliminary results obtained so far show that the genetic profile among the hybrids genotypes into each crossing is very diverse mainly due the high heterozygosis of the genome constitution from each progenitor involved. These results indicate there is a broad genetic background in these genotypes that might allow breeders to obtain new cultivars with adequate agronomic traits for this specie.
However new genepools sources, mainly for resistance and tolerance to biotic and abiotic stress, must be incorporated to achieve success in the breeding program of strawberries in Brazil. The authors of this work would like to thank the financial support from FAPEMIG, IFS,
PNPD-CAPES, EMBRAPA, FINEP, and CNPq.
P0367 – ePoster
Outcrossing rate in olive (
Olea europaea
L.) by microsatellite and ISSR molecular markers
Souza, RAV
Borem, A
GMDA 2
3
1 , Braga, FT 2
, Pasqual, M 4
, Ferreira, JL 2 , Azevedo, PH 1
, Vieira-Neto, J 5
,
, Cancado,
1 Federal University of Mato Grosso (UFMT), Brazil;
EPAMIG, Brazil;
4
3
2
Federal University of Viçosa (UFV),
5
Brazil; Federal University of Lavras (UFLA) , Brazil;
EPAGRI, Brazil
Studies of cross-pollination are very important to understand the evolution of species and to drive their improvement. Olive is cited in the literature as crosspollinating plant, but no one knows the magnitude of this mating system. The aim of this study was to determine the rate of cross-pollination in olive progenies grown in vitro, using both microsatellite (SSR) and ISSR markers.
Young leaves and ripe fruits were collected from the genotypes ‘Ascolano USA’ and ‘Grappolo 541’ in the olive germplasm bank located at the Experimental Farm of EPAMIG in Maria da Fé, Brazil. The plant DNAs were extracted immediately after collection of leaves in the field and also in the progeny when they reached five to six leaves in vitro. Twenty three seedlings per each genotype were randomly chosen for DNA extraction and
10 ISSR and 2 microsatellite markers were used to evaluate the genetic segregation. We analyzed the rate of outcrossing (multilocus tm, single locus outcrossing ts), the paternity correlation (rp), the fixation index Wright multilocus (Fm) using the software MLTR and the progeny heterozygosity through the applicative
POPGENE version 32. The ISSR markers scrutinized 84 and 92 loci for the progeny of ‘Ascolano USA’ and
‘Grappolo 541’ respectively, while the SSR markers together generated 7 and 8 alleles for ‘Grappolo 541’ and
‘Ascolano USA’ respectively. Regardless of marker type used, the set of parameters (individually and collectively) produced enough support to the hypothesis of allogamy in olive as showed by the rate of outcrossing congruent close to 1.00 as well as due the low coefficient of inbreeding (Fm = -0200). The authors of this work would like to thank the financial support from EMBRAPA,
FAPEMIG, FINEP and CAPES.
P0368 – ePoster
1
Horizontal gene transfer between flowering plants
Chen, Q 1 , Chen, GL 1
Inner Mongolia University, Huhhot, P.R.China
Horizontal gene transfer (HGT), the transfer of genes between non-mating species, plays a major role in flowering parasitic plants' genomic evolution. Direct host-to-parasite contact, which includes the manner that different parasites co-attack the same host individual and generalist parasites' attack, has emerged as a common mechanism of HGT. Both the host and the parasite could serve as a vector of HGT between two unrelated species, which involves parasitic plants as donors or as recipients.
Most cases of HGT uncovered thus far have involved genes in mitochondria genome, and recent work demonstrated that nearly identical nuclear genes are present in two far related species, which are facilitated by the active DNA uptake system possessed by the mitochondria and the nuclear. Genes in other organelles are not prone to HGT in relative to mitochondria.
Chloroplast seems to be immune to HGT, and the only reported case of chloroplast HGT may actually represent transfer between mitochondria of two different plants.
Plastid genes' HGT in nuclear genome are found in one case, in which one of the two copies of plastid gene rps2 in Phelipanche comes from Orobanche type. Putative transfer of atpA/atp1 between chloroplast and mitochondria and mitochondria gene of plastid origin demonstrate that once transferred to mitochondria, these genes can spread in the genome via efficient mitochondria HGT. While most of the known transfers
463

have created a state of gene 'duplication', some chimeric
HGT occurred, generating a hybrid gene that incorporated native and foreign genes. Previous work has reported mtDNA pseudogenes that are transcribed and edited, transcribed but not edited, or not transcribed at all, indicating that most transfers preserved and discovered are neutral, but some genes are transcribed and edited and are potentially functional. HGT may facilitate developmental coordination between parasites and their hosts and it is associated with the formation of host specificity. Recent work indicates that the acquisition of coxI intron could represent a marker of a genomically more widespread historical transformation.
This review will examine evidence for HGT in plant mitochondria, chloroplast and nuclear genomes and the potential mechanisms and consequences of plant-to-plant mitochondrial HGT.
P0369 – ePoster
1
Molecular evolution of water channel proteins gene family in rice
Chiang, Y-C 1 , Wu, C-T 2
Dept of Biological Sciences, National Sun Yat-Sen
University, Taiwan; 2 Institute of Biotechnology, National
Pingtung University of Science and Technology, Taiwan
From 19th to 21st centuries, humans have depended on fossil fuels to create the world. However, the feedbacks including global warning, environmental change and drought problem make the water scarcity in different areas of the world. To develop high drought resistant crops is a method to solve the problem of water scarcity.
Rice is an important major crop in the world and feeding more than half human population. Cultivated rice, Oryza sativa , belongs to family Gramineae. According to grow habitats, cultivated rice separates to two major systems including upland and lowland rice. In world agriculture, humans choose lowland rice system as the major agricultural type. In this system, it needs flooding in the growing season and wastes a lot of water to grow the lowland rice. However, the upland rice keeps more resistant phenotypes to survive in drought environments and grows from plain to 2500 meter high mountain areas.
Meanwhile, the upland rice is a specific system in the cultivated rice and could survive in low support environment areas. It could defense the drought, but the mechanism of drought resistance is still unclear. Water channel protein, same as aquaporins, is a kind of membrane protein to control water pumping between membranes. In this study, we clone and sequence different gene copies of water channel protein from upland and lowland cultivated rice, wild rice, and related species of Oryza genus. By genomic scanning, 11 copies of water channel proteins were found from rice whole genome data. We examine the sequence variations and the selective forces in different gene copies showed different patterns between copies. For example, the locus of 9th chromosome was low nucleotide diversity and divergence within lowland, upland cultivated and wild rice. There are 2, 2, and 7 haplotypes detected from upland, lowland, and wild rice, respectively. It could be affected by purifying selection on the water channel protein locus of 9th chromosome.
P0373 – ePoster
Molecular basis of crassulacean acid metabolism
(CAM) evolution inferred from gene family structures in Neotropical orchids (subtribe Oncidiinae)
Gulle, B 1 , Albion, BL 1 , Silvera, K 2 , Cushman, JC 1
1 Dept of Biochemistry and Molecular Biology, University of Nevada, USA; 2 Dept of Botany & Plant Sciences,
University of California, USA
Crassulacean acid metabolism (CAM) is an important water-conserving photosynthetic pathway present in approximately 7% of vascular plant species from at least
35 families. The Orchidaceae is arguably the largest family of vascular plants with about 30,000 species of which approximately 40% are likely to exhibit some degree of weak or strong CAM. Phylogenetic analysis from 1,103 species native to Panama and Costa Rica revealed that CAM has evolved independently several times within the Orchidaceae. The Oncidiinae, which is the second largest orchid subtribe, was selected for evolutionary analysis, because it has one of the most well established molecular phylogenies available and has been sampled by leaf tissue carbon isotope composition. To better understand the CAM evolution, molecular changes were observed using microarray analysis. Roche/454 pyrosequencing for the strong CAM orchid species
Rossioglossum ampliatum resulted in a total of 189 Mb of data with more than 41,000 contigs and 100,000 singletons. More than 39,000 probe sets were used to design Nimblegen oligonucleotide microarray to detect the expression differences among three closely related orchid species performing photosynthesis in a range of
C to weak CAM to strong CAM. In order to test the
3 hypothesis that CAM evolution is accompanied by gene duplication events and recruitment of discrete isogenes with enhanced mRNA expression, we have investigated gene family structures for phosphoenolpyruvate carboxylase (PEPC), carbonic anhydrase (CAH) and glucose-6-phosphate/Pi translocator (GPT). We have sampled over 2000 cDNA products using a degenerate
RT-PCR strategy to recover an average of five PEPC, four CAH and two GPT isogenes within each family across ten orchid species. A 24-hour gas exchange analysis showed that weak CAM species exhibit limited amounts of nocturnal CO
2
uptake when compared to strong CAM species. Our current results indicate that the evolutionary progression from the ancestral C
3 state to the derived CAM state is accompanied by selective recruitment of discrete isogenes that show increased mRNA expression in order to fulfill the enhanced metabolic demands of CAM to perform nocturnal CO
2 fixation.
P0374 – ePoster
Functional genomics of CAM in the monocot biofuel feedstock crop
Agave sisalana
Boxall, SF 1
Hartwell, J 1
, Waller, JL 1 , Gregory, R 1 , Hall, N 1 ,
1 Dept of Plant Sciences, Institute of Integrative Biology,
University of Liverpool, UK
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Climate change predictions based on a variety of models suggest that the land area covered by arid and semi-arid regions will expand this century as the climate warms and extreme weather events become more frequent.
Furthermore, there is a growing realisation that the human population will grow to 9 billion by 2050 and that the yield of the world's major food crops has reached a plateau. There are thus grave concerns about the ability of the world's farmers to generate sufficient food to sustain the predicted population. At the same time, concerns that global warming is being driven by CO
2 released by the burning of fossil fuels are driving a push to develop sustainable, carbon-neutral liquid biofuels from crop biomass. Current biofuels largely rely on food crop species such as sugar cane and maize, and thus compete with food security for available land. With all of these major mounting pressures, there is a growing acceptance that 21st century agriculture needs to think outside the box in order to respond to the challenges and balance food security with the production of crop feedstocks for biofuels. Crassulacean acid metabolism
(CAM) is a metabolic adaptation of photosynthesis that increases water use efficiency relative to C
3 photosynthesis by between 6 and 20-fold. Thus, CAM plants are able to thrive on arid and semi-arid land where major crops such as rice and wheat will not grow productively. Certain CAM crops can achieve very respectable yields of up to ~ 45 tonnes per hectare per year on seasonally-dry land. In particular, high productivity species of the monocot CAM genus Agave, such as Agave tequilana, Agave fourcroydes, and Agave sisalana, have been proposed as excellent biofuels feedstocks crops suited to seasonally-dry lands. A key reason why Agaves are believed to have great potential is their high content of fructan sugars which can very readily be fermented into ethanol. In addition, they have a high percentage of low lignin cellulose (e.g. sisal fibre), which could be exploited with second-generation biofuels technology aimed at the conversion of cellulose to ethanol. Despite the urgent and pressing need to optimize the exploitation of certain CAM plants for human good, they remain one of the most poorly understood photosynthetic types and little scientific research or breeding has been done to enhance the productivity of CAM crops. Our goal is to develop and exploit genomic resources for CAM species. In particular, we are using second-generation high throughput DNA sequencing to decode the transcriptomes and genomes of CAM species as a basis for discovering all of the genes required for efficient
CAM. We focus on genes involved in the optimisation of the biochemical steps of CAM over the light dark cycle, particularly on components of the circadian clock signalling pathway linking CAM biochemistry to the central molecular circadian clock. This poster will present an overview of our recent progress in understanding the functional genomics of CAM in our chosen biofuels feedstock crop, Agave sisalana .
P0375 – ePoster
Using RNA-seq to dissect the molecular basis for circadian optimisation of crassulacean acid metabolism
Boxall, SF 1 , Dever, LV
N 1 , Hartwell, J 1
1 , Gregory, R 1 , Knerova, J 1 , Hall,
1 Dept of Plant Sciences, Institute of Integrative Biology,
University of Liverpool, UK
The goal of our research is to identify all the genes involved in the establishment and circadian coordination of crassulacean acid metabolism (CAM) in the obligate
CAM species, Kalanchoë fedtschenkoi. K. fedtschenkoi is an excellent model for the study of CAM photosynthesis for a number of key reasons. Firstly, extensive biochemical and physiological characterization of CAM has previously been performed with this species.
Secondly, we have developed a simple, rapid and efficient stable transformation system. Thirdly, it is readily amenable to molecular, biochemical and physiological analysis permitting detailed phenotypic characterization of transgenic lines. Finally, it performs a clear developmental progression from C to CAM during
3 leaf ontogeny. By performing RNA-seq experiments using both the Roche 454 and Applied
BiosystemsSOLiD second generation sequencing systems, we have compared the transcriptome of C and
3
CAM leaves from K. fedtschenkoi over a diurnal cycle.
Detailed bioinformatic analysis of the RNA-seq data, has permitted us to identify thousands of known and unknown genes with potential roles in either the biochemical steps or the circadian coordination and optimization of CAM. We are focusing downstream analysis on genes encoding regulatory proteins that could play a role in the circadian coordination of CAM; undertaking detailed RT-PCR studies to corroborate the
RNA-seq data, and then manipulating putative circadianregulated CAM genes in transgenic lines using both silencing with RNAi, and constitutive over-expression.
In addition, we have also silenced and over-expressed many of the known major metabolic steps of the CAM pathway (PEPc, PPDK, NAD(P)-ME etc), and have begun detailed phenotypic analysis of these transgenic lines. Some of our preliminary phenotypic analysis of the transgenic lines will be presented, along with an overview of the discoveries achieved through the RNAseq analysis of the diurnal and circadian control of the
CAM transcriptome.
P0377 – ePoster
1
The
Amborella
Genome Sequencing Project: an evolutionary reference sequence
Der, JP 1
S 2
, Ralph, P
, Walts, B 2
1 , Ma, H 1 , DePamphilis, C
, Barbazuk, WB 2 , Soltis, D 2
1 , Chamala,
, Soltis, P 2
Dept of Biology, Pennsylvania State University,
University Park, USA; 2 Dept of Biology, University of
Florida, Gainesville, USA
The origin and early diversification of flowering plants
(angiosperms) had profound impacts on Earth's biota, providing the raw genetic material from which most crops and economically important plants were derived.
The diversification of genes, genomes, and important traits cannot be adequately interpreted without a comparative framework firmly rooted with genome sequences from basal angiosperms. As the sister species to all other extant flowering plants, Amborella trichopoda holds a singular position in the flowering tree of life for establishing this comparative genomics framework. The Amborella Genome Sequencing Project
465

is an NSF funded collaborative project that seeks to produce a high quality finished genome sequence, complete with an accurate chromosome-scale physical map and evidence based gene and transposable element annotations. We are combining the latest multi-platform sequencing technologies with traditional and cuttingedge physical mapping techniques to produce a reference sequence ideally suited for comparative evolutionary analyses to shed light on genomic characteristics of the the last common ancestor of extant angiosperms and the evolution of gene content and genome structure throughout angiosperm history. To leverage this genome sequence, we are also developing new bioinformatic tools and a public access website to make this important resource available to the scientific community.
P0378 – ePoster
Genetic basis of plant architectural development in grasses – insights from Foxtail Millet
Doust, A 1 , Mauro-Herrera, M 1 , Malahy, M 1 , Stromski, J 1
1 Oklahoma State University, Botany Dept, Stillwater,
OK, USA
The breeding of efficient plant architectures for biofuel production requires the understanding of the genetic basis of plant architecture. In grasses, plant architecture can be decomposed into branching structure modules and accompanying morphological and anatomical differentiation. Comparative genomics has allowed information from maize and other model systems to be applied to other grass systems, such as the foxtail millet - green millet ( Setaria italica – S. viridis ) system. We are particularly interested in this system because of its pronounced variation in branching patterns, and because of completed genome sequences for both species.
Multiple trials of mapping populations from a cross between green and foxtail millet have indicated major
QTL for branching and height, some co-occurring with candidate genes such as the teosinte branched1 and barren stalk1 orthologs. A recent set of greenhouse and field trials with an F7 RIL population recapitulates those results, but finds that QTL can vary between different growth stages, suggesting that some genomic regions have a constant influence in one or more traits during the life span of the plant while others seem to be time/developmental stage specific. Our results provide insights into the tradeoffs taking place throughout the development of plant architecture during the plant's life span, suggesting that mechanisms of genetic control of branching phenotypes vary over the life cycle of the plant.
P0380 – ePoster
Wounding, drought and cold stresses regulate wheat
Wcor410b gene expression through the same Crepeat element
Eini, O 1 , Yang, N 1 , Pyvovarenko, T 1 , Bazanova, N 1
, Langridge, P 1 , Hrmova, M 1
,
, Tikhomirov, N 1 , Eliby, S 1
Lopato, S 1
1 ACPFG, University of Adelaide, Australia
466
Dehydrins are one of the major groups of late embryogenesis-abundant (LEA) proteins. The promoters of these genes are highly activated in flowers and vegetative tissues under stress conditions. This makes the promoters of dehydrins potentially good candidates for specific stress-inducible expression in transgenic plants of genes that may increase tolerance to abiotic stresses.
However, many of the LEA promoters activated by a broad spectrum of stresses and stimuli have high basal levels of activity. This reduces their value for genetic engineering. To assess the possibility of ‘improvement’ of LEA promoters, we characterised their structure and function. TdCor410b promoter was isolated and used for mapping of the functional CRT element in transient expression assays in wheat cell cultures. Tandem repeats of the core of the functional CRT (GCCGAC) element, with and without adjacent sequences, were used as baits in yeast one-hybrid (Y1H) screens for the isolation of
TFs from cDNA libraries prepared from developing grain or whole spikes of unstressed or drought/cold stressed wheat and barley plants. Screens resulted in the isolation of Drought Responsive Element Binding (DREB) factors and three different subfamilies of Ethylene Responsive
Factors (ERFs). Activation of the TdCor410b promoter by some of the isolated transcription factors has been confirmed using transient expression assays in wheat cell cultures. Analysis of spatial and stress inducible expression of the isolated ERFs suggests that TaERF4a is the best candidate for the regulation of Wcor410b during plant development and under drought stress. Some of the isolated ERF genes could also be involved in regulation of the Wcor410b gene under cold stress and wounding.
Specificity of binding of isolated TFs to different ciselements was demonstrated. A series of threedimensional models of ERFs in complexes with DNA oligomers (cis-elements) was generated. These models described the specificity of interactions of the AP2 domains with the GCCGAC (CRT), ACCGAC (DRE) and GCCGCC (GCC-box) core elements. It was demonstrated that TaERF4a-like ERFs bind only CRT elements but were unable to bind the GCC-box. The amino acid residues responsible for the specificity of binding were identified and confirmed experimentally by substitution of Pro42 for Arg, which restored the
TaERF4a interaction with the GCC-box. The presence of a single promiscuous CRT cis-element in the shortest version of the TdCor410b promoter (275 bp) permitted generation of several artificial promoters by the exchange of CRT for several other stress-related cis-acting elements. These substitutions could potentially decrease basal levels of promoter activity and/or increase specificity of the promoter. An analysis of the activity and specificity of artificial promoters is in progress.
P0381 – ePoster
Variability and genetic differentiation by ISSR markers in
Mimosa dolens
(Leguminosae)
1
Bessega, C 1,2 , Morales, M 2 , Fortunato, R 3
CONICET, Universidad de Buenos Aires, Argentina;
Argentina; 3 INTA, Argentina
2
Mimosa dolens (Leguminosae) constitutes a morphological complex growing in tropical and subtropical regions of southern South America. Within

Subserie Dolentes intra-specific taxa were proposed in
M. dolens . Previous morphological and cytological studies indicated that M. dolens subsp. rigida and acerba ,
(NE from Argentina, Paraguay and S of Brasil); M. dolens subsp. acerba var. rudis (4x); M. dolens subsp. rigida and M. dolens subsp. callosa (8x) (from NE of
Paraguay) lives in sympatry and intermediate individuals between them can be found. The objective of this study was to analyze the genetic variability, population structure and phenetic relationships among Mimosa dolens populations using ISSR (Inter Simple Sequence
Repeat) markers in order to help in the understanding of these complex taxa. DNA of 34 seedling from two different populations from Paraguay were analyzed. A total of 81 loci were scored using three ISSR primers and analyzed with AFLP-SURV software. A high level of genetic variability was found in San Pedro and Porteira
Ortiz populations (He = 0.19–0.29; P = 33% –86% respectively). The analysis of genetic structure indicated that the between population genetic diversity (37.6 %) was lower than the within population genetic diversity
(62%). Genetic differentiation among populations (FST =
0.3687) was highly significant. The UPGMA analysis based on DICE coefficient (NTSYSpc 2.0) grouped the individuals from San Pedro population but split the individuals from Pereira Ortiz. The PCA analysis
(performed with GenAlEx 6) explained the 86% of the variation in the 3 first axis and is consistent with phenogram obtained. The relationships can be explained by the morphological criteria and in those cases where external traits makes difficult the classification the ISSR technique can help in the delimitation of the taxa. Future experiments including more populations and loci can be performed to find diagnostic loci.
P0382 – ePoster
The effects of light and temperature on cyanogenesis in forage sorghum
Fromhold, S 1
R 1 , Hamill, J 1
, O'Donnell, N 1
, Neale, A 1
, Blomstedt, C 1 , Gleadow,
1 Monash University, Australia
Sorghum is a C
4
, drought-resistant plant species, which is also cyanogenic in all tissues except the mature seeds.
The cyanogenesis pathway in sorghum produces the stable cyanogenic glucoside, dhurrin. When the plant tissue is disrupted the dhurrin is hydrolysed to release hydrogen cyanide (HCN). At low concentrations HCN can be metabolised safely by mammals but at higher concentrations it is toxic as it causes metabolic asphyxiation. The HCN potential (HCNp) of sorghum is important in the cattle industry as forage sorghum is used as an alternate feed crop over summer. Environmental factors, such as drought, are known to increase the HCNp of sorghum but little is known regarding the affects of other conditions, including light and temperature fluctuations. Analysis of the sequenced promoter regions as well as putative promoter regions of the three biosynthetic genes in the cyanogenesis pathway;
CYP79A1, CYP71E1 and UGT85B1, showed that in all three sequences there are potential light, circadian and temperature response elements. It has been postulated that sorghum may also go through a circadian or diurnal rhythm in relation to its cyanogenesis pathway. Multiple experiments have been conducted to compare different light treatments on hydroponically grown forage sorghum that were and placed in cabinets with controlled temperature and nutrients. The plants were subjected to constant temperature or thermocycling temperature and three light treatments; continuous light, continuous darkness or a 14/10 light/dark cycle, with plants harvested every four hours for 60 hours. These experiments were designed to determine if there is a circadian or diurnal effect on the cyanogenesis pathway in forage sorghum and the results will be discussed.
P0383 – ePoster
Small-scale genetic structure among patches and populations of
Mercurialis annua
L. with contrasting
1 sexual systems, as revealed by amplified fragment length polymorphism (AFLP)
García-Castaño, JL 1,2 , Korbecka, G 3 , Pannell, J 1
Dept of Plant Sciences (University of Oxford), Oxford,
UK; 2 Departamento de Biología Vegetal y Ecología
(Universidad de Sevilla), Sevilla, Spain; and Plant Cultivation), Puawy, Poland
3 Dept of Plant
Breeding and Biotechnology (Institute of Soil Science
The advent of rapidly mutating molecular markers such as microsatellites has allowed the detailed description of within-population gene flow for a large number of species. These studies have typically revealed that most mating occurs among spatial neighbours, but also that mating frequently occurs over substantial distances, i.e., mating in any one generation is often far from panmictic.
Most studies of within-population gene flow have been conducted on long-lived woody perennials that are either hermaphroditic or dioecious. Here, we present novel analyses of the within-population spatial genetic structure of an androdioecious annual plant, in which males coexist with hermaphrodites. We sampled plants from within six populations from an androdioecious region in southern Spain that ranged in their frequency of males and used amplified fragment length polymorphism
(AFLP) to compare measures of genetic diversity and differentiation at small spatial scales of several meters.
Our results point to the importance of within-population demographic processes in shaping patterns of genetic variation.
P0385 – ePoster
Functional and comparative genomics of CAM in the dicot model Kalanchoëfedtschenkoi and the monocot biofuel feedstock crop
Agave sisalana
Boxall, SF 1 , Dever, LV
N 1 , Hartwell, J 1
1 , Gregory, R 1 , Knerova, J 1 , Hall,
1 University of Liverpool, UK
Climate change predictions based on a variety of models suggest that the land area covered by arid and semi-arid regions will expand this century as the climate warms and extreme weather events become more frequent.
Furthermore, there is a growing realisation that the human population will grow to 9 billion by 2050 and that the yield of the world's major food crops has reached a
467

plateau. There are thus grave concerns about the ability of the World's farmers to generate sufficient food to sustain the predicted population. At the same time, concerns that global warming is being driven by CO
2 released by the burning of fossil fuels are driving a push to develop sustainable, carbon-neutral liquid biofuels from crop biomass. Current biofuels largely rely on food crop species such as sugar cane and maize, and thus compete with food security for available land. With all of these major mounting pressures, there is a growing acceptance that 21st century agriculture needs to think outside the box in order to respond to the challenges and balance food security with the production of crop feedstocks for biofuels. Crassulacean acid metabolism
(CAM) is a metabolic adaptation of photosynthesis that increases water use efficiency relative to C
3 photosynthesis by between 6 and 20-fold. Thus, CAM plants are able to thrive on arid and semi-arid land where major crops such as rice and wheat will not grow productively. Certain CAM crops can achieve very respectable yields of up to ~ 45 tonnes per hectare per year on seasonally-dry land. In particular, high productivity species of the monocot CAM genus Agave , such as Agave tequilana, Agave fourcroydes , and Agave sisalana , have been proposed as excellent biofuels feedstocks crops suited to seasonally-dry lands. A key reason why Agaves are believed to have great potential is their high content of the fructan sugars which can very readily be fermented into ethanol. In addition, they have a high percentage of low lignin cellulose (e.g. sisal fibre), which could be exploited with second-generation biofuels technology aimed at the conversion of cellulose to ethanol. Despite the urgent and pressing need to optimize the exploitation of certain CAM plants for human good, they remain one of the most poorly understood photosynthetic types and little scientific research or breeding has been done to enhance the productivity of CAM crops. Our goal is to develop and exploit genomic resources for CAM species. In particular, we are using second-generation high throughput DNA sequencing to decode the transcriptomes and genomes of CAM species as a basis for discovering all of the genes required for efficient
CAM. We focus on genes involved in the optimisation of the biochemical steps of CAM over the light dark cycle, particularly on components of the circadian clock signalling pathway linking CAM biochemistry to the central molecular circadian clock. This talk will present an overview of our recent progress in understanding the functional genomics of CAM both in our model dicot system, Kalanchoë fedtschenkoi , and our chosen biofuels feedstock crop, Agave sisalana .
P0386 – ePoster
Functional genomics of a generalist parasitic plant
Honaas, L 1
Altman, NS
, Wickett, N
Taylor, CG
2
5
, Bao, Y
, Yoder, JI 4
DePamphilis, CW 1
1 , Wafula, E 1 , Zhang, Y 1
3 , McCombie, WR 7 , Schuster, S
, Landherr, L 1
, Der, J 1
6
,Wu, B 4
1 Dept of Biology and Huck Institutes For The Life
Sciences, Pennsylvania State University, University Park,
USA;
USA;
2
3
Dept of Statistics, Pennsylvania State University,
Dept of Microbiology, DNA Sciences Core,
University of Virginia, Charlottesville, USA; 4 Dept of
,
,
,
468
5
Vegetable Crops, University of California, Davis, USA;
Dept of Plant Pathology, Ohio State University,
Wooster, USA; 6 Dept of Biochemistry and Molecular
Biology, Pennsylvania State University, USA; 7 Cold
Spring Harbor Laboratory, Watson School of Biological
Sciences, Cold Spring Harbor, New York, USA
The Orobanchaceae are a plant family whose parasitic members growing in Africa and the Mediterranean include the devastating agricultural pests, witchweed
( Striga ) and broomrape ( Orobanche ). In addition to the pressing socioeconomic issues surrounding these parasites, this family represents a unique opportunity to study parasite biology since its members span varying degrees of parasitism from feel-living to nonphotosynthetic, obligate parasites. The Parasitic Plant
Genome Project (PPGP, http://ppgp.huck.psu.edu/ ) aims to discover the genetic changes in the Orobanchaceae that allowed the transition from autotrophy to heterotrophy by transcriptome analysis of various tissue and life stage-specific samples. Analysis via Next
Generation Sequencing (NGS) technology is allowing the discovery of genes being used by the parasite in each specific sample and builds a comparative framework that will allow identification of genes involved in the parasitic lifestyle. Despite the great power of this approach, gene expression patterns of highly specialized cells can become diluted since they may represent a small percentage of the total tissue harvest. We are using laser capture micro-dissection to sample tissues in a highly cell-specific manner, allowing us to capture cells of interest, extract and then amplify exceedingly small RNA samples, sequence them with NGS technology and analyze them without the aid of prior sequence knowledge. Using this method we analyzed the hostparasite interface transcriptome of a generalist member of the Orobanchaceae, Triphysaria versicolor .
P0387 – ePoster
Use of D1/D2 region in 28S rDNA as the DNA barcode for wood rot fungi
Horisawa, S 1 , Maeda, M 1
1 Kochi University of Technology, Japan
The identification of wood-rot fungi is not easy because they lives in the hidden place such as in wood, their mycelia lack recognizable distinction, and morphologic character of their fruit body has a wide variation. DNA barcoding is a taxonomic method that uses a short genetic marker in an organism's DNA to identify it as belonging to a particular species. The internal transcribed
(ITS) spacer region of nuclear rDNA has been used as a target for analyzing fungal diversity in environmental samples, and has recently been selected as the standard marker for fungal DNA barcoding. However, the genetic information for DNA barcoding of wood rot fungi has not been sufficient. The use of the ITS region for the identification and discrimination of wood rot fungi was investigated in the previous study. The result showed that the ITS was generally useful for DNA barcoding but more reliable marker should be explored. The D1/D2 region in 28S rDNA was employed in the present study.
The 50 strains of wood-rot fungi were selected from the fungal collection of NITE Biological Resource Center

(NBRC), Japan. Genomic DNA was extracted from fungal colony on a plate medium. The D1/D2 region was amplified by PCR using primer NL1 and NL4 and subcloned into the plasmid vector for sequencing. The sequence of each strain was compared with information in GenBank/EMBL/DDBJ database by BLAST search and was checked species of the strain. More information about species was obtained from cluster analysis. The length of D1/D2 region fragments of the test fungi was
618-647 bp, that was similar the full length of the ITS.
The D1/D2 sequences of most species of the test strains were referable in the biological database of
GenBank/DDBJ/EMBL and identifiable the species.
Results showed that development of identifying tool may be possible for wood degradation study. A series of sequence from ITS to D1/D2 would be more informative to identify the species.
P0388 – ePoster
Plant comparative genomics tools from Gramene database
Jaiswal, Pankaj 1
1 Oregon State University, USA
Newly completed plant genome sequences has empowered researchers from plant biology, breeding, phylogenomics, genetics and computational modeling interests to extensively use these genomes as reference maps to guide their research on modeling genome-wide associations to find candidate genes and their associations to phenotypes of agronomic importance.
Similarly the current generation of systems biology projects is looking at exploiting these datasets for building gene-to-gene networks and their association to environmental conditions. Before developing the analysis tools to do these types of studies, the data must be fetched from in-house and/or from remote data archives and aggregated into databases. In this direction, we have created the Gramene database (http://www.gramene.org).
It is a comprehensive resource for comparative functional genomics in plants which allows integration of rich annotations such as, phenotypes and genetic variation with maps that link physical and genetic positions within and across species. The database hosts annotated genomes of Oryza species, Zea mays, Sorghum bicolor,
Brachypodium distachyon, Arabidopsis thaliana and lyrata , Vitis vinifera , and Populus trichocarpa , as well as genetic maps from over 30 intra- and/or inter-specific crosses. It also holds population diversity data from large-scale SNP genotyping studies, including those of rice, maize and Arabidopsis , some of which include phenotypic data. Functional information is provided by
Quantitative Trait Loci (QTL), protein domain annotation, Gene, Trait and Plant Ontologies, and biochemical pathway databases. Evolutionary context and inference of orthologs and paralogs is provided by phylogenetic analysis built from multiple alignments of protein sequences from fifteen species, including five non-plant models. Sequenced whole genome alignments and synteny maps promote inter-species browsing and simultaneous viewing of multiple species over conserved regions. Seamless integration of these data with searching and visualization tools provides a powerful discovery platform using both forward and reverse genetics approaches. In this presentation, we will provide an overview of the (1) latest developments in the field of plant genome informatics, (2) the Gramene database and
(3) use cases and examples to show the informatics platform we have developed.
P0389 – ePoster
Distinguishing genetic individuals in the clonal shrub
1
Tetratheca juncea
Sm. (Tremendraceae)
Jones, A 1 , Grace, E 1 , Castor, C 1 , Driscoll, C 1 , Cole, M 1
Centre for Sustainable Ecosystem Restoration, School of
Environmental and Life Sciences, University of
Newcastle, Australia
Tetratheca juncea is a vulnerable shrub (NSW
Threatened Species Act, 2000) restricted to 200 km of coastal woodland around the Hunter Region. It reproduces sexually and clonally via rhizomatous growth. The clonality of T. juncea makes it impossible to distinguish genets and ramets based on current visual surveying methods, where population sizes are estimated by defining all the stems within a 300mm radius as one
‘clump’, which is in turn counted as one individual plant.
If populations cannot be counted accurately, the impact of population removal and the equivalency of related offsets cannot be properly assessed. As T. juncea occurs on costal land that is subject to competing interests such as development, mining and conservation, its long-term survival is contingent on prioritising, and understanding the impacts of, population removal. It is therefore necessary to develop a surveying method that reflects the true number of genetic individuals in a population.
Genetic information is also needed to effectively manage reserved populations – for example, in understanding the evolutionary status and ecological requirements of the plant’s mixed mating system. This project has resulted in the development of a sensitive, high-throughput molecular technique for generating DNA profiles
(genotypes) that can distinguish even highly related genets. These genotypes are compared by analysing differences in the lengths of polymerase chain reaction
(PCR)-amplified fragments from microsatellite markers.
PCR of these microsatellites, which have large amounts of DNA sequence variation, generates fragments of different sizes according to the number of nucleotide repeats in the DNA sequence of a particular individual.
Ten primer sets, which were developed for another species of Tetratheca by McPherson et al. (2008), have been successfully tested in T. juncea . These ten markers have been multiplexed for PCR and fluorescently labelled using the M13 method described by Schuelke
(2000) for efficient, cost-effective fragment analysis.
Several samples have been genotyped, including three from the same 300mm area ‘clump’. In both the clumps sampled this way, two individuals were distinguished from the three samples taken. This indicates that there may be much more fine-scale genetic diversity in T. juncea than previously thought, which is overlooked in current plant surveys. The merit of this technique in population-level genetic analysis will be tested in a study population located in a semi-urbanised area in Lake
Macquarie. This information will be used to relate the spatial and genetic structuring of sampled plants. It will also be used to generate metrics of gene flow and
469

population genetic health such as inbreeding and heterozygosity levels. Future work in T. juncea using the techniques developed in this project includes paternity analysis, tracking pollinator movement and seed dispersal, and between-population genetic structuring.
P0391 – Poster
Variation of DNA contents in
Primula jesoana
1
Kanehara, R 1 , Hayashi, M 1
Nogami, T 4 , Wagatsuma, T 5
, Ikeda, M
, Kato, J 6
2 , Honda, K 3
, Mii, M 1 , ,
,
Graduate School of Horticulture, Chiba University,
Japan; 2 Bioscience and Biotechnology Center, Nagoya
University, Japan; 3 Fuculty of Agriculture & Life
4
Science, Hirosaki University
Hakusan Nature Conservation Center, Japan;
Gakuen University, Japan;
University of Education, Japan
6
5 Rakuno
Dept of Biology, Aichi
Primula jesoana is a native species in Japan and two kinds of chromosome number, 2n=24 and 26, had been reported. Previous our flow cytometric analysis by DAPI staining using cultivated Primula jesoana indicated that relative DNA content varied among each plant.
Moreover, although six kinds of chromosome number,
2n=24, 25, 26, 27, 28 and 29, were found among 81 cultivated Primula jesoana by cytogenetic observation, there were not correlations between DNA content and chromosome number. Because additional small chromosomes like B chromosomes could be observed in karyotypes of 2n=25, 26, 27, 28 and 29, crosses between plants with different karyotypes were performed to trace transmittance of these small chromosomes. Because chromosome numbers among progenies were observed between 2n=24 and 32 and additional small chromosomes did not transmit under Mendelism, these results were concluded that these additional small chromosomes were B chromosome. Flow cytometric analysis using 9 natural population of Primula jesoana in
3 regions of Hokkaido revealed that differences of DNA content were detected from 5.9% to 12% within each population and the difference of DNA content between the minimum plant and the maximum one was detected at 18.6% among all plants analyzed. These results indicated that Primula jesoana was a species with unstable DNA content and B chromosome.
P0392 – Poster
2
1
Variation of DNA content in
Primula tosaensis
Kato, J
A 1
M 3
1 , Tanaka, N 2
, Ito, N 1
Dept of Biology, Aichi University of Education, Japan; and Sciences, Japan;
, Hayashi, M
, Otsuka, K 5
Tsukuba Botanical Garden, National Museum of Nature
3
3 , Ikeda, M 4
, Ohashi, H 6
, Matsuda,
, Serizawa, S 1 , Mii,
Graduate School of Horticulture,
Chiba University, Japan; 4 Bioscience and Biotechnology
Center, Nagoya University, Japan; 5 Nagano
Environmental Conservation Research Institute,
Japan; 6 Faculty of Agriculture, Ehime University, Japan
Primula tosaensis is a native species in Japan, distributed from west Kanto to Kyushu and have a variety, P.
470 tosaensis var. brachycarpa , which distributed into
Southern Japanese Alps. Previous our flow cytometric analysis by DAPI staining using cultivated Primula tosaensis revealed that there was about 25% difference of
DNA content between P. tosaensis and P. tosaensis var. brachycarpa , regardless of the same chromosome number, 2n=2x=24. In this study, flow cytometric analysis was conducted using mainly natural populations of P. tosaensis and P. tosaensis var. brachycarpa . The
Bonferroni test (P‹0.01) showed significant differences among populations. Populations could divide into six groups in order to low DNA content; (1) Primula tosaensis var. brachycarpa ’Kamanashikozakura’, (2)
Primula tosaensis var. brachycarpa 'Shinanokozakura’,
(3) Primula tosaensis from Gifu and Mie, (4) Primula tosaensis from North of Tokushima and Miyazaki, (5)
Primula tosaensis from Kochi and (6) Primula tosaensis from South of Tokushima. The relative DNA content within two populations of P. tosaensis var. brachycarpa varied 1.16-fold, while those of DNA content within eight populations of P. tosaensis varied 1.40-fold.
Maximum difference among all populations was detected as about 60%. In the results of flow cytometric analysis, the relative DNA contents tended to increase from east to west. These results were considered that the DNA content might be varied at the migration time of this species. Phylogenetic analysis is now in progress. The relationships between DNA contents and phylogenetic relationships will be discussed.
P0393 – ePoster
A robust method maximising data output for transcriptome analysis using GS FLX 454
1
Kay, PT 1 , Lawless, K
Spangenberg, G 1
1 , Tibbits, J 1 , Hayden, M 1 ,
Departmant of Primary Industries Victoria, Bundoora,
Australia
Next generation sequencing instruments have greatly increased sequence output enabling direct molecular genetic analyses by sequencing e.g. whole transcriptome analysis. Such experiments were largely not possible due to cost in the past. The Roche GS FLX 454 instrument typically yields around 400 Mb of sequence data from a single run. While this is small in comparison to Illumina genome analysers, the GS FLX 454 can generate longer reads (300–350bp), which is advantageous for de novo sequence assembly, especially in polyploid plant species.
We have developed a robust protocol that maximises GS
FLX sequence output and enables the sequencing of full length transcripts for transcriptome analysis. The method is robust having been tested across multiple libraries from various indigenous and exotic plant species. This protocol could also be easily adapted for sequencing other nucleic acids such as genomic DNA and amplicons.
Our method builds on the Meyer et al (2009) BMC
Genomics method and consists of cDNA synthesis using a modified Epicentre ExactSTART„· Full-Length cDNA
Library Cloning Kit protocol, cDNA fragmentation and ligation of GS FLX compatible adaptors, size selection and PCR amplification of internal or external (3- and 5- termini) cDNA fragments with a specific adaptor configuration, which are used directly for emulsion PCR.
There are three major benefits of this method: 1)

sequence coverage along the entire length of the transcripts is more even, 2) library preparation is simplified, as amplified double stranded cDNA can be used directly for emulsion PCR, and 3) high sequence output (500–550Mb from a single run) and longer reads
(median length of 400–450bp) are achieved.
P0394 – ePoster
Parentage determination of
Vanda
Miss Joaquim
(Orchidaceae) through two chloroplast genes rbcL and matK
Khew, GS 1 , Chia, TF 2
1 Singapore Botanic Gardens, Singapore; 2 Natural
Sciences and Science Education AG, National Institute of
Education, Nanyang Technological University,
Singapore
Background and Aims – The popular hybrid orchid
Vanda Miss Joaquim was made Singapore's National flower in 1981. It was originally registered with the
Royal Horticultural Society in 1893, as a cross between
Vanda hookeriana and Vanda teres . However, no record had been kept as to which parent contributed the pollen.
This study was conducted using DNA barcoding techniques for the resolution of hybrid parentage and to determine the pod parent of V.
Miss Joaquim, thereby inferring the pollen parent of the hybrid.
Methods – Two chloroplast genes, mat K and rbc L, from five related taxa, V. hookeriana, V. teres var. alba , V. teres var. andersonii , V. teres var. aurorea and V.
Miss
Joaquim 'Agnes' were sequenced. The mat K gene from herbarium specimens of V. teres and V . Miss Joaquim, both collected in 1893 was also sequenced.
Key Results – No sequence variation was found in the
600bp region of rbc L sequenced. Sequence variation was found in the mat K gene of V. hookeriana, V. teres var. alba , V. teres var. aurorea and V. Miss Joaquim 'Agnes'.
Complete sequence identity was established between V. teres var. andersonii and V.
Miss Joaquim 'Agnes'. The mat K sequences obtained from the herbarium specimens of V. teres and V.
Miss Joaquim were completely identical to the sequences obtained from the fresh samples of V. teres var. andersonii and V.
Miss Joaquim
'Agnes'.
Conclusion – The pod parent of Vanda Miss Joaquim
'Agnes' is V. teres var. andersonii and the pollen parent is
V. hookeriana . Despite numerous reproductions of the cross, we have shown that the Vanda Miss Joaquim
'Agnes' available today is the same as that registered in
1893. The mat K gene was more informative than rbc L and facilitated differentiation of varieties of the same species.
P0395 – ePoster
LRT1, a novel retroelement, in the genus
Lilium
, Son, J-H 1 , Lee, S-I 1 , Yie, S-W Park, K-C 1
Kim, N-S 1
1 , Kim, J-H 2 ,
1 Dept of Molecular Biosciences, Kangwon National
2 University, Republic of Korea; Dept of Horticulture,
Kangwon National University, Republic of Korea
Retrotransposons are highly abundant in plant kingdom.
Variable content of LTR (Long Terminal Repeat) retrotransposons causes variation in the C-values in plant species. Species in the genus Lilium have the largest genome in flowering plants. A novel Ty1/copia retrotransposable element, LRT-1 (Lily Retro
Transposon-1), was isolated from L. longifolium . LRT-1 was structured with 243 bp of LTR and Pol gene, but absent Gag gene. The characteristic PBS (primer binding site) and PPT (polypurine track) were flanked at the end of LRT sequences. LRT-1 showed high level sequence similarity with Tnt1 of Tobacco. The LTR sequences of the LRT-1 were highly conserved among the species in the Lilium genus. In Southern blotting analysis using the
LRT sequence as a probe, the LRT-1 was revealed to be present only in Lilium species in the Liliaceae family.
The hybridization signals varied among various Lilium species, indicating that the LRT-1 might have contributed to the difference in genome size in the Lilium species.
FISH analysis showed that the LRT-1 was distributed along the chromosomes rather than localized.
Transposon-display (TD) analysis showed high variation not only within species but also between species in the genus.
P0397 – ePoster
Capricious distribution of a cysteinyl tRNA gene of unknown origin in the mitochondrial genomes of angiosperms: an example of horizontal transfer?
Kubo, T 1 , Kitazaki, K 1 , Kagami, H 1
1
T 1
Hokkaido University, Japan
, Fujita, A 1 , Mikami,
Mitochondrial genomes of angiosperms are the largest one in eukaryotes. Nucleotide sequence of mitochondrial genomes from ~19 plant species have revealed that expansion of the intergenic region is the principal mechanism of the size increasing but repertoire of mitochondrial genes has been rather decreased during the course of their evolution. To date, how the intergenic region has expanded and, if any, the source of them are unclear. On the other hand, emerging evidence indicate that horizontal transfer has been occurred during the mitochondrial genome evolution of angiosperms. In some cases, horizontal transfer was detected as traces of gene conversion-like activity between host- and transferred mitochondrial genes. In another cases, sequence homology to non-plant organisms has suggested their horizontal-transfer origin. Horizontal transfer might have been involved in the expansion of intergenic region. We had determined the entire nucleotide sequence of sugar beet mitochondrial genome and reported that a cysteinyl tRNA gene of unknown origin (tRNA-Cys2) was expressed (Kubo et al. 2000).
We investigated the presence/absence of the tRNA-Cys2 gene in some angiosperms. Database analysis: BLAST search using the tRNA-Cys2 gene as a query detected homologous sequence from cucumber, watermelon, soybean, ground nut and Aquilegia coerulea , but none from the other plants. Detection of tRNA-Cys2 gene:
PCR analysis of 26 angiosperms revealed additional plants having the tRNA-Cys2 gene, Daphniphyllum macropodum, Hibbertia pedunculata, Basella rubra . In addition, molecular cloning of the tRNA-Cys2 gene from
471

spinach was successful. Our quantitative PCR assay revealed that the copy number of the tRNA-Cys2 gene was comparable to cob gene in the above plants. Is tRNA-Cys2 a functional gene? We isolated total RNA from some plants having the tRNA-Cys2 gene under acidic or basic conditions. RNA gel blot analysis of these samples indicated the accumulation of tRNA-Cys2 molecules that were charged with amino acids in sugar beet, spinach and cucumber, whereas no signal was detected from watermelon or soybean. How is the original cysteinyl tRNA gene? In spinach mitochondria, the original cysteinyl tRNA gene (i. e. those descended from the initial mitochondrion. Also known as 'Native'class tRNA) was also present and its transcripts were aminoacylated. However, none of the original tRNA-Cys gene was found in cucumber. In sugar beet, it was a pseudo gene. Possible evolutionary scenarios: because the tRNA-Cys2 sequence was not detected in plant mitochondrial genome other than some angiosperms, horizontal transfer of tRNA-Cys2 gene is very likely. Its disjunctive distribution can be explained by either that the acquisition of tRNA-Cys2 gene in the common ancestor of angiosperm was followed by independent loss of many plant lineages, or that independent acquisition of tRNA-Cys2 in each plant lineage. The tRNA-Cys2 gene can co-exist with the original tRNA-
Cys gene, while either has occasionally lost in some plant lineages.
P0398 – ePoster
Biodiversity Heritage Library for Europe
Kvacek, J 1 , Scholz, H 2
1 National Museum, Prague, Czech Republic; 2 Museum für Naturkunde, Leibniz Institute for Research on
Evolution and Biodiversity at the Humboldt University
Berlin, Germany
An important impediment for the implementation of the
Convention on Biological Diversity (CBD) of the United
Nations is the lack of access to core information on animals and plants. This information is concentrated in the scientific books and journals of the past centuries. At the moment, the only way to access to this knowledge is to visit a number of different libraries. This complicates much of the fundamental research in biological science.
Since 2007, the Biodiversity Heritage Library has been systematically removing this impediment by making this literature available over the Web. To date (January 2010) already 32.5 million pages of almost 87,000 volumes are freely available through http://www.biodiversitylibrary.org. Managed by the
Museum für Naturkunde Berlin, the Biodiversity
Heritage Library for Europe (BHL-Europe) started on
May 1st 2009 within the framework of the EU program eContentplus. It will now further develop, expand, and enhance the Biodiversity Heritage Library by bringing together the extensive collections of biodiversity literature held in major European natural history, botanical, and research libraries. The new interface of the
Portal will be multilingual enabling users to search in their native language. Innovative functionalities facilitates quick access to all the information required by a user. In addition to the BHL Portal, all the literature will be accessible through Europeana, the portal of the
472
European Digital Library (www.europeana.eu).
Europeana brings together cultural and scientific material from Europe’s libraries, museums, archives and audiovisual archives. BHL-Europe contributed already the first substantial collection of science material to Europeana that is available with the Rhine release of Europeana since 2010.
P0399 – ePoster
Overexpression of AtCHX24, a member of the cation/H + exchangers, accelerates leaf senescence in
Arabidopsis thaliana
Kim, Y-M 1 , Kim, JH 2 , Lee, D-J 3 , Woo, HR 1
1 Dept of Biological Science, Chungnam National
University, Daejeon, Republic of Korea; 2 Division of
Molecular and Life Sciences, Pohang University of
Science and Technology, Pohang, Kyungbuk, Republic of
3 Korea; Graduate School of Analytical Science and
Technology, Chungnam National University, Daejeon,
Republic of Korea
Leaf senescence, the final stage of leaf development, occurs in an age-dependent manner but can be finely regulated by other developmental and environmental factors. Despite the discovery of many genes involved in leaf senescence, the molecular genetic mechanisms of leaf senescence are still unclear. In this study, an activation-tagging based suppressor screen was performed to identify Arabidopsis genes that could suppress the delayed leaf senescence phenotypes of oresara9-1 (ore9-1) when overexpressed. The suppressor1 of ore9 dominant (sor1-D) was caused by the overexpression of AtCHX24, a putative cation/H + exchanger. The sor1-D mutation suppressed the phenotypes of ore9 in age-dependent and dark-induced senescence. Furthermore, the sor1-D mutation restored the delayed senescence phenotypes of ore1 and ore3.
Overexpression of AtCHX24 results in accelerated leaf senescence and suggests that AtCHX24 plays an important role in regulating leaf senescence.
P0400 – ePoster
ABA-GAM 'molecular identification of gametophytes of bryophytes and ferns, and their potential use as bioindicators' (ABA – acceleration of biodiversity asessment)
Lehnert, M 1 , Quandt, D 2 , Nebel, M 2
1 State Museum For Natural History Stuttgart, Germany;
2 Nees Institute For Plant Biodiversity, Bonn, Germany
ABA Ecuador is a research package initiated by the
German Research Foundation and aims to improve existing technologies, and to develop new tools that permit fast and precise assessment of alpha-biodiversity.
Methods that are developed and used in this package of cooperating projects focus on DNA-taxonomy and include new workflows applicable to complex habitats, experiments with new sequencing technologies suitable for mass sequencing, and comparisons of tools required for data analyses. These workflows and methods will be applied to the species-rich tropical mountain forest in

Ecuador, where traditional approaches to quantify alphabiodiversity have only been applied in selected groups due to lacking financial and human resources. The main methodological focus lies on the acceleration of
• biodiversity inventorying,
• species discovery and species description,
• reliable re-identification of known species by nonspecialists. The main ecological tasks are
• to correlate biodiversity patterns with biotic and abiotic habitat parameters,
• to capture baseline data needed for analyses of climate change effects,
• to test hypotheses about ongoing changes of fauna and flora.
The project ABA-GAM 'Molecular identification of gametophytes of bryophytes and ferns, and their potential use as bioindicators' creates an inventory the diversity of free-living gametophytes of liverworts and ferns in southern Ecuador. By comparing gene sequences of sporophytes (or sporophyte bearing gametophytes) and sterile gametophytes along the altitudinal transect, we will fathom the real altitudinal range of the species.
During the period of the project, selected sterile gametophytes colonies outside the optimum zone of the species will be screened annually for formation of sporophytes, which will be aligned with fluctuations in the climate, e.g., temperature peaks during El Niño seasons.
P0401 – ePoster
GrAToL – assembling the green algal Tree of Life
4
1
Lewis, LA
Hall, J 4
1 , Karol, K
Garden, USA; 3
2 , Lopez-Bautista, J
, Delwiche, C 5 , Lewis, P 1
University of Connecticut, USA; 2
3 , McCourt, R 4 ,
New York Botanical
University of Alabama Tuscaloosa, USA;
Academy of Natural Sciences Philadelphia, USA;
5 University of Maryland College Park, USA
Embryophytes inherited their plastids, mitochondria, and basic genetic toolkits from their green algal ancestors.
Elucidation of the evolutionary basis for many land plant traits thus depends on an accurate phylogeny of green algae. Previous morphological and molecular studies have revealed two major clades of green algae (one including embryophytes), but the relationships among major lineages within the green algae still are poorly known. Moreover, within the major groups, relationships and monophyly of many traditionally defined groups are poorly resolved. The current state of phylogenetic knowledge of the group makes green algae one of the most important yet understudied branches on the eukaryotic tree of life. The GrAToL project will assemble a Tree of Life for the green algae, a diverse group of photosynthetic organisms that play important ecological roles in terrestrial and aquatic habitats, from extremely dry deserts to hypersaline ponds, to coral reefs, from photoautotrophic to colorless forms, and as symbiotic photobionts with lichens and animals. A minimum estimate of diversity is just over 14,000 species, with members distributed on all continents and across virtually every habitat type on Earth. The GrAToL project includes over 25 participating scientists, collaborators, and trainees, who contribute taxonomic expertise on the different branches of the green algal tree of life, as well as extensive knowledge of data collection approaches, data analysis, and bioinformatics. We are using an approach that is a hybrid of PCR-based multigene sequencing, morphology and next-generation sequencing methods. The team will assemble a green algal tree of life by sequencing 10 genes for 425 taxa, sampling orthologs from the nucleus, mitochondria, and plastid genomes for phylogenetic analysis. A global analysis will be used to make a reclassification for all green algae. Next-generation sequencing is being for a subset of 16 taxa to study deep phylogenetic relationships in the green algae, identify new target genes for detailed analysis of particular groups, and examine organellar genome evolution in the green algae. The overall goal of this project is to prepare a new classification for all major branches of green algae, in addition to an analysis involving sister groups (red algae, embryophytes). This project will seek to integrate data with other AToL projects (red algae, heterokonts, liverworts and other embryophytes) to identify core genes for broad phylogenetic analyses. A public web site for GRAToL (www.gratol.org) is being developed that includes a library of images, resulting phylogenetic trees, and other resources. We will disseminate new phylogenetic analysis methods through free, open-source software.
P0402 – ePoster
Integrating RNA-seq into plant genome annotation system: a case study of the cucumber genome
1
Lin, K
China;
China
1 , Li, Z 1 , Yan, P 1 , Liao, W 1 , Zhang, Z 2 , Huang, S 2
College of Life Sciences, Beijing Normal University,
2 Institute of Vegetables and Flowers, CAAS,
As more and more genomes recently sequenced, genome annotation plays an increasingly important role in bridging the gap between genomic sequences and biology of organisms. Gene prediction is a key work in genome annotation, which combines various resources to compute consensus gene structures. With the help of the massively parallel cDNA sequencing (RNA-Seq) technology, one can obtain reasonable coverage of a transcriptome resource more efficiently at a lower cost.
RNA-Seq has the potentials to annotate or reannotate less studied eukaryotic genomes for it is highly informative.
Here, we present a pipeline based on EVidenceModeler
(EVM) and incorporated two independent RNA-Seq analysis procedures. We applied this pipeline to the reassembled cucumber genome and compared difference between the genes predicted with and without RNA-Seq reads.
P0403 – ePoster
The mesopolyploid history of Australian and New
Zealand crucifers
Mandakova, T 1
Lysak, M 1
, Joly, S 2 , Mummenhoff, K 3 , Heenan, P 4 ,
1
2
CEITEC, Masaryk Unversity, Brno, Czech Republic;
Université de Montréal and Montreal Botanical Garden,
Montreal, Canada; 3 Universität Osnabrück, Osnabrück,
473

Germany; 4 Allan Herbarium, Landcare Research,
Lincoln, New Zealand
Whole-genome duplication (WGD) events, cyclically increasing the genetic diversity of angiosperm plants, are at different rates followed by genome fractionation toward diploid-like genomes. During recent years, several bona fide diploid species were shown to be ancient polyploids (Soltis et al. 2009). The incidence and age of WGD events, as well as the extent of subsequent genome diploidization vary widely between angiosperm phyla. Due to the feasibility of comparative chromosome painting (CCP) in the Brassicaceae, we could reconstruct the fate of a mesopolyploid WGD event in 'diploid'
Australian and New Zealand crucifer species
(Mandáková et al. 2010a, b). CCP analysis showed that genomes of endemic Australian species ( Stenopetalum nutans, S. lineare, and Ballantinia antipoda with n=4, 5, and 6, respectively) descended from the eight ancestral chromosomes (AK1-AK8) through an allopolyploid
WGD. Nuclear and maternal gene phylogenies corroborated the hybrid origin of the mesotetraploid ancestor and suggest that the WGD occurred c. 6 to 9 million years ago (mya). Similarly, the ancestral genome of ten New Zealand Pachycladon species (all with n=10) originated through a mesopolyploid event involving two genomes with the eight ancestral chromosomes, c. 1 to 2 mya. Both, presumably independent, WGD events were followed by taxon-specific reductions of chromosome number, with less extensive inter-genomic reshuffling in the evolutionary younger Pachycladon ancestor. Intergenomic rearrangements and chromosome 'fusions' were mediated by inversions, whole-arm translocations, and by end-to-end translocations associated with centromere inactivation and/or loss. Our data underline the importance of multiple whole-genome duplication events in the angiosperm genome evolution, elucidate diploidization mechanisms transforming polyploid genomes into 'diploid' ones, and demonstrate that chromosome number per se is not a reliable indicator of ploidy level.
P0404 – ePoster
Development of polymorphic microsatellite markers for
Pimenta pseudocaryophyllus
(Gomes) Landrum,
Myrtaceae
, Sebastião, I 1 , Silveira, LED 1 , Mori, Morgante, PG 1
1
GM 2
UNESP, Registro, SP, Brazil; 2 UNICAMP, Campinas,
SP, Brazil
Pimenta pseudocaryophyllus (Gomes) Landrum, popularly known as cataia in Brazil, belongs to the
Myrtaceae, an important taxon in the Brazilian flora being one of the dominant woody families in the Mata
Atlântica rainforest. This species is the only one from the genus found in southeastern and southern South
America. In the Vale do Ribeira (São Paulo, Brazil), this species is widely consumed for medicinal purposes as a tea in fighting colds, flu and, fatigue and as a diuretic; it is also used for inhalations and massages. Besides, currently, an increasing commercialization of its leaves as a flavoring agent of alcoholic drinks is being observed.
Despite of this great variety of human uses for this plant,
474 little is known about the consequences of this anthropogenic pressure or even about its biology. In order to perform studies of genetic diversity and population structure of natural populations of cataia, a microsatellite-enriched library was constructed and 16 pairs of primers were designed to develop Simple
Sequence Repeats (SSR) markers. It was then carried out a preliminary analysis of these loci using five individuals from different populations of P. pseudocaryophyllus . Of the 16 loci tested, 15 allowed the amplification of PCR products in the range of expected size when analyzed on
3% agarose gel. The genotyping of these 15 loci in 6% denaturing polyacrylamide gels revealed the presence of
11 polymorphic loci with typical banding pattern of diploid species. We found 2–6 alleles in these five individuals and Polymorphism Information Content
(PIC) values ranged from 0.2688 to 0.7942. Two loci showed a significant deviation from the Hardy-Weinberg
Equilibrium (HWE; p <0.05). Currently, these 11 loci are being analyzed for a total of 35 individuals from different populations in order to validate the results. These molecular markers will be valuable tools in order to understand more about the biology of P. pseudocaryophyllus and detect ongoing consequences of its use, in a conservation genetics approach. Financial support: FAPESP – Regular Research Award
(2009/03595-0) and Undergraduate Research Scholarship
(2009/15964-0).
P0405 – ePoster
Genetic differentiation in populations of
Pyricularia grisea
(Cooke) Sacc obtained from various hosts, based on detection of VCGs and rep-PCR
Motallebi, P 1 , Nikkhah, M 1 , Okhovvat, M 1
1 Dept of Plant Sciences, School of Biology, College of
Science, University of Tehran, Iran
Pyricularia grisea is the causal agent of blast disease on rice and leaf spot on some Poaceae weeds in Iran and other regions of the world. In this study, ninety six isolates were recovered from rice and some weeds, including 40 isolates of rice, 48 isolates of weeds, counting Digitaria sanguinalis (crabgrass), Setaria italica (foxtail millet), Echinochloa crus-galli (barnyard millet), and some unknown weeds and 8 standard mating type tester isolates which were isolated during 1997–
2006 were examined for characterization of genetic diversity. Complementation tests of nit mutants were done in all possibilities on minimal medium (MM). As a result, four VCGs for isolates from rice and three VCGs for weeds were separately identified and no heterokaryon formation was observed between these two populations and any nit mutants obtained from standard mating type tester. For identification of clonal lineages in populations of the fungus, using two primers based on ERIC and
BOX regions. Twelve clonal lineages, A to L, were identified. 'A' clonal lineage was the dominant among others.'E'clonal including isolates from crabgrass was separated with 42% genetic similarity. The results were confirmed each other in comparing of VCG and rep-
PCR and no association was seen among isolates obtained from different hosts either in VCGs or rep-PCR.
The only ambiguity is about foxtail millet which is associated as the same VCGs with isolates from

crabgrass but was identified with more than 65% genetic similarity with isolates from rice. The standard mating type tester was as well branded to 4 separate lineages, H to L. The correlation between VCGs and clonal lineages revealed low genetic diversity among all the isolates and far relationship between isolates from rice and crabgrass, whereas the possibility of speciation.
P0406 – ePoster
Studies on B-chromosomes in conifers
Muratova, E 1
1 V.N. Sukachev Institute of Foresy, Russian Academy of
Sciences, Siberian Branch, Russia
B-chromosomes are found in animals, musci, gymnosperm and angiosperm plants. At present about 30 species with B-chromosomes are known among conifers.
The paper deals with the results of B-chromosome studies in representatives of Pinaceae family. Among investigated by author species B-chromosomes are found in two Larix (larch) species and nine Picea (spruce) species. For karyological investigations seeds of different coniferous species (Pinaceae family) from many provenances were used. The materials were prepared and analyzed according to generally accepted techniques for coniferous plants with some modifications. The germinating seeds were pretreated in 0.5% colchicine solution for 6-8 hours, fixed in 3:1 ethanol : acetic acid mixture and stained with acetohematoxylin. Root tip meristem cells were used for study, and slides were prepared using the improved squash technique. Species from the genera Picea and Larix are the stable diploids and contain 24 somatic or A-chromosomes (2n=24). On morphological type in Picea 8 pairs of A-chromosomes are long metacentrics and 4 pairs of A-chromosomes are short meta- or submetacentrics. Karyotypes of some species include B-chromosomes. In P. schrenkiana, P. jezoensis, P. pungens, P.
x fennica and P. breweriana one B-chromosome occurs (2n=24+1B), in P. koyamae and P. engelmannii 1–2 (2n=24+1–2B), in P. ajanensis and P. meyeri 1–3 (2n=24+1–3B), in P. obovata 1–4
(2n=24+1–4B), in P. glehnii 1–5 B-chromosomes
(2n=24+1–5B). It was the first study case occurrence of
B-chromosomes in four species – P. breweriana, P. pungens, P. koyamae and P. schrenkiana.
At present Bchromosomes are found in 19 Picea species. Larix karyotype includes 6 pairs of long metacentric chromosomes and 6 pairs of short submetacentric ones.
B-chromosomes are found in two Larix species: L. gmelinii and L. sibirica . Occurrence of B-chromosome in
L. gmelinii was the first study case for the genus Larix .
Sizes of A-chromosomes Picea and Larix are from 9 to
15
µ m. Length of B-chromosomes of these species composes 25–30 % from the A-chromosomes one (4–6
µ m). On their morphology B-chromosomes of Picea species can be metacentric (B1 type) and submetacentric
B2 type. In P. schrenkiana, P. koyamae and P. breweriana the first type of B-chromosomes occurs only.
In P. pungens the second type of B-chromosomes occurs only. P. obovata, P. ajanensis, Larix gmelinii , L. sibirica have the both type of B-chromosomes. It is supposed that submetacentric B-chromosomes originated in the result of pericentric inversion of metacentric ones. Other variants of B-chromosomes were found in P. glehnii . In this species five morphological types of B-chromosomes were found: large metacentric, two short meta- and submetacentric, one small metacentric, and very small submetacentric. In the opinion of some authors presence of B-chromosomes can be connected with unfavourable ecological factors (Moir, Fox 1977; Teoh, Rees 1977;
Jones, Rees 1982; Broka 1990; Muratova et al. 2001,
2002 and others). Effects of B-chromosomes can have adaptive character. The results obtained allow us to consider that this system is a general phenomenon and Bchromosomes are of importance for populations and species and possibly play a role for their adaptation.
P0407 – ePoster
Development and characterization of microsatellite markers on Pistachio (
Pistacia vera
)
Sola Campoy, PJ
D 2
1
, Gijón López, MC 2
Rejón, C 1
, Madsen, L 1 , Robles, F 1
, Reche, M 3
, de la Herrán, R 1 ,
, Pérez López,
, Benavides, C 3 , Ruiz
Navajas-Pérez, R 1
1 Departamento de Genética, Facultad de Ciencias,
Universidad de Granada, Spain; 2
Chaparrillo, Ciudad Real, Spain;
Centro Agrario El
3 Centro Agrario
IFAPA, Granada, Spain
Pistacia vera L. (Anacardiaceae) is an economically important species as the main source of pistachio nuts.
The application of genetic tools for the improvement of its commercial culture is crucial. In this sense, a linkage map based on molecular markers of this species will facilitate future breeding programs. The linkage maps are useful in important aspects such as sex-determination or marker-assisted selection for desirable traits issues
(disease-resistance, growth rate, productivity…). The characterization of DNA markers is the first step towards the development of these maps. In this sense, we have constructed microsatellite-enriched libraries in pistachio by using the hybrid capture method. The libraries were made from genomic DNA samples and were enriched in di- and trinucleotides. Different libraries for males –
Peter cultivar – and females – Kerman cultivar – were constructed to explore the possible association of these markers with the sex-determining loci. We also examined the utility of conserved microsatellite markers in other important Pistacia species (i.e. P. lentiscus or P. terebinthus). We believe that these data will contribute to a better knowledge of this group of species.
P0408 – ePoster
Characterization of repetitive sequences on Pistachio
(
Pistacia vera
)
Sola Campoy,PJ
1
Villaseñor, J 2
Herrán, R 1
1 , Madero Pérez, J 1
, Couceiro López, FJ
, Navajas-Pérez, R 1
2
, Robles, F 1 , Guerrero
, Ruiz Rejón, C 1 , de la
Departamento de Genética, Facultad de Ciencias,
Universidad de Granada, Spain;
Chaparrillo, Ciudad Real, Spain
2 Centro Agrario El
The genus Pistacia L. (Anacardiaceae) includes around a dozen of species. Some of them have economical and ecological importance, highlighting the species Pistacia vera – the source of pistachio nuts – or Pistacia
475

terebinthus and Pistacia lentiscus – important elements of Mediterranean flora. Moreover, all Pistacia species are dioecious. Dioecy is a rare condition that occurs in
~7% of flowering plants genera. For all these reasons, this group of plants represents an interesting model for biological analyses being good candidates to shed light on the way gender is determined in plants. Repetitive
DNA has been proved a useful tool for several evolutionary studies. Thus, it is commonly used as a molecular marker in taxonomic and phylogenetic analyses. Furthermore, its implication on sexchromosome origin and evolution has been reported. In the present work, we aim to isolate different repetitive
DNA sequences from pistachio genome, focusing on satellite-DNA, a significant fraction of the eukaryotic genome. These repeats are tandemly-arrayed sequences.
They are the main constituents of heterochromatin and show a pattern of evolution known as concerted evolution. The characterization of repetitive DNA from
Pistacia species genome will allow us to perform different evolutionary analysis. Their organization, chromosomal location and possible differential accumulation between males and females individuals and their implication on pistachio tree sex-determination will be also discussed. Additionally, these repetitive sequences will be useful DNA markers for early identification of gender in the pistachio cultivars.
P0409 – ePoster
Molecular analysis of adaptive traits associated with flooding stress tolerance in the genus
Oryza
Niroula, RK 1
1 Biotechnology Division, Nepal
Crop tolerance to flooding is an important agronomic trait. Although rice ( Oryza sativa ) is considered a floodtolerant crop, only some cultivars display a real tolerance to prolonged submergence, which is largely attributed to the presence of the Sub1A gene. Secondary Oryza gene pools have the potential to unveil new adaptive mechanisms and shed light on the evolution of submergence tolerance traits. In this study, we screened
109 Oryza genotypes for flooding tolerance. O. nivara and O. rufipogon accessions showed a wide range of submergence responses. The tolerance-related Sub1A-1 and the intolerance-related Sub1A-2 alleles were found in tolerant and sensitive accessions respectively. This suggests that the presence of the Sub1A gene is the result of evolutionary pressure on the Oryza species in flooding-prone areas. Flooding tolerant O. rhizomatis
(IRGC-103421) and O. eichingeri (IRGC-101429) genotypes were also identified. Sub1A-1 was absent in these two species, which possess Sub1 orthologues with a high degree of similarity to O. sativa Sub1C. The coding sequence of these Sub1C-like genes revealed the presence of truncated versions of the transcript in correspondence to the ERF domain. Although Sub1A-1 was absent from these species, the expression patterns of submergence-induced genes were similar to tolerant O. sativa genotypes containing Sub1A-1. The results indicated that Sub1A-1 is not essential to confer submergence tolerance in rice relatives with the C genome that were analyzed.
P0411 – ePoster
Relationships between three flowering cherry cultivars bearing green flowers
Oyama, E 1 , Uematsu, C 1 , Oohara, T 2
1 Botanical Gardens of Osaka City University, Japan;
2 Botanic Gardens of Toyama, Japan
Flowering cherry is a well-known flower in Japan. It is also very popular as an ornamental plant all over the world. We have investigated three flowering cherry cultivars bearing green flowers: Prunus ‘Grandiflora’ bearing yellow-green flowers, Prunus ‘Gioiko’ bearing variegated flowers with light green sectors and Prunus
‘Shin-nishiki’ bearing variegated flowers with deep green sectors. One ‘Shin-nishiki’ tree has been conserved in the
Botanical Garden of Osaka City University. There are very few records about this cultivar, but it would appear that no other trees with the name of ‘Shin-nishiki’ have survived. Recently there has been confusion concerning nursery trees of ‘Gioiko’. Two different types have been called ‘Gioiko’. One is the original ‘Gioiko’, and another is ‘Shin-nishiki like Gioiko’. The latter is believed to have arisen during proliferation. In this study we tried to clarify the characteristics of the three original cultivars and reveal the relationships among cultivars.
All three cultivars are double-flowered. The diameter of
‘Shin-nishiki’ flowers was on average 2.8 cm. This was smaller than that of ‘Grandiflora’ and ‘Gioiko’ which both had flowers bigger than 4.0 cm in diameter. The length and width of the petals were also smaller in ‘Shinnishiki’ compared to the other two cultivars. ‘Shinnishiki’ had intermediate petal-stamen floral organs. The shape of the calyx tube is one of the important characteristics distinguishing different cultivars of flowering cherry. ‘Shin-nishiki’ had a funnel-shaped calyx tube, whereas ‘Grandiflora’ and ‘Gioiko’ had bellshaped ones. Flowers of ‘Grandiflora’ and ‘Gioiko’ possessed similar morphological traits, whereas the flower colour of these cultivars differed as described above. All three cultivars have stomata on their petal surfaces as shown by using a KEYENCE digital microscope (KEYENCE, Japan) and SEM Miniscope
(HITACHI, Japan). More stomata were observed on the abaxial side of petals, and the number of stomata was higher in the green sectors than in the white background area. ‘Shin-nishiki’ had high stomatal density of ca
20/mm2, whereas ‘Grandiflora’ and ‘Gioiko’ had only 7–
8 stomata /mm2. In contrast, almost no stomata were observed in other flowering cherry cultivars bearing pink flowers used as a control. The total chlorophyll content of petals of ‘Shin-nishiki’ was 0.32mg/gFW, which was fourfold higher than in ‘Grandiflora’ or ‘Gioiko’. This result agreed well with the differences in flower colour appearance between the three cultivars.
Based on the morphological characteristics of the flowers, ‘Shin-nishiki’ could be distinguished easily and clearly from the other two green cherry cultivars. Flower characteristics of ‘Shin-nishiki like Gioiko’ were almost same as those of ‘Shin-nishiki’. Molecular analysis using
SSR markers is now ongoing so that genetic relationships among three cultivars (which include four types) can be defined. Stomatal distribution and total chlorophyll
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content in ‘Shin-nishiki’ suggested reversion in green areas of variegated petals. The labeling of two varieties as ‘Gioiko’ might suggest the mechanism by which
‘Shin-nishiki’ arose.
P0412 – ePoster
1
Apiaceae organellar genomes
Peery, R 1 , Downie, SR 1 , Jansen, RK 2 , Raubeson, LA 3
2
University of Illinois At Urbana-Champaign, USA;
University of Texas Austin, USA;
University, Ellensburg, USA
3 Central Washington
Apiaceae is a very diverse, speciose, and geographically widespread family. Many members are economically important such as dill, fennel, anise, and parsley.
Apiaceae is one of the few families known to have a dynamic inverted repeat in the plastid genome. Not only are expansions and contractions of the inverted repeat common, there is also an insertion in the sequence between the end of the inverted repeat and the start of the gene trn H. However, little is known at the sequence level about how extensive these changes are, if the changes have occurred more than once, and the possible mechanisms for these changes. In addition to seven completely sequenced plastid genomes (including the previously sequenced Daucus chloroplast genome), we have sequenced the inverted repeat/large single copy junctions of 30 taxa from different genera, with partial sequences for 70 additional taxa (out of 200 planned).
Thus far our data does not support the hypothesis that boundary shifts are due to mononucleotide repeats, as has been hypothesized in other taxa. In fact, the same repeat motif is present near the gene rps19 even if the IR boundary is in another location. Mononucleotide repeats are common and can be expected to occur every 100bp of intergenic sequence. Our data also show that extreme contractions of the inverted repeat, such as those seen in
Coriandrum sativa and Bifora radians (tribe
Coriandreae), have occurred differently, even though the extent of the inverted repeat is identical in mapping studies. Also, there appears to be unique interactions
(potentially intracellular gene transfer) between the mitochondrial and plastid genomes in this family that have been referred to twice in the literature but never empirically investigated. This interaction may be a case of intracellular gene transfer from the mitochondrial genome to the plastid. Currently we are isolating mitochondrial DNA to use as a template for gene walking to determine the extent of sequence similarity between the mitochondrial DNA and the unique insert found near the inverted repeat boundary in several clades. We are also looking further into the extent of this novel sequence within Apiaceae chloroplast genomes to determine when the sequence entered the Apiales, identifying the source of the sequence, and ascertaining whether or not the inverted repeat and mitochondrial sequence data can be used successfully as a phylogenetic marker in the Apiales.
P0414 – ePoster
Genetic improvement of Urdbean (
Vigna mungo
L.
Hepper) using induced mutations
1
Sagade, A 1
Jijamata College of Science and Arts, Bhende,
Maharashtra State, India
The main objective of urdbean breeding programmes has been to develop improved urdbean cultivars. In an attempt to induce genetic variability for improvement of locally popular urdbean cultivar TAU-1, we employed three well known mutagens, ethyl methane sulphonate
(EMS), methyl methane sulphonate (MMS) and gamma radiation (GR). The objective was to provide genetic variability in the yield contributing traits that can be exploited for a genetic improvement of urdbean. Seeds of urdbean cultivar TAU-1 were treated with three different concentrations / dose of EMS (0.02, 0.03 and 0.04M),
MMS (0.0025, 0.005 and 0.01M) and gamma radiations
(30, 40 and 50 KR). In M1 generation no dominant mutation were observed, different mutants were screened and isolated in M2 generation such as chlorophyll mutations (chlorine, xantha and viridis); leaf mutations
(curly leaf and thick leaf); pod mutations (long pod, short pod, hairy pod, narrow elongated pod, flat pod and large pod); seed mutations (small seed, bold seed and wrinkled seed); morphological mutations (sterile, tall, dwarf, viney, spreading, early and high yielding). True breeding mutant lines in M3 generation differed considerably in their quantitative traits from the parent cultivar. The early mutant lines matured 10-15 days earlier than the parent variety. Tall mutant were tall over the control. Spreading, high yielding and tall mutant showed significantly increase in the number of pods and seeds per plant.
Mutagenic treatments also caused changes in pod and seed size. Considerable genotypic variation was observed with regards to the number of pods and seed yield per plant. As a result of mutagenic treatments, genetic variation was induced in mutants with respect to different quantitative characters. Induced mutant lines showed both positive and negative increase in the quantitative traits. Variation was also observed for total protein, chlorophyll and carbohydrates content of mutants. Key words: Urdbean, EMS, MMS, Gamma radiation,
Mutation
P0416 – ePoster
Cyto-nuclear interactions in
Tragopogon
(Asteraceae)
Allopolyploids: insight from rbc
L and rbc
S
Sehrish, T
Tate, JA 4
1 , Symonds, VV 1 , Soltis, DE 2 , Soltis, PS 3 ,
1
4
Institute of Molecular BioSciences, Massey University,
Palmerston North, New Zealand; 2 Dept of Biology,
University of Florida, Gainesville, USA; 3 Florida
Museum of Natural History, University of Florida, USA;
Institute of Molecular BioSciences, Massey University,
New Zealand
Gene products from organellar genomes (chloroplast and mitochondria) can interact with nuclear gene products in a variety of pathways. Most protein complexes consisting of multiple subunits have proteins encoded by both nuclear and organellar genomes. Ribulose-1, 5bisphosphate carboxylase/oxygenase (RuBisCO) is involved in carbon dioxide fixation in the Calvin cycle and has two subunits: a large subunit ( rbc L) encoded by
477

the chloroplast genome and a small subunit (rbcS) that is encoded by the nuclear genome. How these subunits interact in hybrids and allopolyploids is unknown. In order to explore the inheritance and expression of both maternal and paternal homeologs of the nuclear-encoded rbc S, and to determine which of the nuclear homeologs of rbc S is interacting with the chloroplast-encoded subunit rbc L, rbc S was amplified from three diploid species of Tragopogon ( T. dubius, T. pratensis and T. porrifoilius ), which are the parents of the allopolyploids
T. mirus and T. miscellus . Sequence analysis of rbc S from the diploid parents showed the presence of seven single nucleotide polymorphisms (SNPs). Based on this variation among the parents, progenitor-specific markers are being designed, which will be used to amplify rbcS homeologs from reciprocally generated synthetic polyploid lines of T. mirus and T. miscellus . We expect that both parental copies of rbc S will be present in the genomes of the synthetic polyploids, but that the maternal homeolog of rbc S will be expressed and will interact with rbcL from that same progenitor.
P0419 – ePoster
Fine mapping of a major QTL, qFLL62, controlling flag leaf length and yield traits in rice
Shen, B 1 , Zhuang, J-Y 2
1 College of Life & Environmental Sciences, Hangzhou
Normal University, China; 2 China National Rice
Research Institute, China
Population derived from a residual heterozygous line of the indica rice cross Zhenshan 97/Milyang 46 was used for the fine mapping of a quantitative trait locus (QTL), qFLL6.2, controlling flag leaf length (FLL) and yield traits in rice. Four sets of near isogenic lines (NILs) were developed, of which each consisted of 10 maternal homozygotes, 10 paternal homozygotes and 20 heterozygous differing in a portion of the 1.19-Mb interval RM3414-RM6917 on the short arm of rice chromosome 6. With analysis of phenotypic differences among three genotypic groups in each NIL set, qFLL6.2 was delimitated into a 62.1-kb region flanked by SSR marker RM3414 and STS marker Si2944. It explained
52.73% of the phenotypic variance, and the Zhenshan 97 allele increased FLL by 2.40 cm. The QTL effect was steadily detected in two F2 populations segregating qFLL6.2 in an isogenic genetic background. Based on data collected from homozygous lines of three of the NIL sets, qFLL6.2 was shown to have major effects on all the three yield traits analyzed, including number of spikelets per panicle, number of grains per panicle and grain weight per panicle. On comparison among reports of different groups, the effect of qFLL6.2 was found to be highly consistent across different genetic backgrounds and environments, providing a good candidate for mapbased cloning and investigating the source-sink relationship in rice.
P0420 – ePoster
Harnessing next-generation sequencing technology for DNA marker development
Shepherd, L 1 , Atherton, R 1
McComish, B 1
Lockhart, P 1
, Cox, S
, McLenachan, T 1
1 , Holland, B 2
, Monaghan, J 1
,
,
2
1 Massey University, Palmerston North, New Zealand;
University of Tasmania, Hobart, Australia
Next-generation sequencing technology has enabled large quantities of DNA sequence data to be obtained quickly and cheaply. We are evaluating a number of novel methods using the Illumina GAII to obtain and screen variable molecular markers for answering molecular ecology and evolution questions. This includes sequencing both whole and partial chloroplast genomes in order to develop SNP markers for studying the recent domestication of two native New Zealand plant species.
We have also sequenced random nuclear loci from a genus of New Zealand trees in order to develop a multilocus nuclear dataset for investigating the influence of hybridization on phylogeny reconstruction.
P0423 – ePoster
DArT markers provide a powerful tool for population genetic analysis and phylogeny reconstruction in
Eucalyptus
(Myrtaceae)
Steane, DA 1 , Hudson, C 1
Petroli, CD 3 , Carling, J
Grattapaglia, D 3,4,7
5
, Nicolle, D 2
, Kilian, A
, Vaillancourt, RE 1
, Sansaloni, CP 3
5 , Myburg, AA
,
6 ,
1 University of Tasmania, Hobart, Australia;
Creek Arboretum, Australia;
2 Currency
3 EMBRAPA Genetic
6
Resources and Biotechnology, Brazil;
Brasília, Brazil;
4 Universidade de
5 Diversity Arrays Technology Pty Ltd;
University of Pretoria, South Africa;
Católica de Brasília, Brazil
7 Universidade
A set of over 8000 Diversity Arrays Technology (DArT) markers developed for genomic studies of Eucalyptus was tested for its utility in population and phylogenetic studies. Small-scale population studies of E. camaldulensis, E. cladocalyx, E. globulus, E. grandis, E. nitens , and E. urophylla (from subgenus Symphyomyrtus ) and E. pilularis (subgenus Eucalyptus ) demonstrated the potential of DArT markers to differentiate species, identify interspecific hybrids and resolve biogeographic disjunctions within species. Phylogenetic reconstruction of 94 eucalypt species using the DArT markers was largely congruent with traditional taxonomy and ITSbased phylogenies, but provided greater resolution. DArT offers high-throughput genome-wide genotyping suitable for population genetic, phylogenetic and evolutionary studies across the full range of Eucalyptus species.
P0424 – ePoster
Photosynthetic response to heat stress and geographic range of two C
3
grass species in Japan
1
Sugiyama, S-I 1
Hirosaki University, Japan
Orchardgrass ( Dactylis glomerata L.) and tall fescue
( Festuca arundinacea Schreb), C
3
grass introduced into
Japan about 120 years ago as a forage species, have
478

established naturalized populations in disturbed areas in northern Japan through natural colonization from seeds.
The two species reveal different distributional pattern.
Naturalized populations of D. glomerata show decline in their relative abundance along a north–south gradient, whilst those of F. arundinacea do not show a large decline. These distributional patterns suggest that the two species differ in heat tolerance. This study was conducted to examine (1) whether photosynthetic responses to high temperature are related to different pattern of geographic distribution between the two species, and (2) if any, which photosynthetic parameters are involved into the abundance decline in D. glomerata . Plants of the two species were grown in a controlled growth chamber with day/night temperatures of 23/16°C, a 16-h photoperiod
(4:00 to 20:00 h) with photon flux of 250 µmol m –2 s –1 for 40 days. The plants were exposed to 30ºC for 3 days for acclimation and then exposed to 7 different temperature regimes (28°C, 30°C, 32°C, 34°C, 36°C and
40°C). Plants were exposed to each temperature durig three days and A/Ci curve was measured at the third day of each temperature to estimate Jmax (RuBP regeneration capacity) and Vcmax (maximum rate of
Rubisco). To measure tolerance to long-term heat stress, the maximum PSII activity (chlorophyll fluorescence,
Fv/Fm) was also monitored at 36/30ºC (day/night) during
40 days. (1) Photosynthetic rate (Pr) largely declined above 30°C. However, there were no apparent differences in the decline pattern between the two species. (2) Jmax largely declined above 30°C, but little changes in Vcmax. These results showed that RuBP regeneration is more influenced by high temperature than does Rubisco activity. However, there were no significant differences in the two parameters between the two species, indicating two species do not differ in responses of RuBP regeneration and Rubisco activities to high temperature. (3). The two species did not significantly differ in Fv/Fm until 30 days after the exposure of 36°C, but D. glomerata showed a large decline at 40 days of exposure. These results demonstrate that D. glomerata , which show abundance decline along a north–south gradient, has low ability to tolerate prolonged heat stress than F. arundinacea .
P0425 – ePoster
Investigating the roles of barley CesA genes in cell wall biosynthesis
Tan, AH-T 1, 2 , Burton, R 2 , Fincher, G 2
1 Australian Centre For Plant Functional Genomics,
2 Australia; ARC Centre of Excellence for Plant Cell
Walls, School of Agriculture, Food and Wine, Waite
Campus, University of Adelaide, Urrbrae, Australia
Barley contains at least eight cellulose synthase (CesA) genes that have been transcript -profiled in a range of tissues (Burton et al. 2004). CesA1, CesA2 and CesA6 have higher transcript levels in tissues more likely to be undergoing the synthesis of primary cell walls, while
CesA4, CesA5/7 and CesA8 are more likely to be involved in the synthesis of secondary cell walls (Burton et al. 2004). Growing and expanding cells mainly consist of primary walls and secondary cell walls are usually deposited upon cessation of cell growth (McNeil et al.
1984). In this study, a gain-of-function approach was used to study the roles of the CesA genes in barley cell wall biosynthesis. Five sets of transformed barley plants transgenic for constructs carrying the CesA1, CesA2,
CesA4, CesA6 or CesA8 genes driven by the constitutive
35S promoter were analysed for transcript levels, phenotype and tissue morphology. Preliminary results showed that transcript levels of the CesA transgenes were lower than expected even though they were driven by a strong constitutive promoter. The transgenic lines transformed with the secondary cell wall CesA4 exhibited a phenotype where xylem cells in the vascular tissue appear to be irregular in shape and are partially collapsed. This resembles the irx mutants described in
Arabidopsis (Turner and Somerville 1997). A 'brittle node' phenotype was also observed in barley transgenic for 35S:HvCesA8 which also may be affecting secondary cell wall synthesis. Further studies will include stem strength measurements and cellulose content assays of the transgenic barley stems.
P0426 – ePoster
Mechanism of flower colour variegation in
Camellia bearing pigmented flowers and variegated ones within a single tree
Taniguchi, K 1 , Uematsu, C 1 , Arakawa, O 2
1 Botanical Gardens of Osaka City University, Katanoshi, Japan; 2 Hirosaki University, Hirosaki-shi, Japan
Camellia japonica is famous as an ornamental flower used in the traditional Japanese tea ceremony. Many cultivars have been grown all over the world for ornamental and also for oil production. Some camellia cultivars bear fully pigmented flowers, variegated flowers and occasionally white flowers on a single tree.
This phenotype seems to have arisen through bud mutation. Even though bud mutation is advantageous for woody plants to produce genetic diversity within an individual, the mechanism of mutation remains unclear in many cases. In this study we investigated the mechanisms of flower colour variegation using two cultivars ‘Kyotosano’ and ‘Gisetsu’ both conserved in the Botanical Gardens of Osaka City University.
‘Kyotosano’ bears red flowers and variegated flowers with deep red sectors on a white background, whereas
‘Gisetsu’ bears pale pink flowers, variegated flowers with pale pink sectors on a white background, and completely white flowers. The distribution of pigmented cells was observed by microscope using hand sections of petals. In the case of fully pigmented petals, regardless of whether they were pink or red, only the epidermal layer was pigmented. The inner cell layers of the petals were not pigmented. In contrast, in variegated petals, pigmented cells were observed not only the epidermal layer but also in the inner cell layers. There were no pigmented cells in the white petals. Pigment analysis was performed by HPLC and showed that the major pigment was cyanidin-3-glucoside in both cultivars. Differences in flower colour were due to differences in the content of cyanidin-3-glucoside. Pink petals of ‘Gisetsu’ contained on average 0.31 mg/gFW cyanidin-3-glucoside, whereas red petals of ‘Kyotosano’ contained about 3.0mg/gFW.
Variegated petals included smaller amounts of cyanidin-
3-glucoside compared to the fully pigmented petals in each cultivar. No anthocyanins were detected in the
479

white petals of ‘Gisetsu’. These results indicated that suppression in certain step of anthocyanin biosynthetic pathway might cause the failure in pigment production and result in the white regions of the petals. The expression of genes encoding enzymes involved in the anthocyanin biosynthetic pathway i.e., phenylalanine ammonialyase (PAL), chalcone synthase (CHS), dihydroflavonol-4-reductase (DFR), flavanone-3hydroxylase (F3H) and anthocyanidin synthase (ANS) was investigated by RT-PCR using RNA extracted from petals of young flower buds. After 40 cycles of amplification, PAL, CHS, F3H and DFR genes were expressed equally in both pigmented and variegated petals. However expression of the ANS gene was detected only in fully pigmented petals of both
‘Kyotosano’ and ‘Gisetsu’, and not in variegated petals of either cultivar nor in white petals of ‘Gisetsu’. This result suggested the involvement of the ANS gene in flower colour variegation in these cultivars. The mechanisms regulating ANS expression need to be investigated further.
P0428 – ePoster
Centromere-specific integration of Copia-type LTR retrotransposon in
A. lyrata
, Kawabe, A 3 , Kobayashi, A 1 , Kakutani, Tsukahara, S 1,2
1
T 3
National Institute of Genetics, Japan; 2 Graduate
3
University for Advanced Studies (SOKENDAI), Japan;
Kyoto Sangyo University, Japan
Retrotransposons are major component of plant genome.
In plants, many of retrotransposons show clustered distribution, generating pericentromeric heterochromatin or knob. One possibility for biased distribution is targeted integration of retrotransposons into such region.
However, no mobile retrotransposon with targeted integration has been found in plants. We have recently found mobile retrotransposons in ddm1 (decrease in
DNA methylation 1) mutant, which shows reduced DNA methylation in genome (Tsukahara et al, 2009 Nature).
We have also found that COPIA93 family LTR (long terminal repeats) retrotransposon, which is one of the mobile retrotransposon families in ddm1, showed centromere-specific localization in A. lyrata . Here, we directly show that targeted integration of COPIA93 family retrotransposon in A. lyrata , which we named
Tal1 (Transposon of Arabidopsis lyrata 1), into centromeric repeats in A. thaliana genome. We introduced Tal1 into A. thaliana by Agrobacterium mediated transformation. As a result, Tal1 was transcribed and retrotransposed in transgenic plants.
Most importantly, Tal1 showed strong preference for integration into 178bp centromeric satellite repeats, suggesting that Tal1 has the ability to target centromeric repeats specifically.
P0429 – ePoster
The S-locus genomic region in
Arabidopsis halleri
and
A. lyrata
presents molecular evolution properties analogous to sex chromosomes
Goubet, P 1 , Vekemans, X 1 , Berges, H 2 , Castric, V 1
480
2
1 GEPV, FRE 3268 CNRS, University Lille, France;
CNRGV, IINRA Toulouse, France
Self-incompatibility is a common genetic system preventing self-fertilization in hermaphrodite flowering plants. In homomorphic self-incompatibility, two highly linked genes presenting many different alleles are responsible for the pollen-pistil recognition step necessary to trigger the incompatibility reaction in the presence of self-pollen. In the Brassicaceae family, exhibiting sporophytic self-incompatibility, the pollen gene, SCR, produces a ligand protein, and the pistil gene,
SRK, produces the receptor. Both genes are localized in a region called the S-locus. Because many different allelic specificities have been maintained over millions of years at this S-locus in the Brassicaceae, under the action of strong negative frequency-dependent selection, the Slocus region presents highly divergent haplotypes. In order to study the molecular evolution properties of the
S-locus genomic region in the self-incompatible species
Arabidopsis halleri and A. lyrata , we built genomic libraries for a dozen of S-haplotypes and we sequenced corresponding BAC clones comprising the entire S-locus region. We found strong homologies and collinearity of
ORFs in the flanking regions on both sides of the Slocus, that could be aligned easily against the public A. thaliana and A. lyrata genomic sequences. In striking contrast, the S-locus region in itself showed very little degree of homology among haplotypes, and high variability in size (30-110 kb). In each haplotype, we could find one copy of SCR and one copy of SRK, but the locations and orientations of these genes where highly variable. The phylogenies of the two genes were highly congruent, showing clear evidence of co-evolution of the pollen and pistil determinants of selfincompatibility. In some haplotypes, we found high numbers and densities of transposable elements, of variable size and origin. Altogether, our results show that the S-locus genomic region in self-incompatible species of the Brassicaceae presents many similarities with Ychromosomes in vertebrates or in plants, including an apparently sharp transition from non-recombining regions to highly recombining, pseudo-autosomal regions, represented by the flanking regions on both sides of the S-locus.
P0430 – ePoster
Development of large sets of simple sequence repeat
(SSR) markers for three species of subfamily
Pitcairnioideae (Bromeliaceae) using high-throughput
1
454 pyrosequencing
Wöhrmann, T 1 , Huettel, B 2 , Krapp, F 1 , Weising, K 1
Institute of Biology, University of Kassel, Germany;
2 Max-Planck-Institute for Plant Breeding, Cologne,
Germany
We used the 454 Life Sciences / Roche GS FLX
Titanium platform to generate 25,827, 59,624 and 73,027 reads with an average length of 341 bp from
Deuterocohnia brevifolia, Dyckia marnier-lapostollei and Fosterella rusbyi (all Pitcairnioideae; Bromeliaceae).
Separate assembly of the about 54 Mbp of sequence reads resulted in 7,759 unigenes for D. brevifolia , 12,546 unigenes for D. marnier-lapostollei and 19,143 unigenes

for F. rusbyi . Unigenes were screened for the presence of simple sequence repeats (SSRs, microsatellites) with motif lengths ranging from one to six base pairs.
Assuming a lower threshold of 15 repeat units for mono-, seven for di-, six for tri-, five for tetra-, and four units for penta- and hexanucleotide repeats, a total of 4,967 perfect SSRs were identified across the three resources.
The average frequencies were one SSR per 5.6 kb in both
D. marnier-lapostollei and F. rusbyi, and one SSR per 7.1 kb in D. brevifolia . Dinucleotide repeats were most abundant (54.3%), followed by trinucleotide (20.5%) and mononucleotide repeats (13.7%). Only perfect di- to hexanucleotide repeats were considered as candidates for
SSR marker development. Sufficient flanking sequence of appropriate quality and length for primer design proved to be present in 3,034 unigenes, harbouring a total of 3,743 SSR loci. Twenty perfect trinucleotide repeat loci were randomly selected from each of the three species, and tested for marker performance in the source species as well as in a wide range of bromeliads from various genera and subfamilies. The cross-species applicability of these markers for population genetics and their potential to differentiate between closely related species in highly speciose genera of the Bromeliaceae will be discussed.
P0431 – ePoster
Transcript profiling of CslF, CslH & Glb1 genes in developing barley endosperm
Wong, SC 1,2 , Burton, R 1 , Fincher, G 1 , Mather, D 2
1 ARC Centre of Excellence for Plant Cell Walls, School of Agriculture Food and Wine, Waite Campus, University
2 of Adelaide, Urrbrae, Australia; School of Agriculture
Food and Wine, Waite Campus, University of Adelaide,
Australia
Barley grain
β
-glucan concentration differs among varieties and affects end-use quality. The CslF and CslH cellulose-synthase like genes are involved in
β
-glucan synthesis (Burton et al. 2008; Doblin et al. 2009). The
Glb1 gene, which encodes a (1,3;1,4)-
β
-D-glucan endohydrolase is involved in the degradation of
β
-glucan and may also be involved in
β
-glucan synthesis.
Quantitative trait loci (QTLs) affecting grain
β
-glucan concentrations have been mapped with some QTLs colocating with Csl and Glb genes. The aim of this research was to investigate whether the parents of grain
β
-glucan mapping populations differ from each other (Steptoe vs
Morex, CDC-Bold vs TR251) with respect to CslF, CslH and Glb1 transcriptional profiles during endosperm development. As expected from previously published data (Burton et al. 2008), CslF6 had the highest transcript levels, but no marked differences in transcript levels were found between the parental lines. Therefore, current results showed that differences in transcript profiles of
CslF, CslH and Glb1 genes are not sufficient to explain
QTLs for grain
β
-glucan concentration in barley.
P0432 – ePoster
Comparative chloroplast genomes of Pinaceae: insights into the mechanism of diversified genomic organizations
Wu, C-S 1
M 1
, Lin, C-P 1 , Hsu, C-Y 1 , Wang, R-J 2 , Chaw, S-
1 Biodiversity Research Center, Academia Sinica, Taipei;
2 South China Botanical Garden, the Chinese Academy of
Sciences, China
Pinaceae, the largest family of the conifers, has diversified cpDNA organizations with two highly reduced large inverted repeats (IRs). To unravel the mechanism of this genomic diversification, the cpDNA organizations from 53 species of the ten Pinaceous genera were examined, of which, three species, viz.:
Larix decidua (122,474 bp), Picea morrisonicola
(124,168 bp), and Pseudotsuga wilsoniana (122,513 bp) were firstly elucidated. The results indicated that four distinct cpDNA forms (A-C and P) could be recognized by the rearrangements of two ~20 and ~21 kb specific fragments. The C form was documented for the first time and the A form might be the most ancestral one. Only the individuals of Pse. macrocarpa and Pse. wilsoniana were detected to have isomeric cpDNA forms. Three types
(types 1–3) of conserved repeats situated nearby the rearranged fragments were found to be syntenic. We hypothesize that type 1 (949 ± 343 bp) and type 3 (608 ±
73 bp) repeats are substrates for homologous recombination (HR), whereas type 2 repeats are likely inactive for HR because of their relatively short sizes
(151 ± 30 bp). Conversions among the four distinct forms may be achieved by HR and mediated by type 1 or 3 repeats, thus resulting in increased diversity of cpDNA organizations. We propose that in the Pinaceae cpDNAs, the reduced IRs might inactivate IR-mediated HR, then decreasing the diversity of cpDNA organizations, but the conserved repeats that the evolution endowed Pinaceae complement the reduced IRs and increase the diversity of cpDNA organizations.
P0434 – ePoster
1
Whole genome sequencing for crop improvement
Yang, B 1
BGI, Shenzhen, China
Obtaining the genome sequence of a wide range of individuals of a species will generate vast amounts of informative datasets and enable the rapid discovery of much greater genome-wide sequence variation than has been identified previously. With the decreasing cost of sequencing, the genetic maps of many species are getting increasingly dense, a great improvement for plant breeding and selection. Also, a wealth of knowledge will be gained from comparative genomic analyses within and across species, as how plants grow, function and survive different ecological conditions and various environmental stresses. To assemble large genomes from short-read sequences, BGI has developed a novel method for de novo assembly and has successfully assembled the cucumber (367Mb), Chinese cabbage (500Mb), and potato (830Mb) genomes. This method provides a cost effective approach to decode unexplored reference genomes and to carry out accurate analysis.
Resequencing individuals or populations with a reference genome can identify genetic variations and genes of important traits. Studies have been successfully
481

performed in maize, rice and soybean where genes related to growth, architecture, maturity, productivity or resistance were found. Most recently, we are promoting de novo sequencing of clade genomes, which will unveil the set of whole genome sequences in the same family or genus. Investigation of clade genomes will provide unique insight into mechanisms of the speciation process and functional elements that drive diversity in these genomes. Candidate genes related with important characteristics could be identified and further applied in plant molecular breeding. Whole genome sequencing will have far reaching implications for improving breeding strategies and plant varieties to meet the world's growing demand on plant production.
P0435 – ePoster
The stability of transgene expression and effect of
DNA methylation on PTGS in birch
Zeng, F 1 , Zhan, YZ 1
1 College of Life Science, Northeast Forestry University,
China
Transgenic plants of Betula platyphylla Suk. were obtained via Agrobacterium (LBA4404 strain)-mediated transformation. The transgene construct was pCAMBIA-
2301, which contained the selectable marker npt¢ò gene and reporter gene gus, along with a fused bgt gene consisting of an insecticidal toxin gene from a spider
( Atrax robustus ) and the C terminal of the Cry IA(b) gene from Bacillus thuringiensis. Transgenic plants were established by in vitro propagation and then, together with non-transformed control plants, were cultivated in a greenhouse under natural daylight conditions. We selected transgenic birch ( Betula platyphylla Suk) plants, which included non-silencing plants, the transcriptional silence plants included TP96, TP74, TP73 and the posttranscriptional silence ones (TP67 and TP72). The transcription of the bgt gene in different tissues and organs were significant difference. The transcriptional level of bgt gene in different tissues and organs was in the following order: leaf> female flower and male flower> branch bark> phloem > root. The transgenic lines were monitored for foreign gene expression for a long-term period of 8 years during their continuous growth under field conditions. GUS protein expression was not reactivated in transgene silencing lines TP72 and
TP67 when cultured in field conditions for long-term period. Meanwhile, no cases of gene silencing were observed again during the study period in field conditions. Our results suggest that transgene expression in transgenic birch plants appears to be stable under field conditions. The frequencies of methylated cytosines in the code regions of gus gene was studied. Relation of transgene expression and DNA methylation was analysed. The data of restriction enzyme digestion
(Hpa¢ò and Msp¢ñ) indicated that DNA methylation resulted in PTGS in transgenic birch.
P0437 – ePoster
Intergeneric crosses to obtain hybrids between
Salix
X
Populus
Zenkteler, M 1
Zadworna, A 1 ,
, Wojciechowicz, M K 1
Zenkteler, E 1
, Bagniewska-
1 Dept of General Botany, University of Poznañ, Poznañ,
Poland
The substitution of fossil fuels are the plant for biomass production, including woody species such as Salix and
Populus . The cross-linking between willows and poplars may provide the breeders attempts to put together the best characteristics of both genera (fast rate of growth and high biomass) into offspring usable as a renevable source of energy. Intergeneric crosses for gene recombination play a key role in plant biomass breeding program. Pollination, pollen germination and sigma receptivity of Salix (a forenoon 7.00 – 11.00 pattern of anthesis) are examined. Most of the pollen tubes failed to fertilize after exhibiting distorted growth patterns, resulted from incompatibility reactions. Only a few pollen tubes grew towards the ovules and were occasionally seen penetrating the embryo sac. Embryos at 21–28 DAP obtained after cross pollination between
Salix viminali s? X Populus tremula ? were isolated and rescued on ½ MS medium. Mature embryos were studied with light and fluorescent microscopy in comparison with the control crosses of Salix viminalis X Salix viminalis embryos. Intergeneric cross-embryos showed numerous post-fertilization phenomena begining from funiculus malformation and ending on the irregularity of cell dividing direction and frequency during embryo development. Results of the controlled pollination indicated that crossing was possible for the period between 1d before and 2d after anthesis Only 005 % of cross-pollinations were successful and low mature embryo set to be due to complex genetic factors, including incompatibility mechanism. Different morphologies were found in those intergeneric hybrids including stem high and leaf shape. To confirm the genetic background of intergeneric hybrids, RAPD reaction and Jaccard's coefficient of similarity was used to analyze the inheritance of hybrids (F1). For the RAPD hybrids the banding pattern of few hybrids had biparental inheritance and some hybrid had maternal inheritance.
Unequal crossover or gene conversion occurred during the intergeneric hybridization on the hybrid with maternal inheritance. In this study, based on the molecular data, intergeneric hybridization and embryo rescue was successful.
P0438 – Poster
Study of cellular localization of OsSUMOs in onion cells
1
Attia, K 1 , Saitoh, T
T 1 , Kimiko, I 1
1 , Igarashi, Y 1 , Kitajima, A 1 , Mitsui,
Niigata University, Japan
Small Ubiquitin-related Modifier (SUMO) is a member of an ubiquitin-like protein family that posttranslationally regulates cellular function of a variety of target proteins. SUMO conjugation mechanism to a protein substrate (Sumoylation) in plants and other organisms is analogous to that of the ubiquitin system, which requires the sequential function of the E1 (SUMO activation enzyme), the E2 (SUMO conjugation enzyme)
482

and the E3 (SUMO ligation enzyme). However, the biological consequence of SUMO modification is quite different from that of the ubiquitin that may Sumoylation against to ubiquitination and attenuates protein degradation. In plants, Sumoylation effector proteins are conserved. Recently, five putative SUMO genes have been found in the rice genome named OsSUMO1~5.
OsSUMOs have C-terminal diglycine (GG) motif as processing site, and three of them have SUMO consensus sequence ĵKXD/E. OsSUMO1 and OsSUMO2 lack the
SUMO consensus sequence and are homologous to human SUMO2/3 and Arabidopsis SUM1/2. OsSUMO2 protein has 90% identity to OsSUMO1 protein. To identify cellular localization of the OsSUMO proteins in plant, the expression vectors were constructed for constitutive RFP:OsSUMOs or DsRed:OsSUMOs expression and co-transfected with various construct expressing organellar marker:GFP fusion proteins into onion (Allium cepa) epidermal cells using particle bombardment technique. Laser scanning microscopic analyses of the cells revealed that DsRed:OsSUMO1 and
DsRed:OsSUMO2 are mainly localized in the nucleus and also in the cytoplasm. On the other hand,
DsRed:OsSUMO3 is only localized in nucleus and the
DsRed signal forms the dot-like subnuclear domain in the nucleus. Keywords: SUMO, RFP, GFP, onion cell, rice
P0439 – Poster
Revealing the yield impacts of organ-level quantitative trait loci associated with drought response in maize – a gene-to-phenotype modelling approach
3
1
Chenu, K 1 , Chapman, S
Welcker, C 3 , Hammer, G 1
QAAFI, Toowoomba, Australia;
INRA; 4 DEEDI
2 , Tardieu, F
2
3 , McLean, G 4 ,
CSIRO, Australia;
Substantial genotype-environment (GxE) interactions make crop improvement for drought tolerance a major challenge for plant breeding. As part of understanding and exploiting GxE interactions for yield, we propose a modelling approach to bridge the gene-to-phenotype gap and evaluate the influence of quantitative trait loci (QTL) for two key processes (leaf and silk elongation) that influence crop growth, water use and grain yield in maize. The yield of hypothetical recombinant inbred lines (RIL) was simulated for a range of climatic and drought conditions. QTL for faster leaf elongation increased crop yield under well-watered or pre-flowering deficit conditions, but reduced yield in terminal stress environments, as such genotypes prematurely exhausted their water supply. The QTL impact on yield was substantially enhanced when the potential pleiotropic effects on silk elongation were included. Simulations with this relatively simple model demonstrated the difficulty of interpreting the genetic control of yield at crop level. The results highlight the potential of integrative modelling for gene-to-phenotype prediction and for understanding GxE interactions for complex traits such as drought tolerance.
P0440 – Poster
Involvement of plant CLE peptide signaling in nematode infection process
Chika, E 1 , Ngan, BT 1 , Sawa, S 1
1 Kumamoto University, Japan
CLE peptide hormone is the molecule responsible for the control of plant meristem activity, and CLE genes are conserved in many plants. On the other hand, in mammals, only nematode that is infective to plants has
CLE gene. This CLE gene of nematode is likely to be functional in plants. Once nematodes infect roots, nematodes make root cells into the multinucleated giant cells, as source of nutrition, by injection of various redifferentiation factors. The CLE peptides are thought to be present in those redifferentiation factors. In this study, to elucidate the molecular mechanism of the infection process of nematode, nematode infection experiments in plants were performed. First, root-knot nematode
(Meloidogyne incognita), that inhabit in areas south of in
Japan and south east Asia, were infected with lots of wild-type strains and cultivars of Tomato. Compared with infection rate as an index of root-knot number, it was revealed that Micro TOM ( Solanum lycopersicum ),
Solanum pennellii, Solanum peruvianum showed resistance to the nematode infection. Following that, we identified and analyzed the sequences of Tomato homologue genes of CLV2, RPK2, SOL2, and BAM1, which are involved in CLE peptide signaling of
Arabidopsis .
P0441 – Poster
Molecular analysis of phylogenetic relationships among Iranian
Satureja
species
Hadian, J 1
1 Shahid Beheshti University, Iran
This study was undertaken in order to reveal genetic relationships and diversity of Satureja species using
AFLP and SAMPL markers. In AFLP analysis 999 polymorphic bands were amplified using 11 primer combinations. For SAMPL analysis 12 primer combinations were used and 981 bands were amplified among of them only one monomorphic band observed.
Maximum number of bands per species was belong to S. sahandica in both AFLP (488) and SAMPL (373) marker systems. In both marker systems, all accessions were grouped according to their species with high bootstrap values. In most cases, species relationships in clusters are in congruent with geographical distribution or chemical characters. Beside efficient species differentiation, AFLP and SAMPL revealed high level of genetic diversity in the genus Satureja.
P0442 – Poster
Arabidopsis
SOL1 functions in the CLE19 peptide maturation step
Hamasaki, R 1 , Tamaki, T 2 , Betsuyaku, S 2 , Sawa, S 1
483

1 Kumamoto University, Japan;
Japan
2 University of Tokyo,
Arabidopsis genome encodes 32 CLE genes, and the
CLE are supposed to function as 12 or 13 amino acid peptides. In the maturation step, CLE pre-protein would be digested by an specific peptidase. sol1 mutant was originally isolated as an suppressor mutant which mutation repress the effects of CLE19 over expression.
SOL1 is already reported to encodes peptidase. Here we will introduce the molecular genetic and biochemical analyses of the SOL1. sol1 mutant showed resistance to the over expression of the CLE19 gene. However, it was sensitive against to the CLV3 gene over expression. sol1 showed sensitivity to both of the synthetic CLE19 and
CLV3 peptides. These results indicated that the SOL1 function only in the CLE19 maturation step.
Recombinant SOL1 protein had an activity to digest a synthetic peptides. Together with the results with physiological experiment, we will introduce about the
SOL1 function in the CLE peptide maturation step.
P0443 – Poster
Functional analysis of liverwort MpCLE1 and
MpWOX
Honda, H 1 , Ishizaki, K 2 , Tabata, R
2 , Sawa, S 1
1 , Jedrusik, N 3 ,
1
Zachgo, S 3 , Kawauchi, T
Kumamoto University, Japan; 2 Kyoto University, Japan;
3 University of Osnabrueck, Germany
In plant morphogenesis, CLE peptide and its downstream factor WOX transcription factors have played an important function. The Arabidopsis CLV3-WUS system has been most analyzed pathway in CLE peptides.
However, molecular mechanisms are still poorly understood. One reason is gene redundancy of CLE and
WOX genes. Marchantia , a liverwort, has very low gene duplication, and Marchantia is considered as a good model plant for the analyses of signaling factors. Here we isolated MpCLE1 and MpWOX1. MpCLE1 OX plants showed epidermal-less phenotype. On the other hand,
MpCLE1 knock down plants showed excess cell division on the epidermal tissues. MpCLE1 expression was observed in the epidermal tissues by an in situ mRNA hybridization test. Together with the molecular genetic analyses of MpWOX1, we will introduce about the function of MpCLE1 and MpWOX1 in Marchantia .
P0444 – Poster
Sound stimulates plant gene response and plant pathogenic fungi in wavelength specific manner
Jeong, M-J 1 , Lee, Se-B
, Park, S-C
1 , Lee, S-K 1 , Suh, EJ 1 , Hwang,
1
E-W 1
National Institute of Agricultural Biotechnology, Rural
Development Administration (RDA), Suwon, Republic of
Korea
We investigated whether sound could alter gene expression in plants. Using a sound-treated subtractive library, a set of sound-responsive genes in plants was demonstrated through mRNA expression analyses. Of
484 them, the rbcS and ald genes, which are light responsive, up-regulated their expression with sound treatment in both light and in dark conditions. This suggested that sound could be used as a gene regulator instead of light.
When we analyzed ald gene expression using various single wavelengths, a significant increase in mRNA levels was found at 125 or 250 Hz but decreased at 50
Hz, indicating that the gene responded to sound in a wavelength-specific manner. To determine whether the ald promoter respond to sound, we generated transgenic rice plants harboring the chimeric gene consisting of a
1,506-bp promoter fragment of the ald gene fused to
Escherichia coli GUS reporter gene. Analyses of mRNA expressison level of three independent transgenic lines sound-treated with 50 or 250 Hz for 4 h showed that the
Gus gene expression in all three transgenic lines was up regulated by 250 Hz, but down regulated by 50 Hz.
These results correlated with sound responsive mRNA expression pattern observed for the ald gene in rice plants, indicating that the 1,506-bp ald promoter confers sound-responsiveness on a reporter gene in transgenic rice plants. We also tried to use specific sounds to control pathogens by examining the growth inhibition of fungi and bacteria. When a single specific sound frequency was applied to Botrytis cinerea growing on solid medium, mycelial growth was inhibited and spore formation was reduced. The organs were also deformed.
In addition, the mycelia treated with sound showed decreased spore formation, indicating that the effect continued into the next generation. When cucumber leaves that were infected with B. cinerea were treated with the specific sound frequency, the symptomatic area was decreased significantly as compared to the control group. This demonstrates that a specific sound frequency might be used, not only to increase plant growth, but also to rapidly control disease and counter severe environmental stress. We have demonstrated that in transgenic plants, specific frequencies of sound can be used to regulate the expression of genes fused to the ald promoter. (* Dr. Soo-Chul Park is Co-corresponding author)
P0446 – Poster
Isolation of the rpk2 enhancer mutants in
Arabidopsis
1
Kiyohara, S 1 , Sawa, S 1
Kumamoto University, Japan
Compare to the animals that form the many organs during embryonic development, plant continuously produces organs by the function of the shoot apical meristem (SAM). CLV3 function to regulate the SAM activity, and we recently identified the LRR-RLK,
RPK2, as a third receptor of the CLV3. As for the root, clv2 and rpk2 were shown to have resistance to synthetic
CLE peptides suggesting their function in the regulation of the RAM activity. In order to identify novel factors that function in the regulation mechanisms of RAM, we performed enhancer mutant screening by using rpk2 mutant by using the CLE peptide resistance as an index.
Here we will introduce our screening status.

P0447 – Poster
Analysis of the natural genetic variation of phosphite sensitivity in
Arabidopsis thaliana
Kollehn, DO 1 , Hardy, GSEJ 1 , O'Brien, PA 1 , Berkowitz,
O 2
1 School of Biological Sciences and Biotechnology,
Murdoch University, Murdoch, A ustralia; 2 School of
Plant Biology and Institute of Agriculture, University of
Western Australia, Crawley, Australia
Phosphate (H2PO4 ), as a source of phosphorous is one of the most critical macronutrients for plants. However, it is frequently not readily available to plants as it is often found in low concentrations and bound to other components of the soil. Therefore plants have developed strategies to adjust to phosphate starvation with a range of adaptations such as alterations of primary root length, number of lateral roots or secretion of organic acids.
Additionally, phosphate-depleted plants increase the expression of genes involved in phosphate acquisition, e.g. phosphate transporters and purple acid phosphatases.
Phosphite (H2PO3 ) is a more reduced form of phosphorous than phosphate. Although physiologically inert and not metabolised, phosphite is able to suppress the phosphate starvation response which results in an exacerbated phosphate depletion leading to a inhibition of plant growth. Similarly, phosphite can lead to toxicity at elevated levels, likely through interference with phosphate-dependent reactions or remobilisation. These observations indicate a recognition and uptake capacity of the plant for phosphite similar to that of phosphate. In addition, phosphite is used as a fungicide against plant pathogens, especially oomycetes such as Phytophthora species, which have devastating effects in agriculture and ecosystems (e.g. Phythophtora Dieback). Phytophthora species are a major threat for Australia’s biodiversity with more than 3000 species under threat in the Western
Australia alone. So far the only reliable measure to control these pathogens has proven to be phosphite for which the mode of action is yet to be elucidated. To gain a better understanding on the effects of phosphite on plant growth and defence we have started to investigate the natural genetic variation of phosphite-sensitivity in
Arabidopsis thaliana . Findings from this research will improve our knowledge of the mode of action of phosphite on plant defence responses, and might also have implications for the understanding of phosphate signalling or metabolism. Investigations into the genetic background of phosphite-dependent adaptation have been made by observing the phenotypic responses of 19 different Arabidopsis accessions. We will present first results obtained by screening for root phenotypes on media containing a range of phosphite / phosphate ratios.
P0448 – Poster
Does the 'red' gene in grapes affect the flavour of wine?
Koschella, A 1,2 , Boss, P 1 , Ford, C 2 , Walker, M 1
1 CSIRO Plant Industry, Wine Innovation West, Waite
Campus, Urrbrae, Australia; 2 University of Adelaide,
School of Agriculture, Food and Wine, Wine Innovation
Central, Waite Campus, Australia
Red and white wines, made from red and white grapes respectively, have very distinctive flavour and aroma profiles. While this could be partially attributed to the different methods used to produce these wines, we are testing the hypothesis that some of these distinguishing attributes are due to the presence of different flavour and aroma precursors in red and white grapes, or their differing flavonoid compositions. Flavonoids are a group of compounds including anthocyanins, flavanols, and proanthocyanins (condensed tannins) which have been linked to important wine attributes including mouthfeel and overall quality. The red pigment found in the skins of red grapes is due to the presence of anthocyanins. The transcription factors VvMYBA1 and VvMYBA2 have been identified as important regulators of anthocyanin synthesis and the two genes encoding these factors are mutated in white grapevine cultivars. The present research has used both naturally occurring and transgenic mutant grapevines in which VvMYBA1 and VvMYBA2 gene expression has been altered, to establish if there is a link between berry colour and wine flavour and aroma.
Using transgenic Chardonnay grapevines altered to produce anthocyanins, as well as Shiraz vines with partial (rose Shiraz) or complete silencing of the
VvMYBA1 gene (white Shiraz) together with controls, wines have been produced using microfermentation methods within the laboratory. Naturally occurring
Cabernet Sauvignon mutant grapes possessing rose and white coloured phenotypes were also fermented.
Headspace solid-phase microextraction (HS-SPME), along with gas chromatography and mass spectrometry
(GC/MS) was employed to analyse the volatile composition of each wine, and significant differences have been identified. RNA was also extracted from the transgenic berries and transcriptome analysis was performed using microarrays. The results of this research suggests that the MYBA transcription factors in grapevine may act to regulate other biosynthesis pathways, either directly or indirectly, resulting in the presence of different flavour and aroma precursors within berries of differing colour.
P0449 – Poster
Interrelationships between genetical, biochemical and ecological peculiarities of
Hypericum maculatum populations from the Baltic region
Kupcinskiene, E 1
Budiene, J 2
, Zybartaite, L
, Judzentiene, A 2
1 , Zukauskiene, J
, Paulauskas, A 1
1 ,
1 Dept of Biology, Vytautas Magnus University, Kaunas,
Lithuania; 2 Institute of Chemistry, Vilnius, Lithuania
The majority of worldwide studies on Hypericum genus refer to the species Hypericum perforatum , as a consequence of its application in the traditional medicine and modern phytotherapy. In Baltic countries 2 species of Hypericum are widely distributed – H. perforatum and
H. maculatum , although genetics and biochemistry of H. maculatum has got less attention when compared to H. perforatum . Present work aimed at evaluation of genetic diversity and composition of essential oils of Hypericum maculatum populations growing in some districts of
485

Baltic region. The plant material was collected from 12 districts at full-bloom stage in June–July. Flowers and leaves were used for essential oil analyses. Essential oils were taken by hydrodistillation. Analyses by GC/MS were performed using a chromatograph HP 5890 interfaced to an HP 5971 mass spectrometer and equipped with a capillary column DB-5. For DNA analyses leaves from the top of the plants were used.
Genomic DNA was isolated employing NucleoSpin Plant
II kit. Random Amplified Polymorphic DNA as the most popular method for plant DNA analysis was used. For
RAPD analysis 6 oligonucleotide primers of 10 nt length were used: OP-A1, OP-A9, OP-B19, OP-C7a, OP-A4,
OP-C11. Totally, 123 constituents were separated and identified in the samples of the essential oils extracted from the flowers and the leaves of Hypericum maculatum . The components made up to 85.8–99.1 % of the total content. The main classes were monoterpene hydrocarbons and oxygenated sesquiterpenes for the oils extracted from the flowers and oxygenated sesquiterpenes were prevailing among the oils extracted from the leaves of Hypericum maculatum. Investigated populations differed more according to the flower components of essential oils when compared to the leaves respective data. Prevailing components of the essential oils of the flowers were
α
-pinene, n-nonane, globulol and 2-methyl octane. Prevailing components of the essential oils of the leaves were globulol, germakrene
D, spathulenol and caryophyllene oxide. Among 6 primers used (OP-A1, OP-A9, OP-B19, OP-C7a, OP-A4,
OP-C11) all appeared to be informative for the studies of genetic diversity of populations of Hypericum maculatum. Each primer generated from 1 to 15 individual bands per population and provided a distinct and reproducible pattern of the amplified PCR fragments.
For all (12) populations of Hypericum maculatum the number of polymorphic loci was 106. Among populations Nei's gene diversity ranged between 0.097 and 0.141, Shannon's information index ranged between
0.141 and 0.215, the number of polymorphic loci ranged
26 and 49, percentage of polymorphic loci was 24.5-46.2.
Differences in geography of populations was not the main source of genetic and biochemical differences, habitat peculiarities, both climatic and edaphic also anthropogenic influence might be important sources of the variation.
P0450 – Poster
Regulation of the shoot apical meristem by SEUSS and SEUSS-LIKE 2 in
Arabidopsis
Lee, JE 1 , Golz, JF 1
1 University of Melbourne, Parkville, Australia
Plant growth is dependent on the continued production of new organs and tissues from the shoot apical meristem
(SAM), the activity of which is maintained by signals arising both from within the SAM and from other regions of the plant. We have recently shown that the
Arabidopsis transcriptional repressor LEUNIG (LUG), co-regulator SEUSS (SEU) and related SEU-LIKE
(SLK1-3) proteins, and YABBY DNA-binding proteins are part of a regulatory complex that promotes postembryonic SAM activity non-cell-autonomously (Stahle et al. 2009). This complex is also likely to be involved in
486 embryonic SAM formation, as seu slk2 double mutant seedlings lack a functional SAM. To better understand the function of the LUG-regulatory complex during embryogenesis, we examined the expression of key developmental regulators in seu slk2 embryos. This analysis showed that, while expression of the stem cellspecific CLAVATA3 gene is both delayed and reduced,
SHOOTMERISTEMLESS (STM) expression is completely absent, which is likely to account for the meristemless phenotype of seu slk2 mutants. We also found a strong reduction in expression of CUP-SHAPED
COTYLEDON 2 within the presumptive shoot apex, which is consistent with the loss of STM expression. In addition, PIN1 localisation and DR5-GFP reporter expression analysis indicates that auxin transport and signaling are significantly altered during the early stages of embryogenesis. We propose that these auxin distribution defects contribute to the cotyledon outgrowth and vasculature development defects apparent in seu slk2 mutants. Given the SEU is known to interact with ETTIN
(ETT), a member of the auxin response factor (ARF) family of transcription factors (Pfluger and Zambryski
2004), we next considered whether SEU-related proteins interact more broadly with ARFs. Using the yeast twohybrid assay, we show that SEU and SLKs are capable of interaction with ARF2 and ARF4, as well as ETT. In view of these results, we are currently testing whether embryos lacking ARF activity show similar phenotypes to those of seu slk2 double mutants. To determine whether SEU and SLK2 have non-cell-autonomous activity during embryogenesis, we have placed a functional YFP-SLK2 fusion construct under the control of lateral organ- and meristem-specific promoters in seu slk2 double mutants and are currently testing these lines for complementation of the SAM defect. We present preliminary data from these transgenic experiments and discuss their implications for a model describing the role of the LUG-regulatory complex during embryogenesis.
Pfluger, J, and Zambryski, P (2004). Stahle, MI et al.
(2009).
P0451 – Poster
Field transcriptome revealed critical developmental and physiological transitions in japonica
rice
Sato, Y 1 , Antonio, B
Kusaba, M 3
1 , Takehisa, H 1
, Nagamura, Y 1
, Namiki, N 2 ,
1 National Institute of Agrobiological Sciences, Tsukuba,
Japan;
Japan;
2 Mitsubishi Space Software Co. Ltd, Tsukuba,
3 Graduate School of Science, Hiroshima
University, Higashi-Hiroshima, Japan
Plant growth depends on synergistic interactions between internal and external signals, and yield potential of crops is a manifestation of how these complex factors interact, particularly at critical stages of development. As an initial step towards developing a systems-level understanding of the biological processes underlying the expression of overall agronomic potential in cereal crops, we performed continuous gene expression profiling of the japonica rice cultivar Nipponbare from transplanting to harvesting in order to establish a transcriptome profile encompassing the entire growth phase in the field. The uppermost fully-expanded leaf in the main stem, representing the 1st leaf from 13-76 days after

transplanting (DAT) and the flag leaf from 83 DAT until harvesting, were sampled at 12:00 PM every 7 days, covering 17 different growth stages with three replicates.
The continuous gene expression profile revealed that two major transcriptome changes occurred in the leaves from transplanting until harvesting. The first major change was observed before the panicle differentiation, accompanied by the expression of RFT1, a putative florigen gene in long day conditions, and the downregulation of the precursors of two microRNAs. This transcriptome change was also associated with physiological alterations including phosphate-homeostasis state as evident from the behavior of several key regulators such as miR399, suggesting that the first transcriptome change involves not only the initiation of panicle development but also various aspects of the physiological stage, which might be prerequisite for proper flowering and later developmental stages. The second major transcriptome change occurred just after flowering, accompanied by the upregulation of several NAC transcription factor genes associated with leaf senescence. To further examine the role of formation of a very strong sink, i.e., developing seeds, in the second transcriptome change, we also performed expression profiling of leaf on three independent sterile-mutant lines. The fertile and sterile lines basically showed similar expression profiles at the same sampling time, but the transcriptome change in the fertile line was more rapid and greatly enhanced as compared to the sterile lines, indicating that the second major transcriptome change is associated with leaf senescence, which autonomously starts independent of the development of sink, but is accelerated by the sink formation. In conclusion, a field transcriptome obtained by a wide range of gene expression profiling provided not only baseline information for functional characterization of genes but also revealed critical developmental and physiological transitions involved in the expression of growth potential under natural field conditions. With the accompanying gene expression profile database, RiceXPro
(http://ricexpro.dna.affrc.go.jp/), our resources on gene expression profiling may contribute to innovative crop improvements that have not yet been tried in classical or molecular breeding.
P0452 – Poster
A chloroplast localized 5' to 3' exoribonuclease,
RNase J, is a key nuclease responsible for RNA surveillance
Sharwood, R 1,2
Stern, D 2
, Halpart, M 3 , Luro, S 2 , Schuster, G 3 ,
1 Hawkesbury Institute For Environment, UWS,
Richmond, Australia; 2 Boyce Thompson Institute for
Plant Research, Cornell University, Ithaca, USA; 3 Dept of Biology, Technion, Israel Institute of Technology,
Haifa, Israel
Chloroplasts are semi-autonomous organelles, derived from prokaryotes that perform photosynthesis and other essential cellular functions. The regulation of chloroplast gene expression has levels of complexity not found in prokaryotes, particularly post-transcriptional processes including maturation of polycistronic transcripts, RNA editing and intron splicing. These events are mediated by nucleus-encoded endo- and exo-ribonucleases, along with RNA-binding proteins. One unique enzyme recently discovered is RNase J, which possesses both endoribonuclease and a processive 5'-3' exoribonuclease activity. An ortholog of RNase J exists within higher plants, encoded in the nucleus and transported into the chloroplast. At low concentration, the recombinant
Arabidopsis RNase J displayed only 5'-3' exoribonuclease activity and it is insensitive to the number of phosphates at the 5'end of the RNA. To determine the in vivo role of RNase J in chloroplast RNA metabolism, knockdown material was generated using
Virus-Induced Gene Silencing. Plant material with a twofold decrease in RNase J at the mRNA level was chlorotic indicating that chloroplast gene expression was impaired. We discovered that both discrete and heterodisperse antisense RNAs overaccumulated in silenced material, which suggested that readthrough transcription products are substrates for RNase J. Using an assay based on sensitivity to mung bean nuclease, a ssRNA-specific enzyme, we found a high proportion of asRNA duplexed with the complementary sense RNA.
According to polysome analysis, this duplexed RNA prevents translation of sense transcripts and prevents synthesis of crucial chloroplast proteins. Taken together, these results suggest that RNase J is a key surveillance enzyme preventing accumulation of deleterious asRNA.
P0453 – Poster
Isolation of the clv2 enhancer mutants in
Arabidopsis
Shimizu, N 1 , Tabata, R 1 , Yamada, M
Yamaguchi, K 3 , Shigenobu, S 3 , Sawa, S 1
2 , Hasebe, M 3 ,
3
1 Kumamoto University, Japan;
NIBB
2 Duke University, USA;
Compare to the animals that form the many organs during embryonic development, plants continuously produces organs by the function of the shoot apical meristem (SAM). CLV3 function to regulate the SAM activity. Three receptor complexes, CLV1, CLV2-
CRN/SOL2, and RPK2 function to perceive the CLV3 peptide hormone. In order to isolate novel factors that regulate CLV3 signaling pathway, we conducted the enhancer screening of the clv2 mutant, by using CLE peptide resistance in root as an index. We isolated 40< clv2 enhancer mutants. In order to identify the causal genes, we used next generation sequencer, and we already re-sequenced about 50 enhancer mutants. Here we will introduce our recent status about our clv2 enhancer screening experiment.
P0454 – Poster
Isolation and characterization of clen1 (clavata enhancer1 )
Tabata, R
S 3
1 , Yamada, M
, Fukada, H 4
2
, Hasebe, M 3
, Yamaguchi, K 3
, Sawa, S 4
, Shigenobu,
3
1 Kumamoto University, Japan;
NIBB;
2 Duke University, USA;
4 University of Tokyo, Japan
Shoot and root meristems are continuously located at growing tips and give rise to all kinds of plant tissues.
487

Therefore, to understand plant development, it is important to examine the mechanisms of meristem development. In Arabidopsis , it has been known that
CLAVATA3 (CLV3) peptide negatively regulate the size of the shoot meristem via receptors like CLV1, CLV2,
CRN, and RPK2. Moreover, Over-expression of CLV3 or CLV3/ESR-like (CLE) and treatments with CLE peptides reduce the size of root meristem and the length of root, suggesting that CLV-like signaling pathway also acts in the root. To identify the new factors regulating meristem development, we performed the clv2 enhancer screening as an index of CLE peptides-resistance phenotype of root. We identified causative mutation of clen1 (clavata enhancer1) among clv2 enhancer mutants by a combination of rough mapping and deep-sequencing analysis. Examined T-DNA insertion mutants of this gene also showed a same CLE peptides-resistance phenotype of root. This gene encodes a certain signal transduction molecule. Here, we will report our recent results on a role of this causal gene in CLV signaling pathway and progress of biochemical analysis of CLEN1 protein.
P0455 – Poster
High-resolution gene expression profiling in rice root
Takehisa, H 1
Yamauchi, T 3
, Igarashi, M 1
, Antonio, B
Nakazono, M 3, 4
1
, Abiko, T 3
, Motoyama, R 1
, Nagamura, Y 1
, Sato, Y 1 ,
, Inukai, Y 4 ,
1 National Institute of Agrobiological Sciences, Tsukuba,
Japan; 2 Graduate School of Biological Sciences, Nara
3 Graduate Institute of Science and Technology, Japan;
School of Agricultural and Life Science, Tokyo
4 University, Japan; Graduate School of Bioagricultural
Sciences, Nagoya University, Japan
Plant roots perform many essential adaptive functions including water and nutrient uptake, anchorage to the soil and the establishment of biotic interactions in the rhizosphere. Although the genetic regulations involved in root development have been widely studied in the model dicotyledon plant species, Arabidopsis thaliana , not much is known in monocotyledonous plant species such as rice, maize, wheat etc. Laser capture microdissection
(LCM) is an alternative technology that enables isolation of specific cell types from tissue sections using microscope. In combination with microarray analysis, this can be used as an efficient strategy to characterize the global gene expression of specific cells and tissues.
Here, we performed comprehensive gene expression profiling of root tissues from Oryza sativa L. ssp. japonica cultivar Nipponbare using LCM and microarray analysis in order to understand the genetic regulation involved in root development. Seeds were germinated on a nylon net floated on distilled water in a controlledenvironment growth chamber (60% humidity; 14h light period at 30°C; 10h dark period at 25°C). After 3 days of germination, the seedlings were transferred on a nutrient solution and allowed to grow for another 7 days. The crown roots were collected and divided into eight sections with different developmental stage from the root tip to the basal zone representing the root cap zone, cell division zone, cell elongation zone, three root hair zones, and two mature zones. Then using LCM, the three distinct tissues in the cell elongation/root hair zone and
488 mature zone corresponding epidermis/exodermis/sclernchyma, to cortex, the and endodermis/pericycle/stele, respectively were further separated. RNAs extracted from each sample were used for one-color (Cy3) labeling and hybridization with the rice 4 X 44K microarray RAP-DB (Agilent). Analysis of the microarray data derived from various tissues revealed differential gene expression patterns associated with root development from the root tip to the basal end.
Clustering analysis showed preferential expression of genes in different tissues and at different developmental stages. In the root cap, carbohydrate metabolism related genes which may be involved in mucigel and border cell production were detected. Many DNA replication and translation related genes associated with cell division and cell differentiation were identified in the cell division zone. In the cell elongation zone and root hair zone, transporter genes that may be involved in transport and uptake of water and nutrient were observed. Moreover, highly expressed genes were identified in the epidermis/exodermis/sclernchyma tissues at the root hair induction region in the cell elongation and root hair zone, and in the endodermis/pericycle/stele tissues at the lateral root initiation region. A comprehensive gene expression profiling of root tissues derived from LCM-microarray analysis will be useful in elucidating the gene regulatory networks involved in root development and morphogenesis in rice.
P0456 – Poster
Comparative transcriptome analysis during seed development of viviparous rice
Huh, SM 1
DY 1
, Hwang, YS
, Yoon, IS 1
2 , Hwang, HJ 1 , Shin, YS 3 , Kim,
1 Natural Academy of Agricultural Science; 2 Dept of
Bioscience and Biotechnology, Konkuk University, Seoul,
Republic of Korea; 3 Rice Research Division, National
Institute of Crop Science, Suwon, RDA, Republic of
Korea
Preharvest sprouting (PHS, vivipary) is an important agronomic trait related to seed dormancy that results in significant economic loss. To identify genes involved in this process, comparative transcriptome analysis was applied to rice seeds from Korean cultivars presenting contrasting preharvest sprouting behavior. The results showed that 336 genes were up-regulated and 608 genes down-regulated in the viviparous seeds at 25 days and 40 days after flowering. These were C/N metabolic enzymes, storage proteins, transporters, stress-related proteins, protein kinases and transcription factors.
Several genes within the QTL region for seed dormancy were also included. Expression of key genes of ABA metabolism or signaling was not significantly different, and ABA inhibited the viviparous seed germination, indicating that the preharvest sprouting phenotype is not likely to be related to ABA. It is noted that genes for seed storage proteins and starch metabolism were more rapidly changed in the high viviparous rice at seed maturation stage. Our current work provides important information for cloning of seed dormant genes in rice.
Supported by a grant (PJ006684) and Postdoctoral
Fellowship Program of NAAS, RDA.

P0457 – ePoster
Molecular cloning of a stress-responsive aldehyde dehydrogenase gene ScALDH21 from the desiccationtolerant moss
Syntrichia caninervis
and its responses to different stresses
Yang, H 1 , Zhang, D 1 , Wood, AJ 2 , Zhang, Y 1
1 Key Laboratory of Biogeography and Bioresource In
Arid Land, Xinjiang Institute of Ecology and Geography,
Chinese Academy of Sciences, Xinjiang Urumqi, China;
2 Dept of Plant Biology, Southern Illinois University-
Carbondale, Carbondale, USA
Aldehyde dehydrogenases (ALDHs) are key enzymes of abiotic stress-tolerance in a variety of organisms. The
ALDH gene superfamily in eukaryotes has identified 12 protein families based upon sequence identity, among which, ALDH21 is unique to moss and represented by just one gene from the moss Tortula ruralis . In order to verify whether ALDH21 exists and has the stressresistant character, the techniques of homology cloning and anchored PCR were employed to clone the ALDH21 gene of Syntrichia caninervis (ScALDH21), an extremely desiccation-tolerant moss that survives in the
Central Asia desert. ScALDH21 gene was subcloned into pET26b (+) and expressed in E. coli (Rosetta) to test its function in response to desiccation and salinity.
Quantitative RT-PCR was used to analyze expression in response to abscisic acid (ABA) and desiccation. The
ScALDH21 cDNA was cloned and found to be 1,452 bp in length, which encodes a protein of 483 amino acids, approximately 97% identical to Tortula ruralis ALDH21
(TrALDH21A). The molecular weight of ScALDH21 protein is 53 kDa. ScALDH21 transcript levels were increased significantly in response to desiccation and
ABA. ScALDH21 protein could be induced under the salinity and desiccation stress in the transgenic E. coli , and was more abundant within salt-treated gametophytes, but not in the control strains. The data suggest that
ScALDH21 participates in the stress-resistant pathways and plays an important role in response to desiccation and salinity stresses.
P0458 – ePoster
Investigation of abiotic stress on die-back
Pistacia vera
L.
Abbaspour, H 1 , Saeidi-Sar, S 1
1 Dept of Biology, Damghan Branch, Islamic Azad
University, Damghan, Iran
Environmental stresses represent the most limiting factors for agricultural productivity. Apart from biotic stress caused by plant pathogens, there are a number of abiotic stresses such as drought and salinity which have detrimental effects on plant growth and yield. Pistachio is one of the most important horticultural products of Iran.
It has been embraced as one of the most commercial products. In the recent years, dieback has been a wide spread problem on pistachio planting gardens and has caused product reduction. In order to investigate the effect of salinity in dying those branches and define the distribution and the probability of occurrence of end searing, some experiments were done. In this study to define distribution and the probability of dieback, 130 selected pistachio orchards in Rafsanjan city were investigated. In addition, in order to find out the probability of occurrence and some affecting factors in dieback, 45 orchards were chosen and sampling of water, leaves and three different depths of soil was done. These results showed that the amount of pollution was varying between 17 to 96 percent and average of 29 percent was estimated. There was also a significant relationship between percentage of dieback, the amount of soil pH and relative percentage of clay. Also, correlation between other characteristics of soil and the results of water and leaves analysis were significant.
P0459 – ePoster
Effect of salt stress on lipid peroxidation, antioxidative enzymes, and proline accumulation in pistachio plants
Abbaspour, H 1 , Saeidi-Sar, S 1
1 Dept of Biology, Damghan Branch, Islamic Azad
University, Damghan, Iran
Abiotic stress, such as salt stress is serious threats to agriculture and the natural status of the environment.
Plants exhibit a variety of responses to abiotic stresses that enable them to tolerate and survive adverse conditions. The changes in lipid peroxidation, antioxidant enzymes, such as catalase (CAT), glutathione reductase (GR), superoxide dismutase (SOD), ascorbate peroxidase (APX), and proline accumulation in relation to the tolerance to salt stress in Pistacia vera L. was investigated. The 60 days old pistachio seedlings were subjected to four salt treatments (0, 100, 300, and 500 mM NaCl) for 23 days. Pistachio plant exhibited a decrease in lipid peroxidation in 300 and 500 mM NaCl.
Activities of CAT, SOD, and APX increased with increasing salt stress, but activity of GR only a slight decrease in 100 and 300 mM salt treatment and an increase under 500 mM salt treatment. The contents of proline in the leaves increased at all levels of salinity except at 100 mM. All the results show that in pistachio plants alleviates the deleterious effect of salt stress, through protection mechanism against oxidative damage by maintaining a inherited and induced activity of antioxidant enzymes and higher proline content that may play a role as an enzyme-stabilizing agent in salt stress.
P0460 – ePoster
Evaluation of salt tolerance in pistachio plants cv.
Akbari, Fandoghi and Kalaquchi by physiological characters under greenhouse conditions
Abbaspour, H 1 , Saeidi-Sar, S 1 , Afshar, H 2
489

1 Dept of Biology, Damghan Branch, Islamic Azad
University, Damghan, Iran; 2 Dept of Biology, Neyshabur
Branch, Islamic Azad University,. Neyshabur, Iran.
Salinity is known to adversely affect production of most agricultural plants, because their biomass and seed yield are significantly suppressed by this abiotic menace. In this study, salt tolerance in pistachio plants cv. Akbari,
Fandoghi and Kalaquchi were evaluated. Salt induced changes in growth, plant pigments, solute and ion accumulation in greenhouse experiment on three pistachio ( Pistacia vera L.) cultivars. Pistachio plants were grown under NaCl (0, 100, 300 and 500 mM) for two month. Salt stress markedly reduced growth, leaf area and content of pigments in cultivars of Fandoghi and
Kalaquchi but not significant effects on cv. Akbari. The negative effects of medium and high salinity were not significant on free proline, K + , P and Cl content of cv.
Akbari but that was significant on cv. Fandoghi and
Kalaquchi. Also, low salinity had negative effects on cv.
Kalaquchi but not on other cultivars. Based on these results, cv. Akbari was found to be salt tolerance, cv. fandoghi moderately tolerant and cv. Kalaquchi salt sensitive.
P0462 – ePoster
Effect of drought stress on the antioxidant activity, lipid peroxidation, some physiological parameters, and composition of essential oil from
Mentha pulegium
L.
Ahmadiani, S 1 , Fshar Mohammadian, M 1 , Ghanati, F 2
1 Dept of Biology, Faculty of Science, Guilan University,
2 Rasht, Iran; Dept of Plant Biology, Faculty of
Biological Science, Tarbiat Modares University, Tehran,
Iran
Lamiaceae is a large family composed of different genus with valuable medicinal properties in Iran. Mentha pulegium belongs to lamiaceae and is important for its antibacterial and antioxidant characteristics. It can be found in different areas in the north of Iran. Ecological stresses such as drought, salt, and climate conditions may affect physiological activity of the plants. In the present study, the seeds of Mentha pulegium were collected from
Rasht and Lahijan. The seedlings were grown hydroponically and then were treated with PEG 5% in order to induce drought stress. Essential oil were prepared from the aerial parts including leaves of this plant and Clevenger-type apparatus was used to prepare essential oil from dried sample in double distilled water.
The effects of drought stress on the activity of catalase, peroxidase, superoxide dismutase, ascorbate peroxidise, membrane lipid peroxidation as well as other physiological parameters and some composition of essential oil were subjected to study. The results indicated that drought stress increased the activities of catalase and peroxidase in roots of both plants originated from Rasht and Lahijan, significantly. Drought stress increased the activities of ascorbate peroxidase in roots of plants originated from Lahijan. Significant increase observed in MDA of PEG-treated plants originated
Rasht. Additionally, increase of glucose, galactose, rhamnose, and xylose, chlorophyll b and carotenoid content were significant in PEG-treated plants from both areas.
P0465 – ePoster
Growth-related gene expression in desiccated resurrection plants
Blomstedt, C 1 , Griffiths, C 1 , Islam, S 1 , Hamill, J
1
D 1 , Neale, A 1
Monash University, Australia
1 , Gaff,
Molecular studies have been conducted in the resurrection plant S. stapfianus to investigate the molecular and cellular mechanisms utilised by these desiccation tolerant plants to cope with severe waterdeficit. S. stapfianus is a versatile research tool for investigating desiccation-tolerance in vegetative grass tissue, with several useful characteristics for differentiating desiccation-tolerance adaptive genes from the many dehydration-responsive genes present in plants.
A number of genes orthologous to those isolated from dehydrating S. stapfianus have been successfully used to enhance drought and salt tolerance in model plants as well as important crop species. In addition to the ability to desiccate and rehydrate successfully, the survival of resurrection plants in regions experiencing short sporadic rainfall events may depend substantially on the ability to tightly down-regulate cell division and cell wall loosening activities with decreasing water availability and then grow rapidly after rainfall while water is plentiful. An analysis of the growth capacity of S. stapfianus following a dehydration-rehydration treatment indicates regrowth is substantially more rapid in rehydrated plants compared to well watered plants. Our analysis of gene expression in S. stapfianus has revealed that transcripts from several important growth-related genes are present in the desiccated tissue of this resurrection plant at elevated levels, which supports the proposition that resurrection plants may be a useful tool for pinpointing genes associated with regulation of growth rate and biomass production.
P0466 – ePoster
Investigating a genetic basis to physiological responses of
Eucalyptus globulus
seedlings to defoliation
1
Borzak, CL 1,2 , O’Reilly-Wapstra, JM
Pinkard, EA 4 , Potts, BM 1
1 , Barry, KM 3 ,
School of Plant Science, University of Tasmania,
2 Hobart, Australia; CRC for Forestry, University of
Tasmania, Hobart, Australia; 3 Tasmanian Institute of
Agricultural Research, University of Tasmania,
Australia; 4 CSIRO Ecosystem Sciences, Hobart,
Australia
Eucalypt seedlings have a remarkable ability to recover after mammalian browsing damage. Plant recovery mechanisms may include physiological responses such as increased growth rate, changes in resource allocation patterns and changes in photosynthetic rates. Previous studies have shown that Eucalyptus globulus seedlings can compensate for herbivory through increased
490

photosynthetic capacity, however, the degree of genetic control of this physiological mechanism is unknown. In this study we investigated the genetic basis to variation in growth and physiological responses of E.
globulus seedlings to artificial defoliation using seedlings sourced from eight families, from three populations representing relatively high, intermediate and low chemical resistance to mammalian herbivory. Photosynthetic rates of seedlings were measured before and after a defoliation
(50% of leaves artificially removed) treatment across different time periods. Results shed light on the genetic basis to recovery mechanisms of E. globulus after loss of foliage, as well as trade-offs between plant recovery and chemical resistance.
P0467 – ePoster
1
Acquisition of desiccation tolerance and dormancy in seeds of
Erythrina speciosa
, a native tree of the
Brazilian Atlantic Forest
Braga, M 1 , Kretzschmar, F 2 , Centeno, D 3
Bioquímica, Sao Paulo, Brazil;
, Barbedo, C 4
Instituto de Botanica, Nucleo de Fisiologia e
2 SABESP, Brazil;
3 Universidade Federal do ABC, São Bernando do
Campo, Brazil; 4 Instituto de Botanica, Nucleo de
Sementes e Melhoramento Vegetal, São Paulo, Brazil
Orthodox seeds have varying degrees of sensitivity to water loss during their maturation and therefore are interesting models for studying desiccation tolerance.
Erythrina speciosa Andrews (Fabaceae) is a species native of Atlantic forest that, although is found in flooded areas of southern and southeastern Brazil, produces orthodox seeds with high tolerance to desiccation. Relationships between desiccation tolerance and dry matter, water, and sugar contents were studied throughout the seed development of E. speciosa
Andrews. Seeds of E. speciosa were separated into 6 stages of maturation by size and pigmentation and hardness of the seed coat and analyzed for germination capacity, water content, dry matter, water potential, and soluble sugar content and composition. The water potential and water content of seeds decreased gradually from the first stage to the last, accompanied by increase in dry weight. The physiological maturity was reached at stage IV, coinciding with the highest accumulation of dry mass. The seeds were only able to germinate from the third stage (12%), reaching 100% only in the last stage, when dormancy is observed due to tegument impermeability. Reduction in water content down to
14%, observed in the stage III, was enough to promote seed germination. The seeds were also subjected to osmoconditioning with polyethylene glycol (PEG), and results indicate that in the third stage of maturation E. speciosa seeds are sensitive to water deficit, increasing its germination capacity, but still do not tolerate drying at very low water content. Sugar analyses revealed high proportion of cyclitols throughout development, reaching
85% of total sugars tested, and decreased gradually during maturation (~ 50%). Glucose and fructose remained low in both axis and cotyledons during the first five stages of development and increased slightly in the mature seeds. Raffinose and stachyose gradually increased in axes and cotyledons from stage III and IV, respectively, until the sixth stage, with greater increase in the fourth stage when seeds are tolerant to drying at 10% water. These results suggest that raffinose and stachyose are implicated in the acquisition of tolerance desiccation and the acquisition of germination capacity in seeds of E. speciosa , a species that occurs in an environment with high water availability. (FAPESP, CNPq)
P0470 – ePoster
Vascular functioning and the water balance of ripening kiwifruit (
Actinidia chinensis
) berries
1
Clearwater, M 1
Thorp, G 2
, Luo, Z 2 , Chye, S 3 , Blattmann, P 3 ,
University of Waikato, Hamilton, New Zealand; and Food Research, Auckland, New Zealand;
Food Research, Te Puke, New Zealand
3
2 Plant
Plant and
Indirect evidence suggests that water supply to fleshy fruit during their final stages of development occurs through the phloem, with the xylem providing little water, or acting as a pathway for water loss back to the plant. This inference was tested by examining the water balance and vascular functioning of ripening kiwifruit berries ( Actinidia chinensis var. chinensis ‘Hort16A’) exhibiting a late season ‘shrivel’ disorder in California, and normal development in New Zealand. Dye labelling and mass-balance experiments indicated that the xylem and phloem were both functional and contributed approximately equally to the fruit water supply. The modelled fruit water balance was dominated by transpiration, with net water loss under high vapour pressure deficit (Da) conditions in California, but a more positive balance under cooler New Zealand conditions.
Xylem hydraulic conductivity measured using a passive evaporative method was an order of magnitude lower than an estimate obtained from the pressure infusion method used previously with other fruits, but yielded more realistic predictions of xylem flows. Direct measurement of pedicel sap flow under controlled conditions confirmed inward flows in both the phloem and xylem. Phloem flows were required for growth, with gradual recovery after a step increase in Da. Xylem flows alone were unable to support growth, but did supply transpiration and were responsive to Da induced pressure fluctuations. Results suggest that the shrivel disorder was a consequence of a high fruit transpiration rate, and that the perception of complete loss or reversal of inward xylem flows in ripening fruit should be re-examined.
P0471 – ePoster
The challenge of characterizing and quantifying the different cell wall polysaccharides in a range of plant species and tissues
Collins, H 1,2
Fincher, G 2
, Lahnstein, J 2 , Burton, R 2 , Fabrizio, J 1 ,
1 ARC Centre of Excellence In Plant Cell Walls,
University of Adelaide, Australia; 2 CSIRO Food Futures
Flagship, High Fibre Cluster, Australia
Cell walls are an important component of all plants, conferring strength to support the plant and flexibility for the plant to grow. Cell wall polysaccharides are the main
491

component of dietary fibre. They are resistant to digestion in the human small intestine and are completely or partially fermented in the large intestine. They are also an abundant source of renewable biopolymers and have the potential to be used for biofuel production. Therefore, a greater understanding of the abundance and structure of these polysaccharides in plants, in-particular cereals, is an important research aim. The main non-cellulosic cell wall polysaccharides are xylans, (1,3;1,4)-
β
-D-glucans, mannans and xyloglucan. Of particular interest to us are the most abundant cell wall components in the endosperm of cereals, the arbinoxylans and (1,3;1,4)beta-D-glucans. The amount of (1,3;1,4)-
β
-D-glucans found in cultivated cereals ranges from less than 0.02% in rice to 3-7% in oats, with some barley cultivars containing as much as 20% (Collins et al. 2010), while the endosperm of the model cereal Brachypodium contains about 40% (1,3;1,4)-
β
-D-glucans (Guillon et al.,
2010). Arabinoxylans can vary from 1.8% in sorghum grain to 7-12% in rye (Collins et al. 2010). This range in the amounts of these polysaccharides has led to the development of analytical methods for the comparison of amounts in different plant species. In addition, plant tissues can contain large amounts of compounds that interfere with the analysis of the target polysaccharide.
For example, there are large amounts of starch in the endosperm of rice compared to the very low levels of
(1,3;1,4)-
β
-D-glucans. This has led to the need for clean up procedures to remove starch and other products.
These methods are robust and sensitive and are used routinely in our laboratory.
P0475 – ePoster
Development of a new model system for analysis of cell wall biosynthesis genes
Dimitroff, G 1 , Little, A 2 , Burton, RA 1 , Fincher, GB 1
1 ARC Centre of Excellence in Plant Cell Walls,
Australia; 2 School of Agriculture, Food and Wine,
University of Adelaide, Australia
Plant cell walls are diverse, complex structures consisting of many different cellulosic and non-cellulosic polysaccharides. The cellulose synthase-like CslF genes have been shown to be required for the biosynthesis of
(1,3:1,4)-
β
-D-glucans (Burton et al. 2006), which are major cell wall constituents in cereals and grasses.
Investigations into the specific end products of each gene have involved QTL mapping, random mutagenesis and targeted creation of transgenic overexpression/silencing lines. However, in order to understand the roles of each gene in more detail it is important to be able to express them individually and in different combinations in a transient system devoid of endogenous (1,3:1,4)-
β
-Dglucans. Nicotiana benthamiana proved to be an ideal host due to the lack of detectable (1,3:1,4)-
β
-D-glucans, ease of transient transformation and presence of substrates required for cell wall polysaccharide synthesis.
A transient expression system was setup using
Agrobacterium injection into the leaves and has proved quite successful in obtaining some preliminary results.
However, as monocot-specific genes are being tested in a dicot system, there are a number of technical issues such as the availability of substrate, possible ancillary proteins and post-translational modifications that are decreasing
492 the reliability of the results. Therefore, a more closely related model system is required to eliminate these issues. Studies of rice have shown that the endosperm contains extremely low levels of
β
-glucan. Therefore as this is a cereal, a callus or suspension culture line derived from this cell type should contain all the factors necessary with minimal interference from background polysaccharide levels to obtain more informative results with respect to the roles of the individual CslF genes.
P0479 – Poster
Ethylene and nitric oxide effect in
Catasetum fimbriatum and
Dendrobium
Second Love
(Orchidaceae) etiolation
Félix, L 1 , Chaer, L 1 , Kerbauy, G 1
1 University of São Paulo, Brazil
Catasetum fimbriatum a Brazilian epiphytic orchid, has been used for many years at University of São Paulo plant physiology laboratory as plant model for physiological studies of plant development due its unusual meristematic characteristic. This plant when aseptically incubated in dark presents an etiolated shoot phenotype, which can be used to clone the plant from its lateral bud segments, drastically reducing somaclonal variations and promoting genetic stability. That is only possible because C. fimbriatum can successfully retake apical meristematic activity whenever in absence of light for undetermined time. Dendrobium Second Love is another epiphytic orchid that has important commercial and ornamental value, but contrarily to C. fimbriatum , this genera doesn’t maintain its apical meristematic activity for such long period and has limited shoot elongation when kept in dark. Several studies sustain that ethylene is involved in shoot elongation inhibition and there are experimental evidences that the signaling molecule nitric oxide participates in many chemical reactions and could be involved on shoot apical meristematic activity. Ethylene and nitric oxide could be acting directly or indirectly by affecting other hormones pathways such gibberellins. The purpose of this study is to verify the effect of ethylene and nitric oxide in both
Catasetum fimbriatum and Dendrobium Second Love after a month in light absence treated plants. A genetically uniform population of C. fimbriatum and
Dendrobium maintained in our laboratory was incubated in USP medium containing Fe-EDTA, supplemented with micronutrients of Murashige and Skoog (1962),
1mg/L thiamin, 1g/L soy peptone, 100mg/L inositol and
20g/L sucrose. The pH of the medium was adjusted to
5,8 before adding 2g/L Phytagel, and the medium was autoclaved for 15 min at 120° C. The flasks were sealed and cultures were then submitted to dark in a constant temperature of 26° C ± 1° C for 30 days. 48 hours before ethylene and nitric oxide analyses flask internal air was renewed with synthetic air. A flame ionization detector was employed for ethylene analyses and nitric oxide production by the plant shoot tissues was determined spectrofluorimetrically by using 4,5-diaminoflurescein as a specific nitric oxide probe. When compared to plants of
C. fimbriatum , Dendrobium plants presented a reduced shoot length and lateral buds liberation. Such characteristics are very similar to those observed in plants treated with ethylene in sealed flasks.

Corroborating to this observed phenotype, ethylene emission was significantly higher in plants of
Dendrobium than in C. fimbriatum . However,
Dendrobium shoot tissue presented higher levels of nitric oxide than C. fimbriatum , this result may be a sign of nitric oxide effect in shoot elongation inhibition in
Dendrobium plants. As gibberellins are known to be one of the most important hormone in shoot elongation in plants submitted to light, we will aim our researches to verify what is the real relation between those compounds and gibberellins regarding plant etiolation.
P0480 – Poster
The relationship between ethylene, nitric oxide, carbon dioxide emission and etiolation of
Catasetum
1 fimbriatum
,
Paphiopedilum and
Cymbidium
(Orchidaceae)
Chaer, L 1 , Felix, LM 1 , Kerbauy, GB
University of São Paulo, Brazil
1
Catasetum fimbriatum (Morren) Lindl. is an epiphytic orchid that has been studied since 1990 at our
Laboratory. This plant has the peculiarity of being easily propagated, using the lateral buds of its etiolated shoot.
This is possible because the shoot apical meristem shows a sustained activity when maintained in darkness, resulting in an expressive shoot elongation. When the etiolated nodal segments are incubated under light, they rapidly form new plants. This micropropagation process dismisses the application of any plant growth regulators, representing an efficient in vitro multiplication method for Catasetum . Other more valuable genera on the ornamental plant market, such as Paphiopedilum and
Cymbidium , have shorter shoot elongation when cultivated in the dark, taking a long time to produce few nodes and, therefore, this is not an appropriate large scale micropropagation method. Previous studies with C. fimbriatum carried out at our laboratory have shown an inhibitory effect of shoot growth at sealed flasks without gas exchange in the absence of light, which is a similar effect of ethylene injection in this plants, indicating the involvement of this hormone in shoot apical meristem activity regulation. Other gas hormones or substances that are accumulated in sealed flasks might be involved in the regulation of shoot growth, such as the signaling molecule nitric oxide (NO) or carbon dioxide (CO
2
). The aim of this study is to understand the effects of ethylene,
NO and CO
2
emission in the etiolation process of
Catasetum fimbriatum , Paphiopedilum and Cymbidium maintained for 1 year in the absence of light. Plants were obtained by micropropagation technique, using etiolated nodal segments and grown in vitro for 3 months in 25ºC and 12 hours photoperiod. These plants were incubated in the dark for one year and flask internal air was renewed with synthetic air 48 hours before the analyses.
Ethylene and CO
2
samples of 10mL and 1mL respectively, were taken and gas composition was determined with gas chromatography. NO production by the plant shoot tissues was determinate spectrofluorimetrically by using 4,5-diaminoflurescein as a specific NO probe. The present study demonstrated that
Catasetum fimbriatum showed the highest shoot elongation while Paphiopedilum plants had the lowest.
On the other hand, Paphiopedilum plants had substantial levels of NO in tissue, which was higher than
Cymbidium , whilst Catasetum fimbriatum had the lowest
NO detection. Therefore, these results indicate that NO could, directly or indirectly, inhibit the shoot elongation of dark growing plants. Ethylene production was undetectable in Catasetum plants, as well was observed in Cymbidium and Paphiopedilum this may have occurred probably because the emission rate of this gas was low. CO quantification analyses showed no
2 significative differences between Paphiopedilum and
Catasetum plants, which showed a dramatically different shoot elongation, while Cymbidium had a higher CO
2 emission, indicating an elevated respiration rate. Thus, it is not possible to establish a relationship between CO
2 production and shoot elongation in these genera.
Nevertheless, the effects of different gases in meristematic activity of these plants cultivated in the dark will need more experiments for further conclusions.
P0481 – ePoster
Violaxanthin cycle can be activated in darkness by stresses such as desiccation and anoxia
1
Fernández-Marín, B
García-Plazaola, JI 1
1 , Míguez, F 1 , Becerril, JM 1 ,
University of Basque Country (UPV/EHU)
Violaxanthin (V) cycle is usually activated by light in plants and algae. Here other stressors that may trigger V de-epoxidation in darkness are characterized in the brown intertidal alga Pelvetia canaliculata . Thalli collected in the field were exposed to high light, or to different stressing factors in darkness: desiccation, immersion and anoxia. Violaxanthin de-epoxidation and maximal photochemical efficiency of PS II (Fv/Fm) were monitored during these experiments. All the stressors induced a de-epoxidation of V-cycle in the same extent, being fully reversible after the cessation of the treatments. Fv/Fm followed the reverse pattern reaching the minimum values when maximum de-epoxidation degree was observed. Taken together all results, and irrespective of the treatments applied, a negative linear relationship was obtained between the de-epoxidation of
V and Fv/Fm. These results indicate that stressors other than light (in particular anoxia and desiccation) are able to induce V-cycle de-epoxidation in P. canaliculata. A common mechanism different to light-induced proton pumping (presumably chlororespiration), is necessary to generate the acidification required for the activation of violaxanthin deepoxidase. Zeaxanthin formed under these different treatments seems to play the same role in the regulation of light energy conversion.
P0482 – ePoster
Analysis of storage carbohydrates in different organs of herbaceous species from rocky fields
Joaquim, EO 1 , Carvalho, MAM 1 , Figueiredo-Ribeiro,
1
RDC 1
Institute of Botany, Plant Physiol & Biochem Dept, São
Paulo, Brazil
493

In many plant species some organs perform more than one function at certain stages of the life cycle. Roots, stems or leaves begin to accumulate reserve substances and depending on the origin may be transformed into storage organs like tubers, corms, pseudobulbs, bulbs, rhizophores, rhizomes and tuberous roots. Geophytes, for instance, have evolved to survive adverse environmental conditions through the production of those underground storage organs, widespread in rocky fields. Among other storage compounds, carbohydrates assigned several functions such as source of energy and protection against drought, high salinity and extreme temperatures. The rocky fields on Brazil are characterized by mesothermal climate, with three to four months of a dry season referring to winter, and seven to eight months of humid season, referring to summer. The temperature ranges between 17 to 20°C and the annual rainfall is about 1500 mm. The average altitude is higher than 800 meters, the soils are typically shallow, sandy and with rocky outcrops. Generally, the flora of the rocky fields has a high degree of endemism. The aim of this work was to carry out a screening of reserve compounds accumulated in different organs of herbaceous species predominant in those fields. The plants were collected both at the 'Serra do Cipó' (MG- Brazil) and 'Serra Dourada' (GO-Brazil) regions. The percentage of soluble carbohydrates was determined by colorimetric methods and starch by enzymatic assays. Samples were also analyzed by high performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD).
Oncidium hidrophyllum (Orchidaceae) concentrates more than 24% of total soluble carbohydrates in the bulbs and another Orchidaceae, Habenaria caldensis, contains 41% of soluble carbohydrates in the tuberous roots. Both species presented glucose, fructose and sucrose among the main sugars. Roots of Actinoseris angustifolia , rhizophores of Vernonia linearifolia and Vernonia psilophyla , tuberous roots of Viguiera kunthiana and
Chresta speciosa, all Asteraceae species, accumulate fructose polymers, fructans of the inulin type, constituting 22%, 14%, 34%, 29% and 23% of total soluble carbohydrates, respectively. Fleshy leaves of
Leiothrix curvifolia (Eryocaulaceae) presented low amounts of soluble sugars, predominantly glucose.
Despite of the importance of carbohydrate metabolism, there is little information regarding their identification and distribution among native species from the rocky fields. Our results indicated a wide morphological diversity of reserve structures and sugar composition among the analyzed species and highlighted useful plants for future studies on ecophysiology and carbohydrate metabolism. Supported by CNPq, FAPESP and
PNADB/CAPES.
P0483 – ePoster
Recapturing rhizobial diversity in the Hunter Valley,
NSW, Australia
Fisher, N 1
Newman, K
, Cole, M
1
1
, Offler, C 1
, Nussbaumer, Y 1 , Dibley, K 1 ,
1 Centre For Sustainable Ecosystem Restoration,
University of Newcastle, Australia
Since European settlement the Hunter Valley has undergone huge levels of biotic loss. This includes soil
494 biota involved in many ecosystem services. Many of the losses are likely to be linked to the clearing of native vegetation, grazing and the subsequent loss of soil horizons due to erosion. Our research includes a focus of recapturing the soil microbes that remain and are involved in nitrogen supply to higher plants and their reintroduction as part of a process of reconstructing sustainable native ecosystems. This has been largely funded by the coal industry (Xstrata Coal and Thiess
P/L), but includes significant contributions from Federal
(ARC) and State (NSW) governments, with the latter funding research into the reconstruction of Ecologically
Endangered Communities.
The Mount Owen open cut coal-mining complex that encompasses the Ravensworth State Forest is a focus site for our research that is now listed as a ‘highly commended’ site on the Global Restoration Network of the Society for Ecosystem Restoration, International.
Thus, the outcomes of our research are freely available, globally.
Nitrogen has been found to be the most limiting plant nutrient in our studies in the Hunter Valley. Thus reintroducing the micro-organisms responsible for the majority of biological nitrogen fixation, the rhizobia, is crucial to the success of our ecosystem reconstruction efforts. As is common with many vegetation systems in
Australia, native legumes comprise a significant part of the flora. At the Ravensworth State Forest Vegetation
Complex, there have been 24 Fabaceae and 16
Mimosaceae species identified from vegetation surveys.
As part of the reconstruction process, systematic trapping of rhizobia has been undertaken to ascertain the diversity of the rhizobial population and to culture these bacteria for re-introduction into the re-constructed forest ecosystems. In addition, sampling of other sites in the
Hunter Valley has begun to maximize the recapture of the remaining rhizobia not lost to habitat destruction or competition with introduced agricultural strains. Not only have the expected genera of BradyRhizobium and
Rhizobium been found, but, purported native rhizobia bacteria have been isolated, including those from other genera. These include novel nodulating bacteria identified as belonging to the genus Burkholderia , thus confirming the recently discovered widespread occurrence of this genus within Australia. As well as the symbiotic nitrogen-fixers we have currently identified, bacteria thought to be endophytic root nodule occupants from the genera Burkholderia , Pseudomonas ,
Agrobacterium , Bacillus , Paenibacillus and
Brevibacillus that have been isolated from root nodules.
These root-nodule endophytes have implications for the complexity of root nodule occupation and pose questions as to how the plant organ (nodule) provides benefits to plant growth and contributions to the ecology of soil.
P0484 – ePoster
The role of new and old plant hormones in arbuscular mycorrhizal symbiosis
Foo, E 1
1 School of Plant Science, University of Tasmania,
Australia

The arbuscular mycorrhizal symbiosis between plants and Glomeromycota fungi is thought to be an ancient and widespread process, occurring in up to 80% of land plants. An important factor in the establishment of this symbiosis is communication between plant and fungi through the rhizosphere. Strigolactones, a group of terpernoid lactones, are one of the signals produced by the plant that promote the symbiosis in part by stimulating hyphal branching of the mycorrhizal fungi.
Recently, strigolactones were also discovered to act in planta as a suppressor of shoot branching and have been hailed as a group of new plant hormones. In addition to recently described strigolactones, many of the traditional plant hormones have also been proposed to play a role in the establishment of mycorrhizal symbiosis. Pea ( Pisum sativum L.) is a unique system in which to explore the role of both the new and old plant hormones in mycorrhizal symbiosis as a suite of mutants deficient in the synthesis and/or perception of strigolactones, auxin, gibberellin and brassinosteriods are available. A survey of the mycorrhizal status of these pea mutants will be presented, along with an exploration of the interaction between the traditional plant hormones and strigolactone biosynthesis, at a molecular and chemical level.
P0485 – ePoster
Exploring the interactions between gibberellin and strigolactones in branching control
Foo, E 1 , Reid, JB 1 , Ross, JJ 1
1 School of Plant Science, University of Tasmania,
Australia
Shoot branching is a key determinant of shoot architecture and has important consequences for plant adaptation and survival. Shoot branching is a tightly regulated process controlled by the classical plant hormones auxin and cytokinin as well as the recently discovered new group of plant hormones the strigolactones, which specifically inhibit bud outgrowth.
However, it has long been observed that many gibberellin-deficient dwarf mutants also display increased shoot branching. It has been speculated that this outgrowth of branches in dwarf shoots may be due to redirection of resources 'left over' from reduced stem elongation. Alternatively, it may indicate a more direct role for gibberellin (GA) in branching control. To further explore the role of GAs in branching control we have generated a new pea ( Pisum sativum L.) mutant line that is severely GA-deficient (na-1; GA biosynthesis mutant) but is partially compromised in its ability to synthesise growth-inhibitory DELLA proteins, which are destabilised by GAs. This new mutant line contains less bioactive GA than other GA mutants of similar height but branches much more profusely. These observations indicate that the branching phenotype of the new dwarf line is not due to altered resource allocation, pointing instead to a more direct role for GA in branching control.
We investigate if the increased branching of the new dwarf mutant is due to altered production of, or response to, the branching inhibitor strigolactone. We show that expression of PsRMS1, a key strigolactone biosynthesis gene, is increased in some GA-deficient lines and expression levels from the new dwarf line will be reported. The results from physio-chemical quantification of strigolactone production by dwarf mutants and wildtype roots will also be presented. In addition to investigating strigolactone level, potential changes in strigolactone response in the new dwarf line are explored using the synthetic strigolactone, GR24.
P0486 – ePoster cDNA-AFLP of
Psidium guajava
L. (guava plant) exposed to ozone
Furlan, C 1
FM 1
, Moraes, RM 2 , Santos, DYAC 1 , Rezende,
1 Universidade de São Paulo, Brazil;
Botânica São Paulo, Brazil
2 Instituto de
Plants are normally under biotic and abiotic stresses, such as drought, low nutrient availability, air pollutants and attack by pathogens and herbivores. Interaction of different stress factors results in synergism, affecting biochemical and physiological processes, with reflections in plant growth and development. Harmful effects derived from global climate changes possibly impart losses in crop productivity. Studies aiming to detect gene expression induced by stress factors may help evaluating the degrees of susceptibility of commercial cultivars, minimizing losses in productivity. This study aimed to examine the gene expression profiles, and secondary metabolites content of young individuals of a commercial cultivar of Psidium guajava L. (guava, Myrtaceae) subjected to ozone fumigation. P. guajava is cultivated in
South America and Central America, Africa and Asia for in natura uses or commercial production of jams, icecreams, beverages, juices, syrups and fruit pulp. Major guava producers are India, Mexico, Brazil and Malaysia.
Brazil produces yearly about 300,000 tons of guava fruits from the cultivars ‘Paluma’, ‘Pedro Santo’ and ‘Ogawa’.
Saplings of P. guajava cv. Pedro Sato were obtained from a Brazilian producer specializing in shoot production by rooting semi-herbaceous guava cuttings.
Eight saplings of each cultivar were kept one week on two treatments: filtrated air (FA) and ozone fumigated
(OF). At ozone treatment, AOT40 reached 566 ppp.h-1.
After this period, the leaves of the third node (counting from top to bottom) were collected, frozen in liquid nitrogen and stored (-80°C) to further analyses. cDNA-
AFLP analysis were conducted using the Plant Regular
Genome kit (Applied Biosystems). The 64 available combinations of EcoRI and MseI primers were tested, and the profiles were visualized on an automatic sequencer. Five combinations with higher number of polymorphic fragments were selected for cluster analysis and TDFs recovering from polyacrylamide electrophoresis gel. UPGMA analysis using Euclidean distances revealed the formation of one group composed by individuals undergoing stress. The same saplings were analyzed regarding their contents of anthocyanins, tannins and cuticular wax constituents, showing significant differences between treatments. Cluster analyses confirmed different expression profile between stress treatment and control, as well as, different contents of secondary metabolites. Sequence analyses of TDFs obtained in this work are still on course and will help to better understand expression profiles of a tropical species during ozone stress and their relationship with chemical defenses.
495

P0487 – ePoster
Physiological responses of suspension-cultured
Coryllus avellana
cells to low-intensity ultrasound
(US)
Ghanati, F 1 , Safari, M 1 , Abdolmaleki, P 2
1 Dept Plant Biol., Fac. Biol. Sci., Tarbiat Modares
University, Tehran, Iran; 2 Dept Biophysics, Fac. Biol.
Sci., Tarbiat Modares University, Iran
The application of ultrasound (US) to plant systems has opened a fascinating approach to plant physiology and biotechnology. The use of ultrasound to enhance the secondary metabolites biosynthesis in suspensioncultured cells has been reported frequently, mostly emphasizing on physical role of US on membranes resulting to the more extrusion of metabolites in the medium. However, the physiological role of US is yet to be elucidated. Taxol, an anticancer drug which is usually extracted from Taxus sp., has been isolated from
Coryllus avellana cells as well. The present study was undertaken in order to clarify the effect of low-intensity ultrasound (US) on cell growth, oxidative membrane damage, Taxol production in a rapidly growing cell line of Coryllus avellana cells. Six day old cells (in logarithmic growth phase), in LS media were exposed to
US at power density of 1.4 and 19.5 W for 2 to 40 min.
The US producing apparatus was locally designed and was able to emit modulating waves with the same range of authentic oscillation of the cells, after scanning with different frequencies. This caused the cells oscillated with the resonance range. The cells were then immediately returned to the fresh media and allowed to continue to growth. Growth parameters and protein,
MDA, H2O2 contents and Taxol production were evaluated. According to the results, exposure to 1.4W US up to 40 min had no significant effects on cell viability, growth, and membrane integrity, while increased the production of Taxol. Ultrasound with power of 19.5 W however, resulted in significant decreases in growth, viability, and protein content but increased MDA and
H2O2 contents of the cells in exposure periods longer than 4 min. Considering the fact that in both cases US resulted in more production of Taxol and more release of it to the media, from the results presented here it is more likely that applied US stimulated biosynthesis of Taxol, rather than merely damage of membranes and increase of extrusion of Taxol from the cells.
P0488 – ePoster
Effect of aluminum on the expression of gene and the activity of catalase in suspension-cultured cells of
Lisinthus
(
Eustoma grandiflorum
)
Ghanati, F 1
M 2
, Yousefzadeh Boroujeni, L 1 , Behmanesh,
1 Tarbiat Modares Univ., Iran; 2 Dept Genetics, Fac. Biol.
Sci., Tarbiat Modares Univ., Tehran, Iran
Exposure of plants to various environmental stresses including ultraviolet, pathogen infection, and toxic heavy metals can lead to over production of reactive oxygen species (ROS), so called oxidative stress. Aluminum is
496 the third abundant element in the earth which influences plant growth adversely in acidic soils. Although it is not a transition metal and does not catalyze redox reactions, the involvement of oxidative stress in Al toxicity has been proved. Among different ROS scavenging enzymes, catalase is likely the most important enzyme which detoxifies bulk of produced H
2
O
2
in Al-tolerant plants.
Lisianthus ( Eustoma grandiflorum ) is an ornamental plant which application of aluminum provides a longer vase life and increase water uptake by its cutting flowers.
The level of membrane lipid peroxidation, activity of catalase and expression of its gene was subjected to study in suspension-cultured lisinthus cells treated with or without different concentrations of Al. The cells were treated with 0, 200,400 and 800
µ
M of Al as AlCl
3
in a simple salt solution containing 3% sucrose and 0.2 mM
CaCl
2
, buffered with 2 mM Homo-pipes, pH 4.2. The cells were harvested at 1, 6, 12 and 24 h of the treatment.
Expression of catalase gene and activity of the enzyme increased in all concentrations of Al at all intervals, compared to those of the control cells. The rate of peroxidation of membrane lipids also reduced in Altreated cells in all concentrations and intervals. The results suggests that increase of vase life and increase of water uptake by lisianthus cutting flowers in the presence of Al, at least in part, is related to rapid, effective and long lived scavenging of hydrogen peroxide and the improvement of integrity of cell membranes.
P0492 – ePoster
Impact of salt stress on growth, nodule nitrogen and carbon fixation in the Desi (Indian) and Kabuli cultivars of chickpea
Gulati, R 1
1 DAV College Chandigarh, India
Four cultivars of chickpea, two of them of Mediterranean origin (Kabuli), CSG 9651, BG 267 and two Indian
(desi) types, CSG 8962, DCP 92-3, were compared for their relative salt tolerance in terms of growth, nitrogen fixing potential and carbon metabolism. The seeds of different cultivars were inoculated with Mesorhizobium ciceri, strain F: 75 and the plants were raised in the greenhouse.15- day old seedlings were administered doses of saline solutions of varying concentrations (0, 4,
6, and 8 dSm-1 NaCl, Na2SO4, CaCl2). Plants were harvested at 40, 70 and 100 days after sowing, for detailed analysis. Plant growth nodulation and nitrogenase activity (ARA) was more severely affected in
BG 267 and DCP 92-3 under salinity treatments (6 and 8 dSm-1) compared with CSG 9651 and CSG 8962. Root biomass was severely affected under salt stress. Although nodule number increased with salinity, further development of nodules was severely affected this was elicited in the form of reduced size as well as nodule weight. However CSG 9651 and CSG 8962 were able to tolerate stress and had significantly better ARA in their nodules as compared to other cultivars.
Phosphoenolpyruvate carboxylase (PEPCase) activity was higher in the nodules of tolerant cultivars under salt stress at all harvests. Salt stress limits nitrogen fixation and carbon metabolism capacity but the negative effects of salt stress were dependent on concentration and cultivar. The results indicated that significant variabilities

existed amongst different cultivars of chickpea for salt tolerance and the tolerant cultivars possessed an ability to grow and fix nitrogen in saline soils.
P0493 – ePoster
Redox chemistry of secondary metabolites: implications for evolution, producers and consumers
Hadacek, F 1 , Chobot, V 1
1 Dept of Chemical Ecology and Ecosystem Research,
Faculty of Life Sciences, University of Vienna, Austria
Oxygen is reduced to water by transfer of four electrons, a typical electron transfer process during respiration.
Incomplete transfer of those four electrons results in the formation of reactive oxygen species (ROS), including superoxide anion radical, hydrogen peroxide and hydroxyl radicals. Likewise, imbalance between photosystems I and II causes molecular oxygen to be reduced to superoxide anion radical. The hydroxyl radical is the most destructive of all ROS; its formation is determined by the transition metal-catalysed reduction of hydrogen peroxide that, again, arises by dismutation of superoxide anion radicals, either enzymatically
(superoxide dismutase) or non-enzymatically (a little bit slower). This scenario illustrates that 'wrong' electron transfers in the oxic world may cause the oxidation of biomolecules in living organisms. It is well known that secondary metabolites, especially phenols, can scavenge the free radicals and hydrogen peroxide by reducing them to water. Conversely, the same molecule, only depending on its redox potential and its chemical environment, can reduce molecular oxygen into superoxide anion radical, a pro-oxidative effect. This chemistry will be exemplified by juglone, a naphthoquinone that reputedly is viewed as the main causative agent for walnut tree allelopathy. It is feasible to assume, at least in an alternative paradigm, that the chemistry to interact with ROS may have contributed to the evolution of the diverse secondary metabolites. Yet they are produced by prokaryotic microbes. Generally, microbes are decomposers and metabolites that help to reduce molecular oxygen to obtain hydrogen peroxide might prove as beneficial. If this oxidative chemistry is triggered in living tissues of plants and animals, these microbes turn into pathogens; otherwise they contribute to litter decomposition in soils.
In plants, secondary metabolites are stored usually in specifically adapted compartments, such as lactifers, oil ducts, trichomes or idioblasts. The above described chemistry is facilitated more by a neutral pH of 7.4 (as found in the cytoplasm) than by the lower pH in the vacuole. In precautions to avoid reactions between secondary metabolites and oxygen in the cytoplasm, the secondary metabolites are stored in such compartments and only come into contact with the cytoplasm after tissue damage. The oxidation reactions then either disturb the redox homeostasis of the affected cells which may result in the expression of genes aiming to reinstate redox homeostasis – or, if the damage is caused by a herbivore, contribute to its deterrence. All classes of secondary metabolites potentially react with ROS. Only phenols, however, are able to scavenge them. The thus formed radicals of the secondary metabolites polymerize or stabilize as semiquinone or quinone structures that may be regenerated by reduction. The second scenario, however, applies only to efficient redox cyclers such as juglone. Terpenoids, by contrast, directly react with
ROS, and the reaction products undergo a complex follow-up chemistry. Consequently, for all drugs and vitamins and other secondary metabolites that we consume, a pre-receptor chemistry of secondary metabolites has to be expected and, in this chemistry, the functional groups are more important than threedimensional structures.
P0494 – ePoster
CAM induction by nutrient deficiency and drought stress in
Guzmania monostachia
, a C3-CAM facultativ bromeliad
Hamachi, L 1 , Pereira, PN 1 , Mioto, PT 1 , Mercier, H 1
1 University of São Paulo, Brazil
Crassulacean acid metabolism (CAM) is a mechanism of
CO
2
concentration at the site of RUBISCO that improves water use efficiency and, therefore, is especially important for plants living in environments characterized by constant or intermittent limitations in water supply, such as desertic and epiphytic niches, respectively. The tank epiphytic bromeliad Guzmania monostachia is known as being capable of switching from C
3
to CAM photosynthesis under drought stress, when the water supply in the tank in no longer available. As other tank bromeliads, the main source of nutrients for G. monostachia is probably provided by the tank solution; therefore, a drought stressed plant is also prone to be under nutritional deficiency. Moreover, it has been observed that tank bromeliads have some degree of longitudinal specialization along the leaf blade: the basal portion appears to play a major role in nutrition, whereas the apical portion is more associated with photosynthesis.
In this work our purpose is to present recent results relative to the influence of nutrient deficiency in CAM induction. The data obtained in our laboratory have indicated that not only drought stress, but also the nutritional deficiency is required to induce CAM in the leaves of this species. In plants of G. monostachia treated with a complete nutrient solution (Sarruge) and polyethylene glycol 6000 (PEG6000) 30% (m/m) solution (for drought stress), the drought stress alone was not able to induce CAM in the upper sections of leaves presenting a low level of nocturnal accumulation of acidity. Instead, it was also necessary a lack of nutrients.
Thus, we decided to carry out a succeeding experiment in which detached fully expanded leaves of G. monostachia were kept on the following treatments: (1) water, (2) 50% diluted Sarruge nutrient solution, (3) Sarruge solution,
(4) PEG6000 30% (m/m) solution, (5) PEG6000 30% solution plus 50% diluted Sarruge solution, and (6)
PEG6000 30% solution plus Sarruge solution and, subsequently, the degree of CAM in each treatment was determined. The degree of CAM was assessed by titratable acidity, and phosphoenolpyruvate carboxilase and Malate dehydrogenase activities. The correlation between nutrient deficiency and CAM induction by water stress will be discussed.
497

P0495 – ePoster
Comprehensive mechanisms for adaptation of barley to alkaline soil
Higuchi, K 1
1 Tokyo University of Agriculture, Japan
Nutritional disorders, such as micro essential element and
P deficiencies, and excessive NH4+ and OH- levels disturb the growth of plants in alkaline soil. Barley
( Hordeum vulgare ) is an alkaline soil-tolerant crop.
Under Fe deficiency, which is one of the often-observed nutritional disorders, the barley plant can solubilize the insoluble Fe in the soil by secreting mugineic acid phytosiderophore from its roots. This strong Fe acquisition mechanism has been well studied. In this study, we discovered several additional mechanisms that enable barley to grow on alkaline soil. 1. Photosystem I has a particularly high Fe requirement; therefore, Fe deficiency causes imbalance in electron flow and severe damage to the components of photosystems even under normal light conditions. We found that barley photosystems can induce thermal dissipation to compensate for excess excitation energy under Fe deficiency. Rice and sorghum, which are Fe-deficiency susceptible crops, cannot induce such a response under
Fe deficiency. Thus, barley photosystems possess specific mechanisms for adaptation to Fe deficiency [1].
2. The barley plant can protect its photosystems from severe damage caused by Fe deficiency, but the photosynthetic rate naturally decreases under Fe deficiency. The synthesis and secretion of mugineic acids involve the consumption of the assimilated C and N.
Therefore, the barley plant must optimize its metabolism to ensure the growth of newly emerged leaves and to acquire Fe from soil. We reported that Fe deficiency accelerates the senescence of old leaves in the barley plant [2]. The expression patterns of C, N, and S assimilation-related molecules in barley leaves and the concentrations of N compounds suggested retranslocation of N from Fe-deficient old leaves [3].
Moreover, the proportion of the leaf blade of the main culm to the whole shoot increased in Fe-deficient barley.
These adaptive mechanisms may contribute to the sustainable growth of barley under Fe deficiency. 3. Our results revealed that the primary disturbance in plant growth due to high pH is reduced root elongation [4].
This is a reasonable result because the elongation of plant cells is facilitated by acidification of the cell wall, which is known as the 'acid-growth theory'. However, we found that several plant species can elongate their roots at pH values greater than7.5. The barley plant could sufficiently elongate its roots in buffered nutrient solutions even at pH 9, but the rice and sorghum plants could not. We propose comprehensive mechanisms such as nutrition acquisition, nutrition assimilation, and architecture regulation for adaptation of barley to alkaline soil. [1] Saito et al. 2010. [2] Maruyama et al.
2005. [3]Higuchi et al. 2011 [4] Kobayashi et al. 2010.
P0496 – ePoster
What factors affect the induction of cell death in exponential- and stationary- Phase BY-2 Cells under still conditions?
498
1
Hiraga, A 1 , Kaneta, T 1 , Sato, Y 1 , Sato, S 1
Biology and Environmental Science, Graduate School of
Science, Ehime University, Matsuyama, Japan
Our earlier studies showed that still culture conditions induced cell death of tobacco BY-2 cells and this cell death was accompanied with apoptotic characteristics such as cell shrinkage, chromatin condensation, 'DNA laddering' pattern formed by multiples of about 200bp internucleosomal units, and TUNEL (transferasemediated dUTP nick end labeling)-positive.
Noteworthily, Evans Blue staining indicated that exponential-phase cells died more rapidly than stationary-phase cells when they were placed under still conditions. In the present study, we studied whether factors such as cell populations, conditions of the medium, and rates of the cell cycle progression affect the induction of cell death. The cells were very sensitive to the cell populations. In particular, exponential-phase cells died more rapidly than stationary-phase ones under still conditions when cell populations were increased. They were also sensitive to conditions of the medium. We examined the frequencies of dead cells when the cells were transferred into the fresh medium or the waste medium which were retrieved from the stationary-phase culture medium. When the cells were retrieved from the exponential- and the stationary-phase culture medium, respectively, and were transferred into the fresh medium, both of them did not die so rapidly under still conditions.
They showed only less than 30% of cell death even at the second day after the transfer. Interestingly, stationaryphase cells were not so sensitive to the waste medium compared with exponential-phase cells. While almost all exponential-phase cells died under still conditions at the second day after the transfer, stationary-phase cells showed about 40% of cell death. Thus, exponential-phase cells were very sensitive to circumferential factors such as cell populations and conditions of the medium compared with stationary-phase cells. About ten percent of exponential-phase cells are in the mitotic phase and mitotic cells are rare in the stationary-phase, so the accessibility of cell death under still conditions is expected to correlate with the cell cycle progression. So, we examined the frequencies of cell death when the cells were subjected to still conditions after the treatment with hydroxyurea (HU) for 24h. The result showed that the cells released from HU died rapidly compared with the control cells at least after 48h.
P0497 – ePoster
Minimum hydraulic safety leads to maximum water use efficiency in a forage grass
1
Holloway-Phillips, M-M 1 , Brodribb, T 2
Tasmanian Institute of Agricultural Research,
University of Tasmania, Hobart, Australia; 2 School of
Plant Science, University of Tasmania, Hobart, Australia
Understanding how water-use regulation relates to biomass accumulation is imperative for improving crop production in water-limited environments. Here we examine how the vulnerability of xylem to water-stress induced cavitation and the coordination between water transport capacity and assimilation influences diurnal

water use efficiency (WUE) and dry-matter production in
Lolium perenne L. – a commercial forage grass. Plants were exposed to a range of water stresses, causing up to
90% leaf death, by withholding water and then rewatering to observe the recovery process. Leaf hydraulic conductance (Kleaf) declined to 50% of maximum at a leaf water potential of -1MPa whereas complete stomatal closure occurred well after this point at -2.35MPa, providing no protection against hydraulic dysfunction.
Instantaneous assimilation remained maximal until >70% of hydraulic conductivity had been lost. Post-stress rewatering showed that 95% loss of Kleaf could be incurred before the recovery of gas exchange exceeded 1 day, with a rapid transition to leaf death after this point.
Plants exposed to sustained soil water deficit through restricted nightly watering regimes did not suffer cumulative losses in Kleaf, instead leaf water potential and gas exchange recovered diurnally. The effect was improved WUE during the day and optimal leaf water potential during the night for the maintenance of growth.
P0498 – ePoster
Long-term effects of sewage sludge land application on physiological parameters of
Asclepias syriaca
and heavy metal concentrations in soils and plant tissues
Huerta, AJ 1 , LeGalley, E 1
1 Miami University, Dept of Botany, Oxford, Ohio, USA
Application of heavy metal-containing sewage sludge to agricultural land as a fertilizer or soil amendment poses a risk to plants and other organisms that rely on the soil.
For eleven consecutive years beginning in 1977, sewage sludge was applied to plots of land at Miami University’s
Ecology Research Center in Oxford, Ohio. Short-term studies of these plots deemed the land application of sewage sludge as relatively safe. However, long-term follow-up studies that also consider plant physiological parameters are lacking. This project aimed to determine the long-term effects of sewage sludge land application on current heavy metal concentrations in sludge-treated soils, as well as the effects on several physiological parameters and heavy metal concentrations in the plant
Asclepias syriaca (common milkweed), twenty-two years after sewage sludge land application ended. Heavy metal concentrations in soil and plant tissue were measured by
ICP-MS. Heavy metal concentrations (specifically Cr,
Ni, Cu, Cd, and Pb) were significantly higher in sludgetreated soils and these concentrations varied significantly with depth. Current soil heavy metal concentrations are similar to those reported in earlier short-term studies, indicating that a very small proportion of these heavy metals leached away in the past 22 years. Photosynthetic and gas exchange parameters were measured with the
LICOR 1600 Steady State Porometer and the LICOR
6400 Portable Photosynthesis System. Leaves from plants of A. syriaca growing in sludge-treated soil showed significantly reduced stomatal conductance and transpiration rates and increased CO ff
compensation points. Analysis of heavy metal content in plant leaf tissue by ICP-MS showed that metal uptake by A. syriaca was similar to values reported from short term studies of other plant species 22 years ago. Furthermore, a significant difference in heavy metal uptake was found between leaves of different ages (e.g. Pb was concentrated in older leaves, Cd was concentrated in younger leaves). These findings may be of particular importance not only to environmental health, but also to human health, as sewage sludge is commonly applied to agricultural land were heavy metals pose the risk of bioaccumulating and entering the human food chain.
Heavy metal contamination has the potential to adversely affect the reproduction of plants and other organisms, thereby jeopardizing the broader ecological health of a region subjected to this environmental stress. The pursuit of potential remediation strategies for heavy-metal contaminated land is dependent upon continued monitoring of the health of organisms reliant on contaminated soils.
P0499 – ePoster
Tissue-specific distribution of calcium-bound pectin in cell wall during fruit development and ripening in tomato
Hyodo, H 1 , Furukawa, J 1 , Satoh, S 1 , Iwai, H 1
1 Graduate School of Life and Environmental Sciences
University of Tsukuba, Japan
Fruit ripening is a developmental process accompanied by seed development. This process of fleshy fruits involves many physiologiocal processes including the production of nutrients and aromatic compounds, changes in color, and softening of pericarp, which have evolved to attract animals and promote seed dispersal.
The molecular pathways that underlie many of ripeningrelated phenomena have been characterized for the modification of fruit nutritional and organoleptic status, and the role of ethylene in ripening, such as changes in color, flavor, and the shelf-life. However, the critical molecular determinants of fruit firmness and softening have not been well known. Fruit softening is a one of the most prominent parameter in fleshy or climacteric fruits.
For more than 40 years, many research have been targeted the mechanism of fruit softening, much of it using tomato fruit as a model system to study fleshy fruit development and ripening. A decline in fruit firmness occurs due to dissolution of the primary cell wall and middle lamella, resulting in a reduction in intercellular adhesion, depolymerization and solubilization of hemicellulosic and pectic cell wall polysaccharides.
These events are accompanied by increase expression of various cell wall degradation enzymes. For example, polygalacturonase(PG)-catalyzed depolymerization of pectin and pectin methylesterase (PME)-catalyzed demethylesterification of pectin in the wall and middle lamella have long been believed to be the principal process underlying fruit softening in tomato. Although it is well known that pectin depolymerization and modification is a remarkable change in tomato fruit ripening, and it does not affect fruit softening directly, the function of pectin during fruit ripening have not been well understood. Although many reports on tomato fruit ripening are focused on relation between pectin degradation and softening of whole fruit or pericarp, the changes in pectin content and/or composition during fruit ripeing might be unique by each tissue. In this study, to understand the tissue-specific roles of pectin during fruit development and ripening, we examined the pectin degradation and biosynthesis-related gene expression,
499

enzymatic activities, cell wall composition, and Ca content, involved in pectin synthesis and depolymerization in fruit. Our results show that changes in pectin property during fruit development and ripening have tissue-specific patterns.
P0500 – ePoster
Effect of cement particulates on eco-physiological behaviors of halophytes in the salt marshes of the Red
Sea, Saudi Arabia
Ibrahim, M 1 , Abdul Rahman, A 1
1 King Saud University, Saudi Arabia
The effect of cement dust on pigments status was studied for Zygophyllum coccineum , Salsola tetrandra , Cyperus conglomeratus , Limonium axillare and Suaeda vermiculata . Measurements were taken on plants closed to early constructed cement factory in the area (about 500 m) and other (about 4500–5000 m) far from the factory.
Cement dust increased the mortality of young branches leading to a reduction in the height and cover of the five species, especially Salsola tetrandra and Suaeda vermiculata , which were most sensitive to the dust. In most studied species, photosynthetic pigments including chl a, b and carotenoids were negatively correlated with dust deposition and positively correlated with the more distance from the factory. Photosynthetic pigments were declined faster with cement accumulation in Salsola tetrandra and Limonium axillare than in Zygophyllum coccineum and Suaeda vermiculata , whereas, increased significantly in Cyperus conglomerotus . Total carbohydrates and protein contents were decreased only with leaves of Salsola tetandra and Suaeda vermiculata, whereas, increased significantly near to the factory and parallel to dust accumulation in the other studied plants.
Cement dust pollution greatly affect the photosynthetic pigments of studied species, disturbing the pH of the cell sap and interrupted the metabolism of soluble amino acids and soluble sugars into insoluble reserve substances.
P0503 – ePoster
The chloroplast transcriptomes of C
4
Kajala, K 1 , Small, I 1
leaf
1 Plant Energy Biology, ARC Centre of Excellence, UWA
The two-celled C
4
photosynthetic cycle enhances the radiation use efficiency and growth rates of plants at high temperatures by concentrating CO
2
around RuBisCO
(Ribulose Bisphosphate Carboxylase / Oxygenase). The
C
4
cycle functions across two specialized cell types, mesophyll (M) and bundle sheath (BS) cells, each with distinct gene expression patterns and chloroplast differentiation. Although the biochemistry of these dimorphic C chloroplasts is understood, the regulation
4 of chloroplast gene expression in C
4
leaves has not been characterized. We have studied the chloroplast transcriptomes in Cleome leaves and characterized the changes required for C photosynthesis by two
4 comparative approaches. Firstly, the total leaf plastid transcripts of closely related C
4
C. gynandra and C
3
C.
500 spinosa have been compared by qRT-PCR to identify the changes involved in the evolution of C
4
photosynthesis.
Secondly, the M and BS cells of C. gynandra are being isolated with laser capture microdissection, and the plastid transcript levels of these cell type specific fractions will be compared by qRT-PCR, identifying the cell-type specific differences in chloroplast transcriptomes. The two complementary approaches provide novel information on the dimorphic chloroplast differentiation in the two cell types. Furthermore, we will identify potential chloroplast transcripts regulated in celltype specific manner via post-transcriptional regulation.
These insights will allow further work towards identification of the trans-factors regulating the cell-type specific chloroplast gene expression patterns.
P0505 – ePoster
Isolation of cold-regulated genes from garlic
Kim, N-S 1 , Kim, S-H 1 , Son, J-H 1 , Park, K-C 2 , Lee, S-I 1
1
2
Kangwon National University, Republic of Korea;
University of Seoul, Republic of Korea
Sessile plants are subject to many stresses from the fluctuating environment to have adapted internal mechanisms to tolerate to the external environmental stresses. Plants vary their ability to survive freezing temperature over the course of the year. Most temperate plants cold-acclimate, whereas tropical and subtropical plants are sensitive to chilling stress and largely lack the ability for cold acclimation. Garlic ( Allium sativum ) is an ancient crop of central Asian origin and has been cultivated in east Asia and east Europe for long. Garlic requires vernalization to stimulate sprouting and developing a bulb. Thus, cold tolerance is an important trait of garlic although the physiological depth of vernalization requirement varies depending on the varieties. While Southern varieties can hibernate in warm temperature, Northern varieties can tolerate even in chilling temperature. We have carried out a systematic analysis to isolate cold-regulated genes in garlic. We will present the molecular description of the cold-regulated genes in garlic.
P0507 – ePoster
Chemical diversity of the genus
Caesalpinia
(Fabaceae): new types of cassane diterpenes from
Caesalpinia crista
Kinoshita, T 1 , Shimada, M 1 , Narimatsu, S 1 , Jiang, D-X 1
1 Faculty of Pharmaceutical Sciences, Teikyo University,
Japan
The genus Caesalpinia is known to consist of more than
100 species. Though it constitutes a relatively large group in the family Fabaceae, chemical studies are limited to a few species among those. As far as results of chemical research undertaken so far are concerned, it seems that they are divided into two groups based on constituents: the one such as C. sappan is predominated by homoisoflavonoids, and the other such as C. bonduc by furano-cassane diterpenes. We conducted chemical investigation on C. bonduc of Southeast Asian origin and

reported the occurrence of a new type of hemiacetal cassanes fused with not a furan but an
α
,
β
-butenolide ring (5-hydroxyfuran-2(5H)-one), for which new names of neocaesalpin series are proposed. The previous chemical investigation on the same species that is exclusively of American origin revealed only the presence of furano-cassanes to which serial names of caesalpin are given. There will be two possible accounts for these results as follows. One is that an
α
,
β
-butenolide ring is artificial arising from addition of singlet oxygen on furan during either extraction or purification process, and the other is that those possessing an
α
,
β
-butenolide ring are not artificial but derived from a distinguished chemical race of C. bonduc which occurs widely in both the tropics and subtropics in the world. Our interest in cassanes featuring distinguished structural diversity instigated us to focus on other botanical sources. C. crista is one of four species of the genus Caesalpinia occurring in Japan, and chemical studies on the leaves of this species collected in the Rykyus furnished new neocaesalpin analogues named Neocaesalpins H and I.
However, those belonging to neocaesalpin group are obtained almost exclusively by our group. We are thus forced to consider the possibility of neocaesalpins being artificial, and carefully reinvestigated C. crista and C. bonduc using new plant materials collected in the same place. These studies furnished the isolation of two additional new compounds from the former species: one is a new type of nor-cassane fused with an ordinary furan, and the other is a 12-deoxy form of Neocaesalpin
H. The latter compound biosynthetically requires one more deoxygenation step that follows the possible oxygenation of a furan ring. Therefore, its isolation is of particular interest since it completely eliminates the possibility of neocaesalpins being artificial. Though the number of cassanes isolated from the corresponding
Caesalpinia species by our group is small, it revealed the distinguished chemical diversity based not only on the intra-specific differentiation of C. bonduc but also on the inter-specific differentiation among the genus
Caesalpinia , indicating that they are the promising source of new types of chemicals. During a search of phytochemical literatures concerning Caesalpinia species, the confused use of botanical names has often been encountered. What are referred to as C. bonduc and
C. crista in our study is C. bonduc (L.) Roxb. emend.
Dandy et Exell and C. crista L. emend. Dandy et Exell, respectively.
P0508 – ePoster
Changes of Chl a fluorescence of five hydrophytes under cadmium stress
Koh, SC 1 , Oh, SJ 1
1 Department of Biology, Jeju National University, Jeju,
Korea; 2Jeju Institute of Field Science Education and
Research, Jeju, Korea
The effects of Cd2+ ions on the Chl a fluorescence of five hydrophytes, Lemna paucicostata, Salvinia natans,
Ricciocarpus natans, Nymphaea tetragona, and Typha orientalis, were investigated in order to get informations on physiological responses of plants to Cd2+ stress and to select Cd2+-sensitive plant species. Lemna plants were most sensitive to Cd2+ stress, while Nymph plants were most tolerant. In all Cd2+-treated plants except for
Nymph plants, Fv/Fm, the maximum photochemical efficiency of PSII, decreased in proportion to the increase of Cd2+ concentration and treatment time. The Chl a fluorescence O-J-I-P transients were also changed considerably by Cd2+ ions; the fluorescence yield decreased considerably in steps J, I and P in Cd2+treated plants, although it followed a typical polyphasic rise in non-treated plants. In Lemna plants, the functional parameters, ABS/CS, TRo/CS, ETo/CS and RC/CS, decreased in proportion to the increase of Cd2+ concentration, while N, Mo and Kn increased. The structural parameters,
Φ po,
Φ po/(1-
Φ po), PIabs, SFIabs,
Kp and RC/ABS, also decreased according to the increase of Cd2+ concentration. These results suggest that several functional or structural parameters could be applied to determine quantitatively the physiological states of plants under stresses and Lemna plants could be useful as a experimental model system to investigate responses of plants.
P0509 – Poster
Plant litter degradation by freshwater fungi under heavy metal stress
Krauss, G-J 1 , Wesenberg, D 1 , Gudrun, K 2
1 Martin-Luther-University Halle-Wittenberg, Germany;
2 Helmholtz Centre for Environmental Research-UFZ
Leipzig-Halle, Germany
The activities of aquatic fungi (AQH) as decomposers of plant derived organic matter establish them as crucial components of stream biota (1). Research on freshwater fungi has concentrated on their role in plant litter decomposition. Fungal mycelia perform core ecosystem functions such as regulating the rate of litter mass loss, providing food for leaf shredders (accumulation of biomass) and for filter-feeders (production of conidia and fine leaf particles), and releasing inorganic nutrients.
Freshwater fungal diversity and activity is controlled by the environmental impact. The ability of AQH to withstand or mitigate anthropogenic stresses is becoming increasingly important. Metal avoidance and tolerance in
AQH implicate a sophisticated network of mechanisms involving external and intracellular detoxification (1).
Studies on fungal growth of exposed alder leaves in heavy metal polluted streams in Central Germany showed a slowly leaf decomposition in high polluted sites and fungal colonization was restricted to external surfaces. The leaf surface quickly became covered with crystalline deposits (zincowoodwardite) (2). Cd exposed
AQH show a specific thiol peptide response (1). Heliscus lugdunensis seems to be the first organism described, which under Cd stress synthesize a novel small metallothionein together with a phytochelatin,and accompanied by increased glutathione levels (1).
Examining adaptive responses in plant litter degradation under metal stress will unravel the dynamics of biochemical processes and their ecological consequences.
(1) Krauss, G.-J. et al. (2011). (2) Ehrman, J.M. et al.
(2008).
501

P0511 – ePoster
Autoclavable and low temperature operative Cu/Zn superoxide dismutase
Kumar, A
G 1
1 , Bhardwaj, PK 1
, Ahuja, PS 1 , Kumar, S 1
, Sahoo, R 1 , Dutt, S 1 , Bagler,
1 Biotechnology Division, Institute of Himalayan
Bioresource Technology (CSIR), Palampur, Himachal
Pradesh, India
Superoxide dismutase (SOD; EC 1.15.1.1) has immense applications in cosmetic, food, pharmaceutical, medical and plant industry with an estimated market potential of
$10 to $20 billion. Keeping in view the wide applications and scope, SODs with a wide range of kinetic stability are required for targeted or specialized applications.
From a high altitude plant Potentilla atrosanguinea , a novel Cu/Zn SOD (Pot-SOD) was identified that could be autoclaved with catalytic activity down to sub-zero temperature. The X-ray crystallographic structure of Pot-
SOD showed its existence as a dimer consisting of molecules A and B. Each subunit of Pot-SOD structure consisted of eight stranded β- sandwich (1a-8h) connected by seven loops (I-VII), one short helix and one
310 helix. Significant structural differences between molecules A and B were limited to two regions: the
Greek-key loop VI and the electrostatic loop VII. Single disulfide linkage found in SODs was conserved in Pot-
SOD. This S-S bond links loop IV and β-strand 8h and this linkage of the secondary â€'structure elements contributes to the stabilization of the SOD fold. Highly thermostable nature of Pot-SOD was assigned to increased number of hydrogen bonds and decreased gap volume of 2787.69 compared to other reported SOD.
Also, absence of conserved water molecule due to replacement of conserved Serine10 to glycine makes Pot-
SOD very stable. Further, kinetic studies revealed Pot-
SOD to have resistance to sodium dodecyl sulfate, stability to proteolysis, and pH storage stability in the range of pH 3 to 10. Pot-SOD was found to be resistant towards thermal denaturation. Circular dichroismspectroscopic analysis revealed that Pot-SOD consisted of high percentage of β-sheets content in agreement with most of eukaryotic Cu/Zn SODs, and retained secondary structure and activity after being autoclaved. Pot-SOD displayed a definitive monomer to dimer ratio.
Computational analysis with residue interaction graph model of Pot-SOD structure, that encapsulates longrange noncovalent interactions, complemented and supported the results. In order to widen the scope, flora was screened for SODs with unique kinetic properties.
SOD gene from corresponding plants were cloned and analyzed through heterologous expression system in E. coli . Apart from industrial applications of SOD, outcome of the research has enormous implications in protein engineering and bioeconomy.
P0512 – ePoster
Thionin phylogenies – what can we add to chemosystematics
Larsson, S 1
1 Div. of Pharmacognosy, Uppsala University, Sweden
502
Thionins are cysteine-rich proteins of about 50 amino acids, believed to be involved in plant defense against fungi and bacteria. Ever since their discoveries round the middle of the 20th century, as 'yeast lethal factor' in cereal flour and potent toxins in mistletoes, they have enticed scientists from fields such as plant breeding, medicinal chemistry and biotechnology.
Chemosystematic information is a well-known principle of using plant classification to increase 'hit-rate' in drug discovery. The more closely related two plants are the higher the possibilities for finding similar activity due to derivatives from a shared precursor compound. Using similar chemical compounds as classification tools for plants should be the natural quid pro quo of this. But assessing homology between low-molecular compounds usually needs knowledge about their biosynthesis, a field still mostly based on assumptions of extrapolation of data from a few model organisms. As genetically encoded compounds thionins conveniently lend their nucleic or amino acid sequences to phylogenetic analysis, and their size and remarkable stability simplifies their isolation.
However, just as low-molecular substances, they occur as cocktails of closely related compounds within species or genera. So which are the lessons we can learn from them in future use of chemosystematics for finding new drugs and other applications for use of chemical characters?
Differences in the number of cysteines (six or eight) in the sequences between thionin isoforms have been used to delimit subfamilies, but neighbour joining as well as parsimony analyses of the amino acid sequences fail to recognise such divisions – instead mainly grouping the proteins in clusters corresponding to the organisms they have been isolated from. Utilising data from the expressed preproproteins, mRNA or even the few published genes strengthens this latter result. Tracing the selectivity between thionins lytic activity in vitro on different types of cells also show a pattern where strong activity against bacteria and fungi is coupled to weak activity against mammalian cells (general trend for tested thionins from grasses), while strong cytolytic effects in screenings using mammalian cancer cell-lines is coupled to low activity against pathogenic bacteria and fungi (as seen with the thionins from mistletoes). The isoform cocktail found in bread wheat consists of three proteins, each have been shown to be derived from a single copy gene located within one of the original three genomes in this hexaploid. This is contrasted by the cocktail in diploid mistletoes that may contain seven thionins or more. So what is the lesson? That the assessment of homology is key to all chemosystematic comparisons and investigations.
P0513 – ePoster
Isolation and partial purification of alpha amylase
1 inhibitor from
Adiantum
fronds
Laware, S 1 , Limaye, A 1 , Deore, G
Fergusson College, Pune, India
1 , Shinde, B 1
Fresh fronds of Adiantum aethiopicum L. were collected at various growth stages like a) young fronds, b) mature fronds without sori, c) mature fronds with immature sori
(pale green colour) and d) mature fronds with mature sori stage (brown to black colour). They were extracted in
0.15M NaCl. The clear supernatant obtained after

centrifugation was subjected to protein fractionation with ammonium sulfate. The protein fractions were dialyzed against 0.15M NaCl and tested for salivary alpha amylase inhibitor assay. Fractions between 30 and 60% saturation (F30-60) proved to have maximum salivary amylase inhibitory activity; hence it was applied to a
Sephadex G-100 column and eluted at a flow rate of 3.0 ml min -10 . Each eluted fraction was measured spectrophotometrically at 280 nm and tested for salivary alpha amylase inhibitor assay. The elution number 6, 7,
8, 15, 16, 17, and 18 exhibited maximum salivary alpha amylase inhibitory activity. Maximum inhibitory activity
(68%) was observed in eluted fraction 8. The fraction is moderately thermo-stable as it is stable to the temperature ranging from 30°C to 60°C. The maximum activity was recorded at 35°C and retained about 60% activity up to 50°C. It was also proved to be stable to different pH ranging from 3 to 8, but at pH 7.0 it showed maximum inhibitory activity. The fraction was tested for its proteolytic actions against proteolytic enzymes such as trypsin and chymotrypsin. It was found to be more effective amylase inhibitor than the commercial antdiabetic tablet (Glucobay). Thus, the results of present investigation could suggest the use of alpha amylase inhibitor extracted from Adiantum aethiopicum can be used as hypoglycemic agent in non-insulin dependent diabetes mellitus and obesity patients.
P0514 – ePoster
VIVIPAROUS 1 interacting Protein 2, VIP2 is involved in mitogen-activated protein kinase signaling in rice plants
Lee, S-K 1
J 1
, Choi, Y-S 1
, Kwon, T-R 1
, Kim, S-Y 1
, Byun, M-O
, Kim, B-G
1 , Park, S-C 1
1 , Jeong, M-
1 National Academy of Agricultural Science, Republic of
Korea
Mitogen-activated protein kinase (MAPK) signaling cascades are known to be correlated with environmental, developmental and hormonal responses in higher plants.
Stress-responsive rice MAPK, OsMAPK5 has been revealed that it was induced by abscisic acid as well as pathogen infection and abiotic stresses showing inverse regulation in biotic and abiotic stress responses. In this study, we isolated putative up/downstream components by using yeast two hybrid system to know more about
OsMAPK5 signaling cascades. A putative transcription factor, VIVIPAROUS 1 Interacting Protein 2 (VIP2) was included as a positive candidate of OsMAPK5 interactors. Predicted MAP kinase phosphoacceptor recognition sequence Ser/Thr-Pro was presented in its coding region. It contained RING-finger (Really
Interesting New Gene) domain which is conserved in many proteins involved in the ubiquitination pathway for protein turnover. To confirm their interaction, recombinant GST-OsMAPK5 and His-tagged OsVIP2 were produced in E. coli . In vitro assay of protein kinase showed that OsMAPK5 retained autophosphorylation activity and was able to phosphorylate VIP2 in the preference of Mn
2
+ . These results suggest that OsVIP2 is a transcription factor that is directly phosphorylated by
OsMAPK5.
P0515 – ePoster
Cell wall polysaccharides and non-host resistance in barley
Little, A 1 , Burton, RA 2 , Schweizer, P 3 , Fincher, GB 2
1 School of Agriculture, Food and Wine, University of
Adelaide, Australia; 2 ARC Centre of Excellence in Plant
Cell Walls, University of Adelaide, Australia; 3 Leibniz
Institute of Plant Genetics and Crop Plant Research
(IPK), Germany
The interaction between a plant host and a fungal pathogen is complex, so in order to understand the overall relationship it is important to investigate individual components separately. In plants, the cell walls are the first line of defence which protects the cell from successful invasion. The pathogen and plant cell go into battle to see who has the strongest cell wall and most advanced structural and chemical weaponry. This leads to the question of what is the best way to build a plant cell wall in order to provide the broadest defence against fungal penetration. In order to answer this question we must look at the genes involved in constructing the building blocks of the plant cell wall. Plant cell walls are diverse structures consisting of many different cellulosic and non-cellulosic polysaccharides encoded by a range of glycosyltransferases. Microarray analysis of barley infected with the biotrophic pathogen Blumeria gramanis has identified a number of Cellulose synthase-like genes that are differentially expressed during the interaction
(Zellerhoff et al. 2010). Transient silencing of these genes has resulted in altered susceptibility and confirmed the role of cell wall polysaccharides in pathogen defence.
Stable barley transformants are available for the candidate genes and further analysis of the cell wall polysaccharide composition is underway.
P0516 – ePoster
1
Biogeography, habitat preference and morphology of
C
4
grasses: a phylogenetic perspective
Liu, H 1 , Edwards, E 2 , Freckleton, R 1 , Osborne, C
University of Sheffield, Sheffield, UK;
1
2 Brown
University, Rhode Island, USA
C
4
grasses comprise about 46% of around 10, 000 species in the grass family Poaceae, occupy 1/3 of the terrestrial area and show huge diversity in response to climatic factors like temperature and precipitation.
Understanding this diversity is crucial for grassland management, especially in of the context of anthropogenic climate change. Contrasting biogeographical relationships with mean annual rainfall have been reported for different three photosynthetic subtypes of C lineages of C
4
4
grasses. However, different evolutionary
grasses show even stronger relationships with rainfall than photosynthetic subtypes. This raises two questions: 1) whether a phylogenetic or physiological classification is most suitable for explaining C
4
grass distributions; and 2) whether these patterns are driven by water availability per se, or by correlated ecological factors like productivity or shading.
We analyzed morphological and habitat information at
503

the genus level for two monophyletic subfamilies,
Chloridoideae and Panicoideae, which together account for 90% of C
4
grass species. Phylogenetic groups associate tightly with photosynthetic subtypes, and model comparisons implied that phylogeny was the most important determinant of variation in morphology and habitat. Using phylogenetic trees based on our dataset,
Pagel’s lambda tests and phylogenetic independent contrasts (PICs) were carried out to investigate how genus morphological traits and habitat preference vary or co-vary across the phylogenetic tree. There were phylogenetic signals in both morphology and habitat, and
PICs showed that these signals arose mainly from the divergence between Chloridoideae and Panicoideae.
Genera of Chloridoideae had significantly shorter culm heights, narrower leaf widths and smaller guard cell lengths than those of Panicoideae, and they occupied drier habitats and were less likely to exist in shaded habitats. A phylogenetic signal in the relationship between plant traits and habitat occupancy revealed the indivisibility of phylogeny and habitat preference.
Therefore, although we found phylogenetically independent covariation of leaf width and plant size, the association of plant size with habitat water availability was phylogenetically dependent. Our results indicated that phylogenetic effects on plant size and habitat preference both play roles in the differential sorting of grass lineages along precipitation gradients. The phylogenetic background of species is therefore crucial for understanding biogeographic distribution patterns in relation to ecological factors.
P0517 – Poster
Effects of Chitosan on seed germination and physiological characteristics of safflower seedling under water stress
Mahdavi, B
M 1
1 , Modarres Sanavy, SAM
, Sharifi, M 1 , Khalesro, S 1
1 , Aghaalikhani,
1 Tarbiat Modares University, Iran
The effects of chitosan solutions on seed germination and physiological changes of safflower ( Carthamus tinctorius
L.) seedling were investigated under water stress conditions. Treatments comprised of factorial combinations of four osmotic potential levels (0, -4, -8 and -12 bar, prepared by PEG 6000) and nine chitosan concentrations (0, 0.05, 0.1, 0.2, 0.4, 0.5, 1, 2, 3%, all dissolved in 1% acetic acid) along with a additional treatment of distilled water. Result showed that germination percentage, germination index, shoot and root height, shoot and root dry weight and protein content decreased with increasing water stress. Proline and malondialdehyde (MDA) content increased by increasing osmotic potential to -8 bar. The greatest shoot height and dry weight (19.3 and 36% more than distilled water treatment, respectively) were observed at 0.4% chitosan concentration. At low concentrations of chitosan
(0.05–0.4%) protein and proline content increased and
MDA content decreased. In osmotic potential -8 bar, the highest germination percentage, shoot and root height obtained by 0.4% chitosan concentration. They had no significant difference with distilled water and acid acetic
(0% chitosan). Also at this osmotic potential the decline
MDA and proline content and the increase of the
504 concentrations of protein (in comparison with distilled water treatment) were detected in low concentrations of chitosan. In the highest level of osmotic potential (-12 bar), germination percentage, shoot and root height increased by low concentrations of chitosan in comparison with distilled water treatment. Also at this osmotic potential the least rate of former parameters obtained by 1-3% chitosan. Thus, it suggests that chitosan concentration at 0.4% or lower levels may improve germination percentage of safflower seeds and benefit for seedlings growth under water deficit stress.
P0518 – ePoster
Hepatoprotective property of
Cassia sophera
Linn. on carbon tetrachloride (CCl
4 rats
)-induced hepatotoxicity in
Maity, TK 1 , Satish, M 1 , Singh, J 1
1 Dept of Pharmaceutical Technology, Jadavpur
University, Kolkata, India
Methods: Chronic hepatotoxicity was induced by CCl
4 intoxication in rats. Serum biochemical parameters and histopathological studies were carried out with methanol extract of Cassia sophera leaves using methodology of
Malloy & Evelyn (1937) and Kind & King (1965).
Results: Rhamnetin (O-methylated flavonol), one of the active component present in this plant species, was isolated from the aerial parts by silica gel column chromatography employing gradient elution with chloroform: methanol solvent mixture. Characterization of Rhamnetin was done by mp, UV-Visible spectrophotometry, chemical analysis, FT-IR, 1H NMR,
13C NMR and mass spectral analysis. The protective effect of Cassia sophera extract (MECS) was studied on
CCl
4
-induced toxicity in rats. Post-treatment with methanol extract of Cassia sophera (200 & 400 mg/kg body weight) after CCl administration significantly
4 prevented the elevated serum enzymatic activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and total bilirubin. The total serum protein level was increased to normal level as it was decreased with CCl
4 intoxication. Histopathological changes induced by CCl
4 were also significantly attenuated by MECS treatment in both preventive and curative experiments. Conclusions:
The plant extract has a potent hepatoprotective effect on
CCl
4
-induced liver injury in rats.
P0519 – ePoster
A study on the hepatoprotective activity of ethanol extract of
Bacopa monnieri
L. against ethanol-induced
1 hepatotoxicity in rats
Maity, TK 2 , Gosh, T 1 , Singh, J 2
Dept of Pharmacy, Contai Polytechnic, Darua, Purba
Medinipur, India; 2 Dept of Pharmaceutical Technology,
Jadavpur University, Kolkata, India
Oxidants have been shown to be involved in alcohol induced liver injury. This study was designed to test the hypothesis that ethanolic extract of Bacopa monnieri aerial parts (EBM) protects against early ethanol-induced

liver injury in rats. Results show that EBM was effective in blunting ethanol-induced enhanced activities of
SGOT, SGPT and alkaline phosphatase. EBM was also effective in reducing ethanol induced increase of bilirubin and cholesterol in blood. EBM was also effective in arresting ethanol-induced increase in production of malondialdehyde and reduced glutathione
(GSH). Furthermore, EBM could blunt ethanol-induced suppressed activities of superoxide dismutase (SOD) and catalase (CAT). EBM was also effective in reducing ethanol-induced lipid peroxidation in vitro. EBM also could blunt body weight loss because of ethanol. This study demonstrates that EBM prevents ethanol toxicity developed by ethanol presumably by blunting oxidative stress. Results of hepatocellular damage caused by ethanol and its recovery by EBM, suggest that EBM might be considered as a potential source of natural hepatoprotective agent. The hepatoprotective activity of
Bacopa monnieri L. extract could be related to the free radical scavenging properties of saponins present in high concentration in the extract.
P0520 – Poster
Nitrate reductase activity in pineapple plants submitted to low temperatures in different phases of light/dark cycle
Matsumura, AT 1 , Mercier, H 1
1 Universidade de São Paulo, Brazil
Nitrate reductase (NR), a key enzyme responsible for reducing nitrate to nitrite, is widely known to exhibit a marked daily rhythm of activity, with maximum values at begging of light phase and minimum levels in dark phase. Previous studies carried out at our laboratory have shown that pineapple plants, when grown under thermoperiod of 25°C light/15°C dark for 3 months, exhibited highest NR activities in roots during the dark phase, while plants grown in constant temperature of
28ºC displayed preferentially NR activities in leaves mainly during the light phase. The aim of this study was to analyze the interaction between light and low temperatures on NR activity in leaves and roots of pineapple plants in different phases of the light/dark cycle. Pineapple plants were obtained by micropropagation technique and grown in vitro for 3 months in a constant temperature of 25ºC, photoperiod of
12 hours and 50 µmoles photons m -2 s -1 . In the first set of experiments, plants were submitted to 10ºC or 25°C
(control group) at middle of the light phase for 1, 3, 6 or
9 hours, in the presence or absence of light. The results showed that, in the presence of light, there was a positive influence of low temperatures in leaf NR activity, whilst in the absence of light there was a positive influence of low temperatures in root NR activity. Subsequently, the influence of phase (light/ dark cycle) was tested on NR activity at low temperatures. Plants were exposed at 10ºC or 25°C (control group) for 6 hours at middle of light phase or middle of dark phase, and NR activity was measured after that every 3 hours throughout 24 hours in rewarming treament (25ºC). Results indicated that when plants were exposed to low temperature at middle of light phase, leaves showed an increase of NR activity after 3 hours of rewarming (dark phase), while in the control group, NR activity decreased. In roots, NR activity had an increment after 6 hours of rewarming (dark phase), but no other significant changes were observed. When plants were subjected to low temperature at middle of dark phase, leaf NR activity reached its maximum near beginning of dark phase, after 9 hours of rewarming, while in the control group, the maximum activity was observed at begging of light phase, resulting in 6 hours of delay in NR daily cycle. In roots, there was an increment of activity after exposure to low temperature, but following rewarming, no significant changes of behavior were observed. All these results suggested that low temperatures can trigger distinct responses in NR activity of leaves and roots, dependent on the presence or absence of light. When the effects of rewarming after low temperature exposition were tested, it revealed that results were different according to the phase of light/dark cycle. Altogether, it indicates that low temperature delays the NR daily cycle in leaves, corroborating results obtained in other plant species. Nevertheless, the effects of low temperature on roots will need more experiments for further conclusions.
P0521 – Poster
Low night temperature influences positively the activity of nitrate reductase associated with plasmatic
1 membrane in roots of pineapple plants
Souza, A 1 , Matsumura, AT 1 , Mercier, H 1
Universidade de São Paulo, Brazil
Nitrate reductase (NR) is responsible for reducing nitrate to nitrite, usually described as a citossolic enzyme (NRc).
However, some works point to the existence of a plasmatic membrane form (NRPM), with distinct regulatory ways. Previous work carried out at our laboratory have showed that pineapple plants, when grown under thermoperiod of 28°C light/15°C dark for 3 months, exhibited the crassulacean acid metabolism
(CAM) and highest NR activity during dark phase in roots, while plants grown in constant temperature of
28°C, exhibited the C
3
photosynthesis and highest NR activity during the light phase in leaves. CAM plants are caracterized by temporal separation between CO
2
uptake, which occurs at dark phase and its decarboxylation that occurs during light phase. It was believed that noctunal uptake of CO
2
could provide reducing power or carbon skeletons for nitrate reducing, explaining the high NR activity found in dark phase. Initially, the aim of this study was to analyze the influence of CAM photosynthesis on the high nocturnal NR activity found in previous work, using pineapple plants induced to
CAM by high light intensity. The plants were obtained by micropropagation technique and grown in vitro in a constant temperature of 25ºC, photoperiod of 12 hours and 50 µmoles photons m -2 s to 200 µmoles photons m -2
-1 . They were then submitted
s -1 in the same conditions, while control remained at low light intensity of 50
µmoles photons m -2 s -1 . After 30 days, plants cultivated in higher light intensity presented CAM photosynthesis, pointed by nocturnal acidity accumulation and malate dehydrogenase activity. The in vivo NR activity was measured every 3 hours throughout 24 hours in CAM and C
3
plants (control). Surprisingly, the results did not show higher NR activity at dark phase in CAM plants
(roots and leaves) when compared to control, suggesting
505

that the high root NR activity, which was found in previous work was due to nocturnal low temperature.
Therefore we decided to test the influence of the low temperature on different types of NR. The fractionation method was standardized for pineapple plants and citosolic and membrane fractions obtained were submitted to the NR in vitro assay. Plants were submitted to thermoperiod of 28°C light/15°C dark for 30 days, when was observed high nocturnal root NR activity, and plants grown under constant temperature of 28°C were used as control group. The method described above was applied only to roots. According to the results, in citosolic fractions there were not significant differences in NR activity when compared to control group.
However, in membrane fractions there was a higher NR activity in roots submitted to thermoperiod of 28°C light/15°C dark, demonstrating that the differences verified in total root NR activity were due to the increase of NRPM activity. In conclusion, this work suggests that pineapple plants present both forms of NR in roots (NRc and NRPM), with distinct ways of regulation. It seems that low night temperature can trigger root plasmatic membrane form and the induction of CAM has no influence on the rise of NRPM activity.
P0522 – ePoster
Analysis of overexpressor of MIZU-KUSSEI1, a gene required for root hydrotropism of
Arabidopsis thaliana
Miyazawa, Y 1 , Moriwaki, T
, Takahashi, H 1
1 , Uchida, M 1
A 1 , Fujii, N 1
1 Dept of Life Sci., Tohoku University, Japan
, Kobayashi,
All organisms depend upon water availability. Unlike mobile organisms, land plants are sessile in nature and must complete their life cycles where they germinate.
Accordingly, land plants have evolved various mechanisms for responding to unfavorable environmental signals, allowing them to tolerate or avoid environmental stresses such as water scarcity. To avoid drought conditions, roots sense environmental stimuli and respond by regulating growth away from dry areas or toward wet areas. To regulate growth in a directional manner, land plants have evolved tropisms, whereby organ growth is redirected in response to an environmental stimulus, such as gravity, light, a moisture gradient, or touch. Indeed, plant roots can sense a moisture gradient and grow toward the wet area, a process termed positive hydrotropism. Recently, we established experimental system for the induction of hydrotropic responses in Arabidopsis roots, which had enabled us to use molecular genetics as a tool. Genetic screens based on the inability to develop hydrotropic root curvature allowed us to isolate a series of ahydrotropic mutants termed 'mizu-kussei (miz)'. Recently, we succeeded in identifying the genes required for hydrotropism, namely MIZ1 and MIZ2 / GNOM, which were the ahydrotropic mutants whose responsible genes had ever been determined. In contrast to MIZ2 / GNOM, which encodes an ARF-GEF, the function MIZ1 is unknown. Physiological and morphological analyses of the mutants showed that miz1-1 had normal gravitropic responses, root elongation growth and root cap organization, while it completely lacked hydrotropic
506 response. MIZ1 encodes a protein of unknown function with a conserved domain at its C-terminus, which we termed MIZ domain. The genes encoding MIZ domain are found only in the genomes of land plants, and the
MIZ domain has no clear similarity to any characterized peptide sequence. To analyze the physiological roles of
MIZ1, we generated transgenic plants that express the
GFP-fused MIZ1 in addition to MIZ1 overexpressing plants. The fluorescent signal of GFP-fused MIZ1 was expressed at the root tip, especially in the lateral root cap cells and cortical cells. On the other hand, the signal of
GFP-fused miz1-1 was scarcely detected. Since the mRNA levels of MIZ1 were equivalent between wild type and miz1-1, it is suggested that the missense mutation in miz1-1 caused instability of the corresponding gene product. Furthermore, we found that overexpression of MIZ1 conferred an enhanced hydrotropism. This enhancement in hydrotropism was completely nullified by either miz2 mutation or brefeldin
A treatment, suggesting that MIZ1 functions upstream of
MIZ2. As the sensory apparatus for hydrostimulation is considered to reside at the root tip, our results suggests that MIZ1 positively regulates hydrotropism at its early phase.
P0523 – ePoster
Analysis of cytochrome P450 from
Polygonum
1 tinctorium
Mizuta, Y 1 , Imada, Y 1 , Hayase, M 1 , Minami, Y 1
Okayama Universtiy of Science, Okayama, Japan
In Japan, Polygonum tinctorium is traditionally grown to produce a natural blue dye indigo. The leaves contain a large amount of indican, a colorless precursor of indigo.
Indican degradation enzyme,
β
-glucosidase, is also present in the leaf cells. If the leaves are ground, indican, indoxyl-
β
-D-glucoside, would be degraded by
β
glucosidase to indoxyl and glucose. Because indoxyl is an unstable compound, it is immediately oxidized to indigo by oxygen present in the air. Indican degradation enzyme is located in chloroplasts, while the substrate indican is only stored in the vacuoles. Therefore, insoluble indigo is produced, only when the leaf cells are physically broken: for example, by invasive insects. We anticipate that the physiological role of indican storage in cells may be a defense system against predators. Our purpose is to elucidate the total synthetic pathway of indican and its role in vivo. To date, we have purified and characterized an indican synthase, which is an UDPglucosyl transferase. This enzyme catalyzes the synthetic reaction of indican using indoxyl and UDP-glucose as substrates. The optimum pH of the reaction (pH 10) indicates that the enzyme cannot function in the vacuole where the final product indican is stored. Furthermore, it is thought that the precursor of indoxyl synthesis is indole, an intermediate in the synthetic pathway of tryptophan. We suppose that the hydroxylation of indole is catalyzed by cytochrome P450. In many plants, cytochrome P450s and UDP-glucosyl transferases in the endoplasmic reticulum (ER) relate to various secondary metabolisms. Because indoxyl is a very unstable compound, indoxyl synthesized by the catalysis of cytochrome P450 needs to conjugate immediately with glucose. Therefore, the two enzymes of indican synthesis

may coexist in ER. Here, we report on the cDNA cloning, expression, and characterization of cytochrome
P450 from Polygonum tinctorium . cDNA of cytochrome
P450 was cloned from the cDNA library of Polygonum leaf tissue. The isolated cDNA consisted of 1,732 bp encoding a polypeptide of 507 amino acids. The Nterminus has a signal peptide consisting of 40 amino acids. The primary structure also conserves several motifs specific to cytochrome P450, such as membrane spanning, oxygen-binding, electron transfer protein binding, and heme-binding regions. Polygonum enzyme, which we named ptCYP1, was compared with cytochrome P450s from various plants. The ptCYP1 sequence shows the highest identity (36%) to CYP83B1 that uses indole alkaloid as substrate. In addition, it shows 35% identity to CYP71C9v1 that acts in the oxidation of indole. Therefore, we assume that ptCYP1 is also related to indole metabolism. To examine whether ptCYP1 can oxidize indole into indoxyl, the recombinant protein was expressed in E. coli . We also report the characteristics of the recombinant ptCYP1.
P0524 – ePoster
1
Environmental regulation of stomatal response in
Arabidopsis
Cvi-0 ecotype
Monda, K 1 , Negi, J
Hashimoto, M 1
1 , Iio, A 2 , Kusumi, K 1
, Sakakibara, H 3 , Iba, K 1
, Kojima, M 3
Dept of Biology, Kyushu University, Japan; 2
,
National
3
Institute for Environmental Studies (NIES), Japan;
RIKEN Plant Science Center, Japan
Genetic approach using mutant varieties is a powerful tool for understanding differences in stress sensitivity among plants. However, this approach usually focuses on only one target gene at a time. On the contrary, environmental stress tolerance involves a large number of genes at the same time. Genetic variation between naturally occurring Arabidopsis populations is large, providing a unique source to study the effect of complex genetic variation on stress tolerance, for example, natural genetic variation in response to water deficit. The
Arabidopsis Cape Verde Islands (Cvi-0) ecotype is known to have a low tolerance to drought stress. There is a possibility that this phenotype is closely related to stomatal response. In this study, we investigated the stomatal behavior of Cvi-0 plants in response to environmental signals. Leaf temperature provides a convenient indicator of transpiration and can be used to detect phenotypes with altered stomatal control.
Compared to the most popular Arabidopsis ecotype Col-
0, high [CO
2
]-induced increase in leaf temperature was inhibited in Cvi-0, showing that stomatal closing response was restricted. We also investigated the response of stomatal conductance in Cvi-0 and Col-0 to changes in the CO
2
content in the air. Cvi-0 showed constitutively higher stomatal conductance than Col-0.
This result was consistent with the above Cvi-0 phenotype of leaf temperature. The measurement of stomatal conductance in Cvi-0 to light and dark exposures with ambient [CO
2
] showed that Cvi-0 retained the ability to respond to light and exhibits stomatal opening, but the responsiveness was slow.
Epidermal peels experiments showed that Cvi-0 was defective in regulation of transpiration in response to drought stress. Under low-humidity conditions, stomata of Cvi-0 were more opened compared to those of Col-0.
Abscisic Acid (ABA) is a regulatory molecule involved in drought stress tolerance and enhances stomatal closing response. We used epidermal peels to examine ABAinduced stomatal closure. Cvi-0 stomatal response exhibited a strong insensitivity to ABA. We then assessed whether low humidity affects endogenous ABA levels in Col-0 and Cvi-0. Under low-humidity conditions, Cvi-0 had much higher ABA levels than Col-
0. These results suggested that stomata opening phenotype of Cvi-0 was due to ABA insensitivity.
Measurements of organic and inorganic ions in guard cell protoplasts indicated an over-accumulation of the osmoregulatory anions (malate and Cl-) in the cells of
Cvi-0. This result may provide clues to explaining the characteristics of environmental responses of stomata of the Cvi-0 ecotype.
P0529 – ePoster
Optimization of the cultivation conditions for the employment of key factors driving to maximization of extracellular production of
Nostoc strains from the paddy fields of Iran
Nowruzi,
F 1
1. 5 , Khavari-Nejad, RA
, Nejadsattari, T 2
3 , Kazemi, B 4 , Najafi,
1 Dept of Biology, Faculty of Science, Teacher Training
University, Tehran, Iran; 2 Dept of Biology, Faculty of
3 Dept of Science, Islamic Azad University, Tehran, Iran;
Biotechnology, Shahid Beheshti University of Medical
4
5
Sciences, Tehran, Iran; Dept of Parasitology, Shahid
Beheshti University of Medical Sciences, Tehran, Iran;
Cellular and Molecular Biology Research Center,
Shahid Beheshti University of Medical Sciences, Tehran,
Iran
Many cyanobacteria are known to be able to synthesise outermost slimy investments and to release polysaccharidic material into the culture medium during cell growth. Even though the presence of polysaccharidic investments is a quite conserved feature among cyanobacteria, a great lack of knowledge exists on the structure-/function relationships of cyanobacterial EPS as well as on the factors that regulate EPS biosynthesis. The cyanobacterial genus Nostoc polysaccharides are attracting increasing interest for their potential applications in the pharmaceutical industries, competing with other natural polysaccharides obtained from plants and macroalgae. Different environmental, nutritional and physical parameters affected production and accumulation of the EPS. The aim of this study is establishment of the key culture factors driving to the maximization of EPS production and preserving noncapsulated morphotypes of Nostoc strains in the culture media, which is crucial importance for the development of biotechnological applications. Actually, the presence of thick polysaccharidic investments imposes serious difficulties for the extraction of bioactive substances and for the molecular biology studies of these strains. In the present study, two strains of Nostoc sp. FSN and Nostoc sp. ASN were cultured in diazotrophic (BG-110 medium) and non-diazotrophic (BG-11 medium) conditions in the presence of urea, nitrate and ammonium as nitrogen sources. After adaptation, the strains were cultured at
507

both continuous light (180µEm -2 s -1 ) and light/dark (16/8 h) cycles from 30–35°C for about 1 month in aerated batch cultures. Non-capsulated and capsulated strains of
Nostoc were collected from the paddy fields of Iran. The cultures were sampled daily and chlorophyll a and cell dry weight were determined. After 30 days of cultivation, protein and cell density were measured by biophotometer instrument and extracellular polysaccharide and released polysaccharides were quantified spectrophotometrically by the phenol/sulfuric acid method. Continuous light enhanced total carbohydrate synthesis in the noncapsulated strain ( Nostoc sp. FSN) in diazotrophic and non-diazotrophic cultures and the type of nitrogen metabolism did not affect significantly the amount of total carbohydrate recorded. In the presence of nitrate, urea and ammonium, the capsulated strain ( Nostoc sp.
ASN) became non-capsulated. Long period of hormogonia and aseriate colony in this strain led to increasing the viscosity of the aqueous solutions and amount of EPS was higher than that of non-capsulated strain ( Nostoc sp. FSN) under the light/dark (16/8 h) cycles. Since cyanobacterial extracellular polysaccharides are interesting from the biotechnological point of view, considering many other parameters are of crucial importance for the maximization of EPS production in this genus.
P0530 – ePoster
On the presence of peptide synthetase and polyketide synthase genes in the cyanobacterial genus
Nostoc as a mining tool for secondary metabolites
Nowruzi, B 1. 5 , Khavari-Nejad, RA 3 , Kazemi, B 4 , Najafi,
F 1 , Nejadsattari, T 2
1 1Dept of Biology, Faculty of Science, Teacher Training
2 University, Tehran, Iran; 2Dept of Biology, Faculty of
Science, Islamic Azad University, Tehran, Iran; 3 3Dept of Biotechnology, Shahid Beheshti University of Medical
Sciences, Tehran, Iran; 4 4Dept of Parasitology, Shahid
5
Beheshti University of Medical Sciences, Tehran, Iran;
5Cellular and Molecular Biology Research Center,
Shahid Beheshti University of Medical Sciences, Tehran,
Iran
Cyanobacteria produce a tremendous variety of secondary metabolites. Many of these compounds have biological activities, making cyanobacteia an excellent source of new drug candidates. Approximately 40% of cyanobcateria secondary metabolites are cytotoxic and many others have antibiotic, antifungal or antiviral activities. These products are peptides or depsipeptides.
They have features such as incorporation of nonproteinogenic amino acids which are characteristics of peptides biosynthesized by non-ribosomal peptide synthetases. Some of these peptides also have acetate- derived moieties, suggesting that their biosynthesis also involves polyketide synthases. In the present study, it was focused on the detection of nonribosomal peptide synthesis and polyketide synthase genes in two stains of
Nostoc sp. FSN and Nostoc sp. ASN, presenting different capsular polysaccharidic morphologies and also released polysaccharide productions is different. These stains are the most frequent species of paddy fields in Iran. After 3 weeks growth, total genomic DNA was extracted from exponential phase cultures. The DNA sequences of the
508
16s rRNA region were determined to classify the Nostoc stains. Amplification of peptide synthetase and polyketide synthase genes were achieved by use of degenerative primers directed to conserve functional motifs of these modular enzyme complexes. The primers target the KS and A domains, respectively. All the products were screened using 1% agarose gel electrophoresis. Results showed amplification of the appropriate ~1-kb for peptide synthetase and ~700-bp fragments for polyketide synthase. Also, analyses of antibiogram bioassay were screened. In vitro antibacterial activity of organic extracts of Nostoc strains were evaluated against Gram-positive bacteria
(Staphylococcus aureus) and Gram-negative bacteria
(Escherichia coli, Pseudomonas aeruginosa). Dried extracts and supernatants were dissolved in several solvent (methanol, hexane, ethanol, petroleum ether, ethyl acetate) and antimicrobial activity was determined by the disc method. The ethyl acetate extract of Nostoc sp. FSN showed more potent activity against
Staphylococcus aureus than that of other organic extracts.
However, culture supernatant found to be inactive against all the test organisms. No inhibitory effect was found against Escherichia coli, Pseudomonas aeruginosa.
Gram-positive bacteria were found to be more susceptible as compared to Gram-negative bacteria.
According to these results, it is concluded that the antibiogram bioassay and molecular detection of peptide synthetase and polyketide synthase genes in Nostoc may be useful techniques for the assessment of natural product -producing species and possible role of peptide synthetase and polyketide synthase enzyme complexes in the biosynthesis of biologically active compounds
P0531 – ePoster
Analysis of lignescens, a novel temperature-sensitive
1 mutant of
Arabidopsis
exhibiting ectopic lignin deposition
Nozaki, M 1 , Sugiyama, M 2 , Sato, Y 1
Biology and Environmental Science, Graduate School of
2
Science, Ehime University, Matsuyama, Japan;
Botanical Gardens, Graduate School of Science,
University of Tokyo, Bunyo-ku, Japan
Lignin, a complex phenylpropanoid polymer, functions in development and defense in vascular plants. Lignin deposition occurs in vascular tissues and in wounded or pathogen-infected sites. We isolated a novel temperaturesensitive mutant of Arabidopsis that deposits a large amount of lignin ectopically, and designated it as lignecens (lig) . To obtain new insights into the regulatory mechanisms of lignin biosynthesis, we have performed phenotypic and molecular characterization of the lig mutant. The lig seedlings grown at the restrictive temperature (28°C) showed severe growth defect associated with ectopic lignin deposition in the stele of the root, which was clearly observed by the phloroglucinol staining. When the lig seedlings were exposed to 28°C after cultured at the permissive temperature (18°C), they ceased root growth within 24 hours and accumulated lignin in the stele of the primary root tip within 32 hours. These observations indicate that the growth defect of the lig mutant is not secondary result of the lignin deposition. By chromosome mapping and

sequence analysis, a missense mutation possibly responsible for the lig phenotype was found in AtGNA1 , which encodes glucosamine-6-phosphate acetyltransferase (GNA1). The phenotypes of lig at 28°C were complemented by the introduction of the wild-type
AtGNA1 gene, leading to the conclusion that the LIG gene is AtGNA1 . The lig mutant version of the AtGNA1 protein (AtGNA1 G68S) produced in E. coli was purified and examined for the activity to acetylate glucosamine-6phosphate. The enzyme activity of AtGNA1 G68S was stable at 18C but rapidly disappeared at 28°C. GNA1 is known to be involved in the biosynthesis of UDP-
GlcNAc, a donor of GlcNAc for the synthesis of N linked and O -linked glycans. Therefore it can be reasonably speculated that the growth defect and the ectopic lignin deposition in the lig mutant may result from the deficiency of UDP-GlcNAc. To test this hypothesis, we examined effects of administration of
UDP-GlcNAc on the phenotypes of the lig mutant. In the presence of 1 mM UDP-GlcNAc, the root growth of lig at 28°C was recovered to approximately 80% of control, and the ectopic deposition of lignin was suppressed.
Additionally, the wild-type plants treated with 100 nM tunicamycin, an inhibitor of the first step of N-glycan synthesis, i.e., transfer of GlcNAc-1-phosphate from
UDP-GlcNAc to dolichol phosphate, exhibited lig -like phenotypes, including growth defect and ectopic lignin deposition in the stele near the root tip. These results suggest that the deficiency of UDP-GlcNAc can cause ectopic lignin deposition as well as growth defect through the repression of N -glycan synthesis.
P0532 – ePoster
The chemical ecology and defense profiles of
Australian mistletoes
O'Gorman, KL 1 , An, M 2 , Gleadow, RM 1 , Miller, RE 1 ,
Watson, DM 3
1 School of Biological Sciences, Monash University,
2 Clayton, Australia; Environmental and Analytical
Laboratories, Charles Sturt University, Wagga Wagga,
Australia; 3 Institute for Land, Water and Society,
Charles Sturt University, Albury, Australia
Mistletoes are shrubby hemi-parasites which obtain all their water and nutrients through a cellular connection to their hosts’ xylem. The dissolved nutrients are passively transported to the mistletoe as a result of its typically high transpiration rate. Subsequently, mistletoe foliage is highly nutritious and relatively succulent; this makes it a popular food choice for browsing mammals and herbivorous insects. Plants defend their tissues with a range of secondary compounds and structural defenses. It is unlikely that a plant will be defended with a single trait and instead will invest in a range of characteristics known as defense syndromes. Mistletoes rarely invest in structural defenses but a wide range of secondary compounds have been found in the tissues of species that belong to Viscaceae and in some from Loranthaceae.
This study represents the first systematic study of plant secondary compounds in Australian mistletoes and is one of few studies to examine the chemical ecology of parasitic plants generally. By evaluating variation in the chemical profiles of mistletoe species from various genera in the Loranthaceae family, the relative importance of parasite identity and host-plant identity will be determined, and the evolutionary basis of interspecific differences will be explored. Mistletoe chemistry has been known to vary according to identity of the hostplant and in some cases mistletoes have been shown to directly sequester secondary compounds from their hosts.
With the exception of Tasmania, mistletoes in the
Loranthaceae family are found in all major areas and climates in Australia. Collectively, they parasitise a broad range of both native and exotic species and whilst some mistletoe species have a low host-specificity, others are almost exclusively found on one species of host. In addition to identifying secondary compounds in mistletoe species, this study aims to determine if there are any differences in the defense syndromes of closely related host-species, of both infected and non-infected individuals.
P0533 – ePoster
A lectin with alpha-galactosidase activity from cowpea hypocotyls possessed a yieldin activity to regulate the yield threshold of cell wall
Okamoto-Nakazato, A 1 , Naito, C 1
1 Showa Pharmaceutical University, Japan
Extension of plant cell walls that are driven by turgor is defined by the mechanical properties of cell wall such as extensibility (f) and yield threshold (y). In the acidgrowth process induced by auxin, the acidification of cell walls results in an increase of f and a decrease of y, then, causes a drastic extension. The acid-induced shift of y is regulated by yieldin, a protein which is isolated from the cell walls of the elongating region on the cowpea hypocotyls on 2000. The molecular cloning of yieldin reveals the fact that yieldin is a homologue of concanavalin B, a kind of lectin. However, the mechanism of yieldin to decrease y still remains uncertain.To study the action mechanism of yieldin, its lectin activity was examined with hemagglutinin tests using trypsinated rabbit red blood cells. Yieldin, the
30kD-protein isolated from the cowpea hypocotyls with an affinity chromatography using anti-yieldin antibodies, showed a hemmagglutinin activity (HA-activity) under pH 7, but the HA-activity of yieldin was lower than that of the other 25kD-protein which was also recognized by the anti-yieldin antibody. SDS-PAGE and gel filtration analysis indicated that the 25kD-protein, tentatively called cowpea lectin-like protein (CLP), was approximately 100 and 25kD under pH 7 and 4 respectively. These results imply that the monomer of
CLP, which is assumed to share a homologous or the same protein structure as yieldin, acts as an
α
galactosidase under pH 4, and makes a homo-tetramer that acts as a lectin under pH 7. The yieldin activity of
CLP to change y under acidic conditions was also examined. The yield threshold of the glycerinated hollow cylinder (GHC) of the cowpea hypocotyl was ca. 120 gw under pH 7 and shifted to ca. 75 gw under pH 4. The acid-induced shift of y was extinguished by heattreatment on the GHC, and was recovered by the reconstruction with CLP in the similar extent by yieldin.
This experiment showed that CLP also possessed yieldinactivity. The effect of inhibitors on the lectin/
α
galactosidase -activity of CLP and on the acid-induced
509

shift of y was also investigated. The activity of CLP as a lectin was inhibited by 0.02 mM of galactose and 5 mg/ml of hydroxyl-mercuribenzoic acid (HBA), but not by deoxy-galacto-nojirimycin (DGN). The
α
galactosidase activity of CLP was inhibited by DGN and
HBA at lower concentrations (0.1
µ g/ml and 0.01 mg/ml respectively), and galactose at higher concentrations
(0.2M) than on CLP as a lectin. Meanwhile, DGN and galactose, at the same concentration that they inhibited the
α
-galactosidase activity of CLP, extinguished the acid-induced shift of y. However, HBA increased the acid-induced shift of y at the same concentration that inhibited CLP as a lectin. These results indicate that CLP as an
α
-galactosidase decreases y under pH4, and the
CLP as lectin potentially keeps y not to be decreased excessively. Thus, it is assumed that the switching ability of CLP from
α
-galactosidase to lectin in correlation with the pH plays an important role in the regulatory mechanism of y.
P0534 – ePoster
Vegetation geography of alpine-subalpine regions on
Japanese mountains viewed from vegetation of maritime–oceanic regions of northeastern Asia
Okitsu, S 1
1 Graduate School of Horticulture, Chiba Univ., Japan
Alpine–subalpine regions of Japanese mountains contain diverse vegetation types such as Abies mariesii forest,
Betula ermanii forest, Pinus pumila scrub and snow-bed herbs. They develop under per-humid climate which is characteristics among Japanese mountains. This study focuses on the vegetation geography of those vegetation types with a comparison of vegetation of maritime– oceanic regions of northeastern Asia. The aims of the study are: 1) to understand the floral composition and geographical distribution of the vegetation types, and 2) to reconstruct vegetation history of alpine–-subalpine regions of Japanese mountains. Subalpine region of
Japanese mountains consists of two sub-regions: tall tree region and shrub region. Major vegetation types of the tall tree region are: Abies mariesii forest, Betula ermanii forest, broadleaved herbaceous community. Major vegetation types of the shurb region are: Pinus pumila scrub, snow-bed dwarf shrub, wind-exposed dwarf shrub, alpine desert. Among them Abies mariesii forest is endemic vegetation to Japan, while others have ecologically relevant vegetation of maritime–oceanic regions of northeastern Asia. Betula ermanii forest exactly corresponds to that of the Kamchatka peninsula.
Broad-leaved herbaceous community has originated from understory of B. ermanii forest in the Kamchatka. Pinus pumila scrub can be considered to a southern expansion of understory of Larix gmelinii forest in Eastern Siberia.
Snow-bed dwarf shrub and wind-exposed dwarf shrub both are relevant vegetation to those of dwarf heath vegetation in northern Kuriles. Geographical distribution of the major components of those vegetation types are: 5 of Okhotsk type, 3 of East Siberian type, 2 of Bering type and 1 of circum polar type. This finding suggests that vegetation of alpine–subalpine regions of Japanese mountains develops under humid–per-humid climate of the northern Pacific. Subalpine vegetation of Japanese mountains in the Last Glacial contained also humid–per-
510 humid components, and offered them refugia, in contrast with the generally accepted view that in the Last Glacial
Japanese mountains experienced cold-dry continental climate and had continental arid vegetation types.
P0535 – ePoster
Differences among plant species in the rootelongation response to high pH
1
Ono, K
T 1
1 , Ito, T 1
, Miwa, E 1
, Ose, N 1 , Kawaguchi, Y 1
, Higuchi, K 1
Tokyo University of Agriculture, Japan
, Nishinosono,
Acidic conditions are required for cell elongation in the plant cell wall. This is known as the 'acid-growth theory'.
If this theory holds true for all plant species, then root elongation should be inhibited by high pH in all plant species. However, the optimum pH for root elongation in the Chenopodiaceae plant Suaeda salsa that grows on sodic soil ranges from 6 to 8. The objective of this study is to assess the variation among plant species with respect to the optimum pH for root elongation and to elucidate what kind of biochemical and physiological properties are responsible for these differences. Intact plant experiment: To compare the optimum pH for root elongation in intact plants from different species, we selected 16 plant species that had different tolerance levels to salt. We measured taproot elongation in the plants for 48 hours in buffer solutions with pH values of
5.0, 6.0, 7.5, or 10.0. The taproot elongation for
Arabidopsis thaliana , Thellungiella halophila , Spinacia oleracea , Beta vulgaris ssp. vulgaris , Sorghum bicolor ,
Solanum lycopersicum , Glebionis coronaria , Cucurbita maxima , and Glycine max was maximum at pH 5.0 or pH
6.0 and was greatly inhibited at pH 10.0. The optimum pH for Suaeda salsa , Kochia scoparia , Hordeum vulgare , and Solanum melongena was 7.5, and the inhibition of taproot elongation at pH 10.0 was lesser than that for other species. The taproot elongation in
Oryza sativa , Luctuca sativa , and Cucumis sativus was maximum at pH 6.0 and remained at 90% of the maximum level at pH 5.0 and pH 7.5. This rootelongation response to pH differed among the plant species. These differing responses may reflect the different pH values of the soil on which they have evolved. Root tip experiment: The elongation of intact roots was not enhanced in acidic solution in the case of
Suaeda salsa , K. scoparia, H. vulgare, and Solanum melongena . However, the acid-growth theory has been investigated in previous studies by using excised plant segments. Therefore, we assessed root tip elongation in the Chenopodiaceae plant K. scoparia and the Poaceae plant H. vulgare for which elongation of intact roots was maximum at neutral or alkaline pH. We used the
Chenopodiaceae plants Spinacia oleracea and the
Poaceae plant O. sativa for comparison. We measured root tip elongation for 1.5 hours in buffer solution at pH values of 5.0, 6.0, 7.5, and 10.0. The root tip elongation for all species was maximum at pH 5.0 or pH 6.0 and was greatly inhibited at pH 10.0. Therefore, we think that the acid-growth theory also holds true for each cell from plants where the elongation of intact roots is maximum at neutral or alkaline pH values. These results suggest that root cell elongation in intact alkaline-tolerant plants is due to not only acidic conditions in the cell wall but also

some factors from tissues other than those of the root tip.
In future studies, we will investigate whether signals from the shoots regulate root elongation under neutral and alkaline pH conditions.
P0536 – ePoster
Evaluation of callogenesis and rootgenesis of cotyledone and hypocotyl explants in
Brassica napus
L. under in vitro salt stress
Pakmehr, Y 1 , Razavizadeh, R 1 , Karamian, R
1 Payame Noor University, Iran
1
The genus Brassica L. belongs to Crucifereae family and comprises 5 species in Iran. Brassica napus L. is one of cultivated species of the genus, which are used for production of plant oil, animal food and biological fuels.
After palm and soybean it is the third cultivated crop, which is used for oil production in the world. In this research seeds of Brassica napus L. were cultured in MS medium containing different concentrations (0, 50, 100 and 150 mM) of NaCl. In addition, for study of the effect of NaCl on in vitro organogenesis and callogenesis, cotyledone and hypocotyl explants obtained from seedlings were cultured in MS medium containing different NaCl concentrations and also different concentrations of NAA and kinetin. Results showed the lowest frequency of callogenesis and rootgenesis at 150 mM NaCl. Moreover, the highest frequency of callogenesis and rootgenesis was obtained in saltless medium (control). Also other results from tissue culture experiments showed that the highest frequency of root induction was obtained in higher concentration of NAA and without kinetin and the lowest frequency of root induction was obtained in higher concentration of kinetin and lower concentration of NAA. The highest frequency of callus induction was observed in MS medium supplemented with 1 mg/l of NAA and 2 mg/l kinetin and the lowest was observed in MS medium supplemented with different concentration of kinetin and without NAA. The frequency of callogenesis and rootgenesis of hypocotyl explants was more than cotyledone explants.
P0539 – ePoster
Effects of seasonal variations on essential oil composition in
Rosmarinus officinalis
from Iran
Pazoki, A 1 , Faezbakhsh, A 2
1
2
Islamic Azad University of Varamin Brach, Iran;
Islamic Azad University, Central Tehran branch,
Tehran, Iran
The essential oil composition of the aerial parts of
Rosmarinus officinalis . (Lamiaceae) grown in Varamin .
Parts of plants were harvested from the same location at different times of the year ( 2009) and dried in a dark place so that the continuous air stream flows through.
The essential oil was obtained by hydrodistillation and has been analyzed by GC/MS and GC.The minimum and maximum amounts of camphor,
β
-pinene and other major compounds was same as Spanish rosemary and other studies. The main compounds that varied seasonally included
β
-pinene (highest in summer
16.44%), camphene (highest in summer: 9.21% ),
α
myrcene (highest in summer 8.06 ) 1,8-cineole (highest in summer 9.90%), borneol (highest in winter 16.87%) and comphor (highest in winter 12.57%). The percent of
1,8-cineole was almost constant throughout the year.
P0540 – ePoster
Phenotyping of
Brachypodium distachyon
to accelerate plant biofuel breeding.
1
Poiré, R 1 , Chochois, V 2 , Sirault, X 1
Furbank, R 1
, Vogel, J 3 , Watt, M 2 ,
High Resolution Plant Phenomics Centre, CSIRO Plant
Industry, Canberra, Australia;
Canberra, Australia; 3
2 CSIRO Plant Industry,
USDA, ARS, WRRC Albany,
California, USA
The High Resolution Plant Phenomics Centre (HRPPC) is the Canberra node of the Australian Plant Phenomics
Facility, located at CSIRO Plant Industry and the
Australian National University. The HRPPC focuses on deep phenotyping and reverse phenomics through development of next generation tools to measure performance of plants ranging from model species to major crops. A major aim of the HRPPC model plant module is to find genes of agricultural importance using high resolution phenotyping of model plant species. One such project is an international collaboration to develop
Brachypodium distachyon as a model cereal plant to speed up the breeding of next generation biofuel crops and find genes responsible for important traits in wheat.
Brachypodium, unlike switchgrass or miscanthus, has all the attributes of a model species (small and fully sequenced genome, short life cycle, easily transformed) allowing high throughput screening of traits relevant to biofuel research. We have developed a variety of noninvasive and destructive assays for growth, biomass, photosynthesis and root growth in Brachypodium . Large phenotypic variability of biomass accumulation (up to 5 fold difference between the top 25% and low 25% groups) was observed in a set of 160 natural accessions of Brachypodium . Root growth and architecture was also characterised in these ecotypes and show significant variation in term of total root length, root type distribution or shoot/root ratio. Contrasting ecotypes are currently under investigation using high resolution imaging analysis techniques to identify the optimal combination of these phenotypic traits and to determine the underlying genomic regions responsible, by high throughput re-sequencing techniques. Since
Brachypodium is a typical grass and shares a high degree of genetic similarity with other crops, it will be possible to extend the knowledge gained from this project to improve plants species such as switchgrass and wheat.
P0541 – ePoster
A graphical user interface for the analysis of vein and areole dimensions in leaves: software overview and preliminary scaling analyses
Price, C 1
511

1 School of Plant Biology, Unversity of Western Australia,
Australia
Interest in the structure and function of physical biological networks has spurred the development of a number of theoretical models that predict optimal network structures across a broad array of taxonomic groups, from mammals to plants. In many cases, direct tests of predicted networks are impossible given the lack of suitable empirical methods to quantify physical network structure with sufficient scope and resolution.
There is a long history of empirical methods to quantify the network structure of plants, from roots, to xylem networks in shoots and within leaves. However, with few exceptions, current methods emphasize the analysis of portions of, rather than entire networks. Here, we introduce the Leaf Extraction and Analysis Framework
Graphical User Interface (LEAF GUI), a user-assisted software tool that facilitates improved empirical understanding of leaf network structure. LEAF GUI takes images of leaves where veins have been enhanced relative to the background, and following a series of interactive thresholding and cleaning steps, returns a suite of statistics and information on the structure of leaf venation networks and areoles. Metrics include the size, position and connectivity of all network veins, and the size, shape and position of the areoles they surround.
Available for free download at www.leafgui.org, the
LEAF GUI software promises to facilitate improved understanding of the adaptive and ecological significance of leaf vein network structure. Preliminary Scaling
Analyses The LEAF GUI software was used to analyze interspecific scaling patterns from a global dataset of 353 leaves representing 353 species. Our results suggest that the scaling of leaf network morphology is driven by a characteristic length scale likely determined by diffusion limits between veins and stomata. This manifests in several invariant quantities such as mean vein length, mean distance to the nearest vein, whole leaf network density. Moreover, in many respects the scaling of leaf networks is indistinguishable from a null model of an isometric lattice due to the numerical dominance of the highest order veins. However, network scaling differs from the lattice model in important ways, particularly with respect to the frequency distributions of vein lengths and diameters which were consistent with exponential and a power law models respectively.
P0542 – ePoster
Morpho-anatomical and physiological responses of
Centaurea ragusina
L. to salinity
Radic Brkanac, S 1
H 2
, Glavas, K
, Pevalek-Kozlina, B 1
1 , Peharec, P 1 , Lepedus,
1 University of Zagreb, Croatia;
Osijek, Croatia
2 Agricultural Institute
To elucidate the physiological basis of salt tolerance in the Croatian plant species Centaurea ragusina L., the accumulation of salt ions and proline, functioning of photosystem II as well as the changes in the antioxidative system was studied. Plants were subjected to either salt
(150, 300,450, 600 mM NaCl) or mannitol (300 mM) treatments for two weeks. Effects of iso-osmotic concentrations of NaCl (150 mM) and mannitol (300
512 mM) were compared in order to discriminate possible differences in C. ragusina response to ionic (NaCl) and osmotic (mannitol) components of salinity. C. ragusina plants, especially leaves, accumulated Na predominant osmotica – the concentrations of leaf Na and Cl -
+ and Cl as a
+
at 300mM NaCl treatment were 90- and 10-fold greater, respectively, than those measured in controls.
Simultaneously, significant reductions in plant K and Ca2 +
+ , Mg +
concentrations occurred. In addition, lower salt concentrations induced leaf succulence which was accompanied by increased leaf relative water status, vacuole volume and cell expansion. Regardless of significant accumulation of proline, 600mM NaCl- and mannitol-induced osmotic stress markedly decreased relative water content and caused plazmolysis in C. ragusina leaves. Given the suppressed levels of some photosynthetic proteins and down regulation of photosystem II efficiency, both stressors impaired photosynthesis though NaCl to a much lesser extent.
Activities of SOD, CAT and APX in salinized plants seem to play an essential protective role in the scavenging processes. In spite of induction of antioxidative defense, mannitol and higher salt treatments caused oxidative damage to lipids and proteins as evident by carbonyl content and lipoxygenase activity. Presented data confirm better adaptation of C. ragusina plants to salinity than drought, indicating its tolerance toward salinity through ion accumulation.
P0543 – ePoster
Study about biofertilizer application on physiological and biochemical performance of pigeon pea cultivars under salinity condition
1
Raouf Fard, F 1
F 3 , Behmanesh, M
, Omidbaigi, R 1 , Sharifi, M 2 , Sefidkon,
4
Dept of Horticulture, Faculty of Agriculture, Tarbiat
Modares University, Tehran, Iran; 2 Dept of Plant
Biology, Faculty of Biological Sciences, Tarbiat
Modares University, Tehran, Iran; 3 Research Institute of
Forests and Rangelands, Tehran, Iran; 4 Dept of
Genetics, Faculty of Biological Sciences, Tarbiat
Modares University, Iran
Anise hyssop ( Agastache foeniculum ) is a perennial, herbaceous medicinal and spice plant belonging to
Lamiaceae family. The herb is used for seasoning food and tea mixtures. The oil has been found to possess antimicrobial and anti-fungal properties, and it is also used in flavoring liqueurs and cosmetic industries. A. foeniculum , is known to contain significant amount of methyl chavicol. The effects of foliar application of different concentrations (10 –4 and 10 –3 M) of salicylic acid on essential oil compositon of Agastache foeniculum were studied. The experiment was conducted under the field condition at Zardband region in the north of Tehran. GC and GC-MS analyses revealed that the main identified components of essential oil were methyl chavicol,
Limonene, E-caryophyllene, Linalool, Octen-3-ol and germacrene D. Foliar application of salicylic acid with different concentrations resulted in quantitative differences in essential oil components. Salicylic acid treatments increased the methyl chavicol and limonene in comparison with untreated plants. The lower concentration (10 –4 M) was more effective on increasing

methyl chavichol amount than higher concentration (10 –3
M).
P0546 – ePoster
Chemical characterization of Brazilian propolis by
HPLC-MS and GC-MS: the black, red, and green
Brazilian varieties of propolis
Righi, A 1 , Negri, G 2 , Salatino, A 1
1 Botany Dept, Institute of Biosciences, University of São
2 Paulo, Brazil; Psycobiology Dept, Federal University of the State of São Paulo, Brazil
Propolis is a complex honeybee product with resinous aspect, containing plant exudates and beeswax. Their color, texture and chemical composition vary, depending on the location of the hives and exudates plant sources.
Relevant Brazilian propolis types currently recognized are the green and red propolis, the former presenting mainly prenylated phenylpropanoids and caffeoyl quinic acids and the latter isoflavonoids. The aim of present work was to provide chemical profiles of alcoholic and chloroformic extracts of nine samples of propolis, corresponding to seven regions of Brazil (Piauí, Goiás,
Alagoas, Bahia, Minas Gerais, São Paulo and Paraná).
Methanol and chloroform extracts were obtained and analyzed by HPLC/DAD/ESI/MS (high performance liquid chromatography with photodiode array detector coupled with electrospray ionization mass spectrometry) and GC/MS (gas chromatography coupled with mass spectrometry). The analysis of chromatographic profiles of all propolis extracts allowed the definition of three main types, already known, of Brazilian propolis directly related to their secondary metabolite classes: black
Brazilian propolis (BBP), from Piauí and Goiás states, rich in flavanones and glycosil flavones, red Brazilian propolis (RBP), from Alagoas state, containing isoflavonoids, pterocarpans, and chalcones, and green
Brazilian propolis (GBP), from Bahia, Minas Gerais, São
Paulo and Paraná states, showing prenilated phenylpropenoids as the major constituents as well as cafeoilquinics acids derivatives. Subsequently, the principal compounds of the black, red and green types were characterized by HPLC ESI/MS and GC/MS analysis, which are useful tools for qualitative analysis of marker compounds of Brazilian propolis. Besides that, these techniques were shown to be informatives tools for the analysis of crude propolis polar and non-polar extracts, respectively. Finally, it’s remarkable the prior identification of 2,4,3’,4’-tetrahydroxychalcone in RBP, flavones C-glycosides, such as apigenin-6-C-glucosyl-8-
C-arabinose (schaftoside) in GBP and triterpenes, such as
α
- and
β
-amyrinone and olean-12-en-3,11-dione in BBP.
P0547 – ePoster
Structure – activity relationship of indicine N-oxide and its functional derivatives by molecular orbital evaluation and charge flow dynamics
Robert, J 1 , Louis, C 2
1St St Joseph's College, Dept of Plant Biology and Plant
Biotechnology, Tiruchirappalli, Tamilnadu, India;
2 National Institute of Technology, Computational
Chemistry Lab., Tiruchirappalli, India
The antitumor activity of indicine N-oxide has been investigated for long due to the fact that it is free of some of the toxic effects associated with other pyrrolizidine alkaloids. The plant extract from Crotolaria biflora L.
(Twoflower Rattlebox) belonging to the family of
Fabaceae, showed the presence of indicine N-oxide. This plant extract showed a marked anti microbial activity too.
In order to correlate the structure–activity relationship, we report in this study, the evaluation of the molecular orbital energies of indicine N-oxide and its metabolites, along with the charge flow dynamics during its conversion. The important thermodynamic parameters for the antioxidant property such as excitation energy, dipole moment, oscillator strength have been computed by using time dependent density functional theory (TD-
DFT) at the B3LYP/6-31G(d) level. All computations have been carried out using Gaussian 03 package. The essential parameters for the antitumor behaviour such as ground state geometries, electronic transition energies and oxidation potentials are computed. A thorough analysis of the charge flow dynamics in the molecular orbital namely, highest occupied molecular orbital and lowest unoccupied molecular orbital, of these molecules has been carried out and presented to emphasize the role of these orbitals in effective charge separation, the important feature for the functionality of this molecule.
The charge flow dynamics of Indicine N-oxide and its analogues highlight the distinct correlation between the structure and the properties and hence the biological activity. The hybrid composition and occupancy of the
Natural Bond Orbitals (NBO) are analyzed to evaluate the changes in these parameters on biological changes.
NBOs are the localized 1 or 2 center orbitals. They effectively describe the molecular bonding pattern of electron pairs. The dipole moment of the molecules provide insight into the vastly varying characteristics of the molecules thought to be similar otherwise and hence the variation in their anti oxidant properties. Based on the results of our study it is reported that it is possible to mediate the antitumor activity of indicine N-oxide through structurally modified analogues, thereby totally avoiding the formation of the toxic intermediates, indicine and dehydroindicine. The role of the allylic ester and its analogues in the side chain is explored to bring about the required structural modification to reflect in the functional properties of the molecules. With its favorable ratio of desired activity to toxicity, indicine N-oxide stands apart from the other pyrrolizidine alkaloids and hence provides an excellent molecular framework for structural modifications. With a low excitation energy of
3.48 eV and a ëmax value of 356.4 nm, indicine N-oxide also promises as a candidate for photodynamic therapy.
Structural changes have been attempted to shift the ëmax to visible region characterized with a lower band gap.
P0549 – ePoster
Reduction of poisonous effects of nickel on soybean plants with application of ascorbic acid and gibberellin
Saeidi-Sar, S 1, 2 , Abbaspour, H 2
513

1 Dr Shariaty College, Holy Prophet Higher Education
Complex, Tehran, Iran; 2 Dept of Biology, Damghan
Branch, Islamic Azad University, Damghan, Iran
Effects of ascorbic acid (AsA) and gibberellins (GA3) protections were studied in soybean ( Glycine max L. cv.
Union Elf) plants under nickel (Ni) induced oxidative stress. The one-week old hydroponically-grown seedlings were exposed to NiCl
2
.6H
2
O (0, 0.25, 0.5 and 1.0 mM) either with or without AsA (1 mM) or GA3 (0.05 mM) or
AsA plus GA3, for 5 days. Total protein concentration increased in treatment of 0.25 mM Ni (with or without
AsA and/ or GA3), but in other treatments of Ni decreased in the absence of GA3, or AsA plus GA3.
Nickel induced oxidative damage in tissues, as evidenced by an increment in lipid peroxidation, with increasing lipoxygenase (LOX, EC 1.13.11.12) activity and malondialdehyde (MDA) production, and changes in activity of guaiacol peroxidase (GPX, EC 1.11.1.7). In the GA3- and/or AsA containing treatments, compensator effects were noticed. Native electrophoresis patterns of iso- guaiacol peroxidase (iso-GPX) in roots of
Ni- exposure plants showed that the thickness and staining intensity of iso-GPX were strongly increased in
GA3 plus AsA treatments respect to Ni alone. These results suggest that GA3 plus AsA treatment reduces Niinduced oxidative damage in soybean seedlings.
P0550 – ePoster
Gibberellic acid alleviates the cadmium toxicity in onion (
Allium cepa
L.) seedlings
Saeidi-Sar, S 1, 2 , Abbaspour, H 2
1 Dr Shariaty College, Holy Prophet Higher Education
Complex, Tehran, Iran; 2 Dept of Biology, Damghan
Branch, Islamic Azad University, Damghan, Iran
The effects of cadmium [Cd(NO
3
) 2 , 4H
2
O] and interaction of cadmium and gibberellic acid [GA3] on root and shoot growth, protein, chlorophyll contents and potassium concentration in onion ( Allium cepa L. cv.
Red Azarshahr) plant were studied. Seedlings were grown for 10 days in Hoagland nutrient solution containing variable Cd2 + concentrations (0, 0.1, 0.2, 0.4 mM) and GA3 (0, 0.03 mM). The ANOVA results showed that, with increasing cadmium concentration in nutrient solution significantly (P<0.01) root and shoot growth decreased. The adverse effect of Cd on growth was apparent from the reduction in dry weight and fresh weight in Cd-treated onion plants. As compared to the control, the decrease in dry weight and fresh weight was more pronounced in roots than the shoots. With application of gibberellic acid, these toxic effects of cadmium on growth reduced. Increased Cd concentrations in plants under Cd-stress were accompanied by declining concentration of potassium.
Addition of GA3 in the medium decreased of accumulation of Cd and increased K concentrations in the roots and shoots. The chlorophyll content and total protein content of shoot decreased in Cd–treated plants.
In the gibberellin-containing treatment compensator effects were noticed. This finding support that, application of gibberellic acid causes partial elimination of poisonous effects of cadmium on onion plants
514
P0551 – ePoster
Changes in the water status and osmotic solute contents in response to drought and salicylic acid treatments in four different cultivars of wheat
1
(
Triticum aestivum
L.)
Loutfy, N 1 , Sakuma, Y 1 , Inoue, M
Ehime University, Japan
1
Salicylic acid (SA) is an important signal molecule modulating growth and stress responses in plants. It also induces multiple stress tolerance in plants including that to drought. In this paper, four cultivars of wheat
( Triticum aestivum L.) were treated in three levels of drain (90% of maximum field capacity as control, and
60% or 30% as modestly or strongly drought-bearing treatments) to investigate the interactive effects of SA on their growth and various drought-stress parameters.
Under drought conditions, two tolerant cultivars Geza
164 and Sakha 69 maintained the greater dry-weight biomass especially in roots and leaf-relative water content (LRWC) than those of the sensitive Gemaza 1 and Gemaza 3. Drought stress generally decreased the biomass of root and shoot, LRWC, and content of inorganic solutes (Ca, K, and Mg) in all cultivars, while it caused a considerable increase in levels of carbohydrates, some proteins and organic solutes such as soluble sugars and proline. On the contrary, SA significantly increased the biomass of root and shoot,
LRWC values, and the total carbohydrate and protein contents in the plants, accompanying a conspicuous accumulation of the inorganic and organic solutes except proline. It was thus concluded in all cultivars that the SAstimulated osmotic regulation and dry matter biosynthesis occurred with increasing osmotic solutes but independently on proline biosynthesis. Moreover, these results signified the role of SA in regulating the drought response of wheat and as a potential growth regulator for improving the plant biomass under water stress. Droughtsensitive cultivars had the lowest levels of Ca and Mg, or
K especially in shoot. The possible roles of the ions and differences in four cultivars are also discussed.
P0552 – ePoster
Biosynthesis of polyhydroxybutyrate (PHB) in potato tuber
Sasaki, T 1, 2 , Ogasawara, M
, Shimada, H 3
1 , Yoshida, M 1 , Suzuki, H 1 ,
1
Matsumoto, K 3
3
Dept of Biological Science & Technology, Tokyo
University of Science, Chiba, Japan; 2 RNA Research
Center, Tokyo University of Science, Chiba, Japan;
Graduate School of Engineering, Hokkaido University,
Hokkaido, Japan
Plastics made from petroleum have highly processive and strength. However, since there are many problems about limitation of resource and CO
2
exhaust, petroleum independent technologies are necessary. Bioplastics are available for substitutional materials to petroleum plastics. Some kinds of bacteria species accumulate polyhydroxybutyrate (PHB) as an intracellular carbon storage. PHB can be used as bioplastics for use in a wide

range of agricultural, marine and medical applications because of its thermal plasticity and biodegradability.
Plants grow with only CO
2
, H
2
O and light, therefore they have economical advantages in the case of production of bioplastics satisfactory with carbon neutral. Until now, many research have been carried out to biosynthesize
PHB by introducing the PHB synthesis genes (
β
ketothiolase: PhaA, Acetoacetyle-CoA reductase: PhaB and PHB synthase: PhaC derived from Pseudomonas ) to plants, but failed in high accumulation of PHB. Many reports as well as our data show that the transformants harboring PHB synthesis genes driven by systemic promoter like 35S, resulted in apparent growth retardation. The function of sink organs like tuber or fruits are exclusively storage in massive amounts of sugars or starch, therefore, we considered that the productivity could be increased when PHB was produced at sink organs. Thus, potato tubers are applied for PHB accumulation because it functions as a sink organ specialized for starch storage. We established the efficient method to introduce transgenes to potato plants using stem of potato cultivar 'Sayaka'. We successfully obtained transformants that produce PHB. Patatin is a representative protein accumulated specifically in potato tubers. When the promoter of PhaC was changed to the patatin promoter from 35S promoter, PHB content of roots and tubers were increased. Then, we achieved
'tuber specific PHB production' with patatin promoter, although growth retardation was still observed. These results indicate that PHB production at nonphotosynthetic organs by using sink organ specific promoter is efficacious to increase of productivity.
P0553 – ePoster
Bio-efficacy of
Celosia argentea
L. and
Euphorbia hirta
L. leaf extracts on some pathogenic microorganisms
Saswade, R 1
1 Jijamata College of Science and Arts,
Dnyaneshearnagar, India
Celosia argentea and Euphorbia hirta are the two most abundant, invasive and fast growing weeds found during kharif and rabi seasons in semi-arid and arid agro climatic zones of Maharashtra State (India). Secondary metabolites produced by the weeds often influence the growth and development of certain microorganisms by inducing changes at cellular and molecular levels and also by influencing gene expression. Antimicrobial activities of these weeds also play a vital role in biological control of various pests and diseases. An attempt was made in the present investigation, to investigate the efficacy of two most abundant crop weeds, Celosia argentea and Euphorbia hirta , for their antimicrobial activity on some pathogenic microorganisms. Antimicrobial activity of Aqious
(0.02%), alcoholic (Ethanol and methanol, 0.01% in 90% ethanol) and dimethilformamide (0.01% in 90% DMF) leaf extracts of these two weeds were tested against the
Bacillus subtilis , Escherichia coli, Staphylococus aureus,
Candida albicans, Aspergillus fumigatus and Fusarium oxysporum , employing the method of Barry et al. (1986).
Experimental results revealed that the methanolic and dimethylformamide leaf extracts of Celosia were effective in inhibiting the growth of Bacillus , Fusarium,
Aspergillus and Candida . The ethanol and dimethylformamide extracts were effective in inhibiting the growth of Staphylococcus and Escherichia . All forms of Euphorbia leaf extracts inhibited the growth of
Bacillus , Staphylococcus and Escherichia . All the above extracts except the ethanol inhibited the growth of
Fusarium. The water, methanol and dimethylformamide extracts inhibited the growth in Aspergillus . Methanolic and dimethylformamide extracts inhibited the growth of
Candida . These results clearly establish the efficacy of these two weeds as efficient bio-controlling agents for checking the growth of certain pathogenic microorganisms.
P0554 – ePoster
Analysis of the regulatory mechanisms of woundinduced lignification in
Arabidopsis thaliana
Sato, Y 1 , Katsuno, F 2 , Ohmori, T 2
1 Biology and Environmental Science, Graduate School of
Science, Ehime University, Japan;
Faculty of Science, Ehime University
2 Dept of Biology,
Introduction – Lignin, a complex phenylpropanoid polymer, functions in development and defense in vascular plants. Lignification occurs in vascular tissues and in wounded or pathogen-infected sites. In this study, we proceeded with the analysis of the regulatory mechanisms of wound-induced (WI) lignification in
Arabidopsis thaliana .
Materials and Methods – For semi-quantitative analysis of lignification in wounded tissues, after wounding with scissors in cotyledons of Arabidopsis thaliana seedlings grown for 10 days on the germination medium agar, wounded organs were harvested in various periods, fixed with 90% ethanol, then stained lignin with phloroglucinol-HCl. Photomicrographs of lignified tissues were taken under constant conditions and analyzed the stained levels in wounded sites with image analysis software, Image J. As Arabidopsis strains, a wild type plant: Columbia (wt), ethylene insensitive mutants: ein4, etr1-1 and ein2-1, and a constitutive ethylene sensitive mutant: ctr1-1 were used. Furthermore, effects of an ethylene precursor: 1-aminocyclopropane-1carboxylic acid (ACC), an inhibitor of ethylene synthesis: aminooxyacetic asid (AOA), abscisic acid
(ABA), methyl jasmonate (MJ), various concentrations of NaCl or CaCl on WI lignin accumulation were
2 analyzed by addition of these substances in the medium.
Results – In wounded cotyledons of wt, WI lignin was deposited in the cut part of the veins. Wound-induced lignification was detected 1 day after wounding and the level was increased for 3 days. In ein4, increase in WI lignification was restrained 2 days after wounding in comparison to wt. In ein2-1, however, WI lignification was higher than wt. In ctr1-1, WI lignification was higher than wt. In wt, ACC increased WI lignification and AOA suppressed WI lignification. In ein4 and ctr1-1, ACC or
AOA did not affect WI lignification. In ein2-1, AOA or
ACC decreased WI lignification. In wt, ctr1-1, ein2-1 and ein4, MJ did not affect WI lignification. On the other hand, ABA increased WI lignification in wt, and extremely in ein4, ein2-1 and ctr1-1. In wt, 100–120 mM NaCl increased WI lignification, and 30–80 mM
515

CaCl
2
increased that in concentration-depended manner.
The increase in WI lignification with NaCl or CaCl also observed in etr1-1.
2
was
Discussion – These results suggest that ethylene is involved in the promotion of WI lignification. In this study, relationship between WI lignification and MJ was not observed. On the other hand, ABA was also shown to promote WI lignification. For further analysis, ABA insensitive mutant: abi1 will be used. EIN2 is thought to function as the junction of ethylene and ABA signal pathway. Therefore, increase in WI lignification in ein2-
1 may concern endogenous ABA. Recently, involvement of map kinase cascade on ethylene signal pathway was suggested. Mutants of the map kinase kinase will be used for further analysis.
P0555 – ePoster
Contrasting strategies of aluminium accumulation in mistletoes growing on aluminium-accumulating and non-accumulating hosts in the savannas of central
Brazil
Scalon, MC 1 , Haridasan, M 2 , Franco, AC
1 Macquarie University, Australia; 2
2
Universidade de
Brasília, Brazil
In savannas of Brazil, Al-accumulation is a common strategy in some plant families. Mistletoes offer a unique model to study the effects of Al on plants, because several species are able to grow and reproduce on both
Al-accumulating and non-accumulating hosts. We investigated the patterns of Al and nutrient uptake in two mistletoes, Psittacanthus robustus specific to Alaccumulator hosts and Phthirusa ovata which parasitizes
Al-accumulating and non-accumulating hosts. We measured foliar concentrations of N, P, K, Ca, Mg, Cu,
Fe, Mn, Zn and Al in leaves, seeds and branches of
Phthirusa ovata and Psittacanthus robustus infecting
Miconia albicans, an Al-accumulating species, and P. ovata infecting Byrsonima verbascifolia , a non-Alaccumulating species. We also harvested leaves of the mistletoes for histological analysis using hematoxylin dye for localization of Al in tissues. Passive nutrient uptake in mistletoes was evidenced by the significant correlation between parasite and host in leaf concentrations of Ca, Mg, Mn, Cu and Al, but not of N, P and K. Parasitic plants had higher leaf K than hosts, whereas leaf N and P were correlated in hosts, but not in hemiparasites. Although passive nutrient and Al uptake occurred in both hemiparasites, they exhibited contrasting behaviours in relation to Al compartmentalization. Al was found in large concentrations in leaves, seeds and branches of P. robustus . In P. ovata , despite the high Al concentration in the leaves when attached to the Al-accumulating host, there was no accumulation in seeds or branches. Leaf anatomic analysis indicated that Al was stored in the epidermic cells in P. ovata , while Al was present in many different structures in P. robustus . This element is probably immobilized in the leaf tissue in P. ovata , while it is retranslocated to different plant organs in P. robustus , suggesting that the host specificity of this species might be related to large amount of aluminium presence in the host tissues.
516
P0556 – ePoster
Host phenological behavior affects water relations of a mistletoe (
Phthirusa ovata
(Pohl.) Eichler –
Loranthaceae) in the savannas of central Brazil
Scalon, MC 1 , Rossatto, DR 2 , Franco, AC 2
1 Macquarie University, Australia;
Brasília, Brazil
2 Universidade de
Several mistletoes species are able to grow and reproduce on both deciduous and evergreen hosts, which suggest a certain degree of plasticity to coupe with intrinsic functional characteristic of their hosts. The aim of this study was to understand the different strategies of a mistletoe species growing on deciduous and evergreen hosts. The study was conducted in a savanna field site at the IBGE Ecological Reserve (15° 56' 41' S and 47° 53'
07'W), where we sampled 5 individuals of Phthirusa ovata infecting Miconia albicans , an evergreen species, and 5 individuals infecting Byrsonima verbascifolia , a deciduous species. We measured diurnal cycles of gas exchange, pre-dawn and midday leaf water potential, water use efficiency and stomatal anatomical traits. The measurements were made during the dry and the wet season. Transpiration rates during the dry season were higher in P. ovata attached to B. verbascifolia when compared to individuals on M. albicans . The opposite situation occurred in the wet season, with higher transpirations rates measured in P. ovata growing on the evergreens hosts. In general, the hosts presented higher water use efficiency compared to the mistletoes in both seasons. Leaf water potential values were similar between the hemiparasites and the hosts in the rainy season, but mistletoes showed lower values during the dry season, without differences when attached to evergreens or deciduous hosts. Despite differences in daily stomatal conductance values, with the mistletoes showing higher values, there was no difference among parasites and its hosts in rates of maximum CO
2 assimilation (Amax), and stomatal conductance at Amax.
There was a tendency of related responses in stomatal closure within hosts and their respective parasites, which suggest a conservative use of water of mistletoes in habitats with drought conditions, such Cerrado. P. ovata presented stomata on both sides of the leaf surface, with higher stomata density in the abaxial surface. Mistletoes attached to the deciduous host showed significantly lower stomatal density. Therefore, we found differences in the behavior of P. ovata (e.g. transpiration rates and stomatal density) when attached to different phenological group host. This can show some phenotypical plasticity
(anatomical and physiological), that could allow this parasitic plant to grow on a large variety of hosts.
P0557 – ePoster
Comparative analysis of transcriptomic responses to
1 nutrient starvation between mono- and di-cotyledons
Secco, D 1 , Narsai, R 2 , Wang, C 2 , Shou, H 3 , Whelan, J 1
ARC Centre of Excellence in Plant Energy Biology,
University of Western Australia; 2 State Key Laboratory of Plant Physiology and Biochemistry, College of Life
Sciences, Zhejiang University, China; 3 Joint Research

Laboratory In Genomics and Nutriomics, College of Life,
Zhejiang University, China
Plant growth and development is highly influenced by macro- and micro-elements available in the soil solution.
During their life cycle, plants often face depletion of one or more essential element, limiting their growth.
Therefore, understanding the complex responses of plants to nutrient starvation is a critical step to improve their yield. So far, research has mainly been focusing on individual nutritive elements, instead of looking at a more global view. In order to untangle the complex cross-talk between individual nutritive elements, a comparative approach was undertaken using Arabidopsis as a model. Publicly available microarrays under specific nutrient deficiencies were compared, with particular focus on responses to iron and phosphate deficiency.
These analyses identified overlapping responses to nutrient deficiency and revealed subsets of genes specifically responsive to individual nutrients. Moreover, in order to compare if mono- and di-cotyledons shared similar responses to nutrient starvations, this transcriptomic analysis was also performed in rice.
Preliminary results revealed common and distinct sets of genes responsive to nutrient deficiency, indicating that monocots and dicots may have evolved different strategies to cope with low nutrient availability.
P0558 – ePoster
Structure activity relationship study of strigolactones as a shoot branching-inhibiting hormone in rice
Umehara, M 1
Yamaguchi, S 1
, Akiyama, K 2
1 RIKEN PSC, Japan; 2
, Seto, Y 1 , Hanada, A 1 ,
Osaka Prefecture University,
Japan
Strigolactones (SLs) were first discovered as seed germination stimulants of root parasitic plants such as
Striga and Orobanche . Later, SLs were identified as plant-derived signals in the symbiosis with arbuscular mycorhizal fungi that supply inorganic nutrients to the host plant. More recently, SLs or their metabolites have been identified as a novel plant hormone that inhibits shoot branching. However, little is known about the structural requirements of SLs for shoot branching inhibition. To address this question, we performed a structure-activity relationship study of various SL analogs in shoot branching assays using a SL-deficient rice mutant. SLs have a four-ring structure, with a tricyclic lactone (ABC-rings) being connected to a methylbutenolide moiety (D-ring) via an enol ether bridge. Among four stereoisomers of 5-deoxystrigol
(5DS), (+)-5DS and (-)-2'-epi-5DS, whose C-2' configuration is R, showed higher biological activity compared with the 2'S-isomers. Likewise, (+)-GR24 and
(-)-2'-epi-GR24 showed stronger activity among the four stereoisomers of GR24. Therefore, the R configuration at
C-2' is crucial for shoot branch inhibition. We found that
3, 6'-dihydro GR24, a GR24 derivative whose double bond of the enol ether bridge is reduced to a single bond, was not active. A reduction of the 3',4'-double bond in the D ring to a single bond also resulted in a severe decease in biological activity. By contrast, a truncation of the AB-rings significantly increased, rather than decreased, shoot branching inhibition activity. Our results show that the AB part of SLs is dispensable, while the enol ether linked to ring D is critical for biological activity in rice.
P0559 – ePoster
A Novel Factor rice FLOURY ENDOSPERM 2 is involved in regulation of gene expression for storage substances
She, KC 1,2 , Kusano, H 1
Yamakawa, H 3 , Hakata, M
, Shimada, H 1,2
, Tsurumaki, Y
3
1 , Yaeshima, M 1
, Aoyama, T 4 , Sasaki, T 1,2
1
Satoh, H 5
3
Dept of Biological Science & Technology,Tokyo
University of Science, Chiba, Japan; 2 RNA Research
Center, Tokyo University of Science, Chiba, Japan;
,
,
4
National Agricultural Research Center, Nigata, Japan;
Institute for Chemical Research, Kyoto University,
Kyoto, Japan; 5 Faculty of Agriculture, Kyushu
University, Hukuoka, Japan
Rice endosperm accumulates a massive amount of storage starch and storage proteins during seed development. However, little is known about the regulatory system involved in the production of storage substances. Mutants showing abnormal features of storage starch in the endosperm, such as floury, whitecore and opaque kernel phenotypes have been isolated.
The flo2 grains showed white and floury features with smaller grain size compared with wild type. Scanning electron microscopy (SEM) images of transverse sections of flo2 grains indicated that the endosperm of the flo2 mutant was filled with loosely packed starch granules composed of small and spherical-shaped granules with large air spaces, while the wild-type endosperm consisted of densely packed starch granules composed of large and irregular polyhedral-shaped granules. In the endosperm of the flo2 mutant, amylose content was lower than the wild type, amylopectin chain length distribution profile also altered. The flo2 mutant exhibits extremely reduced levels of BEI expression in developing rice endosperm along with decreased levels of other starch-synthesizing enzymes including AGPase, GBSS, SS and BEIIb. The flo2 gene is suggested to be a transcriptional regulationfactor widely involved in endosperm development. We mapped the flo2 locus using the F2 progenies crossed between a flo2 mutant EM37, which has been made by treatment with MNU on a japonica cultivar kinmaze, and an Indica cultivar Kasalath. Our map-based cloning identified FLOURY ENDOSPERM 2 (FLO2), a member of a novel gene family, as the gene responsible for the rice flo2 mutation, which was evolutionally conserved in plants. Introduction of the FLO2 gene into the flo2 mutant restored the small grain size and aberrant shape.
SEM images of the transverse section of the grains of the transformants indicate rugged surfaces as observed in those of the wild-type endosperm, which consisted of densely packed polyhedral-shaped starch granules.
Amylose content and amylopectin chain length distribution profile was mostly complemented in transformants. These transformants produced similar amount of glutelins, BEI, and RA16 in the endosperm, as observed in those of the wild-type plant. These results indicate that the loss-of-function mutation of FLO2 gene invokes the phenotype of the flo2 mutation. FLO2 was
517

abundantly expressed in developing seeds coincident with production of storage starch and protein, as well as in leaves, while abundant expression of its homologs was observed only in leaves. The flo2 mutation decreased expression of genes involved in production of storage starch and storage proteins in the endosperm. Yeast-2hybrid demonstrated the FLO2 interact with a bHLH protein, a predicted transcription factor. Over-expression of FLO2 enlarged the size of grains. These results suggest that FLO2 plays a pivotal regulatory role in expression for storage substances and affecting rice grain size and starch quality.
P0560 – ePoster
CAS is a chloroplast protein regulating biosynthesis of salicylic acid and plant immune signaling
Nomura, H
Fukusaki, E
3
2
, Uemura, S 1
S 1 , Takebayashi, K 2
, Shiina, T 1
, Kanda, Y 1 , Nakahira, Y 1
, Sugimoto, T 2
, Sano,
, Yoshioka, H 3 ,
1 Kyoto Prefectural University, Kyoto, Japan;
University, Osaka, Japan;
Japan
3
2 Osaka
Nagoya University, Nagoya,
There has been a much debate as to whether chloroplasts are involved in plant immune response. Here we show that microbe-associated molecular pattern (PAMP) signals are quickly relayed to chloroplasts and evoke a transient Ca2 + oscillation in stroma. The stromal Ca2 oscillations are dependent on a thylakoid associated Ca2
+
+ binding protein CAS. Furthermore, CAS is required for induction of plant basal resistance, referred as PAMPtriggered immunity (PTI), such as stomatal closure, callose deposition and PR1 gene expression. Chloroplastlocalized CAS also regulates effector-triggerd immunity
(ETI), which results in rapid local cell death (hyper sensitive responses; HR). Salicylic acid (SA) is a key signal molecule in regulating plant defense against pathogens. SA is synthesized in chloroplasts via chloroplast – localized isochorismate synthase 1 (ICS1) in Arabidopsis . Further characterization of CAS knockout Arabidopsis revealed that CAS is indispensable for PAMP (flg22)-induced biosynthesis of salicylic acid
(SA), suggesting that CAS is a novel chloroplast factor regulating SA biosynthesis. In order to elucidate the role of CAS in regulating SA biosynthesis, we examined expression of genes involved in the regulation of SA accumulation. We found that some genes essential for
SA biosynthesis including ICS1, PAD4 and EDS5 was impaired in CAS KO mutants. These results suggest that chloroplasts provide control over the expression of nuclear-encoded SA-related genes and CAS plays a critical role in this novel retrograde signaling. This study reveals the profound role of chloroplasts in mediating plant immune signaling.
P0561 – ePoster
Arabidopsis
Ital Qa-SNARE SYP2 proteins localized to different subcellular regions function redundantly in vacuolar protein sorting and plant development
Shirakawa, M 1 , Ueda, H 1 , Shimada, T 1
Shimada, TL 1 , Kondo, M 2
Nishimura, M, 2
, Takahashi, T
, Hara-Nishimura, I 1
3
, Koumoto, Y 1 ,
, Okuyama, Y 4 ,
518
1 Kyoto University, Japan;
Biology, Japan; 3
2 National Institute For Basic
Okayama University, Japan;
Museum of Nature and Science, Japan
4 National
SYP2 proteins are a subfamily of Qa-SNAREs (soluble
N -ethylmaleimide sensitive factor attachment protein receptors) that are possibly responsible for protein trafficking between prevacuolar compartments (PVC) and vacuoles.
Arabidopsis thaliana SYP22/VAM3/SGR3 and SYP21/PEP12 proteins function independently, and are yet both reported to be essential for male gametophytic viability. Here, we systematically examined the redundancy of three SYP2 paralogues (i.e.,
SYP21, 22, and 23) using a Col-0 ecotype that harbored a SYP2 paralogue (SYP23/PLP) that lacked a transmembrane domain. Surprisingly, no visible phenotypes were observed, even in the double knockout syp21/pep12 syp23/plp . A deficiency of either
SYP21/PEP12 or SYP23/PLP in a syp22 background resulted in a defect in vacuolar protein sorting, which was characterized by an abnormal accumulation of protein precursors in seeds. SYP21/PEP12 knockdown enhanced the syp22 phenotype (i.e., semi-dwarfism, poor leaf vein development, and abnormal development of myrosin cells), and the additional knockout of
SYP23/PLP further aggravated the phenotype. A GFP-
SYP23/PLP fusion localized to the cytosol, but not to the
PVC or vacuolar membrane, where SYP21/PEP12 or
SYP22/VAM3 localized, respectively. The immunoprecipitation analysis showed that SYP23/PLP interacted with vacuolar Qb- and Qc-SNAREs, VTI11 and SYP5, respectively, suggesting that SYP23/PLP is able to form a SNARE complex anchoring the membrane. Unexpectedly, we found that the multicopy expression of a genomic fragment of SYP23/PLP suppressed the abnormal syp22-3 phenotype. Thus, SYP2 proteins, including cytosolic SYP23/PLP, appear to function redundantly in vacuolar trafficking and plant development. (1) Shirakawa, M. et al. (2009). (2)
Shirakawa, M. et al. (2010).
P0563 – ePoster
Growth, antioxidant defence systems, lipid peroxidation and proline content in soybean genotypes under salt stress
Siddiqi, TO 1 , Khan, F 1 , Ahmad, A 1 , Mahmooduzzafar 1
1 Dept of Botany, Faculty of Science, Hamdard
University, New Delhi, India
The present investigation was conducted for evaluating genotypic variation in nine genotypes of soybean
( Glycine max L.) under salt stress. Ten-day-old seedlings, grown hydroponically, were treated with 7 levels of NaCl
(0, 25, 50, 75, 100, 125 and 150 mM). After 5 days of treatments, growth, lipid peroxidation and enzyme activities of antioxidant defence system were studied.
Growth, measured in terms of length, fresh weight and dry weight of plants, was drastically reduced in PK-416, while there was little effect of NaCl treatment on Pusa-37 genotype of soybean. High level of lipid peroxidation was observed in PK-416 as indicated by increased level of malondialdehyde. Activities of superoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase were maximum in Pusa-37 where 9-fold, 1-fold, 5-fold

and 6-fold increase over control were observed, respectively. It can be revealed that PK-416 and Pusa-37 genotypes of soybean are salt-sensitive and salt-tolerant, respectively, and antioxidant defence system involved in conferring the sensitiveness and tolerance in these genotypes.
P0565 – ePoster
Alterations in endogenous pools of diterpenes cafestol and kahweol in in vitro cultures of
Coffea canephora under the influence of inhibitors of some plant growth regulators
Sridevi, V 1 , Giridhar, P 1 , Ravishankar, GA 1
1 Central Food Technological Research Institute (CSIR,
India), Mysore, India
Cafestol and kahweol are the two unique diterpenes associated with the unsaponified lipid fraction of coffee brew and are reported to be responsible for an increase in serum cholesterol and triglycerides which are known to increase the risk of coronary diseases and also induce damage to hepatocytes in people who consume more coffee. The content of cafestol and kahweol in coffee is significantly influenced by agronomical, environmental factors and its level in coffee drink depend on coffee bean processing and brewing. It is well established that growth regulators, ethylene and polyamine inhibitors influence endogenous pools of caffeine in in vitro cultures of Coffea sp. There are no clear information on the actual biosynthetic pathway of the cafestol and kahweol. However, it is predicted that their biosynthesis is from a precursor of geranylgeranyldiphosphate
(GGDP). A study was taken up to find out the variations in endogenous pools of cafestol and kahweol in somatic embryos of robusta coffee ( Coffea canephora CxR variety) under the influence of some antagonists of plant growth regulators such as antiauxins (2,3,5triiodobenzoic acid -TIBA), anticytokinin (lovastatin), antigibberellin (paclobutrazol), ethylene inhibitors (silver nitrate, cobalt chloride). Respective inhibitors of plant growth were supplemented to the Murashige and Skoog medium at different concentrations to culture somatic embryos. Upon 8 weeks of culture, somatic embryos were harvested for analyzing their cafestol and kahweol profiles through HPLC, which revealed a significant reduction in both diterpene especially under the treatment of TIBA, lovastatin and silver nitrate. There was 60-75% reduction in both cafestol and kahweol in presence of TIBA, followed by 56-62% reduction in presence of silver nitrate. But there was 25-32% elevation of cafestol and kahweol in presence of paclobutrazol. This may be due to the gibberellin inhibitory nature of paclobutrazol that could possibly rise entkauronic acid pool which is also a precursor for cafestol and kahweol. In order to elucidate the actual role of these inhibitors of plant growth on alterations in diterpenes pools of somatic embryos, in depth investigations at physiological levels are vital and the leads obtained in this direction will be presented.
P0566 – ePoster
Role of C-terminal fragment in membrane-bound
β
glucosidase from
Physarum polycephalum
Tada, S 1
Y 1
, Moriyasu, M 1 , Hayase, M 1 , Hase, T 2 , Minami,
1 Okayama University of Science, Okayama, Japan;
2 Institute for Protein Research, Osaka University, Osaka,
Japan
Physarum polycephalum , one of the true slime molds, inhabits under rotten trees or dead leaves in the forests.
Physarum has a unique life cycle that includes a diploid plasmodium. The plasmodia express three forms of
β
glucosidases: intracellular soluble, membrane-bound, and secretory enzymes. We particularly have focused on the membrane-bound
β
-glucosidase, BglM1, of molecular mass 130 kDa. The primary structure of BglM1 consists mainly of two regions: the N-terminal region that contains typical glycosyl hydrolase family 3 domains and the C-terminal region that has a unique sequence unrelated to hydrolytic reactions. Notably, the C-terminal region of BglM1 contains two calx-
β
motifs known as calcium binding sites, an RGD sequence, and a transmembrane region. Thus, Physarum BglM1 is a new type of
β
-glucosidase that differs from all previously identified family 3 enzymes. BglM1 is only present in the plasmodia and microplasmodia during the life cycle.
Observations using immunofluorescence microscopy indicate that BglM1 locates on the cell membrane. The molecular mass of native BglM1 in microplasmodia is estimated to be 230 kDa, although 130 kDa is the value calculated from the amino acid sequence. During BglM1 purification, its apparent size gradually decreases from
230 to 130 kDa. However, when BglM1 is purified in the presence of calcium ions, the molecular mass of the enzymes is maintained at 230 kDa. Furthermore, we also purified the Physarum secretory
β
-glucosidase that has similar properties to BglM1 and cross-reacts with two antibodies specific for the N-terminus or C-terminus of
BglM1. Interestingly, as in the case of BglM1 purification, the molecular mass of secretory enzyme purified in the presence of calcium ions is approximately
200 kDa and in the absence of calcium ions is approximately 130 kDa. After purification, the 130-kDa molecule gradually degrades to 95 kDa. Because the 95kDa molecule does not react with the C-terminal antibody of BglM1, it is probably missing a part of the
C-terminal region. Further, the 130 kDa molecule can bind to microcrystal cellulose (Avicel, SIGMA Life
Science) in the absence of calcium ion, but not in the presence of calcium ions. However, the 95-kDa molecule never binds to cellulose. From these results, we suggest that calcium ions influence the conformation of the whole molecule through the C-terminal region of BglM1.
Currently, we are attempting to analyze the structural differences between the 95-kDa and 130-kDa molecule.
To examine the function of the C-terminal region in
BglM1, some recombinant BglM1s were expressed in E. coli . We aim to analyze the properties and
β
-glucosidase activity of these recombinant proteins. Based on these results, we will report on the role of the C-terminal region in BglM1.
P0567 – ePoster
The effect of two plant growth regulators on callus generation of
Oryza sativa
L. var. Hashemi
Tajadod, G 1 , Farzamei, M 2 , Kalamei, Z 1
519

1 Islamiz Azad University, North Tehran Branch, Iran;
2 Islamic Azad university, Saveh Branch, Iran
Rice ( Oryza sativa L .) is the world most important cereal crop after wheat and maize. Rice has 24 species.
Oryza sativa ( Oryza sativa L., 2n=2x=24) is an annual grass. It belongs the family Gramineae. In this research, seed of rice ( Oryza sativa L. var. Hashemi) were inoculated on to MS medium containing various concentration of 2.4-D:)0,1,1.5,2,3mg/l (and
Kn:)0,0.5,1,1.5mg/l (dfferent part of 7 days old seedling were grown on MS medium that supplemented by different concentrations of 2 hormons. Different explants that used for callus regenerataion were root,stem,hypocotyle and epicotyle . Our results showed that explants of rice in hormonal concentration:2,4-
D(2mg/l)+Kin (1mg/l) and 2,4-D(2mg/l)+Kin (1.5mg/l) and 2,4-D 3mg/l formed more and biggest callus.
Between different explants that used, epicotyle
,hypocotyle and root had more potential for callus regeneration. In this research use cross section of Oryza sativa .
P0568 – ePoster
The effect of four growth regulators on Callogenesis and Organogenesis of
Daucus carota
L. var
α
Tajadod, G 1 , Majd, A 1 , Salimpour, F 1 , Sattaripour, M 1
1 Islamiz Azad University, North Tehran Branch, Iran
Carrot belongs to the Apiaceae family and is called
Daucus carota L . It is a biennial plant with erect roots and hair-free stems. Seeds of Daucus carota L. var. á were cultivated in MS medium. Hypocotyles were chosen as explants in three different developmental stages ( seven, twelve, and seventeen days old). Plantlets which were induced from these explants were placed in
MS medium, supplemented by different hormonal concentration. Four hormones were used ( 2,4,D : 0–1–2 and 3 mg/l, kin : ( 0–0.5 and 2 mg/l ), NAA: ( 0–0.5–1 and 2 mg/l) and BAP : ( 0–0.5–2 and 4 mg/l).
Hypocotyles treated with , kin 2 mg/l, BAP 4 mg/l, NAA
1 mg/l, and 2,4,D 2 mg /l formed more and bigger calli.
2 and 7 days old root pole hypocotyles produced more calli, compared to stem pole ones. In media with 2 mg/l kin, after appearance of calli organogenesis was observed too, while this was not been observed in other media.
P0569 – ePoster
Tolerance to flooding and salinity in
Melilotus siculus is associated with highly porous phellem and leaf gas films
Teakle, N 1 , Pedersen, O 1 , Verboven, P 1 , Colmer, T 1
1 University of Western Australia, Australia
Aerenchymatous phellem (secondary aerenchyma) has rarely been studied in roots and its role in tolerance to flooding is not clear. Melilotus siculus , an annual legume of wet saline land, develops phellem on roots and hypocotyl that is critical for internal aeration. This study examined the 3D structure of phellem and the response of M. siculus to combined submergence and flooding. X-
520 ray computed tomography (CT) was used to visualise the
3D microstructure of M. siculus roots and to quantify O diffusion through phellem and stele tissue. Radial O
2
2 profiles through roots and respiration of separated stele and phellem tissue were measured using O
2 microelectrodes. Responses to submergence, at a range of salinity levels, were also studied with leaf gas films removed or intact. The porosity of phellem was calculated from both X-ray CT and the buoyancy method to be over 50% for stagnant-treated M. siculus, due to the presence of large gas spaces (aerenchyma). The radial O highest in the phellem, approaching that of O
2
10 -5 m2 s -1 ). Radial O
2
2 diffusivity calculated from the micro-CT images was
in air (1.6
profiles also revealed high O
2
in the phellem, with the stele severely hypoxic (<0.2 kPa).
M. siculus survived up to one week of complete submergence combined with salinity (up to 100 mM
NaCl). Removal of gas films reduced underwater photosynthesis and increased leaf senescence. These results highlight the importance of aerenchymatous phellem and leaf gas films for tolerance to flooding and salinity in M. siculus.
P0571 – ePoster
Physiological responses of Iranian alfalfa ecotypes to
3
1 salt stress
Torabi, M 1 , Halim, R 2 and Natural, Iran; 2
, Chokan, R 3
Isfahan Center For Research of Agricultural Science
University Putra Malaysia;
Seed and Plant Improvement Institute of Iran
In order to assess physiological responses of Iranian alfalfa ecotypes to salt stress a hydroponic greenhouse trial was conducted in University Putra Malaysia. The seeds of five ecotypes were planted in polybags filled with sand and immersed in nutrient solution. The EC of nutrient solution was increased gradually by adding sodium chloride. The EC at each stage of salt addition was 6, 12, and 18 ds m -1 and the physiological characteristics of alfalfa ecotypes were measured at every stage of salt. The results showed that the physiological parameters were affected by salt concentration and there were varying responses between genotypes. Salt stress had a negative impact on crop growth rate in all ecotypes. Salt stress resulted in a linear decline in the photosynthetic rate from 21ìmol/m2/s in control to 5
µ mol/m2/s at 18 ds m -1 . A positive correlation between photosynthetic rate and stomatal conductance was observed. The chlorophyll content decreased with increasing salinity but there was no a significant difference between ecotypes in terms of chlorophyll content. The results of Fv/Fm, ratio showed that up to
12 ds m-1 there was no significant change in Fv/Fm ratio but after 12 ds/s the Fv/Fm ratio decreased with increasing of salt stress. Overall the study indicated that ecotypes showed difference in their physiological response to salt stress and ecotype number 2 and 5 were superior between five ecotypes.
P0572 – ePoster
Contribution of the methylerythritol phosphate pathway to strigolactone biosynthesis

Umehara, M 1
Hishiyama, S 4
, Xie, X 2
, Yokota, T 3
Yamaguchi, S 1
, Uchida, K 3 , Hanada, A 1
, Yoneyama, K 2 , Kasahara, H 1
,
,
1 RIKEN Plant Science Center, Japan;
Center, Utsunomiya University, Japan;
Biosciences, Teikyo University, Japan;
2 Weed Science
3
4
Dept of
Forestry and
Forest Products Research Institute, Japan
Strigolactones (SLs) are a group of terpenoid lactones, and function as a plant hormone that inhibits shoot branching as well as communication signals for symbiosis and parasitism in the rhizosphere. SLs consist of tricyclic-lactone (ABC-ring) and methyl butenolide
(D-ring), connected with an enol ether bridge. Studies using chemical inhibitors and mutants have suggested that the ABC-ring is derived from a carotenoid cleaved product. However, the isoprenoid origin of the SL carbon skeleton has not been proved directly in a metabolism study. Especially, little is known about the biosynthetic origin of D-ring. To address this question, a 13C-labeled precursor specific to the mevalonate or methylerythritol phosphate (MEP) pathway was fed to Lotus japonicus cultured roots in the presence of a chemical inhibitor that blocks each endogenous pathway. LC-MS/MS and 13C-
NMR analysis of 5-deoxystrigol, a previously identified
SL in this species, indicated that the MEP pathway contributes to the biosynthesis of the entire SL carbon skeleton, including the D-ring. Possible pathways for SL biosynthesis will be discussed.
P0575 – ePoster
Immunohistochemical analysis of the localization of an auxin efflux facilitator, CsPIN1, during gravimorphogenesis of cucumber seedlings
1
Watanabe, C 1 , Fujii, N 1
Koshiba, T 2
Japan; 2
, Nishimura, T 2 , Miyazawa, Y 1
, Takahashi, H 1
,
Graduate School of Life Sciences, Tohoku University,
Dept of Biological Sciences, Tokyo Metropolitan
University, Japan
When cucumber seeds are placed in a horizontal position for germination, resulting seedlings develops a specialized protuberance, termed the peg, on the lower side of the transition zone between the hypocotyl and the root, due to gravistimulation. The peg anchors the seed coat so that the elongation of hypocotyl pulls the cotyledons out of the seed coat. On the other hand, when cucumber seeds germinate in a vertical position with the radicle pointing down or under microgravity conditions in space, the seedlings develop a peg on each side of the transition zone. Thus, cucumber seedlings have a potential ability to develop a peg on each side, and peg formation on the upper side of the transition zone is suppressed in response to gravity when seedlings are grown in a horizontal position on the ground. We previously showed that application of either auxin or auxin efflux inhibitors induced peg formation on both sides of the transition zone even when the seedlings were placed horizontally. Thus, we hypothesized that gravistimulation induces auxin efflux to decrease auxin content on the upper side so as to suppress peg formation. However, the molecular mechanism underlying this process is less understood. To reveal the mechanism of how gravistimulation induced this asymmetric auxin redistribution in the transition zone, we analyzed the localization of CsPIN1, an auxin efflux facilitator that was expressed in the endodermal cells, which were thought to sense gravistimulation. The transition zone has four vascular strands, and endodermis situates just outside of each vascular strand. In the seedlings grown in a vertical position, a peg developed on each side of the transition zone, and CsPIN1 signals in endodermal cells among the four endodermal layers were equally detected on the vascular bundle side. In the seedlings grown in a horizontal position, CsPIN1 signals in the two endodermal layers in the upper side of the transition zone were much stronger than those in the lower side. In addition, when the vertically grown seedlings were reoriented and placed horizontally, this differential accumulation of CsPIN1 protein commenced within 30 min after gravistimulation. Furthermore, our measurement of endogenous auxin content revealed that asymmetric auxin distribution was induced within 30 min after gravistimulation. Our results suggest that the gravity-induced changes of CsPIN1 localization in endodermal layers participate in regulating auxin distribution that leads to unilateral positioning of a peg in cucumber seedlings.
P0577 – ePoster
1
Phylogenetic relationships of the Korean
Salix
species
Won, H 1 , Jin, BB 1 , Lim, CK 1
Daegu University, Republic of Korea
We have analyzed chloroplast and nuclear DNA marker sequences of Korean Salix species to reveal the phylogenetic relationships among them. Plastid trn L intron, trn L-F IGS, trn Hpsb A IGS, atp Fatp H IGS , psb Kpsb I IGS, rpo B gene, rpo C gene, and mat K sequences are tested and analyzed, in addition to nuclear xdh gene sequences. Plastid mat K and nuclear xdh produced best resolution and most usable. Using both the sequences, we have reconstructed the phylogenetic relationships among the Korean Salix species, including other relevant species.The resulting phylogeny matches with previous phylogenetic studies and indicates a evolutionary pathway of an Korean edemic species.
P0578 – ePoster
Water supply and photosynthesis in terrestrial, climbing and semi-epiphytic
Freycinetia excelsa
F.
Muell (Pandanaceae)
Yansen, Y 1, 2 , Congdon, RA 1 , Holtum, JAM 2
1 School of Marine and Tropical Biology, James Cook
University, Australia; 2 Dept of Forestry, University of
Bengkulu, Indonesia
Freycinetia excelsa is a secondary hemi-epiphytic vine with a three phase life-cycle: terrestrial, terrestrial but climbing a host, and semi-epiphytic. Beginning life as ground-dwelling plant with normal roots, F. excelsa climbs a host and the basal portion of the stem dies back, in most cases remaining connected to the soil via aerial feeder roots. We ask whether water supply and
521

photosynthesis differs between the three morphological forms of F. excelsa . For all growth forms of F. excelsa , pre-dawn leaf water potentials correlated with volumetric soil moisture, and leaf water potential and CO
2 assimilation rates were generally lower in the dry season than in the wet season. Within each season, water potentials of leaves from all growth forms were similar but the patterns of daily CO
2
CO
2
exchange differed, with
uptake by terrestrial plants most affected by dry season conditions. Terrestrial plants exhibited the lowest
CO
2
exchange in the dry season, but the CO
2
exchange rates were similar for the three forms during the wet season. Stem hydraulic conductivity and xylem anatomy differed between the growth forms. Terrestrial plants, with conventional roots and a main stem, have narrow xylem and lower hydraulic conductivity. In comparison, climbing and semi-epiphytic plants are attached to the soil by narrow aerial roots and hydraulic conductivity of the stem is greater. Down-regulation of CO
2
exchange in the dry season was greatest in the terrestrial plants, but all forms operated at similar pre-dawn water potentials of
-1.5 MPa and afternoon water potentials of not less than -
2 MPa. The lower hydraulic conductivity of the stems of terrestrial plants may restrict water supply such that down-regulation of CO
2
uptake and stomatal opening are necessary to diminish water loss and maintain water potential. Water supplied to climbing and semi-epiphytic plants by aerial roots variously inserted at a number of sites along a stem is evidently sufficient to sustain higher rates of CO
2
exchange and water loss.
P0579 – ePoster
How nitrogen and phosphate deficiency affect strigolactone production and plant growth
Yoneyama, K
Yokota, T 3
1 , Xie, X
, Yoneyama, K
1
1
, Nomura, T 1 , Sekimoto, H 2 ,
1
2
Weed Science Center, Utsunomiya University, Japan;
3
Faculty of Agriculture, Utsunomiya University, Japan;
Dept of Biosciences, Teikyo University, Japan
Strigolactones (SLs) function as a germination stimulant of root parasitic weeds and a hyphal branching factor of
AM fungi in the rhizosphere, and also as a plant hormone inhibiting shoot branching in planta. In this study, effects of nitrogen (N) and phosphate (P) deficiency on SL production and plant growth were examined in Fabaceae
(alfalfa and Chinese milk vetch), Asteraceae (marigold and lettuce), Solanaceae (tomato), and Poaceae (wheat) plants. In alfalfa and tomato, only P deficiency promoted
SL exudation, by contrast, in Chinese milk vetch, lettuce, marigold and wheat, N as well as P deficiency enhanced it. N and P deficiency negatively affected shoot fresh weight and positively affected root length in all plants examined. Root fresh weight responses to N and P deficiency varied with plant species. Loss of these nutrients did not affect root fresh weight in alfalfa and wheat, increased in Chinese milk vetch, and suppressed in lettuce, marigold and tomato. Interaction of nutrient deficiency and SL production will be discussed.
P0584 – ePoster
Effects of drought stress produced by PEG-6000 on antioxidant enzymes activity of two sensitive and
522 tolerant cultivars of Safflower (
Carthamus tinctorious
1
L.)
Zahraeifard, S 1 , Kholdebarin, B 1
Shiraz University, Iran
, Moradshahi, A 1
In order to study the adaptability of Safflower
( Carthamus tinctorius L.) plants to water deficit, we analyzed the activity of antioxidant enzymes, including guaiacol peroxidase ( GPX ,EC 1.11.1.7) and Catalase (
CAT ,EC 1.11.1.6), in Safflower shoots and roots subjected to drought stress during seedling growth stage.
The germination rate and shoot dry weight of five safflower cultivars were studied under different osmotic potentials (0, -0.05, -0.1, -0.5, -1.0 MPa) prepared by polyethylene glycol 6000 (PEG-6000). Safflower cultivars, IL111 and Mahali Esfahan were selected as stress–tolerant and stress–sensitive, respectively, and were used for further characterization. After PEG-6000 treatment, IL111 showed higher enzymatic activity of
GPX and CAT in its shoots and roots than Mahali
Esfahan. Guaiacol peroxidase activity remained relatively constant up to -0.5 MPa and increased significantly at -1.0 MPa. Catalase activity increased in both cultivars, but was significantly higher in IL111 cultivar at all osmotic potential tested. Since biotic and abiotic stresses cause increase in reactive oxygen species
(ROS) which reduce plant growth and development, increased activities of antioxidant enzymes are part of mechanisms which confer tolerance to some varieties of safflower plants such as IL111.
P0585 – ePoster
Micromorphology of secretory trichomes and chemical composition of volatile oils in
Perovskia atriplicifolia
Benth. from north-eastern Romania
Zamfirache, MM
Padurariu, C 1
Mihasan, M 1
1 , Burzo, I
, Olteanu, Z
, Ivanescu, L 1
1
2 , Gostin, I 1
, Badea, ML 2
, Stefan, M 1
, Truta, E 3
1 Alexandru Ioan Cuza University of Iasi, Faculty of
Biology, Romania; 2 Agronomic Sciences and Veterinary
Medicine University of Bucharest, Faculty of
Horticulture, Romania; 3 Biological Research Institute
,
, from Iasi, Romania
Perovskia atriplicifolia Benth. is a plant growing in the rocky regions of Afghanistan and Pakistan, known for its febrifuge properties, for culinary use or its decorative qualities. This species is cultivated in Europe, being also experimentally introduced in Romania as a cultivated plant in order to use its medicinal, aromatic and air purifying (aerosol) properties. In this context, the objective of the present paper is to get thoroughly into the knowledge of the morphological traits and of the distribution rate of the volatile oil–producing secretory trichomes, situated on the surface of the aerial vegetative organs (stems and leaves) and reproductive organs
(flowers) and also to analyze the composition of the volatile oils produced by these organs in anthesis stage, an ontogenetic moment in which the secreted products have real phytoterapeutic and aromatic properties. The micromorphological investigations have been carried out using a surface electron microscopy, at a TESCAN

VEGA II SBH microscope. The volatile oils extraction was conducted using a Clevenger hydrodistillation system. The component separation was performed by gas chromatography, with a 6890 Agilent GC/MS. The volatile compound identification was made using the
NIST spectral bank and Kovats indexes. In the investigated biological material, grown in the pedoclimatic conditions of Moldova region, the secretory trichomes are multicellular structures, with a specific conformation, depending on the analyzed plant part. The number of volatile compounds identified in the oil samples is variable, according to the analyzed organ. The obtained data contribute to the best understanding of the biology of some aromatic and medicinal species of real practical interest, originated in different geographical regions and cultivated in the pedoclimatic conditions of
Romania.
P0587 – ePoster
Epicuticular wax of
Prunus laurocerasus
does not establish the transpiration barrier of the cuticle
Zeisler, VV 1 , Schreiber, L 1
1 Bonn University, Germany
Leaves are covered by a thin extracellular layer, the plant cuticle, limiting the transpirational water loss, reflecting
UV-radiation and protecting leaves from infection by pathogens. The cuticle consists of the insoluble polymer cutin and soluble cuticular lipids also called wax. Wax can be found embedded in the polymer and on the outer surface of the cuticle, where they can form characteristic epicuticular wax crystals or an epicuticular wax film as it is the case with Prunus laurocerasus . Cuticular wax of
Prunus is composed of linear long-chain aliphatics, e.g. alkanes, acids, aldehydes and esters and triterpenoids representing the most abundant substance class. It has been shown that triterpenoids are nearly exclusively found in the intracuticular wax fraction, whereas, linear long-chain aliphatics occur as epi-and intracuticular wax.
It is well known that the transpiration barrier of the cuticle is established by cuticular wax, since upon wax extraction rates of cuticular transpiration increase by 2-3 orders of magnitude. However, it is still an unsolved question, whether the cuticular transpiration barrier formed by wax, is established by the epicuticular, the intracuticular or both wax fractions. Using collodion, epicuticular wax was mechanically removed from the surface of the isolated Prunus cuticle and the intact
Prunus leaf and thus, successfully separated from intracuticular wax without cuticle damage. Selective wax removal was visualized and verified by scanning electron microscopy. The appearance of the cuticular surface was smooth after 2 consecutive treatments with collodion indicating a complete and successful removal of epicuticular wax. Amount and composition of isolated wax was quantified and identified by gas chromatography and mass spectrometry. The chemical analysis clearly indicated that the first 2 collodion strips were nearly exclusively containing linear long-chain aliphatics. Triterpenoids and the remaining fraction of aliphatics localized in the cutin polymer, were only obtained upon a complete wax extraction using organic solvent. Using radio-labelled water, cuticular transpiration of isolated cuticles and intact leaf disks was quantitatively measured before and after selective removal of epicuticular wax. Selective removel of epicuticular wax did not affect rates of cuticular transpiration. This allows to conclude that the epicuticular wax layer of the Prunus cuticle does not establish or significantly contribute to the formation of the cuticular transpiration barrier, which must be formed by intracuticular wax. In current experiments cuticular transpiration before and after collodion treatment of further species is investigated, in order to test whether this observation made for Prunus also applies to other species.
P0588 – ePoster
Leaf and stem structural traits of Patagonian shrubs impact more freezing resistance and plant water relations than habitat characteristics
Zhang, Y-J
Arias, NS 3
1, 2 , Bucci, SJ 3
, Cao, K-F 1
, Hao, G-Y
, Goldstein, G 4
4 , Scholz, FG 3 ,
1 Xishuangbanna Tropical Botanical Garden, CAS,
Mengla, China;
USA; 3
2 University of Miami, Coral Gables,
Universidad Nacional de la Patagonia San Juan
Bosco, Comodoro Rivadavia, Argentina; 4 Universidad de
Buenos Aires, Buenos Aires, Argentina
Plant water relations characteristics and freezing resistance of six Patagonian shrub species from a high elevation plateau site with relatively low temperatures
(about 700 m asl) and a low elevation site, close to sea level, with milder low temperatures were studied in an attempt to understand their freezing resistance mechanisms as well as the relationships among leaf and stem structures, water relations, freezing of plant tissues, and temperature regime of the habitat. Only one species growing in the low elevation site and with the lowest leaf ice nucleation temperature (-8.4°C), avoided extracellular freezing by permanent supercooling. The rest of the species were able to tolerate extracellular freezing from -
8.2 to -20°C. No significant differences were found between low elevation and plateau shrubs in wood density, specific leaf area, leaf and stem hydraulics, leaf ice nucleation temperatures, leaf lethal temperatures, and leaf water relations traits, while strong correlations were found across species between plant water relations characteristics and specific leaf area or stem wood density. Correlations were also observed between stem ice nucleation temperature and vessel diameter and between leaf lethal temperature and leaf elastic modulus.
Rigid cell walls helped to avoid or minimize physical damage to cell membranes during extracellular freezing.
Two shrub species with higher wood density (one from the low elevation site and one from the plateau) exhibited substantially lower percent loss of hydraulic conductivities in winter than the other species. These functional relationships related to freezing resistance and water relations of Patagonian shrubs were not habitatspecific. Therefore, leaf and stem structural differences accounted more to plant water relations characteristics and freezing resistances than the habitat specific differences, particularly the temperature regime. Seeding of ice began in the stem xylem and then propagates to leaves; however, no relationship was found between leaf freezing resistance and xylem resistance to embolism
523

caused by freezing, suggesting that these two biophysical processes represent independent adaptations.
P0589 – ePoster
Cell wall modifications in maize
Pulvini
in response to gravitational stress
Zhang, Q 1
Tingey, S
G 1
3
, Pettolino, F
, Beatty, M 3
2 , Dhugga, K 3
, Burton, R 1
, Rafalski, A 3
, Bacic, A 4
,
, Fincher,
1 ARC Centre of Excellence in Plant Cell Walls,
University of Adelaide, Australia; 2 CSIRO Plant
3 Crop Genetics Research Industry, Canberra, Australia; and Development, Pioneer Hi-Bred International Inc.;
4 ARC Centre of Excellence In Plant Cell Walls,
University of Melbourne, Australia
Changes in cell wall polysaccharides, transcript abundance, metabolite profiles and hormone concentrations were monitored in the upper and lower regions of maize pulvini in response to gravistimulation, where maize plants placed in a horizontal position returned to the vertical orientation. Heteroxylan levels increased in the lower regions of the pulvini, together with lignin, but xyloglucans and galactoglucomannan contents decreased. The degree of substitution of the heteroxylan with arabinofuranosyl residues decreased in the lower pulvini, which exhibited increased mechanical strength as the plants returned to the vertical position. No significant changes in measured non-cellulosic wall polysaccharides could be detected on the upper side of the pulvinus, and the cellulose content remained essentially constant in both the upper and lower pulvinus.
Microarray analyses showed that spatial and temporal changes in transcript profiles were consistent with alterations in wall composition observed in the lower regions of the pulvinus. In addition, they also indicated that metabolic pathways leading to the biosynthesis of phytohormones were differentially activated in the upper and lower regions of the pulvinus in response to gravistimulation. Metabolite profiles and measured hormone concentrations were consistent with the microarray data, insofar as auxin and active gibberellic acid concentrations increased in the elongating cells of the lower pulvinus
P0591 – Poster
Molecular analysis of phosphite induced responses in
Arabidopsis thaliana
Berkowitz, O 1,2
O'Brien, PA 1
, Jost, R 2 , Fenske, R 3 , Hardy, GSEJ 1 ,
1 School of Biological Sciences and Biotechnology,
Murdoch University, Murdoch, Australia; 2 School of
Plant Biology, University of Western Australia, Crawley,
Australia; 3 Metabolomics Australia at the University of
Western Australia, Australia
Phosphite (H
2
PO
3
) is a phosphate analog that is not metabolised by plants but can interfere with the phosphate signalling pathway. It inhibits the plant’s phosphate starvation response, e.g. the up-regulation of high-affinity phosphate transporters, and thus has
524 constrictive effects on plant growth under low phosphate supply. Phosphite has also been used widely to protect plants from oomycete pathogens such as Phytophthora and Phytium . Phytophthora species are prominent pathogens in agriculture, e.g. Phytophthora infestans being the causing agent of potato blight (Irish potato femine). Phytophthora cinnamomi has devastating effects
('dieback disease') on native ecosystems with over 3000 plant species at risk in Western Australia alone.
Phosphite is the only known protectant of plants and exhibits a complex mode of action. At elevated concentrations it directly inhibits the pathogen’s growth by interference with its phosphate-dependent metabolism and/or phosphate signalling which is paralleled in plants grown on high phosphite concentrations. In addition to these direct effects phosphite also induces some of the plant’s defence responses, e.g. treatment of plants leads to increased expression of defence genes. However, the underlying mechanism of this indirect effect is not understood. We have started to characterise the impact of phosphite on plant defence responses by analyses of phenotypic changes, gene expression and metabolic pathways in the model plant species Arabidopsis thaliana . Transgenic plants have been generated that express a microbial phosphite dehydrogenase which converts phosphite into phosphate. These plants are a valuable tool to dissect direct from indirect phosphite effects.
P0592 – Poster
Lateral branching oxidoreductase, a new player in the strigolactone branching pathway that acts downstream of MAX genes but upstream of auxin feedback
Brewer, PB
Frickey, T 2
1 , Meyers, E 1
, Filado, F 1
, Dun, EA 1
, Cremer, J 1
, Mason, M 1
, Weiler, G 2
1
Beveridge, CA 1
University of Queensland, School of Biological
Sciences, St Lucia, Australia; 2 Australian National
University, Research School of Biological Sciences,
,
,
Canberra, Australia
LATERAL BRANCHINGOXIDOREDUCTASE (LBO) was isolated using a microarray and reverse genetics approach to find genes co-regulated with MAX strigolactone (SL) biosynthesis genes. Mutants for lbo have intermediate branching, which is fully rescued by
SL application. Grafting and double mutant analysis suggest action downstream ofMAXstrigolactonebiosynthesis genes and upstream of
MAX2 which is likely involved in strigolactone response. Auxin feedback and enhanced auxin transport are not observed in lbosingle and lbo max double mutants, suggesting that a product of LBO action may also function as a trigger for auxin feedback when SL signalling is reduced. LBO expression is feedback regulated, but not auxin regulated, presumably avoiding an otherwise positive feedback loop with auxin. Further study of LBO will provide great inroads into unravelling
SLproduction, transport, bioactivity and feedback, and provide and new tool for dissecting strigolactone and auxin action in plants.

P0593 – Poster
Salinity-triggered crassulacean acid metabolism induction in
Mesembryanthemum nodiflorum
plants results in differential expression of two carbonic anhydrase isoforms
Clayton, H 1 , Barrett-Lennard, E 2 , Ludwig, M 3
1 School of Biomedical, Biomolecular and Chemical
2
Sciences, University of Western Australia, Australia;
School of Plant Biology, University of Western
3 Australia, Australia; Dept of Agriculture and Food
Western Australia, Centre for Ecohydrology, University of Western Australia, Australia
Mesembryanthemum nodiflorum L. (Aizoaceae; the slender ice plant) is a member of a group of plants collectively called C
3
–crassulacean acid metabolism
(CAM) intermediates. These plants are capable of switching between two pathways of carbon assimilation, namely C
3
photosynthesis and CAM, in response to changing environmental conditions. M. nodiflorum , a halophytic annual, switches from C
3 photosynthesis to
CAM in response to increased soil salinity or reduced water supply, a water-conserving adaptation that makes this plant a highly successful weed in agricultural regions of Western Australia and responsible for significant financial losses to pastoralists and crop farmers. In this study, CAM induction was triggered in glasshousegrown M. nodiflorum plants, as indicated by approximately ten-fold diurnal fluctuations in shoot tissue acidity, by daily watering with saline solution for five weeks. Compared to well-watered control plants, which remained in C
3 photosynthesis mode, shoot tissues of CAM-performing M. nodiflorum plants had eight-fold higher shoot tissue malate concentrations at dawn. The switch to CAM was accompanied by increased expression of the essential CAM enzyme phosphoenolpyruvate carboxylase (PEPC), slightly decreased expression of ribulose-1,5-bisphosphate carboxylase oxygenase (Rubisco), which has a role in both C
3
photosynthesis and CAM, and differential expression of two isoforms of carbonic anhydrase (CA), the enzyme which supplies inorganic carbon to both
PEPC and Rubisco, albeit in different forms. Using a
PCR-based approach, two cDNAs encoding distinct CA isoforms were isolated from M. nodiflorum shoot tissue, and bioinformatic analyses suggested that the two isoforms may be found in different intracellular compartments, consistent with the activities of PEPC and
Rubisco. Quantitative reverse transcription-PCR analysis showed that transcript abundance of the putative cytosolic CA was twice as high in shoot tissue of M. nodiflorum plants using CAM versus C photosynthesis,
3 while the abundance of the putative chloroplastic CA was approximately 50% less. The data support a model in which salinity-triggered CAM induction in M. nodiflorum is accompanied by downregulation of a chloroplastic CA that has a role in C
3
photosynthesis, and significant upregulation of a cytosolic CA that functions to supply HCO
3
to PEPC during CAM.
P0594 – Poster
Antagonistic action of strigolactone and cytokinin in bud outgrowth control
Dun, EA 1 , De Saint Germain, A 2
Beveridge, CA 1
, Rameau, C 2 ,
1 University of Queensland, School of Biological
Sciences, St Lucia, Australia; 2 Institut Jean-Pierre
Bourgin, INRA UMR1318 INRA-AgroParisTech,
Versailles, France
Cytokinin has long been implicated as a promoter of bud outgrowth in plants, but exactly how this is achieved in coordination with other plant hormones was unclear. The recent discovery of strigolactones as the longsoughtbranch inhibiting hormone allowed us to test how cytokinin and strigolactones co-ordinately regulate bud outgrowth in garden pea. We found that strigolactone deficient plants are more sensitive to stimulation of bud growth by cytokinin than wild-type plants, and that buds of wild-type plants are comparativelyresistant to stimulation by cytokinin supplied to the vasculature.
When the hormones were supplied locally or in the xylem stream, exogenous strigolactone was able to reduce the effects of exogenous cytokinin on bud outgrowth. These data suggest that strigolactones do not affect the delivery of cytokinin to axillary buds, and vice versa. Rather, these data combined with dose-response experiments suggest that strigolactones and cytokinin act antagonistically on bud outgrowth, potentially converging at a common point in the bud outgrowth regulatory pathway. Indeed, we found that expression of
PsBRC1, a TCP transcription factor thought to act downstream of strigolactones, is regulated by both cytokinin and strigolactone in a direct manner that does not require protein synthesis, and that expression of
PsBRC1 shortly after treatment correlates with observed bud growth responses.
P0595 – Poster
Effects of the GA insensitive mutation Vvgai1 on the underlying components of relative growth rate in
1 grapevine
Edwards, E 1 , Güsten, S 2 , Thomas, M
CSIRO Plant Industry, Australia; 2
1
University of Applied
Sciences, Geisenheim, Germany
Gibberellins (GAs) have long been known to promote growth and mutants that are either deficient in GA or insensitive to GA are typified by a dwarf stature. The previously described GA insensitive grapevine mutant, known as the 'microvine', has such a stature due to possessing much shorter internodes than the wildtype, but also produces inflorescences in place of tendrils.
Such changes in morphology are likely to have an impact on carbon partitioning within the vine as well as relative growth rate and its underlying components. Furthermore, there are conflicting reports of the effects of exogenously applied GA on leaf level photosynthesis and GA insensitive mutants provide another mechanism whereby the potential role of GAs in altering photosynthesis can be studied. Using green cuttings we have examined
525

carbon allocation, growth rate and photosynthetic rate of the microvine and wildtype. Specific leaf area (SLA), leaf mass ratio (LMR), leaf level assimilation under saturating light (Asat), and assimilation per unit leaf nitrogen, parameters positively associated with growth rate, were all higher in the microvine than wildtype, yet relative growth rate of the microvine was lower than the wildtype. Consequently, net assimilation rate (NAR) must have been significantly reduced in the microvines and was estimated as only 50% of the wildtype NAR.
Potential explanations for this scenario (high LMR, SLA and Asat but low NAR) will be discussed, but include
Asat not being representative of actual whole plant assimilation, due to a high degree of self shading for example, and high rates of respiration. Other aspects of carbon allocation were also affected, with the microvines having a reduced fraction of biomass in the stem, compared with wildtype vines of the same total mass, and an increased fraction in the root system.
P0596 – Poster
The first chlorogenic acid ester saponin from
Lonicera macranthoides
Hand.-Mazz.
Feng, X 1
Dong, Y 1
, Chen, Y 1 , Wang, M 1 , Zhao, Y 1 , Sun, H 1 ,
1 Institute of Botany, Jiangsu Province and Chinese
Academy of Sciences, Nanjing, PR China
The dried flower buds of Lonicera macranthoides Hand.-
Mazz. (Caprifoliaceae) are commonly used traditional
Chinese medicines in the southwest of China. It has been utilized in China as an antipyretic and detoxicant agent, and has been widely used to treat carbuncle, throat obstruction, erysipelas, fever and cold in clinical and civil. Although this species had been broadly used, the study on its chemical constituents is still empty. To understand this new species better, systematical studies on chemical constituents of L. macranthoides were carried out. 41 compounds were isolated from the ethanol extract of flower buds of L. macranthoides , including 21 triterpenoid saponins, 5 caffeoylquinic acid derivatives, 8 flavonoids and so on. Throw in vitro and in vivo using cell proliferation assays, two saponins (Macranthoside B and Lonimacranthoide I) show remarkably anti-tumor activity. Herein we report the isolation and structural elucidation of the new saponin named Lonimacranthoide
I (1), which is a rare chlorogenic acid ester acylated on the C-23 of hederagenin. The flower buds of L. macranthoides , were collected from Hunan province of
PR China. The dried buds (10.0kg) were extracted with hot 95% ethanol for three times. After removal of ethanol, the residued water suspension was re-extracted with petroleum ether, EtOAc respectively and the obtained aqueous portion was passed through Diaion HP-
20 and eluted with water, 50% EtOH and 90% EtOH.
The 50% EtOH fraction (30.0g) was chromatographed on silica gel columns using a gradient of CHCl3: MeOH:
H2O (17:3:0.2 –> 4:1:0.1 –>7:3:0.5 –>3:3:0.5) to give 9 fractions (A-I). The fraction I (2.0g) was repeatedly chromatographed on RP-C18 and Sephadex LH-20 columns using an MeOH-H
2
O solvent system to give compounds 1 (20mg). 1 was obtained as an amorphous powder. The molecular formula was established to be
C
81
H
122
O
40
by its positive HR-ESI-MS data and DEPT
526
NMR spectra. The spectral features and physicochemical properties revealed 1 to be a triterpenoid saponin. The information of NMR spectrum indicated that skeleton of
1 is hederagenin. The monosaccharides were identified as arabinose, rhamnose and glucose (ratio 1:1:4) by GC analysis with authentic monosaccharides and a combination of 2D NMR experiments. The sequence of the sugar linkages connected to the aglycone was deduced from the following HMBC correlations. Besides the signals due to the aglycone and component sugars, the 1H\13C NMR spectra of 1 showed the presence of the chlorogenic acid moiety. Comparison of the 13C
NMR spectra of 1 with those of macranthoidin B showed that the chlorogenate group was assigned to attach at C-
23 (
δ
66.5) of the aglycone. The HMBC spectrum provided further confirmation of this chlorogenic acyl group from the correlation between H-23 (
δ
4.40, 4.80) of the aglycone and C-7'(
δ
174.2) of the chlorogenic acyl group. On the basis of the above evidences,
Lonimacranthoide I (1) was found to be a new compound and established as 3-O-
β
-D-glucopyranosyl-(1–>4)-
β
-Dglucopyranosyl-(1–>3)-
β
-L-rhamnopyranosyl-( 1–>2)-
β
-
L-arabinopyranosyl hederagenin 23-O-chlorogenic acyl-
28-O-
β
-D-glucopyranosyl- (1–>6)-
β
-D-glucopyranosyl ester.
P0597 – Poster
Flavonoid diversity of
Saussurea species
(Asteraceae) in the Altai Mountains
1
Iwashina, T 1 , Smirnov, SV 2
K 4
, Damdinsuren, O 3 , Kondo,
Dept of Botany, National Museum of Nature and
2 Science, Japan; Dept of Plant Systematics, Altai State
University, Russia; 3 Biological Division, Hovd Stae
University, Russia; 4 Deopartment of Agriculture, Tokyo
University of Agriculture, Japan
The genus Saussurea DC. (Asteraceae) consists of ca.
300 species, which are mainly distributed in East Asia.
Some of the plants occur in the Himalayas, the Tien Shan
Mountains and the Altai Mountains. It has been reported that the Saussurea is morphologically and geographically diversified. However, their chemical compounds are hardly analysed. We have surveyed nine Saussurea species, S. alberti, S. elegans, S. gnaphalodes, S. involucrata, S. konuroba, S. kuschakewiczii S. leucophylla, S. schanginiana and S. sordida in Tien Shan
Mountainsin Kazakhstan and Kirgyz for flavonoids. Ten flavonol glycosides, kaempferol 3-O-rutinoside, 5-Oglucoside and 7-O-glucoside, quercetin 3-O-rutinoside,
3-O-glucoside, 3-O-galactoside, 5-O-glucoside and 7-Oglucoside, and isorhamnetin 3-O-rutinoside and 5-Oglucoside; eight flavone aglycones, luteolin, luteolin 7methyl ether, hispidulin, nepetin, selagin, selagin 7methyl ether, velutin and jaceosidin; and nine flavone Oglycosides, apigenin 7-O-rutinoside and 7-O-glucoside, luteolin 7-O-rutinoside, 7-O-glucoside, 7-O-galactoside,
7-O-glucuronide and 5-O-glucoside, hispidulin 7-Oglucoside and nepetin 7-O-glucoside, and four Cglycosylflavones, isovitexin, isoorientin, apigenin 6,8-di-
C-glycoside and luteolin 8-C-glycoside, were found in their species in various combination (Kusano et al.,
2007). As a series of flavonoid survey in the genus
Saussurea , flavonoids of the species from the Altai

Mountains and adjacent area were surveyed. Seven
Saussurea species, i.e., S. alpina, S. daurica, S. laciniata,
S. pricei, S. pseudo-alpina, S. salicifolia and S. salsa , were collected in the Altai Mountains in Russia and
Mongolia. Dry leaves or aerial parts were extracted with methanol. After concentration, flavonoids were isolated by preparative paper chromatography and then purified by Sephadex LH-20 column chromatography.
Identification was performed with UV, LC-MS, acid hydrolysis, and direct HPLC and TLC comparisons with authentic samples. Thirteen flavonoids, apigenin 7-Oglucuronide and 7-O-glucoside, luteolin 7-O-glucuronide and 7-O-glucoside, chrysoeriol 7-O-glucuronide, 7-Oglucoside and 7-O-rhamnoside, jaceosidin, nepetin, hispidulin as flavones, and quercetin 3-O-rutinoside and
3-O-glucoside and isorhamnetin 3-O-rutinoside as flavonols, were found in various combination. Of their species, S. daurica, S. laciniata and S. salsa synthesized flavone glycosides alone. On the other hand, S. alpina, S. pricei and S. pseudo-alpina occurred flavonol glycosides.
Of three species, S. pricei accompanied with free flavones. S. salicifolia synthesized only flavone aglycone, hispidulin. Thus, it was shown that the genus
Saussurea is not only morphologically and geographically but also chemically diversified.
P0599 – Poster
Functional characterization of
Brassica rapa
GIGANTEA and RNAi-mediated methods in regulation of flowering time
Kim, JA 1 , Kim, WY 2 , Hong, JK 1 , Kim, JS 1 , Lee, Y-H 1
1 National Academy of Agricultural Science, Republic of
Korea;
Korea
2 Gyeongsang National University, Republic of
Flowering is one of the most important development traits for the production of Chinese cabbage ( Brassica rapa ). After planting, sudden low temperature triggers premature flowering leading to a reduction in the yield and quality of harvested production. Therefore, understanding the mechanism of flowering control is important in agronomic practice in preventing Chinese cabbage from flowering prematurely. Mutations in the
Arabidopsis GIGANTEA(GI) gene delay flowering and enlarge the plant size. We isolated the GI gene in
Brassica rapa using in silico method and constructed
RNAi vectors which contained each 10 fragment from the coding region and 3`-UTR. These constructs were transformed into Arabidopsis thaliana L.(Columbia ecotypes). These suppression lines showed various phenotypes in terms of flowering time and size. The analysis of mRNA expression and phenotypes revealed the domains which showed strong suppression effects on
GI expressions. In this study, we confirmed function of
BrGI in flowering time and plant size and also suggested the new possibility to regulate flowering time of B. rapa. by the RNAi-mediated knockdown.
P0600 – Poster
Pigment components in the flowers of Dutch
Iris cultivars and their contribution to the flower color
Mizuno, T 1 , Yabuya, T 2 , Kitajima, J 3 , Iwashina, T 4
1 Graduate School of Agriculture, Ibaraki University,
Japan;
Japan; 3
2 Faculty of Agriculture, Miyazaki University,
Showa Pharmaceutical University, Japan; 4 Dept of Botany, National Museum of Nature and Science,
Japan
Dutch iris ( Iris × hollandica ), which belongs to the bulbous Iris group, is ornamentals in the family
Iridaceae. There are blueish flower cultivars in this Iris , besides yellow, white and violet flowers. Although bluish flowers of this iris may be has highest value among ornamentals of all Iris species and had conducted the molecular genetic characterization of pigment biosynthesis, information of pigment components is unsufficient. In this study, we have carried out to obtain the data of pigment components using blueish and purple flowers of two cultivars, ‘Blue Diamond’ and ‘Teru-
Murasaki’. In addition, we discussed the cause of bluing effect in perianths by the qualitative and quantitative comparisons of pigment components between two cultivars. As the flower pigments, 12 anthocyanins were isolated and characterized. Of their anthocyanins, six ones are the acylated delphinidin glycosides such as delphanin (delphinidin 3-O-(p-coumaroylrutinoside)-5-
O-glucoside), and were detected in both cultivars. Other six anthocyanins are acylated petunidin and malvidin glycosides such as petanin (petunidin 3-O-(pcoumaroylrutinoside)-5-O-glucoside) and ensatin
(malvidin 3-O-(p-coumaroylrutinoside)-5-O-glucoside).
They were detected in purple flower cultivar ‘Teru-
Murasaki’ but not in ‘Blue Diamond’. Thus, it was presumed that the difference of flower color is due to the qualitative differences of anthocyanins between two cultivars. Other flavonoids were isolated and characterized as C-glycosylflavones i.e. isovitexin, isoorientin and swertiajaponin etc, and their Oglycosides, which were presumed to be co-pigment substances. Furthermore, of isolated sixteen flavones, four ones were acylated with acetic acid, and may be new flavones. By quantitative HPLC comparisons of the floral extracts of two cultivars, total anthocyanin content
(TA) of ‘Teru-Murasaki’ was three times than that of
‘Blue Diamond’. On the other hand, total flavone content
(TF) of ‘Blue Diamond’ was three times than that of
‘Teru-Murasaki’. Co-pigment index (TF/TA) was calculated and that of ‘Teru-Murasaki’ was revealed eight times higher than that of ‘Blue Diamond’. Thus, it was indicated that the flavones contribute as bluing substances. There is also significant difference in contents of the acetylated flavones between ‘Blue
Diamond’ and ‘Teru-Murasaki’. Though total acylated flavone content in ‘Blue Diamond’ were 34%, that in
‘Teru-Murasaki’ was only 1.8% of total flavones. Thus, it was indicated that acetylated flavone C-glycosides strongly contribute as co-pigment substances. In conclusion, we presume that the bluish flowers of Dutch
Iris cultivars effectively occur as the result of intramolecular co-pigmentation of each acylated flavonoids, i.e. malonylated anthocyanins and acetylated flavones.
P0601 – Poster
Interaction between carbon and nitrogen metabolisms in the C
3
–CAM epiphytic bromeliad
Guzmania monostachia
527

1
Pereira, PN 1 , Freschi, L 1 , Mercier, H 1
Dept of Botany, Institute of Biosciences, University of
São Paulo, São Paulo, SP, Brazil
Guzmania monostachia is an epiphytic bromeliad that can shift from C photosynthesis to Crassulacean acid
3 metabolism (CAM) in response to changes in the environmental conditions. When subjected to water stress, this bromeliad has been shown to perform CAM photosynthesis exclusively at the apical leaf portion, whereas, in contrast, its basal leaf region displays typical
C3 photosynthesis. Previous data obtained in our laboratory have demonstrated that when incubated for 7 days in the presence of 30% polyethylene glycol (PEG), adult plants of G. monostachia switch from C
3
to CAM, exhibiting the highest levels of titratable acidity and activity of the enzymes phosphoenolpyruvate carboxylase (PEPC)and malate dehydrogenase (MDH) specifically at the upper leaf portion. Taking advantage of this plant model, the present study attempted to investigate a possible relationship between the carbon and nitrogen metabolisms throughout the leaf blade of this bromeliad. To achieve this, detached leaves of 2year-old individuals of G. monostachia were subjected to a treatment with 30% PEG for 7 days or were maintained in water for a similar period (control). Subsequently, the nighttime accumulation of titratable acidity and the activities of PEPC and MDH were determined in the apical, middle and basal leaf portions in order to estimate the occurrence of CAM photosynthesis in these leaf regions. Moreover, the diurnal rhythm of in vivo activities of nitrate reductase (NR) and glutamine synthetase (GS) as well as the day/night pattern of organic acids (malate and citrate) levels were also determined. The results showed that CAM photosynthesis can be induced in G. monostachia leaves within 7 days of water deficit (PEG) treatment, as evidenced by the increased nocturnal accumulation of titratable acidity and activities of PEPC and MDH detected in the upper leaf portion of this bromeliad.
Interestingly, the day/night pattern of NR activity revealed that nitrate reduction in G. monostachia occurs mainly during the night either in individuals performing
CAM (water deficit treatment) or in plants displaying C
3 photosynthesis (control condition). The diurnal pattern of
GS activity and the nighttime accumulation of malate and citrate were also determined in these same leaf regions and treatments, providing further evidence of a possible relationship between carbon and nitrogen metabolisms in this epiphytic bromeliad.
P0602 – Poster
Diurnal changes in endogenous cytokinins, organic acids and starch in the C
3
-CAM epiphytic bromeliad
Guzmania monostachia
Pereira, PN 1 , Freschi, L 1 , Mercier, H 1
1 Dept of Botany, Institute of Biosciences, University of
São Paulo, São Paulo, Brazil
Previous results obtained in our laboratory have demonstrated that different leaf sections of the tank C3-
CAM bromeliad Guzmania monostachia can perform distinct photosynthetic behaviors. When challenged by a
528 restriction in water availability, the upper leaf section of this bromeliad switch from C3 photosynthesis to
Crassulacean acid metabolism (CAM), exhibiting considerably higher levels of nocturnal titratable acidity and extractable activities of the CAM-related enzymes phosphoenolpyruvate carboxylase (PEPC)and malate dehydrogenase (MDH) than those detected at basal and middle leaf regions. Based on the limited literature data currently available on the endogenous signaling networks controlling CAM expression and taking advantage of the high photosynthetic plasticity exhibited by G. monostachia plants, the present work attempted to investigate a possible influence of endogenous cytokinins in the regulation of this photosynthetic pathway. To achieve this, detached leaves of G. monostachia were subjected to a treatment with 30% PEG for 7 days or were maintained in water for a similar time period
(control). Subsequently, the occurrence of CAM photosynthesis in the apical, middle and basal leaf portions was evaluated by measuring the nighttime accumulation of titratable acidity and the activities of
PEPC and MDH. In addition, the endogenous levels of four cytokinin species, zeatin (Z), Z riboside (ZR), isopenteniladenine (iP) and iP riboside (iPR) were determined every 3h throughout the day/night cycle.
Samples collected at these same harvesting points were also analyzed for their levels of organic acids (malate and citrate), soluble sugars (sucrose, glucose, fructose) and starch in order to characterize the daily carbon fluxes in this bromeliad. Our results revealed that the water-stresstriggered CAM induction in G. monostachia can occur within 7 days of treatment, since the highest levels of nocturnal acid accumulation and PEPC and MDH activities were detected at the 7th day of water stress treatment. The results of day/night fluxes of organic acids, soluble sugars and starch confirmed the occurrence of a typical CAM cycle in water-stressed plants.
Comparisons between the diurnal changes in endogenous cytokinins and the detailed data obtained about the CAM cycle, allowed us to hypothesize a possible relationship among hormonal signaling (cytokinins), CAM metabolism and the production of specific carbohydrates and organic acids throughout the day/night cycle. A possible interplay between the endogenous levels of cytokinins and the water-stress-induced CAM expression in G. monostachia will also be discussed
P0603 – ePoster
Evaluation of P5CS gene over expression on protein pattern variations in transgenic tobacco plants under in vitro salt stress
Razavizadeh, R 1 , Ehsanpour, AA 2
2
1 Biology Dept, Payame Noor University, Tehran, Iran;
University of Isfahan, Isfahan, Iran
Drought and salinity are important osmotic and ionic stresses that limit plant growth and crop productivity . In this study, protein pattern changes of transgenic tobacco plants carrying P5CS gene (a key enzyme in the proline biosynthetic pathway) under salt stress were evaluated.
Transgenic and non transgenic tobacco plants were grown on MS medum containing 0, 150, 300 mM NaCl for 2 days under in vitro culture condition and then protein patterns of leaves and roots were analyzed by
SDS-PAGE and Image J program. Up regulated and

down regulated protein bands in leaves and roots showed remarkable changes of proteins expression between transgenic and non transgenic as well as salt treated and untreated plants. Proteome analysis of proteins by twodimensional polyacrylamide gel electrophoresis (2DE) showed expression level of 8 proteins including proteins involved in photosynthesis, defense, metabolism were significantly changed under salt stress.
P0604 – Poster
The anthocyanins in purple and blue
Clematis cultivars and their contribution to the flower colors
Sakaguchi, K 1 , Kitajima, J 2 , Iwashina, T 3
1 United Graduate School of Agricultural Science, Tokyo
University of Agriculture and Technology, Japan;
2 Showa Pharmaceutical University, Japan; 3 Dept of
Botany, National Museum of Nature and Science, Japan
The genus Clematis belongs to the family Ranunculaceae and ca. 3,000 cultivars have been bred until now.
Recently, the flower pigments of some red and yellow
Clematis species and cultivars were reported. In red flowers, six polyacylated anthocyanins based on cyanidin have been isolated and identified. On the other hand, in yellow flowers, it was shown that much amounts of flavonols, especially quercetin glycosides such as 3-Oglucoside, 3-O-galactoside and 3-O-rutinoside, were accumulated as flower pigments in the sepals. However, the pigments of purple and blue Clematis flowers and mechanism of pigmentation have hardly been surveyed.
In this study, we describe the identification of the major anthocyanins of purple and blue Clematis cultivars and discuss the flower color expression of purple and blue flowers. Two Clematis cultivars, ‘Jackmanii Superba’ and ‘Fujimusume’, were used as plant materials for isolation and identification of the anthocyanins. The flower color of the former cultivar is purple and that of the latter one is blue. These plant materials are cultivated in Tsukuba Botanical Garden, National Museum of
Nature and Science, Tsukuba, Japan. The fresh sepals were extracted with HCOOH/MeOH(8:92) and were applied to Amberlite XAD-7 column chromatography.
After concentration, crude anthocyanins were applied to prep. PC and purified by Sephadex LH-20 column and finally prep. HPLC. The isolated anthocyanins were identified by UV spectra, saponification, partial and complete acid hydrolysis, LC-MS, and 1H and 13C
NMR including H-H COSY, C-H COSY, NOESY,
HMBC and HSQC. These anthocyanins consisted of cyanidin, petunidin and delphinidin and ca. 80% were delphinidin glycosides. They were attached glucose and/or galactose and glucuronic acid as glycosidic sugars, and caffeic acid, malonic acid or succinic acid as organic acids in various combination. Of their anthocyanins, one of the major anthocyanins in
‘Jackmanii Superba’ was identified as delphinidin 3-O-
[2-O-(2-(trans-caffeoyl)-
β
-glucopyranosyl)-(6-succinyl)-
β
-galactopyranosid e]-7-O-
β
-glucopyranoside. The remained ones were characterized as delphinidin 3-Oglucosylgalactoside-7-O-glucoside which was acylated with caffeic acid, malonic acid and aromatic acid of molecular weight 312 from ‘Jackmanii Superba’ and delphinidin 3-O-glucosylgalactoside-3'-O-glucuronide acylated with caffeic acid, malonic acid and aromatic acid of molecular weight 312 from ‘Fujimusume’. The absorption maxima of the antocyanin in the former cultivar showed 543 nm. On the other hand, that of the latter cultivar was 535 nm. Therefore, we presumed that the intramolecular copigmentation occurred between aromatic acid and anthocyanidin nuclei. Furthermore, the flower color of ‘Fujimusume’ was more bluish than that of ‘Jackmanii Superba’. We presumed that flower color of ‘Fujimusume’ was expressed by not only the color of anthocyanins themselves but also other factors, e.g., the intermolecular copigmentation between anthocyanins and other flavonoid such as flavonols. The total anthocyanin content in ‘Fujimusume’ flowers is ca. 14 times less than that of ‘Jackmanii Superba’. Therefore, we presumed that
‘Fujimusume’ is more bluish because the efficiency of the intermolecular copigmentation is higher than
‘Jackmanii Superba’. We will perform in vitro reconstruction of the anthocyanin complex, which consists of anthocyanin and copigment.
P0605 – Poster
Morphological characteristics and pigmentary variation in the progenies from Nagai type of
Japanese garden Iris (
Iris ensata
Thunb.)
Sakai, K 1 , Murai, Y 2 , Bang, SW 1 , Iwashina, T 3 , Kaneko,
Y 1
1 Laboratory of Plant Breeding, Faculty of Agriculture,
Utsunomiya University, Japan; 2 Dept of Chemistry, Keio
University, Japan; 3 Dept of Botany, National Museum of
Nature and Science, Japan
Japanese garden iris, Iris ensata , is developed from wild form, Iris ensata var. spontanea , and it is classified into four groups such as Nagai, Edo, Ise, and Higo types, consisting of more than 2000 cultivars. Especially, Nagai type is bred from wild form in Mt. Iide. It is valuably conserved 58 cultivars in the Nagai city, Yamagata Pref.,
Japan However, there aren’t so much historical references and genetical information.So, to investigate
Nagai type’s potential for iris breeding, I studied on the morphological characteristics in F1 and F2 progenies from Nagai type such as ‘Tsumabeni’, ‘Usubeni’ and
‘Mifuchi-no-nagare’, and analyzed the anthocyanins in their inner and outer perianths of flowers by HPLC.
(1)F1 plants of ‘Tsumabeni’ and ‘Usubeni’ were morphologically classified into eight groups. On the other hand, F1 plants of ‘Mifuchi-no-nagare’ were morphologically classified into three groups. (2) The F2 from F1 plants of ‘Tsumabeni’ showed some variations in the form and color. In addition, flower color such as
‘sunago’ and ‘kasuri’ unexpectedly expressed in F2.
Moreover, the dwarf plants were also detected from one of the eight groups. (3) Flowering time in three cultivars of Nagai type and their F1 plants were from the end of
May to the beginning of June. Their pollen fertility was high. (4)The anthocyanins in the F1 plants were classified into three types; type 1 included petanin and ensatin, type 2 is hardly included petanin and ensatin, type 3 is examined faster peak than the other two types or no anthocyanin. The anthocyanin of ‘Tsumabeni’ and
‘Usubeni’ were type 1, ‘Mifuchi-no-nagare’ was type 3.
Consequently, Nagai type is useful genetic resources for the breeding of I. ensata.
529

P0606 – Poster
Characterization of tomato DREB2 family genes encoding stress-responsive transcription factors
Sakuma, Y 1
K 1
, Miura, Y
, Matsuda, S 1
1 , Hara, H
, Inoue, M 1
1 , Mamesaya, R 1 , Seike,
1 Ehime University, Japan
Environmental stresses such as heat, cold, drought, and high salinity influence plant growth and productivity.
Plants respond and adapt to these stresses to survive under stress conditions at physiological and biochemical levels. These stresses have been shown to induce the expression of genes with various functions. The DRE containing the core sequence A/GCCGAC was identified as a cis-acting element that is important for the regulation of gene expression in response to drought, high salinity and low temperature stresses in Arabidopsis . cDNAs encoding DRE/CRT-binding proteins, DREB1/CBFs and
DREB2s, have been isolated as trans-acting factors.
These proteins contain APETALA2 (AP2)/ethyleneresponsive element-binding factor (ERF) motifs and specifically bind to DRE/CRT sequences, thereby activating the transcription of genes driven by DRE/CRT sequences. DREB type transcription factors are found not only in dicotyledon but also in monocotyledon such as rice, maize, wheat, barley and rye. Thus, stress responsive mechanism that DREB family proteins are involved with are widely conserved between higher plants. The Arabidopsis DREB1 subgroup consists of six genes and the DREB2 subgroup consists of eight genes.
DREB1A/CBF3, DREB1B/CBF1 and DREB1C/CBF2 are strongly and transiently induced by low temperature stresses. Meanwhile, DREB2A and DREB2B are induced by osmotic and high temperature stresses, thus genes transcription factor may work as pivot of crosstalk of osmotic stress signaling and temperature stress signaling. Regulation of DREB2 transcription factors is complicated. In addition to regulation at transcriptional level, protein stability, alternative splicing and proteinprotein interaction involved with the regulation of
DREB2. And intracellular localization may also be involved with the regulation. Tomato (Lycopersicon esculentum) is important not only as a major vegetable crop, but also as a model plant of Solanaceae family.
Tomato whole genome sequence have already been available that enable us to carry out comprehensive analyses. Therefore, we carried out a whole-genome survey of the tomato DREB2-type genes and found seven members. These seven genes were characterized to determine which of them contribute to plant stress responses.
P0607 – Poster
Flavonoid diversity of some
Chrysanthemum
and
Ajania
Species (Asteraceae) in Japan and China
Uehara, A 1 , Nakata, M 2 , Liang-Sheng, W 3 , Iwashina, T 4
1
2
United Graduate School of Agricultural Science, Tokyo
University of Agriculture and Technology, Japan;
Botanic Gardens of Toyama, Japan; 3 Beijing Botanical
Garden, Institute of Botany, Chinese Academy of
Sciences, China; 4 Dept of Botany, National Museum of
Nature and Science, Japan
Almost all species of the genera Chrysanthemum and
Ajania (Asteraceae) are distributed in East Asia. The former genus consists of 37 species, of which 14 and 17 ones are native to Japan and China, respectively. On the other hand, the genus Ajania includes 34 species in the world, and 4 and 32 species in Japan and China. Both genera can distinguish by morphological characters of the flower heads, i.e. Chrysanthemum put tubular and ligulate flowers and Ajania only have tubular flowers.
However, some authors propose that Ajania is incorporated into Chrysanthemum as a section, because they have similar cytological characters each other and can easily make the hybrids between their species.
Moreover, their molecular phylogeny shows that
Chrysanthemum and Ajania were divided in comparatively recent times, so that it is difficult to decide the phylogeny of their species. In this survey, we tried to clarify and compare the foliar flavonoid characters of
Chrysanthemum and Ajania species native to Japan and
China. Twenty-four species of two genera, i.e. 11 species and 4 varieties, and 3 species from Japanese and Chinese
Chrysanthemum , respectively, and 4 and 2 species from
Japanese and Chinese Ajania , respectively, were used as plant materials. The leaves were extracted with MeOH and their internal flavonoid composition was compared by HPLC. On the other hand, external flavonoids were rinsed with acetone. Their flavonoids were isolated by various chromatography and identified by various spectral survey and so on. External flavonoids of
Japanese species were present as methoxylated flavone aglycones such as jaceosidin and eupatilin, etc., in all species. However, their distribution patterns were apparently irregular among their species. In contrast, those of Chinese ones have the same composition. On the other hand, the general character of internal flavonoids in
Japanese Chrysanthemum species was the presence of flavone and flavanone glucuronides such as luteolin and eriodictyol 7-O-glucuronides. However, in Chinese species of their genera, C. lavandulifolium and C. chanetii , were apparently absent the flavonoid glucuronides except for C. indicum which is also distributed in Japan. Internal flavonoids pattern of
Japanese Ajania species was also different with those of two Chinese species, A. parviflora and A. fruticulosa . In addition, flavonol, quercetin 3-O-rutinoside, which was found in two Japanese species, was absent in Chinese ones. Though, we have surveyed a few Chinese
Chrysanthemum and Ajania species, their flavonoid composition was clearly different with those of Japanese species. Thus, it was indicated that both genera are diversified for flavonoid character. Hereafter, more detailed phytochemical survey will be performed to
Chinese Chrysanthemum and Ajania species.
P1098 – ePoster
Investigating plant-pathogen interactions in wheat
1 root diseases using the model grass
Brachypodium distachyon
Schneebeli, K 1 , Mathesius, U 2 , Harvey, P 3 , Watt, M
CSIRO Plant Industry, Canberra, Australia; 2
1
Research
School of Biology, Australian National University,
530

Canberra, Australia; 3 CSIRO Ecosystem Sciences, Waite
Campus, Glen Osmond, Australia
Brachypodium ( Brachypodium distachyon ) is an emerging model for temperate cereals. In 2010 the genome sequence of the reference line was published.
Sequencing of further lines is underway, to provide a comprehensive genetic resource along with T-DNA insertion lines and mapped recombinant inbred lines
(RILs). The sequenced lines are part of a collection of over 180 natural accessions originating from regions of
Turkey and Spain. These have wide phenotypic and genetic diversity. This project aims to use Brachypodium resources to discover root disease resistance or tolerance mechanisms in wheat. The close phylogenetic relationship between Brachypodium and wheat suggests that genes found for disease tolerance in Brachypodium could reveal orthologous genes in wheat. Brachypodium development is similar to wheat and both species belong to the Pooideae subfamily of the Poaceae. However diploid Brachypodium has a genome size comparable to
Arabidopsis at 272 Mb, while the genome of hexaploid bread wheat is large, complex and unsequenced; around
17,000 Mb. In the laboratory, short-statured
Brachypodium is better suited to root experiments than wheat. Two common Australian wheat root diseases are being used in initial screening, the basidiomycete fungus
Rhizoctonia solani AG8 and the oomycete Pythium irregulare. Both organisms are necrotrophic pathogens, with broad host ranges including cereal, legume and brassica crops. Natural resistance to these pathogens has not been found in wheat. Disease is managed using field management practices and fungicide application. The estimated annual cost of R. solani and Pythium spp. to
Australian wheat production, using current control practices, is $59 million and $10 million respectively.
Assays are being developed to screen for root disease in
Brachypodium. An agar plate assay of 134 natural accessions demonstrated the general susceptibility of
Brachypodium to P. irregulare at germination. Seeds placed on a lawn of Pythium growing on water agar had a lower germination rate and reduced root length compared with the control. Wheat germination was also greatly reduced on the Pythium lawn in this assay. In a pot assay to screen root development in response to R. solani , disease symptoms could be seen in all tested
Brachypodium accessions and wheat. Primary root length is decreased, branch root formation is less symmetric and roots are more prone to breaking. Using total root length and first leaf length as indicators, initial results suggest that Rhizoctonia disease severity may vary between accessions of Brachypodium. A pot assay is also being developed to investigate root development in response to
P. irregulare . The pot assays will be used to screen 180 natural Brachypodium accessions, as well as T-DNA lines and recombinant inbred lines. Lines displaying different responses to the pathogens will be examined more closely, using microscopy and gene expression assays, to understand the tissue and genetic mechanisms of defence or tolerance.
P0609 – Poster
Ultrastructural study of the microorganisms colonizing gametophytes and sporophytes of Canary pleurocarpous mosses
, Ron, E 2
1
Alfayate, C 1
Univ. La Laguna, Fac. Biología, Dpto Microbiología y
Biología Celular, Spain; 2 Dept of Plant Biology, Faculty of Biology, Complutense University , Madrid, Spain
The relationships between microorganisms and bryophytes are not still well known and the bryophytes appear as host plants for the microorganisms in numerous ways (Opelt and Berg 2004). Seven species of pleurocarpous mosses from the Canary Islands laurel forest were found to be colonized by microorganisms and were investigated by LM, SEM and TEM. We report the presence of prokaryotic and eukaryotic microorganisms in extracellular spaces and inside cells of the gametophytes and the sporophytes of Cryptoleptodon longisetus (Mont.) Enroth., Eurhynchium praelongum
(Hedw.) Schimp., Homalothecium sericeum (Hedw.)
Schimp., Isothecium myosuroides Brid., Leucodon canariensis (Brid.) Schwägr., Neckera cephalonica Jur. and Unger., and N. intermedia Brid. Their ultrastructural features resemble be heterotrophic bacteria, cyanobacteria and fungi Ascomycetes. In different regions we detected bacteria crossing the cell wall through plasmodesmata, just as hyphae attack the thickwalled parenchyma of the caulidium, seta and capsules and they could contribute to tissue decomposition.
Interestingly, structures constituted by a phase of resistance surrounded by fungal cells also were observed.
P0610 – Poster
Study of anatomical structure of graft-union in
Leucospermum
(Proteaceae)
Alfayate, C 1 , Gómez, L 2 , Vera-Batista, C 2 , Rodríguez,
1
JA 2
Dept of Microbiology and Cellular Biology, Faculty of
Biology, La Laguna University, Tenerife, Canary
Islands, Spain; 2 Dept of Engineering, Production and
Agricultural Economics, School of Agricultural
Engineering, La Laguna University, Canary Islands,
Spain
This study was aimed primarily to determine the anatomical structure of the graft-union in two graft combinations of Leucospermum (Proteaceae). It was conducted during five periods following grafting, with two different combinations: L.
´Succession I´ / L. patersonii and L . ´Succession I´ / L.
´Spider´, using unrooted cuttings as stocks. Samples were taken from graft-unions, fixed in FAA, embedded in Paraplast Plus and then stained with Safranin-Fast Green (Johansen
1940). It was noted that cells neighbouring the wound
531

were capable of de-differentiation where the two sides come into contact, developing meristematic characteristics and subsequent proliferation. The vascular cambium showed an essential role in engraftment. The cambial bridge was established at 4 months after grafting, and new vascular cambium, phloem and xylem were formed. Cellular activity of both components was necessary to establish the cambial bridge. Xylem tissue was developed regularly in both types of graft, with both
L. patersonii and L.
´Spider´ as stocks, indicating that the conductive elements transported water, essential to keep the grafts alive. Some of the sections of both combinations showed cells with a 'necrosed' appearance in cortical tissues at 5 and 6 months, without negatively affecting compatibility. The combination L.
´Succession
I´ / L . patersonii and L.
´Succession I´ / L.
´Spider´ belongs to the compatibility group 'a' (Mosse and Herrero
1951).
P0611 – Poster
Variations in leaf and stem anatomy of two micropropagated
Leucospermum
(Proteaceae) cultivars
Suárez, E
JF 3
3 , Alfayate, C 1 , Rodríguez, JA 2 , Pérez-Frances,
1 Dept of Microbiology and Cellular Biology, Faculty of
Biology, La Laguna University, La Laguna, Tenerife,
Canary Islands, Spain; 2 Dept of Engineering,
Production and Agricultural Economics. School of
Agricultural Engineering. La Laguna University,
3 Tenerife, Spain; Dept of Plant Biology, Faculty of
Pharmacy, La Laguna University, Tenerife, Spain
A histological study of leaves and stems of two
Leucospermum (Proteaceae) cultivars ('Flame Spike' and
'Tango') has been made from micropropagated (70 days old microplantlets) and field plants, using light and transmission electron microscopy. Both cultivars showed a similar tissue distribution and development. Leaves from in vitro and field plants presented a simple epidermis with isodiametric cells covers by a cuticle.
This layer was much thinner in leaves from in vitro plantlets than from field. Trichomes were unicellular, simples and elongated. Leaves were anphistomatic, and presented reniform and paracytic stomata in both surfaces. The mesophyll presented a parenchyma poorly organized and vascular bundles with a limited development and a poor presence of sclerenchyma in leaves grown in vitro, while field leaves showed an organized palisade and spongy parenchyma with intercellular spaces. Ultrastructural observations showed scattered droplets of phenolic compounds in parenchyma and bundles sheath cells from in vitro plantlets leaves.
The palisade parenchyma cells from field leaves containing phenolic deposits presented as fine-granular and occupy the entire vacuolar volume. Stems transversal section showed a concentric tissue organization both in vitro and field plants. The in vitro stems showed a simple epidermis covered by a thin cuticle compared to the field ones. Cortical parenchyma presented a variable cells layer (8-10 from in vitro plantlets and 6-8 from field plants) and collateral vascular bundles were surrounding a large pith in both. In vitro stems showed poorly developed vascular bundles with a small number of
532 sclenchyma fibers outside them. Ultrastructural observations showed cortical cells containing phenolic deposits presented as scattered droplets in vitro stems, these deposits were lower than in field stems.
'Flamespike' showed scattered droplets, while `Tango´ phenols are presented both scattered droplets and finegranular. 'Flame Spike' and `Tango´ in vitro plantlets showed certain characteristic features due to the microenvironment where they have grown. These features render them vulnerable to the transplantation shock when directly placed in a greenhouse or field. This work gives us guidelines for developing an optimal acclimatization protocol that increase survival rates in greenhouse and field.
P0612 – ePoster
1
Xplo: a software for plant architecture exploration
Griffon, S 1 , Barczi, J-F 1 , de Coligny, F 2
CIRAD - UMR AMAP, Montpellier, France; 2 INRA -
UMR AMAP, France
Improving our knowledge on plant architecture and development will help to answer remaining questions in ecology, agronomy and forestry, i.e. regarding plant acclimation and adaptation to changing environments or optimization of plant products. For this purpose, computational plants are becoming more and more popular and a lot of effort is made to build structural and functional models at the organ level in order to simulate plant growth and structure [1]. In this context, scientists need software to measure, analyse and model plant architectures. The Xplo project has been developed in
UMR AMAP since 2008 aiming at edition, visualization, exploration and simulation of plant architecture in a userfriendly manner. The software gives interactive tools to handle the plant structure topology (i.e. organ addition/ deletion), geometry (i.e. 3D selection, edition, rotation) and dynamic (i.e. time line, scenarios). It has been used for various types of plant architecture studies (i.e
Sunflower [2], Fir Sapling[3], Cecropia, Palm trees [4]).
Xplo is fully compatible with the multi-scale tree graphs
(MTGs) data structure, which is commonly used to represent plant topology [5]. Users can explore this data structure to find or to improve hypotheses on plant development. Specific data can be extracted with combination of criteria and be visualised in tables and graphs. Simple analysies functions can be launched or data can be exported to external tools, i.e. AMAPMod, R or any other statistical computing environment, for more specific analyses. Once the model is designed, modellers can integrate their own plant simulation model to the platform and get benefit of its functionalities to test their hypothesis. Different scenarios can be computed for a growth model by interactively modifying model parameters or plant structure (i.e. by pruning) at defined time steps. A plant geometry builder is integrated to
Xplo. It allows a 3D plant mockup to be computed according to available geometrical data in the tree graph.
If geometrical information is missing, standard default algorithms are provided to compute the missing data and to make 3D rendering available. Xplo is an expandable software built around the stable Capsis kernel [6].
Independent simulation models are integrated in Xplo under the form of separated modules, and various tools

can be plugged at any time using flexible extensions.
This generic software can be run either in an interactive context with a multi-language graphical user interface or in script mode (Groovy/Python). Xplo is a free opensource software (LGPL) and is available on almost every
OS.
P0613 – ePoster
Ecological trends in wood anatomy: is it possible to define patterns in species of Atlantic rain forest
1 domain?
Barros, C 1 , Callado, C 2
Instituto de Pesquisas Jardim Botanico do Rio De
Janeiro, Brazil;
Janeiro, Brazill;
2
, Da Cunha, M 3 , Lima, HRP 3
Universidade do Estado do Rio de
3 Universidade Estadual do Norte
Fluminense
The Atlantic rain forest (ARF), one of the 25 biodiversity hotspots for conservation priority, is a mosaic of different physiognomies. The biome that once covered most of the
Brazilian east coast is now reduced to 7%. Despite its impressive biodiversity, little is known about the wood anatomy of ARF species and understand the variation of the wood anatomy is a key role to evaluating environmental and biotic influences in ecosystems. To gain more insight into the ecological trends in wood anatomy, qualitative and quantitative wood anatomy parameters of species from three remnant in ARF domain were compared (256 tree specimens, belonging to 82 species and 25 families) and the results were compared with literature data of 187 species from 9 different forest remnants around the world: one in Amazon forest
(Fedalto et al. 1989), two in Mexico (Barajas-Morales
1985), one in Venezuela (Lindorf 1994) and five in
Southwestern Australia (Carlquist 1977), to answer the following questions: (1) can wood anatomy parameters be used to characterize different physiognomies of the
ARF?; (2) can an ecological trend for the remnants of the
ARF be identified? and, if so, (3) is it comparable to results from other biomes? The samples were collected in
Rio de Janeiro State in two Montane forest patch at the
Parque Nacional do Itatiaia (PNI) (22º15’– 22º30’S and
44º30’– 44º45’W) and Reserva Ecológica de Macaé de
Cima (REMC) (22º21’– 22º28’S; 42º27’– 42º35’W) and one Lowland forest sampled in Reserva Biológica de
Poço das Antas (RBPA) (220 30’– 220 33’S; 420 15’–
420 19’W). Our results showed that the ARF species have privileged the efficiency of the conduction investing in low frequency (average of 20) of vessels wider than 80
µm with simple perforation plates. The principal component analysis separates the three remnants in different groups, which allows us to detect features for each physiognomy: the Montane forests are characterized by vessel elements longer than 500 µm and narrower than 100 µm and septate fibres (52% PNI and 71%
REMC). The PNI species are separated from REMC by the incidence of distinct growth rings boundaries (86% in
PNI and 57% in REMC). The RBPA species presented distinct growth rings boundaries (82%), vessel frequency lesser than 10 and vessels wider than 100 µm. The comparison of ARF remnants and the literature data, although applied to only a few vessel element characters, demonstrate in a statistical approach that the wood anatomical features can characterize different forest remnants. The more xenomorphic remnants from
Australia remain grouped. The dry forest in Venezuela and the tropical deciduous forest from Mexico integrate another group, as well as the tropical Lowland forests from Mexico, Amazon and ARF (RBPA) and the
Montane forests of ARF.
P0614 – ePoser
Leaf anatomy of invasive and non-invasive climbers: does it correspond with ecophysiological
1 performance?
Boyne, RL 1 , Osunkoya, OO 1 , Scharaschkin, T 1
Queensland University of Technology, Australia
An aspect of invasion biology is the identification of common traits of invasive species that may contribute to their invasiveness. A previous study that compared resource-use efficiency and phenotypic plasticity of invasive and non-invasive climbing plants in Southeastern Queensland under different light conditions indicated that invasive species tend to have a more efficient use of light and carbon compared with noninvasive counterparts, while non-invasive species have a greater water-use efficiency. Furthermore, the invasive species were shown to have a lower leaf construction cost and greater plasticity in such structural traits as leaf thickness and density. This set of characteristics may contribute to the fitness of the invasive species. The present study compares leaf anatomical and morphological traits associated with relevant ecophysiological traits of the same species grown under different light conditions. These anatomical traits were examined primarily by light microscopy of stained leaf sections and impressions of the epidermis. Specifically, it was hypothesised that invasive species have anatomical traits consistent with a high light-use efficiency (e.g. fewer trichomes), low water-use efficiency (e.g. thinner epidermis) and low construction cost (e.g. less lignin) than non-invasive counterparts, and that they have a greater ability to adjust their anatomy to different light levels. A possible exception to this generalisation may be given to one non-invasive species (Parsonsia straminea), which has an ecological performance closer to that of invasive ones.
P0615 – ePoster
Ultrastructure of the cells taking part in oogenesis of
1 the fern
Osmunda japonica
Cao, J 1 , Dai, X 1 , Wang, Q 1
College of Life and Environmental Sciences, Shanghai
Normal University, China
The development of the egg and canal cells in the fern
Osmunda japonica Thunb. was examined by transmission electron microscopy. The present studies show that the mature egg possesses no typical egg envelope and fertilization pore, which are considered to be the prominent features of the mature egg in leptosporangiate ferns. An extra wall, possibly callose in nature, forms in the canal cells during oogenesis of
Osmunda japonica , but this special wall is not present in
533

other leptosporangiat ferns. A separation cavity forms above the egg cell possibly by exocytosis of the egg. The egg nucleus retains spherical shape and no nuclear evaginations occur during oogenesis. Amyloplasts in the egg cytoplasm are numerous and conspicuous. Most of them surround and lie closely to the nucleus. The egg and the canal cells, isolated by the extra wall in most stages, develop independently, which differ greatly from the leptosporangiate ferns. The cytological features of the egg and the canal cells of O. japonica in oogenesis are most probably primitive in the plant sexual evolution.
The present results suggest that Osmundaceae should be separated from leptosporangiopsida.
P0616 – ePoster
Fruit anatomy and morphology of
Tabernaemontaneae (Apocynaceae, Rauvolfioideae) with taxonomic notes
Vilalba-Ferreira, CV 1 , Simões, AO
1
1 , Carmello-
Guerreiro, SM
1 Plant Biology Dpartment, Institute of Biology,
University of Campinas-UNICAMP, Campinas, Brasil
Rauvolfioideae comprises 10 tribes and about 850 species that show an unparalleled variation in fruit and seeds features within Apocynaceae. From the tribes currently recognized in Rauvolfioideae,
Tabernaemontaneae is an ideal group for evolutionary studies. A significant morphological variation of reproductive traits is coupled with a deep understanding of phylogenetic relationships among its taxa. Two subtribes, Ambelaniineae and Tabernaemontaniineae, have been recognized in Tabernaemontaneae, both characterized by fruit and seed features. Ambelaniineae has baccate fruits and seeds without arils, whereas
Tabernaemontaniineae has follicular fruits, more rarely baccate, and seeds partially to totally covered by arils. In the present study, the morphology and anatomy of fruits from four species of Ambelaniineae ( Ambelania duckei,
Macoubea sprucei, Rhigospira quadrangularis,
Spongiosperma macrophyllum ) and one of
Tabernaemontaniineae ( Tabernaemontana sananho ) were analyzed, aiming to identify diagnostic characters with potential evolutionary and taxonomic value. In T. sananho , the epicarp is unisseriate and formed by cells with thick walls, has a thin cuticle, and a conspicuous hypoderm. The mesocarp has sclereids, laticifers, scattered vascular bundles and well-developed aerenchyma. The endocarp is unisseriate and formed by cells with thick walls. In the studied species of
Ambelaniineae, the epicarp is unisseriate and formed by cells with thin walls, has a thick cuticle and no hypoderm. The mesocarp has sclereids, laticifers, scattered vascular bundles, several idioblasts containing phenols, and no aerenchyma. The internal cells of the mesocarp form inward protusions that partially cover the seeds, a feature not observed in T. sananho . The endocarp is unisseriate and formed by cells with thin walls. A flange-shaped cuticle was observed on the epicarp surface in A. duckei, R. quadrangularis and S. macrophyllum , but not in M. sprucei . The sister relationship of Macoubea to the other genera of
Ambelaniineae suggest that a flange-shaped cuticle might have evolved once after the divergence of Macoubea and
534 thus not represent a sinapomorphy for the subtribe. The baccate fruits of T. sananho (the only known neotropical species of the genus with indehiscent fruits), though superficially similar to the fruits of Ambelaniineae, share a number of anatomical features with the follicular fruits of Tabernaemontaniineae species. The most important of these shared features is the presence of a dehiscence line that is vestigial and non-functional in T. sananho . This suggests that the fruit of that species has evolved from an ancestor with follicular fruits.
P0618 – ePoster
New observations on anther and pollen development in
Hypolytrum and
Mapania
(Mapanioideae,
1
Cyperaceae)
Coan, AI 1 , Alves, M 2
Departamento de Botanica, Instituto de Biociencias de
Rio Claro, Universidade Estadual Paulista (UNESP),
Rio Claro, Brazil; 2 Departamento de Botânica,
Universidade Federal de Pernambuco (UFPE), Recife,
Brazil
Anther and pollen development in Hypolytrum schraderianum and Mapania pycnostachya was studied, with special reference to the early stages of microsporogenesis, in order to contribute to the palynological characterization of Hypolytreae
(Mapaniodeae). Inflorescences were collected in northeastern and southeastern Brazil and fixed for light microscopy. A remarkable feature of H. schraderianum is the arrangement of the microsporocytes within the anther locule, corresponding to a single cylinder of cells.
Such peripheral arrangement is maintained up to the initial stages of microgametogenesis, by the time the three non-functional nuclei degenerate. At this stage the pseudomonads become centrally arranged within the anther locule. Phenolic idioblasts occur within the connective cells of H. schraderianum . In addition, early stages of microsporogenesis in M. pycnostachya provide evidence of typical pseudomonad development. In this species, in grains located at the edge of the anther locule, the functional nucleus is positioned close to the wall adjacent to the secretory tapetum; the three nonfunctional nuclei are located along the opposite wall.
This observation in Mapania , as previously reported for
Hypolytrum , reinforces the occurrence of pseudomonads in Hypolytreae and reveals the uniformity of pollen development within both genera in Mapanioideae.
P0619 – Poster
Macadamia
development: branching and flowering
Conway, J 1 , Hanan, J 1 , Stephenson, R 2
1 University of Queensland, Australia;
Industries Queensland, Australia
2 Primary
Macadamia ( Macadamia tetraphylla and M. integrifolia ) axillary buds can form either branches or inflorescences.
Current macadamia pruning methods result in outgrowth of numerous branches producing thick canopy walls.
These shade canopy interiors and increase axillary bud dormancy in the centre of the tree. Greater understanding

of macadamia axillary bud behaviour may assist orchard managers with pruning methods that sustainably maximise nut yields. Initial results from this project show axillary bud position along the growth unit affects the probability of floral induction. It has also been shown that floral induction and inflorescence emergence are promoted by different night temperatures. New work is investigating effects of apical vigour on inflorescence production, and the time of year of floral induction. The interactions of these natural behaviours with different pruning cut locations will then be examined.
P0620 – ePoster
PAP signals from the chloroplast regulate exoribonucleases, gene silencing and stress responses in
Arabidopsis
Crisp, P 1 , Estavillo, G 1
Giraud, E 3 , Hell, R 2
Heidelberg, Germany;
, Pornsiriwong, W 1
, Whelan, J 3 , Searle, I 1
, Wirtz, M 2
, Pogson, B 1
1 Australian National University, Australia; 2
,
University of
3 University of Western Australia,
Australia
Chloroplasts regulate nuclear gene expression during plant development and in response to environmental stresses via retrograde signalling pathways. Here we report a novel chloroplast-nucleus signal that regulates
RNA metabolism and gene silencing in the nucleus. In a genetic screen to identify components of the chloroplast stress signalling pathways we isolated the Arabidopsis mutant altered APX2 expression 8 (alx8) [1]. The alx8 mutant exhibits constitutive up-regulation of stress inducible genes, including APX2 and ELIP2, and drought tolerance. alx8 is a mutation in the SAL1 phosphatase [2], which we have shown to be localised to the chloroplast. Loss of chloroplastic SAL1 leads to a 10 fold elevation of its substrate, 3-phosphoadenosine 5phosphate (PAP), which is an inhibitor of 5’-3’ exoribonucleases (XRNs) in yeast [3]. Building on an earlier report linking SAL1 to the activity of the exoribonucleases in plants [4], we show that stressinducible genes elevated in the SAL1 knockouts – alx8 and fiery1 – are similarly up-regulated in the xrn2 xrn3 double mutant. Global transcriptsome analysis of alx8 and xrn2 xrn3 reveals that both SAL1 and the XRNs modulate the expression of a very similar subset of genes
(56%). The significant overlap between the alx8 and xrn2 xrn3 transcriptomes strongly supports the model that
SAL1 and the nuclear XRN2 and XRN3 function in the same pathway regulating the expression of large number of genes. Furthermore, we show that the xrn mutants are more drought tolerant that wild-type plants. Thus, these results implicate exoribonucleases in stress response and gene regulation, and establish a compelling link between chloroplast signalling, RNA metabolism and gene expression. [1] Rossel et al. (2006).[2] Wilson, Estavillo et al. (2009). [3] Dichtl et al. (1997). [4] Gy et al. (2007).
Research supported by the ARC Centre for Plant Energy
Biology (CE0561495); and GRDC Scholarship
(GRS184).
P0621 – ePoster
Cell totipotency and life program cascade: a botanical perspective
1
Cui, K-M 1
College of Life Sciences, Peking University, Beijing, PR
China
Recently cell reprogramming has become a hot research and some exciting results have been obtained in exploiting totipotency of differentiated animal cells.
However, the totipotency nature of tissue cells was discovered in 1902 by botanist G. Haberlandt. Research in botany over the past century has shown that life program for multicellular organism developed from reproduction and differentiation of a zygote cell is a cascade consisting of different stages. The developmental potential of a cell is not only limited by the stage of its life program but also influenced by the position information (microenvironment) created over the cascade of development. We have studied tissue and organ regeneration in plants for decades and learned some features of this cascade life program (Cui 1992, 1997,
2007). In this cascade of development cell differentiation and dedifferentiation are two opposite reactions in various stages of life program. A cell may stop at a cascade stage and later resume its further development.
There is also a critical point after which cell differentiation seemed irreversible and programmed cell death (PCD) is the only fate of the cell. However, many cells poised at a differentiation stage before the critical point can be dedifferentiated, and a cell dedifferentiated into any non-terminal differentiation stage can redifferentiate to form an organ or embryoid.
Transdifferentiation can also occur between two types of cells in similar differentiation stage. For example, after removing cambium cells by girdling the tree trunk, cells in surface layers of immature xylem cells (mainly xylem ray cells and occasionally a small number of cambium cells) can dedifferentiate into callus, and then redifferentiated into periderm (IAWA Bul. n.s. 1988).
Cells in some layers of immature xylem cells bellow these surface layers transdifferentiated into phloem cells
(J. Exp. Bot. 2008). And cells in the deep layers of immature xylem cells dedifferentiated into cambium cells
(IAWA Bul. n.s. 1988). When the immature xylem was cultured as some explants in vitro, they dedifferentiated into callus, and then redifferentiated into shoot or root or embryoid (Acta Bot. Sin. 1981). These suggested that the fate of the immature xylem cells left on the girdled trunk surface changed with the location change. Thus, the plastic nature of tissue cells has been shown in botany for many years in supporting Haberlandt's cell totipotency theory. This valuable information should shed light to zoological reprogramming studies which just began to appreciate the 'amazing' totipotency by 'surprisingly' finding some plasticity of isogenetic cells. Inventing a term 'reprogramming' should not increase the confidence of cell engineering when some known features of life program were even forgotten. In contrast, respecting prior knowledge should help 'reprogrammers' to avoid some 'fetal' mistakes. Thus, some 'cross-talks' between botanical and zoological cell biologists may be necessary for yielding some fruitful research results.
P0622 – ePoster
Does ethylene induce lace plant programmed cell death during leaf morphogenesis?
535

Dauphinee, A 1 , Gunawardena, A 1
1 Dalhousie University, Halifax, Nova Scotia, Canada
The lace plant ( Aponogeton madagascariensis ) is a fully submerged aquatic monocot endemic to Madagascar. The lace plant is one of the few known species to produce complex leaf morphology through developmentally regulated programmed cell death (PCD). During leaf morphogenesis, cells are deleted through PCD; beginning in the center, and progressing towards the longitudinal and transverse veins of the leaf, however this cell deletion process stops 4-5 cell layers before the vein borders. The result is perforated leaf morphology with a lace-like pattern, hence the common name. The lace plant has provided a model system for studying plant PCD; however the developmental induction signal of this cellular death process remains unknown. The plant phytohormone ethylene (C2H4) is a gaseous hydrocarbon produced in nearly all tissue types of higher order plants.
Ethylene is a key regulator of many plant development processes including; germination, growth and senescence. Ethylene has also been found to have a role in developmentally regulated PCD induction in cases such as; the deletion of the embryonic suspensor, aerenchyma formation and xylem differentiation. The objective of this study is to determine if ethylene acts as an induction signal for developmentally regulated PCD during lace plant leaf morphogenesis. To determine the role that ethylene plays in lace plant PCD, plants were treated with either the ethylene biosynthesis inhibitor aminoethoxyvinylglycine (AVG), or the ethylene biosynthesis promoter 1-aminocyclopropane-1carboxylic acid (ACC) and compared to controls. Results have shown a significant reduction in ethylene production by lace plants treated with AVG and in contrast, plants treated with ACC had significantly higher ethylene levels. The corresponding leaf morphology of AVG treated plants displayed a significantly lower number of perforations formed. This data suggests that ethylene is involved in lace plant PCD, however more work is being carried out including; investigation of the effects on the number of perforations formed by ACC treated plants, as well as plants treated with a combination of AVG and ACC.
P0624 – ePoster
Morphogenesis, structure and microchemistry of pollinarium in Asclepiadeae
Demarco, D 1
1 Universidade de São Paulo, Brazil
Flowers of Apocynaceae present unusual synorganization between parts and organs which has led to origin of new organs, as the gynostegium. This synorganization of androecium and gynoecium permitted the evolvement of the pollinarium. In the five species of Asclepiadeae studied, the style head possesses palisade secretory epidermis along the lateral surface of its interstaminal areas, responsible for the secretion of the translator.
Translator is composed of corpusculum and two caudicles and its morphogenesis starts at the beginning of the floral development, just after the calyx opening. The translator shape is due to the different activity of the
536 secretory cells which is coordinated spatially and temporally, besides the amount and composition of the secretion, and the ondulated outline of secretory surface.
The secretion shows viscosity aspect since the beginning of its releasing and this is an important characteristic to permit that corpusculum acquires its shape during the secretory process. Corpusculum and caudicles are composed of different chemical substances. Mucilage, fatty acids, phenolic compounds and proteins were detected in the corpusculum, whereas only neutral lipids and mucilage are present in the caudicles. In the anthers, tapetal cells are involved in the secretion of lipids that cover the pollinium, forming a pellicle of sporopollenin.
Besides pellicle, pollinia of Matelea denticulata present particular features, such as pellucid margin and hyaline crest produced by tapetum which joins pollinium to caudicle. Just before anthesis, anthers dehisce and the pollinia from adjacent anthers contact and stick to the caudicles of a translator, forming the pollinarium.
P0626 – ePoster
1
Structural types of frond anatomy of ferns
Derzhavina, N 1
Orel State University, USA
The objective of this work was to detect diversity of frond structural types in homosporous ferns living in various ecological conditions (epiphytes, epilithophytes, helophytes, and hydrophytes), as well to analyze interrelations between structural types in connection with possible directions of structural functional evolution of ferns. Thirty fern species from 9 families (Polypodiaceae,
Aspleniaceae, Woodsiaceae, Adiantaceae,
Dryopteridaceae, Synopteridaceae, Hymenophyllaceae,
Parkeriaceae, Lomariopsidaceae) were studied. The base of the study is the method of analysis of the plant photosynthetic apparatus developed by Mokronossov
(1978) and Mokronossov and Borzenkova (1978). The method includes the complex analysis of plant photosynthetic apparatus at various levels of its organization such as a) fronds, b) mesophyll cells and c) plastid apparatus. Distinguishing of types was based on histological survey of fronds including following parameters: mesophyll type, number of its layers; type of frond in relation to distribution of stomata; type of stomatal apparatus and number of stomata per 1 mm 2 of frond area; occurrence of fibers of sclerenchyma; peculiarities of venation; type of petiole; etc. In addition, peculiarities of photosynthetic apparatus of the studied species are considered and an analysis of the most outstanding functional peculiarities of fronds of these ferns is included. According to our data, anatomical changes of fronds of fern sporophytes in the course of adaptation to various environments are related primarily to structural reorganizations of mesophyll: increase of number of its layers and rise of cell density under xerophytization, increase of cell size under succulentization and, finally, decrease of number of layers to one in shade hygrophytes. In the studied ferns and taking into account literature data, we distinguish following structural types of frond anatomy: hydromorphic, hygromorphic, mesomorphic, subxeromorphic, and subsucculent (cryptic succulent).
To our opinion, following characteristics of fronds are of

highest adaptive value: their area and thickness, mesophyll type, dry weight of an area unit – specific superficial frond density (SSFD), cellular volume, number of cells per an area unit – these are main characters. When attempting to summarize anatomical features being the most significant for ecological diagnosis of frond structural types, in the case of subxerophytes it is primarily apparent dorsoventrality of mesophyll – its differentiation into palisade (clearly identifiable) and spongy tissue, sometimes a hypodermis in addition, and then – high values of dry weight of frond area unit (SSFD) – pycnosis; in the case of subsucculents
– large-celled dorsoventral or homogenous mesophyll and relatively thick blades (distinction of ferns from succulent flowering plants which most important features being high density of mesophyll and its differentiation into several specialized tissues – Gamaley,
Shiyrevdamba 1988) (these data represent an another evidence for suggestion that for a number of functional and structural characters ferns convergently verge to highly specialized xerophytes-heliophytes typical among flowering plants but are not identical to them); in the case of hygrophytes – low values of SSFD and minimal thickness of blades with homogeneous mesophyll; in the case of hydrophytes – besides characters listed for hygrophytes – well-developed system of large intercellular spaces and air canals.
P0627 – ePoster
Seed anatomy of the genus
Alysicarpus
Desv.
Dhabe, A 1
1 Dept of Botany, Dr Babasaheb Ambedkar Marathwada
University, Aurangabad (MS), India
Alysicarpus Desv. is a small genus of family Fabaceae
(Subfamily Papilionoideae, tribe Desmodiae) comprising
42 taxa, 27 species and 15 infraspecific taxa. India harbours 17 species and 9 infraspecific taxa. To know the seed anatomical features seed anatomy of 24 taxa was undertaken. The seeds were collected from different localities of Maharashtra, Gujarat, Andhra Pradesh,
Karnataka, Tamil Nadu etc., the voucher specimens were deposited in ‘BAMU’ Herbarium. The matured seeds were softened in 10% glycerin and 70% ethanol (1:1 v/v) for 2 days and then stored in 70% ethanol. Customary method of dehydration in tertiary butyl alcohol series and embedding in paraffin wax were followed. Microtome sections cut between 8 to 20 µ thickness and stained in safranin-light green combination. Slides were observed under microscope and figures were drawn with the help of a camera lucida. Measurements were taken by calibrating standard micrometry. The keen observations were made on the size, shape of micro sclereids, their lumen, osteosclereids, parenchyma and endosperm layers. It was observed that the lumen of the macrosclereids was peculiar to particular species, its base was of three types flat, convex and concave.
Osteosclereids varies from species to species, they may be compactly arranged without gap (as in A. bupleurifolium and var. gracilis ) or may be loosely arranged with large gap. Their ecorners may be sharp or blunt. Parenchyma may be single to multilayered, small thin walled to large thick walled cells. Endosperm was usually single layered but single to double layered endosperm was reported in A. heyneanus and complete double layered large celled endosperm was found in A. monilifer . Endosperm cells may compactly or loosely arranged, with small or large cytoplasmic contents. It was concluded that, not only species of Alysicarpus but varieties and forms are also specific in seed anatomy.
P0628 – ePoster
Diversification of
ARA6
functions among land plants
Era, A 1 , Ishizaki, K 2 , Kohchi, T 2 , Nakano, A 3
Ueda, T 1
, Ebine, K 1 ,
1 Graduate School of Science, University of Tokyo, Tokyo,
Japan; 2 Graduate School of Biostudies, Kyoto
University, Kyoto, Japan; 3 RIKEN ASI, Saitama, Japan
Single membrane-bound organelles are interconnected by a membrane trafficking system, where Rab GTPases play critical roles as molecular switches. Among Rab
GTPases, RAB5 is known to regulate membrane fusion between early endosomes in animal cells. Arabidopsis thaliana also harbors two RAB5 orthologs, ARA7 and
RHA1 , whose products function in endocytic and vacuolar transport pathways. ARA6 is another RAB5 member with unique structural features in its product, whose orthologs are conserved only in land plants.
Genetic studies demonstrated that ARA6 has totally different functions from conventional RAB5 s. Studies on subcellular localization and function of ARA6 further indicated that ARA6 regulates transport from endosomes to the plasma membrane. To examine whether the functions of conventional and plant-unique ARA6 -like
RAB5 s are conserved among land plants, we are investigating functions of Rab5 members in basal land plants. In this meeting, we present the results of localization and functional analyses of an ARA6 -like
RAB GTPase in an emerging model of the liverwort,
Marchantia polymorpha . We isolated plant-unique RAB5
(Mp ARA6 ) and conventional RAB5 (Mp RAB5 ) from M. polymorpha , and examined their intracellular localization. Both Mp RAB5 and Mp ARA6 were localized on dot-like organelles in thallus cells. These organelles were stained by FM1-43, indicating that both are on endocytic compartments. Intriguingly, the Mp RAB5 positive compartments were tubulated by wortmannin treatment, whereas the Mp ARA6 -positive compartments were insensitive to wortmannin. These results suggested that Mp RAB5 and Mp ARA6 reside on different endocytic organelles. We then examined subcellular localization of
GTP-fixed mutants of these RAB5 proteins. GTP-fixed
RABs are generally thought to accumulate on the target membranes because they could not be detached from membranes by GDI. The GTP-fixed mutant of Mp RAB5 accumulated on vacuolar membranes, while GTP-fixed
Mp ARA6 was on unknown membrane compartments, which were not stained by FM1-43. These results suggested that Mp ARA6 acts in the unique trafficking pathway to the unknown membrane compartments, which also indicated that functions of ARA6 -type RAB
GTPases are diversified among land plants.
537

P0629 – Poster
Removal of cytosolic PPi is a prerequisite for proper resumption of post-germinative development in
Arabidopsis thaliana
Ferjani, A 1 , Muto, Y 2 , Horiguchi, G 3
2
, Maeshima, M 2
Nagoya University,
Japan; Rikkyo University, Tokyo, Japan;
Tokyo, Japan; 5
4
,
1
Tsukaya, H 4,5
Tokyo Gakugei University, Japan;
3 University of
National Institute for Basic Biology,
Okazaki, Japan
Analyses of leaf development in several mutants of
Arabidopsis thaliana revealed that enhanced post-mitotic cell expansion is often triggered by a decrease in cell proliferation activity below a certain threshold. This phenomenon that we named 'compensation' (Tsukaya,
2002) provides clues to understand the way in which the number and size of cells co-operatively interact to determine the final organ size. Among five compensation-exhibiting mutants ( fugu1 fugu5 ) that we have isolated and characterized (Ferjani et al. 2007), here we report our recent findings on the fugu5 mutant. When fugu5 is germinated on inorganic medium (rockwool), cell division is almost completely lost in mesophyll cells of cotyledons, which are oblong in shape and exhibit strong compensation. Interestingly, we found that compensation in fugu5 mutant is cancelled when sucrose is supplied in the growth media. However, the mechanism of sucrose action remained unclear. Cloning of FUGU5 gene revealed that it is AVP1 , which encodes for the vacuolar pyrophosphatase (V-PPase) that has two functions; hydrolysis of cytosolic pyrophosphate (PPi) and vacuolar acidification. So, what is the triggering factor of compensation in fugu5 ? As an approach to answer this question, we introduced the cytosolic inorganic pyrophosphatase 1 ( IPP1 ) gene of
Saccharomyces cerevisiae , under the control of FUGU5 promoter. IPP1 was used because it only hydrolyzes the cytosolic PPi without contributing to vacuolar acidification, thus providing a good tool to analyze
FUGU5 functions independently. Interestingly, our results clearly showed that fugu5 mutant phenotypes were totally recovered after the introduction of
Pro
FUGU5
::IPP1 transgene. Furthermore, the degradation of seed storage proteins, lipid reserves and the peroxisomal
β
-oxidation activity were unaffected in fugu5 . However, as compared to the WT and Pro
FUGU5
::IPP1 transgenic lines, the hypocotyl elongation in fugu5 mutant was severely reduced in the dark and recovered upon addition of sucrose to the growth media.
Together, these findings strongly suggest that gluconeogenesis is compromised in fugu5 mutant due to high cytosolic PPi accumulation. This study provides evidence that PPi hydrolysis, rather than vacuolar acidification, is the major role of FUGU5 in vivo .
Therefore, the removal of cytosolic PPi produced by active cellular metabolism early during germination is a prerequisite for sustaining cell cycling, proper resumption of post-germinative development and to boost seedling growth in Arabidopsis .
538
P0630 – ePoster
Dimorphic chloroplasts in the epidermis of
Podostemaceae aquatic angiosperms and implications for morphological evolution
Fujinami, R 1 , Yoshihama, I 2 , Imaichi, R 1
1 Dept of Chemical and Biological Sciences, Japan
Women's University, Japan; 2 Electron Microscope
Laboratry, Tokyo Medical University, Japan
The Podostemaceae plants are unusual aquatic angiosperms growing in the tropical and subtropical rapids and waterfalls. The plant is subjected to running water continuously during the rainy season. To adapt to such a unique habitat, the Podostemaceae have evolved a unique morphology that deviates extremely from most angiosperms. The root is axial or sometimes foliose looking like the leaf, and functions both as an assimilatory organ and as an attachment to the rock surface. The shoot arising from the root appears to lack stems and is composed only of leaves. There are three subfamilies: Podostemoideae (43 genera, 254 species),
Weddellinoideae (1 genus, 1 species) and Tristichoideae
(6 genera, 15 species). The Podostemoideae with most species exhibit the most unique characteristics, such as loss of the shoot apical meristem (SAM). Consequently, in Podostemoideae, the leaf grows from the base of the previous leaf, not from the SAM. A unique strong chloroplast dimorphism in epidermal cells of assimilatory roots and leaves has been reported previously for Podostemoideae (Fujinami et al. 2011).
This is the first report of chloroplast dimorphism in a single epidermal cell of an angiosperm. Small and large chloroplasts are located separately within each epidermal cell, near the outer and inner walls, respectively, and both are compartmentalized. By contrast, the chloroplasts in cells of the ground tissue are monomorphic with the same size and shape as the large chloroplasts in the epidermis. The small chloroplasts have normal granal ultrastructure, but very few starch grains, while the large chloroplasts have well-developed starch grains. This chloroplast dimorphism is shown in the meristematic region of the root tip, which is exposed to sunlight, but not in the very young leaf primordia seated deeply in some growing leaf primordia. This suggests that the chloroplast dimorphism is an adaptive characteristic, and the small chloroplasts along the outer tangential wall probably function in CO use HCO
3
as the CO
2
2
; uptake for photosynthesis from the torrential water to supply energy to the HCO
source via the HCO
3
-
3
pump, because most submerged freshwater angiosperms
pump.
Podostemoideae have a much wider range of morphology and size than Tristichoideae, the most basally diverged clade. We do not know how Podostemoideae have evolved such a varied morphology while all living in a similarly uniform environment (running water).
Tristichoideae do not show chloroplast dimorphism, suggesting that this feature was acquired when the
Podostemoideae clade diverged from the rest of the
Podostemaceae. Acquisition of small chloroplasts in epidermal cells, which may facilitate CO uptake, might
2 have allowed the evolution of the extreme diversity of morphologies in Podostemoideae.

P0631 – Poster
The formation of flooding-induced vascular cavities in
Pisum sativum
primary roots occurs via programmed cell death
Sarkar, P 1 , Sreekanta, S 2 , Niki, T 3 , Gladish, DK 4
1 Energy Biosciences Institute, University of California-
Berkeley, USA; 2 Dept of Plant Biology, University of
3 Minnesota-Twin Cities, St Paul, USA; Dept of Cell
Technology, Takushoku University, Hachioji, Japan;
4 Dept of Botany, Miami University, Hamilton, USA
Many organisms sacrifice specific cells in their body by programmed cell death (PCD) in order to survive biotic and abiotic stresses. PCD follows controlled pathways and affects only specific cells or tissues without spreading randomly to neighboring cells. Several different types of PCD have been described based on the cytological events that accompany the cell death. Primary roots of garden pea ( Pisum sativum ) respond to flooding at warm temperature (25° C) by forming a longitudinal cavity in the center of their vascular cylinders (VC).
Several lines of evidence strongly suggest that vascular cavities form via PCD. Pea seedlings were grown under sterile conditions in 2 L beakers filled with moistened vermiculite at 25°C for 4 d, then flooded with deionized water or caspase inhibitor solutions for various times up to 24 h. Controls were unflooded. Samples were taken a various times during the test period.
Ultrastructural features of relevant cells were examined by transmission electron microscopy. Fluorescence microscope techniques, including TUNEL and cytochrome c immunolabeling, were used to visualize
DNA fragmentation and mobilization of cytochrome c from mitochondria in situ. Mitochondria were isolated from cytosol and both were evaluated for changes in cytochrome c quantities over time using Western blotting. DNA was extracted and pulse-field gel electrophoresis was used to detect DNA fragmentation.
Total protein extracts were assayed for YVADase and
DEVDase activity by fluorometric spectrophotometry.
Systematic DNA fragmentation, a hallmark character of most PCD pathways, was detected in the vascular cavityforming pea primary roots. DNA fragmentation occurred rapidly, within 3 h after flooding. The DNA fragments produced were about 20-30 kb, corresponding to the approximate size of chromatin loops in pea chromosomes. Release of cytochrome c from mitochondria into cytosol, a characteristic feature of a common PCD pathway, was detected in the cavityforming roots after 2 h of flooding, earlier than visualization of DNA fragmentation by TUNEL. DNA fragmentation and cytochrome c release remained confined to the parenchymatous cells in center of the
VCs. YVADase and DEVDase activity increased significantly within 6 h of flooding, but declined to control levels by 24 h under flooding. Inhibitors specific to these enzymes minimized vascular cavity formation.
Affected cells in VCs displayed morphological distortion of nuclei and other organelles followed by rupture of the tonoplast and extreme degradative thinning of cell walls.
This ultimately led to total collapse of the cells and the formation of a lengthy longitudinal cavity in the center of the VC. Outer VC cells and cortical cells maintained cellular integrity and had signs of normal activity.
The results of these studies suggest that vascular cavity formation in pea primary roots involves a cascade of events including mitochondrial release of cytochrome c, activation of caspase-like enzymes, and cytoplasmic and nuclear degradation consistent with autophagic PCD.
P0632 – ePoster
TDIF peptide signaling regulates vascular stem cell proliferation via the WOX4 Homeobox gene in
Arabidopsis
Hirakawa, Y 1 , Kondo, Y 1 , Fukuda, H 1
1 University of Tokyo, Bunkyo-ku, Japan
The indeterminate nature of plant growth and development depends on stem cells that are localized in the specific proliferative tissues called meristems. The vascular meristem is one of such indeterminate meristems and is essential for secondary growth of plant body. Previously we showed that cell–cell signaling, mediated by a ligand–receptor pair made of a CLE
(CLV3/ESR-related) family peptide, TDIF (tracheary element differentiation inhibitory factor), and an LRR–
RLK (leucine-rich repeat receptor-like kinase) family of membrane kinase, TDR/PXY (TDIF RECEPTOR/
PHLOEM INTERCALATED WITH XYLEM), regulates proliferation and differentiation of undifferentiated procambial/cambial cells in the Arabidopsis vascular meristem. Exogenous application of TDIF promotes proliferation of procambial cells and suppresses their xylem differentiation in planta. However, the molecular mechanism of the intracellular pathway downstream of
TDIF–TDR, which regulates both proliferation and xylem-differentiation of the procambial cells, is largely unknown. To identify components of this pathway, we searched for genes whose expression levels were altered by TDIF. Of them, a WUSCHEL-related HOMEOBOX gene, WOX4, was rapidly up-regulated in response to
TDIF application in a TDR-dependent manner. WOX4 is expressed preferentially in the procambium and cambium. Genetic analyses showed that WOX4 is required for promoting the proliferation of procambial/cambial cells but not for repressing their differentiation into xylem cells in response to the TDIF signal. Thus, at least two intracellular signaling pathways that diverge after TDIF recognition by TDR might regulate independently the behavior of procambial cells.
Detailed observations in loss-of-function mutants revealed that TDIF–TDR–WOX4 signaling plays a crucial role in the maintenance of the vascular meristem organization during secondary growth and stem cell maintenance in this meristem.
P0633 – ePoster
Analysis of DNA damage response in male gametes of
Cyrtanthus mackenii
during pollen tube growth
Hirano, T 1 , Takagi, K 2 , Hoshino, Y 3 , Abe, T 1
1 RIKEN Nishina Center, Japan; 2 Wakasa Wan Energy
Research Center, Japan; 3 Hokkaido University, Japan
539

Pollen grains are exposed to environmental mutagenic agents such as UV light and ionizing radiation. During the processes of pollination and pollen tube growth as well as double fertilization, DNA damages would be induced in the pollen grains including the male gametes.
It is assumed that the male gametes are needed to repair the DNA damages to transmit the accurate genome information to the next generation. The DNA damage response in the male gamete, however, is still unclear. In the present study, to clarify the DNA damage response, we examined to expose heavy-ion beam which can induce DNA double strand break (DSB) to the pollen grains of Cyrtanthus mackenii , and analyzed the male gamete behavior during pollen tube growth by using in vitro culture system. At first, we investigated germination rate of the pollen grains after irradiation of
C-ion beam at the dose range from 10 to 80 Gy. As a result, the rates showed no decrease compared to the nonirradiated pollen grains. Since this plant species forms bicellular pollen, we focused on the male gamete behavior such as sperm formation during the pollen tube growth. The sperm formation rates were decreased with the dose increase: when 40 and 80 Gy were applied, the sperm formation rates were 55% and 23%, respectively.
By the irradiations at 40 and 80 Gy, the generative cells were largely stopped at metaphase in the pollen mitosis II
(PM II). Therefore, we supposed that DNA damage of the generative cells irradiated with high dose was not repaired completely during pollen tube growth. To elucidate this assumption, we attempted to detect the phosphorylated histone H2AX (
γ
H2AX) as an indicator for DSB in the male gametes during the PM II. In the male gametes derived from the pollen grains irradiated with 40 and 80 Gy,
γ
H2AX focus was not detected in the sperm cells, anaphase and telophase cells. On the other hand, the generative cells and the metaphase cells formed
γ
H2AX foci. These results suggest that one of the reasons for the increase of the metaphase cells in the generative cells was due to the DSBs remaining unrepaired, and spindle check point might be working rather than G2/M arrest. We now attempt to analyze the DNA repair process in the male gametes.
P0634 – ePoster
Expression and regulation of anther-specific genes in the tapetum and microspore of
Lilium longiflorum
Hsu, S-W 1 , Wang, C-S 1
1 Graduate Institute of Biotechnology, National Chung
Hsing University, Taichung
Four stage-specific genes (LLA-67, LLA-89, LLA-115, and LLA-142) have been isolated from a subtractive cDNA library constructed from developing anthers of lily
( Lilium longiflorum ). The proteins encoded by three of these genes (LLA-67, LLA-89, and LLA-115) have a strong hydrophobic region at the N-terminus, indicating the presence of a signal peptide. Sequence alignment revealed that the deduced LLA-67 is a new type of small cysteine-rich protein whose sequence exhibits four consecutive CX3CX6_10 repeats that could form signalreceiving finger motifs, and the deduced LLA-115 protein shows significant similarities to a rice unknown protein, and putative cell wall proteins of Medicago truncatula and Arabidopsis . In addition, the protein
540 encoded by LLA-142 is novel, while the protein encoded by LLA-89 is identical to a reported LIM4 protein with unknown function. Total RNA blot analysis indicated that the transcripts of these genes were anther specific and differentially detected at the phase of microspore development. In situ hybridization with antisense riboprobes of the four genes in the anther showed strong signals localized to the tapetal layer of the anther wall.
The LLA-67, LLA-89, and LLA-142 mRNAs were also detected in the microspore at the phase of microspore development but the LLA-115 mRNA was not.
Furthermore, the LLA-89 and LLA-115 genes can be exogenously induced by gibberellin (GA), whereas the
LLA-67 and LLA-142 genes can not be induced. Studies with the GA biosynthesis inhibitor uniconazole and an inhibitor of ethylene activity, 2,5-norbornadien (NBD), revealed that the LLA-67 and LLA-115 genes were negatively regulated by ethylene and a cross-talk between GA and ethylene was involved in the regulation of the two genes occurring in young anthers.
P0635 – ePoster
Fertility restorer gene Rf2 for lead rice type cytoplasmic male sterility of rice encodes mitochondrial glycine-rich protein
Itabashi, E 1 , Iwata, N 1 , Fujii, S 1 , Kazama, T 1 , Toriyama,
1
K 1
Graduate School Agri. Sci., Tohoku Univ., Sendai,
Japan
Cytoplasmic male sterility (CMS) is a maternally inherited trait that results in the inability to produce fertile pollen and is often associated with an unusual open reading frame found in mitochondrial genomes. The fertility of CMS plant is restored by the nuclear-encoded fertility restorer gene (Rf). Although several Rf genes, many of which encode a mitochondrial Pentatricopeptide repeat (PPR) protein, have been identified so far, the mechanisms of regulation of mitochondrial gene by the
Rf gene remain largely unknown. Molecular characterization of novel type Rf gene is important to elucidate this process. Lead Rice (LD)-type CMS rice possesses a cytoplasm of the indica variety ‘Lead Rice’ and its pollen fertility is restored by a single nuclear gene
Rf2 in a gametophytic manner. However, the character of
Rf2 has not been molecularly defined yet. In this study, we carried out map-based cloning of Rf2 and proved it encodes a protein consisting of 152 amino acids with a glycine-rich domain in its C terminus. There was only a single nonsynonymous nucleotide polymorphism in the orf resulting in a subsitution of isoleucine in the fertility restoring allele (Rf2) by threonin in the non-restoring allele (rf2) at the 78th amino acid residue. The expression analysis showed mRNA of Rf2 (rf2) was strongly detected in anthers at the bicellular and tricellular pollen stages in the both restoring allele and non-restoring allele. The transiently expressed RF2-GFP and rf2-GFP fusion proteins were shown to be targeted to mitochondria. The above results suggested a single polymorphism in amino acid in rf2 did not affect its localization to mitochondria and was a cause of the function deficiency in mitochondria. Unlike other RF factors, which are PPR protein considered to bind to the target mRNA in their CMS/Rf systems, RF2 did not

seem to possess putative RNA-binding domain. This suggests Rf2 is a novel type Rf gene and the data from this study provide new insights into the mechanism for restoring fertility in CMS.
P0637 – ePoster
Pre and post pollination changes in amino acid and mineral compositions of anther and stigma in
Solanum surattense
Burm.f.
Jain, B 1
1 MG Science Institute, India
In the present investigation an attempt was made to understand the composition of free amino acids and minerals present in anther and stigma before, during and after pollination stages in Solanum surattense Burn.f. employing the techniques of paper chromatography and atomic absorption spectrometry. Chromatographic analysis of anther and stigma revealed that before pollination, both these organs contained only 11 types of amino acids. But during and after pollination, the anther showed the presence of 15 different types of amino acids. This indicates that during and after pollination four new types of amino acids viz. arginine, aspartic acid, ornithine and valine were synthesized in the anther.
Similarly stigma also revealed the presence of 16 different types of amino acids during and after pollination. This indicates the synthesis of five new types of amino acids viz. aspartic acid, lysine, ornithine, proline and valine in the stigma as a result of act of pollination. Atomic absorption spectrophotometric analysis of anther and stigma has revealed the presence of three major elements i.e. K, Ca, and Mg and nine minor elements i.e. Na, Cu, Mn, Ni, Zn, Pb, Fe, Cd and
Cr in the stigma before, during and after pollination stages. A considerable decrease in the amount of K and
Mg and an increase in Ca was observed in the stigma during post pollination stages. In the minor elements Ni,
Zn and Fe showed considerable decrease, while Cu, Mn,
Pb, Cd and Cr showed an increase in the amount in the stigma after pollination. The present paper describes not only the role of amino acids, but also of the major and minor mineral elements in pollen – pistil interaction.
P0642 – ePoster
Vegetative and reproductive growth of
Arabidopsis under microgravity
Karahara, I 1 , Suto, T
Yano, S 4 , Tanigaki, F
Kasahara, H 7
1
4
, Yamaguchi, T
, Shimazu, T 5
, Kamisaka, S 1
2 , Tamaoki, D 3 ,
, Kasahara, H 6 ,
1 Dept of Biology, Graduate School of Science and
Engineering, University of Toyama, Japan; 2 Dept of
Biology, Faculty of Science, University of Toyama,
Japan; 3 Dept of Life Science, Graduate School of Life
Science, University of Hyogo, Japan;
Exploration Agency, Japan;
Japan; 6
5
4 Japan Aerospace
Japan Space Forum,
Japan Manned Space System Ltd, Japan; 7 Dept of Bioscience and Technology, School of Biological
Science and Engineering, Tokai University
Gravity is considered to be one of the most important environmental factors for growth and development of plants throughout their life cycle. We have designed an experiment, which is called Space Seed, to investigate the effects of microgravity on the seed to seed life cycle of plants. We have carried out this experiment using a newly developed apparatus, which is called the Plant
Experiment Unit (PEU) and installed in the Cell Biology
Experiment Facility (CBEF) onboard International Space
Station (ISS) KIBO module. The CBEF is equipped with a turntable generating artificial gravity to perform 1-G control experiment as well as micro-G experiment on board. Arabidopsis seeds sown on dry rockwool in 8
PEUs were transported from Kennedy Space Center to the ISS by Space Shuttle Discovery in STS-128 (17A) mission. This experiment was started on September 10,
2009 and terminated on November 11, 2009. Arabidopsis seeds successfully germinated, and the plants passed through both vegetative and reproductive processes, such as formation of rosette leaves, bolting of inflorescence stems, flowering, formation of siliques and seeds.
Morphometric analyses of the plants on board were performed and obtained parameters were compared among micro-G plants, 1-G control, and the ground control.
P0643 – ePoster
Development of the Casparian strip is delayed by blue light in pea stems
1
Karahara, I 1 , Takaya, E
Weller, J 3 , Reid, J
1 , Fujibayashi, S
3 , Sugai, M 1
1 , Inoue, H 2 ,
Dept of Biology, Graduate School of Science and
Engineering, University of Toyama, Japan; 2 Dept of
Environmental Biology and Chemistry, Graduate School of Science and Engineering, University of Toyama,
Japan; 3 School of Plant Science, University of Tasmania,
Australia
To understand the regulatory mechanisms involved in tissue development by light, the kinetics of the regulation of Casparian strip development in garden pea stems was studied. We found that short-term irradiation with white light delayed the development of the Casparian strip, and used this delay to assess the quantitative effect of light on strip development. We examined the effect of the duration and fluence rates of white light treatment on strip development and observed a significant relationship between fluence and the delay in strip development indicating that the Bunsen-Roscoe law of reciprocity holds for this response. The effect of white light was not inhibited in the presence of a photosynthetic inhibitor,
DCMU, or a carotenoid biosynthesis inhibitor,
Norflurazon, indicating that the delay in the strip development by light is a photomorphogenetic response rather than a subsidiary effect mediated by photosynthetic activity. An action spectrum for the response displayed a major peak in the blue light region, suggesting a dominant role for blue light receptors. A minor peak in the red light region also suggested the possible involvement of phytochromes. Although phytochromes are known to contribute to blue-light responses, phytochrome-deficient mutants showed a normal delay of Casparian strip development in response to blue light, indicating that the response is not mediated
541

by phytochrome and suggesting a role for one or more specific blue light receptors.
P0644 – ePoster
Implications of tree architecture and carbohydrate allocation in macadamia tree development
Karimaei, S 1 , Hanan, J 2
1 School of Biological Science, University of Queensland,
Australia; 2 Centre For Biological Information
Technology (CBIT), University of Queensland, Australia
High density orchards of macadamia have caused problems with canopy management practices, obstructing machinery access and reducing spray efficiency. To discover possible alternatives rather than hedging to resolve these problems, more knowledge about the balance between carbon allocation and plant architectural development are needed. The aim of this research study is to understand the architecture of the macadamia tree and the relationship between tree architecture and carbohydrate allocation during vegetative growth.
Computer modelling alongside physiological experiments are used to further our knowledge in this area. Three experiments are designed for this study; an architectural experiment to understand macadamia tree architectural development/morphogenesis and testing possible dwarfing rootstocks effect on it, a modelling experiment to find the relationship between number of leaves and growth rate of a new flush by using L-system and an experiment to find the relationship between carbohydrate allocation and photosynthesis rate. As a part of this study architectural features of three macadamia genotypes (M. jansenii, the hybrid Beaumont which performs well as a rootstock and the Hidden
Valley variety, A4) were measured during early tree development. M. jansenii showed some dwarfing characteristics: smaller leaves, wide branching angle and different growth pattern.
P0645 – ePoster
Body plan of Hypericaceae and implications for the evolution of Podostemaceae
Koi, S 1 , Kato, M 2 , Nakajima, K 1
1
2
Nara Institute of Science and Technology, Japan;
National Museum of Nature and Science, Japan
Podostemaceae is an aquatic angiosperm family that grows on rocks under running water. The Podostemaceae have evolved a unique body plan. Seedlings of most species have an adventitious root on the lateral side of the hypocotyl instead of the radicle at the bottom. The adventitious roots prostrate on rock surfaces and produce adventitious shoots on their dorsal or lateral sides. The uniqueness of body plan of Podostemaceae is an example of saltational evolution caused by heterotopic modification in development. To reveal how the body plan of Podostemaceae evolved from a typical body plan of angiosperm, we investigated the development of
Hypericum perforatum of Hypericaceae, which is sister to Podostemaceae. Our studies revealed that (1) the seedling of H. perforatum has a plumule and a radicle at
542 the top and the bottom of the hypocotyl, respectively, (2) the seedling produces an adventitious root from the pericycle and endodermis adjacent to xylem of the hypocotyl, (3) some of the roots grow horizontally under the ground and produce adventitious shoots on their lateral sides, (4) excision of the epicotyl from the seedling induces adventitious shoot formation from the pericycle adjacent to xylem of the radicle. The results indicate that the body plan of H. perforatum is similar to that of Terniopsis , which is phylogenetically a basal genus in Podostemaceae. In the seedling of Terniopsis , a plumule and, exceptionally, a radicle occur, and an adventitious root arises near the vascular strand of the hypocotyl (Kita and Kato 2005). This similarity suggests that the seedling of the terrestrial common ancestor of the two families probably had the same body plan as
Podostemaceae and supports Kita and Kato (2005)’s hypothesis that loss of radicle occurred within
Podostemaceae. On the other hand, it is uncertain whether the ability of root-borne shoot formation of
Podostemaceae is comparable to that of Hypericaceae, because the root-borne shoots arise from the cortex of the root in Podostemaceae. Therefore, comparative analysis of the root-borne shoot formation between Hypericaceae and Podostemaceae is needed to understand the emergence of the unique body plan in Podostemaceae.
P0646 – ePoster
Structural mechanisms of the development and
1 germination of seeds with underdeveloped embryo
Kravkina, I 1 , Koteyeva, N 1
Komarov Botanical Institute RAS, Russia
Embryo underdevelopment in mature seed is very often linked with physiological dormancy both at dicotyledonous and monocotyledonous plants. Such seeds need specific temperature conditions during embryo development and seed germination. The purpose of the work was to study structural reactions of seeds to variable temperature conditions during embryo development and germination in the Tulipa tarda .
Comparative studies of the structural changes were carried out at the embryo cotyledon and root tip cells, as well as at the endosperm cells using light and transmission electron microscopy. The process of embryo development and initial stages of the seed germination were studied under warm stratification at the temperature +22±1ºC, under cold stratification at the temperature +3 ± 1ºC, and under their sequential changes. It was shown that as early as after 24h of seed imbibitions in all investigated tissues and under all conditions the fast structural reactions took place confirming the metabolism activation. Probably, in the seed tissues metabolism activation occurs under certain wateriness regardless of dormancy type and stratification conditions. Increase of cotyledon cells sizes during embryo development in all conditions occurs only because of active syntheses of the cells structural components. The critical moment of the embryo development is a reaching of 3.2 mm in length when under the warm stratification embryo stops to growth
(20–25 days), while under cold stratification (40–45 days) divisions of the cells begin at cotyledon base. First signs of mobilization of reserved substance in endosperm

are observed at the same stage. This indicates clear relationship between the beginning of reserve mobilization in endosperm and proliferation process in embryo cotyledons, and let us to suppose more significant role of embryo, than endosperm during embryo development. Primary structure formation of the radicle begins with differentiation of the cap cells. The first signs of differentiation are observed only under cold stratification on proliferation stage of the cotyledon base cells. To complete of embryo development (only under cold temperature), embryo root is clearly differentiated on the group of meristematic cells and root cap. The embryo growth is not accompanied by increase of the embryo root sizes until germination. Growth of seedling root after germination occurs only after increase of the temperature. These indicate that cold stratification which release morpho-physiologycal dormancy and initiates germination, inhibits root meristem proliferation after germination occurs. In summary, the germination of T. tarda seeds is a temperature dependent process, which triggers by a fast extension of tabular cells at the cotyledon base. Analysis of data obtained shows that processes of extension and proliferation of embryo cells during development and germination are independent and initiated by different temperature conditions.
P0648 – ePoster
Structure, development and histochemistry of the bracteal and receptacular colleters of
Encyclia alboxanthina
(Orchidaceae)
Leitão, CA 1 , Ascensão, L 2
1 Universidade Estadual do Sudoeste da Bahia, DCN,
2 Vitória da Conquista, Brazil; Universidade de Lisboa,
Faculdade de Ciências, Departamento de Biologia
Vegetal, Lisboa, Portugal
Orchidaceae family is notorious by the complex and diverse morphology of vegetative and reproductive organs. Although many studies have been conducted on the morphology of orchids, the occurrence of colleters has been only recently reported in Rodriguezia venusta, an epiphytic orchid native to Brazil and Peru. Colleters are bud characteristic external secretory structures, that produce a sticky secretion to protect the developing meristems and young foliar and floral organs. In the last years, these structures have been described in many species of a large number of dicot families and, considering its occurrence in R. venusta, it seems reasonable to assume that colleters are also present in other Orchidaceae. In this study we describe the morphology, development and histochemistry of the colleters of Encyclia alboxanthina , a terrestrial orchid, apparently endemic to Chapada Diamantina (Bahia State,
Brazil). Flower buds, at different stages of development, as well as, the subtending bracts were collected from natural populations and fixed with Karnovsky. The samples were prepared for SEM or embedding in Leica
Historesin following the standard methods. Sections of the embedded tissues were cut and stained for general histology and to localize in situ the main chemical classes of compounds present in the secretion. The colleters of E. alboxanthina occur on the adaxial surface of the inflorescence bracts and on floral receptacle. Both types are multicellular, uniseriate trichomes with a finger-like form. However, they differentiate at different time on the course of bud development, they have a different number of cells and the secretion is also quantitatively and qualitatively different. The bracteal colleters begins to develop in very young floral buds, with less than 1mm long (measure excluding the pedicel) and are constituted by a basal/stalk cell and one to three head cells in line. The receptacular colleters develop later in buds with 5mm long and have a basal/stalk cell and four head cells. Both originate from a hypertrophied protodermal cell that undergoes sequential periclinal divisions and their secretory activity begins even at the stage of cell expansion. Apparently, bracteal colleters stopped early their secretory activity. In floral buds 3mm long, they seem collapsed, showing cytoplasm retraction in the head cells and, in recently-opened flowers, 20mm long, they are at an advanced stage of senescence. By contrast, receptacular colleters seem active until the flower anthesis. Bracteal colleters produce an abundant secretion with a heterogeneous aspect. The secretion products stained intensively with PAS and tannic acid/ferric chloride for polysaccharides and mucilage, respectively, but did not react with Alcian blue for acidic polysaccharides. Toluidine blue O, Xylidine ponceau and
PAS also stained positively the trichome head cells.
Unlike the bracteal colleters, the receptacular produce only a small amount of secretion that was detected by
PAS over the trichome surfaces, in floral buds 15mm long. Bracteal colleters probably facilitate the young and fragile bud sliding over the bract surface and the mucilaginous secretion could act as a lubricant in this process. However, future research is necessary to understanding the role of the receptacular colleters on this orchid. CAPES.
P0649 – ePoster
1
Geotropic orientation of a lateral bud of monocotyledons
Levichev, I 1
Komarov Botanical Institute, Russia
The positive geotropism of a root is well-known. The lateral bud appeared to be a geotropically oriented structure originally and is homologous to a root in terms of position of its initial. This conclusion is empirically confirmed and arises from the system analysis of a probable algorithm of occurrence of cormophyta and angiosperma as a result of evolution of a bipolar telome with no internal contradictions. The elementary phytomerical module of monocotyledons is a complex bipolar structure, a dimeric (mono-sympodial) homolog of a mesom, possesses an axial field (internode) with four growth initials (two at the top and two at the bottom) according to the known and newly ascertained data collection. An overland emergence of plants required a mandatory transition to a dichopodial principle of growing and branching. The internode of dichopodial nature of the elementary phytomerical module predominately develops an appendicular obligatively final (never continuing) leaf (arisen from a single lateral sporangium) out of the top ageotropic initial. The second
(inhibited) ageotropic initial gives a rise monopodially
(dichopodially would be more correct) to the subsequent phytomerical module. The antipodes of upper initials are
543

the polar geotropic initials at the lower end of the internode. The initial under a leaf (as well as under a cotyledon) predominately develops a root (exogenously at the starting module, i.e. an embryo). The vascular connection and an emergence by the ataktostela elements to the next additional root of the endogenous nature are in each shoot metamere located above. A lateral bud is formed from the opposite side relative to the root (and relative to the leaf at the top end of the internode) out of the second inhibited geotropic initial. Thus, the fundamental difference between monopodial and sympodial continuation of any shoot of monocotyledons consists not only in the top and bottom position of growth initials, respectively, but also in their initial polar multidirectional growth. As these initials are the optical antipodes, their prophylls are faced outside with the dorsal sides, and their ventral surfaces are directed towards continuation of the axial body, i.e. the hypopodium. The sympodial photosynthesizing shoot is forced to change the ancestrally geotropic orientation of growth to the ageotropic orientation. As a result, a unique orientation of the sympodium prophyll occurs: the addorsed (à dos, French) position with the dorsal surface
(abaxial surface) faced to the parent axis, which is often accompanied by an occurrence of two keels in a prophyll.
Two keels represent an obvious, but not a compulsory attribute which depends on many circumstances of the morphogenesis, while dimerism is the cause of this phenomenon. The starting phytomer, i.e. the seedling never has a lateral bud, as the area of a root collar is not protected. The lateral buds above are securely covered with a leaf from the lower metamere and are located in its axil, although they belong to and are contiguously formed by an overlying metamere. The nature of a lateral bud of monocotyledons fundamentally differs from the nature of an axillary bud of dicotyledons.
P0650 – ePoster
Dimeric, mono-sympodial, structure of metameric module of agiosperms
Levichev, I 1 , Krasovskaya, L 1
1 Komarov Botanical Institute, Russia
An alternative hypothesis for angiosperm shoot structure arrangement is offered to supplement statements of the telome theory with the evolutionary series: telome, mesom, phytomer, phytodimer, phytopolymer, and to postulate an occurrence of angiosperma as a consequence of an appearance of the dimeric type metamere (with its locking as a starting metamere) following the dorsiventral congenital integration of parent monopodial and daughter sympodial structures. The mono-sympodial integration is supported by the following: double tunicle, bidirectional meristematic activity of a leaf primordium, median twin-cone of certain leaf types and sympodial prophyll, epiblast and coleorhiza, conjugated and central stipules, ligula, antiligula, hastula, axil on a chard, rudimentary appendage of an arum family prophyll, slant internodes of bamboos, white peduncle coloration in the taxons with white central region of the leaf, etc. This occurrence (angiosperma appearance) takes a defined place in the evolutionary range of cormophyta formation preceding a separation into monocotyledons and dicotyledons; supplements and enriches interpretation of
544 phenomena currently devoid of logically and evidentially substantiated explanations: one and two cotyledons, scutellum of cereals, true leaf (macrophyll), lateral position on the shoot apex of leaf primordia, division into an axil, chard, true leaf blade, growing and branching, monopodium and sympodium, addorsed orientation of the sympodium prophyll and presence of two keels in it, phytomer and phyllorhiza, storied display of neoteny on a shoot, polar opposite orientation of a shoot and rootage, metamerism, modularity, coloniality, distinction of starting metameres, i.e. angiosperm embryos and their arrangement into a superpositional phytomer strand in monocotyledons and a superpositional strand of sheafs
(of phyllorhizas in fact, i.e. a juxtaposition integration of phytomers) in dicotyledons, etc. An angiosperma specific double tunicle ovule as a consequence of module dimerism (congenital conjugation in a dorsiventral sequence of monopodium and sympodium) became possible as a result of implementation of a bipolar telome start (1) and the following mesom (2) evolution during land development performed by plant bodies. This was promoted by an appearance of a root as a consequence of symbiogenesis with fungi (3), occurrence of an adventitious and permanently final leaf blade out of sporangium (4), transformation of dichotomy into dichopodiality (5) compulsory for land plants, advantages of polyvariance of degrees of freedom of growth processes with bipolar growing and branching according to the dichopodial principle (6), improvement of mesom- and then phytomer-arranged elementary cormophytic structures (7) of module-colonial soma into phytodi- and phytopolymer (8) structures of angiosperma and iterative participation of storied neoteny (9) in these evolutionary transformations.
P0651 – ePoster
1
Singularity and differences between monocotyledons and dicotyledons
Levichev, I 1 , Krasovskaya, L 1 , Tatanov, I
Komarov Botanical Institute, Russia
1
In addition to the distinction in cotyledons number, an absence of the lateral bud external display is typical for dicotyledons, while an axillary bud of radically different nature is surely present in them. The following is in common for these two classes: bipolar colonially phytomeric structure of soma with a linearly and ageotropically oriented shoot which grows metamerically, sequentially and superpositionally with dimeric modules (mono-sympodial homologs of a mesom) of various dimension in the form of a strand.
Owing to the symbiosogenic nature of such soma, the fungi component participates in geotropic formation of conducting system and rootage. The following evolutionary algorithm of occurrence of cormophyta and angiosperma is probable: the single telome axis is bipolarly transformed into a 5-axis mesom. The dichopodial type neotenical aromorphosis of a mesom with a single lateral sporangium was formed into a cormophytical phytomer. The latter, having combined a parent monopodium and a daughter sympodium in the dorsiventral direction, has formed an elementary dimer with a conjoint geotropic body, i.e. a root. Integration of two dimers (for example, in an embryo sac with the

polyembryony) resulted in occurrence of a dicotyledon phyllorhiza. This is an indicative fact that the dimeric integration (alias the mono-sympodial integration) in no way breaks a phyllotaxis of the phytomerical module neither in monocotyledons, nor in dicotyledons.
Combination of mono- and sympodial structures in a single starting metamere, i.e. an embryo, occurs out of different mesom number – 2 in monocotyledons and 4 in dicotyledons, and accepts different quantity of telome axes: in monocotyledons – 9 (evidentially), while 7 empirically in the vegetative module and 6 in the starting phytodimer (an embryo; the lateral bud is absent); in dicotyledons – 18 (evidentially), while 13 empirically in both vegetative and starting phyllorhizic modules. The modules in the ontogenesis are replicated by the structural similarity into a taxon colonial individual (an integral organism) of the specific class (the reproductive type modules are not considered now). The phytodimeric modules with another quantity of accepted axes which are not considered now as well occur in monocotyledons
(especially in the arum family) and in dicotyledons
(especially in the tropical plants). The shoot arrangement differences in these two classes are reduced to the following: in monocotyledons – the phytomerical modules ageotropically grow both monopodially
(dichopodially would be more correct) from the upper initial and sympodially (which provides branching) from the geotropically oriented (the prophyll is therefore addorsed) lateral initial. The exogenous annual primary root and endogenous additional roots are formed with the geotropic growth coming out always above the primary root collar; in dicotyledons – the ageotropic growth occurs in the form of a strand of sheaves out of juxtapositionally integrated phyllorhizic modules sympodially – from 1 or from 2 (which is rarer) lower initials, or in the form of branches out of axillary buds. In case of the geotropic growth, an exogenous, usually perennial primary branching root.
P0652 – ePoster
An anther-specific gene encoding Cisprenyltransferase in Lilium longiflorum
, Wang, C-S 1
1
Liu, M-C 1
Graduate Institute of Biotechnology, NCHU, Taichung
A cis-prenyltransferase gene, LLA66, was identified from a suppression subtractive cDNA library at the phase of microspore development in the anther of Lilium longiflorum . The method of 5’-RACE-PCR was used to obtain the full-length of LLA66 cDNA (accession no.
DQ911525) with a size of 1151 bp, excluding the poly
(A) tail. The LLA66 cDNA contains an open reading frame of 927 bp encoding a polypeptide of 308 amino acids with a calculated molecular mass of 35.7 kDa.
Sequence alignment revealed that the LLA66 protein shares 30-41% identity (42-56% similarity) with cisprenyltransferases of various species and is phylogenetically distinct from other monocot cisprenyltransferases. The protein is highly conserved in five regions of cis-prenyltransferases that are thought to be important for the substrate binding and catalytic function. RNA blot analysis indicated that the LLA66 gene was anther specific and specifically expressed at the phase of microspore development in the anther. In situ hybridization with DIG-labeled antisense riboprobe of the LLA66 showed that strong signals were detected at the tapetal layer of the anther wall, indicating that the
LLA66 mRNA was synthesized in tapetum. The LLA66 mRNA was also detected in the microspore. RNA blot indicated that the LLA66 gene was endogenously induced by gibberellin (GA). However it cannot be induced with exogenous addition of GA. The enzyme activity of prenyltransferases in various stages of anther buds correlates both with the tapetal growth and disintegration and microspore development. The coding region of LLA66 constructed in pYES2 vector was introduced into S. cerevisiae (INVSc1). The LLA66 protein was expressed and affinity-purified using Ni2 + nitrilotriacetic acid agarose. In vitro enzymatic assay revealed that the enzyme catalyzed the formation of polyprenyl diphosphates. A drastic increase of enzyme activity was detected with the addition of Mg2 +
1 mM. A further addition of Mg2 +
ion up to
ion inhibited the enzyme activity. The optimum pH value is determined to be pH 8.0. We propose that cis-prenyltransferase in the tapetum and microspore may involve in the synthesis of dolichols and polyprenols compounds that may facilitate the wall formation of microspores.
P0653 – ePoster
Organelle tracking during environmentally induced and developmentally regulated programmed cell death (PCD) in the model species
Aponogeton
1 madagascariensis
Lord, C 1 , Gunawardena, A 1
Dalhousie University, Canada
The lace plant, Aponogeton madagascariensis is an aquatic monocot, which forms perforations in its leaves between longitudinal and transverse veins, through developmentally regulated programmed cell death
(PCD). Developmental PCD is a genetically encoded process, which is involved throughout normal development in plant life cycles. Within plant systems a second form of PCD exists known as environmentally regulated PCD; conversely to developmentally regulated
PCD, environmentally induced PCD is not a genetically encoded process and occurs as a result of external biotic or abiotic factors. Many questions regarding the differences and similarities between developmentally regulated and environmentally induced cell death are still unanswered, and a time line for organelle changes in both forms of cell death in plant systems is still unclear. To better elucidate differences and similarities between the two forms of PCD, protoplasts isolated from control areas of the lace plant were heat shocked at 55°C for 20 minutes to induce PCD. Following heat induced PCD organelles including the mitochondria, chloroplasts, nucleus, vesicles and cytoskeleton were examined via long term live cell imaging and confocal microscopy to better understand organelle characteristics including origin, movement and fate. This organelle tracking is also currently under investigation in whole lace plant leaves focusing on the regulated form of PCD. Organelle characteristics will then be compared between the developmentally regulated and environmentally induced forms of cell death. This examination of organelles will allow for the first ever opportunity to compare both
545

forms of PCD within one species of plant. Interestingly, initial results show numerous characteristics of developmental PCD in the heat-induced system, including plasma membrane blebbing, vesicle formation, dramatic mitochondrial and cytoskeleton changes, and fragmented DNA detected via TUNEL. Thus far, many organelle changes in the induced system appear to be shared with the developmentally regulated system.
Further experiments are required in order to characterize each organelle on an exceedingly in depth level and to create a detailed time line of cellular changes for both forms of cell death in one species.
P0654 – ePoster
Diversity of pollen, pollination and conservation
Mandal, S 1
1 Visva-Bharati University, India
Pollen, the male partner in the fertilization process of flowering plants, provides an excellent material for several physiological and biochemical studies of fundamental and applied nature. Pollination is a basic force for gene recombination in flowering plants plays a key role in plant breeding programmes. Pollination, pollen germination and stigma receptivity must be analysed critically on a species by species basis, as it reflects the basic criteria for breeding programmes. As such, a detailed knowledge about pollination, pollen germination and stigma receptivity will be helpful to produce genetically superior stocks. The present paper will highlight the recent problems and future prospects of plant and pollinator resources of some tropical plants growing in India in the context of environmental degradation and global warming for biodiversity conservation. Globally, of the estimated 1330 crop plants grown for food, beverages, fibers, spices and medicines, approximately 1000 (75%) are pollinated by animals. It has been calculated that pollinators deliver one out of every three mouthfuls of food we eat, and beverages we drink. The study of pollinator decline is also interesting to the scientists, as pollinators have the potential to become a keystone indicator of environmental degradation. The plant-pollinator interactions have contributed to a paradigm shift from protecting individual species to protecting inter-specific relationships and landscape-level ecological processes.
The ultimate goal of the pollinator conservation system is to ensure healthy and self-sustaining populations of pollinator resources.
P0655 – ePoster
ABA-induced reorientation of cortical microtubule
1 arrays involves the formation of dynamic, discordant microtubules
Marc, J 1 , Seung, D 1 , Webster, M
University of Sydney, Australia
1
Abscisic acid (ABA) is involved in plant development and responses to environmental stress including the formation of longitudinal microtubule arrays in elongating cells, although the underlying mechanism is
546 unknown. We explored ABA-induced microtubule reorientation in leek (Allium porrum L.) leaf epidermal cells transiently expressing a GFP¡VMBD microtubule reporter. After 14–18 h incubation with ABA, the frequency of cells with longitudinal arrays of cortical microtubules along the outer epidermal wall increased with dose-dependency until saturation at 20
µ
M. Timelapse imaging of individual cells revealed a gradual increase in the occurrence of discordant, dynamic microtubules deviating from the normal transverse microtubule array within 2–4 h of exposure to ABA, followed by reorientation into a completely longitudinal array within 8 h. Reorientation occurred also in epidermal cells in Arabidopsis leaves and in the elongation zone of root tips. The formation of discordant microtubules in Allium was unaffected by inhibiting cytoplasmic streaming with Latrunculin B. However, transient expression of AtEB1b-GFP reporter and analysis of 'comet' velocities revealed that the microtubule growth rate increased within 2 h of exposure to ABA, with an average increase of ~21% over 8 h relative to controls. ABA also increased the sensitivity of microtubules to depolymerization by oryzalin and exacerbated oryzalin-induced radial swelling of
Arabidopsis root tips. The swelling was further aggravated in AtPLD
δ
-null mutant, suggesting PLD
δ plays a role in microtubule stability. We propose that
ABA-induced reorientation of transverse microtubule array initially involves destabilization of the array combined with the formation of dynamic, discordant microtubules.
P0656 – ePoster
Microsporogenesis and microgametogenesis of
Bromeliaceae from southern Brazil: pollen fertility in conditions of ex situ conservation
Mariath, J 1,3 , Duarte-Silva, E 4 de Andrade, G 2
, Troleis, J 1 , Pesenti, A 1 ,
1 Universidade Federal do Rio Grande do Sul, Porto
Alegre, RS, Brazil; 2 Universidade Federal de Viçosa,
Minas Gerais, Brazil; 3 CNPq researcher; 4 CAPES fellowship
Ex-situ conservation means ‘off-site’ conservation and it is a process to protect endangered species outside its natural habitat. Several Bromeliaceae species are endangered of extinction, including Dickya distachya from Southern Brazil. Individuals of Dickya brevifolia ,
Dickya distachya and Vriesea gigantea collected in natural habitats of Rio Grande do Sul state, Brazil are kept in scientific collections in Porto Alegre – RS, Brazil.
Their microsporogenesis and microgametogenesis were investigated to verify the pollen fertility in conditions of ex-situ conservation. Microscopical analyses were made with anthers in different stages of development, fixed in
1% glutaraldehyde and 4% formaldehyde, treated with
0.1M sodium phosphate buffer pH7.2, dehydrated in ethylic series and included in hydroxyethylmetacrylate.
Slices of 2-5µm were stained with 0.05% Toluidine Blue
O pH4.4. Observations and photomicrographs were made using Leica DMR-HC with a DFC-500 digital camera.
As results, microsporogenesis and microgametogenesis of D. brevifolia and V. gigantea were regular in all stages observed while D. distachya analyses shown

abnormalities at microgametogenesis. In D. brevifolia and V. gigantea , microspore mother cells are isolated by callosis and start meiosis-I. The cytokinesis is sucessive.
Microspore dyads start meiosis II and give rise to tetrahedrical tetrads, and, in some cases, isobilateral.
Observed tetrads are regular and present one nucleous per cell. Micronucleous or microcytes are not observed.
After callosis dissolution, sporopollenin is deposited in microspore primexine matrix and microspores become free at locule. Microspore vacuolate and, after mitosis, gives rise to a bicellular polen. Vegetative cell nucleous embraces generative cell. Pollen grain is dispersed in bicelular form. At this stage, all pollen observed were in monads and malformated tetrads do not occur. In V. gigantea , some pollen tubes germinate in the anther.
Analysis of D. distachya shown anormalities at the end of microgametogenesis. At bicelular pollen stage, after the complete formation of sporodermis, with intine and exine, abnormal mitosis occurs and gives rise to three or four-celled pollen grains. In both cases, vegetative cell undergoes a symetric mitosis. Three-celled pollen bears the generative cell and two symetric big cells with conspicuos nucleous, originated by the vegetative cell. In four-celled pollen, both vegetative and generative cell undergo mitosis, forming two big cells with conspicuos nucleus, originated by the vegetative cell and two sperm cells at the periphery of pollen grain. We conclude that
D. brevifolia and V. gigantea showed regular pollen grains in ex-situ conservation condition. About pollen fertility of D. distachya , more studies are necessary. As perspectives, tests of pollen viability, including pollen germination will be made. Financial support: CAPES and
CNPq.
P0657 – ePoster
The
Passiflora
ovule: from the external to internal view
Mariath, J 1,3 , Silverio, A 4 , de Souza, G 2
1 Universidade Federal do Rio Grande do Sul, Porto
Alegre, RS, Brazil; 2 Programa de Pós-Graduação em
Botânica, Biociências, UFRGS, Brazil; researcher; 4 CAPES fellowship
3 CNPq
This present study was carried out with Passiflora species from South America. Passifloraceae has plants with diverse habits: trees, shrubs, herbs, and particularly to Passiflora L. genus, a climbers plant with a high vegetative and floral morphological diversity, are still mostly unknown in their reproductive anatomical features, mainly at the embryological level. Information about the structural morphology in reproductive organs, such as mature ovules, are based on classical studies and has been made in few species with economical use, neglecting possible variations in the anatomical architecture of wild plants. This study aimed to describe the internal and external morphological structure of mature ovules from twelve species of Passiflora – P. alata Curtis, P. amethystina Mikan, P. caerulea L., P. edulis Sims, P. elegans Mast. and P. tenuifila Killip
( Passiflora Clade), P. capsularis L., P. misera Kunth, P. morifolia Mast. and P. suberosa L. ( Decaloba Clade), P. haematostigma Mart. ex Mast. ( Astrophea Clade) and P. ovalis Vell. ex M. Roem. ( Deidamioides Clade), contributing to the knowledge of the genus reproductive biology. Ovules were included in hydroxyethylmethacrylate, sliced in 4–6µm, stained with
Toluidine Blue O, and examined in bright-field with an
Leica DMR-HC. For scanning electron microscopy the samples were critical point dried and coated with gold in a sputtering system, and analysed in a JEOL 6060. The ovules are anatropous, arillated, bitegmic and their micropiles are arranged in zig-zag. The ovules are crassinucellate and the integuments are originated in the dermal layer. The nucellar epidermis divides periclinally on the apical portion and show differences in this nucellar cap, between the analyzed species, and are partially degraded at the mature stage. The gametogenesis has the Polygonum type of development and the mature gametophyte has ephemeral antipodal cells and two synergids with a developed filiform apparatus that can show different features among the species. Financial support: CAPES and CNPq.
P0658 – ePoster
Development of root aerenchyma in
Potamogeton polygonus
Cham. et Schltdl (Potamogetonaceae –
Alismatales): cell wall structure and cytological alterations
Mastroberti, A 1 , Moço, MC 1 , Bona, C 2 , Mariath, JE 1
1 Federal University of Rio Grande do Sul, Brazil;
2 Federal University of Parana, Brazil
The formation of lysigenous aerenchyma through the collapse of cells and maintaining cell wall integrity is a common process among aquatic plants. Literature describes four types of lysigenous aerenchyma: lysigenous 'in packages,' schizo-lysigenous, radial and tangential lysigenous. The survival of some cells in the cortical aerenchyma has an important role in the integrity of root structure and nutrient transport via symplastic or apoplastic pathways. Studies on lysigenous aerenchyma formation are based on model plants such as Zea mays
(maize). In this species there are changes in the level of methyl-esterification of pectins in the cells forming the aerenchyma when subjected to hypoxia and ethylene. It is known that changes in specific epitopes in the pectic molecule and hemicelluloses are important to the functional properties of the wall, including structural properties and cell adhesion. Such features are essential for studies on aerenchyma ontogenesis. Potamogeton polygonus is an aquatic plant. The roots present aerenchyma classified as radial lysigenous, formed by radial rows of collapsed cortical cells. Cytological data, including the cell wall dynamics of collapsed cells in the aerenchyma, may provide answers about root organogenesis. The aim of this work was to analyse the cytological cell changes that form air spaces during root ontogeny. Cortical cells were studied at two developmental stages: stage 1, cross-section at the region of elongation, and stage 2, cross-section at the absorption zone. The material was processed for light microscopy, immunofluorescence using monoclonal antibodies to recognize different pectic epitopes (LM19, LM20, LM5,
LM6) and hemicelluloses such as xyloglucans (LM15) and (galacto)(gluco)mannans (LM21), and transmission electron microscopy. Those cells that form aerenchyma undergo a gradual degradation of organelles. The middle lamella is persistent and the primary cell wall is thicker
547

in the cell adhesion regions. Pectins with low methylesterification (LM19) occurred throughout the cell wall and middle lamella. Arabinans (LM6) were not observed.
However, methyl-esterified pectin (LM20) and galactans
(LM5) were present only in the areas of cell adhesion.
De-esterified pectin contributes to a more resistant gel and is associated to cell separation in the formation of intercellular spaces, maintaining the middle lamella and cell wall integrity. Xyloglucans were less evident in the cells forming the aerenchyma. Surprisingly, mannans are the most abundant hemicellulose in the analysed stages.
Cell wall mannans operate as storage and structural polysaccharides. Compared with xyloglucans, little is known about the role of mannans in the cell wall. It has been described that mannans appear to be less abundant in the primary cell wall of angiosperms. P. polygonus is a basal monocotyledon and the chemical features of the cell wall are different when compared with dicotyledons or commelinoid monocotyledons. The structural role and its interaction with cellulose resemble that of xyloglucan.
Mannans may determine the firmness and flexibility of this tissue. In conclusion, schizogeny is involved in aerenchyma formation because the changes in the esterification of pectins is not followed by cell lysis, and the wide presence of mannans as structural hemicellulose, instead of xyloglucans, could maintain cell integrity.
P0660 – ePoster
To break or not to break? It's what's inside that counts!
Meicenheimer, RD 1 , Coffin, DW 2 , Chapman, EM 1
1 Dept of Botany, Miami University, USA; 2 Dept of Paper and Chemical Engineering, Miami University, USA
Differences in the flexibility of Pinus nigra and Pinus resinosa leaves can be used to discriminate these two similarly looking pine species from one another. When bent along the longitudinal axis, P. resinosa leaves 'snap', while P. nigra leaves appear 'flexible'. This useful field test has had no known biophysical or anatomical explanation until now. Flexible leaves of P. nigra fail in compression, while brittle P. resinosa leaves fail in tension when subject to longitudinal bending. First order, mechanical analysis of bending and buckling was applied to the pine needles to elucidate the important anatomical differences between these two species, which can account for their different biophysical behavior when subject to bending. There was no significant difference in the cross section of the total leaf area, or the inner core
(endodermis + transfusion tissue + vascular bundles) area between the two species. P. nigra had a thicker outer core (epidermis + hypodermis), but this could not account for the differences in the biophysical behavior of the two species leaves. Our investigation revealed that it was differences in the pattern of cell wall thickening and lignification of the endodermal layer of the inner core of the leaves that best explains the differences between bending behavior of the two species.
P0661 – ePoster
New reports of colleters in Myrtaceae
548
1
Meira, CJ 1 , Barbosa, LCA 2 , Meira, RMSA 1
Departamento de Biologia Vegetal, Universidade
2
Federal de Viçosa, Minas Gerais, Brazil;
Departamento de Química, Universidade Federal de
Viçosa, Brazil
Colleters are multicellular secretory structures that may occur in vegetative or reproductive shoot apices of many botany families. However, in the Myrtales Order, reports of colleters are fragmented and addressed only in the morphological aspect. This is the first report of occurrence of colleters in Myrtaceae family. Colleters from species of the Leptospermoide and Myrtoideae subfamilies were studied regarding its morphology and anatomy. Shoot meristems were collected, fixed in formalin-acetic-50% ethanol (1:1:18 v/v), and embedded in methacrylate or observed on scanning electronic microscopy. Serial cross and longitudinal sections were on an automatic rotary microtome, stained with Toluidine blue, and mounted in synthetic resin. Some fresh samples were subjected to Periodic-Acid-Schiff’s reaction for detection of neutral polysaccharides, and Ruthenium Red or tanic acid-ferric chloride for pectins and mucilaginous substances. We have observed colleters in 15 species from seven genera ( Callistemon, Campomanesia,
Eugenia, Leptospermum, Melaleuca, Myrciaria,
Psidium ). The colleters are morphologically and anatomically different from the types described for other families. As a result, we have classified and described three new colleters types: the petaloid type, in cross section, presents an axis that is flattened dorsiventrally, much wider than thick, is short, and usually occurs associated with cataphylls that involve the apical buds of vegetative shoots in the dry season; the conic type has an axis that is seen in cross section as a circular or ellipsoid form with decreasing diameter towards the apex, can be sessile or with short stalk; the euriform type that seems the petaloid type, is flattened dorsiventrally in cross section and its axis is much wider than thick, longer than wide. These three colleters type didn´t occur all together in all species. The petaloid colleters were present only in the Callistemon and Melaleuca species from
Leptospermoideae subfamily. Both genera are phylogenetically close related and these results contribute with the taxonomic argue that they should be only one genera. On the other hand, the euriform and conic colleters occurs in both, Leptospermoideae and
Myrtoideae subfamilies, an evidence of phylogenetical relation. The mucilaginous secretion has been confirmed by histochemical tests. The secretion analysis by electrophoresis method (SDS-PAGE) has revealed no proteins in the Myrtaceae colleters’ secretion. Therefore, there is no evidence of protection against microorganisms. Thus, the main function of the colleters’ secretion from Myrtaceae species are to protect plants from potentially damaging abiotic agents as low relative humidity and high levels of solar radiation which are capable of dehydrating young leaves and other undifferentiated plant tissue. The Myrtaceae's colleters exhibit a potential taxonomic application once they can contribute to clarify the phylogenetic relationships of
Myrtaceae family. (FAPEMIG, CNPq).

P0664 – ePoster
The diversity of pollen type and sculptural structure in family Leguminosae : implication for evolution
Nakkuntod, M 1 , Seangruttanachaikul, I 1 , Jabthong, R 1
1 Dept of Biology, Faculty of Science, Naresuan
University, Phitsanulok, Thailand
The pollen morphology of 32 species representing 22 genera of the family Leguminosae, divided into 3 subfamilies: Mimosoideae, Caesalpinioideae and
Papilionoideae. Pollen morphology was examined by light and scanning electron microscopes. The pollen grains are generally radial symmetry, isopolar, mostly tricolporate except Vigna unguiculata is tripolate operculate. The exine sculpturing is commonly reticulate or subpsilate and sometimes striate. There are 3 pollen types in subfamily Mimosoideae: monad, tetrads and polyads. Subfamily Caesalpinioideae is classified into 3 groups depending on sculptural structure: striate, reticulate and subpsilate. Subfamily Papilionoideae is divided into 2 groups up to pollen shape: suboblate and prolate spheroidal. The evolution of these subfamilies showed that subfamily Mimosoideae is the most primitive. The pollen morphology of this family is significantly useful at generic and specific levels for classification and reveals their evolution.
P0665 – ePoster
Nectar and nectary character state
Nepi, M 1 , Guarnieri, M 1 , Nocentini, D 1 , Pacini, E 1
1 University of Siena, Dept of Environmental Sciences,
Italy
Nectar is a fluid released by a secretory tissue, the nectary, to favour benevolent relationships with animals.
The type of benevolence depends on nectary location.
Nectaries located in the flower are termed floral and the nectar attracts pollinators. Nectaries located on leaves or green stems are termed extrafloral and the nectar rewards ants that discourage predators. Nectary anatomy is organized as follows: a. epidermis surmounted by cuticle, with or without stomata; b. parenchyma having cells bearing chloroplasts or amyloplasts; c. sub parenchyma (optional); d. vascular bundles composed of phloem and xylem but in some cases only phloem.
Nectary parenchyma with chloroplasts is the more common type and always the case in extrafloral nectaries. These cytological features determine when and how the carbohydrate fraction of nectar is produced, when nectar is secreted and at what rate and concentration. Variations in nectar components determine differences in nectary structure, how nectar is secreted and some aspects of nectar composition. The secreted nectar may maintain a constant volume, especially if nectary parenchyma has amyloplasts, or may vary according to how much is collected by consumers; feedback has been documented. The nectary may be a single structure or consist of separate secretory trichomes. Nectar may be released over short periods (a few hours) or for longer periods (days or even months in the case of extrafloral nectaries). Floral nectaries may be situated at the ovary surface, or recessed into the ovary or other flower parts; in the latter case, nectar flows out through microchannels . The nectary epidermis or all its parts may degenerate after secreting or persist and continue photosynthesis to nourish seed/fruit growth.
Reabsorption of uncollected nectar at the end of flower receptivity occurs in many species, especially if much nectar is secreted. The term nectar presentation indicates how nectar is offered to consumers. Primary presentation is when nectar is deposited on the surface of the nectary, or very close to it. Secondary presentation is when nectar is secreted by a nectary located in the central part of the flower, i.e. the receptacle, but flows out and is deposited elsewhere for consumption, as in spurs. Nectar is an aqueous solution of ions, carbohydrates, amino acids, enzymes, antibiotics, antioxidants and occasionally lipids. These substances may be rewards for consumers and may maintain nectar homeostasis (composition and viscosity), preventing the growth of airborne or vectorborne spores and fungi. Constant viscosity ensures that the nectar remains available to animals having a given type of mouth-part.
P0666 – ePoster
Microscopic characterization of foliar secretory trichomes of
Drosera capensis
Naidoo, Y 1 , Nicholas, A 1 , Ibrahim, S 2
1 University KwaZulu Natal - Westville campus, South
Africa; 2 Suez Canal University, Egypt
The carnivorous habit has been described in over 600 different plant species from a total of 300,000 species of vascular plants. These plants are typically found inhabiting soils that are poor in nutrients, especially nitrogen hence carnivory is one of many possible adaptation strategies. Drosera L., commonly known as the ‘Sundews’, comprises the second largest genera of carnivorous plants, with over 170 species. Drosera capensis is fairly common in the south Western Cape,
South Africa, hence the name ‘Cape sundew’. Plants of this genus use an active method of attraction and entrapment. The trapping and digestion mechanism of D. capensis employ remarkable glandular activity whereby stalked and sessile glands secrete mucilage to attract and ensnare insects and enzymes to digest them. The present study concerns a micromorphological investigation of the foliar glandular trichomes of D. capensis . Segments of leaf material of D. capensis were prepared for scanning and transmission electron microscopy (SEM & TEM) using conventional methods. D. capensis produces congested, strap-like leaves that are elevated from the ground surface. Stalked filamentous trichomes form a dense fringe along the margin of the leaf while fewer, shorter filamentous trichomes occur towards the centre of the blade. There are predominantly 3 types of glandular trichomes on both adaxial and abaxial surfaces. These include sessile-capitate and long-stalked capitate trichomes. The long stalked glands are biseriate and sparsely distributed relative to the filamentous and sessile trichomes. TEM of the sessile-capitate trichomes showed the presence of wall ingrowths in the form of irregular dendritic projections initiated at the junction of the outer tangential and radial cell walls and developing centripetally along the inner tangential wall. These
549

ingrowths conform to the shape of the invaginated plasmalemma. Other cytoplasmic features include abundant ribosomes, electron-dense vesicles that occur in close proximity to the nucleus and plasmalemma especially in the basal cells of the gland. The trapping and digestion mechanism of D. capensis employ remarkable glandular activity whereby stalked and sessile glands secrete mucilage to attract and ensnare insects and enzymes to digest them. It was reported that Type I and
IV glands feature in D. capensis . Type I is the most simple and common type called a papilla equivalent to the capitate-sessile trichomes in D.capensis. Type IV glands are unbranched glandular hairs with a biseriate stalk equivalent to the long-stalked capitate trichomes.
These glands are presumed to be exclusively involved in the secretion of digestive enzymes and in the absorption of insect-derived nutrients. Wall ingrowths a characteristic of transfer cells occur in the basal cells of sessile trichomes in D. capensis . However, wall ingrowths have been reported in the filaments of Drosera, stalked and sessile glands of Pinguicula, Drosophyllum,
Dionea and Genlisea . Transfer cells are probably involved in absorptive or secretory activity. The sessile glands and the filament head appear to be involved with absorption of the digestive products. The foliar trichomes of D. capensis are highly modified to effectively attract and capture insects, and to digest and absorb nutrients.
P0667 – ePoster
Granules in the cells of maize root tips
Saito, S 1 , Niki, T 1 , Takahashi, M 2 , Gladish, DK 3
1 Dept of Cell Technology, Takushoku University,
2 Hachioji, Japan; Dept of Chemical Science and
Engineering, Tokyo National College of Technology,
Hachioji, Japan;
Hamilton OH, USA
3 Dept of Botany, Miami University,
A plant root, especially its tip, has an important role in biosynthesis of growth regulators (plant hormones).
However, it has not been clearly shown which tissue and/or cells in root tips are responsible for such biosyntheses. Toriyama (1978a,b and 1990) reported two categories of cells he called
α
-cells and
β
-cells in the root tips of Pisum sativum , Phaseolous aureus and Vicia faba , and he showed that
α
-cells (in xylem) stained strongly and
β
-cells (in phloem) stained weakly with methylene blue. He also reported that large granules were present in
β
-cells. The present authors studied whether the granules that Toriyama observed in these dicots appeared also in monocot roots, and considered what roles they might have in cell division, cell differentiation and root growth from the point of view of plant hormone synthesis.
Cultivation methods were modified after Gladish and
Niki (2000). Vermiculite in beakers was moistened with distilled water (DW). The beaker tops were covered with aluminum foil and sterilized by autoclave. Maize seeds
( Zea mays L. var. amylacea ), a monocot, were placed in the vermiculite, at constant 20˚C in continuous darkness.
4 or 5 d after planting roots were collected for light (LM) and transmission electron (TEM) microscopy. 2–3 mm long segments were excised and immersed in fixation solution. Ethanol dehydrated segments were embedded in
Technovit 7100 or Spurr resin. Sections were stained with 0.2% toluidine blue ‘O’ for LM and with uranyl
550 acetate and lead for TEM. Granules were visualized by
LM in the cells of maize root tip. Each granule appeared to be made up of uniformly high-density material and located in the small vacuoles of root tip cells. The size of the granules was estimated to be about 0.5–2
µ m.
Longitudinal sections of roots showed that the maximum distribution of granules appeared in cells located 800 to
1800
µ m from root tip. Transverse sections showed that these granules were observed first in cells around proto- and metaxylem elements, and later in pericycle, endodermis and the inner layer of cortical cells, depending on distance from the root tip. TEM observation showed the granules formed in the cytoplasm of cells, were then transferred into a vacuole by invagination, and consequently are membrane-bound.
Cytoplasmic aggregations of polyribosomes were associated with these events, and polyribosomes appeared to make up the bulk of granule contents. In our studies with maize roots, the distribution of granules was not restricted to
β
-cells (phloem). Granules were produced in the cytoplasms of root tip cells and ultimately contained by a membrane in vacuoles, unlike other inclusions, which suggests a function other than storage. The appearance of granules in specific tissues at different times, suggests that granules are associated with particular developmental events perhaps associated with hormone synthesis. The nature of the contents or how granules function remain an open question.
P0669 – ePoster
Phenotypic and molecular characterization of temperature-sensitive mutants of
Arabidopsis
that form fasciated roots under the restrictive temperature
Ohtsuka, K 1 , Konishi, M 2 , Kinoshita, A 3 , Sugiyama, M 1
1 Botanical Gardens, Graduate School of Science,
University of Tokyo, Japan; 2 Dept of Applied Biological
Chemistry, Graduate School of Agricultural and Life
Sciences, University of Tokyo, Japan; 3 Dept of Biological
Sciences, Graduate School of Science, University of
Tokyo, Japan
We isolated many temperature-sensitive mutants of
Arabidopsis by screening of an EMS-mutagenized population with adventitious root formation as an index phenotype. Out of them, rrd1, rrd2, and rid4 are characterized by forming fasciated lateral roots at the restrictive temperature. Temperature-shift experiments with a semi-synchronized lateral rooting system showed that lateral root primordia of these mutants develop into fasciated roots when exposed to the restrictive temperature at the initial stage. Detailed observation of arising primordia indicated that extra formative cell division in the pericycle leads to the fasciation phenotype. In rrd1, rrd2, and rid4, besides root development, several aspects of development and growth were found to be also temperature-sensitive. Of particular note, embryogenesis was severely interfered with by each of the three mutations at the restrictive temperature.
Double mutant analysis showed that any combinations of the mutations strikingly aggravate the embryo phenotype and eventually cause embryonic lethality even at the permissive temperature, suggesting close functional relations among RRD1, RRD2, and RID4. Positional cloning revealed that the RRD1 and RID4 genes encode

a poly(A)-specific ribonuclease (PARN)-like protein and a pentatricopeptide repeat (PPR) protein, respectively.
Sequence analysis of the rrd2 genome detected a missense mutation in a gene encoding another PPR protein, to which the temperature sensitivity of rrd2 might be attributable. In consideration of the known functions of PARNs and PPR family proteins, it is possible to speculate that the RRD1, RRD2, and RID4 proteins cooperate in some RNA processing event. This possibility is currently under examination. In the present paper, based on all the results obtained, we will discuss possible molecular functions and developmental roles of
RRD1, RRD2, and RID4.
P0672 – ePoster
Morphogenesis of fruit in palm family
Romanov, M 1 , Bobrov, A 2
1
2
Main Botanical Garden Nm. Tsitsin NV RAS, Russia;
Dept of Biogeography, Geographical Faculty, MV
Lomonosov Moscow State University, Russia
The system of Arecaceae was recently reformed based on the results of molecular data, so morphological plesiomorphies should be found for taxa of different range. Pericarp structure is of particular importance for this and we try to reconstruct the scheme of pericarp anatomical structure transformation in palms. Basal differentiation of pericarp was described for fruits of
Eugeissona , Coryphoideae–Borasseae and Nypa . It is represented by single-layered exocarp and endocarp; mesocarp is extremely multylaered. The third zone of mesocarp comprises lignified stone of the carpel, and the forth (innermost) zone is parenchymatous and partly compressed in mature fruit. The fruits of Eugeissona and
Coryphoideae–Borasseae are described as a new morphogenetic fruit type – the pyremarium of Lataniatype (Bobrov, Melikian, Romanov 2009). The structure of Coryphoideae–Trachycarpeae fruits is treated as derived from Nypa type of pericarp differentiation, but they are more specialized and the mode of stone development is different (Bobrov, Romanov, Melikian
2008; Murray 1973). This morphogenetic fruit type of apocarpous fruits is called the drupe of Rhapis-type, which specialized subtype is found in Trachycarpeae and basal subtype – in Nypa . Further specialization of pericarp of the drupe of Rhapis-type brought to origin of apocarpous berry of Coryphoideae: Schippia, Itaya,
Zombia, Cryosophila, Chuniophoenix (and Phoenix ,
Biradar & Mahabale, 1969) the fruit type with fully specialized parenchymatous mesocarp. The anatomy of the Sabal pericarp is similar, but the fruit is hemisyncarpous berry. The fruits of all Calamoideae
(excluding Eugeissona ) are paracarpous berries, which are covered with scales, making the structure of pericarp more complex. The sclerenchymatization of the inner zone of mesocarp and endocarp in pyrenaria of Lataniatype results in origin of syncarpous fruits with the stone composed by the inner zone of mesocarp and the endocarp, the pyrenarium of Butia-type. Histogenesis of pericarp in this fruit type was described by S. Murray
(1963) in Syagrus s. l. and studied by us in 15 more genera of Cocoseae. Pyrenarium of Butia-type is basal fruit type of Arecoideae: the fruits of other studied tribes of the subfamily mostly have derived pericarp. The general differentiation of pericarp is preserved in case of partial reduction of inner zone of mesocarp in
Pelagodoxa (Chapin, Essig & Pintaud, 2001), Manicaria , and some Areceae (Essig, 1977 1999; Essig, Bussard &
Hernandez 2001; Essig & Hernandez 2002; Essig &
Young 1979). It is also revealed in fruits of Phytelephas.
This pyrenarium type gradually transforms into syncarpous berry within Areceae (Essig, Manka &
Bussard, 1999; Mendonça et al., 2008; Uhl & Dransfield,
1987). Another line of fruit morphogenesis is revealed in
Ceroxyloideae, Arecoideae–Chamaedoreeae s. l. and
Coryphoideae–Caryoteae: Murray (1973) determined, that the stone in fruits of Caryota , Chamaedorea and
Hyophorbe develops ‘solely from the cells of locular epidermis’ (p. 100). This fruits are referred to pyrenarium of Ilex-type with the stone made by endocarp only.
Hemisyncarpous fruits of this type develop in
Pseudophoenix and eusyncarpous – in Ceroxylon,
Ravenea and Juania.
P0674 – ePoster
Breeding system of
Magnolia denudata
with emphasis on thermogenesis flower and anatomical changes in ovule development
Ruohan, W 1 , Zhixiang, Z 1 , Shulin, M
Yiyuan, Z 1
1 , Xiangyu, L 1 ,
1 Beijing Forestry University, China
The family Magnolia ceae, one of the most primitive among angiosperms, is mainly distributed throughout
East and Southeast Asia, southeast North America,
Central America, and South America. Magnolia denudata is an excellent ornamental and ecologically important tree that exhibits low fecundity because of seed abortion. To better understand the cause of this low sexual reproduction, pistil receptivity and pollen quality were investigated using culture medium and histochemical staining, with emphasis on pollen viability at different flowering stages. Cross-pollination, geitonogamy, and self-pollination were undertaken on flowers of sample trees. We found that flowers produce a large number of pollen grains available for effective pollination. Pollen viability varied at different stages, but was highest during stage 3, in which the stamens detach from the axis, the anthers dehisce, and the gynoecium stigma starts to fade. The duration of pistil receptivity was approximately five hours. These results indicate that poor fertilisation may occur if the optimum period for pollination is missed or the stigma receives poor-quality pollen grains from stages other than stage 3. Although dichogamy avoids selfing, it has led to considerable geitonogamy. Our anatomical analysis of ovule and seed development further revealed that fertilisation occurred in samples with geitonogamy, but that the embryo degenerated in the torpedo stage, demonstrating that aborted seeds were produced by fertilisation, rather than by pseudogamy. Our results provide new insights into the mechanism of reduced seed set under natural conditions.
As a continue study of examining breeding system of
Magnolia during these developmental processes, it is also found that Magnoliaceae owns the typical characters of the thermogenesis flower in basal angiosperms.
Flower temperatures were high when they first opened and were female, which associated with receptivity of
551

female. High enhances the evaporation of floral scents and attract visitors in early spring.
P0675 – ePoster
Preferential imaging by cryofluorescence microscopy of bundle sheath chloroplasts in leaves of
Zea mays
Sarafis, V 1 , Falat, I 1 , Vacha, F 1
1 Institute of Physical Biology, University of South
Bohemia
In sections of living leaves of maize the chloroplasts of bundle sheaths can be readily visualised by the emitted fluorescence of the chloroplasts. Placing the leaf sections at liquid nitrogen temperatures cuases strong enhancemnet of photosystem I regions of chloroplasts.
Bundle sheath chloroplasts are strongly fluorescent due to their photosystem I content , a signal which is considerably reduced at room temperature, and because photosystem I contains regions of other mesophyll chloroplasts.
P0676 – ePoster
An in vivo viewing by super resolving, fluorescence structured illumination microscopy of
Dawsonia superba
dry spores
Sarafis, V 1 , King, E 2
1 Institute of Physical Biology, University of South
Bohemia, Czech Republic; 2 Dundee University, UK
Dry spores of Dawsonia superba were imaged as fluorescing structures for both walls and chloroplasts using excitation with blue and green laser light and viewed using structured ilumination microscopy.using the fluorscence emitted. A resolution gain of double that of the best oil immersion lenses used was obtained.
P0677 – ePoster
Localization of an aquaporin TIP2;2 in phytochrome
1
A-deficient mutant
Sato-Nara, K 1 , Hashimoto, K 2
2
Faculty of Science, Nara Women's University, Japan;
Graduate School of Humans and Sciences, Nara
Women's University, Japan
Aquaporins are located in biological membranes and involved in water transport across them. Transcriptional and post-translational regulations of aquaporins are quite important to modulate water transport through the plant body. Light is one of the environmental factors to influence aquaporin expression. In Arabidopsis thaliana , an aquaporin gene, TIP2;2, is up-regulated by dark adaptation. Transgenic plants (background: Col-0) expressing a TIP2;2-GFP fusion protein driven by its own promoter, TIP2;2-GFP signals in the dark-grown plants were extremely brighter than in the light-grown plants. In the root epidermal cells of the dark-grown
TIP2;2-GFP/Col-0 plants, TIP2;2-GFP signals were localized in complicated membrane structures as well as tonoplast. Because phytochrome A seems to regulate
552
TIP2;2 expression, we also generated the TIP2;2-GFP transgenic plants of which background lines were Ler, phyA-201, and phyA-211. We compared TIP2;2-GFP signals in these transgenic plants, and found that the signals in TIP2;2-GFP/phyA-201 lines were brighter than
TIP2;2-GFP/Ler lines. Furthermore, in the epidermal and cortex cells of light-grown plants, TIP2;2-GFP signals were localized in approximately simple membrane structures (probably tonoplast) in TIP2;2-GFP/Ler lines, while in both simple and complicated membrane structures in TIP2;2-GFP/phyA-201 lines. These complicated structures in TIP2;2-GFP/phyA-201 were similar to those observed in dark-adapted TIP2;2-
GFP/Col-0.
P0678 – ePoster
Identification of blue and red
Vanda
with SEM Of aerial velamen roots
Selvam, ABD 1
1 Botanical Survey of India
Orchids are important economic plants. The ornamental orchids are traded either as potted plants or as cut flowers. Based on the floral colours, the two orchid species Vanda coerulea Griff. ex Lindl. and Renanthera imschootiana Rolfe have been named as Blue Vanda and
Red Vanda respectively. Both the species are endangered in the wild and are enlisted in the Appendix II of CITES
(Convention on International Trade in Endangered species of Wild Fauna and Flora). Endorsing the CITES decisions, the Government of India has placed these two plant species in the Negative list of Exports, which bans their collection from wild sources. The Blue Vanda and
Red Vanda could be easily identified when they are in flowering. However, it is difficult to identify them in their vegetative condition since both the species resemble one another superficially. In the present study, efforts have been made to identify them in their vegetative condition by studying the anatomical structures of
Velamen roots under SEM (Scanning Electron
Microscope). Light microscopic studies have also been carried out to supplement the diagnostic process of the studied taxa. The presence of medullary vascular strands/bundles in Red Vanda seems to be a unique diagnostic feature. Whether this kind of medullary vascular bundles occur in other species of Renanthera and Vanda or in other members of Orchidaceae is to be investigated.
P0679 – ePoster
Physiological analysis of seed development and germination in seagrass (
Zostera marina
)
Shiota, H 1
Tanaka, I 1
, Bando, Y 1 , Yanada, K 1 , Kurokawa, K 1 ,
1 Graduate School of Nanobioscience, Yokohama City
University, Yokohama, Japan
Zostera marina is a kind of marine angiosperms
(seagrasses), and is widely distributed in the Northern
Hemisphere. They live under the shallow sea throughout the life cycle. Seeds of Z. marina develop and germinate

in seawater. Here, we analyzed physiological phenomena during seed development and germination in Z. marina from Tokyo Bay, Japan. First, we determined the stages of seed and fruit development according to the long axis length, fresh and dry weight, and water content of developing seeds and fruits. The maximum values of the long axis length and dry weight were observed at 28 days after flowering (DAF), and then seeds were released from the fruit tissues at 35 DAF. Next, we analyzed the germination properties of mature seeds (after 80 DAF).
Seed germination was promoted at low temperature
(15°C), at low salinity (below 22.5‰), and in the dark.
Higher rate of seed germination was also observed at lower osmotic pressure. During seed germination, the coleoptile elongation was accelerated under anaerobic condition (below 2 mg/l dissolved oxygen, DO.), whereas the first leaf formation was accelerated under aerobic condition (7 mg/l DO.). Thus, the environmental factors strongly affected the seed germination, coleoptile elongation, and first leaf formation in Z. marina . In contrast, the effects of phytohormones on the phenomena were weak. Our results suggest that Z. marina has unique physiological features during seed development and germination to adapt to the marine environment.
P0680 – ePoster
Flower anatomy and evolution in species of
Rauvolfioideae (Apocynaceae): preliminary results
Rio, MCS 1 , Santos, MCJ 2 , Simões, AO 3
1 Universidade Estadual de Campinas, Departamento de
2
Genética, Evolução e Bioagentes, Campinas, Brasil;
Universidade Estadual de Campinas, Departamento de
Botânica, Brasil; 3 Universidade de São Paulo, Escola de
Artes, Ciências e Humanidades, Brasil
Rauvolfioideae is a paraphyletic assemblage of taxa encompassing the basalmost lineages within
Apocynaceae. The subfamily has been traditionally characterized as having flowers with a simple morphology, in which anthers and style-head are undifferentiated and do not form a gynostegium. This view has been challenged by a number of morphological, anatomical and phylogenetic studies, which showed that floral traits are highly variable within Rauvolfioideae and originated by complex evolutionary pathways. Nondifferentiated style-heads, for example, may have arisen independently several times by irreversible changes from ancestors bearing complex flowers. Our aim is to study the floral anatomy and morphology of selected taxa from
Rauvolfioideae, in order to test homology hypotheses concerning the evolution of floral organs. So far, flowers of five species belonging to four tribes have been analyzed: Aspidosperma cylindrocarpon
(Aspidospermeae), Carissa carandas (Carisseae),
Lacmellea arborescens , L. gracilis (Willughbeieae), and
Rhigospira quadrangularis (Tabernaemontaneae). The most complex flowers were observed in R. quadrangularis . In this species, anthers have lignified cells at the adaxial side forming massive wings. The style-head main body is differentiated into a 5-lobed upper portion, a massive enlarged part covered by secretory cells and a basal ring formed by epidermal cells directed downwards. The flowers from the other four species have a more simple morphology. Anthers are non-lignified and the style-head is undifferentiated, with two conspicuous, non-secretory apical appendages and a main body completely covered by secretory cells without projections or lobes. Despite their overall similarities, flowers from these four species have significant differences in terms of anther size, style-head body shape and distribution of idioblasts in the style-head apical appendages. In A. cylindrocarpon , the ovary is apocarpous and half-superior, with a nectariferous tissue in the form of a conspicuous thickening at the base.
These two features are unusual in the genus and have been previously reported only in one other species, A. quebracho-blanco. In C. carandas , L. arborescens and L. gracilis , the ovary is syncarpous and superior, without a discernible ring or thickening at the base, but evidence of a nectariferous tissue was detected. Calycine colleters were observed in L. arborescens , L. gracilis , and R. quadrangularis . Colleters of R. quadrangularis are conspicuous and divided into a large secretory head on the top of a short stalk. On the other hand, L. arborescens and L. gracilis have smaller colleters, not differenciated into head and stalk. Our results, though preliminary, support the idea of complex evolutionary pathways in the flowers of Rauvolfioideae. These pathways can be better understood by detailed anatomical studies that will contribute to ongoing phylogenetic analyses and ancestral character state reconstructions.
P0682 – ePoster
1
The adaptive radiation of the leaf venation in
Hawaiian C
4
Euphorbia
: 'shedding' veins in shade reduces cost
Sporck, MJ 1 , Sack, L 2
University of Hawaii, USA; 2 University of California,
USA
The diversity of leaf venation architecture within and across lineages is gaining increasing interest as a source of functional adaptation to contrasting environments. The
C
4
Hawaiian Euphorbia (Euphorbiaceae) radiated from one colonizing species into nearly 30 taxa. This group includes a variety of life forms, from creeping woody sub-shrubs to trees over 6 m tall, with taxa adapted to diverse habitats, from rain forest to dry forest to coastal strand. The leaves of the taxa in this group differ strongly, with 80-fold variation in leaf size and 8-fold variation in leaf mass per area. One study (Herbst 1971), pointed out a qualitative trait unique to this group,
'disjunct minor veins,' unattached to the rest of the vein network and surrounded by mesophyll cells. No study has quantified the venation architecture or its relationship to environment for this key radiation. For 27 native
Euphorbia taxa, we cleared leaves chemically and quantified 40 traits relating to venation architecture, including densities of all vein orders (i.e., length/area) and of disjunct veins. We tested for correlation of venation traits with climate and habitat, and with other aspects of leaf structure and composition. We hypothesized that leaves of taxa distributed at higher temperatures and establishing at higher irradiance would have greater vein density, and that venation architecture would depend on leaf size. We hypothesized that disjunct vein formation would be associated with moist rainforest habitats, as in shaded habitat these C
4
species might not
553

suffer from the loss of vein length and even might benefit from reduced construction cost. We found strong support for these hypotheses. Hawaii's isolated location and strong climatic gradients have led to strong diversification and apparent adaptation of venation characteristics, providing a model for understanding these traits in other lineages.
P0683 – ePoster
The nectaries and nectar secretion in the flowers of
Fritillaria meleagris
L. (Liliaceae) – an endangered plant in Poland
Stpiczyñska, MG 1 , Zych, M 1 , Nepi, M 2
1 University of Warsaw, Botanic Garden, Warszawa,
Poland; 2 University of Siena, Dept of Environmental
Sciences, Siena, Italy
In Poland, Fritillaria meleagris is critically endangered plant with only 2 natural population noted. The studies on nectar secretion and pollination in F. meleagris were conducted in 2006–2010, in nature reserve ‘Szachownica w Krównikach’, SE Poland, on an area of approx. 20 ha.
In this reserve, Fritillary plants emerge flowers about middle April and bloom for the next 7–8 days. Nectar is secreted in perigonal nectaries located adaxially, at the base each of six tepals. They have a form of groove 10–
12 mm long and 2–3 mm wide. The nectary is composed of one layer of secretory epithelial cells and 2–3 layers of subsecretory parenchyma. It is supplied with several vascular bundless. Cell walls of the nectary develope prominent ingrowths concomitantly with secretory activity, and in the cytoplasm numerous secretory vesicles are present. Traces of secretion appears one day before flower opening and nectar is secreted until the end of anthesis. Maximum amount of nectar secreted (29.9 mg, on average) was noted in the stage of full anthesis, and also at this stage stigma showed the strongest receptivity. Concentration of nectar ranged between
25.2% just after flower opening to 70% at the maximum stage of anthesis. The nectar is composed of glucose, fructose and sucrose in almost equal proportions, and it is reach in amino-acids. The difference in volume and concentration of nectar secreted in the nectaries located on the tepals of outer and inner whorl was insignificant.
The flowers are important source of pollen and nectar for local bumblebee queens that are key pollinators of F. meleagris .
P0684 – ePoster
The dynamics of nucleolar small G protein AtNog1-1 in plant nucleus is cell cycle and cell development dependent
Suwastika, IN 1
Bahk, JD 2
, Im, CH
, Takeyasu, K 4 ,
2 , Bang, WY 2
Shiina, T 5
, Ohniwa, RL 3 ,
1 Tadulako University, Japan;
University, Republic of Korea; 3
2 Gyeongsang National
University of Tsukuba,
Japan; 4 Kyoto University; Jap[an;
University, Japan
5 Kyoto Prefectural
NOG1 is a nuclear GTP-binding protein which is conserved among archaea and eukaryotes (including
554 animals, plants and fungi). It has been shown that NOG1 homologues are involved in early steps of 60S ribosomal biogenesis in nucleus of Trypanosoma , yeast and mouse.
Here we characterized the plant Nog1 protein in
Arabidopsis . Arabidopsis has three NOG1 homologues, named AtNOG1-1, AtNOG1-2 and AtNOG1-3, but only
NOG1-1 is constitutively expressed in all the tissues examined. Actually AtNOG1-1 seems to be essential for plant development. AtNOG1-1 is directed to nucleolus and nucleoplasm. FRAP analysis revealed that the distribution of NOG1-1 protein between nucleolus and nucleoplasm was sensitive to transcription and translation inhibitors, also carbon and nitrogen starvation, suggesting its role in early steps of ribosomal biogenesis. NOG1-1 shows a dynamic behavior during mitosis. The NOG1-1 protein disappeared in premetaphase, and is rapidly re-accumulated in peripheral chromosomal region in early anaphase. We also found a novel and plant-specific sub nucleolar structure named
'nog1 body' which appears in nucleus region during late anaphase and early telophase.
P0686 – ePoster
Isolation of salt glands from
Avicennia officinalis
for microscopic studies
Tan, WK 1,2 , Lin, Q 1 , Lim, TM 1 , Kumar, P 1 , Loh, CS 2
1 Dept of Biological Sciences, National University of
Singapore, Singapore; 2 NUS Environmental Research
Institute, National University of Singapore, Singapore
Salt glands are unique specialized epidermal structures found on the aerial surfaces of some halophytic species.
Plants with salt glands regulate internal salt levels by secreting the excess salts via these glands. We focus on the study of the salt glands of Avicennia officinalis , a salt-secreting mangrove tree species surviving well in intertidal zones of the tropical and subtropical areas. In our attempts to study the salt glands at the glandular level in greater detail, we developed a method to isolate large numbers of salt glands from the leaves of A. officinalis .
By removing the abaxial epidermal cell layers from the leaves and incubating the resulting leaf tissues in an enzyme solution, salt gland-enriched adaxial epidermal peels can be detached from the mesophyll-palisade tissues after an hour of enzymatic digestion. The salt glands can then be released through the grinding of these epidermal peels. Using this method, approximately 2 ×
104 – 3.5 × 104 salt glands could be obtained from about
5 g of fresh leaf tissues. This approach freed the salt glands from the interference of neighbouring leaf cells for easy manipulations. Successful isolation of salt glands achieved within 4 hours enabled observations of salt glands to be performed within the same day. The potential of applying such a system for subsequent imaging and cytological studies will be discussed.
P0687 – ePoster
Morphogenetic gradients in silver firs (
Abies alba
Mill.) and their plasticity
Taugourdeau, O 1 , Sabatier, S 2 , Caraglio, Y 2
Barthélémy, D 2
,

1 Université Montpellier, UMR AMAP, Montpellier,
France; 2 CIRAD, UMR AMAP, Montpellier, France
Silver fir presents a great hierarchised architecture with a strict monopodial development (Massart’s architectural model). Moreover silver fir presents neither polycyclism nor immediate branching (often involved in the plastic response of primary growth). Despite these limitations, silver firs present a large range of distribution: from
Central Europe to Mediterranean mountains. The aim of this poster is to test the hypothesis that silver firs offset this limitation with a strong plasticity at the annual shoot scale and on needle morphology. The study was done in
Mont Ventoux, a French Mediterranean mountain which provides contrasted forest environments in terms of elevation and with a marked summer drought.
Quantitative traits of annual shoots of 2 years old branches (length, diameter and number of needles, dry mass) and needles (surface and dry mass) where recorded on 80 saplings growing in different light environments.
Effects of tree size and architectural position of annual shoot within tree crown on measured traits were higher than any environmental fluctuation. These morphogenetic gradients are consistent with the concept of physiological age of meristems (Barthélémy &
Caraglio 2007): huge effect of tree development, branching order and position. All studied traits present a significant plasticity which may be implies in the success of silver firs in the studied site. No clear hierarchy was found between the phenotypic plasticity of annual shoot plasticity and the one of needles. We can suggest that all measured traits act simultaneously during tree development and in response to environmental fluctuations ('a variational module' sensu Wagner et al
2007).
P0688 – ePoster
1
Plant architecture: from concepts to applications
Barthélémy, D 1 , Caraglio, Y 1
Edelin, C 2 , Heuret, P 3
Rey, H 1 , Sabatier, S 1
, Charles-Dominique, T
, Meyer-Berthaud, B
, Taugourdeau, O 4
2 , Nicolini, E
CIRAD, UMR AMAP, Montpellier, France;
UMR AMAP, Montpellier, France;
ECOFOG, Kourou, France; 4
3
2
2
1
,
,
CNRS,
INRA, UMR
UM2, UMR AMAP,
Montpellier, France
Since Hallé & Oldeman (1970), plant architecture concepts are used when studying plant form and ontogeny. Using the identification of several morphological criteria and considering the plant as a whole, from germination to death, architectural analysis is essentially a detailed, multilevel, comprehensive and dynamic approach to plant development. After twenty years, this approach was completed by appropriate quantitative methods of mathematical analyses and modeling approaches (de Reffye et al. 1991). Recent researches in this field have greatly increased our understanding of plant structure and development and have led to the establishment of a real conceptual and methodological framework for plant form and structure analysis and representation (Godin & Caraglio 1998;
Guédon et al. 2001; Barczi et al. 2008; Mathieu et al.
2009). In 2007, Barthélémy & Caraglio published a large review on plant architecture which provides generic terminology and the concepts for plant architecture interpretation. The current poster aims to briefly illustrate some applications of the main concepts like architectural unit, reiteration process, morphogenetical gradients and physiological age of meristems. Specific data and results are given to provide reproducible examples on other species. Theses concepts are also useful tools for sampling plant structure in regards to ecophysiological questions on leaf physiology (Roggy et al. 2005; Coste et al. 2009; Leroy et al. 2009) and wood anatomy and hydraulic (Cochard et al. 2005). Finally, we address some present questions on phenotypic plasticity
(Stecconi et al. 2010; Charles-dominique 2010;
Taugourdeau et al, 2011), forestry (Rutishauser et al,
2010), agronomy (Rey et al. 2008) and paleobotany
(Meyer-Berthaud et al. 2010) and particularly how architectural features can become relevant original traits for ecological and evolutionary studies.
P0689 – ePoster
Anatomical studies of the gynoecium of
Anthurium
Schott (Araceae)
Temponi, LG 1 , Poli, LP 1 , Coan, AI 2
1 Unioeste, Cascavel, Brazil; 2 Unesp, Rio Claro, Brazil
Anthurium is the largest genus of Araceae with approximately 1000 species, distributed from Mexico to
Argentina, and especially in Brazil with about 120 species. The objective of this work was to study the anatomy of the gynoecium of 11 species of Anthurium , aiming to contribute distinguishing characteristics of the family. All of the species studied have: bicarpellate and bilocular gynoecium, locules filled with mucilage; cleftshaped stigma, formed by unicellular trichomes, a stylar canal lined with transmitting tissue formed of short unicellular trichomes; and a proximal stylar region and placenta formed by long secretory trichomes. The epidermis in the region adjacent to the stigma, the length of the stylar region, the width of the ovarian septum, and placentation vary in the species studied here. What is noteworthy in this study is the confirmation of the occurrence of secretory trichomes in the stylar canal and in the placenta of all the species of Anthurium studied, confirming the trait for the genus in Araceae.
P0690 – ePoster
Endopolyploidy in
Dendrobium
Chao Praya Smile
1
(Orchidaceae)
Teng Seah, K 1 , Chiang Shiong, L 1
National University of Singapore, Singapore
Dendrobium is one of the largest genera in the family
Orchidaceae. The hybrids are commercially important in the orchid industry. Different parts of Dendrobium plants have been used as explants for micropropagation.
Unfortunately, not all the plants produced through micropropagation were found to be true-to-type.
Variations have been reported among plants regenerated from explants of in vitro cultures. A possible cause of the observed somaclonal variants could be the pre-existing ploidy differences in somatic cells of the initial explants.
555

In this paper, the occurrence of endopolyploidy in greenhouse-grown Dendrobium Chao Praya Smile, a commercial hybrid, was analysed using flow cytometry.
Different tissue types harvested from different developmental stages of the plants were compared.
Nuclei of up to 8C DNA content were present in the first leaves of greenhouse-grown plants that were in their vegetative phase. On the contrary, analyses of leaves from plants that had bolted, nuclei of up to 32C DNA content were detected in the first leaves. Shoot tips and axillary buds from plants in the vegetative stage were found to contain a mean frequency of 4.6 and 8.9% 8C nuclei respectively. Tissues near the base of 6-cm root contained a mean frequency of 18.7% nuclei with 8C
DNA content, whereas 8C nuclei accounted for 9.3% of the total nuclei population in the tip analysed. Since the ploidy variations of the floral parts harvested at different days after anthesis were similar, endopolyploidy in the floral tissues was not developmentally regulated. The maximum C-value detected in the floral tissues is 16C.
P0692 – ePoster
Biochemical responses of Iranian alfalfa ecotypes to salt stress
Torabi, M 1 , Halim, R 2 , Chokan, R 3
1 Isfahan Center For Research of Agricultural Science and Natural, Iran; 2 University Putra Malaysia; and Plant improvement Institute of Iran
3 Seed
An experiment was conducted to characterize biochemical responses to salt stress of five Iranian alfalfa ecotypes. A hydroponics system was used at University
Putra Malaysia from December 2008 until April 2009.
The seeds were planted in plastic pots filled with sand and immersed in nutrient solution. After 2 months when the plants became mature and stable the EC of nutrient solution was increased by adding sodium chloride gradually. At every 15 days the NaCl was increased by 3 dS/m, and the final EC was 18 dS/m. Samples were taken at each time when plants were exposed to three levels of salinity: 6, 12, and 18 dS/m. The biochemical components measured were: root soluble sugars
(fructose, glucose, and sucrose), free proline, and total amino acids. Three soluble sugars in root including fructose, glucose, and sucrose were measured using
HPLC. The results indicated that when alfalfa is exposed to salt stress the monosaccharides were more affected than the disaccharides. There was significant linear increase of fructose in roots under salt stress for all ecotypes. The concentration of glucose in root increased under salt stress. There was significant difference between ecotypes in terms of concentration of glucose under salt stress. Influence of salt stress on concentration of sucrose was same with reducing sugars where there was significant difference between levels of salt stress in terms of concentration of sucrose. The responses of ecotypes to salt stress in terms of total soluble sugar accumulation varied between ecotypes so that ecotypes numbers 2, 5 accumulated more soluble sugar and are considered tolerant than others. There was significant difference between levels of salinity, ecotypes, and interaction between salinity and ecotypes on the level of free proline. With increasing salt stress the concentration of free proline was elevated but from moderate to high
556 concentration of salt stress the increase was somewhat slow. Release of free proline of ecotypes to salt stress was variable where number 5 and 2 were the superior than others. Through evaluation of total amino acid fifteen, amino acids were detected by HPLC detector.
The results showed that the total amino acids were affected by salt stress. The content of most of amino acids changed in with increasing of salt stress significantly and a few of them such as aspartic acid, glutamic acid, serine, and isoleusine did not change significantly. There was significant difference among the ecotypes relative to most of amino acids except three amino acids including aspartic acid, histidine, and tyrosine. When the mature plants of alfalfa were subjected to salt stress the content of three total amino acids including proline, threonine, and lysine was higher than control and with increasing salt stress the concentration of these three amino acids increased. The response of ecotypes relative to total amino acids was significantly different where ecotypes number 4 and 2 were superior to others.
P0693 – ePoster
Floret anthesis in maize
Tsou, C-H 1
P-C 2
, Fu, Y-l 1 , Tseng, C-M 1 , Yen, H-R 1 , Cheng,
1 Institute of Plant and Microbial Biology, Academia
Sinica, Taipei; 2 Dept of Electron Engineering, State
Univ. of New York at Buffalo, NY, USA
Floret anthesis in maize ( Zea mays L.) starts with lodicules expansion, glume opening, and then is followed by filament extension and anther dehiscence. The process is usually completed within 30–40 minutes. Water movement and cellular elongation but not cell proliferation has long been considered to cause the enlargement of lodicules and filaments and other movement of floral parts, but detailed mechanisms and the role of water transpiration at anther dehiscence are yet to be explored. HPLC analysis showed a sharp increase of glucose and fructose concentrations in lodicules at anthesis, which built up a high osmotic gradient and acted as a sink for directional water movement which accounted for more than 3x increasing in lodicule volume. TEM study revealed a thick cuticular covering on the surfaces of lodicules and anther.
Experiments with tritiated water (3H
2
O) on isolated anther cuticle further demonstrated that this cuticle layer was impermeable to water; hence water lost due to direct transpiration through anther surface is highly unlikely. A negative water potential was created in the anther as the result of high sugar concentration in the lodicules as well as transpiration of glume and lemma during the lodicules swelling and filaments elongation processes. Cryo-SEM results indicated that the anther locule and the interpollen spaces were free of water long before anthesis; whereas epidermis, endothecium, and connective cells remained fairly turgid till the moment of anthesis. At anthesis, after the filament has reached near- or fullyelongated state, anther wall cells began to show water lose unexceptionally from the apex downwards. While the epidermal cells at anther apex were shrinking, tensile force was generated due to the decrease of the anther circumference against the rather ridge secondary-wall-

thickenings equipped endothecial layer, which subsequently generated a tear on the epidermis and endothecium at IMS1 to form the anther pores. The removal of water from anther to the expanding lodicule/filament is the mechanism mediating anther pore opening while transpiration from anther surface is negligible. Dehydration from anther wall layers continued after pore opening, which also caused the separation of the weakest region underlying the IMS1 and between the paired microsporangia, a region with large intercellular spaces and weak cell adhesion; thus the epidermis and endothecium beneath the IMS1 was separated from the underlying connective and the two microsporangial locules became confluent into one pollen sac. The confluent is called ballooning in maize.
The ballooning caused the pollen grains rearrange from their originally tightly packed donut-shaped arrangement to became free from each other and attended their final spherical shape. Pollen grains were then able to be released and dispersed.
P0694 – ePoster
Just what is a flower? Developmental genetics, morphology and multi-level homology in
Triuridaceae and other families of Pandanales
Vergara-Silva, F 1
1 Instituto de Biología, Universidad Nacional Autónoma de México, Mexico
The reproductive structures of the Mexican mycoheterotrophic monocotyledon Lacandonia schismatica (Triuridaceae: Pandanales sensu APG III
2009) were first described as true flowers with an unusual homeotic arrangement: in contrast to typical angiosperm bisexual axes, their structural plan involved a tripartite, central androecium surrounded by a polycarpellate gynoecial whorl. Inspired by the 'ABC model' for the specification of floral organ identity, L. schismatica has been the focus of a long-term research project aiming to characterize the expression pattern of homologous 'floral homeotic' genes during young stages of reproductive meristem development. This project rested upon a straightforward, ABC model-based hypothesis –namely, that the homeotic position of male and female organs in L. schismatica flowers should be
(causally) related to a 'centripetal displacement' of the Bfunction, instantiated by changes in the expression pattern of orthologs of the Arabidopsis MADS-box genes
APETALA3 and PISTILLATA. A caveat for this hypothesis, however, was eventually introduced from an alternative, 'pseudanthial' morphological conception prompted by molecular phylogenetics results. According to those results, Triuridaceae was placed as sister group to Pandanaceae, where bonafide pseudanthia occur; if true, this contrasting structural view would invalidate the rationale of the aforementioned developmental genetics project. The author of the pseudanthial interpretation of triurid reproductive axes has later proposed a further morphological model, in which the absence of a distinct boundary between 'inflorescence' and 'flower' prescribes defining such axes as 'non-floral'. In turn, the results of a more detailed study of early morphogenesis in both L. schismatica and a closely related Mexican triurid – carried out by the research group which originally supported the euanthial molecular-genetic hypothesis – contested the pseudanthial interpretation; more recently, the same group has published the expression patterns of both B- and C-function MADS box genes in L. schismatica . With additional data at hand from transgenic complementation experiments in mutant Arabidopsis lines and biochemical assays of protein-protein interactions, these authors have concluded that the 'Bfunction shift' hypothesis is correct. After reassessing (i) previous observations of variable floral morphs in natural populations of the two known Mexican triurid species,
(ii) current estimations of reproductive character evolution in Triuridaceae, and (iii) the absence of a consensus on infraordinal phylogenetic relationships within Pandanales, here I argue that the available data on the developmental genetics of L. schismatica , though valuable, could still be interpreted under alternative perspectives. I claim that a thorough evaluation of homologies of genetic and epigenetic mechanisms has not been fully achieved yet – not even at the hypothesis formation stage – in the relevant literature, in order to differentially support diverse morphological concepts of reproductive axes in Triuridaceae, other Pandanales, and a few additional angiosperm families where unusual features also occur (e. g. Hydatellaceae). I conclude that the history of molecular research in Triuridaceae tells a cautionary tale about 'plant evo-devo' and its current theoretical framework, and relate my discussion to more general issues in the epistemology of evo-devo -in particular, to systematizations of 'hierarchical homology' at multiple levels of biological organization, which might apply to Triuridaceae’s complex case.
P0695 – ePoster
The remarkable Androecium evolution in
1
Helicteroideae (Malvaceae, Malvales) von Balthazar, M 1 , Nyffeler, R 2 , Schönenberger, J 3
Dept of Systematic and Evolutionary Botany, University of Vienna, Austria; 2 Institute of Systematic Botany,
University of Zurich, Switzerland; 3 Dept of Structural and Reproductive Botany, University of Vienna, Austria
Many representatives of Malvaceae (Malvales) have evolved peculiar anther structures; some of the most extreme forms are found in the subfamily Helicteroideae.
Helicteroideae are strongly supported as a monophyletic subfamily of Malvaceae and comprise the two distinct sister groups Helictereae and Durioneae. Helictereae have around six genera and Durioneae consists of the genera Neesia, Coelostegia, Kostermansia, Cullenia,
Boschia , and Durio . Neesia is sister to the remainder of
Durioneae, and Cullenia, Boschia , and Durio form the core of Durioneae. While Helictereae and the earlydiverging taxa of Durioneae generally have bithecate, tetrasporangiate anthers that are typical for angiosperms in general, the core group of Durioneae has evolved an amazing diversity of anther structures. The concept of typical angiosperm anther organization has been lost in the course of evolution and instead number of sporangia and thecae has increased and arrangement of these parts has become irregular. The evolution of these androecial structures is hypothesized to be related to different pollinator groups. The early-diverging genera of
557

Durioneae are pollinated by beetles, flies and other small flower-visiting insects, whereas the genera of the core
Durioneae are visited by larger animals such as mammals and birds. To homologize these remarkable androecial structures with those of related taxa, we present our study on the development and detailed structure of the flowers with particular focus on the androecium.
P0696 – ePoster
Coevolution of leaf venation pattern and anatomical types in C
4
plants
1
Voznesenskaya, EV 1 , Koteyeva, NK 1 , Edwards, GE 2
V. L. Komarov Botanical Institute of RAS, St
Petersburg, Russia; 2 School of Biological Sciences,
Washington State University, Pullman, USA
It is well known that C
4
photosynthesis accomplished by the Kranz type of anatomy develops in the tropical, arid or semiarid regions where the availability of CO
2
is limited. Great diversity in structural types was found in representatives of some eudicot families in density and distribution of vascular bundles (VB) respective to two layers of chlorenchyma, mesophyll (M) and bundle sheath (BS) characteristic of Kranz type anatomy in C
4 plants. On the basis of the study of leaf venation, among
C
4
species two different evolutionary trends in modification of leaf vein pattern and density as adaptation to drought and high temperature stress are suggested. One well-known trend involves a decrease of vein spacing and increase of vein density shown mainly for laminate leaves, which is also characteristic of the higher order of branches of C
3
species known as Zalenski law. In general most eudicot C species with broad
4 laminate leaves have the so called Atriplicoid type of anatomy with multiple simple Kranz units (constituted by the VB/s, M and BS) surrounding each vascular bundle.
In this case each vein is surrounded by 2 chlorenchyma cells (M+BS), and often there is no additional cells between adjacent VB. Development of Atriplicoid leaf anatomy with close vein spacing and high vein density was observed in species from 15 families. Changes in leaf morphology during plant adaptation to increasing aridity and solar radiation, and decreasing water availability, may involve also a decrease of leaf area and, in some xerophytic species, an increase in succulence
(Fahn and Cutler 1992; Eggli 2002). Thus, another trend in adaptation to drought and high temperature stress is characteristic of species having some degree of leaf reduction with further development of Kranz anatomy consisting of the single compound Kranz unit surrounding all veins located in the center of the leaf with different degree of water storage tissue development around veins. This was observed among C
4
species in a number of families including Asteraceae,
Chenopodiaceae and Cleomaceae. In representative species, increase of vein density is absent but two ways of venation pattern were found: 1) often leaves show a reduction in reticulate venation and terminal minor veins, and development of nearly parallel venation with rare anastomoses (in representatives of Asteraceae and
Cleomaceae); 2) vein network is represented by the reticulate venation with rare terminal minor veins (in genera Suaeda and Salsola , Chenopodiaceae). In summary, the increasing vein density is characteristic
558 only of broad-leaved C
4
species, while in species having reduced leaf size and/or water-storage tissue the vein density is significantly lower and rather similar for C and C
4
species.
3
P0697 – ePoster
The mechanism of epigyny formation in flower of
Aizoaceae, its evolutionary interpretation and significance for family phylogeny
Vovk, M 1
1 T Shevchenko Kyiv National University, Fomin
Botanical Garden, Ukraine
Within Aizoaceae one observes big variety of flower structure of which the main features are ovary type
(perigyny up to complete epigyny), placentation type
(central-angular up to basally-parietal), androecium structure (single up to numerous) and perianth structure
(simple up to complex). Such variety is systematic attribute and may be reliable criterion of family phylogeny and level of primitiveness or evolutionary advances. It's clear that during evolution for protection of generative spheres arise tendency to epigyny formation which at most specialized Ruschioideae accompanied by changing of placentae position. Mesembryanthemoideae and Ruschioideae are monophyletic subfamilies and their flowers have similar structure but differ by placentation type and gynoecium structure. Mesembryanthemoideae represents by typical syncarpous gynoecium with centralangular placentation. Flower of Ruschioideae ex facte have normal syncarpous gynoecium with basally-parietal placentation. But detail vascular analysis shown that in
Ruschioideae gynoecium strongly deform. Flower conducting system being highly conservative can keep features characteristic for initial forms and reflects some structural changes in flower during evolution. In addition traces of conducting bundles of missed structures could remain. Vascular-anatomic researches of Delosperma echinatum, Trichodiadema stelligerum, Lampranthus haworthii, Hereroa incurva, Conophytum aequale
(Ruschioideae) have shown that forming of epigyny goes by receptacles invagination to what presence of loops of receptacle bundles in ovary walls testifies. Owing to receptacles invagination occur carpels deformation which growing together with a conic receptacle by ventral seams are compelled to turn inside out together with placentae. Detailed research has shown that placentary bundles are complex formations arising owing to confluence of receptacle bundles with upper sites of carpel ventral bundles. It’s obvious that such confluence is caused by coalescence of carpel ventral seams with receptacle. In flower Mesembrianthemoideae with weak epigyny ( Aptenia cordifolia, A. lancifolia ) is observed insignificant receptacle invagination too and loops of receptacle bundles in its ovary are found. Most likely here ventral seams of carpels not adhere to receptacle and it invagination doesn’t entail carpel deformation or invagination degree is so insignificant that doesn’t touch on carpel tissue. Comparative analysis of
Mesembryanthemoideae and Ruschioideae enable to conclude that features of evolutionary specialization will be complete epigyny, deformed gynoecium with basallyparietal placentation. Mesembryanthemoideae is earlier branch of Aizoaceae in comparison with Ruschioideae which admit the most evolutionary advanced group of

Aizoaceae. Data of vascular-anatomic analysis convincingly testify to axial nature of inferior ovary of
Aizoaceae. Epigyny forming by receptacle invagination is precisely connected to placentae displacement on parietal position what is evolutionary expedient. On basis of ontogenetic and vascular-anatomic data it’s possible to construct morphogenetic line with set of intermediate forms. All these ovary forms are presented by nowadays existing species. During epigyny forming the carpel cavity appears more closed, accordingly more protected from environment influence, and connects with environment only by ventral channel which is extended during ovary deepening. Besides such seed-bud arrangement and ovary structure, also presence of false median septa, are more favourable at ripening and opening of fruits. Mechanism of epigyny formation by receptacle invagination in Aizoaceae similar to Cactaceae that grounds to consider these families closely related.
P0698 – ePoster
Arabidopsis inositol polyphosphate 6-/3-kinase
(AtIpk2
β
) regulates floral transition
1
Xia, H 1
College of Life Sciences, Wuhan University, China
In Arabidopsis , floral transition is regulated by a network requiring endogenous control and environmental signals.
A set of pathways with regulators have been discovered.
In the former study, we have reported that the
Arabidopsis inositol polyphosphate 6-/3-kinase gene
(AtIpk2
β
), which encodes a key enzyme in PI metabolism, regulates photoperiodic flowering by inhibiting CONSTANS (CO). Here we analyzed further molecular mechanisms of AtIpk2
β
in flowering regulation. We examined the expression level of AtIpk2
β in red, blue and white light, which are the primary signals accepted by photoreceptors in photoperiod pathway. The results showed that AtIpk2
β
is up-regulated in the light compared to the darkness control. Moreover, the expression of AtIpk2
β
is down-regulated in phytochrome
A (phyA) mutant, which is deficient in red/far-red light photoreceptor PHYTOCHROME A (PHYA). In addition, ipk2
β
mutant showed longer hypocotyls in continuous far-red light and a significant degree of cotyledon unfolding as phyA mutant when treated with daily pulsed red light. Further analysis revealed that the expression rhythms of TOC1 and GI generated by circadian clock under long days are changed in ipk2
β
.
Taken together, our results indicated that AtIpk2
β
may regulate the floral transition by modulating PHYA signaling pathway and circadian clock.
P0699 – ePoster
Living wood fibers act as starch storage and transport pathway in black locust (
Robinia pseudoacacia
)
Yamada, Y 1 , Awano, T 1 , Fujita, M 1 , Takabe, K 1
1 Graduate School of Agriculture, Kyoto University,
Japan
Most wood fibers complete their cell wall formation and die before the dormant period. In some species, for example some trees of Aceraceae and Fabaceae and many shrubs, however, wood fibers retain their living protoplast for months or years. These fibers have been called living wood fibers (LWFs). LWFs store numerous starch grains during the dormant period, as axial and ray parenchyma cells do. In black locust ( Robinia pseudoacacia ), almost all wood fibers in the outer part of the annual ring are LWFs. In the outermost ring, starch is accumulated during the summer, retained in winter, and metabolized during spring. We determined the starch content of LWFs, ray parenchyma, and axial parenchyma using image analysis further than chemical analysis.
More than 70% of the starch grains in the outermost ring were stored in LWFs during winter. In LWFs, the starch decrease started prior to cambium reactivation and advanced gradually at leafing and flowering. After the breakdown of starch in spring, LWFs resulted in cell death. From detail observation of cross and tangential serial sections, we calculated LWF: fs contact ratio to parenchyma and examined the contact ratio and existence of starch and nuclei during starch degradation period.
LWF: fs contact ratio to parenchyma was low. Noncontact LWFs tended to lose starch grains and nuclei earlier than contact LWFs. These results indicated that there is the transport pathway from non-contact LWFs to parenchyma via contact LWFs. Therefore, LWFs themselves act as transport pathway. These results indicate that LWFs in black locust function as single-use large-capacity starch storage and pathway for sugar transport.
P0700 – ePoster
Phyllotactic transitions in
Diphasiastrum digitatum
Yin, X 1 , Meicenheimer, R 1
1 Dept of Botany, Miami University, USA
There is a plethora of both theoretical and empirical studies on the phyllotaxis of plants bearing megaphylls, but similar studies on plants bearing microphylls are quite rare. At present, it is unclear whether the same pattern formation hypotheses that appear to explain megaphyllous phyllotaxis can even be applied to those taxa bearing microphylls. There is some evidence that the prevalence of Fibonacci patterns observed in megaphyllous taxa is not reflected in at least one microphyllous species. If this observation is found to be more wide spread, this would suggest that a different mechanism governs microphyllous phyllotaxis. We undertook this study to further explore this possibility.
Diphasiastrum digitatum (ground cedar) is a clubmoss which bears microphylls. The arrangement of these microphylls (phyllotaxis) has never been studied at the level of the shoot apical meristem (SAM). Preliminary analysis indicated that D. digitatum has three types of
SAMs: rhizomatous, vegetative, and reproductive. The rhizome runs under ground horizontally and produces non-photosynthetic microphylls. It gives rise to vertically-oriented vegetative shoots in a pseudodistichous manner, where one vegetative shoot is initiated from the left upper portion of the rhizome and the next is initiated from the right upper portion of the rhizome in a repeating pattern. These vegetative shoots produce photosynthetic microphylls and a subset of these axes give rise to the reproductive shoots (strobili) in
559

groups of four. The distal and proximal regions of the rhizome axes exhibit different phyllotactic patterns which suggest that phyllotactic transitions occur along these stems. In general, there are two types of phyllotactic transitions: continuous and discontinuous. Continuous transitions occur via the symmetrical expansion or contraction of the circumference of the SAM's, resulting in uniform changes in the global relationships of the primordia. Discontinuous transitions result in additional or fewer primordia/parastichy being inserted or deleted around the SAM circumference. The vegetative shoots exhibit decussate phyllotaxis. It also has an isotropic dichotomous branching pattern, from which two equal daughter SAM's are produced. The reproductive shoots
(strobili) exhibit spiral phyllotaxis of sporophyll arrangement. For rhizomatous and vegetative meristems, phyllotactic parameters such as: divergence angle (the angle formed by successive primordia measured from the center of the shoot apex), plastochron ratio (the ratio of distances from a primordium to the center of the shoot apex for successive primordia), and van Iterson's parameter b (the ratio of the diameter of the smallest primordium to the circumference of the shoot apex) will be examined using scanning electron microscopy and light microscopy and the details will be reported. These parameters have been studied both empirically and theoretically to construct phyllotactic models for plants bearing megaphylls. A similar approach toward analyzing the microphyllous phyllotaxis of D. digitatum should be enlightening with regard to whether or not the theoretically models that appear appropriate for megaphyllous systems can be applied to microphyllous systems.
P0701 – ePoster
Analysis of phyllotactic pattern transitions in seedlings of
Thuja occidentalis
L.
Yin, X 1 , Lacroix, C 1 , Barab, D 2
1 Dept of Biology, University of Prince Edward Island,
Canada; current address: Miami University, USA;
2 Institut de Récherche en Biologie Végétale, Jardin
Botanique de Montréal, Université de Montréal, Canada
Phyllotaxis, the arrangement of plant organs on a shoot, raises some of the deepest questions in plant development and plant morphogenesis. During the ontogeny of some plants, the phyllotactic pattern can undergo transitions. There are two types of transitions: continuous and discontinuous. Continuous transition occurs via the symmetrical expansion or contraction of the circumference of the shoot apical meristem (SAM), resulting in uniform changes in the global relationships between primordia. The phyllotactic pattern changes from (m, n) to (n, m + n) or (n – m, n) (m < n) where m and n represent the number of the conspicuous parastichy pair (spirals joining a leaf to its nearest neighbor on the left or right). Discontinuous transition occurs via the asymmetrical expansion or contraction in localized sectors of the circumference of the SAM, resulting in additional or fewer primordia/parastichy inserted or deleted. The phyllotactic pattern changes from (m, n) to
(m, n ± 1) or from (m, n) to (m ± 1, n) (m < n). The main goal of this study is to examine different phyllotactic patterns and pattern transitions in seedlings of Thuja
560 occidentalis (eastern white cedar). Four phyllotactic patterns were observed on the main stem in seedlings of
T. occidentalis : tetracussate, tricussate, spiral, and decussate. The decussate pattern appeared to be the most stable pattern on the main stem in seedlings of T. occidentalis , since all seedlings ultimately reached this pattern. Only one phyllotactic pattern was observed on the side branches in seedlings of T. occidentalis : decussate. Four types of phyllotactic pattern transitions were observed on the main stem in seedlings of T. occidentalis : tetracussate to decussate, tetracussate to tricussate, tricussate to spiral, and spiral to decussate, all of which are discontinuous transitions and are rare. For each phyllotactic pattern, the following phyllotactic parameters were examined using light microscopy: divergence angle (the angle formed by successive primordia measured from the center of the SAM), plastochrone ratio (the ratio of distances from a primordium to the center of the SAM for successive primordia), leaf insertion angle (sector covered by a primordium), parameter §¤ (the ratio of the diameter of a primordium and the radius of the SAM), and apical angle of the SAM (the angle of the vertex of the cone used to approximate the SAM). Even though they varied widely, these phyllotactic parameters measured in seedlings of T. occidentalis fell within ranges observed in other plants for specific phyllotactic patterns. The results indicate that it is not possible to discriminate between the four phyllotactic patterns observed in seedlings of T. occidentalis by using these phyllotactic parameters
(except divergence angle, which is characteristic of a specific phyllotactic pattern). In contrast to continuous transitions, where there is a good correlation between phyllotactic parameters, there was no correlation between the phyllotactic pattern and these phyllotactic parameters in the discontinuous transitions observed in seedlings of
T. occidentalis .
P0702 – Poster
Elucidating the mechanism of poricide anther dehiscence in Melastomataceae
Cortez, PA 1 , Caetano, APS
1 , Teixeira, SP
1 , Simão, DG
3
2 , Carmello-
1
Guerreiro, SM
Universidade Estadual De Campinas, UNICAMP,
2 Universidade Federal de Uberlândia, UFU, Brazil;
Brazil; 3 Universidade de São Paulo, USP-RP, Brazil
Melastomataceae species have porate anthers; however, unlike Solanaceae and many monocots, in which the pore dehiscence process depends on the presence of a mechanical layer, often the endothecium, most of the
Melastomataceae has no evident specialized layer related to the pore aperture. Moreover, the anther structure is interesting since in Melastomataceae stamen features are important to both phylogenetic and ecological studies due to the morphology variation and to the many species with a great deal of unviable pollen. The goal of this study was to characterize the tissues that form the apical pore of the anther in several Melastomataceae from the
Miconia and Leandra genus, which may help us to understand the unknown mechanism of anther dehiscence in these megadiverse genera. The anthers were fixed in a modified Karnovsky solution and studied under light and scanning electron microscopes according to routine

protocols. Transversal and longitudinal sections obtained were submitted to coloration tests to detect the presence of cuticle in the pore region. Miconia species were the only ones with the stamens in a two-verticile fashion due to the dislocation of the larger anthers towards the external side. The anthers of both verticile had apical pores. Before the anthesis of the flowers, the apical pores of all of the species were closed by a unisseriate epidermis, the cells of which lack a cuticle. In contrast, the epidermis of the rest of the anther was covered by a thick, ornamented cuticle. Only in the endothecium of
Leandra melastomoides were some crystals of the druse type observed, but its relation to the aperture mechanism is not known. Among Myrtales families, the
Melastomataceae seems to form a clade with
Alzateaceae, Rhynchocalycaceae, Penaeaceae and
Oliniaceae, almost all of which have anthers with endothecium lacking fibrous thickening. In these families, the endothecium may or may not be present in the mature anther, with degenerating cells in the latter case. In these families, the anther aperture is not dependent to the presence of the endothecium as the mechanical layer, and the process of anther dehiscence is still unknown. However, in the Miconia and Leandra species studied here, the presence of a cuticle may be a way of preventing tissue dehydration, and the pore aperture seems to be due to the passive process of dehydration taking place only in the pore region due to the absence of the cuticle. This is the first report of the process of anther dehiscence independent of a mechanical tissue among Angiosperms. Financial support: Fapesp (2007/52030-0 and 2008/10793-0)
P0703 – Poster
Expression of SHI family genes from
Brassica rapa
L. ssp. pekinensis
effects on plant growth and development in
Arabidopsis
Lee, Y-H 1 , Hong, JK 1 , Kim, JS 1 , Kim, JA 1 , Lee, SI 1
1 National Academy of Agricultural Science, Suwon,
Republic of Korea
SHI (short internodes), putative transcription factor, is a negative regulator of gibberellin-induced cell elongation.
Extensive searches in the Brassica rapa genome allowed for the prediction of at least six different SHI-related genes on six chromosomes in the genome. Genome structural examination revealed that these genes have one intron each in their corresponding ORFs. Structure comparisons of all BrSRS ( B. rapa SHI-related sequence) proteins revealed broad conservation of the
RING finger-like zinc finger and IGGH motifs.
According to the phylogenetic relationship based on deduced amino acid sequences, the six BrSRS proteins were most closely related to Arabidopsis SRS (AtSRS) proteins; however, BrSRS proteins were dispersed in the phylogenetic tree. Semi-quantitative RT-PCR analysis indicated that the six BrSRS genes exhibited different expression patterns in various tissues and responded differently to growth phytohormones. Among these genes, BrSTY1, BrSRS7, and BrLRP1 over-expressing
Arabidopsis exhibited dwarfed plant growth and upward curling leaf. Also, these transgenic plants had narrow widths and short petioles in leaves, and showed shorter siliques and low fertility. However, transgenic plants over-expressing BrSTY2a, BrSTY2b, and BrSRS5 genes were phenotypically similar to the wild-type plants.
Based on these data, we suggest that BrSRS genes may participate in the control of developmental process of B. rapa .
P0704 – Poster
Gland diversity in legume flowers
Marinho, CR 1 , Souza, CD 1 , Barros, TC 1 , Teixeira, SP 2
1 Faculty of Philosophy, Sciences and Letters of Ribeirão
Preto, University of São Paulo, Brazil; 2 Faculty of
Pharmaceutical Sciences of Ribeirão Preto, University of
São Paulo, Brazil
Despite the fact that Leguminosae is a rich-species family, exhibiting great morphological variation, their representatives have been poorly studied in terms of floral gland diversity. Glands are structures responsible for the production, accumulation and release of special plant metabolites. They can be located in different flower parts and are often associated with pollinator attraction or plant defense. This study aimed to fill the gap in our knowledge regarding gland diversity (nectary except) present in flowers of leguminous species, enabling functional and phylogenetic inferences. Legume species were chosen in order to cover the Leguminosae subfamilies: Caesalpiniodeae ( Bauhinia rufa, Hymenaea courbaril, Caesalpinia pulcherrima, Poincianella pluviosa, Erythrostemon gilliesii ), Papilionoideae
( Platycyamus regnellii, Mucuna urens, Tipuana tipu,
Pterodon pubescens ) and Mimosoideae ( Inga edulis,
Parkia pendula, Stryphnodendron adstringens ).
Preanthesis and anthesis flowers were processed for observation under light and scanning electron microscopes. B. rufa has secretory epidermis and mesophyll in sepals and petals and hollow trichomes in sepals (abaxial surface), connective and ovary. Such trichomes have a short stalk and a pluricellular hollow head composed of one lenticular cell layer surrounding the lumen. H. courbaril sepals and petals have secretory epidermis and mesophyll and resiniferous cavities, which also occur in the ovary. C. pulcherrima petals have secretory papillose epidermis and oil idioblasts in the mesophyll. P. pluviosa petals have secretory epidermis and glandular emergences (abaxial surface), composed of a long stalk and a compact pluricellular head. Similarly,
E. gilliesii has glandular emergences in sepals (abaxial surface) and secretory epidermis in petals. Theses emergences have a long stalk and a pluricellular head with intercellular spaces. M. urens has phenolic idioblasts adjacent to the vascular bundles in sepals and petals. P. regnellii and T. tipu petals have secretory epidermis and phenolic idioblasts adjacent to the vascular bundles. P. pubescens has secretory epidermis in petals and cavities in petal and anther apices. I. edulis petals have secretory epidermis and phenolic idioblasts. P. pendula have secretory epidermis and mesophyll in sepals and petals. S. adstringens has secretory epidermis in petals and osmophores in anthers. Several types of glands were found: secretory trichomes, cavities, idioblasts, emergences, osmophores, in addition to whole tissues specialized in other functions than secretion, such as epidermis and mesophyll. It is worthwhile emphasizing the presence of less commonly described
561

glands, such as hollow secretory trichomes in B. rufa and emergences in E. gilliesii and P. pluviosa . The presence of secretory epidermis in the sepals and/or petals of all analyzed species, in addition to a secretory mesophyll, could be related to pollinator attraction. Secretory idioblasts and cavities are known in literature as glands producing metabolites against herbivores. The presence of secretory content in theses glands during the floral anthesis raises doubts about their real function, whether attractive or defensive glands. B. rufa hollow trichomes and H. courbaril resiniferous cavities are structures shared by other representatives of their respective tribes.
P. pluviosa and E. gilliesii (Caesalpinieae tribe) share emergences. Our data show that gland diversity in legume species studied can have adaptive and taxonomic value. Funding Institution: FAPESP.
P0705 – Poster
Pericarp and seed structure in three species of
Passiflora
(Passifloraceae-subgenus
Decaloba
(DC.)
Rchb. section
Xerogona
(Raf.) Killip)
, Carmello-Guerreiro, SM 1
1
Milani, JF 1
Universidade Estadual de Campinas, Brazil
The Passifloraceae family is distributed in tropical and subtropical climates and include lianas, herbs, shrubs and small trees. The genus Passiflora L. is the largest and most representative of the family with approximately 525 species divided into four subgenera. Decaloba distinguishes itself by having small flowers and fruits when compared to other species from the subgenera
Astrophea and Passiflora . Decaloba is divided into many sections and subsections, one of which is Xerogona , including Passiflora capsularis L., P. citrina MacDougal and P. rubra L. These species have in common capsule dehiscent fruit type, an exception in Passiflora, whose fruits are commonly berry type. To contribute with new morphological data that may be useful in systematics and phylogenetic interpretations and recognition of genes function related to morphological structures, the objective of this study was to characterize anatomically the fruits and seeds of Passiflora capsularis , P. citrina and P. rubra . For this, flowers were pollinated and after the development of fruits they were collected and processed according to the usual techniques of anatomy.
The data indicate that fruits and seeds of three species have a similar anatomical structure. In uniseriate epicarp, simple trichomes were observed. The mesocarp has several layers of parenchyma cells, in which prismatic crystals and phenolic compounds can accumulate and where are immersed the collateral vascular bundles. The endocarp is formed by a single layer of primary cell walls and has stomata. In the region of dehiscence of the fruit, a vascular bundle of higher caliber and parenchyma were observed. There is no line of dehiscence. The seeds are bitegmic, with outer integument – testa – comprised of two layers of cells and inner integument – tegmen – with three layers. The exotesta has cuboidal cells while mesotesta consists of elongated cells with some intercellular spaces. Exotegmen have palisade cells, constituting a mechanical layer of the seed. In cells of mesotegmen, deposition of phenolic compounds occurs, and endotegmen is compressed. The results confirm the literature data for the family and are of remarkable
562 importance since little is known about the anatomy of the fruits of Passiflora . Finantial support: Fapesp
P0706 – Poster
Fruit micromorphology in the Umbelliferae and its taxonomic significance
Ostroumova, T 1 , Pimenov, MG 1 , Ukrainskaya, UA 1
1 Botanical Garden, Moscow State University, Russia
Fruit morphology and anatomy are very important in the taxonomy of Umbelliferae. In 1971-2008 a number of taxa were investigated with SEM: tribes Caucalideae,
Tordylieae, genera Ammi, Visnaga, Eryngium,
Pimpinella, Ferula, Lichtensteinia . The application of micromorphological characters for taxonomy is difficult owing to the absence of a uniform and standard terminology. One of the problems is the wide range of magnifications possible with SEM, and the similarity of details seen at different magnifications. It is therefore most appropriate to describe the microstructure in three categories: the shape of the cells (the primary sculpture), the fine relief of the cell wall (the secondary sculpture), and the epicuticular secretions (the tertiary sculpture) as proposed by Barthlott & Ehler (1977) and Barthlott
(1981). We studied the micromorphology of the fruit surface of 590 species representing 285 genera and propose the following list of characters. Cell pattern: cells arranged (1) in longitudinal rows, (2) at random.
Cell shape: cell borders may be either clearly visible or indistinct; if cell borders distinct: outline of cells: (1) isodiametric, (2) oblong (length:width 1,5 – 5), (3) long
(l:w>5); anticlinal walls: (1) straight (usual state in
Umbelliferae fruits), (2) curved; relief of cell boundary:
(1) flat, (2) sunken, (3) raised, (4) with narrow groove; curvature of outer periclinal wall: (1) flat, (2) concave,
(3) convex, (4) broadly domed, (5) broadly conical, (6) with a small cone, (7) with a small dome in the cell centre, (8) papillate, papillae being cylindric and much narrower than the epidernal cells, (9) with capitate papillae, (10) columnar – papillae being almost as broad as the epidermal cells. If cell borders indistinct: surface
(1) smooth, (2) longitudinally sulcate (with long regularly spaced grooves), (3) longitudinally rugate, rugae 10–50 µm long, (4) irregularly rugate, (5) foveolate-tuberculate, (6) undulate, (7) with dome-like projections, (8) with conical projections (9) papillose.
Microstructure of the outer cell wall: (1) smooth, (2) sparsely striate, (3) striate with straight striae, (4) striate with undulate striae, (5) striate with tuberculate striae, (6) striato-knotted with striae confluent on conical projections, (7) rugulate (elongated sculpture elements ca. 1 µm in width), (8) sparsely rugulate, (9) 3D striatorugulate, (10) tuberculate with circular elements 1-3
µ m in diameter, (11) rugulate-tuberculate, (12) reticulaterugulate. Epicuticular secretions: (1) wax absent, (2) wax layer rather smooth with small scattered grains, (3) scattered grains and irregular particles, (4) globose particles, (5) complex aggregations of scales. For example, all investigated Anginon and Ostericum species have convex or domed outer cell walls. In Anginon , cuticular striations are often 3
µ m broad; in Ostericum 1
µ m. In Caucalis , Orlaya and Torilis japonica the surface of the secondary ribs is rather smooth, whereas Torilis arvensis , Lisaea and Turgenia have conical projections

and a thick wax layer that hides the cuticular sculpturing; in Agrocharis the conical projection have a clear striatoknotted microsculpture. The hair surface in Seseli is usually rugulate; in Pimpinella it is tuberculate. In this study, many new micromorphological characters were revealed that are taxonomically useful at generic and species level.
P0708 – ePoster
Catalogue of seed plants of the West Indies
Acevedo, P 1 , Strong, M 1
1 US National Herbarium, USA
This catalogue enumerates all taxa of Gymnosperms,
Dicotyledons, and Monocotyledons occurring in the
West Indies Archipelago excluding the islands off the coast of Venezuela (Netherlands Antilles, Venezuelan
Antilles, Tobago, and Trinidad). For each accepted taxon, nomenclature (including synonyms described from the West Indies and their references to publication), distribution in the West Indies (including endemic, native or exotic status), common names, and a numerical listing of literature records are given for each. Type specimen citations are provided for accepted names and synonyms of Cyperaceae, Sapindaceae, and some selected genera in several families including the Apocynaceae ( Plumeria ),
Aquifoliaceae ( Ilex ) and Santalaceae ( Dendropthora ).
Nearly 30,000 names were treated comprising 231 families, 1,945 genera, and 12,275 taxa which includes exotic and commonly cultivated plants. The total number of indigenous taxa was ca. 10,472 of which 71% (7,451 taxa) are endemic to the Archipelago or part of it.
P0712 – ePoster
Plazi /Goldengate as tools for markup and data exchange of taxonomic publications
Sautter, G 1 , Catapano, T 2 , Morris, R 3 , Agosti, D 4
1 Karlsruher Insittute of Technology, Karlsruhe,
Germany; 2 Plazi, Columbia University, New York, USA;
3 Plazi, University of Massachusetts, Boston, USA;
Tehran, Iran
4 Plazi,
Treatments of taxa are the centerpiece of taxonomy.
They are taxon specific, and represent in various forms the entire synthesis of knowledge on a particular species to snippets of it, such as a catalogue entry. They are also a part of what is required to make a new name available in the sense of the Codes that govern nomenclature.
Operationally, it is the place in the literature where scholars and users turn to find the most specific information about a particular taxon. For each taxon, one or more treatments exist in the printed literature and increasingly scattered over the Internet, forming a body of descriptions of the 1.8M non-microbial species currently described. Despite the internet search facilities, most treatments are not available except in printed form, and many of the digitized treatments cannot be found because the search engines are not context sensitive, and thus treatments are listed along with many other pages that include the taxon names. Digitizing and adding markup to legacy publications, including markup in born digital publications, and providing access by online distribution services to the treatments is one solution to this problem. In this contribution we describe the thoughts and decisions that led ultimately to the implementation of such a process developed by Plazi, a
Swiss-based, international not for profit association. The mark-up of legacy publication is dealt with using
GoldenGATE, a highly versatile semi-automatic editor that not only finds and marks required elements such as names, treatments and their substructures like nomenclature, descriptions, materials examined, and keys, but also creates links to external resources by including existing identifiers from dedicated databases such as nomenclators and bibliographies. For legacy literature, the input to GoldenGATE comes from the
Biodiversity Heritage Library, among others. TaxonX, the markup schema used, is a domain-specific Extensible
Markup Language (XML) schema, that does not model the entire publication but the taxa. For prospective publishing Taxpub, an XML Document Type Definition
(DTD) extension of the widely used Journal Publishing
Tag Set by the US National Library of Medicine, has been developed and implemented in at least two journals,
Phytokeys and Zookeys . Once the documents are in
XML, they can be exposed as such and harvested by machines directly. In Plazi, the documents are ingested and stored in a dedicated database allowing not only finding and searching the treatments and data extracted from them, but also distributing them through dedicated services, using the TAPIR exchange protocol for materials examined as implemented with the Global
Biodiversity Facility, or the SPM exchange protocol as implemented with the Encyclopedia of Life. The pros and cons of this approach will be discussed, such as why to chose XML and not the Resource Description
Framework (RDF) language or other ways, as well as the resources needed.
P0714 – ePoster
Molecular phylogenetics of Nepenthaceae based on
1 internal transcribed spacer (ITS) DNA sequences
Alamsyah, F 1 , Ito, M 1
Dept of General System Studies, University of Tokyo,
Japan
Nepenthaceae, a monotypic family of pitcher plants with only one genus, Nepenthes , currently comprises 120 commonly recognised species that occur mainly in
Southeast Asia. Recent works have been conducted on phylogenetic analysis of Nepenthaceae with 71 taxa by using trn K intron as phylogenetic marker. But, comparative analysis between the Nepenthes trn K intron with its translocated copy, revealed an incongruence to tree topology and it was evaluated that the phylogeny of
563

the trn K intron could not reflect true phylogenetic relationships of Nepenthes . PTR1 (peptide transferase 1), a nuclear low copy gene in Nepenthes then introduced as a marker gene, but still it had weaknesses and different position of some taxon in the tree topology from the previous one. So, the question of the origin of the
Nepenthes remained unresolved. ITS region that includes
ITS-1, ITS-2, and the intervening 5.8S gene have been widely used by plant systematists because of its small size, highly conserved flanks and high copy number, and had been used to study the chemical compounds contained in Ancistrocladus tectorius complex, a sister taxon of Nepenthes. Sampling of dried leaves samples from 68 species of Nepenthes in Indonesia included ten new plant species that have never been used for phylogenetic research. ITS region were amplified by
PCR reaction and analyzed using a CEQ8000 automated sequencer (Beckman Coulter), using the same primers as for PCR. Phylogenetic analysis based on Neighbor
Joining method were performed using MEGA 5 by using
Ancistrocladus abbreviatus as outgroup. 3 clades were formed from the ITS datas analysis. The first clades comprises 24 taxa from Sumatra, Java, Borneo,
Malaysian peninsula, and Thailand. There was an assumption based on evidence from molecular data that the colonization of SE Asia started from the Indian subcontinent, marked by the relict N. khasiana which is endemic to northern India. The second and third groups comprise 17 and 6 taxa, respectively, from Borneo,
Sulawesi, New Guinea and the Philippines. There was an assumption that migration proceeded from the
Philippines to Borneo and Sulawesi and also to New
Guinea, where there were probably two land bridges existing from the Philippines to Borneo via Palawan and
Mindanao. The taxa within the clades were congruent with the subdivision of Nepenthes according to Danser.
The taxa are also characterized with their own altitudinal distribution, which divides into lowland and highland plants; life forms, which occur as epiphytes or terrestrial plants; habitat, which comprises mossy forest, shady forest, heath forest, scrub vegetation, and sandstone; pitcher form, which consist of ellipse, cylinder, funnel, and ovate form. These informations could reflect their own character and ecological evolution.
P0715 – ePoster
Xeromorphic anatomical characters and their adaptive value in understory Marantaceae species
Albuquerque, B 1 , Braga, A 2 , Vieira, C 3
1 Centro Universitário Augusto Motta, Rio de Janeiro,
Brazil; 2 Instituto de Pesquisa Jardim Botânico do Rio de
3 Janeiro, Rio de Janeiro, Brazil; Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil
The family Marantaceae has a pantropical distribution, and its species are usually classified as mesophytic and hygrophilous. Nevertheless, the present study has identified a set of xeromorphic characters in understory species. In order to analyze the adaptive value of anatomical characters to the environment, we studied the leaves – sheath, petiole with distal pulvinus, and leaf blade – from 16 species belonging to all 11 Brazilian genera collected in the Atlantic Forest, Amazon and
Cerrado, using standard cytohistological techniques.
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Many of the analyzed species featured heavy epicuticular wax or anthocyanin on the abaxial face of the leaf; epidermal cell walls heavily thick; hypodermis on both faces, usually rich in cellulose mucilage; mesophyll compact; presence of silica druse in the mesophyll; silica and/or wax deposits on the substomatal chamber; silicabodies, such as stegmata, associated with sclerenchyma fibers throughout the leaf axis; and sclerenchymatic hypodermis on the sheath and petiole. It was observed that in tropical forests, light availability and distribution are not constant due to canopy arrangement – in other words, the architecture of foliage and branches, as well as natural clearings. As the analyzed species are adapted to shaded environments and occur in forest edges with direct sunlight during part of the day, it is concluded that the adaptations are a response meant to minimize sunflecks, which are quite common in understory environments. As high radiation levels are commonly associated with increased reproductive activity in understory plant species, it is concluded that Marantaceae
– which are wide-leaved and associated with moist soil environments – are adapted to light and heat exposure as well. This justifies the combination of hygromorphic and xeromorphic anatomical characters in the leaves of the studied species.
P0716 – ePoster
Microsatellites as tools for species delimitation and diagnosis of hybrid origins in the hyper-diverse genus
Boechera
(Brassicaceae)
Allphin, L 1
P 3
, Beck, J
, Al-Shehbaz, I 4
2 , Call, C 1 , Bailey, C D
, Windham, M 2
3 , Alexander,
1 Brigham Young University, USA;
USA; 3
2 Duke University,
New Mexico State University, USA; 4 Missouri
Botanical Garden, USA
Recent dated phylogenies indicate that the genus
Boechera (Brasssicaceae) diverged from its closest relatives less than 2 million years ago. Over the relatively short span of its existence, the genus has experienced a rapid radiation producing 60 + morphologically distinct, sexual diploids. Reproductive isolation has not kept pace with morphological divergence, however, and most of the sexual diploids hybridize whenever they come into contact. These hybridization events have given rise to a diverse array of diploids, triploids and tetraploids, all of which serve to obscure the morphological distinctions among the sexual diploids. Microsatellites (SSRs) are
DNA markers that are becoming increasingly popular for population-level genetic studies and species delimitation in recently diverged lineages . Because of their substantial variability and co-dominant inheritance, they also provide a powerful tool for investigating hybrid origins.
A set of 15 microsatellite loci have proven to be broadly amplifiable and informative, both for delimiting sexual diploid species and diagnosing the genomic constitution of hybrid individuals. We use these microsatellite data, in conjunction with morphology and chromosome counts, to examine the origin(s) and taxonomic status of a variety of species within Boechera . Some taxa and/or populations previously thought to be sexual diploids have proven to be apomictic diploid hybrids. These, in turn, have played a key role in the origin of the many apomictic triploid lineages found in nature . Through the

use of microsatellites markers, we have begun to tease apart the complexity that is Boechera .
P0717 – ePoster
Encouraging online publication of taxonomic and floristic research data using BRAHMS –online
Amaral, M 1 , Bittrich, V 2 , Liddell, A 3 , Filer, D 3
1 University of Campinas, Brazil; 2 Dr Mário de Nucci,
500, 13083-290 Campinas, SP, Brazil; 3 University of
Oxford, UK
The internet provides an ever growing range of vital services for botanists. Data and images from herbaria, specific Floras and taxonomic groups can be published online without delay, at minimal cost, are easily updated and the distribution of these data is maximised. Data online offer far greater flexibility than traditional printed botanical works such as Floras and revisions. Depending on the type of data and the facilities offered by the website, data online can be selectively queried, sorted, mapped and processed into many categories of output including the traditional formats used by current Flora and monograph journal series. A simple and effective approach, as available on http://www.efloras.org (e.g.,
Flora of China ) is the transformation of a paper Flora into pdf format. An important but often missing step in this process is to enable individual researchers who have developed their own research database for a checklist or taxonomic group to easily publish these data online.
Many valuable publications, for example theses on taxonomic groups, remain inaccessible after the work is completed. The need for a more efficient system to promote floristic and taxonomic data from grey literature to web-availability is especially critical in areas where botanical survey and taxonomic work is highly active, often areas with high species diversity and complexity.
As data for such areas are continuously being updated, printed publications have more limited value. The
BRAHMS data management system
(http://dps.plants.ox.ac.uk/bol) is used widely to provide services for botanical research projects. Data from these databases can be selectively published to BRAHMS online (BOL) using BRAHMS WebConnect as described on http://dps.plants.ox.ac.uk/bol/content/documentation/BR
AHMSWebConnect.pdf. Individual botanists with no or minimal website development experience can create and design their own web portal and configure their online system to suit the type of data uploaded. BRAHMS and
BOL are now being further developed to facilitate both the capture of floristic and taxonomic data and their transfer online. The objective is to provide a seamless link between the researcher’s database and the website giving the respective taxonomist full control over website design and data uploading. It is also possible for groups of taxonomists to cooperate with the production of an online Flora or another logical grouping of data. Multiple databases may be connected to a single portal and collectively queried. The new approach is currently being tested using various databases of taxonomic or floristic projects of the Brazilian flora (funded by FAPESP visiting scientist project 2009/50013-6). We suggest that this approach will encourage taxonomists to publish their data online and thus help the scientific community to access, analyze and improve data hitherto unavailable.
P0718 – ePoster
Scape anatomy, fruit dehiscence, and chromosome numbers support polyphyly of
Neomarica
(Iridiaceae) and parallel evolution of the leaf-like scape
Gil, A 1
M 1
, Trad, R 1 , Urdampilleta, J 1 , Bittrich, V 2
1 University of Campinas, Brazil;
Campinas, SP, Brazil
2
, Amaral ,
Dr. Mário de Nucci,
The family Iridaceae Juss. is composed of ca. 66 genera and 2,030 species. The neotropical tribe Trimezieae Rav. is probably monophyletic, but the delimitation of its currently accepted four genera is still uncertain.
Neomarica Sprague is the largest genus of this tribe with ca. 24 species. It is characterized mainly by the broadly flattened scape, morphologically very similar to the leaves, formerly considered a synapomorphy of the genus. Recent studies on fruit morphology, chromosome numbers and phylogeny using DNA sequence data, however, pointed to the possibility that the genus is polyphyletic and its species belong to three not closely related clades. This suggests that the characteristic kind of scape might have evolved three times in the tribe and is a false synapomorphy for the genus. We therefore investigated the anatomy of the scapes to verify if anatomical data would support the idea of parallel evolution of a broadly flattened scape in the groups of
Neomarica . Anatomical sections of the scapes of 12 species of Neomarica belonging to the three clades and of 5 species of Trimezia were made by hand and stained with safranine /astra blue. The sections were immersed in glycerin jelly and on the following day studied and photographed in a microscope. We found two different types of scape anatomy agreeing with the findings of different and not closely groups of Neomarica based on the phylogenetic analysis of DNA sequence data, capsule morphology and chromosome number. One type is characterized by an abrupt constriction of the peripheral parts of the scape into two wings, which may be short or elongated. In most species, this wing formation is correlated with a thick peripheral sclerenchymatic ring in the scape closed in the region where the wings are located. The wings of these scapes are very thin and mainly build of sclerenchyma and vascular bundles. The chromosome number of the plants with this anatomy is
2n=32 and the dehiscence of the fruits occurs only at the apex. In the second type, the peripheral parts of the scape narrow gradually to form elongated wings and the sclerenchymatic ring is open at exactly these regions.
The wings of these scapes are build of parenchyma with scattered vascular bundles. The chromosome number of these plants is 2n=18 and the fruit dehisces, so far as known, completely down to the base. The first type is typical for Neomarica spp. belonging to the group presumably sister to the genus Trimezia and for N. rupestris , which is more closely related to the genus
Pseudotrimezia . The scape of Trimezia spp. is either completely wingless or presents only very short wings.
The sclerenchymatic ring here is always closed, which probably represents the plesiomorphic scape anatomy in the tribe. The second type characterizes species of
565

Neomarica s.s. Our study once more demonstrates that phylogenies obtained from molecular data may provoke interesting character re-evaluations of morphoanatomical characters.
P0719 – ePoster
Genecology, hybrid introgression phylogeography of Icelandic birch
Anamthawat-Jónsson, K 1 and
1 University of Iceland, Reykjavik, Iceland
An overview of researches on Icelandic birch is presented. Two birch species coexist in Iceland, diploid
Arctic dwarf birch Betula nana and tetraploid European tree birch Betula pubescens . Morphologically both species are highly variable and this has rendered taxonomic delineation very difficult. Our studies using botanical, cytogenetic and molecular markers have shown that the variability in this pair of birch species is due to introgressive hybridization via triploid interspecific hybrids.
More than four hundreds birch plants from all major birch woodlands in Iceland have been examined.
Chromosome counts from mitotic metaphases of all individuals under study produce only three groups of birch: diploid, triploid and tetraploid. No aneuploid was found. The three ploidy groups are confirmed by genome size analysis on a subset of samples based on flow cytometry and DNA densitometry.
By looking at the plant morphology, i.e. analysing species-specific botanical characters qualitatively and quantitatively, we can separate the diploid and the tetraploid groups most of the time. The diploid group consisted mostly of dwarf birch B. nana , whereas the tetraploid group predominantly included B. pubescens like plants. The triploid plants, on the other hand, resembled B. nana or B. pubescens but others had intermediate morphology. The study revealed a clear case of bidirectional introgression.
This was supported by analysing haplotype variations in the chloroplast genomes, in comparison with birch samples from outside Iceland. All common haplotypes found in Iceland were shared between the triploid group and the parental species. The statistical analysis of introgression indices and variation components further indicated introgression with a geographical structure. An east–west haplotype distribution within Iceland was observed and this may indicate different population histories or multiple origins of Icelandic birch. But in relation to the samples from Scandinavia, Scotland and
Greenland, Icelandic birch is apparently European in origin.
Icelandic birch is most likely post-glacial in origin, i.e. having colonized Iceland in the early Holocene. The first colonization of Holocene birch undoubtedly occurred in the north and north-eastern valleys and this is thought to be the result of different deglaciation patterns in that area together with early-Holocene warming in northern
Iceland. In order to find out if the introgressive hybridization detected in the present-day birch has been
566 going on for a long time or it is a recent event, we examined pollen samples from an early Holocene peat profile from Hella in Eyjafjördur, mid-northern Iceland.
Birch pollen appeared as early as 10 cal. ka BP, based on known tephra layers. More interestingly, we found evidence of interspecific birch hybrids throughout the profile. The frequency of non-triporate pollen in a period between 9.2 and 8.7 cal. ka BP far exceeded the average level produced by the present-day triploid hybrids.
Climatic and ecological conditions may have favoured hybridisation of birch species during the expansion of birch woodlands in warm periods.
P0720 – ePoster
Holocene hybridisation in Icelandic birch
Anamthawat-Jónsson, K 1
Æ 1 , Hallsdóttir, M 2
, Karlsdóttir, L 1 , Thórsson,
1 Faculty of Life and Environmental Science, University of Iceland, Reykjavik, Iceland; 2 Icelandic Institute of
Natural History, Reykjavik, Iceland
The only tree species creating continuous woodlands in
Iceland during the Holocene is the downy birch Betula pubescens . A related species, the dwarf birch B. nana is also widely distributed in Iceland. Downy birch in northern Scandinavia and Iceland generally has low and scrub-like growth compared to the same species further south. The most probable cause is gene flow via introgressive hybridization between downy birch and dwarf birch, which has been confirmed with experiments and genetical field researches (1, 2). Downy birch is tetraploid with chromosome number 2n = 56, but dwarf birch is diploid with 2n = 28. In natural woodlands in
Iceland, triploid hybrids (2n = 42) deriving from hybridisation between the two species are reletively common. However, little is known about the history of birch hybridization in Iceland, e.g. if the process is continuous or what environmental factors may have promoted the hybridisation and consequent introgression .
Measurements of birch pollen have shown that downy birch pollen grains are on the average bigger than those of dwarf-birch and their pollen pores are also deeper (3).
Pollen grains of triploid hybrids tend to be small like the dwarf birch’s grains, with deep pores like the downy birch’s pollen, resulting in a low diameter/pore (D/P) ratio. Furthermore the triploids produce a lot of abnormal pollen grains (3). We then used these attributes of hybrid pollen for identification of pollen from sediments or peat.
We searched for evidence of birch hybridisation in previously analysed peat from early Holocene or the period from 10.3 to 7.0 cal ka BP, by re-examining a peat profile from Eyjafjördur, North-Iceland. Birch pollen grains were extracted anew, measured and abnormal grains counted. The ratio between downy birch and dwarf birch pollen was calculated for each sample, assuming two normal distributions with different means for grain diameter . The study (4) revealed a low proportion of downy birch pollen in the oldest peat samples and again around 7.8 cal. ka BP, when dwarf birch predominated. The proportion of downy birch pollen peaked approximately at 8.7 and 7.2 cal. ka BP.
Evidence of hybrids was found in several samples, especially simultaneously with the earlier downy birch peak. Pollen with low D/P ratios (hybrid pollen) was

found at different frequencies throughout the profile.
Non-triporate Betula pollen grains were observed in most samples, especially in a period between 9.2 and 8.7 cal. ka BP when its frequency far exceeded the average level produced by the present-day triploid hybrids. Climatic and ecological conditions may have favoured hybridisation of birch species during the expansion of downy birch over dwarf birch colonies in warm periods .
References: (1) Anamthawat-Jónsson K, Tómasson T
1990; (2) Thórsson Æ.Th. et al. 2007; (3) Karlsdóttir L et al. 2008; (4) Karlsdóttir L et al. 2009.
P0721 – ePoster
Not just dates: evolutionary history of the palm genus
Phoenix
Marques, I 1
L 1
, Channing, A 2 , SanmartÃn, I 1 , Anderson, C
1 Real Jardin Botanico, CSIC, Madrid, Spain;
Sciences, Cardiff University, UK
2 Earth
Phoenix L. (Arecaceae: Coryphoidea) is an Old World genus of palms that comprises 13 species. The different species colonise a wide range of habitats, such as edges of mangrove in South East Asia, rainforest and high altitude grasslands in tropical and subtropical Africa, semi-arid scrubland in Macaronesia, northern Africa and the Arabian peninsula, steep cliffs in coastal areas of the
Eastern Mediterranean, and pine forests at 1500 m altitude in northern India and Nepal. The fossil record of
Phoenix is relatively rich, and seeds, leaves, wood and pollen have been recorded in Europe from the Eocene to the Lower Miocene, and in Africa from the Oligocene onwards. Several of the species within Phoenix are commercially and historically important, such as the well-known date palm, P. dactylifera . Yet, despite its economic importance, relationships within the genus remain largely unknown. This is not surprising since, until recently, it has been difficult to develop a phylogenetic classification of the palms due to a slow rate of nucleotide substitution in chloroplast DNA. In this contribution, phylogenetic analyses of seven chloroplast markers ( mat K, ndh F, trn D-T, trn Crpo B, trn Qrps 16, rps 16, trn S-trnfm) from the 13 described species of
Phoenix have been used to answer the following questions: (1) Can these chloroplast markers be used to reconstruct phylogenetic relationships within Phoenix ?
(2) Is the resulting hypothesis of relationships congruent with those of morphological studies? (3) Can a biogeographic scenario of Phoenix be reconstructed with the help of molecular dating and fossil evidence?
P0722 – ePoster
The Missouri Botanical Garden's DNA bank: challenges and opportunities for service to the systematics community
Applequist, W 1 , Kuhlman, A
P 1 , Bussmann, R 1 , Magill, R 1
1 , Solomon, J
1 Missouri Botanical Garden, St Louis, USA
1 , Lowry II,
The Missouri Botanical Garden’s William L. Brown
Center maintains a repository of leaf samples preserved in silica gel that are made available to support molecular studies of plant systematics and evolution . As of October
2010, this collection comprised nearly 11,000 accessioned samples, collected from 55 countries and representing 368 families and 2502 genera . Since its inception in 1998, 1532 samples have been distributed to researchers . The majority of requests are from US-based researchers, although requests have been received from
23 other countries . Fifteen countries are represented by more than 100 samples (the top four being the United
States, Madagascar, New Caledonia, and Bolivia), as are seventeen angiosperm families (by far the most strongly represented being Asteraceae, with 2236 samples and half of the twenty most-sampled genera) . Over 400 bryophyte samples are also available . The collection therefore has great potential to facilitate the efficient and affordable conduct of molecular systematic studies . The distribution of samples reflects the Garden’s international partnerships and collectors’ interests; since such biases are probably inevitable, a network of DNA banks may provide better support to the botanical research community than any single institution could . A successful DNA bank needs to address various questions, including how and in what form to store samples, how to increase collection diversity while maximizing sample quality, and how to control or defray the financial costs of preserving and distributing material . Additionally, the institution must ensure that legal and ethical restrictions on sample use arising from the Convention on Biological
Diversity, national laws, and the terms of collecting and export permits are respected . We describe briefly the
Missouri Botanical Garden’s approach to these challenges and discuss means by which the scientific value and utility of our DNA bank and similar repositories could be further enhanced.
P0727 – ePoster
Reconstruction of ancestral pollen morphologies aids calibrating of the phylogenetic tree of Apiaceae subfamily Apioideae (Magnoliophyta, Apiales)
Banasiak, L 1 , Spalik, K 1
1 Dept of Plant Systematics and Geography, Institute of
Botany, University of Warsaw, Poland
The calibration of phylogenetic trees is crucial for historical biogeography and evolutionary analyses.
However, for many groups of angiosperms including the family Apiaceae, fossil data of their early representatives are scarce or hardly preserved. Moreover, their assignment to extant lineages is often problematic. We attempt to overcome this problem using evolutionary analyses of extant and fossil pollen data for the umbellifer subfamily Apioideae. We have sampled a representative subset of extant members of the subfamily for molecular markers (nrDNA ITS sequence) and pollen morphology. Based on molecular phylogeny, we inferred ancestral morphologies of pollen grains for most major lineages. Then we attempted to assign fossil pollen data to inner nodes of the tree. These assignments were used as calibration points in Bayesian analyses of the nrDNA
ITS data.
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P0728 – ePoster
Characteristics of the woodland vegetation of Tahtalý
Mountains, Turkey
Bani, B 1 , Adýgüzel, N 2
1 Yüzüncü Yýl University, Turkey; 2 Gazi University,
Turkey
Tahtalý Range is situated in the Mediterranean region. It extends c. 150 km from Aladað massif to northeastwards. Beydað, Kýzýlgöl and Soðanlý mountains are included in Tahtalý Range. The area has a semi-arid
Mediterranean climate; the dominant rock of the area is limestone. Coniferous forests of the research area are characterized by Pinus nigra J.F.Arnold. subsp . nigra var. caramanica (Loudon) Rehder, Abies cilicica (Ant. &
Kotschy) Carr. subsp. cilicica and Cedrus libani A.Rich., forming pure or mixed stands; Juniperus excelsa M.Bieb. and J. foetidissima Willd. occur mainly at 1500-1700 m; and Quercus cerris L. var. cerris , Q. libani Oliv., Q. infectoria Oliv. subsp. boissieri (Reut.) O.Schwarz, Q. petraea (Matt.) Liebl. subsp. pinnatiloba (K.Koch)
Menitsky, Q. robur L. subsp. robur and Q. pubescens
Willd. are dominant taxa of deciduous oaks. In the many places coniferous forests destroyed are being replaced by oak and juniper species. In this study, woodland vegetation of the research area is given with pictures.
P0729 – Poster
Fossil flowers, fruits and seeds from the early
Miocene Foulden Maar, New Zealand
Bannister, JM 1 , Conran, JG 2
Mildenhall, DM 4
, Lee, DE 3 , Kaulfuss, U 3 ,
1 Dept of Botany, University of Otago, Dunedin, New
2 Zealand; 2Australian Centre for Evolutionary Biology and Biodiversity, School of Earth & Environmental
3 Dept of Sciences, University of Adelaide, Australia;
Geology, University of Otago, Dunedin, New Zealand;
4 GNS Science, Lower Hutt, New Zealand
Flowers are uncommon as fossils, and examples from the
Southern Hemisphere with in situ pollen are extremely rare. In the laminated diatomite deposit at Foulden Maar, southern South Island, New Zealand a range of flowers
(both uni- and bisexual), inflorescences, buds and clusters of anthers, as well as fruits and seeds are preserved, together with a diverse array of leaves with good cuticular preservation. The occurrence of some flowers with in situ pollen together with a variety of other reproductive parts makes it possible to relate them to extant taxa, as well as confirming the identities of dispersed pollen taxa. Flower families recorded to date include Cunoniaceae, Elaeocarpaceae, Euphorbiaceae,
Lauraceae, Loranthaceae, Monimiaceae ( Hedycarya ),
Onagraceae ( Fuchsia ) and Picrodendraceae as well as probable Alseuosmiaceae and Rutaceae. Flowers range in size from 2 to 20 mm diameter, and most appear to have been insect-pollinated. All were sourced from forest trees and shrubs growing around the perimeter of a 1500 m diameter volcanic maar lake. Some of the flowers are from genera no longer present in New Zealand, whereas others, including Fuchsia and Hedycarya , are still
568 important components of the modern New Zealand flora.
The 23 million year old flowers from Foulden Maar all represent extinct species, as might be expected. Fruits and seeds are also scattered on the bedding planes, generally as isolated fossils, but sometimes as fruit with in situ seeds. As with the flowers, numerous families are represented, including Atherospermataceae ( Laurelia ),
Bignoniaceae, Casuarinaceae ( Gymnostoma ),
Euphorbiaceae, Lauraceae, Leguminosae, Myrtaceae,
Meliaceae, Picrodendraceae, Podocarpaceae
( Podocarpus ), possible Rutaceae and some which are no longer present in New Zealand, such as Menispermaceae.
The diversity and high level of preservation of reproductive materials at Foulden Maar significantly increases our understanding of the Early Miocene flora particularly when combined with associated leaf material for better total evidence approaches to fossil taxa. It also provides insights into the possible biotic associations of the flora for pollination and dispersal in Miocene New
Zealand. Finally, there are implications for co-evolution of the plant families with associated animal groups in the region, including the numerous insects discovered at the same site.
P0730 – Poster
New taxonomic implications in Tillandsioideae
(Bromeliaceae) based on DNA data and morphology
Barfuss, J 1 , Till, W 1
1 Dept of Systematic and Evolutionary Botany, University of Vienna, Austria
In order to establish a natural and stable classification system for Tillandsioideae we conducted phylogenetic analyses using newly generated single-copy nDNA sequences (i.e., PHYC and PRK) and published cpDNA data (i.e., atp Brbc L spacer, mat K, rbc L, partial rbc Lacc D spacer, rps 16 intron, partial trn K intron, trn L intron and trn Ltrn F spacer). Derived phylogenetic units were then characterized using previously used, neglected, and new morphological characters. A new classification for
Tillandsioideae is urgently needed, since modern bromeliad taxonomists rely on a more than 30 years old monograph. Subsequent taxonomic changes were only partly summarized in Floras or recent taxonomic treatments of certain groups. Our results highlight the presence of two new subtribes (Cipuropsidinae,
Vrieseinae), three new genera ( Josemania , Lemeltonia ,
Rothowia ), and three new subgenera ( Racinaea subg.
Pseudophytarrhiza , T.
subg. Viridantha , and T.
subg.
Pseudovriesea ). Beside species from newly erected taxa, several others are also reclassified where initial placement was controversial among bromeliad researchers. Parsimony analysis revealed the following, mostly well-supported general branching pattern:
(
(( Glomeropitcairnia, Catopsis ) ((( Alcantarea, Vriesea )
Werauhia, Cipuropsis–Mezobromelia clade))
( Guzmania , (( Josemania, Racinaea ) ( Rothowia ,
( Lemeltonia, Tillandsia ); Bayesian analysis in contrast revealed a slightly different branching pattern within core
Tillandsieae: ( Guzmania , (( Josemania, Rothowia )
( Racinaea ( Lemeltonia, Tillandsia ))). Relationships of subgenera and informal clades within Tillandsia are mostly unsupported, but most clades themselves receive good statistical support. Although much progress has

been achieved in exploring phylogenetic relationships of
Tillandsioideae by combining both cpDNA and nDNA sequence data, a final and conclusive classification for the whole subfamily cannot be presented here. The most critical groups that remain to be solved using new molecular markers are the Cipuropsis–Mezobromelia alliance and the genus Tillandsia .
P0731 – ePoster
Melastomataceae in Flora Fluminensis, a significant remnant of Atlantic Forest of southeastern Brazil
Baumgratz, JFA 1 , Santos Filho, L 1
1 Instituto de Pesquisas Jardim Botânico do Rio de
Janeiro, Brazil
Tropical forests concentrate the largest proportion of global biodiversity, and the tropical rain forests are the ecosystems with the greatest species richness. Brazil holds about one third of all the remnant rainforests in the world, centered mainly in the Amazonian and along the northeastern and southeastern coastal. The Atlantic
Forest is the second largest forest formation in the
Neotropics and one of the most threatened biomes on the planet. It is also one of the 34 biodiversity hotspots of the world and the fifth richest hotspot of endemic species. In
Brazil, the Atlantic Forest is the second largest ecosystem, with over 14,000 species of vascular plants.
The state of Rio de Janeiro, also known as Fluminensis
State, is the third smallest geopolitical territory in the country (43,696.054 km 2 ) and is entirely located in the
Atlantic Forest biome. In this region there are major mountain ranges such as Serra do Mar and Serra da
Mantiqueira, where forest remnants are well-preserved at higher elevations, besides valleys and hills. The species richness of Melastomataceae in this state has been shown through several taxonomic studies and by the list of
Brazilian plant species recently published. This study is based on herbaria collections and literature data, and botanical material collected during several expeditions.
The family Melastomataceae is one of the most diverse in Brazilian flora, with 68 genera and more than 1,300 species. In the Atlantic Forest it is also very expressive, with 40 genera and 581 species, of which 392 endemic to this ecosystem. The vegetation types of the state include submontane, montane and lowland forests, high altitude grasslands, and restingas (savannas). The diversity of
Melastomataceae in the Fluminensis flora is very significant, with 29 genera and 334 species, representing ca. 43% and 26%, respectively, of total genera and species in the country. The degree of endemism is also very significant, since two genera ( Bisglaziovia and
Itatiaia ) and 97 species are endemic, representing 17% of the total species of Melastomataceae occurring in the ecosystem. The largest genera in species are Leandra,
Miconia and Tibouchina . Most species are trees, treelets, shrubs and subshrubs, sometimes herbs, rare epiphytes.
In the woods there are shrubs and arboreal specimens, mainly of the genera Clidemia, Huberia, Leandra,
Meriania, Miconia, Ossaea and Tibouchina . On the other hand, herbaceous and subshrubs species of Aciotis,
Acisanthera, Bertolonia and Salpinga , usually found in moist and shaded areas, are less frequent. Chaetostoma,
Lavoisiera, Microlicia and Trembleya are more characteristic of altitudes and usually in grasslands areas.
In the restingas occur 11 genera and 22 species, of which four endemic of the fluminensis flora. In the state of Rio de Janeiro there is one of the oldest Conservation Units of the country, as the National Parks of Itatiaia and Serra dos Órgãos, and several species can be identified as critically endangered or another level of threat.
P0732 – ePoster
Melastomataceae in conservation units from theAtlantic Forest, southeastern Brazil: diversity and conservation
Baumgratz, JFA 1
MLD 2
, Santos Filho, LAF 1
, De-Polli, BM 1
, Rei Souza,
1 Instituto de Pesquisas Jardim Botânico do Rio de
Janeiro, Brazil; 2 Universidade Federal de Santa
Catarina, Brazil
Floristic inventories and knowledge of biological diversity are considered priorities in tropical regions, mainly in Conservation Units (CUs) from Brazil, where well-preserved vegetation and families numerically significant, with expressive morphological and taxonomic diversity as Melastomataceae, are found.
Promoting the knowledge of biodiversity in these environments becomes an important strategy for the preservation of ecosystems. The Atlantic Forest is one of the global hotspots of biodiversity, mainly due to the high degree of diversity, endemism, and threat. The state of Rio de Janeiro is located in the Atlantic Forest biome, where some CUs were created. Melastomataceae is very characteristic in this biome and identified as a family of expressive richness of species that occur in montane moist forests, high altitude grasslands, lowland moist forests, restingas (savannas) and coastal islands. This family also shows a wide floral and fruit morphology associated with numerous shapes and colors, which are important attributes of plants that interact in the local ecosystem. Taxonomic studies of Melastomataceae in
CUs have shown a great diversity of taxa in these areas legally protected, with endemic genera and species of these units, as well as to the Atlantic Forest and Rio de
Janeiro State. This study is based on literature data, herbarium collections, and botanical material and information obtained during several scientific expeditions. The National Parks of Serra dos Órgãos and
Itatiaia, the oldest in the country, and the Ecological
Reserve of Macaé de Cima are examples of areas with this species richness and are located in two major mountain ranges of the state, the Serra do Mar and Serra da Mantiqueira, between 100 and 2800 m. In Itatiaia park are found 17 genera and 111 species, of which 68 are endemic to the Atlantic Forest, four endemic to the state of Rio de Janeiro and only the monotipic genus Itatiaia endemic to the area. In Serra dos Órgãos Park are recorded 21 genera and 155 species, of which the monotypic genus Bisglaziovia and five species are endemic to the area, 148 species endemic of the Atlantic
Forest, and 32 species to the Rio de Janeiro State. In
Macaé de Cima Reserve are found 13 genera and 110 species, of which four are endemic to the area, 92 endemic to the Atlantic Forest, and 42 endemic to the
Rio de Janeiro State. For the three areas, the largest genera in species are the same, Leandra , Miconia and
Tibouchina . The incorporation of new areas in the CUs
569

has increased the representation of the family, as well as new records of species, including endangered and endemic species to the state of Rio de Janeiro. On the other hand, some species have not been collected in these areas for over 100 years and, probably, they occur in areas difficult to access or have disappeared due to changes in the original vegetation by human activities.
P0733 – Poster
Phylogeny and taxonomic treatment for
Salvia
sect.
Polystachyae/Iodanthae/Purpureae morphology and pollen
(Lamiaceae):
Bedolla-García, B 1 , Lara-Cabrera, S 1
1 Universidad Michoacana de San Nicolás de Hidalgo,
Mexico
Salvia subgenus Calosphace is considered a natural group with ca. 500 spp endemic to the Americas. The last comprehensive treatment for the subgenus dates to 1939, where Epling classified the species into sections. The sectional classification has been considered artificial by several researchers. In a preliminary phylogeny for
Calosphace , Salvia polystachya (Sect. Polystachyae ), S. iodantha, S. arbuuscula and S. townsendii (Sect.
Iodanthae ), and S. purpureae (Sect. Purpureae ) were monophyletic. In the present study the monophyly of the group is evaluated including 22 species of the three sections. 74 specimens (BIGU, BM, IBUG, EBUM,
ENCB, FCME, IEB, K, MEXU, MO) including type specimens for several species were morphologically evaluated. Twenty nine characters based on vegetative and reproductive structures were evaluated as well as the shape of the pollen grain. Maximum parsimony consensus tree of 10,000 equally parsimonious trees supports the monophyly of the group with 73% of boostrap and 72% of jackniffe support, none the less the dissolution of the sections proposed are not supported.
The synapomorphies that define the group of three sections are the length of the lower corolla lip with respect to the upper one, the compact appearance of the inflorescence and the length of the corolla tube with respect to the calyx. Despite these characters not being exclusive for this group, they do help differentiate the group in conjunction with other characters such as: ovate to ovate-lanceolate leaves but never lanceolate, rounded to attenuate leaf base, deciduous to late deciduous bracts; plus all the species but two, S. polystachya and S. purpurea , are endemic to Mexico. Upon re-evaluation of the sections, previously known sections Iodanthae and
Mexican members of Purpureae are fused into one section Iodanthae , characterized by having ovate to ovate-lanceolate leaves, 10 to 37 (6) flowers per verticilaster, calyx teeth truncated to ovate-acuminate, upper calyx teeth with three veins, corollas from 15 to 32 mm of length, in pinkish to purplish tonalities, exserted style and inserted to exserted stamens. Newly described species S. purepecha and S. mcvaughii are sister to the
Iodanthae clade and classified in a new section
Purepechae ; characterized for having an articulated petiole base, bracts from 1.5 to 8 mm, bracts lanceolate to widely ovate, subpersistent to early deciduous; calyx from 2.4 to 6 mm, tomentose to puberulent, with calyx teeth caudated and the lower teeth proximal, upper calyx teeth with three veins, corolla from 8 to11mm with
570 corolla lips subequal in blue tonalities. Finally Sect.
Polystachyae will be maintained defined by having ovate to ovate-lanceolate leaves, rounded to slightly heart-shaped base, but never entirely heart-shaped, 5 to 8
(28) flowers per verticillaster, calyx teeth ovate-apiculate to acute-apiculate, corolla blue to whitish with lips subequal and inserted stamens. Only subsection
Glabratae is supported. Morphological and pollen evidence proved to be an aid in defining sections and to provide a better classification. The taxonomic treatment recognizes 17 species and eleven species are synonimized.
P0737 – ePoster
Drivers of speciation in large rainforest Myrtaceae genera
Bernardini, B 1 , Forest, F 1 , Hawkins, J 2
1 Royal Botanic Gardens Kew, UK; 2
Reading, UK
, Lucas, E 1
Reading University,
Myrtaceae is an unusual angiosperm family in containing four genera comprising more than 500 species. Three of these, Eugenia (c. 1020 species), Syzygium (c. 1040 species), and Myrcia s.l. (c. 750 species) are predominantly tropical rainforest trees with fleshy fruits.
In each of these genera, rapid shifts in diversification rates have been demonstrated, likely due to evolution of key morphological innovations, abiotic factors, niche partitioning and founder effects in rapidly changing landscapes and climates. However, no hypotheses regarding specific factors driving divergence of large rainforest taxa have been rigorously tested and there is little understanding of the extent to which these factors have shaped tropical rainforest biomes as we know them today . The largest subfamily of Myrtaceae, Myrtoideae, contains all but 2 of the family’s genera. Myrtoideae is composed of 15 tribes between which, phylogenetic relationships have been partially clarified (Biffin et al.
2010). Within Myrtoideae a well supported clade known as ‘BKMMST’ contains tribes Backausieae, Kanieae,
Metrosidereae, Myrteae (including Eugenia ), Syzygieae
(including Syzygium ) and Tristanieae. Phylogenetic relationships of the tribes within BKMMST are not yet fully resolved . Phylogenetic analysis of DNA sequence data (ITS, mat K, ndhF, rpl16) from a taxonomic sample representing tribal species richness and geographical distribution is presented for the first time for the
BKMMST clade. Divergence times are estimated using a
Bayesian relaxed molecular clock (BEAST). Presence and position on the phylogenetic tree of significant diversification rate shifts are assessed using a topological method (SymmeTREE) and the MEDUSA method, which integrates phylogenetic information about the timing of splits and taxonomic richness. Reliably positioned radiation event(s) with clearly identified diversification rate shifts among the branches of the
BKMMST clade can be correlated with single or multiple, biotic and abiotic characters to determine factors driving hyper-speciation. This process is also discussed.

P0738 – ePoster
The enigmatic genus
Aspidoglossum
E. Mey.
(Apocynaceae: Asclepiadoideae–Asclepiadeae) an overview of this complex African endemic
Bester, S P 1 , Nicholas, A 2
1 South African Biodiversity InstituteSouth African
National Biodiversity Institute, South Africa; 2 School of
Biology & Conservation Science, University of KwaZulu-
Natal, South Africa
The genus Aspidoglossum , an endemic to Africa, is widely distributed throughout southern and tropical
Africa. Found mainly along the eastern side of the subcontinent, it has its highest diversity in southern
Africa. The genus was first described by E. Meyer in
1838 and at the time consisted of only three species: A. biflorum, A. heterophyllum and A. fasciculare . These were subsequently all moved to Schizoglossum by R.
Schlechter (1894–1895). In 1984, Kupicha re-instated the genus and enlarged it to 34 species by adding six new species to the original three of Meyer and twenty-five species which have basionyms in Schizoglossum . She also divided the genus into four sections namely sect.
Aspidoglossum , sect. Latibrachium , sect. Verticillus and sect. Virga . Since Kupicha's revision two new species have been added to the genus. The genus (sensu
Kupicha) is thought to be closely related to
Schizoglossum but distinguished from it by the sessile, fasciculate inflorescences, and sausage-shaped pollinia which are sub-apically attached to the corpusculum and equipped with a well-developed germination zone. The species are all perennial herbs with milky latex in all parts. Flowering shoots are produced annually which die back or are burnt to ground level. The plants survive as an underground spindle-shaped tuber. The first author is currently studying Schizoglossum sens. lat. and allied groups which include Aspidoglossum . In this overview the genus is grouped into the current four sections. The distribution of the species, flower morphology, diagnostic characters and a synopsis of species are presented. A number of new species are in the process of being described and some synonyms will be re-instated.
P0740 – ePoster
The Pl@ntnet project: plant computational identification and collaborative information system
Barthélémy, D françois, M 4
1 , Boujemaa, N 2
, Joly, A 2
,
, Mouysset, E 3
Mathieu, D 3 , Jean-
1 CIRAD - UMR AMAP, Montpellier, France;
Imedia Project, Rocquencourt, France;
Montpellier, France;
France;
4
3
2 INRIA,
Tela Botanica,
5
IRD – UMR AMAP, Montpellier,
INRA - UMR AMAP, Montpellier, France
Among the world’s living species, there are approximately 300,000 species of plants. Accurate knowledge of their identity, geographic distribution and use, underpins the success of the world sustainable agriculture with advanced research in agronomy, and biodiversity conservation. Unfortunately, the quality and quantity of plant information is poorest in the regions that could use it the most: Mediterranean and tropical countries. It’s for this reason that plant identification, sharing of and access to plant information play a crucial but little understood role in modern society. In this context, Agropolis Fondation (http://www.agropolisfondation.fr/) is supporting its first Flagship Programme called Pl@ntNet. This programme involves a large international partnership jointly led by the AMAP Joint
Research Unit (http://umramap.cirad.fr), the IMEDIA
Research Team (http://www-rocq.inria.fr/imedia/), and the non-governmental organization Tela Botanica
(http://www.tela-botanica.org/). The Pl@ntNet project aims to set up a web-oriented scientific, informative and educational software platform dedicated to plant identification and to the collaborative gathering, share and use of large, multi-disciplinary datasets on plants.
Free, open-source, easy-access software programs, based on previously developed prototypes will function on-line on internet, or directly in the field, and will be adapted to the various working conditions of potential users (natural resources managers, foresters, agronomists, teachers, scientists or citizens). Plant identification tools will be based on the use of morphological, ecological or geographical characters, as well as on visual image recognition system . As an initiative of major scientific and strategic importance, Pl@ntNet is expected to bring scientific breakthrough and demonstrate the degree and extent by which the know-how, expertise and competencies of various research units and their partners are mobilized and put into use in order to contribute in addressing major challenges of our time.
P0743 – ePoster
Genome-level evidence of adaptive variation among natural eucalypt populations: implications for evolution and conservation
Bradbury, D 1, 2 , Krauss, S 3 , Smithson, A 3 , Veneklaas, E 3
2
1 University of Western Australia, Perth, Australia;
Kings Park and Botanic Garden, West Perth, Australia;
3 Centre of Excellence for Climate Change Woodland and
Forest Health, Australia
Understanding the genetic basis of local adaptation, together with identifying ecologically relevant genomic regions, is a major goal of evolutionary, ecological and conservation genetics. By understanding adaptation at the genomic level we can answer questions about i) species evolution, ii) conservation management of contemporary gene-pools, and of ecological and evolutionary processes, iii) predicting species’ responses to climate change, and iv) the nature of environmental drivers of adaptation.
Simple, rapid methods for identifying adaptation at the genome level are rare, especially for natural populations of non-model species, but the increasing amount of whole-genome information for Eucalyptus has allowed us to investigate new methods for detecting evidence of natural selection. Our aims in this study are to: i) develop gene-associated microsatellite markers that occur within
Eucalyptus expressed sequence tags (ESTs); ii) predict the phenotypic function of those regions by sequence homology with known genes; iii) compare genetic structure and diversity measured with neutral microsatellites versus gene-linked EST-microsatellites; iv) perform statistical tests for natural selection; and v) establish the first reciprocal transplant field trial of our
571

study species to assess variation in quantitative traits.
The study species is Eucalyptus gomphocephala
( Symphyomyrtus ), a forest and woodland tree of ecological value and conservation concern native to the coastal plain of southern Western Australia . We designed and optimised PCR primers for 11 microsatellite loci within Eucalyptus ESTs that are homologous with genes involved in flowering time (CONSTANS), pathogen defence (
β
-1,3-glucanase), heat shock (HSF), photosynthesis (chlorophyll A/B binding protein), desiccation protection (Lea14) and others. Through
Bayesian Fst outlier tests and other statistical methods, we will test for signatures of natural selection at these loci, and based on the sequence homology, will predict the environmental factors that may be involved in the adaptive process. The 1.5 year-old reciprocal transplant trial has revealed phenotypic plasticity among 6 provenances growing at 4 sites located across the species’ range, where limited evidence of local adaptation has been observed so far for seed emergence, survival and growth. Dry-warm provenances, however, exhibit better visual condition after summer than coolwet provenances, regardless of planting location. These results may contribute to the alteration of current guidelines for conservation management and seed collection in an ecological restoration context, and will contribute potentially powerful information towards understanding the nature of eucalypt evolution.
P0744 – ePoster
Molecular systematics of the
Eutrema edwardsii complex (Brassicaceae)
Göbbeler, K 1 , Bruederle, L 1
1 University of Colorado, Denver, USA
Following a major taxonomic revision, genus Eutrema
(Brassicaceae) is currently recognized as comprising 26 species worldwide, most of which occur in Asia . In
North America, the genus is represented by Eutrema salsugineum (Pallas) Al-Shehbaz & Warwick
[ Thellungiella salsuginea (Pall.) O.E. Schulz] and
Eutrema edwardsii R.Br
. Eutrema penlandii Rollins, which is endemic to the Mosquito Range of the Southern
Rocky Mountains (Colorado, USA) and federally listed as threatened, was subsumed into E. edwardsii as part of this treatment . However, population genetic analyses of allozyme data and multivariate analyses of morphological data do not support the synonymization of
E. edwardsii and E. penlandii. We provide the first comprehensive molecular systematic study addressing this problem . Sequences were obtained for four marker genes (nuclear ITS and ETS and chloroplast ndh Jtrn L and trn TL) . Data were collected from a broad sample of over 60 E. edwardsii sites distributed across this species’ range in the Arctic, from Siberia, Alaska, northern
Canada, Greenland, and Scandinavia, as well as most known locations of E. penlandii. Haplotype network analyses, Bayesian Inference, Maximum Likelihood,
Maximum Parsimony, and Neighbornet analyses were used to assess systematic relationships of these populations . Preliminary results indicate well-supported monophyly of E. penlandii clustering within a paraphyletic E. edwardsii. All specimens of E. penlandii share common mutations, insertions or deletions for the
572 investigated marker genes, which can therefore be regarded as molecular synapomorphies for this taxon .
Haplotype network results differ slightly for the diverse gene markers . A common haplotype was found for E. penlandii , whereas E. edwardsii is represented by 6 to 8 different haplotypes . Analyses of the overall more conserved chloroplast marker genes recovers E. penlandii separated from all (in ndh Jtrn L) or most (in trn TL) haplotypes of E. edwardsii species . However, haplotypes of both species are more similar when considering the overall more variable nuclear marker genes . These results further support the hypothesis that
E. edwardsii and E. penlandii are indeed closely related, but distinct taxa . Implications for their systematic status are discussed.
P0746 – ePoster
New insights into the generic circumscription of
Helieae (Gentianaceae)
Calió, M F 1 , Lepis, K B 2 , Pirani, J R 1 , Struwe, L 2
1 Universidade de São Paulo, Brazil;
University of New Jersey, USA
2 Rutgers, State
Helieae is a monophyletic tribe within Gentianaceae. It comprises 23 genera and over 200 species, all endemic to the neotropics. An initial phylogeny of Helieae shed light onto the relationships between genera and indicated that traditional generic limits did not correspond to monophyletic groups. In order to obtain a more thorough understanding of generic relationships within the group, we reconstructed a more comprehensive phylogeny of
Helieae, by enhancing sampling within the so-called
' Symbolanthus -clade'. This phylogeny used DNA sequences from two nuclear regions (ITS and 5S-NTS) and morphological data, which were analyzed separately and in combination using parsimony and Bayesian approaches. A total of 86 individuals representing 17 genera and 51 species of Helieae were sampled – while
47 individuals included sequences from both molecular markers, eight included only morphological characters.
Morphological characters were further mapped onto the total evidence topology to identify potential synapomorphies of clades and patterns of homoplasy in the morphological dataset. Both molecular datasets yielded strongly supported clades, while the morphological dataset resulted in a poorly supported topology overall. The total evidence topology indicated that Prepusa is sister to the remainder of Helieae.
Furthermore, the ' Macrocarpaea -clade' and the
' Symbolanthus -clade' were also recovered; however, on this study the ' Symbolanthus -clade' received higher nodal support on the total evidence topology as compared to the previous phylogenetic study on the tribe. Within the
' Symbolanthus -clade', our results confirm that
Chelonanthus and Irlbachia are not monophyletic, and also contest the monophyly of Calolisianthus as currently circumscribed. Specifically, two species of
Calolisianthus group with the type species of
Chelonanthus , while the other Calolisianthus species are more closely related to Tetrapollinia and Symbolanthus .
Moreover, the white-green-flowered Chelonanthus species are undoubtedly related to Helia . The addition of new characters and taxa led to higher confidence in the relative position of some clades, as well as provided

further support for a new generic circumscription of
Calolisianthus, Chelonanthus, Helia , and Symbolanthus .
Even though several morphological characters traditionally used in the taxonomy of the group were shown to be homoplasious, most clades could be diagnosed by a combination of plesiomorphic, homoplasious, and autapomorphic character states.
P0747 – ePoster
Phylogeny and biogeography of
Anemopaegma
Mart. ex Meisn. (Bignonieae, Bignoniaceae)
Calió, M F 1 , Winkworth, R C, Lohmann, L G 1
1 Universidade de São Paulo, Brazil
Anemopaegma Mart. ex Meisn. is the third largest genus of the tribe Bignonieae (Bignoniaceae). The genus includes 45 species of shrubs and lianas that occurr in both humid forests and dry areas from Mexico to southern Brazil and Argentina. Most species of
Anemopaegma are restricted to a single habitat or vegetation type, and the distributions of several species closely match the patchy distribution of nutrient-poor soils in the Amazon. In order to understand the biogeographic history and patterns of evolution within
Anemopaegma we are currently reconstructing phylogenetic patterns based on two molecular markers, the chloroplast ndhF and the nuclear pepC. Initial results indicate that A. puberulum is sister do the remainder of the genus, which consists of four main clades – two strictly Amazonian (the ‘oligoneuron-setilobum’ and
‘robustum-ionanthum’ clades), one restricted to dry areas
(‘arvensis-album-parkeri’ clade) and one including species that are broadly distributed in humid Neotropical forests (the ‘chrysoleucum-flavum’ clade). Mapping habitat preferences onto the molecular phylogeny suggests that ancestrally Anemopaegma occurred in humid forests and that there was a single transition to dry habitats. This suggests that niche conservatism has played an important role in the evolution of the genus.
P0748 – ePoster
1
Fossil Leguminosae from Mexico: an update on the regional paleodiversity of the family
Calvillo-Canadell, L 1
M 1 , Rico-Arce, L 2
, Cevallos-Ferriz, S RS 1
, Hernández Damián, A L 3
, Sousa,
Departamento de Paleontología, Instituto de Geología,
Universidad Nacional Autónoma de México, Ciudad
Universitaria, Coyoacan, México; 2 HLAA, Royal Botanic
Gardens Kew, UK; 3 Departamento de Biología, Facultad de Ciencias, Universidad Nacional Autónoma de México,
Coyoacan, México DF
Presence of Leguminosae during the Cenozoic in México has been documented since ca. 2 decades ago. They have been identified based on leaves, woods, fruits, flowers and pollen, and most have been preserved as impression/compression, permineralized and amber included fossils. These fossil represent plants closely related to extant ones. For example, from the Middle
Eocene Senna, Chamaecrysta and Inga are represented by compressed fruits and leaves from La Carroza
Formation, La Popa, Nuevo León; Inga, Acacia,
Prosopis, Pithecellobium, Sophora, Mimosa, Lysiloma,
Cladrastis, Apuleia, Piptadenia, Stryphnodendron and
Robinia are represented as compressed leaflets and/or fruits from Oligocene of the Coatzingo Formation,
Puebla. Neogene records from the Miocene El Cien
Formation, Baja California Sur, based on wood samples resemble that of plants of Mimosa, Andira and
Copaifera , and that of Prioria and Dalbergia from the
Tehuacán Formation. Leaflets and woods from unnamed
Miocene sediments near Tlaxcala, Tlaxcala, include
Caesalpinia and Lonchocarpus , respectively. Finally,
Miocene records of Chiapas include those included in amber, like Hymenaea, Albizzia/Leucochloron , and
Mimosa , and compression/impression fossil recently collected in the Ixtapa Formation, have been related to members of Mimosoideae. Analysis of this record, along with geological evolution of Mexico, suggest that diversification of Leguminosae is closely related to physiographic changes that were important in central and southern México during the last 23 my. In contrast, the
Paleogene records seem to be related to the geographic expansion of some taxa to the south, and perhaps, physiographic evolution in this area during these times also influenced diversification and dispersion of the groups. Leguminosae diversification at two distinct times in Mexico closely correlates with geologic evolution of these areas (north vs south), a situation that suggest the strong and direct influence of continental evolution in the relief as the plant were colonizing the new exposed areas that were offering plants not only new sceneries, but sceneries with different condition, that promoted biological response and speciation, generating the biodiversity that is so characteristic of México today.
P0749 – ePoster
1
On important roles of historical specimens at the herbaria for bryological researches
Cao, T 1 , Guo, S 1 , Yu, J 1
College of Life and Environmental Sciences, Shanghai
Normal University, China
Since 1978 when I started my research on bryophytes, I have had opportunities to visit the important herbaria for bryological researches in the world and to undertake researches at some herbaria, including BM, E, PC, H, S,
MHA, LE, KRA, BP (Europe); NY,MO, FH, CANM,
ALTA, MICH (North America); NICH, HIRO, TNS,
LWG, SING (Asia) etc. I was impressed by the rich and valuable collections of bryophyte specimens and literature deposited at the different herbaria. Because of rapid development of new and modern technology, especially molecular methods recently, people pay more and more attention to new technology and molecular analysis. The historical specimens at the herbaria seem not as important for research, especially systematic research on bryophytes, as before. Some people wonder what roles the historical specimens play in the research on bryophytes . Based on my research practice and experience for more than 30 years in bryology, I would like to give my opinions about the important roles of historical specimens at the herbaria for bryological researches herewith. I believe that the historical specimens of bryophytes at the different herbaria are the
573

common treasure of the people of the world. The historical specimens have played, are playing, and will play important roles in bryological studies in different aspects: (1) Taxonomy and classification of bryophytes is the base for any bryological researches. The historical specimens provided solid base for taxonomic and systematic studies of bryophytes. For fully and well understand any group of bryophytes (family, genus, species etc.), you have to study the historical specimens concerned, especially type specimens. My research on worldwide revision of genus Ptychomitrium and
Grimmiaceae in China are the samples to show it. (2)
The historical specimens provide rich and unique material for molecular studies in bryological systematics.
Besides the fresh material, the historical specimens collected during certain periods of time also can be used in molecular experiments for systematic research. There are a lot of good samples about. (3) Analysis of the concentration of chemical elements in the historical specimens collected from different time at different localities can indicate the quality and changes of the environments in certain regions. For example, our results of analysis of the concentrations of heavy metals, Cu,
Pb, Cr, Cd and Zn in the plants of moss Haplocladium collected from different localities in Shanghai City during the past 40 years showed well the spatial and temporal changes of heavy metal concentrations in the past 40 years in the city of Shanghai. (4) The historical specimens also can provide useful information for conservation of biodiversity of bryophytes. Based on checking the historical specimens and comparison with recent collections, the extinct and endangered species of bryophytes in certain regions have been evaluated. In conclusion, we should stress and bring into play important roles of the historical specimens in further research.
P0750 – ePoster
Saprobic fungi diversity of differents environments in
‘Parque Estatal Agua Blanca’ Macuspana, Tabasco,
Mexico
Cappello Garcia, S 1 , Rosique-Gil, E 1 , Cifuentes-Blanco,
1
J 2 , Gama, L 1
DACBiol, UJAT, Mexico; 2 Facultad de Ciencias,
UNAM, Mexico
The study of fungi in Mexico has been conducted mainly in temperate regions, and studies in tropical regions are scarce and focus mainly on macroscopic fungi. This is the first study on the macro- and microscopic fungi saprobes of different environments in a Protected Natural
Area (an area slightly impacted) in a tropical region of
Mexico. Were collected, described and preserved specimens of macroscopic fungi following the regular collection techniques. Litter samples were collected, including several species of palm, and samples were placed in moist chambers and preparations were made semi-permanent structures of fungi for identification.
Soil samples were collected and processed by two techniques: dilution and soil washing, using malt extract agar to isolate fungi in the soil particles. Wooden blocks were placed as bait to pine mushroom extract freshwater, wood blocks were placed in moist chambers and semipermanent preparations were made of the structures of
574 fungi for identification. Some of the fungi that have been identified so far are: Dacryopinaxs elegans, Cookeina tricoloma, Deflexula fascicularis, Oudemanciela afin canary, Phillipsia domingensis, Poliporus arcularius,
Earliella scabrosa, Exserticlava triseptata, Beltrania rómbica, Subulispora longirostrata, Sporidesmium anglicum, Wiesneriomyces indicus, Zigosporium echinosporum, Zigosporium oscheoides, Dictyosporium digitatum, Menisporopsis novae-zelandiae .
P0751 – ePoster
Systematics, evolution, and biogeography of the western North American cleomoids (Cleomaceae)
Cardinal-McTeague, WM 1
Hall, JC 1
, Sytsma, KJ 2 , Hahn, WJ 3 ,
1 University of Alberta, Canada;
Wisconsin–Madison, USA; 3
2 University of
Georgetown University, USA
Cleome sect. Peritoma, Cleomella, Oxystylis , and
Wislizenia are a fascinating group of Cleomaceae with unresolved but important systematic, biogeographical, and morphological questions . Although these taxa form a strongly supported monophyletic lineage called the
Western North American Cleomoids, previous phylogenetic analysis suggested that Cleomella is paraphyletic within the clade. Moreover, competing evolutionary hypotheses have been proposed for the relationship of Oxystylis and Wislizenia , the age of their putative split, and the degree of genetic divergence .
These two monotypic genera are endemic to desert regions of N. America and exhibit highly unusual fruits relative to the rest of the family . Whereas most members of Cleomaceae have many-seeded dehiscent capsules, fruits of Oxystylis and Wislizenia are indehiscent with few seeds . Lastly, most species of the Western North
American Cleomoids are endemic to one or more of the western North American deserts, presumably diverging in situ . Combining timing of phylogenetic relationships with geological history of the region should provide insight into the patterns and process of divergence within the Western North American Cleomoids . Here we present a comprehensive phylogenetic analysis of the clade with exhaustive taxon sampling . Combined nuclear and chloroplast sequence data confirm the paraphyly of
Cleomella. In addition, the Western North American
Cleomoids appear to be a young clade that has undergone a single reduction in the fruits of the Oxystylis plus
Wislizenia lineage.
P0752 – ePoster
Lomelosia crenata ssp. pseudisetensis
(Dipsacaceae) at
1 the southern limit of its distribution (S Italy)
Caruso, G 1 , Uzunov, D 1
Dept of Environmental and Crop Science, Marche
Polytechnic University, Italy
Lomelosia crenata (Cirillo) Greuter & Burdet
(Dipsacaceae) is a southern Italian endemic. Three subspecies belong to this taxon: L. crenata ssp. crenata occurring in Southern Italy and Sicily on limestone, as well as L. c.
ssp. dallaportae (Boiss.) Greuter & Burdet

occurring in Apulia, and L. c. ssp. pseudisetensis
(Lacaita) Greuter & Burdet occurring in central and southern Italy mainly on sedimentary substrata where clasts are cemented by limestone. The southernmost area of the Italian peninsula, located inside the administrative territory of the Calabria region, encompasses mountain ranges located as follow in north–south direction:
Pollino, Orsomarso, Catena Costiera, Sila, Serre,
Aspromonte. Pollino and Orsomarso, as well as the rest of northward Apennine Range, are mainly calcareous, while the rest (Catena Costiera, Sila, Serre and
Aspromonte) are mainly made up of non-calcareous rocks. In the non-calcareous sector of central and southern Calabria some calcareous or conglomerate tectonic windows occur. The survey result suggests that the investigated taxon frequently and continuously occurs on Pollino and Orsomarso limestone, commonly between
100 and 1500 m a.s.l., while on the southern part of the investigated area L. crenata ssp. pseudisetensis is hosted inside some of the tectonic calcareous or conglomerate windows occurring there. Disjunct populations of the studied subspecies have been found southwards of its main range. For this populations chorological and taxonomical studies are in progress and the preliminary data are presented.
P0753 – ePoster
Retama raetam
ssp. gussonei
(Fabaceae): population survey in Calabria (S Italy)
Caruso, G 1 , Uzunov, D 1 , Gangale, C 2
1 Dept of Environmental and Crop Science, Marche
Polytechnic University, Italy; 2 Museum of Natural
History and Botanical Garden, University of Calabria,
Italy
Calabria is the southernmost region of the Italian peninsula. It encompasses a few mountain chains as part of Apennine Range and it is surrounded by a 740 km coast line, most of which sandy, by the Ionian Sea and the Thyrrenian Sea. Climate on Calabrian coasts is mainly themomediterranean. Retama raetam (Forssk.)
Webb. subsp. gussonei (Webb) Greuter (Fabaceae) is a
Sicilian–Calabrian endemic occurring on sandy substrata by the sea or, less frequently, inland. A survey of
Calabrian populations of this species has been carried out. Two R. gussonei subpopulations were already known for the Calabrian territory, the first occurring on the Ionian coast inside Croton e province (Cirò municipality), the second on the Thyrrenian coast occurring partially inside Vibo Valentia province
(Nicotera municipality) and partially inside Reggio
Calabria province (S. Ferdinando municipality). The morphological structure of different aged individuals has been studied in order to establish an age/morphology connection. Individuals have been categorized in different classes on the base of the canopy diameter: class 1 0,5–1 m, class 2 1–2 m, class 3 2–3 m, class 4 3–
6 m. On the basis of this morphological classification the whole population has been censussed. The total number of individuals has been 1523, 82.5% of which occurring on the Ionian coast and the remaining 17.5% on the
Thyrrenian. Morpho-classes are distributed as follow in the whole population: 32.7% belongs to class 1, 27.7% to class 2, 35.1% to class 3 and 4.5% to class 4. Besides, classes are percentually distributed on Ionian/Thyrrenian stands as follow: class 1 61.2/8.8, class 2 86.7/13.3, class
3 97.2/2.9 and class 4 95.4/4.6. The study considers the ecology and biology of the species together with the human pressure in order to identify areas with high conservation value and proposes concrete measures for the species protection.
P0754 – ePoster
The role of female and hermaphrodite flowers in the gynodioecious–gynomonoecious
Silene littorea
: insights into the phenology of sex expression
Casimiro-Soriguer, I 1,2 , Buide, ML 2 , Narbona, E 2
1 Universidad de Sevilla, Depto de Biología Vegetal y
Ecología, Área de Botánica, Spain; 2 Universidad Pablo de Olavide, Depto de Biología Molecular e Ingeniería
Bioquímica, Spain
Some gynodioecious species showed intermediate individuals that bear female and hermaphrodite flowers; which is considered as a gynodioecious– gynomonoecious sexual system (hereafter GGSS).
Although some works have studied the percentage of different morphs and their reproductive potential in populations with the three morphs, the flowering phenology and specifically the phenology of male and female functions, has not been yet explored. Here, we studied the GGSS of S. littorea Brot. in a phenological context. S. littorea is an annual species endemic to the
Iberian Peninsula. The study was carried out in two natural populations with contrasting habitat during the entire flowering period. Silene littorea showed a GGSS in which most plants in the populations were PMS, and only a very low number were pure females. Most plants in the population transmit their genes equally via pollen and ovules, and plants that were more functionally female did not have a higher female fecundity. Our results indicate that GGSS of S. littorea is plastic as it showed temporal variation across the flowering season.
At the population level, we have found that both functional femaleness and flower production were positively related to temperature and precipitation levels reached during the natural flowering of the species.
Finally, the flowering of hermaphroditic flowers in male and female phases and female flowers showed a similar pattern of flowering, producing that the variation in the estimated mating environment (i.e. relative number of ovules available to pollen which potentially sired these ovules) during the flowering period had a very low fluctuation. We conclude that the female flowers of both female and PMS plants contribute to stabilizing the mating environment over the whole flowering period, and thus can play a role in the maintenance of the GGSS in S. littorea.
P0755 – ePoster
1
Sexual patterns of
Erophaca
flowers, first andromonoic Fabaceae in the Old World
Casimiro-Soriguer, R 1 , Herrera, J 1 , Tallavera, S 1
University of Seville, Depto Biología Vegetal y
Ecologia, Spain
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Our study is focused on Erophaca baetica (old
Astragalus lusitanicus ), a Tertiary relict legume
(subfamily Papilionoideae) that presents two disjunct subspecies in the Mediterranean Basin: subsp. baetica , endemic to the Iberian Peninsula and NW Africa, and subsp. orientalis , endemic to Greece, Cyprus and SW
Asia. This genus, endemic to the Mediterranean Basin, is a long-lived, perennial plant with a woody tuber-like organ (xylopodium) and numerous, fast growing herbaceous stems that start to sprout at the beginning of winter (December), in synchrony with the main rainy season under the Mediterranean climate. Plants up to 2 m in height can be found on shady places, but are more usually around 1 m tall. The stems converge at ground level on the xylopodium, so that individuals can be easily differentiated in the field. From January to April the stems bear clusters of showy white, nectariferous flowers that are visited mainly by bees and bumblebees. We observed with frequency that some Erophaca baetica flowers presented aborted or reduced ovary (they were male functionally). We wanted to quantify the phenomenon and studied the presence/absence of male spatial and temporally, as well as the sexual structure in the individuals. We found an unstable proportion of male/hermaphrodite proportion along the phenology and in the plant structure.
P0756 – ePoster
Occurrence of a member of
Osmundea spectabilis group (Ceramiales, Rhodophyta) in the Laje De
Santos Marine State Park, São Paulo, southeastern
Brazil
Fujii, M 1
Larrea, J 4
, Rocha-Jorge, R
, Cassano, V 5
2 , Barros-Barreto, B 3
, Sentíes, A 4
, Diaz-
, Gil-Rodríguez, M C 6
1 Núcleo de Pesquisa em Ficologia, Instituto de Botânica,
São Paulo, Brazil; 2 Post-Graduate Program
'Biodiversidade Vegetal e Meio Ambiente', Instituto de
Botânica, São Paulo, Brazil; 3 Dept Botânica,
Universidade Federal do Rio de Janeiro, Brazil; 4 Dept
Hidrobiología, Universidad Autónoma Metropolitana,
Iztapalapa, Mexico;
São Paulo, Brazil;
5 Dept Botânica, Universidade de
6 Biología Vegetal (Botánica),
Universidad de la Laguna, Canary Islands, Spain
During the phycological survey in the Laje de Santos
Marine State Park (LSMSP), São Paulo, southeastern
Brazil, a species of Osmundea was found growing in the subtidal zone from 7 to 20 m depth. The LSMP is an area classified as a 'hotspot' for benthic marine algae on the
Brazilian coast and, in the summer, it receives a strong input of South Atlantic Central Waters (SACW), a cold water mass that brings nutrients to the park area, reduces water temperature to 15ºC and forms thermoclines.
Because of this phenomenon it is possible to find in the park some genera that occur mainly in cool waters.
Morphological studies carried out on Osmundea specimens revealed all characters typical of the genus, including the two pericentral cells per each axial segment, and the tetrasporangia cut off from random epidermal cells. The phylogenetic position of this species was inferred by analysis of the chloroplast-encoded rbc L gene sequences from 52 taxa, using three other
Rhodomelacean taxa and one Ceramiaceae as outgroups.
The Brazilian species of Osmundea formed a well-
576 supported clade with the 'Spectabilis group' from Pacific
North American, composed by O. spectabilis, O. blinksii,
O. splendens , and O. sinicola . However, the present specimen diverged in high percentage values from the
'Spectabilis group' representatives (4% –7%), confirming that they constitute a different taxonomic entity. It is the first time that a member of Osmundea related to the
'Spectabilis group' is reported for the South Atlantic
Ocean . Financial support: MICINN – CGL 2010–14881
(Spain), CAPES, CNPq, Fapesp (Brazil), and CBS–
PROMEP (Mexico).
P0757 – ePoster
Morphological and molecular studies on red seaweed
Laurencia oliveirana
(Ceramiales, Rhodophyta): an
1 endemic species from Brazil
Cassano, V
Larrea, J 3
Dept Botânica, Universidade de São Paulo, São Paulo,
Brazil; 2
1 , Gil-Rodríguez, M C
, Oliveira, M C 1
2
, Fujii, M 4
, Sentíes, A 3 , Díaz-
Biología Vegetal (Botánica), Universidad de la
Laguna, Canary Islands, Spain;
Universidad Autónoma Metropolitana, Iztapalapa,
Mexico; 4
3 Dept. Hidrobiología,
Núcleo de Pesquisa em Ficologia, Instituto de
Botânica, São Paulo, Brazil
Morphological and molecular studies were carried out on
Laurencia oliveirana from the type locality (Arraial do
Cabo, Rio de Janeiro, Brazil). This species is easily recognized by its small size, sub erect habit forming intricate cushion-like tufts and unilateral pectinate branching. Lenticular thickenings described as absent were rarely observed for the first time in this species, which displays all characters typical of the genus
Laurencia , such as: the production of the first pericentral cell underneath the basal cell of the trichoblast, two fertile pericentral cells (the third and fourth) without production of additional pericentral cell, spermatangial branches produced from one of two laterals on the suprabasal cell of trichoblasts, and the procarp-bearing segment with five pericentral cells. Details of the procarp and male plants are described for the species for the first time. Molecular analyses using rbc L and cox1 sequences will be presented. The phylogenetic position of L. oliveirana was inferred by analysis of the rbc L gene sequences from 87 taxa, using five other Rhodomelacean taxa and one Gracilariacean as outgroups. In all phylogenetic analyses, L. oliveirana grouped with L. caduciramulosa, L. venusta, L. natalensis and L. flexuosa forming a monophyletic clade within Laurencia sensu stricto. The genetic divergence between L. oliveirana and the molecularly closest species, L. natalensis , was 2.2% .
Financial support: CAPES, CNPq, FAPESP (Brazil),
MICINN - CGL 2010-14881 (Spain), and CBS-
PROMEP (Mexico) .
P0758 – ePoster
Foliar architecture of Burseraceae and
Anacardiaceae
Calvillo-Canadell, L
Millán, M 2
1 , Cevallos-Ferriz, S RS 1 , Martínez-

1 Depto de Paleontología, Instituto de Geología,
Universidad Nacional Autónoma de México, Ciudad
Universitaria, Coyoacan, México; 2 Albert R. Mann
Library, Cornell University, Ithaca, USA
Presence of members of Myrtales in different Cenozoic outcrops in Mexico has suggested its importance in the past vegetation of low latitude North America. The history of this group may be traced to the Upper
Cretaceous of Coahuila, where woods and leaves have been suggested to represent Anacardiaceae. Diverse leaf/leaflets assemblage with impression/compression fossils resembling members of Anacardiaceae,
Burseraceae and Myrtaceae, have been recognized form the Eocene La Carroza Formation, Nuevo León, and the
Coatzingo Formation, Puebla. Recognizing taxa like
Lannea, Cyrtonium, Rhus, Pseudosmodingium,
Comocladia, Haplorhus, Tapirira, Loxopterigium
(Anacardiaceae), Bursera (Burseraceaea) and cf. Myrcia,
Eugenia or Calyptrantes (Myrtaceae) required the revaluation of their leaf architecture patterns. In
Burseraceae some characters that vary among species, may also vary within a single leaf, and in some instances in a single leaflet. For example, the leaf base is asymmetrical, but may also be symmetrical, or apexes are commonly acute, but other types are present. Leaflets are serrate, dentate or crenate, but in many cases these characters do not apply to the whole leaf or leaflet. In some cases the proximal leaf/leaflet zone tends to be entire while towards the distal zone the leaf/leaflet becomes serrate, dentate or crenate; in others, one side is serrate, dentate or crenate while the other is entire. Also the tooth order, from, space between teeth, and number of teeth/cm varies. Secondary venation is a great example of variation in the family. Secondary veins seem to form brochidodromous arches; however, in most cases the arc is formed due to the fusion of adjacent secondaries through a third order vein, giving place to an eucamptodromous pattern. Other patterns based on freely dichotomizing secondaries give place to the cladodromous pattern, however, if one of the branches reaches the sinus of a tooth, while the other joints the super-adjacent secondary, forming an arch, the pattern would be semi-craspedodromous. These four patterns can be found mixed in the same leaf and leaflet. The third order venation is mainly dichotomous, but a percurrent pattern may also be observed. These veins do not fill the intercostal lamina space, but tertiary veins are the ones that actually reach the teeth. The fourth order venation forms either a regular or random reticulum responsible of areole development that includes veinlets, that dicotomize to form fifth and sixth order veins, which may form tracheoid ideoblasts. Similarly, Anacardiaceae does not have a regular pattern that characterizes the family; but some groups may be recognized based on the presence of simple leaves (Anacardieae, Semecarpeae, and Dobineeae) vs compound ones (Rhoeae and
Spondisdeae). The presence of cladodormus venation is also common in this family, where at least 3 special variations can be distinguished. Important is that the brochidodromous condition in Burseraceae is distinct form the one in Burseraceae, being referred as weak brochidodormous. Establishing these differences has allow a more precise identification of the fossil plants, suggesting that their distribution and diversity augmented during the Neogene, possible following the geologic evolution of the area, as other groups suggest.
P0759 – ePoster
Two new species of
Codium
(Bryopsidales,
Chlorophyta) from Australia
Chacana, M 1 , Silva, P 1
1 University of California, Berkeley, USA
Two new species of Codium , C. apiculatum P.C.Silva,
M.E.Chacana et H.B.S.Womersley, and C. unguiculatum
M.E.Chacana et P.C.Silva, are described on the basis of several subtidal collections from southern and southwestern Australia. In the first species the dichotomous branches have acute tips; the utricles are very stout, subcylindrical to slightly clavate, unlike those of any other Australian species. The thallus of the second new species is Colpomenia -like, with the utricles subtended by a thin layer of rhizoidal filaments; the utricles are produced in clusters and have thickened apices resembling fingernails. These two new species, along with several previously unrecorded species encountered in an ongoing survey of Codium in northwestern Australia, attest to the remarkable richness of Australia in terms of the number of species of Codium , which has reached 25 .
P0760 – ePoster
Apogamous ferns with extreme morphological variation: reticulate evolution of
Pteris cadieri complex
Chao, Y-S 1
1
Chiou, W-L 3
, Dong, S-Y 2 , Chiang, Y-C 1 , Liu, H-Y 1 ,
Dept of Biological Sciences, National Sun Yat-Sen
University; 2 South China Botanical Garden, Chinese
Academy of Sciences, China; 3 Division of Botanical
Garden, Taiwan Forestry Research Institute, Taiwan
Apogamous ferns are thought to have limited genetic variation. Because of extreme morphological variation,
Pteris cadieri complex are suggested as hybrid taxa.
However, it is intriguing how and why the apogamous ferns exhibit diverse morphologies. We investigate the evolutionary history of the adieri complex by means of multiple methods. Newly developed primers for nuclear
DNA (PgiC gene, cytosolic phosphoglucose isomerase) were combined with cpDNA markers ( atp Brbc L spacer and rbc L gene) to infer the maternal and biparental lineages of the Pteris cadieri complex. Cytology analysis by chromosome counting and flow cytometry determined diploids, triploids, and tetraploids. Eleven hybrid taxa, with identifiable morphologies, cytotypes, and genotypes, and clear maternal and paternal lineages of each taxon were determined. Furthermore, the genetic variations of Pteris cadieri in two independent areas,
Hainan and Taiwan, denote a multiple origin for this complex. We propose that the Pteris cadieri complex originated from different genetic lineages through multiple hybridizations in different geographical areas, leading to its present morphological diversity.
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P0761 – ePoster
Molecular evolution and genetic diversity of calmodulin gene family in
Oryza
L.
Chao, Y-S 1 , Chiang, Y-C 1
1 Dept of Biological Sciences, National Sun Yat-Sen
University, Taiwan
Rice is an important crop plant and cultivated for several thousand years. Based on adaptation of farming operation, the cultivated rice ( O. sativa ) could be segregated into two groups, lowland rice and upland rice, planted in flooding field and dry field, respectively. Most of cultivated rice is lowland rice, and only 18% of cultivated rice is upland rice. Led by the global climate change, the lack of water resource is severe. In the future, it could be a trend to plant more upland rice because of its drought resistance. Camodulin is a kind of calciumbinding protein and can affect many different cellular functions, including stress response and drought resistance in the previous study. In this study, we focus on the genetic diversity and selective force on the calmodulin gene family of Oryza , including lowland and upland cultivated rice, wild rice, and related species.
There are six copies belong to four chromosomes in
Oryza spp. Three functional copies, in 3rd, 5th, and 12th chromosome, are explored for its genetic diversity in the genus Oryza , especially the molecular evolution in upland rice and lowland rice.
P0762 – ePoster
Identifying potential glacial refugia and postglacial recolonization on the platform of Qinghai–Tibetan
Plateau: a perennial shrub
Spiraea alpina
(Rosaceae)
Chen, S 1 , Zhang, F 1 , Gao, Q 1 , Duan, Y 1 , Li, Y 1 , Fu, P 1
1 Northwest Institute of Plateau Biology, Chinese
Academy of Sciences, China
Spiraea alpina Pall. Fl. Ross. is a perennial shrubby species distibuted on sunny slopes or in rim of shrub widespread across the QTP and the adjacent highlands with an altitude between 2000 and 4500 m. The genus
Spiraea L. has been variously divided by different authors into subgenera, sections, series because of their morphological variation. The present species of Spiraea on the QTP may have migrated from the northeast China by the Early Quaternary or earlier. However, the evolutionary history of Spiraea species during the
Quaternary glaciations is still unknown. Phylogeographic study of Spiraea alpina is useful to obtain a better understanding of the phylogeographic structure of alpine plants on the QTP. By sequencing one intergenic chloroplast spacer, trn Ltrn F (925bp), in 528 individuals from 43 populations sampled throughout the entire distribution range of the species, we found 10 polymorphic sites (1.08%) and identified 10 chloroplast haplotypes (H1¨CH10). The level of differentiation among the populations was relatively high (GST =
0.737). Populations in the RES-QTP (the eastern and southern QTP) were dominated by five haplotypes in which three are ancestral. We suggest that the RES-QTP served as a major refugium for this species through the
578
Quaternary climatic change. Two thirds of all the detected haplotypes occurred in W-QTP (the western
QTP) and 60% of them are unique to this region. Two of three populations in the W-QTP were dominated by 3 haplotypes. We infer that the western region served as another major refugium for this species. Two clearly potential refugia for this species are both on the platform of the QTP: the RES-QTP and W-QTP and this species persisted in these multiple isolated refugia and recolonized to adjacent regions following the glacial retreat. The present wide distribution of the species on the QTP platform has resulted from recent population expansions at least before the last glacial maximum.
P0763 – ePoster
Phylogeny and pre-domestication cultivation in the
Scrophularia ningpoensis
species complex
Chen, C 1 , Li, P 1 , Lee, J 2 , Schaal, B 3 , Fu, C 1
1 Zhejiang University, Hangzhou, China; 2 Research
Institute of Bioscience and Biotechnology,Daejeon,
Korea; 3 Washington University, St Louis, USA
Scrophularia ningpoensis Hemsley (Scrophulariaceae), known as Zhexuanshen, is used in the Chinese Materia
Medica (CMM) and has a long history of widespread use in China. It is used to treat inflammation, laryngitis, tonsillitis, abscesses and constipation. Recent research revealed that this medicinal species, which has high antiangiogenic activity, also can be used as an anticancer agent. S. ningpoensis, which is endemic to China, was named by Hemsley (1890) based on specimens collected in Tiantong, Ningbo of Zhejiang. The species now is widely cultivated in China but over the past several decades, wild populations of S. ningpoensis have suffered rapid decline because of over-exploitation.
Based on our phylogenetic analyses of the East Asian
Scrophularia using the nuclear ribosomal internal transcribed spacer (nrITS), S. ningpoensis groups with S. buergeriana, S. kakedensis and S. yoshimurae into a single clade which is monophyletic and sister to other species of Scrophularia . It is clear that these four species, found in China, Korea and Japan, could be considered a species complex, and thus have been chosen to further examine the domestication of S. ningpoensis . Four species, with a total 40 populations were surveyed for chloroplast DNA variation in the spacer regions trn L-F and pab Atrn H. Results of the cpDNA analysis showed lower genetic diversity in cultivated populations than in the wild populations. Native S. ningpoensis populations have 20 distinct haplotypes, whereas cultivated populations contain only four haplotypes in total. Based on a cpDNA haplotype network, geo-authentic cultivated populations in Zhejiang are not introduced from local wild populations. Contrary to current belief, they are all from the Jiangxi wild population. The highest diversity of haplotypes within populations is found in the Tianmu
Mountains, which are considered a glacial refuge for many plant species. All haplotypes constituted a gene tree without obvious separate clusters for cultivated and wild populations, suggesting multiple origins of cultivated S. ningpoensis .

P0764 – ePoster
Genetic diversity of the medical parasitic plant
Cynomorium
Chen, GL 1 , Liu, GD 1 , Feng, D 1 , Chen, Q 1
1 Inner Mongolia University, Huhhot, P.R. China
Cynomorium has two species in eastern Asia
( Cynomorium songaricum Rupr.) and the Mediterranean
( Cynomorium coccineum ). Cynomorium songaricum is used as an important medicine in China. For the better use of Cynomorium songaricum Rupr., HPLC fingerprinting and Inter-Simple Sequence Repeat (ISSR) molecular markers were carried out, involving 16
Cynomorium songaricum populations from different locations in China. We selected 7 UBC Primers for genetic diversity analysis, a total of 102 bands were generated, of which 77 bands were polymorphic band
(the percentage of polymorphic band PPB = 84.8%). Nei¡ similarity coefficients was 0.2623, Shannon's
Information index was 0.3913 and genetic similarity coefficient was changed from 0.1942 to 0.5141. The results indicated that there actually existed much genetic diversity at molecular level among different populations of Cynomorium songaricum . Clustering analysis of species of the 16 populations was also performed based on the variation patterns of 14 chemical constituents using UPGMA method. This study demonstrated that the
UPGMA dendrograms of the 16 Cynomorium songaricum populations based on their chromatograms of chemical components and molecular UPGMA dendrogram were not matched perfectly with one another. Genetic diversity was closely related to geographic location, 8 populations from Inner Mongolia autonomous region have higher genetic similarity.
Chemical constituents’ diversity was related to the environments in some way, so specimens of
Cynomorium songaricum which were collected from the same region and grew in similar enviroment had the higher similarity. Cynomorium songaricum collected from Hangjinqi was clustered to a special chemotype for its special enviroment.This research also shows that combining the use of genetic base with chemotype will help control the favourable chemotypes and breed new cultivars with more desirable chemical constituents.
P0765 – ePoster
A preliminarily taxonomic revision on the genus
Bougainvillea
Comm. (Nyctaginaceae)
Chen, T 1 , Nee, M 2
1 Shenzhen Fairy Lake Botanical Garden, China; 2 New
York Botanical Garden, USA
12 species have been preliminarily recognized among more than 30 species names proposed under the genus
Bougainvillea based on investigations of specimens and images of main herbaria in the world. These species native to South America are distributed in the dry tropical and subtropical caatinga and chaco and vicinities in Argentina, Bolivia, Brazil, Paraguay and Peru. Plants of the genus occur in other countries as cultivars or rarely introduced species. The status of the species B. buttiana or hybrid B.
x buttiana is doubted. The comprehensive taxonomic revision on Bougainvillea is laying a base for clarifying the pedigree of about 350 cultivars evolved within the genus.
P0766 – ePoster
The study and monitoring of an urban forest dynamics plot in Taiwan – assessment of a botanic garden site
Chiu, S-T 1 , Li, W-C 2
1 Dept of Botany, National Museum of Natural Science,
Taichung; 2 Dept of Ecology, Providence University, USA
An urban dynamic plot at Reijin, Dadu, Taichung was established for the purposes of research, education, conservation and monitoring the mid-west vegetation at low elevation in Taiwan. The aims of this study were to investigate the native species and the plant community by an inventory of the area in the rather preserved forest, to carry on further studies, and also to provide the foundation of long-term monitoring the urban forest succession. A total of 153 species in 133 genera of 63 families were recorded in 78 plots (20 m – 10 m) of 1.56 ha area; including all the trees and shrubs with DBH>1 cm, the lianas with DBH>0.5 cm, and the plants of ground coverage from August 2007 to November 2009.
Among these, a total of 4136 stand woody plant individuals were recorded in 50 species of 48 genera of
25 families. The density was 3105 stems/ha and total basal area of 130 cm above ground were 15.51 m 2 /ha.
The most dominant canopy species were Acacia confusa
Merr. with a bell-shaped pattern in the analysis of the size-class structure of woody plants. The dominant subcanopy and shrub species were Mallotus paniculatus
(Lam.) Muell.Arg., Tricalysia dubia (Lindl.) Ohwi,
Litsea hypophaea Hayata. A total of 2029 liana stems were recorded in 20 species of 20 genera in 14 families as 1406 stems/ha in density and 4296.07 cm 2 /ha for total basal area. The most dominant species were
Trachelospermum jasminoides (Lindl.) Lem., Morinda parvifolia Bartl. and Gymnema sylvestre (Retz.)Schult, and the size-class distribution of them was an L-shaped pattern. The species of ground coverage were recorded
139 species of 120 genera in 64 families. The most dominant species were the invasive Panicum maximum
Jacq., native Microlepia speluncae (L.) Moore and native
Cyrtococcum patens (L.) A.Camus. The result of analysis which combines CCA and DCA showed the important roles of the slope, soil moisture and soil-EC variables in the trees and shrubs and of the slope, soil-pH variables in ground coverage. This is the first essential and broad study of urban dynamic plots in Taiwan. We found the remnants of rare species, the high diversity of original native plant species, the good recruitment of population in Acacia confuse secondary forest and also the invasive species in progress. It is not only providing a foundation for integrating the historical development of intensive agriculture and forestry urbanisation and other land use changes, but also initiates a program for garden safeguarding local plant diversity, monitoring vegetation succession, developing environment education project and implementing an in-situ and ex-situ conservation of the sustainable use of plant genetic resources. The perspective of the urban dynamic plot of the Botanic
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Garden can become a plant-community-based approach to monitoring the under threat vegetation and rare species conservation.
P0767 – ePoster
Phylogeography and genetic diversity of east Asian
Neolitsea sericea
(Lauraceae) using chloroplast DNA sequence variation
Choi, B H 1 , Lee, J H 1
1 Dept of Biological Sciences, Inha University, Republic of Korea
Neolitsea sericea (Lauraceae) is an evergreen broadleaved tree growing in East Asian warm-temperate regions. The species is a common plant of evergreen forests in Korea and Japan, and shows a disjunctive distribution on the Lanyu Island, Taiwan and the
Zhoushan Archipelago, China. We attempt to reconstruct the phylogeographic history of this species and to infer the refugia of warm-temperate evergreen broad-leaved plants in the Far East during the Quaternary ice age. We identified the chloroplast DNA haplotypes of 325 individuals from 36 populations covering entire geographic range of the species based on sequence data from trn L-F and psb Ctrn S intergenic spacers and rps 16 intron. The nine haplotypes were identified and the haplotype network constructed using the program TCS.
To examine the genetic structure an analysis of molecular variance (AMOVA) was also carried out. No dominant haplotype was widespread over the entire distributional range of this species. Most of the haplotypes were revealed in Chukoku region of Japan, so that the region is supposed to be the refugia during the climatic oscillations. Rare haplotypes are found in Korea and
Lanyu Island respectively. Two haplotypes detected in
Zhoushan Archipelago are common to the populations of
Korea and Japan. Our results indicated that the present haplotype distribution of the species has resulted from multiple migrations through land bridges connecting
Japan to the Asian continent or Taiwan during the glacial periods and the isolations of populations have occurred according to sea level rising after the ice age.
P0768 – ePoster
Chromosomal mapping of 18S-25S and 5S ribosomal genes of Fagaceae species from northern Thailand
Anamthawat-Jónsson, K 1 , Chokchaichamnankit, P 2 ,
1 Faculty of Life and Environmental Sciences, University
2 of Iceland, Reykjavik, Iceland; Faculty of Science,
Chulalongkorn University, Bangkok, Thailand
Fagaceae (beech family) includes 7–12 genera and 600–
1000 species distributed worldwide. In Thailand, this family comprises four genera: Castanopsis (D.Don)
Spach. (chestnut, mostly evergreen, 33 species);
Lithocarpus Blume (stone oak, mostly evergreen, 56 species); Quercus L. (oak, mostly deciduous, 29 species); and Trigonobalanus Forman (evergreen, one species). In this study, sixteen species were investigated: eight
Castanopsis , four Lithocarpus , three Quercus species and Trigonobalanus doichangensis . The plants under
580 study were from Khun Mae Kuong Forest in Doi Saket district, Chiang Mai province, at approximately 18.87˚N
/99.14˚E, northern Thailand. All samples were collected in the field, from six locations, representing three types of habitats, i.e. hill-evergreen forest and hill-evergreen forest with pine, both at relatively high elevation (1000–
1800 m), and dry-deciduous forest at altitudes below 800 m. At each location, twenty trees or more, 5–30 m tall and 30–100 m apart, were randomly selected, marked with identification number, and local names recorded together with their position/map within the forest. Leaf buds, and flower buds if available, from individual trees were collected for chromosome isolation. Twigs with fully grown leaves, fruits (acorns) and flowers were collected for taxonomic identification and kept as voucher specimens. The species belonging to
Castanopsis, Lithocarpus and Quercus were found to be generally diploid with the chromosome number 2n = 24, and the basic number x = 12 was confirmed in a number of species with meiosis. On the other hand, T. doichangensis was found, by both mitotic and meiotic analyses, to be diploid with 2n = 14 and x = 7.
Chromosomal mapping of the highly repetitive 18S-25S and 5S ribosomal genes by fluorescence in situ hybridisation (FISH) was performed. Most species (from all four genera) showed four 18S-25S rDNA sites (two pairs: one sub-telomeric major and one paracentromeric/intercalary minor loci) and two 5S rDNA sites (one pair: para-centromeric locus). Some species showed other numbers and locations of ribosomal sites, for example both pairs of the major 18S-25S rDNA loci in Q. kerrii were sub-telomeric, whereas C. argentea and
Q. brandisianus only had one pair of 18S-25S rDNA sites. Two species, C. calathiformis and L. vestitus , showed an odd number of (unpaired) sites, and this indicated hybrid origin and/or polyploidy. Polyploid cells were detected in root-tip samples of these species. In general, the ribosomal gene maps based on both sequences together can be used to support the taxonomic classification, at least at the genus level. In Castanopsis , the 18S-25S and the 5S genes were localized on three different chromosome pairs, and comprised speciesspecific maps. On the other hand, the ribosomal genes in
Lithocarpus and Quercus were found only on two chromosome pairs, because one of the two 18S-25S rDNA loci was localized on the same chromosome as the
5S rDNA locus. Based on this ribosomal FISH mapping,
T. doichangensis appears to be closely related to the oak genera Lithocarpus and Quercus . The FISH markers may be used to clarify discrepancies arising from morphological assessments.
P0769 – ePoster
Sahul origin and multiple long-distance dispersals of
South Pacific alpine plants: insights from molecular dating, phylogeography and population genetics of
Oreomyrrhis
clade of
Chaerophyllum
Chung, K-F 1 , Peng, C-I 2 , Schaal, BA 3
1 School of Forestry and Resource Conservation,
National Taiwan University;
Center, Academia Sinica; 3
2 Biodiversity Research
Dept of Biology, Washington
University, USA

The Oreomyrrhis clade of Chaerophyllum is distributed across the alpine, subalpine, and sub-Antarctic habitats around the South Pacific Basin (i.e. Central America, northern to central Andes, Tierra del Fuego, the
Falklands, New Zealand, SE Australia, Tasmania, New
Guinea, Borneo, and Taiwan). Though its main distribution aligns well with the South Pacific Gondwana landmasses, recent phylogenetic studies indicates a relative young (Pliocene–Pleistocene) age of its diversification, supporting the scenario of transoceanic long-distance dispersal (LDD) for its remarkable disjunction. However, patterns of LDD of this young clade remain unsettled due to extremely low phylogenetic resolution. To unveil the intriguing historical biogeography of the Oreomyrrhis clade, a combination of molecular dating, phylogeography and population genetics was conducted. Phylogenetic analyses of the combined dataset (nuclear ITS and chloroplast trn S -trn G and atp B -rbc L sequences) reveal two clades lacking internal resolution: a clade composed of New Guinean and Bornean species and another clade comprising all other species. Molecular dating supports
New Guinea as the most likely center of origin as the
Oreomyrrhis clade has diversified for the longest there, followed by the Australian species. Star-like haplotype networks and significant negative values of Tajima's D test for all three datasets imply a biogeographic scenario dominated by range expansion. In all three haplotype networks, only the most ancestral haplotypes are found in more than one area, suggesting that dispersals from the center of origin had happened prior to the origination of more derived haplotypes. Assuming that the center of origin is the region harboring the ancestral haplotypes and the highest genetic diversities, Australia stands out as the most likely birthplace for the Oreomyrrhis clade, followed by New Guinea. In the atp B -rbc L network, the ancestral haplotype and an interior haplotype adjacent to it are found in multiple regions, suggesting that initial range expansions into Taiwan, New Guinea/Australia, and the New World each involved at least two independent dispersals. Results of these analyses suggest either Australia or New Guinea is the most likely center of origin for the Oreomyrrhis clade. However, when glacial histories of the region are taken into account, this competition can be reconciled. As New Guinea was connected to Australia to form the super continent Sahul during the glacial maximum, we propose a Sahul origin and diversification of the Oreomyrrhis clade, followed by
LDD to Taiwan, New Zealand and America.
P0770 – ePoster
Specialisation vs. generalization – field investigations in bee-pollinated
Salvia
(Lamiaceae) species from
Central Atolia
Celep, F 1 , Atalay, Z 1
Bockhoff, R 3
, Dikmen, F 2 , Dogan, M 1 , Claßen-
1 Dept of Biological Sciences, Middle East Technical
2 University, Ankara, Turkey; Dept of Biology, Hacettepe
University, Ankara, Turkey; 3 Institut für Spezielle
Botanik, Johannes Gutenberg Universität, Mainz,
Germany
Bilabiate flowers evolved in co-evolution with bees and thus represent a classical example for specialisation to a certain pollinator guild . This is also true for Salvia , the largest genus in the Lamiaceae with approximately 680 bee-pollinated species. In view of the huge number of sages and bees we question how close the mutualism between a given flower and its bees might be. In our study, we investigated nine co-occurring Salvia species
( S. absconditiflora Greuter & Burdet, S. aethiopis L., S. candidissima Vahl, S. cyanescens Boiss. & Balansa, S. hypargeia Fisch. & Mey., S. russellii Benth., S. tchihatcheffii Boiss., S. virgata Ait., S. viridis L.), one hybrid ( S. candidissima x S. cyanescens ) and their pollinators in central Anatolia, Turkey. We identified 32 insect pollinators, among them 28 bee species from 15 genera. They either preferred only a single Salvia species
(18 bee species) or visited up to seven different sages
(e.g. Apis mellifera ). Then, specific pollination was guaranteed by seasonal and/or mechanical isolation due to differential pollen deposition by means of the staminal lever mechanism. The only exceptions were the hybridizing S. candidissima and S. cyanescens (sharing six pollinators) and the not hybridizing S. viridis/S. virgata (sharing Anthophora aestivalis ) and S. absconditiflora/S. tchihatcheffii (sharing Apis mellifera and Osmia mustelin a) flowering simultaneously and depositing pollen on the same site on the pollinator. For the first time, fly pollination was documented in a Salvia species as Nemestrinus caucasicus Fischer, N. reticulatus
Latreille, Neorhynchocephalus tauscheri Fischer (all
Nemestrinidae) and Pangonius pyritosus Loew
(Tabanidae) were observed as frequent pollinators in S. virgata sharing the niche with 21 different bee species.
The nine Salvia species and the hybrid usually share 3–7 pollinators (7 ssp.) with only two pollinators observed in the hybrid and 12 and 25 pollinator species in S. russellii and S. virgata , respectively. In each case, the pollinators belonged to different genera and clearly differed in body size and proboscis length. Though they appeared in different frequencies per species, they clearly indicated that the Salvia species investigated had almost no specialization to a certain bee species, bee subgroup or bee size. We conclude that in our model system the bees are much more specialized than the Salvia species which indeed appear to be functional generalists within the limits of their bilabiate flower construction.
P0771 – ePoster
Who is who in
Salvia
L.? A phylogenetic update of the sages from the Old World (Lamiaceae)
Will, M 1 , Claßen-Bockhoff, R 1
1 Institut für Spezielle Botanik, Johannes Gutenberg-
Universität, Mainz, Germany
Phylogenetic reconstruction of the relationships within the genus Salvia L. (Lamiaceae, approx. 900 spp.) has been largely restricted to the New World. Until now, only 27 (7.9%) species of the European, African,
Southwest and Central Asian sages have been investigated although the origin of the genus might be located in this area. In addition, the unresolved position of the Old World branches calls for further investigation.
Based on nuclear (ITS) and chloroplast (rpl32trn L) sequence data we introduce the first subgeneric classification of Salvia with a special focus on the circum-Mediterranean species. We covered the whole
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distribution area of the genus with a broad geographical sampling and generated ITS sequences for 139 taxa from the Old World that were never sequenced before.
Therefore the Old World is now represented by 52.3% of the recognized species and we hope to expand the sampling to cover 80% (without Central Asian sages).
Generally, our data support two of the three major clades
(Clade I & II) defined by Walker & Sytsma (2007) as well as the monophyly of two additional Old World clades (Central Asian and South West Asia). We achieved a better resolution and a slightly different topology concerning the placement of Rosmarinus and
Perovskia . Furthermore, we defined some highly supported monophyletic lineages within the circum-
Mediterranean Clade I. One of them comprises 16 South and East African species and two taxa from Madagascar.
Interestingly, we found several independent dispersal events to South (2x) and North (5x) Africa out of the
East Mediterranean area (Turkey). All European endemics are placed in Clade I as well. Only the Chinese and Iranian representatives of the genus form two independent clades. Within the latter, we detected an additional dispersal to Northern Africa and the Canary
Islands rendering the Canarian species polyphyletic.
Furthermore, we analyzed rpl32trn L sequence data. The comparison of nuclear and chloroplast datasets revealed incongruence that might indicate a hybrid origin of the
Mediterranean clade and an ancient allopolyploidization.
These findings gain additional support from morphology
(floral traits) and caryology. Finally, our data support a sister relationship of Meriandra and Dorystaechas and a monophyletic clade consisting of Perovskia, Melissa and
Lepechinia , the latter in an unresolved position close to or within Salvia . Based on a combined dataset (ITS and cp-DNA sequences) we intend to resolve the backbone of the topology and to identify the geographical origin of the genus. This would allow for the identification of evolutionary tendencies, i.e. the modification of the characteristic lever (stamen type) and of floral traits in adaptation to local pollinator guilds, respectively.
P0772 – ePoster
Diversity and conservation of bromeliads of the Vale do Ribeira, State of São Paulo, Brazil
Coffani-Nunes, J V 1
1 ICBS, UFAL, Maceio, AL, Brazil
Bromeliaceae is the largest almost exclusively tropical family among the angiosperms. Its 3172 species are presently organized in 58 genera and eight subfamilies.
The two largest centers of diversity in the family are eastern Brazil and the Tepui region in Venezuela. Brazil contains a large number of bromeliad species; it is estimated that about 70% of the genera and 40% of the species occur in Brazil, especially in the southeast where the Atlantic Forest biome predominates. The Atlantic
Forest is one of the hotspots of highest biodiversity on the planet and is the ecosystem that suffered most from the socioeconomic development of the country. In the remnants of this ecosystem occur several endemic genera of Bromeliaceae, such as Canistrum, Fernseea, Lymania,
Nidularium, Portea , and Quesnelia . The Vale do Ribeira, located in the southern part of the State of São Paulo, is contained completely within the Atlantic Domain and
582 concentrates the largest remnant of the Atlantic Forest in the State of São Paulo and one of the largest in Brazil.
The compilation of a database on the vegetation of the
Vale do Ribeira, resulted in the publication of the ‘Flora do Vale do Ribeira: listagem de Angiospermas’. Based on this database, the information on Bromeliaceae was combined with the data from the ‘Flora Fanerogâmica do
Estado de São Paulo’ and ‘Plantas da Mata Atlântica’ to produce a definitive list of the species richness and endemic species of the Atlantic Forest. With respect to the conservation status, the classification from the ‘Livro
Vermelho das Espécies Vegetais Ameaçadas do Estado de São Paulo’ was adopted. The abbreviations for authors of the species were verified through the site of the ‘Lista de Espécies da Flora Brasileira’ and from the ‘Flora
Fanerogâmica do Estado de São Paulo’, with some added considerations. In the Vale do Ribeira, 75 species belonging to 14 genera occur, corresponding to 45.73% of the species of Bromeliaceae of the State of São Paulo.
The most important genera were Vriesea (27 spp.),
Aechmea (10 spp.), and Nidularium (10 spp.), that is,
62.66% of the richness of the region. More than half of the species (61.33%) are endemic to the Atlantic Forest biome. In addition to the endemic genera, Vriesea and
Aechmea show a high rate of endemism for the biome,
77.77% and 50% of their species, respectively. The great majority of species do not present serious conservation problems; only five are classed as Vulnerable (VU) and two as Nearly Threatened (‘Quase Ameaçadas’, QA).
Also as a result of this study, two additional species are indicated, Quesnelia violacea and Vriesea flava , for inclusion in the list as Vulnerable. The present study demonstrates the importance of the Vale do Ribeira in the preservation of bromeliads of the Atlantic Forest. It is expected that, as research continues, new occurrences will be reported, since many of the areas where native vegetation still remains have not yet been inventoried.
P0774 – ePoster
Molecular phylogeny of
Haworthia
(Asphodelaceae): evidence from plastid and nuclear DNA sequences
Daru, B H 1 , Maurin, O 1 , Van Der Bank, M 1
1 African Center for DNA Barcoding, University of
Johannesburg, South Africa
Haworthia Duval (Asparagales: Asphodelaceae:
Alooideae) is endemic to southern Africa with modifications to withstand relatively waterless or desert environments. Despite its economic importance in horticulture, its molecular systematics remains unresolved. Haworthia comprises 3 subgenera. Species delimitation and placement within one of three subgenera was based on flower character and leaf shape. Because a complete phylogeny of the group was not available, hypotheses regarding species definition were not tested.
This study addresses generic delimitation of Haworthia using molecular sequenced data from three plastid
( rbc La, mat K and trnH-psbA) and the first subunit of the nuclear ribosomal internal transcribed spacer (ITS1) gene regions. Representatives of other genera within
Alooideae were also included to infer the placement of the three subgenera of Haworthia within Alooideae. The findings are discussed.

P0775 – ePoster
Traditional dye yielding plants of West Bengal, India
Das, PK 1 , Mondal, AK 1
1 Vidyasagar University, India
The present paper based on extensive field survey, collection of the original knowledge on dye preparation and various ethno-botanical information of Paschim
Medinipur district, West Bengal, India. About 22 important dye yielding plant materials have been collected and properly identified and simultaneously plants were preserved in the Departmental Herbarium for future information along with their vernacular name, habit, part(s) used, nature of dye, medicinal and economic values have also been recorded. Research has shown that the natural dyes are environment friendly, but we should know the proper utilization of natural dye which is diminished due to lack of knowledge and also easy availability of synthetic dyes. These natural products need proper documentation and conservation for future otherwise we are supposed to lose valuable data for ever .
P0777 – Poster
Biogeodiversity research for characterizing habitatstructure and landscape-types of the Great
Hungarian Plain in southeast Hungary with the use of habitat-mapping
Deák, ÁJ 1 , Nagy, E 2
1 University of Szeged, Dept of Physical Geography and
Geoinfornatics, Hungary; 2 University of Pannonia,
Georgikon Faculty, Dept of Crop Production and Soil
Science, Hungary
The aim of this research was to reveal the habitat-pattern of the Great Hungarian Plain situated in Southeast
Hungary considering the interactions between abiotic and biotic factors. The habitats were ordered into microregionally specific, successionally and dynamically connective habitat-complexes and more comprehensive vegetational landscape-types and main-types to specify the border of geographical microregions and flora districts of Praematricum and Crisicum. The landscapelevel vegetation pattern was mapped in Csongrád county
(3000 km 2 ) with the help of Hungarian Habitat Map
Database using 35 ha-sized hexagonal grid-based which resulted rasteric habitat maps (first author mapped 70% of it on field). The local habitat-pattern was studied on smaller sample areas mainly in Natura 2000 sites with the help of polygonal habitat maps. The habitats were classified according to the ÁNÉR (General National
Habitat-classification System). In sand landscapes the habitat-complexes of the mosaics of deflation hollows with fen-head – alkali foot pattern, steppe grasslands and forests (typical for Dorozsma-Majsaian Sandlands) and the fields of blow-out dunes covered with open sand grasslands and forests (typical for the mid-ridge of the
Kiskunságian Sandlands) were separated. ‘Pusztas’ of loess-landscapes include the habitat-complexes of the mosaics of loess steppe grasslands and oak loess-steppe forests, the wet alkali habitats and the salt-berm alkali steppes. As their typical habitats (loess steppegrasslands, Artemisia alkali steppes and Alopecurus dominant salt meadows) appear in the eastern part of
Danube–Tisza Interfluve the Kiskunságian Loesslands,
Szegedian Plain and Gerje-Perje Plain belong to
Crisicum flora district. 5 vegetational landscape-types were identified in the floodplains which are the active floodplain (active floodplain’s oxbow-lakes; navvyholes; grassland, forest, arable land and orchard mosaics of active floodplain; river-banks; dyke-vegetation); the saved-side non-saline low floodplain (saved-side oxbow lakes; mosaics of non-saline grasslands, swamps and forests; saved-side low floodplains with channels, boundaries and arable lands); the floodplain moorlandscape (mixed eutrophic and moor-lake wetlands at loessland-border; mosaics of moor and floodplain habitats at sandland-border); the saved-side low floodplain secondary saline landscape ( Achillea and meadow-steppe subtypes); and the saved-side lagsurfaces of high floodplains (sand and loess lagsurfaces). The floodplain moor-landscape appears just in the South-Tisza Valley. The active floodplain’s meadows remained mainly in the Hármas-Körös Floodplain and in the upper part of Lower Maros Floodplain as a result of landscape use changes. The eutrophic reed-grasses,
Butomus, Eleocharis, Alisma, Oenanthe dominated swamps appear mainly alongside the river Tisza and
Hármas-Körös, while the annual wet pioneer vegetation, willow-shrubs and hard-wooden alluvial forests in the
Lower Maros Valley. Secondary saline landscapes developed mainly in the South-Tisza Valley and in the
Hármas-Körös Floodplain. The co-presence of non-saline floodplain vegetation and the loess-steppe and salt-berm vegetation covered lag-surfaces separates the Körösszög and Bánságsarok from the other floodplain microregions.
As a result of the diverse abiotic features the Körösszög has the highest habitat-diversity among the microregions of Csongrád county. The habitat-diversity hot spots are at the borders of geographical microregions, in the midthird of the Dorozsma-Majsaian Sandlands
(groundwater-upwelling zone with deflation hollows covered by fen and alkali vegetation), at the salt-berm alkali steppes and in the floodplains (especially in the active floodplain).
P0778 – ePoster
A study of apple cultivation practices and mitigation of various diseases by ethnobotanical methods in
Ganga Valley of Uttarkashi district (Uttarakhand)
Dhingra, G K 1 , Arya, R 1 , Kumar, S 1 , Rather, M A 2
1 R.C.U Govt. P.G.College Uttarkashi, Uttarakhand,
India; 2 Dept of Chemistry RCU Govt PG College
Uttarkashi, Uttarakhand, India
Apple is commercially the most important temperate fruit and is fourth among the most widely produced fruits in the world. About 99 percent of India’s apple area falls under North Western Hills Region, covering 6 districts of
Jammu & Kashmir, 6 districts of Himachal Pradesh and
8 districts of Uttarakhand. The study area is concentrated around the Ganga valley regions (Sukhi, Jhala, Jaspur,
Harsil and Dharali) of Bhatwari tehsil located in
Uttarkashi district of Uttarakhand state. The purpose of this research work is to describe the growth, habits,
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identification of various apple varieties and disease pests.
The geographical & climatic conditions of Uttarakhand are suitable for apple production. But number of diseases, adverse weather conditions like inadequate Chilling
Units Accumulation, temperature fluctuation, inadequate rainfall, hail storms during flowering & fruit development many times hamper the productivity of apples leading to financial insecurity among the apple growers of Uttarakhand. In research sites, apple varieties produced are mainly Red, Brown and Golden Delicious,
Yellow Golden, Golden, Royal Delicious, and Wilsan variety, etc. Apple trees are prone to several pests and diseases depending on our local conditions. The key to healthy apple trees is prevention, and prevention consists of cleanliness – remove and burn any fruit which shows signs of disease, regularly sweep up and burn fallen leaves, and keep the storage area clean with an annual wash of soda and warm water. In this way, the use of chemicals can be avoided. During this study various diseases, viz. grey powdery coating on leaves and young shoots, ribbon-like scars on the apple skin, holes in the leaves, white fluffy areas on the bark, looks like fungus, sunken, discolored patches on bark, small brown sunken areas on the skin of the fruit, bBrowning of the fruit, especially those in storage, brown blotches on leaves and fruit were also reported. Local people of study area mitigate diseases on apple plants by ethnobotanical methods.
P0779 – ePoster
Interactive digital key for the frailejones (Espeletiinae
Cuatrec., Asteraceae) of the South American páramos
Diazgranados, M 1, 2 , Rodríguez, N 3 , Barber, J 0
1 Saint Louis University, Dept of Biology, St Louis, MO,
USA;
USA; 3
2 Missouri Botanical Garden, Saint Louis, MO,
Universidad de los Andes, Bogotá DC, Colombia
The páramos of the Northern Andes are largely dominated by species known as frailejones, belonging to the subtribe Espeletiinae (Asteraceae). The subtribe is a neotropical monophyletic group, distributed from 1,800 m in elevation up to the lower limit of the glaciers (4,700 m) in Colombia (ca. 81 spp.), Venezuela (ca. 69 spp.) and Ecuador (1 sp.). The group is recognized by numerous synapomorphies including a characteristic rosette habit (sometimes caulescent or arboreous) with spiral leaf phyllotaxis; obpyramidal to prismatic shape of the epappose achenes; fertile female ray flowers and functionally male disc flowers; pluriseriate involucre and persistent pales of the receptacle; thick and woody stems; xeromorphic structure; and a static chromosome number
(n=19). The group has been considered a classic example of rapid radiation in the tropics due to its remarkable diversity, which appears to have evolved over a relatively short period of time (< 2-4 my BP). The 143 named species are grouped into eight genera: Carramboa (5 spp.), Coespeletia (6 spp.), Espeletia (68 spp.),
Espeletiopsis (27 spp.), Libanothamnus (11 spp.),
Paramiflos (1 sp.), Ruilopezia (24 spp.) and Tamania (1 sp.). Unfortunately, only one recent key (published in
1996) exists for a single genus ( Espeletiopsis ), as well as one extremely incomplete treatment for Espeletia published in 1935 that comprises only 20% of the currently recognized species. Cuatrecasas’ unpublished
584 monograph of the subtribe does not include more than 15 recently discovered species and describes only seven of the eight genera (excluding Espeletiopsis ); furthermore, its dichotomous keys are particularly difficult to follow due to the large number of overlapping continuous characters. We constructed an interactive digital key for the Espeletiinae, using the software LucID3 v.3.4. The key includes all 143 currently known species and incorporates 566 features (=character states). As implemented in LucID3, features of multiple-state variables were scored using seven states (absent, common, rare, uncertain, common and misinterpreted, rare and misinterpreted, and not scoped), whereas five states were used for numeric variables (absent, normal, uncertain, misinterpreted, not scoped), in addition to numeric normal ranges and extreme values. Numerous problems were encountered, primarily due to a lack of consistency between descriptions, such as the use of 37 different adjectives to describe leaf shape. After solving these problems, often through the re-examination of specimens, the performance of the key was tested. On average, 69 features can be used to distinguish any species from a random pair of species, and the most similar species can be distinguished by 23 features. The key includes photographs and illustrations for all species and for details of morphological characters, illustrated glossaries (e.g. plates with leaf shapes or trichomes), photographic galleries and distribution maps. It is available online and shortly will be available on DVD.
P0780 – ePoster
1
Linnaeus and the flora of the Guianas
Dorr, L 1
Smithsonian Institution, USA
The Guianas figured prominently in the life of Linnaeus.
In 1737, he was offered the post of physician with the
Dutch West India Company but declined and recommended Johann Bartsch. Bartsch was murdered six months after assuming the post in Suriname and while
Linnaeus expressed remorse at the death of his friend, his own travels in the more salubrious Europe established the course of his career. Linnaeus drew on an extensive literature to inform his taxonomy of South American plants, yet the immediate sources of his knowledge of the flora of the Guianas came from a relatively small set of specimens, books, manuscripts, and correspondence.
Bartsch is cited several times in the Species plantarum
(1753); the information abstracted from correspondence.
Maria Sibylla Merian’s Metamophosis insectorum
Surinamensium (1705) also is cited in the same work; the information derived from the 60 copperplates in her book that show insect food plants. In 1755–56, Linnaeus sent
Daniel Rolander to collect in Suriname, but the two had a falling out and Rolander sold his manuscripts and collections to Christian Friis Rottböll. In 1770, Linnaeus received a brief manuscript and several specimens that
Frédéric Louis Allamand collected in Suriname, and in
1774, Linnaeus received a gift of Suriname plants from
King Gustav III. The plants, many preserved in alcohol, were collected by Carl Gustaf Dahlberg and are the basis of the dissertation Plantae Surinamenses (1775). French
Guiana is scarcely mentioned in Linnaeus’s publications.
Pierre Barrère’s Essai sur l’Histoire naturelle de la

France équinoxiale (1741) is noted, but there are no specimens from Barrère’s three years in Cayenne.
Venezuela, however, is frequently mentioned. Pehr
Loefling, a Linnaean disciple, was attached to a Spanish expedition sent to South America to survey the disputed boundary between Spain and Portugal. He arrived in
Cumaná and eventually traveled to the Río Caroni in the
Venezuelan Guayana where he died. Linnaeus cobbled together information on the plants of this region taken from correspondence and manuscripts salvaged and returned to Europe, and Linnaeus published it posthumously as Loefling’s Iter Hispanicum (1758). In at least one instance it appears that Linnaeus tied a
Rolander specimen from Suriname to one of Loefling’s
(Linnaeus’s?) descriptions in the Iter Hispanicum .
P0781 – Poster
Norantea
s.str. (Marcgraviaceae) – taxonomy and biogeography
, Bumler, V 1
1
Dressler, S 1
Senckenberg Research Institute, Botany, Frankfurt/M.,
Germany
Norantea is a genus of lianas flowering in the canopy of neotropical rain forests. With their conspicuous racemose inflorescences including extrafloral nectaries the members attract many visitors and form a valuable food source in these ecosystems. Formerly comprising some
45 species (s.l.) the new classification of the family treats four morphological types as genera, Norantea (s.str.) being one of them with only 3 taxa. We give an overview about the taxa, their morphological characters, ecological traits, and distributional range. Furthermore, models of the possible distribution ranges using Maxent will be discussed. Norantea guianensis is a species of lowland rain forest mainly vicariantly distributed in two subspecies in NE South America, whereas Norantea goyasensis grows in somewhat higher altitudes on the
Brazilian plateau in habitats with a more pronounced dry season. Outlyers in the distribution of Norantea guianensis were encountered west of the Andes and in
Central America and the Caribbean. Although these fit well in the predicted potential distribution their status
(natural or man-made) remains to be checked .
P0782 – ePoster
West African Plant database – a photoguide and identification tool
Dressler, S 1
Porembsk, S 1
, Schmidt, M 1 , Brunken, U
, Thiombiano , A 1 , Zizka, G 1
1 , Janssen, T 1 ,
1 Senckenberg Research Institute, Botany, Frankfurt/M.,
Germany
The West African Plants Database currently including ca.
7600 photographs of 1280 live plants is an internet photo-database as well as an identification help. The photographs can be accessed via browsing a hierarchical list of taxa and/or searching for specific morphological characters. For the latter the user can choose from a total of 18 easily observable characters regarding flower, fruit, habit, and leaf. The search produces a result page with species names and up to three thumbnail images. By clicking on either of them you get to the species page with all available images and some more data plus links to further internet databases, where you can gain more information on the taxon. The digital photographic documentations are a result of extensive field studies in the course of the BIOTA and SUN projects or were contributed by various colleagues. The photographed taxa are identified by experienced scientists. This website is accessible for anyone interested in the plant diversity of West Africa at
(www.westafricanplants.senckenberg.de). We plan the expansion of the geographical scope to include East and
Central African biomes and welcome further contribution of digital plant photographs with locality data.
P0783 – ePoster
Phylogenetics and biogeography of
Lepechinia
(Lamiaceae); the chloroplast doesn’t cut it
Drew, B 1 , Sytsma, K 1 , Delventhal, N 2
1 University of Wisconsin–Madison, USA;
Manitoba, Canada
2 University of
Lepechinia is a diverse genus in the mint family that consists of about 42 species that range from Northern
California in the Western United States to central
Argentina in South America. Several species of
Lepechinia are valued in the horticultural trade, and
North and South American indigenous groups commonly use Lepechinia for medicinal and antiseptic purposes.
Due to a high morphological diversity and a putative ancient divergence, no synapomorphies for the genus have been found other than a potentially distinctive odor.
Previous morphological (Carl Epling) and cladistic
(Jeffrey Hart) work on the genus left unanswered questions regarding the monophyly of Lepechinia and the intrageneric relationships within the group. Recent molecular phylogenetic studies have shown that
Lepechinia is closely related to Melissa and the large genus Salvia . This study compares phylogenies from plastid (cpDNA; 4 markers), nuclear ribosomal (nrDNA;
2 markers), and low copy nuclear (nDNA; 5 markers) regions. While the nrDNA and nDNA phylogenies are similar, the cpDNA and nuclear DNA regions are shown to be incongruent on multiple levels – from the backbone to the tips. Potential reasons for the differences will be discussed. Based upon evidence from the nuclear and plastid phylogenies, as well as present day distribution, it seems likely that Lepechinia originated in Meso-America
(Mexico) and subsequently dispersed into Western North
America and into South America. The dispersal into
South America probably involved multiple lineages .
Lepechinia is shown to be monophyletic with the inclusion of two monotypic genera, Neoeplingia and
Chaunostoma .
P0784 – ePoster
Systematic studies (morphology, anatomy, palynology and biochemistry) of economically viable grass
Brachiaria mutica
(Forssk.) Stapf in eastern India
Dwari, S 1 , Mondal, AK 1
585

1 Vidyasagar University
Poaceae is the one of the largest family among the
Monocotyledons in the world. So plants of this family are very frequent in the India. Brachiaria mutica , an economically important plant which is considered as one of the best tropical grass for general purposes. The species commonly used as green foliage, grazing and local resources for animal fodder. The present study deals with few systematic parameters viz. morphology, anatomy, palynology, biochemistry of Brachiaria mutica which is commonly known as para grass in Eastern India.
The result shows some distinct morphological and anatomical as well as palynological characters.
Biochemical features show a distinct feature which is different from other monocotyledonous plants. All the details of systematic study of this plant has not been worked earlier.
P0785 – ePoster
New chromosome counts on the genus
Psephellus
(Compositae) in Turkey
Ertugrul, K 1 , Uysal, T 1 , Bozkurt, M 1 , Tugay, O 1 ,
Demýrelma, H 1
1 Selcuk Unýversýty, Science Faculty, Dept of Biology,
Turkey
Chromosome counts were made on somatic metaphases using the squash technique. We report 14 chromosome number records for the genus Psephellus from Turkey.
All of them are first chromosome counts for the studied species. We report the chromosome numbers for 12 species as 2n=30 ( Psephellus taochius, P. turcicus, P. pyrrhoblepharus,P. dealbatus, P. recebi, P. erzincanii, P. schischkini, P. aucherianus, P. hypoleucus, P. straminicephalus, P. holtzi, P. oltensis ), and two species
2n=60( P. eugenii, P. brevifimbriatus ). The prevalence of the basic chromosome number of x = 15, is confirmed.
According to our counts, polyploidy, and especially tetraploidy, occurs in Psephellus and seems to be an important factor for diversification in this genus.
P0786 – ePoster
Geographical speciation in the Mediterranean Basin
(
Reseda sect.
Phyteuma
, Resedaceae)
Martín-Bravo, S 1 , Jiménez-Mejías, P 1 , Escudero, M 2
1
Luceño, M 1
Pablo de Olavide University, Seville, Spain; 2
,
Morton
Arboretum, Lisle, Illinois, USA
Reseda sect. Phyteuma is a natural group composed of 14 species distributed in southern Europe, northern Africa and south-western Asia, with a centre of diversity in the
Mediterranean Basin. With only two widespread species
( R. arabica, R. phyteuma ), most taxa are regional endemics from the Western (mainly Iberian Peninsula and north-western Africa) and Eastern (mainly Turkey and Middle East) Mediterranean. This biogeographical pattern could be tentatively related to two main, disjunct centres of diversification in both sides of the
Mediterranean, as previously reported for other plant
586 groups at different taxonomic levels. Accordingly, both regions constitute hotspots of plant biodiversity within the Mediterranean Basin. This interesting distribution of species diversity and endemism offers an ideal case to study speciation processes in the Mediterranean. We investigated them in a geographical and temporal scenario using a nuclear-plastid molecular phylogeny and analyses of (1) times of diversification based on penalized likelihood and relaxed clock (uncorrelated lognormal) methods, and (2) ancestral range reconstruction, based on dispersal-extinction-cladogenesis and stochastic methods. Reconstruction of ancestral areas inferred a complex pattern of vicariance and long-distance dispersal processes among the two centres of diversity of sect.
Phyteuma in the western and eastern Mediterranean.
Systematic relationships within sect. Phyteuma revealed an additional Mediterranean East–West disjunction at a lower taxonomic level ( R. media – R. orientali s). Low molecular differentiation of R. media populations from the Iberian Peninsula and Macaronesia (Azores and
Madeira) suggests fairly recent long-distance dispersal events to account for the colonization of these oceanic
Archipelagos, as previously described for other
Resedaceae species. Our results, however, are not conclusive to discard an anthropogenic introduction of R. media in Macaronesia, as supported by some authors.
Our time estimates for the origin of the Ibero-North
African endemic R. collina , as well as for R. media and
R. phyteum a, are later than the Mio-Pliocene opening of the Strait of Gibraltar, thus suggesting that long-distance dispersal rather than vicariance may have played a role in shaping the range of these species. Fairly recent geological processes like the Plio-Pleistocene aridification of northern Africa and Middle East could have promoted lineage diversification in sect. Phyteuma in these regions through allopatric divergence of different populations in Mediterranean climate refugia. Likewise, the colonization of south-eastern Europe by R. inodora could be linked with the formation of loess deposits following the drainage of the Pannonian plain in the
Pleistocene. Relationships among the endemic taxa from the Balkans and Anatolia show geographical rather than taxonomical congruence, possibly indicating the influence of geography in the diversification of this species group
P0787 – ePoster
Chromosome evolution in angiosperms and its incidence in lineage diversification rates
Martín-Bravo, S
Fernández-Mazuecos, M
Jiménez-Mejías, P
M 1
1
1
, Escudero, M
4
2 , Mayrose, I 3
, Fiz-Palacios, O
, Valcárcel, V 6
5 , Hipp, A L
, Vargas, P 4
2
,
,
, Luceño,
1 Pablo de Olavide University, Seville, Spain;
Arboretum, Lisle, USA; 3
2 Morton
University of British Columbia,
Vancouver, Canada;
Spain; 5
4 Royal Botanic Garden of Madrid,
University of Uppsala, Sweden; 6 Autonomous
University of Madrid, Spain
Many studies have long argued about the rate of incidence as well as the evolutionary and genomic consequences of polyploidy in flowering plants. As a result, it is nowadays clear that polyploidy is ubiquitous across most, if not all, angiosperm lineages, due to both

paleo- and neopolyploidization events. However, there are still conflicting opinions on whether there is a positive or negative relationship between polyploidization events and diversification rates. Besides this debate, there has been comparatively far less attention on the evolutionary role of other important cytogenetic mutations like single chromosome gains and losses (aneuploidy s.l.) and other types of polyploidy as
‘demipolyploidization’ (half genome duplication). In our study, we try to model the cytogenetic evolution of a wide array of angiosperm groups (15), in order to relate it to their particular evolutionary features, and to draw possible patterns of lineage diversification in relation to different cytogenetic mutations. We span 11 orders of both monocots and dicots at different taxonomic levels ranging from families to sections. In addition, we also included a dataset comprising a representation of all extant angiosperms. We used previous molecular phylogenies and a compilation of chromosome counts for the analyses. We investigated (1) the fitting of eight different models of cytogenetic evolution, (2) the reconstruction of cytogenetic mutation events and ancestral chromosome numbers across the phylogenies
(chronograms with absolute times), and (3) the presence of significant diversification rate shifts and their possible correlations with inferred cytogenetic mutations. Models of chromosome evolution including whole or half genome duplication processes (polyploidization and demipolyploidization) were more frequent than those which did not. In congruence, polyploidization was the most frequent type (in terms of presence / absence) of inferred cytogenetic mutation in the datasets analysed.
Interestingly, the presence of demi-polyploidization, associated with hybridization between different ploidy level and allopolyploidization processes, was inferred in more than half of the datasets. In contrast, the absolute number of chromosome gain and loss events inferred by our analyses was much higher than the absolute number of demi- and polyploidizations, and they seem to be independent of the current chromosome number. These aneuploid processes could be so abundant because they imply far less genomic rearrangement than demi- and polyploidization processes, although they could still promote speciation due to their influence on the chromosome recombination processes. In contrast, the balance hypothesis predicts that additions or substractions of single chromosomes have greater negative impact on the phenotype than whole genome changes, as these aneuploid events are supposed to entail a higher genetic imbalance. Only after the exclusion of certain datasets with especially high aneuploidy rates, genome duplications events become higher than aneuploidy, although not so as expected by the balance hypothesis. Interestingly, while nearly ca. 4000 cytogenetic mutations were inferred for the 15 datasets analysed, only 25 significant or marginally significant shifts in the diversification rate were detected in the nodes of the phylogenetic trees. This result suggests that cytogenetic mutations are usually not sufficient to foster changes in the lineage diversification pattern.
P0788 – ePoster
Los Zuniga Canyon: bryophyte and vascular plant communities in a floristic refuge in the Mexican central highland plateau (State of Queretaro)
Ruiz Herranz, A 1
A 2
, Estebanez Perez, B 1 , Cabrera Luna,
1 Dpto. Biologia, Universidad Autonoma de Madrid,
Madrid, Spain; 2 Herbario de la Universidad Autonoma de Queretaro, Queretaro, Mexico
Although Mexico is considered a megadiverse country, vast extensions in its central plateau are used for agriculture or livestock, and the interpretation of its natural vegetation is problematic. Areas with difficult access play an important role as local biodiversity refuges . Los Zuniga Canyon is about 25 km long and belongs to the San Juan hydrological system (Central
Queretaro). Only some isolated botanical collections, and the inferences on its plant communities shown in the vegetation map of the State, are known. In this study, in addition to compiling herbarium specimens previously collected in the area, we have undertaken seven sampling campaigns in order to collect vascular and bryophyte samples, to assess and describe plant communities through transects (vascular plants) and analysis of subenvironments (bryophytes) . As a result of our new identifications, 76 species (53 vascular plants and 23 bryophytes) are added to the area checklist. Three of them ( Salix aeruginosa, Pogonatum comosum and
Asterella lateralis ) are reported for the first time in the
State. We describe the main bryophyte communities in the Canyon, and provide a more accurate understanding of dominant vascular plant formations, sometimes discordant with those represented in the vegetation map.
For instance, we report the presence of an evergreen oak shrubby community as a result of the anthropogenic disturbance of the evergreen oak forest; a different physiognomy for the xerophilous scrubland, as its lack of
Cactaceae species makes it inadequate to consider it
‘crassicaule’ and we describe a mixed, man-disturbed community, instead of the deciduous tropical forest recorded in the vegetation map . With almost all plants corresponding to the autochthonous neotropical floristic element, and a proportion of nearly 54% plant species restricted to Mexico and neighboring territories, this area is confirmed as a natural refuge of the native flora, and we point out the pressing need for its conservation.
P0789 – ePoster
Phylogenetic analysis of the European and
Mediterranean species of
Erica
L. (Ericaceae) based
1 on non-molecular data
Fagundez, J 1
IE University, Segovia, Spain
The European and Mediterranean species represent a small portion of the vast genus Erica , about 20 species of a total of over 860; however, the different phenotypic aspects of these few species have been thoroughly studied, and they can be used as a representative sample of the overall genus. In this study, a wide set of characters were included in a phylogenetic analysis for the northern species, to test congruence of our analysis with the classical infrageneric classification and with previous analyses of molecular characters and to assess which are the most informative characters with respect to phylogeny of the genus. Seventy characters, covering
587

ecological, phenological, morphological, anatomical, phytochemical, palynological and cytological aspects of the European species of genus Erica , were used.
Phyllodoce caerulea and Daboecia cantabrica were used as the out-group. Fifteen most parsimonious trees were obtained, and the fifty-percent-majority rule consensus tree was calculated. Eight characters were uninformative and sixty percent of the studied characters were highly homoplasious (HI > 0.5). Several clades were supported by the bootstrap analysis but the position of Erica spiculifolia, E. ciliaris, E. umbellata and E. australis was unclear. Some of the clades obtained in our analyses agree with the classical taxonomic groups, but the overall systematic arrangement of the genus should be revised according to these results.
P0790 – ePoster
Study of the relation between public space morphology and biodiversity in the city of Porto
Farinha-Marques, P 1 , Lameiras, JM 1
1 Faculty of Sciences, University of Porto, Portugal
Urban green structures are crucial landscape elements for the promotion and conservation of biodiversity in the ever growing urbanized areas. Landscape architects, architects, urban planners and managers should be able to develop integrated design approaches to promote green structures in the urban realm according to contemporary social needs and grounded on a deep understanding of urban ecosystems. Such approaches should create opportunities for humans, plants and animals, in shared public spaces with the least possible conflict. One way to meet such goals is through a well balanced design of the outdoor space, reflecting the development of a green matrix of multipurpose habitats and social facilities with significant aesthetic quality . Therefore it is important to answer some questions such as: What is the relationship between the spatial character of the urban green structure and its biodiversity? How organization and design of the urban green structure influence the diversity of flora and fauna? Which spatial strategies and tools can be used by planners, designers and managers of the urban green structure to address the question of biodiversity within the city environment? Focusing on the city of Porto, the heart of a metropolitan area of nearly 2 million inhabitants. It is the second largest Portuguese city with a mild Atlantic climate, with several important natural habitats close to each other (river Douro and its mouth, seafront, rocky escarpments) and a variety of man-made habitats with different design character, sizes, locations, and connectivity (public green spaces, relic woodlands, beaches, abandoned farmlands, old gardens, derelict buildings, brownfields and ruderal sites). Its geographic, social and environmental context together with high diversity of situations in a relatively dense urban fabric makes Porto an ideal place for a detailed green structure research in a southern European scenario . Porto green structure (parks, gardens, highway intersections, river corridors, oceanic fronts, cliffs, etc.) was surveyed and classified with particular attention on its design and spatial qualities. Each place assessed according to a set of variables regarding its spatial character, design composition, human use, habitat diversity and species richness. Relationships between biodiversity indicators,
588 ecological factors, and spatial design. Based on the fieldwork and data interpretation, spatial patterns that create opportunities or problems to urban biodiversity were identified, to conclude about which design/ planning strategies have an impact on biodiversity levels .
This research contributes to a better understanding and promotion of the relationship between biodiversity, spatial form, sustainable design and management within the urban realm, particularly for the academic disciplines involved in such areas of knowledge, planners, designers, managers and decision makers.
P0791 – ePoster
A phylogeny of
Solanum
sect.
Crinitum
(Solanaceae)
1 and its relationships within the
Leptostemonum
clade
Farruggia, F 1 , Bohs,
Dept of Biolgy, University of Utah, USA
L 1
Solanum sect. Crinitum is a small section of ca. thirteen species within the enormous genus Solanum . It is composed of small shrubs and large canopy trees found from the Andean cloud forests to the Amazon rainforest and the Brazilian cerrado. The section has received some taxonomic attention but, until recently, has been largely absent from molecular phylogenies. Section Crinitum is most closely related to sect. Androceras within the
Leptostemonum clade, or spiny solanums; however, the relationship between these sections has not been clearly defined until now. Section Crinitum is characterized by its large stature as well as the large flowers and fruits in most species. Here we discuss the species and present data that support the traditional species circumscriptions.
We also provide morphological characters that relate to the principal lineages within the section and discuss the importance of fruit and trichome morphologies. Our data indicate that section Crinitum contains two main centers of diversity, one in the sub-montane humid forests along the eastern slopes of the Andes and another in eastern
Brazil. Restricted species distributions are common in the
Andean species while the lowland species are more widely distributed. These distributions are related to orographic and climatic features of the landscape and shed light into the limited or extensive geographic range of each species. These data help to resolve relationships within the large Leptostemonum clade and will aid in taxonomic revisions of species within Solanum section
Crinitum .
P0792 – ePoster
Revised generic delimitations for
Oxypolis
and
Ptilimnium
(Apiaceae tribe Oenantheae) based on nuclear rDNA ITS and cpDNA sequence data and fruit anatomy
Feist, MA 1 , Downie, S 1 , Magee, A 2 , Liu, M 3
1 Dept of Plant Biology, University of Illinois At Urbana–
Champaign, USA; 2 Compton Herbarium, South African
National Biodiversity Institute, South Africa;
Biology, Harbin Normal University, China
3 Dept of
The Apiaceae are a large economically important family; however, relationships within this family, especially

below the level of subfamily, are poorly understood . The use of molecular data is radically changing the circumscription of subfamilies, tribes, and genera within this family . Fruit anatomical characters have also recently been found to be very useful in delimiting these relationships . Oxypolis and Ptilimnium are two genera within the Oenantheae tribe (subfamily Apioideae) that are endemic to North America . Both are relatively small genera, each with six and seven species, respectively . As is the case with many genera in the Apiaceae, relationships within and between Oxypolis and
Ptilimnium have been unclear . For example, Ptilimnium nodosum and three species of Oxypolis ( O. canbyi, O. filiformis , and O. greenmanii ) share an unusual leaf morphology . Rather than having the pinnately compound leaves that generally characterize the Oenantheae tribe, they have reduced linear, terete, mostly hollow, septate appendages (rachis-leaves) . In a previous study, nuclear rITS data was used to explore relationships within and between these genera and to examine their position within the Oenantheae tribe . A phylogeny based on ITS sequences showed that neither Oxypolis nor Ptilimnium as presently defined are monophyletic . The rachis-leaved
Oxypolis species are more closely related to both the rachis-leaved and compound-leaved Ptilimnium species then they are to the compound-leaved Oxypolis species .
Also, the genera Cynosciadium and Limnosciadium form a strongly supported clade with the rachis-leaved
Oxypolis species and the Ptilimnium species . For this current study, two regions of the chloroplast genome
( trn Q and trn K) were sequenced in order to corroborate and add support to these findings . In addition, fruit anatomical characters were examined . Results of the phylogenetic analysis of the cpDNA data are congruent with results of the ITS data and provide greater resolution within the Ptilimnium / rachis-leaved Oxypolis /
Cynosciadium / Limnosciadium clade . The fruit anatomical characters also provide additional evidence for the non-monophyly of Oxypolis and Ptilimnium. The generic limits of both Oxypolis and Ptilimnium will be changed to reflect these findings.
P0793 – ePoster
Phylogeny and distributed patterns of a cosmopolitan greenbrier family Smilacaceae (Liliales)
Fu, C 1 , Cameron, K 2 , Qi, Z 1 , Li, P 1
1 Institute of Plant Sciences, Zhejiang University, PR
2 China; Dept of Botany, University of Wisconsin,
Madison, USA
Greenbrier (Smilacaceae) currently belonging to the order Liliales, ordinarily including genus Smilax and
Heterosmilax , is a group of worldwide distributed climbing monocot, about 200 species, with alternate leaves characterized by reticulate venation, a pair of petiolar tendrils and usually prickly stems. Although there is a long history of study since Linnaeus named it in 1753, the phylogeny of this dioecious family has never been well resolved. Here, we collected molecular evidence from rDNA (ITS) and chloroplast loci ( mat K, rpl16 intron) of 135 taxa, covered all sections and major distribution zone of the family, Ripogonaceae,
Philesiaceae chosen as outgroup, and reconstructed a sound phylogenetic tree, provided the possible evolutionary history of this family. Our results indicated the phylogenetic relationships, which showed conspicuous geographical pattern within the family: S. aspera with Paleotropical distribution (Mediterranean,
East tropical Africa, India–Himalaya disjunction) is sister to all other taxa of the family. Rest of the family is largely divided into two major clades: Asia and
Americas, with some migration and long distance dispersal events within the two clades. And genus
Heterosmilax formed a monophyletic clade nested within the Asia clade of Smilax . Phenotypic traits according to the phylogeny and more detailed geographical patterns within the clades were found. It is revealed that species in Australia, S. kraussiana in Africa and herbaceous
Smilax in North America migrated/dispersed from Asia.
S. sieboldii and S. scobinicaulis in eastern Asia were found nested in the American clade. S. melastomifolia probably dispersed from South America. For divergence time estimating, maximum likelihood ratio tests rejected a global molecular clock. Therefore, we applied a relaxed molecular clock method to infer the divergence times of all extant lineages of Smilacaceae, calibrating its phylogenetic tree with the fossil record of Ripogonum tasmanicum and the estimated divergence time between
Smilacaceae, Ripogonaceae and Philesiaceae by Janssen and Bremer (2004). Our results suggested that divergence within Smilacaceae started in the late Eocene. Three lineages, the Paleotropical clade, Asia clade and America clade, were formed at the transition time between Eocene and Oligocene, showing a paleotropical distribution of
Smilacaceae before that time. Most of the species diversified in the middle Miocene. A greater richness of species was found in Asia than in Americas. Various phenotypic groups of Smilax , e.g. erect shrub, herb, semiherb, fused perianth ( Heterosmilax ), and panicals were endemic in Asia. Chromosomal polyploidy was also only found in Asia. Finally, a new taxonomic treatment will be given after finishing this project.
P0794 – ePoster
1
A systematic study of
Skimmia japonica
Thunb.
(Rutaceae)
Fukuda, T 1 , Naiki, A 2 , Nagamasu, H 3
Museum Programs and Communications Department,
2
National Museum of Nature and Science, Japan;
Okayama University, Japan;
Museum, Japan
3 Kyoto University
The genus Skimmia Thunb. is an evergreen shrub or tree, occurring in the Sino-Japanese floristic region and its adjacent area. Skimmia japonica Thunb. is its easternmost species, distributed from Sakhalin, Japan and
Taiwan. Such broad distribution range, covering wide climatic ranges and various habitats from coniferous to broadleaved evergreen forest, is a unique characteristic of this species. Skimmia japonica has been divided into 3 varieties in accordance with tree forms and sizes: var. japonica with erect stems, var. intermedia with ascending stems and var.
lutchuensis with erect stems with large size of tree, leaves and flowers. However, because of the lack of distinct characteristics, delimitation of the varieties has not been clear. Molecular phylogenetic analysis of 3 cpDNA regions ( trn Gtrn S, trn Dtrn T, rpl16) in S. japonica revealed 13 haplotypes. Main 5
589

haplotypes (named A to E) had geographic structure.
Haplotype A was distributed from Sakhalin to Taiwan, except part of Pacific Ocean side of Honshu and a part of the Ryukyu Islands. Haplotype B was found along the
Pacific Ocean side from Tohoku to Kii peninsula.
Haplotype C was found from Kyushu to the middle of the Ryukyu Islands, and haplotype D was in the southern half of the Ryukyu Islands. Haplotype E was distributed only in Taiwan, along with haplotype A. Minimum spanning network analysis suggested that haplotype A may be ancestral, and haplotype E, for one hand and haplotypes C, D for other may be derived from haplotype
A. Though haplotype A and B are distributed separately in the eastern Japan, they are found simpatrically in the central Japan. Plants with haplotype A widely occurs from broadleaved evergreen to coniferous forest, and plants of haplotype B grows in deciduous forest, too, not confined to broadleaved evergreen forest. Considering the results of morphological and cytological analysis, plants with haplotype A from Kyushu to the north and plants of haplotype B had same karyotype, and both of the plants had various growth forms, and they were not distinguished by other morphological aspects, either. On the other hand, haplotype C, occurring from northern
Ryukyus to the part of Kyushu, covering subtropical to warm-temperate ranges may compose one common group together with the plants of haplotype D in southern
Ryukyus, together with haplotype A in the northern
Ryukyu Islands (Tokara Isls. and Amami-Ohshima Is.), in having erect stems with large flower size with rather large pollen colporate number. They also had karyotypic variation, differing from the plants of haplotype A from
Kyushu to the north and of haplotype B.
P0795 – ePoster
Brazilian specimens collected by Prince Maximilian of Wied held in the National Herbarium of Victoria
(MEL)
, de Moraes, PLR 2
1
Gallagher, C 1
2
Royal Botanic Gardens Melbourne, Australia;
Universidade Estadual Paulista, Brazil
The National Herbarium of Victoria (MEL) holds approximately 1.2 million specimens. Around 400 000 of these are foreign (extra-Australian) collections, the majority of which derive from the private herbarium of
Otto Wilhelm Sonder (1812–1881). Some 250 000–330
000 of Sonder’s specimens were acquired by MEL in
1883. Once described as ‘the richest of all private botanical collections in existence’, Sonder’s herbarium spans all major plant groups from every reach of the globe, and includes important early 19th-century collections from what were then the least accessible parts of tropical South America, southern Africa and Asia. The significance of the collection is yet to be fully documented; less than 10 per cent of the material has been mounted and databased. As such, the MEL collection has been overlooked in many revisions of major plant groups . MEL holds thousands of important collections from Brazil, including specimens used in
Flora brasiliensis (of particular note is Myrtaceae material studied by O.C. Berg) and specimens from
A.F.M. Glaziou, C.F.P. Martius, A.F. Regnell, L. Riedel,
F. Sellow and J.F. Widgren . A recent discovery at MEL
590 is an important set of Brazilian collections by Prince
Maximilian of Wied (1782–1867). A German explorer, naturalist and ethnologist, Wied was one of the first
European naturalists to visit Brazil. He explored the south-eastern regions of Brazil between 1815 and 1817, returning to Europe with extensive field notes and observations on indigenous South Americans and a wealth of ethnological, zoological and botanical collections. Whilst Wied is relatively well known for his ethnological and zoological studies, his contributions to botany have received less attention. Wied’s botanical specimens are amongst the earliest extant collections from Brazil. These collections are held in several herbaria, mainly BR, GOET, GZU, LE and MEL. At least 1114 collections are known from the literature, encompassing 130 families, 525 genera and 1055 species of vascular plants, and 13 families, 17 genera and 24 species of cryptogams. Over three quarters of these collections have been located, including over 500 nomenclatural types. These collections are a precious record of the flora of the Atlantic rainforest of southeastern Brazil in the early 19th century . Among the least documented of Wied’s collections are those at MEL. A partial search of the MEL collection has revealed over 60 specimens, spanning 22 families, 34 genera and 53 species of vascular plants. Several of these collections have not been located in other herbaria – a tantalising indication of the significance of the Sonder material at
MEL. Other collections are not cited in the literature.
Almost half of the Wied specimens so far located at MEL are nomenclatural types. Ongoing searches at MEL are expected to uncover a further 100 to 150 of Wied’s collections. Details of Wied’s collections at MEL and other herbaria will be published in the Catalogue of
Brazilian plants collected by Prince Maximilian in Brazil.
P0796 – ePoster
Seed micromorphology in
Dactylorhiza
and related genera (Orchidinae, Orchidaceae)
Gamarra, R
P 2
1 , Sanz, E 1 , Esparza, I 1 , Ortuñez, E
1 Universidad Autonoma de Madrid, Spain;
Politecnica de Madrid, Spain
2
1 , Galán,
Universidad
Seed micromorphology of 25 taxa of the genera
Dactylorhiza (incl. Coeloglossum ), Pseudorchis ,
Gymnadenia (incl. Nigritella ) have been examined under light microscopy and SEM . The results show that the sculpturing of the testa cells is a good diagnostic character which allow to recognise these genera. A complete description for each group within the genera, based on qualitative and quantitative characters, is provided. Finally, we compare our results with morphological and molecular data recently published .
The present study aims to survey the seed coat micromorphoplogy using SEM and light microscopy, and provide new insights into its potential taxonomic value.
P0797 – ePoster
Strong genetic structure and phylogeography of
Taxus wallichiana
in Sino-Himalayan region
Gao, L-M 1 , Liu, J 2 , Li, D-Z 3 , Möller, M 4

1 Kunming Institute of Botany,Chinese Academy of
Sciences, Kunming, Yunnan, China; 2 Germplasm Bank of
Wild Species in Southwest China, Kunming Institute of
Botany, CAS, Kunming, Yunnan, China; 3 Graduate
University of the Chinese Academy of Sciences, Beijing,
China; 4 Royal Botanic Garden Edinburgh, Scotland, UK
Abstract: The collision and post-collisional convergence of the Indian subcontinent with Eurasia resulted in the uplift of Qinghai-Tibetan Plateau (QTP) which dramatically changed the topography and climate of
Asia. Climatic oscillations in the Quaternary played a major role in species expansion and population structure.
In this study, the chloroplast DNA trn L-F region sequencing and nine microsatellite loci genotyping of
815 individuals from 43 natural populations of
Himalayan yew ( Taxus wallichiana Zucc.), an endangered tree species occurring in the East Himalayan and Hengduan Mountain (HM) were performed. Twentynine cpDNA haplotypes were detected in the sampled populations, which were divided into two lineages on phylogenetic and network analyses. The distribution pattern of the cpDNA haplotypes showed a strong phylogeographical structure (NST = 0.768, GST = 0.469,
P < 0.01). The divergence time of the two lineages dates back to 3.41–7.61 Ma, supporting the two lineages diverged by the rapid uplift of the QTP. The SSR data of the same samples well correspond with the cpDNA results. Higher level of the genetic diversity was detected in EH populations than that in HM populations. The genetic diversity decreasing westward from China to
Nepal alone the Himalayas was found in EH populations.
Strong natural barrier was found between the two lineages. The mismatch distribution results revealed that the HM lineage may experience a seriously bottleneck effect in the Last Glacial Maximum (LGM), and then a recent rapid range /demographic expansion. And the East
Himalayan lineage probably survived in separate refugia and underwent several expansion / contractions during glacial/inter-glacial cycles in Quaternary.
P0798 – ePoster
Evolution of the basal angiosperm family
Winteraceae on Zealandia
Gemmill, C 1
Demetras, N 1
, Pratt, S 1 , Munzinger, J 2 , Lowry, P 3 ,
1 University of Waikato, New Zealand;
New Caledonia; 3
2 IRD, Noumea,
Missouri Botanical Garden, USA
The oceanic islands of New Zealand and New Caledonia are cut from much the same cloth: they are both off-cast fragments of Gondwana, have endured dynamic and transformative geological processes including periods of submergence, and have been isolated from any other landmasses in the southern Pacific ocean for similar time frames. Furthermore, these islands are intimately connected as they both rest upon the sunken former continent of Zealandia. Evidence of this shared history is reflected by the numerous flowering plant families that are found on both of these sister-Archipelagos. This study focuses on the evolution of the iconic plant family
Winteraceae of Zealandia, and specifically on the sister genera Pseudowintera and Zygogynum (sensu lato).
Winteraceae are an ancient, mostly Southern Hemisphere angiosperm family of evergreen trees, known for their vessel-less wood, peppery scent, remarkable flowers and excellent fossil record. Winteraceae are represented in
New Zealand by a single genus Pseudowintera (horopito) with four extant endemic, well-known species. The case is quite different for the New Caledonian taxa, which reveal a complex taxonomic history. The most recent revisions by Vink were highly radical, placing all members of four previously recognized genera into a single broadly defined genus Zygogynum comprising 18 endemic species, within which he also recognized 13 subspecies and 3 varieties for a total of 30 taxa. It is not clear whether the characters used to delimit these infraspecific entities are constant or if they are phenotypically plastic and hence taxonomically artificial and uninformative. To test Vink’s taxonomic hypotheses and resolve relationships among these taxa, we have sampled Zealandic taxa comprehensively and used both nuclear (ITS) and cpDNA sequence variation.
Preliminary analyses suggest that Zygogynum is not monophyletic with Z. amplecicaule (including all currently recognized infraspecific taxa) sister to
Pseudowintera and the remaining Zygogynum taxa.
Within the large Zygogynum clade, some species are resolved as monophyletic (e.g., Z. comptonii ), while many remain paraphyletic. Overall our analyses suggest that revision of this genus is required. In addition to a revised taxonomic treatment and reconstructed biogeographic and evolutionary scenarios, our work will contribute important new information relevant to the ongoing debate about the extent and duration of the
Oligocene drowning of New Zealand and help address a much-debated question regarding the origin of these taxa: are they continuously inhabiting ancient lineages or is their modern presence the result of more recent longdistance dispersal?
P0799 – ePoster
A morphological study of genus
Colutea
L. (Fabaceae) in Iran
Ghahremaninejad, F
SA 1
1 , Pooyan, P 1 , Assadi, M 2 , Angaji,
1 Tarbiat Moallem University, Iran; 2
Forests & Rangelands, Tehran, Iran
Research Institute of
Genus Colutea contains 28 species, classified in five sections and six subsections. The genus includes ten species in Iran: C. cilicica (sect. Colutea subsect.
Arborescente s), C. persica , and C. gracilis (sect. Colutea subsect. Graciles ), C. buhsei and C. gifana (sect.
Rostrata subsect. Centralasiaticae ), Colutea × variabilis
(sect. Rostrata subsect. Orientales ), C. uniflora , C. komarovii , and C. porphyrogramma (sect. Armata ) and according to recent molecular study C. triphylla (sect.
Oreophysa ). Five of these species (i.e. C. persica, C. gifana, C. uniflora, C. porphyrogramma and C. triphylla ) are endemic for Iran. C. buhsei and C. persica with their vast distributions occupy northern line and S Iran respectively. While C. gifana from NE Iran is an endemic species collected only in a small area. Besides,
C. uniflora turned out to grow in one locality, as well.
However, others involve restricted and narrowed distributions. Nearly all species of the genus are, at first sight, so similar to one another, especially those with the
591

same color flowers. In this study, the herbarium specimens have been analyzed and revised based on 84 morphological features. However, in order to determine sections, subsections and species accurately, 17 of them turned out to be more important. They include spinescence of shoots, termination of the keel top, pubescence of ovary, shape and length of wings, length of flowers, color of flowers, number of flowers in inflorescence, length of calyx teeth and kind of incisions between them, number of leaflet pairs, size and shape of leaflets, pubescence of leaflets, arrangement of leaves, way of bark peeling, size of fruits, length of fruit stipe, termination of fruit apex, and color of calyx hairs.
Additionally, the Latin and English diagnoses of taxa, respectively from 'Flora Iranica' (Rechinger 1984) and from 'The Genus Colutea L. a Monograph' (Browicz
1963) have been considered. Therefore a key to determine species growing in Iran has been made. To define the position of the localities of all species in the country, which were read from the sheets' labels have been plotted on maps. Since recent molecular study proved Oreophysa to be synonymized with Colutea (sect.
Oreophysa ) triphylla and as it is similar to Colutea in some features, it was also observed in this investigation as a member of the genus, yet considered it as distinct species in this genus.
P0800 – ePoster
Australasian Loganiaceae: molecular phylogenetics and biogeography of Loganieae
Gibbons, KL 1, 2 , Henwood, MJ 1 , Conn, BJ 2
1 University of Sydney, Australia; 2 National Herbarium of
NSW, Sydney, Australia
Loganiaceae is a predominantly tropical to subtropical family with occasional extensions into higher latitudes .
Areas of highest generic diversity include the Americas,
Africa and Australasia . The morphological heterogeneity of the family has long been noted and recent molecular and morphological cladistic analyses have confirmed the polyphyletic nature of Loganiaceae as traditionally circumscribed . A monophyletic core Loganiaceae has been resolved, and is best represented in Australia and
New Zealand by Logania R.Br., Mitrasacme Labill. and
Geniostoma J.R.Forst. & G.Forst
. Tribal circumscriptions have varied widely . As a result of recent molecular phylogenetic analyses, the circumscription of
Loganieae has been broadened to include all Australian and New Zealand genera except the pantropical
Strychnos L. (Strychneae) . Loganieae, therefore, provide an excellent model with which to explore the biogeographical history of the Australasian region .
Generic limits and relationships, however, remain largely untested. Nuclear ribosomal and chloroplast nucleotide sequence data were used to conduct maximum parsimony and Bayesian phylogenetic analyses of Loganieae to test generic limits and to resolve intergeneric relationships .
Logania Section Logania and Logania Section
Stomandra (R.Br.) DC. each appear to be monophyletic .
However, the relationship between the two sections of
Logania remains uncertain . The Hawaiian endemic genus
Labordia Gaudich. was found to nest within the more geographically widespread Geniostoma , as previously proposed on morphological data . The south Western
592
Australian endemic Mitreola minima B.J.Conn was placed not with Mitreola L. but sister to Mitrasacme,
Schizacme Dunlop and Phyllangium Dunlop . Historical biogeographic patterns and the evolution of selected morphological characters within Loganieae will be presented.
P0802 – ePoster
Taxonomy, phylogeny and biogeography of
Leiothrix
(Eriocaulaceae)
Giulietti Harley, AM 1,3 , Andrade, MJG 1 , Queiroz, LP 1
1
Scatena, VL 2 , Berg, CVD 1
,
Universidade Estadual de Feira de Santana (UEFS),
Brazil;
Brazil; 3
2 Universidade Estadual Paulista (UNESP),
Royal Botanic Gardens, Kew, UK
The genus Leiothrix was first described by Ruhland in
1903 with 28 species, of which eight were first published in the same work and the remainder were species removed from Paepalanthus . It was characterized by having pistillate flowers with free petals as in
Paepalanthus , but with a gynoecium in which the styles are fused into a column whose branches become free at different levels. The author recognized five subgenera:
Calycocephalus (= Leiothrix ), Rheocaulon , Eleuthandra ,
Stephanophyllum and Psilanthus . Following the Index
Nominum Genericorum , the genus contains 72 validly published species and 44 infraspecific taxa. During the last 20 years, the species of Leiothrix have been the subject of diverse studies, including morphology, anatomy, chemotaxonomy, taxonomy and, more recently, morphological and molecular cladistics. These studies, associated with the molecular data presented in this paper, will constitute the basis for a monograph of the genus to be published in Flora Neotropica . Leiothrix is a well supported monophyletic group, both in morphological and molecular analyses. It falls within
Eriocaulaceae subfam. Paepalanthoideae and is distinguished from Syngonanthus and Comanthera by the possession of pistillate flowers with petals fused in the median region. It also differs from both these genera and from Paepalanthus by the form and arrangement of the styles, and by the flattened stamen filaments and basifixed anthers and by the striate seed coat. The taxonomic was based on intense fieldwork in the area of distribution of Leiothrix , and all the collected material is deposited in the herbaria SPF and HUEFS. The study was complemented by an analysis of specimens of the genus deposited in 51 herbaria in Europe and the
Americas. In this work, 38 species of Leiothrix are recognized, and a further eight infraspecific taxa, as a result of the discovery of one new species and five new combinations, the synonymization of 24 species and 15 infraspecific taxa. Also, ten species and one infraspecific taxon were transferred to other genera, including L. arechavaletae the only species of L . subgen. Psilanthus , which has been transferred to Syngonanthus , while seven taxa are considered dubious. Diagnostic keys, descriptions, notes and illustrations, as well as distribution maps, are all presented. All the species of
Leiothrix occur in Brazil, and only L. flavescens (Bong.)
Ruhland also occurs outside, in Venezuela, Guyana and
Peru. The major concentration of species is found in the
Espinhaço mountain chain, particularly in the Serra do

Espinhaço in Minas Gerais State (27) and in the Chapada
Diamantina in Bahia State (7). The molecular phylogeny of Leiothrix was realized using 23 species, representative of four subgenera. The analyses based on a mixture of individual and combined sequence data using plastidial
DNA ( trn Ltrn F) and nuclear (ITS), employing maximum parsimony and Bayesian inference. The monophyly of Leiothrix was confirmed, but among subgenera, only L. subgen. Stephanophyllum emerged as monophyletic. Above all, the genus showed in all analyses, a dichotomy, formed of two clearly delimited and well-supported clades, defined by their geographical distribution and associated to microendemism in the
Espinhaço mountain chain.
P0803 – ePoster
Preliminary studies reveal new species within southern African
Pachycarpus
E. Mey. (Apocynaceae:
Asclepiadoideae)
, Nicholas, A 1 , Lamb, J
1
Glen, M 1 1 , Shuttleworth, A
University of KwaZulu-Natal, South Africa
1
Recent specimens of Pachycarpus collected at Highflats
(South Africa, KwaZulu-Natal) suggest that these populations may be allied to, but distinct from, P. asperifolius and P. scabe r. In order to test this hypothesis and at the same time propose a rational classification for the Highflats populations it was decided to do an extensive morphological investigation complimented by and integrated with gene sequence studies of the nuclear ribosomal ITS region. Molecular data remains equivocal on the exact evolutionary relationships and distinctness of the Highflats populations; however, morphological studies confirm the unique status of this taxon and suggest that it should be described as a new species, viz. acidostelma . In the process of this investigation, it was observed that there were some major morphological differences between the northern and southern populations of P. asperifolius . It was found that, although the molecular data were largely unresolved, there was a
0.37% genetic difference between northern and southern populations of P. asperifoliu s. Based on this, and the morphological differences, it was decided that the northern populations of P. asperifolius be recognised as a distinct species, viz. validus , validating older classificatory hypotheses of Schlechter and N.E. Brown.
P0804 – ePoster
Divergence timing of the several north Americaneastern Asian disjunct lineages within
Aristolochia subgenus
Isotrema
(Aristolochiaceae)
Gonzalez, F 1 , Wagner, S
Samain, MS
Wanke, S 2
3 , Isnard, S
2 , Symmank, L 2 , Salomo, K 2
2 , Rowe, N 3 , Neinhuis, C 4
,
,
1 Universidad Nacional de Colombia, Colombia; 2 Institut für Botanik, Technische Universität Dresden, Germany;
3 Dept of Biology, Plant Seed Research, Ghent University,
Ghent, Belgium; 4 CNRS, UMR AMAP, University
Montpellier, Montpellier, France
With c. 69 species from temperate and tropical Asia, and
North and Central America, Isotrema is the second largest subgenus of Aristolochia (Aristolochiaceae). The monophyly of Isotrema has received strong support from morphological and molecular data; however, the specieslevel relationships and the divergence timing within this subgenus have not yet been addressed . We performed parsimony- and Bayesian phylogenetic analyses of 31 species (17 from the New World) based on three chloroplast regions ( mat K gene, the trn K intron, and the trn K-psbA spacer), and estimated the times of divergence within the subgenus using BEAST and r8s . The analyses indicate that the Asian and the New World species are intermingled; thus, they do not conform two geographically exclusive clades. The first split within the subgenus led to the differentiation of the North American
A. reticulata + A. serpentaria clade, which occurred during the Oligocene, c. 30.2 mya . The remaining species confirm a North American–Eastern Asian (NA–EAS) subclade and a Mexican/Central American (MCA, exclusively Neotropical) subclade,which diverged during the Early Miocene, c. 21.8 mya . Within the NA–
EAS subclade, the main divergence event occurred during the mid-Miocene, c. 16.8 mya; two Asian–North
American disjunct lineages are nested within the latter subclade: (1) a western North American–Eastern Asian pair formed by A. californica as sister to most of the
Asian species, a split occurred by the mid-Miocene, c.
13.2 mya; and (2) A. manshuriensis , from Eastern Asia and A. tomentosa , its sister species from eastern North
America, a split that occurred by the Late Miocene, c. 6.7 mya. The striking morphological similarity between A. manshuriensi s and the eastern North American A. macrophylla is not in agreement with the molecular data, and could be the result of morphological stasis in these two species. Within the MCA subclade, A. panamensis , the southernmost species of the subgenus, differentiated from the remaining Neotropical species during the mid-
Miocene, c. 13.4 mya; disjunction events between species from Asia and the MCA clade were not detected.
Most of the latest speciation events in all major lineages from the subgenus have occurred during the
Plio/Pleistocene. Interestingly, although the number of species in Asia (c. 49) is much higher than that in the
New World (c. 20), most of the diversity in growth form
(including lianas and self-supporting treelets and shrubs), leaf architecture (including pinnately veined blades), inflorescence position (including cauliflory), and perianth elaboration (including tails and fungus-like, club-like or blister-like processes on the floral fauces) has only occurred within the MCA clade.
P0805 – ePoster
Burmeistera
(Campanulaceae) revisited: a taxonomic
1 account based on the Colombian species
Gonzalez, F 1 , Garzon, J 1
Facultad de Ciencias, Instituto de Ciencias Naturales,
Universidad Nacional de Colombia, Colombia
After the cosmopolitan Lobelia L. (c. 405 species), and the Neotropical Siphocampylus Pohl (c. 231 species) and
Centropogon C. Presl. (c. 212 spp.), Burmeistera (c. 100 spp.) is the fourth largest genus of the subfamily
Lobelioideae (Campanulaceae) . The genus was erected
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by the Colombian botanist J. J. Triana in 1854, based on
B. ibaguensis . Molecular and morphological characters indicate that Burmeistera is monophyletic, unlike the related genera Centropogon, Lobelia , and
Syphocampylus , which have shown to be polyphyletic.
The only infrageneric classification of Burmeistera , based on the presence or absence of a tuft of long hairs on the ventral anthers, is in conflict with molecular data .
The species of Burmeistera are herbs, subshrubs or sometimes shrubs up to 5 m tall, with whitish to yellow
(sometimes red) bat-pollinated flowers. Most of the species have restricted distribution in low to high elevation (200 to 3000 m) cloud forests in Honduras,
Costa Rica, Panama, Colombia, Venezuela, Peru and
Ecuador . We revisited the taxonomy of the genus based on the study of the 47 Colombian species (c. 50% of the genus), thirty of which are exclusive to this country .
Most of the Colombian species (c. 80%) are restricted to the Western and the Central Cordilleras, which could be related to a preference for volcanic soils . We described six new species, proposed twelve new synonyms, and clarified the nomenclatural and taxonomic issues related to the type species of the genus . Furthermore, we designated lectotypes for B. ceratocarpa Zahlbr., B. ibaguensis , and B. rivina E. Wimm., as well as a neotype for B. mutisiana (Kunth) E. Wimm. A careful examination of the hair tuft in the ventral anthers during floral development shows that this diagnostic character needs to be studied in preanthesis, as these hairs shrivel during anthesis . The presence of this hair tuft in the ventral anthers could be plesiomorphic in Burmeistera , as it is also present in the related genera Centropogon,
Lobelia , and Syphocampylus. We propose that the ebracteate peduncle is a potentially phylogenetically informative character within Burmeistera .
P0806 – Poster
Flora of the State Park El Faro and its potential use
Gonzalez-Ibarra, M
Hernandez, A 1
1 , Silva-Torres, B 2 , Chimal-
1 Universidad Autonoma Metropolitana-Xochimilco,
2 Mexico; Universidad Autonoma Metropolitana-
Iztapalapa, Mexico
The State Park of ‘El Faro’, located in Tlalmanalco, State of Mexico, is part of the Ecological Reserve of the Sierra
Nevada, which consists of different biological corridors, product of the principal reason of the enormous biological richness of our country: the meeting of two kingdoms biogeograficos (the Neártica and the
Neotropical). Nevertheless, in spite of possessing legal protection for the conservation of these corridors there has been observed in the last years an intensive growth of the human settlements that they find around it. This demographic growth attracts undoubtedly direct consequences in the ecosystem of the reserve principally in loss of biodiversity. The Sierra Nevada headquarter of
‘El Faro’ and of the historical volcanoes Popocatépetl and Iztaccíhuatl, it constitutes a very important area for the capital of Mexico for the mass of oxygen that it provides to it and for constituting one of the most important reserves of water, since it provides to great part of the east of the metropolitan zone. Nowadays faced with the imminent deforestation that we face for diverse
594 social factors such as poverty, lack of employment, immoderate felling etc., it is necessary to have information about the condition of the flora of the state park ‘El Faro’ and to contribute to the analysis of the local vegetation, as well as to help to the inventory of its biodiversity. Periodic samplings were made in the park with the intention of contrasting and monitoring the variety of existing species. Likewise there decided the potential use of flora of the park. The assignment as
Natural Protected Area, of ‘El Faro’, has as principal aim to protect and preserve the natural environment, in order to support the balance and the continuity of the ecological processes, and of its elements, by means of the implementation of protection programs, managing and environmental education and scientific research; to assure the use and sustainable utilization of the resources for the future generations.
P0808 – ePoster
1
Genetic diversity and phylogeography of wild-sown and cultivated coconuts (
Cocos nucifera
L.)
Gunn, B 1 , Crisp, M 1 , Baudouin, L 2 , Olsen, K 3
Research School of Biology, Australian National
University, Canberra, Australia;
France; 3
2 CIRAD, Montpellier,
Dept. of Biology, Washington University, St
Louis, MO, USA
The coconut ( Cocos nucifera L.) is a pantropical strand plant, colonizing sandy insular beaches in the humid tropics. Cocos is a monotypic genus in the Cocoseae tribe (18/ca. 200 spp.) within the Arecaceae family .
Phylogenetic studies support its sister relationship to
Syagrus , a Neotropical genus, sharing a common ancestor about 35 MYBP, though the crown group age of
Cocos is about 11 MYBP . Fossil evidence indicates that members of the Cocos lineage were present in South
America, India, New Zealand and Australia . Coconuts are adapted to drift-dispersal by ocean currents; however, human activities both historically and today have also aided its spread and impacted its phenotypic and genetic structure . Coconuts are traditionally classified as 'Talls' or 'Dwarfs' based on tree habit . Morphological examination reveal two predominant fruit types, attributed to Polynesian terminology: niu kafa are characterized by their elongated, triangular fruits with large proportion of fibrous husk and niu vai, whose fruits are rounded with large proportion of liquid endosperm .
The niu kafa form is interpreted as the naturally evolved coconut, under natural selection for dissemination by sea currents whilst the niu vai form evolved from domestication under human selection for greater volume of delicious coconut water . Here we investigate the genetic diversity of coconuts, the impact of domestication, introgression and the taxonomic implications for this species. We used polymorphic microsatellite markers on 1322 coconut samples representing phenotypic and genetic variation worldwide to examine the geographical location of the center(s) of domestication and its progenitors . Bayesian analyses of population structure revealed two major subpopulations corresponding to the Pacific and Indo–Atlantic oceanic regions . Haplotype networks based on chloroplast and nuclear markers are used as a complementary dataset to examine the coconut's phylogeography.

P0809 – ePoster
Population structure of the A-genome group in
Glycine
subg.
Glycine
(Fabaceae)
Gunner, S 1 , Straub, S 2 , Doyle, J 1
1 Cornell University, USA; 2 Oregon State University,
USA
Sister to the cultivated soybean, Glycine subg. Glycine contains several diploid genome groups of species native to Australia and Papua New Guinea. Each genome group is defined by its species being interfertile and has been validated as monophyletic with molecular study. Many of these groups contain species contributing to the
Glycine polyploid complex. The A-genome is the largest genome group and consists of about seven species. G. syndetika, G. canescens , and G. clandestina are the three
A-genome representatives that contribute to the polyploid complex. Due in part to Glycine subg. Glycine being predominantly autogamous with common cleistogamy, population structure of the A-genome species has been poorly understood. Such structure is important for elucidating the formation of the entire polyploid complex. To uncover population structure we analyzed variation at thirteen nuclear microsatellite markers across
97 accessions. The variation supports multiple subdivisions underlying the A-genome species.
Furthermore, this study initiates exploration of the relationships among the A-genome species and the interconnected members of the Glycine polyploid complex.
P0810 – ePoster
A synopsis of Chinese
Macromitrium
(Bryopsida,
Orthotrichaceae)
Guo, S 1 , Cao, T 1 , Lou, Y 1
1 College of Life and Environmental Sciences, Shanghai
Normal University, China
Sixty taxa (including species and varieties) of
Macromitirum have been reported from China. Among them, twenty-nine taxa were originally described as new based on specimens collected from China. According to our revisional work of Chinese Macromitrium , we found a new species Macromitrium hainanese Guo & He, fifteen new synonyms, four new records to China, made one new combination, and we considered 27 species and five varieties to be taxonomically recognizable in China.
In the present paper, we also listed these recognizable species, and gave a key to them.
P0811 – ePoster
Incongruence between gene trees and species tree suggest rapid diploid and polyploid speciation in the
N Hemisphere
Achillea millefolium
complex
(Asteraceae)
Guo, Y-P 1 , Zhang, Y-F 1 , Wang, S-Z 1
Ehrendorfer,
, Vogl, C
F
2 ,
3
1 College of Life Sciences, Beijing Normal University,
Beijing, China;
Genetics, University of Veterinary Medicine, Vienna,
Austria; 3
2 Institute of Animal Breeding and
Faculty Centre of Biodiversity, Faculty of Life
Sciences, University of Vienna, Vienna, Austria
The N Hemisphere Achillea millefolium aggregate exhibits complex morphological and genetic variation and broad ecological amplitude. To understand evolutionary history of such a species complex, we here study its diploid and polyploid species and populations within a phylogeographic and population genetic framework. We analyzed haplotype sequence data of two nuclear, the ncpGS and the SBP genes, and three plastid fragments, trn H-psbA, trn C-ycf6 and rpL16 in 30 populations of seven diploid and 11 tetra- and hexaploid taxa/cytotypes throughout the distributional range of A. millefolium agg. To better estimate species tree from gene trees, three congeneric diploid species outside A. millefolium agg. were also analyzed with the nuclear gene sequences. In the diploids, gene trees of the two nuclear and the plastid loci are incompatible with each other and with the species tree inferred from the previous
AFLP analyses. Incomplete lineage sorting and intragenic recombination (found in the ncpGS gene) are inferred on the basis of gene tree incongruence. The present sequence data are thus less suited to recover the relationships of diploid species in A. millefolium agg. than the previous AFLP data. An analysis using a probabilistic model for population demographic history indicates a large ancestral effective population size and short intervals between speciation events. Such a scenario explains the incomplete lineage sorting we observe. The combined data, the nuclear and plastid haplotypes and the AFLPs, suggest that many of the polyploid taxa in A. millefolium agg. have hybrid origins.
Little lineage sorting indicates rapid diploid species differentiation in A. millefolium agg. The observed patterns of sequence variation among closely related species may have been caused by the climate fluctuations during and after the Quaternary ice ages. For non-model organisms, marker polymorphisms from multiple unlinked loci, e.g., AFLP markers, combined with haplotype sequence data from some nuclear and plastid loci seem useful and cost and time efficient. We thus advocate such a combination to determine phylogenetic relationships among recently radiating species as well as to understand the demographic history.
P0814 – ePoster
Phylogenetic relationships within Sarcolaenaceae
(Malvales), Madagascar’s largest endemic plant family, based on molecular sequences data
Haevermans, T
Applequist, W 2
Phillipson, PB
, Aubriot, X 1
1,2
1 , Lowry II, PP 1
, Cruaud, C
, Haevermans, A 1
, Schatz, GE
3 , Hong-Wa, C 2
, Rouhan, G 1
2 ,
,
1 Muséum National d’Histoire Naturelle, Département
3
2
Systématique et Evolution, CNRS/MNHN Origine,
Structure et Evolution de la Biodiversité, Paris, France;
Missouri Botanical Garden, St Louis, MO, USA;
Genoscope, Centre National de Sequençage, Evry,
France
595

Preliminary results from a molecular phylogenetic reconstruction using a broad sampling of Sarcolaenaceae
(Malvales), Madagascar’s largest endemic family, reveal intricate relationships and provide new insight into longstanding questions regarding generic delimitations. The major clades based on molecular evidence are largely congruent with morphological groups and provide strong support for the generic circumscriptions proposed in a recent series of taxonomic revisions. All genera form moderately to well supported monophyletic groups, with the exception of Rhodolaena , which appears to be biphyletic (although support for this hypothesis is weak).
The earliest diverging taxa lineages comprise genera that are represented by only a few species, with the exception of Schizolaena , the largest member of the family. All the taxa possessing a complete involucre, a structure that envelopes the mature fruit and a distinctive feature of the family, form a very well supported clade. Taxa assigned to Leptolaena s.l. are shown to be paraphyletic with respect to Sarcolaena , a finding that has necessitated generic recircumscription and led to the resurrection of the genera Mediusella and Xerochlamys .
P0815 – ePoster
Evaluating the feasibility of using candidate DNA barcodes in discriminating species of the genus
Dioscorea
Hang, Y 1 , Sun, X 1 , Guo, J 1 , Peng, B 1 , Bai, M 1
1 Institute of Botany, Jiangsu Province and Chinese
Academy of Sciences, China
DNA barcoding is an aid to taxonomic identification by using a short, standard DNA region. Here, the applicability of three DNA regions ( rbc L, mat K, and psb Atrn H) as the barcodes was tested for identifying species within the genus Dioscorea . Based on assessments of the success rates of PCR amplifications, the sequence quality, extent of specific genetic divergence, DNA barcoding gap and ability for species discrimination, mat K is recommened for barcoding
Dioscorea . 149 plant samples of Dioscorea including 38 species, 7 varieties, 1 subspecies were collected from the main distributing areas in China. mat K successfully identified 93.3% of all samples, comparable to 68.5% of rbc L and 34.9% of psb Atrn H. The combination of two sequences or three sequences would achieve higher success rates of species discrimination, all with 100% success rates of species discrimination except rbc L and psb Atrn H. But the cost of experiments would be much higher using two or three sequence combination than single sequence. Therefore, our research indicated that the mat K is a powerful, though not perfect, barcode for
Dioscorea identification by comprehensively considering the factors of ability for species discrimination and the cost of experiments.
P0816 – ePoster
A new classification of
Hyptis
and allied genera
(Lamiaceae) based on molecular phylogenetic and morphological data
Harley, RM den Berg, C 2
1 , Pastore, JFB 2 , Forest, F 1 , Paton, A 1 , van
596
1 Royal Botanic Gardens, Kew, UK; 2 Universidade
Estadual de Feira de Santana, Brazil
The family Lamiaceae occurs almost worldwide, comprising seven subfamilies, about 240 genera and over
7000 species. It has recently undergone major changes of delimitation, and further changes at a lower level are continuing. The present account is a molecular phylogenetic study of one subtribe, Hyptidinae, belonging to the tropical tribe Ocimeae, subfamily
Nepetoideae. The Hyptidinae is a neotropical group of about 400 species, currently divided into nine genera, occurring from the southern United States and the
Caribbean to Peru and Argentina, with a few species extending, mostly as weeds, into the Old World. The major centre of diversity of the genus is in Brazil. The largest genus is Hyptis Jacq., currently with over 280 species, which display much variation in habit, vegetative and reproductive morphology and habitat preferences, while almost all species share a characteristic corolla form which supports an explosive pollination system. Within Hyptis , 24 sections are currently recognized and their inter-relationships and relationships with other genera have long remained uncertain. As the subtribe apparently diverged early, compared with other groups of Ocimeae, knowledge of its relationships could shed light on the origins of the tribe as a whole. The analysis of the ITS dataset sampled, overall, 180 species of Hyptidinae, representing all nine genera and 22 of the 24 sections of Hyptis . This analysis was used to select a smaller sample of 69 species for further analysis, using plastid ( trn L-F, mat K, trn S-G, trn D-T) and nuclear (ETS and ITS) markers, which demonstrated the subtribe Hyptidinae to be well sustained as monophyletic. Most of the smaller genera were also found to be monophyletic after Hypenia vitifolia (Pohl ex Benth.) Harley, Eriope simplex (A. St-
Hil.) Harley and Hyptis eximia Epling had been reassigned to their correct genera. The genus Hyptis , however, was shown to be paraphyletic, forming at least nine separate lineages, while the small genus Peltodon , although monophyletic, was nested within the clade which included the type species of Hyptis . The recognition of most of these lineages as new genera presents no great difficulty as seven of them are composed of one or more groups currently treated as sections or subsections of Hyptis , while two small genera are composed of species forming new groupings. Hyptis sect. Polydesmia Benth. and Hyptis sect. Mesosphaeria
Benth. were both found not to be monophyletic and the species will be redistributed among two newly recognized genera: Polydesmia and Mesosphaerum . As a result, subtribe Hyptidinae is augmented to 19 genera.
Hyptis sensu stricto will now consist of about 9 sections and almost 150 species, including Peltodon reduced to sectional status.
of those species which are pantropical weeds, two: Hyptis suaveolens (L.) Poit. and H. pectinata
(L.) Poit. will become species of Mesosphaerum P.
Browne, Hyptis spicigera Lam. will be transferred to the new genus Polydesmia , while Hyptis capitata Jacq., H. lanceolata Poir. and H. brevipes Poit. will remain unchanged. A generic key to Hyptidinae and distribution maps will be presented.

P0817 – ePoster
An ISSR investigation of north American species of grapeferns (
Sceptridium
: Ophioglossaceae)
Hauk, W 1 , Cao, D 1
1 Denison University, USA
In North America the center of Sceptridium species diversity is in the eastern half of the continent. Unlike
Botrychium s.s., species concepts in Sceptridium are poorly investigated, perhaps due to high amounts of intraspecific morphological variation and a paucity of species-specific characters. We investigated six diploid
North American Sceptridium species ( S. dissectum, S. oneidense, S. biternatum, S. multifidum, S. rugulosum, and S. lunarioides ) using inter simple sequence repeats
(ISSRs) to test species concepts. Geographically representative samples from each species were amplified using a total of 9 ISSR primers. A mean of 28.8 loci per primer was generated, and nearly 100% of loci were polymorphic. Although preliminary NJ and MP analyses revealed most accessions of the same species clustered together, few fixed species-specific markers were detected. These Sceptridium species may represent taxa that have not yet completed primary divergence, and this may help explain the difficulty of distinguishing species morphologically.
P0818 – ePoster
Spore output and arrangement patterns of the elaters of selected liverworts
He, Q 1 , Zhu, R-L 1
1 East China Normal University, China
The liverworts are one of three phyla in the bryophytes, and may be the earliest diverging lineage of land plants.
Their reproduction is mainly by means of spores, which are usually released by elaters in the capsules. The spore output and elater arrangement patterns are important in comparative studies on spore dispersal, fecundity, population maintenance and classification system, but they are still poorly known. The spore output of 50 selected liverworts including 37 leafy taxa and 13 thallose ones from China were investigated by artificial counting and haemacytometer counts in 10% sucrose solution. The mean spore output per capsule ranges from
222 in Cyathodium cavernarum and Cyathodium smaragdinum to 465000 in Calypogeia tosana .
Compared with other leafy families investigated,
Lejeunea ceae has a much lower and more stable spore output, ranging from 257 in Cololejeunea magnilobula to
5038 in Ptychanthus striatus . Except for Ptychanthus striatus the remaining 25 Lejeunea ceous species investigated have a mean value of less than 3000. Out of
50 species investigated, Calypogeia tosana, Pallavicinia lyellii and Bazzania tridens show higher mean spore output over 100000. Among 11 genera of thallose liverworts, Cyathodium and Ricciocarpus produce larger spores, but have a lower mean spore output ranging from
222 to 592. This paper also reports morphological diversity and arrangement patterns of the elaters of selected liverworts. Some new arrangement patterns of elaters in Lejeunea ceous taxa, including Trocholejeunea sandvicensis with 42 elaters per capsule in a '1+3+5' or
'2+4+6' pattern, Mastigolejeunea indica with 54 elaters per capsule in a '1+3+5+6¡' or '2+4+6' pattern, and
Cheilolejeunea gaoi with 26 elaters per capsule in a
'1+2+2+2' or '2+2+2' pattern, are described and illustrated for the first time. Photographs of spores and elaters under SEM and LM are also provided.
P0819 – ePoster
Phylogenetic relationships of Taxaceae and
Cephalotaxaceae based on leaf and wood characters
Ghimire, B 1 , Lee, C 2 , Heo, K 1
1 Kangwon National University, Chuncheon, Republic of
Korea; 2 Korea National Arboretum, Pocheon, Republic of Korea
Except from the major split into the six genera, phylogenetic relationships within genera of Taxaceae and
Cephalotaxaceae have remained enigmatic for the most part. The aim of this study is to understand the morphological and anatomical differences through the leaf anatomy, stomata morphology, wood anatomy, and vascular pattern of seed in genera of Taxaceae and
Cephalotaxaceae. It also contributes some useful information for the phylogeny of these two conifer families. Four species of Taxus : T. baccata, T. caespitosa, T. cuspidata including three varieties, var. caespitosa , var. latifolia , var. nana and T.
wallichiana ,
Torreya nucifera , Amentotaxus yunnanensis and
Cephalotaxus harringtonia including one variety nana were considered for the study. Leaf anatomy was performed by microtome section, stomata structure was observed by scanning electron microscope and wood anatomy was carried out by sliding microtome sectioning method. We found differences among the studied taxa in epidermal layer, resin duct, structure of stomata, and vascular pattern of seed. The development of stomata in rows and differentiated by papillose accessory cells is the most distinguish feature of Taxus compared to other three genera. Resin duct is totally absent in Taxus but it was well-developed in Amentotaxus , Torreya and
Cephalotaxus . Arrangement of stomata and structure of subsidiary cells of Torreya is very unique than other genera in which stomata pore is deeply sunken and protected by loosely arranged filament like special band of cells. Some common wood characters are revealed: absence of resin duct, clearly developed growth rings, helical thickening in tracheary wall and uniseriated ray
(only Torreya bears both uniseriate and biseriate ray).
Taxus and Torreya have pure helical thickening in tracheid wall whereas Cephalotaxus bears combination of both annular and helical thickenings. Amentotaxus seems to have annular thickening in tracheid wall. The pure helical type tracheid wall thickening in Taxus and
Torreya may be preceded from annular type of
Amentotaxus via mixed type in Cephalotaxus.
P0820 – ePoster
Systematics and morphology of selected angiosperm reproductive structures from the Eocene Laguna del
Hunco flora, Chubut Province, Argentina
597

Hermsen, EJ 1
Johnson, KR 4
, Gandolfo, MA 1 , Wilf, P 2 , Cúneo, NR 3 ,
1 Cornell University, USA;
3
2 Pennsylvania State
University, USA; Museo Paleontológico Egidio
Feruglio-CONICET, Argentina; 4 Denver Museum of
Nature and Science, USA
The early Eocene (ca. 51.91 Ma) Laguna del Hunco (LH) flora of Chubut Province, Argentina, is preserved in tuffaceous sediments and was deposited in a caldera-lake environment along with an assemblage of insects and some vertebrates. The paleoflora is highly diverse, with taxa that today can be characterized as belonging to three different groups of rainforest plants displaying different climatic tolerances and geographical distributions: 1) subtropical to tropical Australasian taxa; 2) neotropical taxa; 3) and Gondwanan temperate taxa. Despite the excellent preservation, diversity, and interesting biogeographic connections of the flora, very little research has been done documenting the abundant angiosperm reproductive material that has been collected from LH. E.W. Berry named and illustrated the taxon
Schmidelia eduliforma based on a single infructescence in 1925 (he transferred the species to the extant genus
Allophylus (Sapindaceae) in 1938); he also described four other taxa, three species of Carpolithus and one species of Malvacarpus , in each case based on one to several specimens of angiosperm fruits or seeds. Only one of these species – the fruit taxon – has since been formally revisited; it has been revised several times, and was most recently included in the extant genus Orites
(Proteaceae). The only angiosperm reproductive material that has been newly described includes staminate inflorescences and ovulate ‘cones’ assigned to the extant genus Gymnostoma (Casuarinaceae). Work on a large suite of leaves, infructescences, capsules, buds, and a flower of Eucalyptus (Myrtaceae) is ongoing. Given that reproductive organs are, in general, the most taxonomically diagnostic and the most phylogenetically informative structures in the flowering plants, it is critical that the LH angiosperm reproductive structures be fully documented so that this information can be incorporated into a more complete understanding of the composition and biogeographic relationships of the LH paleoflora. In this poster, we will introduce selected undescribed angiosperm reproductive material from LH that is currently under investigation, and revisit one previously described taxon. Special attention will be given to inflorescences and infructescences bearing bicarpellate capsules and thought to have affinities to the extant genus Weinmannia or other Cunoniaceae; the aforementioned ‘Allophylus’ infructescences, which bear multicarpellate fruits; and several types of winged fruits.
The latter include specimens with what is interpreted as a robust, persistent calyx reminiscent of the fruits of
Ceratopetalum (Cunoniaceae), and examples of another type of fruit with a tri-lobed wing and planar grouping of four seeds that resembles the arrangement of seeds in fruits of some extinct Juglandaceae from the Northern
Hemisphere, although its affinities require further investigation.
Of these reproductive structures, the fruits with likely affinities to Ceratopetalum show a clear
Australasian rainforest signal. Where the other taxa fit into the emerging concept of the LH flora remains less clear, and will require more precise phylogenetic placement to decipher.
598
P0821 – ePoster
Climatic and biodiverstiy changes during the
1
Holocene in southern Chile: a leaf remains analysis
Hinojosa, LF 1 , Prez, F 2
Universidad de Chile, Instituo de Ecologia y
Biodiversidad, Chile; 2 Pontificia Universidad Católica de Chile, Institutp de Ecologia y Biodiversidad, Chile
Climatic change is considered one of the major drivers of unprecedented biodiversity changes, being probably the main focus of discussion in science and both political and social spheres. A key issues for predicts the impact of climatic change on biodiversity is understand how the distribution and/or ecological requirements of species could be affected. An interesting approach to give answers to the future interaction between climate and biota distribution come from analogous conditions in the past, examining the effect that they had in the composition, structure and richness of the biota. Under this context, the western forest region in Chile represents an ideal scenario to study the paleobiogeographic history of their communities. Presentday rain forests of southern
South America are separated from other forested regions in the continent by more than 1,000 km of arid and semiarid lands. This vicariant distribution is due to the development of hyperarid climates in the central Andes during the Pliocene–Pleistocene transition, resulting from the strong rain shadow produced by the Andean uplift.
Because of this geographic isolation and the buffering effect of the Pacific Ocean on the climate of the western margin of the continent; mid latitudes (35˚42’S) in the
Chilean Coastal Range have been postulated as an important biotic refuge during the Pleistocene glaciations. The main goal of this work was to reconstruct the climate and vegetation history in south and central Chile, by the analysis of leaf fossil remains in
Río Caunahue outcrop (40˚S). Paleoclimatic and paleodiversity changes was performed by co-existence and rarefaction analysis. Río Caunahue outcrop corresponds to approximately 13 m of laminated silts and clays with ash layers and organic horizons with an age going from 13110 to 4460 14C years BP. The main leaf remain recovery from Río Caunahue correspond to
NothoFagus dombeyi , Amomyrtus luma, Aextoxicon puntctatum, Eucryphia cordifoli a. Biodiversty increase significantly, associate with a warming condition.
Acknowledgments: Fondecyt 1090339; Instituto Milenio de Ecología y Biodiversidad (IMEB P0- 5002) & PFB-23
(Conicyt)
P0822 – ePoster
Phylogenetics and biogeography of
Ludwigia
1
(Onagraceae)
Hoch, PC 1 , Barber, J 2 , Diazgranados, M
Missouri Botanical Garden, USA; 2
2
St Louis University,
USA
Ludwigia (Onagraceae) is a diverse and ancient genus comprising 82 species in 23 sections. Fourteen of the sections are monotypic, and only three have more than five species. Nine sections are endemic to or centered in
South/Central America, three in North America, four in

Africa, three in Asia, two in the Paleotropics, and two are cosmopolitan. We present a preliminary partial phylogeny that will eventually enable us to test the validity of this highly fragmented classification, determine relationships among the sections and species, address the role of polyploidy and hybridization in the evolution and radiation of the genus, determine which area harbors the most plesiomorphic section(s) and may represent an ancestral area, and how and when the genus radiated from there to other areas. Based on preliminary data, Ludwigia is strongly supported as sister to the remainder of family Onagraceae, and is thought to have diverged from the rest of the family 80–93 MYA, probably in South America. Recent monographs are available for some sections, but the genus as a whole has never been revised thoroughly, and the only synopsis of the genus was published almost 50 years ago. New molecular sequence data are combined with abundant comparative data from anatomy, palynology, embryology, and cytology, to begin to construct an evolutionary scenario for Ludwigia , to re-examine its classification and biogeography, and possibly to provide valuable information concerning control measures for several invasive species of Ludwigia worldwide.
P0823 – ePoster
Dispersal has shaped the distribution patterns in
Ranunculeae DC. and in the cosmopolitan genus
Ranunculus
L.
Emadzade, K 1 , Hoerandl, E 1
1 University of Vienna, Vienna, Austria
Today it is widely accepted that disjunct distributions can be explained either by fragmentation of widespread ancestors by vicariant (isolating) events or by dispersal across a barrier. Molecular-based phylogenetic studies based on DNA sequences and estimates of divergence times of lineages supported the role of dispersal as a primary process shaping distribution patterns in both animals and plants. These studies provide evidence supporting a hypothesis of transoceanic dispersal versus vicariance. Ranunculeae DC. comprise 19 genera and about 650 species. Ranunculus is the only genus distributed on all continents. In contrast, most other genera in this tribe have very restricted distributions, and the monotypic genera are endemic to small areas. The distribution patterns of this tribe provide a model system for studying vicariance versus dispersal. We combine here the results from molecular dating and biogeographical analyses to provide a comprehensive hypothesis of the history of Ranunculeae. To estimate divergence dates, Bayesian uncorrelated rates analyses and four calibration points derived from geological, fossil and external molecular information were applied.
Parsimony-based methods for dispersal–vicariance analysis (DIVA and Mesquite) and a maximum likelihood-based method (Lagrange) were used for reconstructing ancestral areas. Our study suggests a
Northern Hemisphere origin for the Ranunculeae in the
Eocene and a weakly supported vicariance event between
North America and Eurasia. The Eurasian clade diversified between the early Oligocene and the late
Miocene, with at least three independent migrations to the Southern Hemisphere. The North American clade diversified in the Miocene and dispersed later to Eurasia,
South America and Africa. Results of biogeographical analyses of Ranunculus combined with molecular dating suggest multiple colonizations of all continents and disjunctions between the northern and the southern hemisphere. Dispersals between continents must have occurred via migration over land bridges, or via transoceanic long-distance dispersal, which is also inferred from island endemism. The cosmopolitan distribution of Ranunculus is caused by transoceanic and intracontinental dispersal, followed by regional adaptive radiations.
P0824 – ePoster
Apomixis, autopolyploidy and maintenance of genetic diversity cause a geographical parthenogenesis pattern in
Ranunculus kuepferi
(Ranunculaceae)
2 , Ladinig, U 2 , Hoerandl, E 1
1
Cosendai, AC 1 , Wagner, J
University of Vienna, Dept of Systematic and
Evolutionary Botany, Austria; 2 University of Innsbruck,
Institute of Botany, Austria
Geographical parthenogenesis (GP) is a phenomenon that sexual and asexual populations of the same species complex do not share the same distribution area. In general, asexuals occupy higher altitudes and latitudes and are more widespread. The alpine species Ranunculus kuepferi is a good model for studying GP, since polyploid asexual populations are widespread throughout the Alps, while diploid sexuals are confined to small refugial areas. Following Baker’s law which allows the establishment of a new population with a single seed, apomixis could provide an advantage in colonization and might explain why the apomicts are more successful.
Genetic diversity might be relevant for surviving genetic bottlenecks and occupation of different ecological niches.
We investigated the mode of reproduction via Flow
Cytometry (FC), Flow Cytometric Seed Screening
(FCSS) and experimental tests of self-compatibility, the origin of polyploids and genetic diversity of populations was analyzed with molecular methods AFLPs (dominant marker) and five microsatellite loci (SSRs). These studies were conducted on 60 populations out of the whole distribution range of the species. We assessed facultative apomixis in tetraploids as a majority of the seeds are produced via gametophytic apomixis with either pseudogamous or autonomous endosperm formation, but also partial apomixis was observed. The combination of apomixis and self-compatibility allows for uniparental reproduction. Facultative apomixis allows for flexibility in the reproductive system and helps to maintain genetic diversity in founder situations. Autopolyploidy was brought into evidence by the lack of private SSR alleles and AFLP fragments in the polyploids. Genetic diversity and population genetic structure suggest that genetic divergence within the species complex is very poor and that tetraploids have been formed multiple times.
Apomictic populations have distinct local gene pools, probably as a result of multiple founder events, but they show almost no clones and a genetic diversity similar to that of sexuals. It is the combination of autopolyploidy, facultative apomixis, and maintenance of genetic diversity that assure the success of apomictic cytotypes.
599

P0825 – ePoster
Searching for the roots of the Arctic flora: biogeographic connections to the Himalayas
Hou, Y 1 , Mirre, V 1 , Sletten Bjoraa, C 1 , Guo,
Y 2
, Popp, M 1
, Brochmann, C 1
1 National Centre for Biosystematics, Natural History
Museum, University of Oslo, Oslo, Norway; 2 College of
Life Science, Beijing Normal University, Beijing, China
Two main hypotheses have been suggested for the origin of the arctic flora: (i) in situ persistence/adaptation of a local Tertiary forest element, consisting of species which occurred in open patches in the forest, and/or (ii) immigration from southern alpine regions, consisting of species preadapted to cold and open habitats. Hulten hypothesized that many contemporary arctic plant species descend from alpine ancestors, which occurred further south in Asia and migrated northwards to
Beringia in the late Tertiary or early Quaternary, from where they radiated east- and westwards and reached circumpolar distributions before the onset of the
Pleistocene glaciations. To test these hypotheses we will conduct a comparative phylogenetic and phylogeographic analysis of several plant groups comprising both arctic species and species occurring further south in Asia, with particular emphasis on the
Hengduan Mountains in the Himalayas (e.g. the genera
Cassiope, Diapensia , and Braya ). The Hengduan
Mountains, which are situated in the southeast Qinghai-
Tibetan Plateau, constitute the major part of the so-called south-central biodiversity hotspot in the Himalayas. This region has been inferred as an important refugium throughout the Pleistocene climatic cycles, as well as a centre of plant diversification during the uplift of the
Qinghai-Tibetan Plateau and the Pleistocene interglacials. It may therefore have served a source for many arctic plant species. By using state-of-the-art molecular phylogenetic/phylogeographic methods and dating tools, we will (i) investigate the connection and the direction of migration between the Hengduan
Mountains and the Arctic (which not necessarily was from the south to the north), and (ii) estimate the timing of migrations to distinguish between the hypotheses of in situ survival of old Tertiary elements in the Arctic versus late Tertiary/early Quaternary (or even later) origin of the arctic lineages. We currently search for appropriate lowcopy nuclear DNA genes, such as RNA polymerase genes. Field sampling in the Hengduan Mountains is carried out in 2010 and 2011.
P0826 – ePoster
Systematics analysis on some species of Cycadaceae by ISSR markers
Huang, Y-Y 1 , Mo, P-Q 2
1 Zhongkai University of Agriculture and Engineering,
China; 2 Guangxi Polytechnic College of Agriculture,
China; *author lqxhyy@yahoo.com.cn for correspondence:
Cycadaceae are antique seed plants with high value for viewing and science. At present, there is hot disputations
600 about its taxonomic problem. In this paper, ISSR markers was used to study taxonomy and relationships of some species of Cycadaceae. The results were as follows: 11
ISSR primers, which were chosen from 100, were found to produce cleanly, repeatably and stably polymorphic segments, and they produced 115 of 132 amplified fragments, the polymorphic ratio reached 87.1%. It explained the genetic distinction and relationships of 29 species of Cycadaceae effectively. The similarity coefficients of 29 species ranged from 0.40–0.83, which indicated the materials have obvious species difference.
Based on the international general standard: similarity coefficient below 0.67 is species level, 0.67–0.9 is considered to be subspecies, variety and form level, similarity coefficient above 0.91 is the different individual genetic difference of subspecies, variety and form. According to three plant taxonomic grades, subspecies, variety and form can be classified particularly as follows that similarity coefficient is the scale 0.67–0.75 is subspecies genetic difference level, similarity coefficient 0.76–0.82 is variety genetic difference level and similarity coefficient 0.83–0.9 is form genetic difference level. On the basis of above standard we establish each cycad classification positon:
C. xilingersis and C. longiconifera are subspecies of C. segmentifida ; C. segmentifida is original subspecies; C. longata is variety of C. segmentifida ; C.segmentifida
is typical variety; C. guangdongensis, C. taiwaniana and C. szechuanensis are independent species; C. hainanensis is variety of C. taiwaniana and C. taiwaniana is original variety; C. longipetiolula is independent species; C multifrondis is form of C. longipetiolula ; C. longipetiolula is original form; C. multipinnat a is variety of C. micholitzii ; C. micholitzii is original variety; C. brevipinnata and C. ferruginea are subspecies of C. miquleii ; C. miquelii is original subspecies; C. parvula and C. balansae are independent species. C. pavulus is independent species, C. shiwandashannia is the variety of C. balansae and C. diannanensis is the subspecies of
C. balansae . Some cycad species which were thought have obvious difference in phenotypic character and have not any disputation in past time, but their similarity coefficients are high – up to the level of subspecies and variety – analysed from this situation, these disputed species have no difference to those cycad species which were thought that have clearned classification positon in genetic level, to these no disputed species in past, we have given some advices for their classification position, so that to learn the relationship of every species of
Cycadaceae well and drawup protective measures effectively .
P0827 – ePoster
Terminalia procera
Roxb. – a distinct species than
1
Terminalia catappa
L.
Ingle, PA 1 , Dhabe, AS 1
Dept of Botany, Dr Babasaheb Ambedkar Marathwada
University, Aurangabad (MS), India
Terminalia catappa L. ( Terminalia badamia Tul.) and
Terminalia procera Roxb. ( Terminalia copelandii Elmer) are the well known species of genus Terminalia (family
Combretaceae). Terminalia catappa is popularly known as Deshi Baadaam and T. procera as Safed Bombay.

Kernels of both the species are edible and rich source of proteins and fats. Gamble (1992) placed Terminalia catappa as a synonym of Terminalia procera , Gupta and
Singh (2005) have shown the anatomical similarities between these two species and suggested to merge these species. Present paper deals with detailed morphological, anatomical (transverse section of bark, petiole and leaf; transverse section, transverse longitudinal section and radial longitudinal section of wood and maceration of bark and wood) and dermatological studies of Terminalia catappa and Terminalia procera . The structures were compared in sections and the different types of cells were compared in maceration. Phytochemical analysis of these species were restricted to phenolics especially phenolic acids, glycoflavones and aglycons. It was observed that, these species differ morphologically – as to size and shape of leaves, colour of lower surfaces of leaves; size and shape of the fruits; anatomically – as trichomes, size and shape of vessels, ray parenchyma; phytochemically – phenolic acids, glycoflavones and aglycones . It was concluded that, these two species are distinct.
P0828 – ePoster
Floral and vegetative development in
Ceratophyllum demersum
(Ceratophyllaceae) with special reference to the phylogenetic position
Iwamoto, A 1 , Izumidate, R 1 , Otani, A 1
1 Tokyo Gakugei University, Japan
, Katase, C 1
Ceratophyllum has been placed in a key position within the phylogeny of angiosperms based on the results of molecular analysis. Recent analyses reported a sister group relationship of Ceratophyllum to the eudicots.
There are other possibilities, in particular, a sister group relationship to the monocots is probable. Therefore, the morphological features of Ceratophyllum are important for elucidating the phylogenic position of the genus. We have already observed the floral development of
Ceratophyllum (Iwamoto et al. 2003), but some of them remain to be observed, especially anatomical observations are necessary. In addition, it is necessary to reevaluate its morphological features in phylogenetic context. In this study, we observed in detail the developmental anatomy of lateral organs initiated at nodes of Ceratophyllum demersum , including vegetative buds, staminate and pistillate flowers. The primordium of vegetative buds was initiated at every node with a subtending leaf, while floral primordia were not found at every node and lacked a subtending leaf. The vegetative buds were often produced singly at each node, but all flowers were accompanied by vegetative buds at the same node. These observations imply that the vegetative buds are derived from main axillary buds, and their accessory buds develop into the flowers. The staminate flowers of C. demersum are composed of more than 10 stamens and are surrounded by more than 10 tepals
(sometimes regarded as bracts). The pistillate flowers are composed of one pistil and are surrounded by more than
10 tepals. The staminate floral apex developed bract primordia first, then stamen primordia. In the early development stages, the tepal and stamen primordia were initiated on the abaxial side of the floral apex and only later on the adaxial side (unidirectional). In the pistillate floral apex, the tepal primordia also showed unidirectional initiation. The unidirectional development in staminate and pistillate flower of Ceratophyllum are in common with the flowers of Acorus , the most basal monocot, and some species of Piperales, a group considered to be close to monocots, which may imply a closer relationship between Ceratophyllum and monocots than with other possible sister groups. We will compare the floral development of Ceratophyllum with those of
Acorus and Alismatales, which is placed as the second most basal group in monocots, and Ranunclales, the most basal eudicot, and discuss the relationship among them in phylogenetic context.
P0829 – ePoster
Molecular phylogenetic and morphological studies on
Trentepohliaceae with special reference to the central pit structure of
Phycopeltis
and its allies
, Shinji, H 2 , Miho, N-T 3 , Hiromi, T 4
1
Izumi, S 1
Dept of Biological Science, Graduate School of Science,
Hiroshima University, Japan; 2 and Health Association, Japan;
Hiroshima Environment
Fellow of Natural History Museum and Institute, Chiba,
Japan; 4
3 Cooperative Research
Miyajima Natural Botanical Garden, Graduate
School of Science, Hiroshima University
The family Trentepohliaceae (Ulvophyceae,
Chlorophyta) consists of ca. 70 species within six genera containing many common aerial algae, and most of the species are widely distributed, especially in tropical and subtropical regions of the world . Although 14 species of five genera have been previously recorded in Japan, only two of Trentepohlia forming visible colonies with macroscopic filaments have been reported . The results of our recent field investigations indicate the presence of unreported species of Trentepohlia and Phycopeltis. In the present study, the morphological features of these species were investigated using field specimens and cultured clones with TEM observation, and their phylogenetic position based on 18S rDNA sequence data was also resolved. The phylogenetic tree suggested two distinct clades: (1) one consists of Phycopeltis and some species of Trentepohlia , (2) the other Printzina ,
Cephaleuros and the other species of Trentepohlia as previously reported by López-Bautista et al. (2006) and
Rindi et al. (2009) . Based on the TEM observation for several strains, two types of interconnection by plasmodesmata were recognized: (1) a distinct central pit which is a structure of a group of plasmodesmata in central portion of the transverse walls between vegetative cells, and thinner than the outer border and (2) a group of plasmodesmata with no small central pit . These results suggest that the former clade would be characterized by not only a pedicellate zoosporangium opening by terminal pore (apical pore) but also occurrence of a distinct central pit, which contains an inner ring of wall material which encircles the central pit . Ultrastructural observations also indicate that the central pit of members of the former clade is unlike the pit connection (or pit plug) of red algae, because of the absence of core and cap .
601

P0831 – ePoster
A phylogenetic study of subgenus
Polypompholyx
: a parallel radiation of
Utricularia
(Lentibulariaceae) throughout Australasia
Jobson, R 1
1 National Herbarium of NSW, Sydney, Australia
Phylogenetic relationships among the 37 recognised taxa of Utricularia subgenus Polypompholyx were assessed by cladistic analysis of DNA sequences from plastid rps 16 and trn L-F. We found strong jackknife support for a monophyletic subgenus Polypompholyx lineage. Within the section Pleiochasia , we found two supported clades, generally differing in a more northern or southern distribution. Despite high levels of morphological heterogeneity and convergence, we found some cladespecific character homogeneity within these two clades, particularly that of growth and bladder-trap form, and floral structure. Bladder-trap form corresponds most strongly with terrestrial v. aquatic habits. The evolution of filiform corolla appendages corresponds with floral colour, and is possibly associated with sexual mimicry, with those of the upper corolla arising twice independently.
P0832 – ePoster
How much similarity is needed to be a good food mimic?
Jersakova, J 1 , Jürgens, A 2 , Šmilauer, P 1 , Johnson, SD 2
1 University of South Bohemia, Faculty of Science, Ceske
Budejovice, Czech Republic; 2 University of KwaZulu
Natal, Pietermaritzburg, South Africa
Most orchids with deceptive pollination mechanisms exploit the food-foraging behavior of pollinators. Some orchids are Batesian floral mimics that attract pollinators by imitating flowers used as sources of food for pollinators. It has been shown that pollinators are often unable to distinguish between these orchid mimics and their rewarding models. In our presentation, we evaluate visual and olfactory cues in Batesian mimicry systems to answer the question: how much similarity is needed to be a successful mimic? We conducted a unique in situ behavioural experiment to test the role of colour and shape matching between the pink flowers of Disa pulchra (an orchid mimic) and Watsonia lepida (an irid model). We offered plastic model flowers of various spectral and shape properties to long-tongued flies.
Transparency and lack of UV reflectance of plastic models turned out to be an essential prerequisite to induce pollinator interest. Models with colours indistinguishable from those of Watsonia model in a fly vision model were intensively visited and probed by flies, which tolerated relatively large variation in overall brightness and saturation. Presence of nectar guides and floral shape improved the attractiveness of model flowers to flies. Our results demonstrate that matching of the colour spectra of an orchid’s flowers to those of a rewarding species is sufficient for the evolution of a
Batesian mimicry system.
602
P0833 – ePoster
Phylogenetic studies in
Tamarix
: a molecular approach to Mediterranean species
Villar, JL 1 , Gaskin, JL 2
Martínez-Flores, F 1
, Juan, A
, Crespo, MB 1
1 , Vargas, MA 1 ,
1 University of Alicante, Spain;
USA
2 USDA-ARS-NPARL,
The taxonomy of the genus Tamarix is considered rather complex by different authors (cf. Bunge 1852; Baum
1978). About 200 species have been described in the last two centuries, but this number could be dramatically increased after considering the infraspecific taxa
(subspecies, varieties and forms). Nevertheless, many of these taxa have been lately synonymized according to the priority principle. Nowadays, different authors recognized between 60 and 100 species in this genus.
Some taxonomical groups are still conformed by quite similar taxa, which are weakly separated by morphological characters, mainly based on floral characters. However, the high variability of these floral characters, even within a single sample, makes very difficult the separation between close related species (e.g.
T. gallica – T. canariensis – T. arborea / T. ramosissima – T. chinensis – T. hohenackeri / T. africana – T. tetragyna / T. passerinoides – T. macrocarpa – T. aucherana , among others). Moreover, the recently proved possibility of hybridation (Gaskin & Schaal 2002, Gaskin & Shafroth
2005) adds another point of difficulty to Tamarix taxonomy. Systematic, evolutionary and biogeographic hypotheses for Tamarix have not been tested in a molecular phylogenetic framework to date. Molecular phylogenetic analysis could play a key role for a better understanding of this genus. Here we present the first phylogenetic hypothesis of Tamarix with special emphasis in the Western Mediterranean species, based on plastid ( trn S-G, psbDtrn T, rpl32-trnL, trn Q-rps16) and nuclear (ITS) markers. Systematic implications regarding the naturalness of sections based on our results are discussed. This work has been conducted with the resources provided by the research project ‘Taxonomy, systematics and phylogeography of the Mediterranean
Tamarix species’ (CGL2008-05056) .
P0834 – ePoster
Phylogenetic relationships of American
Sarcocornia
: an approach of morphological character evolution
1
Guilló, A
JC 1
1 , Juan, A 1
, Crespo, MB 1
, Alonso, MA 1
University of Alicante, Spain; 2
, Lledó, MD 2 , Agulló,
Royal Botanic Gardens,
Kew, UK
The taxonomy of the genus Sarcocornia A.J.Scott is considered as complex by different authors, especially for the American taxa. In fact, it does not exist a general consensus about the American taxonomical identity and, therefore, their proper nomenclature. About eighteen different taxa have been named to date in this continent.
Conversely, only two species are widely accepted in
European continent: S. perennis (Mill.) A.J.Scott and S. fruticosa (L.) A.J.Scott, which could be distinguished

according to vegetative and reproductive features.
Additionally, these two European taxa have been cited for several American floras. Moreover, several subspecies and varieties have been also described on the basis of small morphological differences which are mostly due to ecological causes. Some morphological characters based on vegetative, floral and reproductive features (e.g. habit, leaf apex, width and length inflorescence or size and indumentum seeds) were also analyzed to discriminate among all the studied taxa.
Phylogenetic relationships among American and
European taxa have been assessed using several nuclear and plastid molecular markers. Our first results pointed out the existence of two monophyletic clades according to their geographical origin: America and Europe.
Moreover, biogeographical implications are also discussed within American Sarcocornia group. Finally,
Parsimony and Bayesian analyses were performed. The molecular data were compared with a matrix of basic morphological characters.
P0835 – ePoster
Asymmetrical hybridization between monocarpic mass-flowering shrub and polycarpic perennial herb in
Strobilanthes
(Acanthaceae)
Kakishima, S 1 , Murata, J 1
1 Botanical Gardens, University of Tokyo, Japan
The evolution of life histories is very important for plants that cannot move. The life history of each species has evolved to adapt to habitats. Therefore, it is predicted that a difference of life histories may effect a reproductive isolation between two species, which are closely related and grow sympatrically. Studying natural hybridization between species with different life histories can reveal the role of life histories as reproductive isolation. Several species of Strobilanthes (Acanthaceae) have a plietesial life history – a life history in which almost all individuals simultaneously flower, set seed and die once every three to sixteen years. Because a parallel evolution of this life history was suggested by molecular phylogeny of Strobilanthes , this genus is the best material to study evolution of the life history. In this study, we took up S. flexicaulis and S. tashiroi .
Strobilanthes flexicaulis has a plietesial life history, flowering once every six years, whereas S. tashiroi is a polycarpic perennial herb. These species are very similar but distinguishable by pollen and bract morphology, and grow sympatrically on Okinawa Island, Japan. In sympatric populations, putative hybrids between S. flexicaulis and S. tashiroi were recognized by intermediate pollen and bract morphology. To reveal the origin of putative hybrids, we performed morphometric analysis, cpDNA and nDNA sequencing, cleaved amplified polymorphic sequence (CAPS) analysis, and field observations. Principal component analysis on 18 morphological characters, as well as molecular hybrid index based on 11 nDNA locus calculated by HINDEX
(Buerkle 2005), revealed that most of the putative hybrids were either halfway between putative parents or relatively close to S. flexicaulis morphologically and genetically. Furthermore, an assignment test calculated by NewHybrids (Anderson and Thompson 2002) showed that the putative hybrids were F1, F2, backcross to S. flexicaulis , or later generation hybrid. Not only all individuals of F1 and F2 hybrids but also most of the backcross to S. flexicaulis have S. tashiroi haplotypes.
This result shows that hybridization occurs regularly by pollination from S. flexicaulis to S. tashiroi and hybrids.
Based on pollinator observations in the field, honeybees and moths were frequently found to visit flowers in mass-flowering years but rarely observed in other years.
This may cause unidirectional hybridization by pollinators carrying massive amounts of pollen from S. flexicaulis to S. tashiroi and hybrids in mass-flowering years, during which the flowers of S. flexicaulis greatly outnumber S. tashiroi and hybrids. In other years, when flowering is limited to S. tashiroi and hybrids, the expected backcross to S. tashiroi does not occur. Because of low outcrossing rates in non-mass-flowering years, backcrossing would take place only in mass-flowering years. These results suggest that the difference of life histories cause asymmetrical hybridization between S. flexicaulis and S. tashiroi .
P0838 – ePoster
Taxonomic studies on the liverwort
Trichocolea
Dumort. (Marchantiophyta: Trichocoleaceae) in
Japan
Katagiri, T 1 , Miyauchi, H 1 , Deguchi, H 1
1 Graduate School of Biol. Sci., Hiroshima University,
Japan
Three species of the genus Trichocolea Dumort.: T. tomentella (Ehrh.) Dumort., T. pluma (Reinw., Blume &
Nees) Mont., and a new species ( Trichocolea sp.) are shown to occur in Japan. Previously, only one widely distributed species, Trichocolea tomentella , was known from Japan (Horikawa 1934) . Recently Kikukawa (2007, graduation thesis; unpublished) reported a second species, T. pluma , from the southern part of Japan, primarily distinguished by its distinctly dilated septa of the leaf cilia . Inoue (1978) distinguished T. pluma from
T. tomentella by its dilated septa of cilia of leaves, although Grolle (1977) and Pippo (1984) treated them as
T. pluma -complex or T. tomentella , s. lat . Furuki &
Higuchi (1996) noted the presence of a single eye-spot on the oil bodies in T. pluma , while no such eye-spot was reported on the oil bodies in T. tomentella (Müller 1939) .
Based on fresh material collected in Japan and examination of the type specimens, we reached the conclusion that the salient morphological differences between T. pluma and T. tomentella are sufficient to distinguish them at the species level. In addition, during our revision of the genus Trichocolea in Southeast Asia, we carried out field research in 2009 and found a new species of Trichocolea from Mt. Kaikoma, Southern
Japanese Alps. Trichocolea pluma (Reinw., Blume &
Nees) Mont. is distinct from the other species of the genus Trichocolea in Japan in having a single eye-spot on the oil bodies and thick-walled cells with dilated septa of the leaf cilia . The Japanese specimens previously reported as T. tomentella by Hattori (1951), who noted a single eye-spot on the oil bodies, are misidentified T. pluma . The species is rather common on Honshu,
Sikoku, Kyushu, and Ryukyu Isls. in Japan. Trichocolea tomentella (Ehrh.) Dumort. has leaf cilia with thin-walled cells and the septa are not dilated; oil bodies without an
603

eye-spot; the cuticle of the cilia weakly striolateverrucose; and epidermal cells of the capsule wall without thickenings . This species is also widely distributed and more common in the Northern part of
Japan. The new species ( Trichocolea sp.) is similar to T. tomentella in appearance by its regular branching habit, but distinctive in the massive cuticular verrucae on the cilia, and linear to semiannular thickenings in the epidermal cells of the capsule wall . It has been found on wet boulders on steep mountain slopes, under the vegetation of Abies mariesii, Tsuga diversifolia , and deciduous broad leaved trees as represented by Acer spp .
This species is distributed only in the Southern Japanese
Alps, central Honshu, Japan.
P0839 – ePoster
The evolution of unusual shoot organogenesis in
Podostemaceae
Katayama, N 1 , Koi, S 2 , Kato, M 3
1 Graduate School of Natural Science and Technology,
2 Kanazawa University, Kanazawa, Japan; Graduate
School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, Japan; 3 Dept of Botany,
National Museum of Nature and Science, Tsukuba, Japan
Podostemaceae is a family of unusual aquatic eudicots called river weeds. The plants grow submerged on rocks in rapids and waterfalls, mostly in tropical and subtropical regions. They have unusual determinate shoots, which are morphologically diverse among the subfamilies. The subfamily Tristichoideae (including
Terniopsis ) and Weddellinoideae have typical shoot apical meristems (SAMs) that produce laminar leaves on their flank. On the other hand, in Podostemoideae
(including Hydrobryum and Cladopus ), the shoots lack recognizable SAMs, where new leaves arise below the base of older leaves, and cease to grow after forming several filiform leaves. To infer the evolution of novel shoot organogenesis of Podostemaceae, we investigated the mechanisms underlying shoot development by expression analysis of of key developmental regulatory gene orthologs in model eudicots: SHOOT
MERISTEMLESS (STM) and WUSCHEL (WUS), which are involved in meristem initiation and maintenance, and ASYMMETRIC LEAVES1/ROUGH
SHEATH2/PHANTASTICA (ARP), which promotes leaf identity. We isolated homologs of STM, WUS, and
ARP from Terniopsis minor of Tristichoideae, and
Hydrobryum japonicum and Cladopus doianus of
Podstemoideae and performed phylogenetic analyses to confirm orthology. Expression patterns were examined with in situ hybridization and RT-PCR. In the SAMmediated shoots of T. minor , like in model plants, STM and WUS orthologs were expressed in the SAM. In the
SAM-less shoots of H. japonicum and C. doianus , STM and WUS orthologs were expressed in the initiating leaf and bract primordia. As they developed, WUS expression disappeared and STM expression was restricted to the basal part of the primordia, whereas ARP was expressed in the distal part in a complementary pattern to STM expression. The results suggest that the leaves and bracts of the SAM-less species produce cryptic meristems near their bases, which bulge as a new shoot apex and subsequently differentiate into apical leaf or bract. The
604 leaf/shoot mixed organs repeatedly form via cryptic meristems at their base, resulting in a ‘sympodiallybranched’ SAM-less shoot. We propose the evolutionary model of novel shoot organogenesis in Podostemaceae that the repetitive leaf-like shoot formation of
Podostemoideae is derived from the sympodial shoot branching of Tristichoideae and Weddellinoideae. By contrast, the flower of the SAM-less species, which develops as normally as in model plants, the expression patterns of the three genes in the floral organs and floral meristem were consistent to the pattern in other angiosperms. The leaves of the SAM-less shoots in
Podostemoideae have a mixture of leaf and shoot identity. We suggest that these organs evolved by the transformation of the shoot apex into a single, terminal leaf-like organ by losing STM/WUS expression and gaining ARP expression in the organ. Therefore, it appears that changes in the expression patterns of the developmental regulatory genes STM, WUS, and ARP have contributed to the unique determinate shoot development of Podotemaceae that allows them to grow in fast-flowing tropical rivers.
P0840 – ePoster
Molecular phylogeny and ecological evolution on
Daphne
(Thymelaeaceae)
Kawahara, T 1
Ohi-Toma, T 4
, Yamashita, N
, Murata, J 4
2 , Lei, T 3 , Miyaura, T 3 ,
1 Hokkaido Research Center, Forestry and Forest
Products Research Institute, Japan; 2 Kansai Research
Center, Forestry and Forest Products Research Institute,
Japan; 3 Faculty of Science and Technology, Ryukoku
University, Japan;
Tokyo, Japan
4 Botanical Gardens, University of
The genus Daphne (Thymelaeaceae) is a small shrub which occurs at understory or open meadow. In Japan, three different types of leaf phenology are recognized in the genus. The first type is evergreen ( D. kiusiana and D. miyabeana ); the second summer-green deciduous ( D. pseudo-mezereum var. koreana ); and the third wintergreen deciduous ( D. pseudo-mezereum subsp. pseudomezereum and subsp.
jezoensis ). Leaf phenology is closely related to ecological strategy in photosynthesis.
We reconstructed phylogeny based on DNA sequences of
ITS, rbc L, mat K, trn Ltrn F, psa B-rpl4 and trn Hpsb A spacers in order to examine directions in ecological evolution for Daphne . Molecular phylogeny revealed the evergreen species and the deciduous species are monophyletic respectively with high bootstrap probabilities. Winter-green deciduous species were derived from summer-green deciduous species. Daphne pseudo-mezereum complex composed of summer-green and winter-green deciduous taxa is monophyletic, and D. mezereum , summer-green species, is the sister.
Considering out-group character states, the phenology evolved from evergreen to winter-green deciduous by way of summer-green deciduous. Evergreen species have shade-tolerance and most of them occur in warmtemperate evergreen forest. Such ecological character of evergreen and physical one of effective use of week light through a year is adopted for the environment of understory in evergreen forest. Relatively high selfing rate of D. kiusiana which has gynodioecious flowers is

also adopted for less frequency of pollination. Summergreen species such as D. mezereum and D. pseudomezereum var. koreana often grow on calcareous lands.
The habitat is open and less competitive with other plants. In the habitat, D. pseudo-mezereum var. koreana has lost the character of shade-tolerance. Winter-green deciduous species, D. pseudo-mezereum subspp. pseudomezereum and jezoensis occur in deciduous forest. The habitat is on common forest soil and more competitive.
These species are also shade-intolerant and less adapted for summer environment of deciduous forest. Therefore it is supposed that change from summer-green to wintergreen deciduousness would be essential to make use of strong light effectively in deciduous forest with the physiological characters of shade-intolerance. Daphne pseudo-mezereum subsp.
jezoensis is gynodioecious, but has low self-compatibility. Since the pollinator fauna is rather rich in spring of deciduous forest, high pollination success by outcrossing there is more expected in evergreen forest. It means sexual differentiation has evolved in the genera. Despite D. miyabeana being one of evergreen members, it occurs in deciduous forest.
Since the habitat is located in heavy snowfall area in
Japan, the species is conserved from coldness and dry by staying in snowfall.
P0841 – ePoster
Dioecy vs. species separation: who wins? The genus
Struthanthus
as an example
Kazandjian, A 1
1 Universidad Simon Bolivar, Venezuela
Struthanthus is a large and diverse genus in the
Loranthaceae and is represented in the New World tropics by around 70 species. The genus is dioecious and is characterized by having opposite leaves, flowers about
2 to 8 mm long arranged in triads of three flowers set on a small cap formed by three basally-connate bracteoles.
Female and male plants can be distinguished by floral buds as male buds are oblanceolate and female buds are cylindrical. Within the genus, S. dichotrianthus Eichler
(1868), S. syringuifolius (Mart.) Eichler (1868) and S. phillyraeoides (Kunth) Blume (1830), are the most abundant species in Northern South America as is shown by the relatively large number of collections in local and foreign herbaria. Struthanthus dichotrianthu s has been collected in Brazil, Colombia, Guyana, Suriname and
Venezuela, S. syringuifolius distributed in the same areas as S. dichotrianthus but it has not been collected in
Colombia and Trinidad and S. phillyraeoides is considered endemic from Venezuela. A detailed morphological examination of specimens revealed considerable inter- and intra-population variation in the characters used to identify S. syringifolius and S. dichotrianthus and to distinguish these last specimens from the ones identified as S. phillyraeoides . Most of the diagnostic characters used to circumscribe the species were not useful for identification. To objectively evaluate the identity of the three species, multivariate analyses were performed on a morphological data set of 28 characters based on 184 specimens, collected over a wide geographical range. These analyses did not separate the species but separated the specimens into two groups based on sex alone. When male and female subsets and vegetative characters were analyzed separately, the analyses were unable to separate S. dichotrianthus from
S. phillyraeoides . Struthanthus syringifolius were separated in two groups when analyzing female specimens, one grouped specimens with short bracteoles
(less than 2 mm long) and a second one with bracteoles up t o 5 mm . These results indicate that S. dichotriantus and S. phillyreoides cannot be distinguished morphologically based on the characters used and therefore these two species should be considered synonymous. Struthanthus syringifolius is proposed to be separated in two subspecies as the short bracteole specimens are located in Venezuela and northern Brazil and the long bracteole ones in southern Brazil.
P0842 – ePoster
Phylogeny of the tribe Cynoglosseae (Boraginaceae s. str.) as inferred from nrDNA ITS sequences in Iran
Esmailbegi, S 1
1
M 2
, Kazempour Osaloo, S 1 , Mirtadzadini,
Dept of Plant Biology, Faculty of Biological Sciences,
Tarbiat Modares Universit, Iran; 2 Dept of Biology,
Faculty of science, Shahid Bahonar University, Iran
Boraginaceae s. str. (=subfamily Boraginoideae) comprises some 1600 species in approximately 100 genera and have their center of diversity in Eurasia. In
Flora of Iran, the family comprises 36 genera and over
180 species. The tribe Cynoglosseae includes 11 genera and over 50 species in Iran, being one of six traditionally recognized tribes. Paracaryum is the largest genus of the tribe in our area. A total of 63 accessions including 39 species of Cynoglosseae (including 12 species) along with 20 species of tribe Eritrichieae and two species of
Echiochilon (Echiochileae) plus Heliotropium bacciferum and Tournefortia rubicunda as outgroups was included in the phylogenetic analyses using nrDNA ITS sequence data. This region was amplified using appropriate primers by PCR and then PCR products were sequenced by the BigDye terminator cycle sequencing kit in an ABI Prism 3730xl DNA Analyzer. The sequences were aligned using Muscle program. The phylogenetic reconstruction was conducted using maximum parsimony, maximum likelihood and Bayesian methods as implemented in PAUP*/TNT, Tree-Puzzle/GARLI and Mrbayes softwares, respectively. The present data revealed that Cynoglosseae at the current status is not monophyletic, its taxa were intermixed with members of
Eritrichieae. Trichodesma with Caccinia and Heliocarya formed a sister group relationship. Omphalodes is allied with Myosotidium hortensi a of Eritrichieae. The remaining cynoglossoid taxa were gathered in a large polytomy. Within this polytomy, 12 recognized
Paracaryum species did not form a monophyletic group, some members of it formed several subclades and P. cynoglossoides was well allied with
Microparacaryum / Lepchiniella . Solenanthus was monophyletic and sister to Trachelanthus . Rindera with two species analyzed here was not appeared to be monophyletic. Cynoglossum with the inclusion of
Lindelofia longiflora was monophyletic.
605

P0843 – ePoster
A preliminary phylogenetic study of
Acantholimon
Boiss. (Plumbaginaceae) based on nrDNA ITS sequences in Iran
Moharrek, F 1 , Kazempour Osaloo, S 1 , Assadi, M 2
1 Dept of Plant Biology, Faculty of Biological Sciences,
Tarbiat Modares University, Tehran, Iran; 2 Dept of
Botany, Research Institue of Forests and Ranglands,
Tehran, Iran
Acantholimon is the largest genus of Plumbaginaceae. It comprises ca. 150 species mainly distributed in East
Mediterranean to Central Asia. About 80 species of the genus are distributed in Iran alone, of which 65 are endemic. In this study, phylogenetic relationships among
23 species comprising 17 accession representing 15
Acantholimon species, two Psylliostachys species, one
Armeria plus three Limonium species as outgroups were included in phylogenetic analyses using nrDNA ITS sequences. nrDNA ITS was amplified with appropriate primers by PCR and purified PCR products were sequenced using Big Dye terminator cycle sequencing ready reaction kit on an automated DNA sequencer.
Sequences were aligned with Muscle program and adjusted manually. Phylogenetic reconstruction was performed using maximum parsimony method with heuristic search and TBR branch swapping as implemented in PAUP* and Bayesian inference with
GTR+G evolutionary model by MrBayes. The analyses revealed that Acantholimon is well allied with
Psylliostachys and Armeria . The genus itself is monophyletic and comprising two subclades: One subclade is composed of A. aspadanum, A. gilliatii, A. flexosum A. cupreo-olivascens, A. bromifolium and A. bracteatum and the second one comprises A. festucaseun,
A. quniquelobum, A. erinaceum, A. cymosum, A. pterostegium, A. scirpinum, A. alavae, A. chlorostegium and A. kermanense . Sections Staticopsis, Glumaria ,
Acantholimon and Tragacanthina appeared to be nonmonophyletic, whereas the section Pterostegia with two species is monophyletic .
P0844 – ePoster
A phylogenetic study of the tribe Astereae
(Asteraceae) as inferred from nrDNA ITS sequences
In Iran
Jafari, Farzaneh 1
1 Dept of Plant Biology, Faculty of Biological Sciences,
Tarbiat Modares University, Tehran, Iran
A molecular phylogeny of the tribe Astereae
(Asteraceae) with an emphasis on the genus
Psychrogeton in Iran was studied. About 39 species in 15 genera of the tribe are distributed in Iran. A total of 108 accessions representing 38 genera of Astereae and five outgroups species from the relatives tribes (Anthemideae,
Gnaphalieae, Inuleae and Calenduleae) were included in phylogenetic analyses using nrDNA ITS sequences.
Maximum Parsimony method as implemented in PAUP*
4 and Bayesian inference using MrBayes were employed for phylogenetic analyses. The analysis of nrDNA ITS
606 showed that the tribe Astereae is monophyletic. Aster ,
Psychrogeton and Lachnophyllum appeared to be nonmonophyletic. Whereas Chamaegeron , Eurasian
Erigeron and Myriactis are monophyletic. Callistephus chinensis and Brachyactis roylei with the fertile achene of disc flower were nested among Psychrogeton species.
Psychrogeton nigromontanus was sister to a subclade including Ps. aucheri, Ps. aellenii, Ps. cabulicus, Ps. amorphoglossus, Ps. alexeenkoi and Ps. chionophilus .
Furthermore Ps. amorphoglossus, Ps. alexeenkoi and Ps. chionophilus were the closest relatives. The Galatella group is composed of Tripolium , Crinitaria and
Galatella . The current analysis like previous works demonstrated that Bellis was sister to the Galatella group. Conyzanthus squamatus is nested within
Symphiotrichum species and Dichrocephala integrifoli a is well allied with Grangea and Nidorella .
P0845 – ePoster
1
Evolutionary relationships in
Fritillaria
(Liliaceae)
Kelly, LJ
MF 1
1, 2
, Hill, L 4
, Berger, MR
, Leitch, IJ 1
Durham, UK; 4
3 , Day, PD
University of London, UK; 3
1 , Leitch, AR
Royal Botanic Gardens Kew, UK; 2
2 , Fay,
Queen Mary
Durham University,
Petersham Lodge, Richmond, UK
Fritillaria (Liliaceae) contains circa 140 species of bulbous perennials, which occur in temperate regions of the Northern Hemisphere. The genus includes taxa of horticultural importance, e.g. F. imperialis and F. persica , as well as those used in traditional medicine, e.g.
F. cirrhosa . Eight subgenera were recognised in the most recent classification of Fritillaria , with some of these further subdivided into sections and series. This classification was broadly supported by the results of a previous phylogenetic study based on DNA sequence data from two plastid regions and the nuclear ribosomal
ITS region, with species from three of the subgenera that were represented by multiple species forming monophyletic groups. Nevertheless, the largest subgenus,
Fritillaria , appeared polyphyletic as the result of a single species ( F. pallidiflora ) falling as sister to subgenera
Korolkowia, Petillium and Theresia in a clade separate to that of the other members of subgenus Fritillaria . Only
37 of the circa 140 species were available for inclusion in the previous work and, as well as contributing to current understanding of species evolution within the genus, the availability of a robust phylogenetic hypothesis for
Fritillaria is essential to provide an evolutionary framework for wider studies of the genus, such as research currently being conducted on genome size evolution. We present the latest results from an ongoing molecular phylogenetic study aimed at elucidating relationships between all species of Fritillaria . To date, sequence data have been obtained from >120 individuals, representing >85 species. Our results confirm the polyphyletic nature of subgenus Fritillaria , with species from this group falling within two distinct clades; a main subgenus Fritillaria clade is recovered (comprising around 45 of the species sampled so far), whilst samples from Central Asia and China (eight of the species sampled so far) are placed within a distinct part of the tree. Moreover, all members of the endemic Japanese subgenus Japonica , represented in previous phylogenetic

work by a single individual, have now been sequenced, allowing insights into the evolution of these species.
P0848 – Poster
Botanical illustration in Vienna once and now – exemplified on plants from the Pannonian region
Kiehn, M 1 , Hudler, P 2 , Pertl, M 3
1 Botanical Garden, University of Vienna, Vienna,
Austria;
Austria;
Ireland
2 Zoologisch–Botanische Gesellschaft, Vienna,
3 6 St David's Terrace Glasnevin, Dublin,
Botanical illustrations are a very effective way of passing on information about plants. In Vienna, skillfully laid-out plant portraits play an important role in scientific publications and florilegia since the 18th century. This can, e.g., be prominently seen in the work of Nikolaus
Joseph von Jacquin, the famous botanist and director of the Botanical Garden of the University of Vienna.
Jacquin specifically trained and selected plant illustrators to work for him. Thus, one of his masterpieces, the
Florae Austriacae, sive plantarum selectarum in Austriae archiductu: sponte crescentium icones, ad vivum coloratae, et descriptionibus, ac synonymis illustratae , contains 500 plates with coloured copper engravings of plant portraits. They are mostly done by the artist Franz
Scheidl (Lack 2001: Ein Garten Eden ) and illustrate
Jacquins’s Latin descriptions of species native to the surroundings of Vienna. Numerous plants shown and described in this work are typical elements of the
‘steppe-vegetation’ of the xerothermic regions southeast of Vienna, the so called ‘Pannonian’ flora. Preserving rare species of this floristic region is a central aspect of the Botanical Garden of the University of Vienna today.
While the botanical illustration remained an important tool in descriptive botany, the tradition of training artists in the special skills needed for high quality plant portraits faded away in Vienna. In recent years however, plant illustration activities are booming again worldwide.
Consequently, the Austrian Zoological-Botanical Society
(ZOOBOT) started to organize training courses in cooperation with the Botanical Garden of the University of Vienna (HBV). The courses are guided by Margareta
Pertl, who was born and trained as an artist in Vienna, and now lives and teaches in Dublin. She has been working with the HBV for many years, doing plant portraits – mostly of orchids. Her plant illustrations have been awarded medals by the Royal Horticultural Society and one of her orchid portraits was exhibited at the 13th
International Exhibition of Botanical Art & Illustration in the Hunt Institute for Botanical Documentation,
Pittsburgh, PA – an institution dedicated to collecting high quality contemporary plant portraits. To link to the
Viennese tradition of botanical illustration as art form connected to science, only plants from the HBV are used as motifs for botanical illustrations. This way, a 'new florilegium' of the HBV-collections is going to be developed over the years, with a main focus on plants from the ‘Pannonian region’. In addition to the artistic aspect, these illustrations are contributing to the scientific and educational work done at the HBV: in autumn and winter, copies of the illustrations are exhibited in the garden to create curiosity and to offer an opportunity to perceive plants from a different point-of-view. The plant portraits will also be used to supplement texts for a planned guidebook related to the conservation efforts at the HBV. Historical and contemporary artwork can be conpared based on plant portraits of the same species from the 18th century and from the recent florilegium done by students in the HBV .
P0849 – ePoster
Geographic distribution and natural habitats of diploid and triploid
Lilium lancifolium in east Asia
1
Kim, J-H 1 , Song, Y-S 1
Hiramatsu, M 3
, Kim, N-S 1 , Xuan, YH 2 ,
Kangwon National University, Korea;
University, China; 3 Kyushu University, Japan
2 Yangzhou
Diploid and triploid forms of tiger lily ( Lilium lancifolium , synonym L. tigrinum ) are known. Because the exact geographic distribution of each ploidy has not been attempted, we explored the geographic distributions of the tiger lilies extensively in Korea, China, and Japan.
In Korea, the triploids were found both inland and coastal or islands in Yellow Sea, but the diploids were not present inland and found exclusively in the islands and coastal region of the Yellow Sea. We also surveyed their distributions in coastal regions of north eastern China and coastal regions in Japan that face toward Korea. In the Chinese coastal region, triploids were only found, but not diploids. In Japan, there were no diploids in the western coast region facing Korea, but they were present in the islands of Stushima, Hirada, and Goto of Kyushu province. From the facts that wide distribution and very high genetic diversity of the diploids in the Korean islands in Yellow Sea, we posit that centre of the origin of the diploid tiger lily may be somewhere on these islands.
P0850 – ePoster
Synoptical molecular phylogeny of Liliales insight from the combined data of three plastid loci,
mat
K, rbc
L and atpF-H
Kim, JS 1 , Hong, J-K 1 , Chase, MW 2 , Kim, J-H 1
1 Kyungwon University, Korea; 2 Royal Botanic Gardens,
Kew, UK
Many molecular studies showed that Liliales is a wellsupported monophyletic group defined by synapomorphy of the tepal nectaries and extrorse anthers, in contrast to the septal nectaries and introrse anthers commonly found in other monocots. It is composed of up to 1,500 species, over 67 genera and 9–11 families. Their distributions are generally known to two patterns as widely in northern hemisphere or narrowly in southern hemisphere. In
Korea, about 50 species, 18 genera of 4 families,
Colchicaceae, Liliaceae, Melanthiaceae and Smilacaceae, are occurred. In this study, we try to define the inter- and infra-familial relationship among Liliales by the molecular phylogenetic analyses based on three plastid loci, mat K, rbc L and atp F-H. Intergenic spacer of atp F-H is newly applied for Liliales. The result of phylogenetic analyses of 88 species, 47 genera, of 9 families including all of Korean Liliales strongly supported the monophyly
607

of Liliales and all of 9 families with 97 BP and 100 BP, respectively. From the strict consensus tree based on the combined data, it was clear the familial relationships in
Liliales. It is distinctly divided to two major clades. The one was well supported by 100 BP and composed of large Liliaceae and a clade of three small families,
(Rhipogonace+Philesiaceae)+Smilacaceae, and the other, although which is weakly supported by 63 BP, included a large family Colchicaceae and a clade supported by 87
BP of
(Melanthiaceae+(Alstroemeriaceae+Luzuriagaceae))+Ca mpynemataceae. Among the families, it was clearly confirmed the close relationship supported by 100 BP between Rhipogonaceae and Philesiaceae as well as between Alstroemeriaceae and Luzuriagaceae. Besides, it was well supported and explained the infra-familial relationship within three larger families of Liliaceae,
Melanthiaceae and Colchicaceae. However, the genus
Aletris which has been placed in family Melanthiaceae by APG II (2003) needed to be recircumscribed into family Nartheciaceae. We are planning to analyze more plant materials including family Petermaniaceae and
Corsiaceae not investigated in this study and more molecular markers for constructing a new circumscription about Liliales reflecting the phylogenetic relationship in the future.
P0851 – ePoster
DNA analysis identifies roots infiltrating the lava
1
3 tubes in Jeju World Natural Heritage of Korea
Kim, S 1 , Paik, W 2 , Heo, K 3 , Jeon, Y 4
Sungshin University, Korea;
, Suh, Y 5
2 Daejin University, Korea;
Kangwon National University, Korea;
Self-Governing Province Office;
4 Jeju Special
5 Seoul National
University, Korea
Geomunoreum Lava Tube System of Jeju Volcanic
Island in Korea has been inscribed in the list of
UNESCO World Natural Heritage due to its unrivaled spectacular beauty . After the caves had been formed by lava flow, they have undergone the calcite formation such as soda straws, stalactites and columns . The addition of characteristics of limestone caves to the lava caves has created unprecedented rarity as well as dramatic landscape. The shell sand (calcite dune deposits) on the ground above the caves, blown from the coast, is dissolved into calcium carbonate, which infiltrates the caves and forms the fascinating limestone cave features in addition to the landscape of lava tube . In the formation of limestone cave features, such as the straight-formed stalactites, plant roots play a crucial role because the rainwater sips through along the roots.
Stalactites grow along the roots, which will become petrified eventually . To understand the contribution of plant roots in forming cave landscape, we identified roots infiltrating the cave by comparative analysis of trn Hpsb A intergenic spacer located in chloroplast genome .
We examined the flora in the area above caves and set up a local DNA barcode system of trn Hpsb A . Sequences obtained from roots in the caves were compared and identified for species. Confirmed are Equisetum ramosissimum Desf., Pueraria lobata (Willd.) Ohwi,
Rosa maximowicziana Regel, Koelreuteria paniculata
Laxm., Elaeagnus umbellata Thunb., Ficus erecta
608
Thunb. var. sieboldii (Miq.) King, Setaria sp. and
Washingtonia filifere (L.Linden) H.Wen
. Washingtonia filifere has been recently introduced to Jeju Island, and often transplanted for the purpose of landscaping . The penetration of W. filifere roots through cracks could cause a serious problem because its root found in the ceiling of a cave was considerably thick . The outgrowth of its thick roots will certainly accelerate the collapse of cave ceiling . DNA data strongly suggest that W. filifere should not be planted to conserve caves in the area . In addition, we have found the massive penetration of E. ramosissimum roots in a couple of caves . It is surprising that roots of Equisetum , an herbaceous plant, can develop such a massive formation to reach deep into the cave ceiling, which is at least 4–5 m deep . The cultivation in the field above the caves has been prohibited at least one year after the Lava Tubes were inscribed in World
Natural Heritage sites in the summer of 2007 . In the past, periodic plow to cultivate crops above ground must have prevented plants from developing roots to penetrate into the caves. In results, DNA analysis has provided useful information not only to understand the roles of plants in the process of geological formation, but also to conserve the environment of the Lava Tubes in Jeju World Natural
Heritage in Korea.
P0852 – ePoster
Floral morphology and anatomy of
Rauvolfia
L.
(Apocynaceae, Rauvolfioideae), with emphasis on
Kinoshita, LS 1
1 secretory structures
, Simões, AO 2 , Koch, I 3 , Castro, MD 1
Universidade Estadual de Campinas, Brazil; 2
Universidade de São Paulo/Campus Leste, Brazil; 3
Universidade Federal de São Carlos/Campus Sorocaba,
Brazil
Rauvolfia L. is the largest genus of Vinceae
(Rauvolfioideae), and comprises about 60 species distributed throughout the tropics. Despite morphological variation in reproductive traits within the genus, there are a number of diagnostic features, such as whorled branches and leaves, drupaceous fruits and pollen grains with a discernible thickening on both sides of each colpus. Its monophyly has been strongly supported in several phylogenetic studies, even though most of the traditionally recognized infrageneric ranks and polymorphic species with uncertain delimitation (such as
R. tetraphylla ) were found to be paraphyletic. In the present study, the floral morphology and anatomy of
Rauvolfia species ( R. balansae, R. ligustrina, R. sandwicensis, R. sellowii, R. sprucei, R. sumatrana, R. tetraphylla, R. vomitoria, R. weddelliana ) and two related taxa from Vinceae ( Ochrosia compta, Vinca minor ), were analyzed in order to identify characters with potential taxonomic and evolutionary value. The selected species of Rauvolfia encompass the morphological, geographical and taxonomical diversity of the genus. A nectary ring surrounding the ovary and the style insertion into a depression at the ovary apex were observed in the nine species of Rauvolfia and thus represent putative sinapomorphies of the genus. The style-head is an organ of the gynoecium formed by two apical appendages, a main body covered by secretory cells, and a basal ring consisting of epidermal cells oriented downwards. It

provides a substantial number of underexplored characters with taxonomic value. The shape of the stylehead main body and its pattern of coverage by secretory cells, apical appendage morphology and the size of the basal ring are highly variable in the species analyzed here. Nevertheless these features are more constant among closely related species of Rauvolfia , that they are taxonomically relevant. A group of strongly stained cells on the calycine lobe margins was noted in six species ( O. compta, R, ligustrina, R. sprucei, R. sumatrana, R. tetraphylla, V. minor ). This group of cells resembles the calycine colleters that have already been reported in other species of Apocynaceae although they lack a discernible stalk and a secretory head. Further studies in other species of Rauvolfia and related genera of Vinceae may shed a light on the evolution of calycine secretory structures within Apocynaceae.
P0853 – ePoster
1
Getting the most out of pathway keys in a web environment
Klazenga, N 1 , Thiele, K 2
Royal Botanic Gardens Melbourne, Australia; 2 Western
Australian Herbarium, Australia
In some ways pathway identification keys, such as the traditional dichotomous keys, are superior to matrix keys.
It is possible to put a lot more expert taxonomic knowledge into pathway keys – as the author of a key, rather than an algorithm, can tell users what characters to look at and into what order – and people tend to learn a lot more about a group of organisms by repeatedly using a pathway key than by using a matrix key. However, pathway keys do not work nearly as well as matrix keys in electronic identification systems. Creating a key to a subset of taxa in a matrix key is a rather trivial exercise, while doing the same thing for a pathway key is about as challenging as making a matrix key work on paper. We endeavour to develop a procedure that will enable people to create on-the-fly keys to any subset of taxa from a pathway key that is stored as a database of key nodes
(couplets) and key leads (alternatives). The subsets of taxa may be based on any criterion, such as occurrence in a geographical area, a shared morphological character or combination of characters, higher taxon affiliation, etc.
Such a procedure would not only enable us to, for instance, easily create keys for local floras from keys of regional floras, but will also enable toggling between matrix and pathway keys and will go a long way to enable us to adapt staged keys – keys divided into keys to families, genera and species – to a new classification. We discuss both the benefits and limitations of such an approach to making pathway keys more flexible and powerful.
P0854 – ePoster
A combined phylogenetic reconstruction of the
Podocarpaceae based on
rbc
L, nrITS1 and NEEDLY intron2 sequence as well as anatomical, morphological and biogeographical data
Knopf, P 1 , Schulz, C 1 , Stuetzel, T 1
1 Institute for Evolution and Biodiversity of Plants, Ruhr
University Bochum, Germany
The Podocarpaceae family consists of 19 genera, 194 species and 9 varieties and is the second largest gymnosperm family. The species are predominantly distributed in tropical and subtropical montane habitats from Central America, South America, Africa (including
Madagascar) and Indochina through Malesia to Australia and Polynesia. Thus, the family shows a strong
Gondwanan relationship like the Cunoniaceae and
Nothofagaceae. Nearly all Podocarps are characterized by an epimatium covering the seed and a swollen, fleshy receptacle below the seed which is eponymous for this group. Studies of the last century described numerous new species or segregated taxa to new genera or families using anatomical and morphological characters.
However, the Podocarpaceae remain poorly investigated phylogenetically. Weakly supported groups or taxa which had uncertain positions were shown in previous molecular phylogenetic analyses of Podocarpaceae. A comprehensive analysis of the Podocarpaceae on the species level using molecular data, biogeography, anatomy and morphology was still lacking. To address this need, the present study used sequence analyses of rbc L, nrITS1 and NEEDLY intron2 of over two thirds
(74.7%) of all Podocarpaceae species (183 accessions of
145 taxa) representing all genera except Parasitaxus De
Laub. Several disputed taxa ( Dacrydium pierrei Hickel,
Phyllocladus glaucus hort. ex Carrière, Podocarpus decipiens N.E.Gray, Podocarpus forrestii Craib and
W.W.Sm., Podocarpus hallii Kirk, P. oleifolius D.Don var. costaricensis J.Buchholz & N. E.Gray., and
Podocarpus philippinensis Foxw.) and the genus
Phyllocladus Rich. ex Mirb. were also included.
Furthermore, a comprehensive anatomical and morphological study ensured the accurate identification of the investigated taxa and provided features that support clades within the molecular analyses. Two reconstruction methods (Bayesian inference and parsimony (TNT 1.1)) were used to detect methodindependent monophyletic groups. Our results show three major clades within the Podocarpaceae: the
Prumnopytidioid, Dacrydioid, and Podocarpoid clade.
The Podocarpoid clade consists of the Polypodiopsis clade (genera A frocarpus, Nageia and Retrophyllum ) and
Eupodocarpus (= Podocarpus L’Her. ex Pers.) clade. In total, 20 new prominent groups, including 10 new monophyletic groups within the formerly poorly resolved genus Podocarpus L’Her. ex Pers., were discovered. The subgenus Foliolatus De Laub. consists of the Fijian-,
Indochinese-, Neriifolius-, New Caledonian-, and the
Southern Malesian subclade. The African-, Australis I-,
Australis II-, subtropical American and the tropical
American subclade were detected within subgenus
Podocarpus De Laub. Anatomical and morphological features support the resolved topology as well as the geographical distribution does . The monotypic
Sundacarpus C. N. Page is placed back within
Prumnopitys Phil. to avoid a paraphyletic genus
Prumnopytis. The Phyllocladaceae family is subsumed into the Podocarpaceae again.
P0855 – ePoster
Late Middle Miocene vegetation based on plant macrofossils from the Kanto Plain, central Japan
609

1
Kobayashi, M 1 , Okitsu, S 1
Graduate School of Horticulture, Chiba University,
Japan
Late Middle Miocene Yagii Formation (about 12–9Ma), the Kanto Plain, central Japan contains many plant macrofossils. Although the fossil leaf assemblages were described in detail by Ozaki (1991), carpological assemblages have not been studied. We studied fruits and seeds fossils from the formation to reconstruct spatial distribution of plant habitat and vegetation. The mudstone, lignite and sandstone of the Yagii formation contains many plant macrofossils. The mudstone and lignite can be characterized as autochthonous. Their macrofossils indicated the vegetation in the lowland near the fossil bearing site. The sandstone can be characterized as allochthonous. Its macrofossils indicated the vegetation of the mountain slope or valley. The fruits and seeds assemblage from the mudstone and lignite contained many fossil leaves, seeds and cones of
Metasequoia , leaves of Glyptostrobus , fruits of
Potamogeton , and seeds of Proserpinaca. Metasequoia,
Glyptostrobu s and Proserpinaca are now extinct from
Japan. Metasequoia and Glyptostrobus are now distributed only in Asia but they were common in the
Northern Hemisphere in Neogene. Proserpinaca is now endemic to North America but its fossils have been found from European Neogene. The fossil record from the
Yagii formation is the first record from Asia. The fruits and seeds assemblage from the sandstone contained various tree taxa, cf. Fagus crenata, Eurya, Stewartia,
Styrax, Meriosma, Euptelea, Carpinus, Paulownia,
Callicarpa, Viburnum and Liquidambar and also herb taxa, Sambucus chinensis, Patrinia, Viola , and Carex .
Fagus crenata is one of the dominant species in Japanese slope forest. Carpinus, Stewartia, Styrax, Meliosma and
Callicarpa are now common deciduous broad-leaved trees and Eurya is common evergreen broad-leaved tree in Japanese slope forest. Euptelea is distributed only in
Japan and China. Its habitat is characterized as mountain valley or the place where landslides occasionally take place. Liquidamber is now extinct from Japan but common in Japanese Neogene flora. Based on those plant macrofossils, around the Kanto Plain including mountain slopes, we reconstructed the palaeovegetation.
Coniferous forest dominated by Metasequoia was distributed in lowlands like fluvial back marsh because most of the samples from the mudstone and lignite contained a lot of Metasequoia fossils. Leaf assemblages also suggested Metasequoia was dominated in wetland and riparian vegetation (Ozaki, 1991). Glyptstrobus and
Proserpinaca were distributed in more wet area than
Metasequoia in this forest. Deciduous forest dominated by Fagus were distributed in the slope with higher elevation. Euptelea were distributed in mountain valley or landslide area. In the late Middle Miocene lowlands were important habitat of extinct species from modern flora of Japan such as Metasequoia , Glyptostrobus and
Proserpinaca while slope area were important habitat of ancestor of the deciduous broad-leaved trees such as
Fagus, Euptelea and Carpinus which are now common deciduous tree in Japan. Palaeovegetation reconstructed by our study suggests that plants in lowlands were affected by sea-level fluctuation and sedimentation, causing the extinction and plants survived in slope or mountain area existing in relatively steady environment where landslides occasionally took place.
P0856 – ePoster
Chloroplast genome sequencing of the moss,
Andreaea nivalis
Kosugi, I 1 , Shimamura, M 1 , Yamaguchi, T 1 , Deguchi, H 1
1 Dept. of Biol. Sci., Graduate School. of Sci., Hiroshima
University, Japan
One of the basal groups of mosses, the class
Andreaeopsida is clearly distinguished from other groups in having characteristic sporophyte morphology (e.g. capsule splitting longitudinally in four slits, no stalk, and no peristomes). Here we report a chloroplast genome sequence of Andreaea nivalis based on pyrosequencing.
This information enabled us to compare the genome structure of Andreaeopsida with that of the other three major taxonomic groups of mosses, Takakiopsida,
Sphagnopsida and Bryopsida. The cpDNA fragments were amplified by use of long and acculate polymerase chain reaction (LA-PCR) and entire cpDNA were covered with 15 partially overlapped PCR fragments, which are approximately 6–12 kb in length. These PCR fragments were pyrosequenced using the GS FLX system. The obtained reads were assembled and remaining gaps were closed by individual PCR reaction and nucleotide sequencing. Eventually sequence data were assembled as a circular double-stranded DNA molecule. Chloroplast genome of A. nivalis had a common quadripartite structure in land plants: a pair of
IRs separated by the LSC and SSC regions. The genome contains at least 77 proteins, 32 tRNA, 4 rRNA, 7 open reading frames (ORFs) and some pseudogenes. The ribosomal protein small subunit 16 (rps16) which is unknown from Bryopsid mosses, is found from the chloroplast genomes of A. nivalis and another primitive moss Takakia lepidozioides . On the other hand, some functional genes (e.g. cysA and cysT) known from
Takakia seem to be pseudogenized in Andreaea due to insertion of stop codon and frame shift. We also found a pseudogene of tufA in A. nivalis that shares 70% nucleotide similarity with the chloroplast encoded tufA gene of Chara vulgaris . Because tufA is located in nuclear genome in land plants, but in chloroplast genome in green algae, it has been completely transferred from the chloroplast genome to the nuclear genome through the evolution of land plants. The pseudogene in chloroplast genome of A. nivalis seem to be a relic of functional gene in chloroplast genome. The tufA like sequence is also found from other primitive mosses, T. lepidozioides and Sphagnum palustre but absent from bryopsid mosses. The tufA like sequence of Takakia and
Sphagnum share some common indel patterns (deletions and insertions) that do not exist in Andreaea . These results might reflect the intermediate phylogenetic position of Andreaea between pivotal groups ( Takakia and Sphagnum ) and more advanced groups (bryopsid mosses).
610

P0859 – ePoster
The phylogeny, biogeography and morphological evolution of
Gaultheria
(Ericaceae) from Australia and New Zealand
Kron, K 1 , Bush, C 1 , Wagstaff, S 2 , Fritsch, P 3
1 Wake Forest University, USA; 2 Allen Herbarium,
Landcare Research, Lincoln, New Zealand; 3 California
Academy of Sciences, San Francisco, California, USA
Phylogenetic relationships within Gaultheria L. from
Australia and New Zealand were examined by using
DNA sequence data from matK, ndhF, nrITS, waxy and lfy. In the combined parsimony and maximum likelihood analyses, all Australia/New Zealand species form a clade that is sister to a clade of temperate South American species. However, Australian and New Zealand species did not form clades exclusively corresponding to geographic regions. Optimisation of morphological characters that have been emphasised in classifications of
Gaultheria onto the molecular phylogeny revealed that, within the Australia/New Zealand clade, non-fleshy fruiting calyces, berries and solitary-flowered inflorescences each evolved twice, from fleshy fruiting calyces, capsules and multiple-flowered inflorescences, respectively. Geographic regions mapped onto the phylogeny suggests a South American origin of the
Australia/New Zealand clade, followed by three dispersal events from New Zealand to Australia. Whether the origin is from temperate or tropical South America is ambiguous in our analysis.
P0861 – ePoster
Bryophytes captured from the atmosphere
Kubo, H 1 , Handa, S
Tsubota, H 4
2 , Nakahara-Tsubota, M 3 , Mukai, S 4 ,
1 Dept of Biological Science, Graduate School of Science,
Hiroshima University, Japan; 2 and Health Association, Japan;
Hiroshima Environment
3 Cooperative Research
Fellow of Natural History Museum and Institute, Chiba,
Japan; 4 Miyajima Natural Botanical Garden, Graduate
School of Science, Hiroshima University, Japan
Bryophytes are the second largest group of land plants, and they occur in almost all terrestrial habitats, with the exception of saline waters, on every continent.
Bryophytes are propagated naturally by sexual and/or asexual reproduction. Bryophyte plants, their sexual spores and asexual diaspores, such as gemmae or other specialized propagules, are generally small. Spores are thought to be important for long-distance dispersal to colonize newly available habitats, while gemmae and other propagules are thought to generally serve as shortdistance dispersal agents and serve mainly to maintain local populations. Since the late 1800s, dispersal ability or dispersal potential of bryophytes (i.e., number, size and transportability of diaspores, their physiological susceptibility – such as to UV radiation) – have been studied by many authors. The mechanisms and patterns of dispersal are reflected in the genetic composition and structure of populations, and in their speciation. The dynamics of dispersal of bryophytes in aerobiology, however, has been little studied, especially dispersal distance and frequency of capture of microscopic diaspores. In the present study, we captured, cultivated and then sequenced DNA from aerial materials, such as rain drops and snows. DNA barcoding of the cultured materials indicated both long-distance and short-distance dispersal originating from sources outside Japan. The results support the concept of long-distance dispersibility in bryophytes and the use of genetic markers in identification of the species and putative origin of airborne propagules.
P0863 – ePoster
Phylogeography of genus
Deparia
(Woodsiaceae)
1
Kuo, L-Y 1
Wood, K 4
, Ebihara, A
, Ranker, T 5
2 , Shinohara, W 3 , Kato, M
, Wang, C-N 1 , Chiou, W-L 6
2
2 ,
Institute of Ecology and Evolutionary Biology, National
Taiwan University, Taiwan; Dept of Botany, National
Museum of Nature and Science, Japan; 3 Dept of Botany,
4
Graduate School of Science, Kyoto University, Japan;
National Tropical Botanical Garden, USA;
Botany, University of Hawai`i at Manoa, USA; 6
5 Dept of
Division of Botanical Garden, Taiwan Forestry Research
Institute, Taiwan
Deparia is a fern genus in Woodsiaceae (sensu Smith et al., 2006, 2008), comprising around 70 species and mainly distributed in Asia, while several species are distributed and endemic in the Hawaiian Islands, North
America, and Africa. We conducted a phylogenetic analysis of DNA sequence data and found support for seven major clades within Deparia . In each clade, the
East Asian + Southeast Asian region had greater diversity and/or species richness than the other regions.
According to their distribution, these clades could be further identified as either oceanic or continental. Basal clades were composed of continental taxa while derived clades were oceanic taxa. Molecular dating analyses were undertaken to estimate the divergence times of all clades.
The results suggested that Deparia taxa dispersed from
Asia, and then colonized in Pacific islands, North
America, and Africa during the Pliocene. In addition to these ancient dispersal events, there were some recent expansions in oceanic groups, and the cytogeographical pattern of these oceanic groups implied that polyploidization may play an important role in their expansion.
P0864 – ePoster
Reproductive structures
Zlatkocarpus
,
Myricanthium and their associated foliage from the Cretaceous of
1 central Europe
Kvacek, J 1
National Museum, Prague, Czech Republic
Cretaceous angiosperms from the Peruc–Korycany
Formation (Cenomanian of the Czech Republic, Central
Europe) show high diversification. More than 100 species of angiosperms are described from there.
Zlatkocarpus pragensis, Z. brnikianus and Myricanthium amentaceum are introduced here as compound
611

inflorescences/infructescences consisting of primary axes bearing semi-decussately arranged spikes. Each spike has helically arranged unicarpellate and unilocular fruits. consisting of unilocular fruits with sessile stigma.
Zlatocarpus as better preserved shows Each fruit apparently containing a single, orthotropous seed. The stigma is indistinct and sessile at the apex. The fruit wall has distinct globular protrusions interpreted as resin bodies. The fruits are supported at the base by a small floral cup and a bract. Sessile stigma bears pollen similar to dispersed pollen broadly referred to the extinct pollen genus Retimonocolpites. inflorescences/infructescences of
The
Zlatkocarpus –
Myricanthium type are interpreted as members of broader chloranthoid clade. The intepretation is based on their semidecussate arrangement of secondary axes, unilocular fruits, sessile stigma and presence of Retimonocolpites type of pollen. Foliage of Debeya -type ( Debeya coriacea – Myricanthium amentaceum, Debeya brnikensis – Zlatkocarpus brnikensis, Debeya aralioides –
Zlatkocarpus pragensis ) occur repeatedly in several localities in association inflorescences/infructescences of with the
Zlatkocarpus –
Myricanthium . Both foliage and inflorescences/infructescences contain the same kind of resin bodies. The leaves show venation, chloranthoid teeths and stomatal aparati remarkably similar to the living representatives of the family Chloranthaceae .
P0868 – ePoster
Acalypha herzogiana
(Euphorbiaceae), the correct name for an intriguing and commonly cultivated species
Levin, GA 1 , Steinmann, VW 2
1 University of Illinois, Champaign, USA; 2 Instituto de
Ecología, Centro Regional del Bajío, Pátzcuaro, Mexico
The genus Acalypha (Euphorbiaceae) contains three commonly cultivated ornamental species . Two of these,
Acalypha hispida (chenille plant) and A. wilkesiana
(copperleaf), have been propagated for centuries and are not known in the wild, although they are closely related to a group of southeast Asian and Pacific Island species .
A third species has only recently appeared in the horticultural trade, with the earliest evidence of its cultivation from the mid-1980s . This taxon is often grown in hanging baskets or as groundcover and owes its appeal to its showy terminal pistillate inflorescences, giving rise to its English vernacular names strawberry foxtail, red firetail, dwarf cattail, trailing red cattail, kittentail, strawberry firetail, dwarf chenille, and trailing chenille plant . It is frequently called A.
‘repens,’ which is not validly published. Other names used for this plant include A. hispaniolae and A. reptans (both synonyms of
A. chamaedrifolia ), as well as A. pendula , but it differs morphologically in significant ways from all of these .
Instead, it strongly resembles A. herzogiana , a native of
Argentina, Bolivia and Paraguay, with which it also shares identical ITS sequences . Careful examination of the cultivated plants reveals that they differ from A. herzogiana plants in the wild only by possessing dimorphic pistillate inflorescences, a feature otherwise unknown in Acalypha , and by lacking staminate inflorescences . One of these inflorescence types is
612 identical to the terminal pistillate inflorescences of wild
A. herzogiana , which are sessile, relatively inconspicuous, and present the typical structure of the majority of Acalypha in that the pistillate flowers are closely enveloped by accrescent foliaceous bracts and are not densely clustered. By contrast, the other inflorescence type is showy and highly anomalous . It consists of hundreds of densely clustered, ebracteate, abortive pistillate flowers . These unusual inflorescences resemble the staminate inflorescences of wild A. herzogiana in that they are terminal, pedunculate, and bear flowers in glomerules (as are staminate flowers in all Acalypha ), but the flowers have no ovaries, instead producing 5 to 8 styles that arise directly from the receptacle in the position normally occupied by stamens in staminate flowers . We conclude that these unusual flowers are homologous to the staminate flowers of the wild plants and are the result of a homeotic mutation .
Furthermore, we consider it unlikely that the homeotic mutation resulting in the development of the anomalous, showy pistillate inflorescences occurred in cultivation because without such inflorescences the plants would be non-descript and of little horticultural interest . The plant is a worthy subject for future research on homeotic mutations, floral development, and gene expression.
P0869 – ePoster
New records of mosses to Sichuan Province collected from Gongga Mountain
Li, ZH 1 , Cao, T 1 , Yu, J 1
1 Shanghai Normal University, Shanghai, China
Sichuan Province, in China’s western hinterland, is known as the ‘Land of Abundance, the heaven of animals and plants’. Mt Gongga is the highest peak in Sichuan. It is one of 25 key biodiversity regions in the world, studying of which means much to biodiversity conservation. But its richness of mosses has never been studied until now. Based on identification of more than
1000 specimens, 319 species belonging to 128 genera and 39 families of mosses are recognized. 22 species are new to Sichuan. According to the results of the analysis, the mosses there are very unique and diverse.
P0870 – ePoster
Studies on the role of LEAFY COTYLEDON1 gene in evolutionary innovation of seed program
Li, X 1 , Rao, G-Y 1 , Bai, S-N 1
1 Peking University, China
Bryophytes, pteridophytes, gymnosperms and angiosperms are recognized as four main groups of the land plants. In the Plant Tree of Life, pteridophytes are in a pivotally evolutionary position between bryophytes and seed plants because of their prominent alternation of the more or less balanced gametophyte and sporophyte generations of the life cycle. Their sexual reproduction produces single-cell spores but not yet seeds. Both gametophyte and sporophyte of fern can live independently. The gametophyte has no but sporpophyte has vascular system for water transport so that

gametophyte can not grow into a high and large standing plant. With such special evolutionary features and phylogenetic position among land plands, fern, therefore, provides excellent research system for plant evolutionary developmental study. The fundamental difference of seed plants and non-seeding land plants are the development of their embryos. In seed plants, the embryos endure a developmental stage when they become desiccated and dormant. In contrast, embryos in bryophytes and pteridophytes develop continuously to grow into a sporophyte. There is a phenotype among the leafy cotyledon 1(lec1) mutant of Arabidopsis thaliana , which appear to have continuously growing embryo. An important and challenging question thus arises: whether the formation of seeds, an innovative character of seed plants, is correlated to the function of LEC1-type gene.
This study is carried out to answer this question. We first conducted morphological observation on the life cycle and the developmental phases of the fern plant Adiantum capillus-veneris . Then we detected the function of
AcLEC1 gene using this fern system in aims of interpreting the evolutionary developmental mechanism of seed traits. By optimizing the culture system and based on the synchronized culture and the morphology observation of the developing phase of gametopyte and sporophyte, we make controlling of the life cycles of A. capillus-veneris feasible and effective. This set up the possibility of A. capillus-veneris to be a model system of developmental study of fern. Furthermore, our results that AcLEC1 gene do not express during the stages from antheridium and archegonium to the embryo develop but had similar functions to the LEC1 gene when transformed into the lec1 mutant provide experimental basis for us to document our working hypothesis that there could be three essential factors for seed development .
P0871 – ePoster
Argentinian Phanerogamic flora: Asclepiadoideae
(Apocynaceae)
Farinaccio, MA 1 , Liede-Schumann, S 2 , Rua, GH 3
1
2
Universidade Federal de Sergipe, Argentina;
University of Bayreuth, Germany; 3 Facultad de
Agronomía, Universidad de Buenos Aires, Argentina
The Apocynaceae s.l. family has approximately 5000 species and 395 genera classified in five subfamilies. The family now includes the previously recognized
Asclepiadaceae, a large group composed of the subfamilies Periplocoideae, Secamonoideae, and
Asclepiadoideae that is distinguished morphologically by important aspects of the construction of the pollination apparatus, which may be considered a further elaboration from the more basal subfamilies Rauvolfioideae and
Apocynoideae (the Apocynaceae s. str.). Distinctive features of Asclepiadaceae include pollen dispersed in pollinia or tetrads that are provided with a translator composed of style-head secretions that have become solidified. Robert Brown, in 1810, divided Apocynaceae s.l. into two families, Apocynaceae s. str. (diagnosed by pollen in monads) and Asclepiadaceae (diagnosed by pollen in tetrads or pollinia). His classification was widely followed until studies appraising the phylogeny and systematics of the Apocynaceae s.l. have shown the
Asclepiadaceae is monophyletic but nested within
Apocynaceae. The Asclepiadoideae comprises ca. 3000 species, around 40% of them in the New World. The
Neotropics is one of the centers of diversity for
Asclepiadoideae. Thus, in the New World, where biodiversity is considerable, this study is of special importance to contribute to the knowledge of the flora of
Argentinean Asclepiadoideae. The objectives are: to provide a monograph on the Asclepiadoideae
(Apocynaceae) for the ‘ Flora Argentina ’ Projec, information on the species’ morphological and biological particularities and their geographic distribution; to correct and to update nomenclatural information for the taxa in Asclepiadoideae. In Argentina, approximately 25 genera with ca. 180 species are found, 58 of which are endemics. The majority of the species are climbers; other forms include erect or straggling shrubs or subshrubs.
They inhabit subtropical forests, open habitats and semiarid regions. They occur from 300 to 1500 m above sea level. The result will serve as a basis for various projects on the flora and the conservation of the areas of occurrence of these species as well as on the human adaptation and the conservation of biosphere resources.
CONICET, IAPT.
P0872 – ePoster
The genus
Croton
L. (Euphorbiaceae) of southeast
1 region of Brazil
Ribes De Lima, L 1
Universidade Federal de São Carlos, Brazil
Croton L., the second largest and the most diverse genus of Euphorbiaceae, has about 1,300 species, of herbs, shrubs, and trees that are ecologically prominent and often important elements of secondary vegetation, grouped in 40 sections. It has a pantropical distribution with the majority of species occurring in the Americas.
The genus is most diverse in semiarid regions, but also occurs in habitats ranging from beaches to tropical rain forests. In South America, most of the species –about
356– are in Brazil.
Of which, approximately 100 are found in Southeast region, most likely the center of diversity of the genus. Croton is a typical example of a
‘problem genus’, i.e., big and complex regarding taxonomy. That may be the reason genera that are smaller and better delimited are widely studied. Because of its size and diversity, no complete taxonomic treatment in relation to this genus was accomplished since Prodromus and, in Brazil, since Flora brasiliensis.
Despite the great morphological diversity, in its current delimitation, Croton is a monophyletic genus well characterized by terminal inflorescences, stamens curved in flower button, by diverse arrangements of the unisexual flowers and different kinds of specialized glands. Brazilian Southeast region is composed of four
States. Espírito Santo State total area is 46,184.1 km².
Moreover, its climate varies from dry tropical to highaltitude tropical climate; the former one occurs in the coast range. Minas Gerais State is 586,523 km² and tropical and high-altitude tropical climates are dominant.
The highest regions and the south of the State have the lowest temperatures, almost reaching 0°C. Rio de Janeiro
State total area is 43,696.054 km². Also, its types of climate are tropical in the lowest areas, and high-altitude
613

tropical in high altitude plateaus. São Paulo State is
248,256 km²; furthermore, the Tropic of Capricorn passes through it. The climate of this State is characterized by dry and wet seasons well defined in the majority of the State, except for the Serra do Mar, near the coast, where the dry season is very short. The biomes found in Brazilian Southeast region are dense ombrophyla forest, seasonal forests, and cerrado. The main purpose of this study is to make a floristic survey of the genus Croton in southeast region of Brazil through expeditions to collect specimens and visits to the main herbaria of this region, seeking greater knowledge and delimitation of species, and also the biodiversity of this area. This study will contribute to a better understanding in relation to taxonomic aspects and phylogeny of Croton globally. Besides, data regarding morphology, biology, biogeography, and habitats of the species are collected.
The final objective of this study is to elaborate an
Illustrated Guide to Identify Species of Croton
(Euphorbiaceae s.s.) of Brazilian Southeast Region, dividing species into groups, first by habit, then by type of indumentum, etc. It aims at making the use of the key easier for non-experts because, frequently, analytical identification keys use features difficult to understand, as a consequence, only experts can use them.
P0874 – ePoster
Insights into the infrageneric classification and biogeography of the genus
Athyrium
(Athyriaceae,
Pteridophyta) from chloroplast DNA sequence data
Liu, Y-C 1 , He, Z-R 2 , Zhang, X-C 3 , Chiou, W-L 4
1 Dept of Biological Resources, Chiayi University, Chiayi,
2 Taiwan; Institute of Ecology and Geobotany, Yunnan
University, Kunming, China; 3 State Key Laboratory of
Systematic and Evolutionary Botany, Institute of Botany,
Chinese Academy of Sciences, Beijing, China; 4 Division of Botanic Garden, Taiwan Forestry Research Institute,
Taipei, Taiwan
The lady ferns ( Athyrium Roth) comprising about 200 species, are mainly distributed in temperate and tropicalsubtropical mountainous regions of Asia. Nucleotide sequences of the chloroplast trn Ltrn F, rbc L, atp A, rps 4trn S, and mat K are obtained from over 70 species representative of the infrageneric classification.
Integrating those molecular data and morphological characteristics, seven clades were strongly supported and recognized as sections. Among them, species of sect.
Athyrium are mainly distributed in the temperate zone of the northern hemisphere; sect. Wallichiana and
Yokoscentia are restricted distributed in Himalayan and
NE Asia respectively. These sections are the basal clades in the Athyrium phylogeny tree. Section Biserrulata ,
Echinoathyrium , and Strigoathyrium are widely distributed in eastern Asia, somewhat extending to India and Africa. The members of sect. Polystichoides are endemic to Himalayan or disjunctively distributed in this area and on western Pacific islands. These biogeographical patterns are considered as the results of vicariance or/and long distance dispersal. The ratio of endemism in the western Pacific islands is high, especially in the tropical regions, which include many mountainous islands. They are at the margin of the Asian continent and experienced the repetitive glacial and
614 interglacial periods. Our results show that those historical geographic events and the margin effect have caused the formation of these endemics and their further evolution.
P0875 – ePoster
Molecular phylogeny of
Cissus
L. (Vitaceae), based on nuclear rDNA (ITS) and chloroplast (trnL-trnF) sequences
Rodrigues, JG 1
1
MB 1
, Lombardi, JA 2 , Novaes, RML 1 , Lovato,
Federal University of Minas Gerais, Belo Horizonte,
Minas Gerais, Brazil; 2 State University of Sao Paulo, Rio
Claro, Sao Paulo, Brazil
Cissus is the largest genus of Vitaceae with perhaps 350 species in the entire world. The chief diversity centers for the genus are South America and Tropical Africa.
Because Cissus is a large, heterogeneous taxon with delimitation problems we conducted phylogenetic analyses using two non-coding sequences of chloroplast
DNA ( trn L intron, and trn Ltrn F intergenic spacer region) and nuclear rDNA (ITS-1, 5.8S rRNA and ITS-
2). The sequences of 50 Cissus species, 27 of which grow in South America, 17 in other continents, and six outgroup species of other Vitaceae genera, were analyzed using maximum parsimony, maximum likelihood and
Bayesian inference. The analyses showed that Cissus , as presently defined, is not monophyletic. The South
American species Cissus tweedieana, C. simsiana and C. striata are inferred as closer to the Clematicissus clade than to any other South American Cissus species, results that are consistent with the morphological data. The
South American species Cissus trianae and the Australian species C. hypoglauca, C. oblon ga and C. antarctica formed a distinct clade, apart from any other species. The other analyzed species formed a consistent clade including most of the sampled taxa. The next steps include: 1) finding out in which clade Cissus vitiginea L., the type species of the genus, should be placed to determine the species that will bear the genus name; 2) include sequences from other outgroup species, and 3) making the necessary nomenclatural realignments.
P0877 – ePoster
Biogeography and systematics of Musaceae in
Indochina: molecular morphological approaches phylogenetic and
Ly, NS 1 , Haevermans, T 1 , Lowry II, PP 2
1 Muséum National d’Histoire Naturelle, Département
Systématique et Evolution, CNRS-Origine Structure et
Evolution de la Biodiversité, Paris, France;
Botanical Garden, St Louis, MO, USA
2 Missouri
Musaceae is a small family a majority of whose ca. 65 species in 3 genera occur in tropical Asia, Africa and
Australia. Members of the family are an important food crops, most notably edible bananas but also beverage products, and sources of fibres, medicines, ornamentals, etc. Cambodia, Laos and Vietnam, which lie within the
Indochinese megabiodiversity ‘hotspot’, are the family’s centre of diversity. The first account of bananas in

Indochina can be found in Loureiro’s Flora cochinchinensis (1790), where 5 species and 3 varieties are treated in a single genus, one of which was described there. Gagnepain’s revision for Indochina (1908) provided the first substantial regional account, with 1 genus and 6 species. Nearly 90 years later, a summary of
Musaceae was compiled by Ho P.H. in his Cay Co Viet
Nam (2000), with 3 genera (including Ravenala ) and 16 species, though some accepted species names had changed and synonymy had been revised. However, the family is far from adequately known in Indochina: many areas remain totally unexplored and many herbarium collections have not yet been studied. Field research in the region is therefore essential to understand the biology, morphology, phylogenetic relationships in this family, and thereby enable a better interpretation of its biogeography and evolutionary history. Field work is being focused on protected areas throughout Indochina, emphasizing places from which no herbarium material is available. Data gathered to date suggest that understanding Musa in Indochina may have been complicated in the past by the occurrence of hybridization among and within species and by polypoidy, which has led to inconsistent levels of morphological variability among the taxa historically recognized. To improve our understanding of the taxonomy, diversity, biogeography and evolution of
Musaceae in Indochina, research towards a revision and a robust phylogeny is now being conducted.
P0878 – ePoster
Some interesting plants from Mt Abu
Mahmoudi Otaghvari-Kelij, A-S 1
1 University of Mazandaran, Iran
Mt Abu (1727 m) at the south-western border of the
Rajasthan is the highest peak not only of the Aravallis but also between western Himalays and Nilgiri Hills. The vegetation here falls into fairly distinct elevational zones though they intermingle to certain extent. The chief components up to 1300 m In are the same as between
Bijapur and Mt Abu with only a little change in the frequency and abundance. But above 1300 m, the vegetation gradually changes to subtropical evergreen type with species like Boswellia serrata, Carvia callosa,
Crateva nurvala, Flacourtia indica, Girardinia zeylenica, Jasminum humile, Lannea coromandelica,
Mallotus philippensis, Mangifera indica, Rosa brunonea,
R. involucrate, Sterclia urens, Syzgium cumini, etc. At certain places Albizia spp., Erythrina spp., Emblica officinalis, Kydia calycina, Trema orientalis , etc., are also found at higher elevations. Several species of ferns and fern allies occur at Mt Abu.The present paper presents preliminary observation on plants of Mt Abu.
P0879 – ePoster
History of cultivated
Genista
, originating from
Macaronesia
1
Malécot, V 1
UMR GenHort, Angers, France
Macaronesian Genista , particularly from section Teline
(= genus Teline Medik.), have been the source of a few cultivated, and possibly now invasive, brooms. For example, millions plants of Genista x spachiana are sold in Europe under various names (including Cytisus racemosus ), while it is reported as invasive in California and Australia. This taxa, expected to be a hybrid between
Genista stenopetala and G. canariensis, occurs in the wild only in Tenerife. A taxonomic study involving both wild and cultivated material assigned to Genista x s pachiana shows that extant wild individuals may not be hybrids but are molecularly closely related to G. canariensis .
Extant cultivated samples of G.
x spachiana are of hybrid origin, involving G. stenopatala, G. canariensis but also
G. monspessulana . In addition, historically cultivated
Cytisus racemosus – a presumed synonym of Genista x spachiana – are identical to G. stenopetala . Using herbarium samples and horticultural reports we clarify this complex story. Denomination changes between 1830 and 1850 by Philipp Baker Webb are a first source of confusion. Exchanges between botanical gardens, commercialisation and propagation by seedlings around
1840 and 1870 are a second source of name proliferation.
To keep current usage, various changes are needed in this group.
P0880 – Poster
1
Evolution of the
Chenopodium album
group
Pastova, L 1 , Krak, K 1 , Mandak, B 1
Institute of Botany, Pruhonice, Czech Republic
For the study of Chenopodium album aggregate we have selected those species of Euroasiatic origin that represent the diploid–polyploid complex, i.e. C. album, C. ficifolium, C. opulifolium, C. pedunculare, C. striatiforme, C. strictum and C. suecicum . They give us a unique opportunity to study evolutionarily processes for several reasons: (a) the three ploidy levels (2x, 4x, 6x) occur within one territory in almost the same habitats, (b) within each ploidy level, several species have been recognised, (c) hybridisation occurs, but its frequency is unknown, (d) there is only limited knowledge concerning the relationship between morphology and ploidy levels as well as (e) the range of phenotypic plasticity of particular species with different ploidy levels. Using classical caryology, flow cytometry and in situ hybridization we have identified the origin of polyploid taxa within
Chenopodium album agg.
P0881 – ePoster
Studies on spore morphology and qualitative analysis of free amino acids of some pteridophytes with special reference to their ethno-medicinal uses in West
Bengal, India
, Mondal, AK 1 Mandal, A 1
1 Vidyasagar University, India
The diversity of Pteridophytes in West Bengal, India is very rich and always attracts this diversity to the pteridologists. Spore morphology is considered as an important tool for identification and characterization of
615

fern species. In the recent years the biochemical data also play an important role in characterization and determination of inter-specific relationship among the taxa . A prolonged survey has been done at the Gangetic delta of West Bengal, India and 29 species have already been identified and documented. Our study also reveals the diversification of spore morphology in between the identified species. The qualitative analysis of free amino acids was also analyzed by TLC. The identified amino acids are cystine, glutamic acid, tyrosine, serine, methionine, proline etc. The ethno-medicinal uses also reported in our present paper .
P0882 – ePoster
Diversity and evolution of extrafloral nectaries in
1
Sonoran desert plants
Marazzi, B 1
JL 1
, McMahon, MM 2 , Lanan, M 1 , Bronstein,
University of Arizona, Dept of Ecology Evolutionary
Biology, USA; 2 University of Arizona, Dept of Plant
Sciences, USA
Desert plants face great challenges to growth and survival from herbivorous insects. To cope, many plants utilize ‘biological warfare’: they reward aggressive ants with carbohydrate-rich, liquid secretions from extrafloral nectaries (EFNs), and the ants in turn attack the plants’ enemies. Such ant-plant mutualisms are well studied in tropical and savannah habitats, but are not well understood in desert regions. Yet desert EFN-based mutualisms are particularly intriguing, because it is not clear how plants can effectively manage their limited water budget to produce a water-based reward such as extrafloral nectar. Focusing on the northern Sonoran
Desert flora (USA), we (i) catalogued EFN-bearing plant species, (ii) investigated EFN morphology to identify patterns of convergent morphological adaptation to the arid climate, and (iii) reconstructed the evolutionary history of (selected) EFN-bearing plants using dated phylogenies to explore whether EFNs were present prior to the formation of the desert. (i) Our survey identified up to 78 genera from 32 angiosperm families that may include one or more species with EFNs in the Sonoran
Desert and adjacent higher elevation habitats.
Leguminosae dominated with 28 EFN-bearing genera
(36% of total EFN genera), followed by Euphorbiaceae
(up to 9 genera), Rosaceae (4 genera), Cactaceae and
Asteraceae (3 genera). For our morphological and evolutionary analyses, we focused on legumes, the dominant element in the Sonoran Desert and other kinds of vegetation worldwide. (ii) EFNs in most legumes are located on leaves, on the petiole and/or between pairs of leaflets, including phyllodes (i.e., highly modified and xerophytic leaves), and sometimes on stipules, stipels, and/or pedicels. Our anatomical studies of foliar EFNs in selected species of the legume genus Senna show that the secretory parenchyma is reduced in number of cell layers and tissue area compared with EFNs in non-desert species. (iii) Using a dated molecular phylogeny of
Senna , we inferred the evolutionary history of a clade of desert Senna EFN-species distributed across the North
American deserts (Sonoran, Chihuahuan and Mohave deserts). Because EFNs in Senna originated ca. 40 million years ago, before the formation of the North
616
American deserts, this desert EFN clade is derived from an ancestor that most likely already possessed EFNs and then diversified along with the aridification and origin of the deserts. EFNs have apparently been reduced but have been retained in a functional state (i.e., secreting nectar) in most desert legumes, and are thus capable of participating in protective mutualisms with desert ants.
P0883 – ePoster
Evolution of extrafloral nectary diversity in
Senna
Marazzi, B 1 , Bronstein, JL 1 , Sanderson, MJ 1
1 University of Arizona, Dept of Ecology Evolutionary
Biology, USA
Most key innovations in plants that have been studied to date are traits directly involved in one of three major plant–animal interactions: pollination, seed dispersal, and herbivore defense. Extrafloral nectaries (EFNs) have played a key role in the diversification of the widespread genus Senna (Leguminosae; 300-350 species). EFNs are secretory structures on plant parts outside the flower, offering nectar to ants in return for protection from herbivores. Senna ’s ‘EFN clade’, which includes about
80% of species within the genus, is characterized by conspicuous EFNs on the leaves and sometimes the pedicels. The morphological diversity of these EFNs is poorly known. Disentangling its evolution could provide new insights to increase our understanding of the role of
EFNs as key innovations in and beyond the
Leguminosae. Therefore, we here characterize the diversity of EFNs in Senna and reconstruct their evolution in a phylogenetic framework . EFNs can be found in eight different locations in Senna : on the leaves
1) at the base of the petiole, 2) between the first pair of leaflets only, 3) between the first and subsequent pairs of leaflets only, and 4) between all pairs of leaflets; and on the inflorescence 5) at the base of pedicels, 6) on the stipules, 7) on the bracts, and 8) on the calyx. Foliar and pedicellar EFNs are ’true‘ EFNs (i.e., true secretory organs). In contrast, stipular, bract and calyx EFNs, which are reported here for the first time, are so-called
‘substitutive’ EFNs (i.e., modified existing structures) and appear to occur only in species of clade II, previously considered a non-EFN clade. True EFNs can be sessile or subtended by a stalk; the shape of the secretory portion ranges from globose, ovoid, pyriform to cylindrical. Our anatomical studies focused on true EFNs
(from at least nine microtome-sectioned Senna species).
The typical structure of nectar-producing organs was identified. It consists of the epidermis, the nectary parenchyma, and the subnectary parenchyma. In addition, a fourth layer of lignified cells is found between the nectary and the subnectary parenchyma. The function of this layer is unclear, but it might serve as a support structure resulting in the convex shape of true EFNs in
Senna. Parsimony reconstructions of EFN location on the
Senna phylogeny suggest that foliar EFNs first evolved between the first pair of leaflets only. Later, additional
EFNs independently evolved in multiple clades on subsequent pairs of leaflets. EFNs at the base of the petiole are derived, having evolved in only a single clade.
Pedicellar EFNs appear to have evolved once in the common ancestor of clades V and VI. Substitutive EFNs might represent an unrecognized synapomorphy of clade

II. Finally, true EFNs seem to possess a distinctive evolutionary trajectory, whereas substitutive EFNs appear integrated as part of the evolutionary trajectory of their subtending organ (stipules, bract, calyx). We discuss these differences in morphological evolution of true vs. substitutive EFNs in Senna in the context of the evolutionary role of EFNs as a key innovation and their ecological role in ant-plant mutualisms.
P0884 – ePoster
How many distributional patterns are found in
Japanese
Carex
(Cyperaceae)?
Masaki, T 1 , Hoshino, T 1
1 Okayama University of Science, Japan
Cyperaceae is one of the largest family in the world, ca.
102 genera and 5000 species were recorded
(Goetghebeur 1998). In Japan, plants of the Cyperaceae are found growing everywhere, from the seashore to alpine regions. They are especially common in wetlands along lakes and rivers. Twenty-six genera and about 493 taxa were reported in Japan (Hoshino & Masaki 2011).
Over 300 taxa have been recorded in the Japanese genus
Carex , the largest genus of flowering plants in Japan.
About half of the taxa of the genus Carex are endemic to
Japan. Distribution maps of Japanese Cyperaceae based on herbarium specimens have been reported (Kanai et al
2008). Our study aims to clarify distributional patterns of all Japanese Carex in the Japanese Archipelago, based on enormous specimens data. We used totally 14327 specimens deposited in Herbarium of Hokkaido
University Museum (SAPS), National Museum of Nature and Science, Tokyo (TNS), Kanagawa Prefectural
Museum of Natural History (KPM), Natural History
Museum and Institute, Chiba (CBM), and other 13 facilities. We used mainly three kinds of free application software, Kenmap by chirashi-zushi for plotting dots,
Uocchizu by Geospatial Information Authority of Japan for identifying location, and Paint.NET by dotPDN LLC et al. for image processing. In addition, we made an
Excel file for data conversion, it can convert Global
Positioning System data to csv format data for Kenmap.
After data converted, open Kenmap software and next load the csv format data. Kenmap have blank Japanese maps which include all 47 Japanese Prefectures and detailed cartographic information. A distribution map is saved as png format data. Finally, each species name and other information are added in maps using layers. We have already finished making distribution maps of 84 taxa. Our results show following eight distribution patterns; 1) Hokkaido, 2) disjunct distribution in
Hokkaido and Central Honshu, 3) from Hokkaido to
Central Honshu, 4) Hokkaido, Honshu, and Kyushu, 5)
Honshu, 6) Honshu, Shikoku, and Kyushu, 7) from
Honshu to Ryukyu, and 8) found in only one or two places. We found the pattern 3 is most common, and pattern 7 is few.
P0885 – ePoster
Miocene fossil forest from Shimokawa, Hokkaido,
Japan
Matsumoto, M 1 , Kashiwabara, K 2
Terada, K 3
, Yasukuni, Y 1 ,
1 Faculty of Science, Chiba University, Japan;
2 Shimosato High School, Japan;
Dinosaur Museum, Japan
3 Fukui Prefectural
The middle Miocene Mosanru Formation (ca.12 Ma),
Shimokawa, northern Hokkaido, Japan contains fossil forest (upright stumps), petrified wood (both drifted and transfer), permineralized plants and other plant macrofossils. The Mosanru Formation is composed of volcanic and sedimentary rocks and is subdivided into lower and upper parts by their lithologic and sedimentary facies. The lower part is mainly exposed in the Nayoro
River, eastern part of Shimokawa, and contains basal conglomerates, volcanic tuff and sedimentary rocks representing as terrestrial, cyclic fluvial channel flow.
These sedimentary facies are represented as flood plain and fluvial river side environments. The in situ stumps grew in unstable environments characterized by fining upward sequence with seventh cyclic-interval floods. The fossil assemblage in the second and third cycles, upright stumps and petrified woods indicate deciduous broad leaf community: Salix, Alnus, Ostrya, Cercidiphyllum and
Acer . These plants represent a fluvial ravine and river side community. In the forth cycle, petrified wood assemblage: Salix , Ostrya and Cercidiphyllum . In the fifth cycle, in situ stumps and roots assemblage: only
Picea . By the environmental aspect, this genus was adapted and confined to wetland. In the sixth cycle in situ stumps: Ostrya and Cercidiphyllum . The upper part of this formation is exposed in and around the Nayoro
River, the Mosanru River and the Rubeno-sawa, in the central part of Shimokawa. This part has fining upward sequence that terminated with seven peaty silicified layers (S1~S7). Near the S1 layer, conglomerates deposits contain with petrified woods of conifer and deciduous broad leaf plants: Abies , Picea , Pseudotsuga ,
Taxodioxylon , Pterocarya , Populus and Cercidiphyllum .
These petrified wood assemblage represents mixed with upland, river side and wet land taxa, characterized by slightly unstable environment. Other than the stumps and petrified woods, anatomically well preserved permineralized plants were reported from the silicified layers (S1–7) in the upper part of the Mosanru
Formation. The thick dark-black quartz beds each associated with lignite bed below. Abundant remains of several organs of Glyptostrobus rubenosawaensis
(Matsumoto et al., 1997) in the S3 layer, indicate that the petrified wood of Taxodioxylon cumminghamioides is highly assumed as Glyptostrobus , because other cupressoid and taxodioid taxa are not found.
P0886 – ePoster
Modeling gene flow in a lineage of Mexican red oaks: a case study of
Quercus
section
Lobatae
subsection
Racemiflorae
McCauley, R 1, 2 , Cortés-Palomec, A 1 , Oyama, K 1
1 Centro de Investigaciones en Ecosistemas, Universidad
Nacional Autónoma de México, Morelia, Michoacán,
México; 2 Fort Lewis College, Durango, Colorado, USA
617

The genus Quercus L. is well known for frequent hybridization among closely related species and recurrent hybridization is an important evolutionary process in the generation of biodiversity in this globally important group of forest trees . Many studies have shown that in
Quercus geography often plays a more important role in determining population genetic structure than species membership due to the prevalence of gene sharing among taxa . But morphologically discrete species are still maintained even in the light of high levels of gene exchange and sympatry . Are these patterns of morphological species maintained over long periods of time in these areas of sympatry or is there directionality in gene flow toward select taxa? To address this question we modeled gene flow within four closely related species of Quercus Section Lobatae Loudon Subsection
Racemiflorae A. Camus in the Sierra Madre Occidental of Mexico. Mexico is one of the centers of species diversification of Quercus with approximately 160 species occurring within the country . It is also the center of diversification of the Section Lobatae , the Red Oaks.
The Subsection Racemiflorae form a natural group of four species ( Q. conzattii Trel., Q. radiata Trel., Q. tarahumara Spellenb., J.R.Bacon & Breedlove, and Q. urbanii Trel.) extending across the Sierra Madre
Occidental and Southern Cordillera and are united by their production of fruits on long racemose inflorescences . The group has a wide but patchy distribution restricted to small habitat 'islands' due to an affinity for exposed mineralized soils. Most notable is a bicentric distribution, dividing the group by more than
700 km. Two species of Racemiflorae span this disjunction and show only minor morphological differences across their range. Using 49 populations and a combination of chloroplast and nuclear mirosatellites we have shown that the group exhibits a mosaic of chloroplast haplotypes representing long term hybridization and a pattern of nuclear genetic variation closely tied to geographic distribution . While across most of the range the species are generally isolated from other members of the same Subsection, the four taxa occur in close sympatry in the Pine-Oak Woodlands of Durango and adjacent Sinaloa. Closer examination of this area of sympatry using nuclear markers and an estimation of migration rate illustrates the pattern of gene flow among the individual taxa and shows a dynamic pattern of gene exchange among the predominant and less common taxa.
P0887 – ePoster
The early angiosperm record in Australia
McLoughlin, S
3
1 , Cantrill, D 2 , Wagstaff, B 3 , Tosolini, A-
MP
1 Swedish Museum of Natural History, Stockholm,
Sweden; 2 National Herbarium of Victoria, Royal Botanic
Gardens, Melbourne, Australia; 3 School of Earth
Sciences, University of Melbourne, Australia
Cretaceous angiosperms have been recorded in Australia since the early Twentieth Century. However, no comprehensive compendium of angiosperm fossils or analysis of their affinities has yet been undertaken. The angiosperm record on this continent is derived from three principle sources: 1, sparse macrofossils (mostly leaf and flower impressions); 2, dispersed cuticle and seeds
618 recovered from mesofossil residues; and 3, extensive pollen records that provide a moderately robust dataset for evaluating the timing of major taxon appearances.
The Early Cretaceous macrofossil record derives primarily from rift basins along the southeastern
Australian margin (Otway and Gippsland Basins) and central eastern Australia (Maryborough and Styx Basins).
These basins host up to 15 Early Cretaceous angiosperm leaf morphotypes. The oldest Australian angiosperm records, from the Aptian Koonwarra fossil bed of the
Gippsland Basin, consist of palmate- to pinnate-veined leaves and inflorescences attached to a slender stem, and are interpreted to possess a mosaic of characters shared by the Magnoliidae and basal monocots. Younger
(Albian) assemblages of eastern Australia are notably richer in angiosperm leaves and contain mostly simple lorate to ovate, entire or weakly toothed forms. Late
Cretaceous angiosperm macrofossils are represented only in Cenomanian deposits of the broad epicratonic
Eromanga Basin of central eastern Australia, although extensive untapped assemblages of Turonian –
Maastrichtian dispersed cuticles probably exist in rift basins along the southern continental margin. At least 12 leaf and flower types have been documented from the
Cenomanian, although the dispersed pollen record from these strata suggests that true angiosperm diversity was several times this number by the beginning of the Late
Cretaceous. Most of the leaf fossils from this time are simple toothed or lobed leaves with putative lauralean, fagalean, chloranthalean or aquifolialean affinities. By the Cenomanian, angiosperms co-dominated macrofossil assemblages with conifers in terms of taxon diversity and leaf abundance, and this record is matched by evidence of a dramatic rise in angiosperm pollen diversity and abundance from the middle Albian to Cenomanian. The mid-Cretaceous radiation of angiosperms also corresponds to a steady decline in the abundance and diversity of several gymnosperm groups with long
Mesozoic records (e.g. Corystospermales, Pentoxylales,
Bennettitales, Ginkgoales), although several of these groups are represented by late-surviving relictual taxa in the early to mid-Cenozoic of southeastern Australia. The mid-Cretaceous rise to dominance of angiosperms is matched in other parts of the world. The Australian fossil record shows that this continent was part of a truly global transformation of the vegetation. Post-Cenomanian angiosperm macrofossil assemblages have not yet been documented from Australia, and this remains one of the major gaps in palaeobotanical understanding of the evolution of the Australian flora, since it likely incorporates the differentiation of many of the continent’s iconic extant plant taxa.
P0888 – ePoster
Lord Howe Island
Spiridens
– evidence for an island endemic
Meagher, D 1
1 School of Botany, University of Melbourne, Australia
Since the 1930s, Spiridens muelleri Hampe from Lord
Howe Island has been considered a synonym of S. vieillardii Schimp. from New Caledonia, but on weak morphological grounds. To test the validity of the synonymy I obtained sequences for one nuclear and two

chloroplast DNA markers from S. vieillardii (Lord
Howe), S. vieillardii (New Caledonia) and three other species from the region, S. camusii, S. longifolius and S. reinwardtii . I also studied morphological characters to search for any significant differences. I found that there were significant genetic differences between the Lord
Howe Spiridens and the other Spiridens , as well as very distinct morphological differences not previously recognised. The Lord Howe Island Spiridens is clearly distinct from S. vieillardii , and must revert to its original name, S. muelleri. S. muelleri is therefore endemic to
Lord Howe Island and S. vieillardii is endemic to New
Caledonia .
P0889 – ePoster
Comparative patterns of genetic variation among populations of the
Zamia pumila
L. complex across three islands of the Greater Antilles
Meerow, A 1 , Francisco-Ortega, J
A 4 , Jiménez Rodríguez, F 4
Griffith, P 3
2 , Calonje, M
, Oberli, A 5
3 , Veloz,
, Stevenson, D 6 ,
1 USDA-ARS-SHRS National Germplasm Repository,
Miami, USA; 2 Florida International University and
3
Fairchild Tropical Botanic Garden, Miami, USA;
Montgomery Botanical Center, Miami, USA;
Domingo, Dominican Republic;
Centre, Kingston, Jamaica; 6
4 Jardín
Botánico Nacional D. Rafael Ma. Moscoso de Santo
5 Plant Conservation
New York Botanical
Garden, Bronx,, USA
The Zamia pumila L. complex (Cycadales: Zamiaceae) is a distinctive, monophyletic, diploid (2n =16) assemblage of populations restricted to the West Indies and southeastern U. S. (Florida) that is currently considered to encompass either a single polymorphic, or nine distinct species. We are extensively sampling populations of the group throughout its range and genotyping them with both microsatellite DNA (simple sequence repeat,
SSR) and sequences of single copy nuclear genes . To our knowledge, this will be the first systematic study of tropical plants contrasting sequence-based phylogeographic with SSR population genetic analyses.
Here, we present our analysis of sixteen SSR loci across three of the larger islands of the Greater Antilles:
Dominican Republic, Jamaica and Puerto Rico . Of the three islands, Puerto Rico has the greatest morphological diversity; three species have been recognized as occurring there by some accounts . Across 16 SSR loci that could be amplified across all three islands, Puerto
Rico has the highest allelic diversity in terms of expected heterozygosity and mean number of alleles per locus.
While Zamia populations from both the Dominican
Republic and Jamaica have similar gene diversities, only a single private allele characterizes the Dominican
Republic populations . Some populations in Jamaica show indications of genetic erosion, congruent with evidence of poaching and small population size. Mean pairwise Fst between populations is lowest in the Dominican
Republic, which we attribute to high levels of gene flow among the populations . In Puerto Rico, while Dest values among three populations each of Z. portoricensis and Z. pumila are 0.026 and 0.037, respectively, the mean among four populations of Z. erosa = 0.186, a 6-8 fold higher level of inter-populational differentiation. Genetic distance resolves all of the Jamaican populations nested within Z. erosa from Puerto Rico, which may suggest
Pre-colombian human inter-island movement of plants, since these two islands have never been connected by land, and Zamia has limited long-distance dispersal capability . Results of this project will be interpreted within the context of the complex environmental history of the region. We anticipate that an understanding of the genetic structure of the populations of this complex will help to delineate future conservation strategies, and serve as a model for population level studies of other rare
Caribbean plant taxa.
P0890 – ePoster
Expanded nrDNA ITS/5.8S phylogeny for
Amaryllidaceae tribe Hippeastreae (Asparagales)
Garcia, N 1 , Meerow, AW 2 , Soltis, PS 3 , Soltis, DE 1
1
2
University of Florida, Dept Biology, Gainesville, USA;
USDA-ARS-SHRS National Germplasm Repository,
Miami, USA; 3 Florida State Museum of Natural History,
Gainesville, USA
The Hippeastreae represent a subclade within the
American clade of the cosmopolitan Amaryllidaceae s.s. and contain several species relevant for horticulture, such as Hippeastrum spp., Habranthus spp., Zephyranthes spp., Rhodophiala bifida , and Sprekelia formosissima .
Despite the long-standing interest in these plants, their taxonomy at the generic level has been controversial, with several segregates proposed during the last 40 years.
A previous phylogenetic analysis of the nrDNA ITS/5.8S region showed that certain genera are not monophyletic, such as Rhodophiala , Habranthus , and Zephyranthes , but lacked good representation of Chilean–Argentinean groups, such as Famatina , Phycella, Placea,
Rhodolirium , and Traubia . The hypothesis of possible early lineage reticulation in the group was suggested. We have expanded the taxon sampling for this data set by including members of the Chilean endemic genera and additional species of Habranthus , Rhodophiala,
Rhodolirium, Sprekelia , and Zephyranthes . A total of approximately 110 species correponds to more than 60% of the tribe's species-level diversity. The tribe comprises two major clades: a) Traubia, Placea, Phycella,
Rhodolirium , and Famatina maulensis , characterized by x = 8, lack of polyploidy, and a capitate stigma, and b)
Rhodophiala, Habranthus, Hippeastrum, Sprekelia ,
Zephyranthes , and the remainder of Famatina , characterized by several basic chromosome numbers ranging between x = 6 – 11, and frequent polyploidy and aneuploidy. No clear morphological features diagnose the latter clade. The first group appears to be composed of three main subclades: 1) Rhodolirium laetum and
Traubia modesta , 2) Rhodolirium montanum , and 3)
Phycella spp., Placea spp., Rhodolirium speciosum , and
Famatina maulensis . The other major clade appears to be composed of four lineages with unresolved relationships:
1) Rhodophiala bifida , 2) Hippeastrum spp., 3) Eithea blumenavia , Chilean–Andean Rhodophiala spp., remaining Famatina spp., and South American–
Caribbean–southeastern US Zephyranthes spp., and 4)
South American–Mexican–southwestern US–Texan
Zephyranthes spp., Habranthus spp., Sprekelia spp., and
Haylockia americana . Certain species within
619

Hippeastrum , Rhodophiala, Placea , and Phycella have identical or almost identical sequences to others, implying a sort of interspecific nrDNA concerted evolution and/or perhaps rapid radiations within these groups. We are currently working with low-copy nuclear genes and several chloroplast regions to test further this phylogenetic hypothesis; therefore the ITS/5.8S topology is only a first step towards an integrated phylogeny of the clade. This framework will serve as a basis for a phylogenetic classification of the group, study of its chromosomal evolution, and a test of the ancient interclade reticulation hypothesis in Hippeastreae.
P0891 – ePoster
Phylogenetic relationships within the tribe
Polygonatae (Ruscaceae) and their biogeographic evolution in the Northern Hemisphere
3 Meng, Y 1,2 , Nie, Z 1 , Wen, J 3 , Yang, Y
1 Key Laboratory of Biodiversity and Biogeography,
Kunming Institute of Botany, Chinese Academy of
Sciences, Kunming, PR. China;
Plateau Research at Kunming, CAS, Kunming, PR.
China; 3
2 Institute of Tibetan
Dept of Botany, National Museum of Natural
History, Smithsonian Institution, Washington, DC, USA
The tribe Polygonatae (including four genera:
Maianthemum , Polygonatum , Heteropolygonatum , and
Disporopsis ) is well supported to be monophyletic group by previous phylogenetic studies, but phylogenetic relationships within the tribe and its biogeographic patterns within the Northern Hemisphere are still poorly understood. Extensive samplings through the whole tribe and its distribution range are badly needed to solve these issues. The monophyly of the tribe and each genus is well supported based on our phylogenetic analysis using four chloroplast and nuclear markers ( trn L-F, psb Atrn H, rbc L, trn K, and ITS) with a total length of over 5,000 base pairs. Disporopsis is first split from the other taxa of the tribe and then Maianthemum is sister to a clade including Polygonatum and Heteropolygonatum . Almost all species of Maianthemum from the eastern Himalayan region in SW China form a well supported clade.
Maianthemum tatsienense and M. stenolobum from SW to central China form another clade. The other
Maianthemum species from eastern Asia (central to NE
China and Japan) and the New World fall into several clades. For Polygonatum , two clades are strongly separated: one includes all species with verticillate or opposite leaves and the other is composed of all taxa having alternate leaves. The intercontinental disjunctions between eastern Asia and North America are found in both Polygonatum and Maianthemum . Their divergence times are estimated to be recent in the late Miocene to the
Pliocene with the Bayesian relaxed dating. Long distance dispersal by birds may have facilitated the evolution of their intercontinental disjunction and their biogeographic diversifications in SW China. A recent radiation in the
Pliocene is suggested in the high mountains of SW
China, corresponding to the geographical heterogeneity in that region after the uplift of the Himalayas.
620
P0892 – Poster
Metabolomic profiling: a promising new molecular approach in plant taxonomy and systematics
Messina, A
Walsh, NG 3
1 , Green, PT 1 , Hoebee, SE 1 , Callahan, DL 2 ,
2
1 La Trobe University, Bundoora, Australia;
Metabolomics Australia, School of Botany, University of
Melbourne, Melbourne, Australia; of Victoria, South Yarra, Australia
3 National Herbarium
Molecular methods have long been used as a companion to traditional morphological approaches in plant taxonomy and systematics, but the field is evolving. For decades the mainstay was flavonoid diversity determined by spot patterns in two dimensional paper chromatography, a popular method because flavonoids are simple to extract, cheap and quick to analyze, and with universal application across plant groups . However,
DNA sequencing has largely replaced chemical analysis because unlike flavonoid composition, genetic characters are not influenced by environmental factors, and have the added bonus of revealing phylogenetic relationships .
These advantages, together with the development of inexpensive and widely applicable protocols, have seen
DNA sequencing become a standard method in plant taxonomic and systematic studies, particularly for higherorder analyses. However in many taxonomic studies,
DNA sequence data does not provide sufficient resolution to delimit closely related species, and unlike the COI region in animals, a standard region for species separation of plants has not been found. Other molecular methods such as microsatellite and low copy nuclear genes have been developed to detect lower level variation for some plant groups, but the markers are not universal and development of new markers can be time consuming and expensive. Potentially, metabolite profiling offers a new approach to delimiting closely related species .
Rather than focusing on a single group of secondary metabolites such as flavonoids, mass spectrometry (MS) can now be used to generate full metabolite profiles .
These analyses are fully automated and rapid, producing profiles that include a vast array of compounds linked to gene expression. This approach has recently been used to delimit species and races of bacteria, and is now being developed as a tool for microbial species identification .
In this study, full metabolite profiles are being used to delimit taxonomic boundaries of several closely related plant species in the Olearia phlogopappa (Asteraceae) complex . A morphological study of the group has found several taxa, but chloroplast and nuclear DNA sequence data have been unable to resolve the group. Metabolite profiles are being used as an alternate molecular approach. Polarized compounds were extracted from leaf material in 80% methanol; profiles were then generated using MS linked with Liquid Chromatography. Profiles of 70 samples from 11 species found over 17 000 compounds. Many of these compounds were found to be unique to single specimens, so only those occurring in at least two specimens were included in analyses to remove uninformative data, for a final dataset of just over 10000 compounds. Multivariate analyses of this data support current species and subspecies concepts based on the morphological study of the group. Profiling also identified some potential subspecific taxa not fully

resolved by the morphological study . Metabolite profiling is a rapid and relatively inexpensive method that can be used across all plant groups, and in this study has been shown to have a great application in low-level plant taxonomy.
P0893 – ePoster
Climate change impacts on ferns: reconstructing the past to predict the future
Metzgar, JS 1 , Ickert-Bond, SM 1
1 Musuem of The North, University of Alaska Fairbanks,
Fairbanks, USA
Understanding the genetic consequences of changing species distributions is critical for evaluating the potential impact of climate induced distributional changes . The parsley ferns ( Cryptogramma ) make an excellent candidate for testing hypotheses of colonization following the Last Glacial Maximum (LGM). Thanks to their minute and easily dispersed spores, these rockloving ferns could have utilized refugia to rapidly and repeatedly colonize deglaciated landscapes, or they could have survived in situ on nunataks . Herbarium specimens indicate that Cryptogramma survives today on nunataks, but cannot answer whether this was the sole recolonization source used following the LGM or if recolonization from refugia also played a complementary or dominant role. By first generating well-resolved and robustly supported hypotheses of species relationships of the genus, we place the North American taxa in a phylogenetic context and answer lingering questions regarding (1) the origin of allopolyploid taxa and (2) if these taxa were formed when previous climate conditions forced currently allopatric parental taxa into close proximity . Field work throughout northwestern North
America sampled numerous populations and haplotype network analyses of these specimens reveal the contributions and locations of glacial refugia for
Cryptogramma during the LGM . These results also demonstrate the speed with which parsley ferns colonized deglaciated land following the LGM, which has considerable relevance for conservation efforts assessing future range shifts necessitated by anthropogenic climate change . Ecological niche modeling will allow us to project these future range shifts under a variety of climate scenarios and determine if polyploid species possess a lower extinction probability than diploid species.
P0894 – ePoster
Botany at the last frontier: the University of Alaska
Museum of the North Herbarium (ALA) showcasing digital resources and collection-based research to study the Arctic flora
Metzgar, JS 1,2 , Jeffers, SS 2 , Ickert-Bond, SM
1 Museum of The North, USA; 2
2
University of Alaska
Fairbanks, USA
The nucleus of the Herbarium at the University of Alaska
Museum of the North (ALA) had its origins in the 1920s as small, personal herbaria found their way to the
University of Alaska Fairbanks . In 1969 when David F.
Murray arrived to become Curator, the Herbarium consisted of about 40,000 vascular plant specimens and several shoeboxes of various mosses and lichens . The growth of the herbarium then exploded in the 1970s and
1980s, fueled largely by federal activities such as the establishment of several National Parks in Alaska, implementation of the Endangered Species Act, sponsored fieldwork in Alaska and abroad and exchanges and collaborations with other herbaria, especially focusing on the Russian Far East and central Siberia .
ALA currently houses approximately 230,000 specimens, including 130 type specimens and the collection is dominated by vascular plant (170,000 specimens), bryophytes (38,000) and lichens (17,000) . Comprising the largest collection of Alaskan plants in the world,
ALA also possesses significant holdings of Russian Far
Eastern and Canadian specimens . We have also recently added the J.P. Anderson collection from Iowa State
University . This valuable historical collection of 32,000 specimens forms the basis of Anderson’s seminal Flora of Alaska and contains early baseline specimens ranging from the 1890s to 1930s . ALA contains key information critical to questions of past and present biodiversity, and key indicators of local and global environmental change and sustainability . Studies of biodiversity, evolutionary history, ecology, and the management of the flora are enriched by specimens archived here . Specimen data for the vascular plants are available online through the public database Arctos (http://arctos.database.museum), with over 160,000 specimens also having high-resolution digital images available . This user-friendly database can be queried by taxonomy, locality, media attributes and by selecting a geographic area on a Google map . The herbarium is also actively involved in a number of current research projects using a combination of field work, morphological study and molecular analyses to resolve lingering taxonomic and evolutionary questions, especially with regards to the Arctic Flora . Our staff includes specialists focusing on revisionary work in
Ephedra, Altingiaceae, Oxytropis, Carex, Potentilla,
Eritrichium, Cryptogramma, Claytonia and the flora of
Kodiak Island . ALA staff is also active in floristic initiatives such as the Flora of North America project, the
Conservation of Arctic Flora and Fauna (CAFF) and basic inventory and monitoring programs supported by the federal government. ALA is crucial to the scientific and cultural infrastructure of the unique and important biogeographic region of Beringia.
P0895 – ePoster
A phylogenetic analysis of the tribe Miconieae
(Melastomataceae) based on DNA sequence data from nuclear and plastid loci
Michelangeli, F
R 4 , Becquer-Granados, E 5
Penneys, D 2
1 , Almeda, F
, Skean, JD 6
2 , Judd, W 3
, Ionta, G 3
, Nicolas, A 1
, Goldenberg,
, Ocampo, G 2 ,
1
4
New York Botanical Garden, USA; 2 of Sciences, USA; 3
California Academy
University of Florida, USA;
Universidade Federal do Parana, Brazil;
Botanico Nacional, La Habana, Cuba; 6
5 Jardin
Albion College,
USA
621

The tribe Miconieae (Melastomataceae) is the most diverse group in the family with over 2000 species in 19 genera . The tribe is characterized by flowers with partly to fully inferior ovaries that develop into baccate fruits, terminal or axillary multiflowered inflorescences (but not caulinar), and anthers without pedoconectives or well developed connective appendages . The largest genera in the tribe are Miconia (1100 species), Leandra (250),
Clidemia (230) and Ossaea (95) . However, generic delimitations have been traditionally complicated and several authors have in the past suggested that most of these are arbitrary and should be reworked entirely. Here we present a phylogenetic analysis of the tribe based four loci, two nuclear (nrITS, nrETS) and two plastid spacers
(psbKL and accD-psaI) . In total we sampled almost 800 species of Miconieae and over 50 species of outgroup taxa . In parsimony and maximum likelihood analyses the genera Eriocnema and Physeterostemon , both with capsular fruits and found in eastern Brazil, are resolved as sister to the Miconieae . Miconia is resolved as paraphyletic, with all other genera nested within it.
Leandra and Ossaea , both genera with acute petals, are highly polyphyletic, although the majority of species of
Leandra do form a large clade restricted to Eastern
Brazil.
Of the genera with over 30 species, the only one resolved as monophyletic is Mecranium , a genus endemic to the Greater Antilles. Other genera are recovered as monophyletic ( Anaectocalyx, Charianthus,
Killipia, Maieta , and Pleiochiton ), but none of these have more than 10 species, and they are all embedded within clades with species currently assigned to other genera. In spite of all of this taxonomic disagreement, these results do correlate well with biogeography and some morphological characters, particularly seed and anther morphology, and to a lesser degree inflorescence position . Other characters traditionally used as to define genera such as petal and hypanthium shape, have either been poorly implemented or are highly homoplasious .
P0897 – ePoster
1
Genetic variation and phylogeny of
Pandanus
,
Pandanaceae, in Japan
Miyamoto, J 1 , Susanti, R 2
Graduate School of Science and Engineering,
Kagoshima University, Japan; 2 Research Center for
Biology, Indonesian Institute of Sciences, Indonesia
The genetic variation and phylogeny of the Japanese species of the genus Pandanus L., Pandanaceae, were studied by comparing related taxa from Indonesia and
Micronesia . The genus Pandanus includes 600–700 species and distributes in the old world tropics . Pandanus odoratissimus is a beach species, broadly distributed in tropical area of India, Malesia, West Australia and its northern most distribution of Ryukyu Islands of Japan .
This species is sometimes confused with P. tectorius
Parkinson, another beach species distributed in the
Pacific area . Pandanus boninensis is an endemic species of Ogasawara (Bonin) and Iwojima Islands. This species can be found from seacoast to the hill ridge. Pandanus daitoensis Susanti et J.Miyam. was newly described from
Kitadaito of Daito Islands of Okinawa prefecture, Japan, in 2009 . This species is closest to P. duriocarpus Martelli morphologically, but can be distinguished by its two
622 rows of flexed acute carpels resulting in a surface of curved phalanges. Each carpel has dark brown longitudinal grooves extending from the stigma to the carpel base . Three hundreds and forty three plants were collected from 11 islands of Ryukyu, two islands of
Daito area and two islands of Ogasawara area and profiled using random amplified polymorphic DNAs
(RAPD) and inter simple sequence repeats (ISSR) analysis . Nucleotide sequences of four regions of cpDNA, rbc L, trn Htrn K, trn Strn T, psb Ctrn S, and three regions of mtDNA were analyzed . One accession from each island was used for the phylogenetic analysis .
Two or more accessions were used if two or more genetic groups were found within an island . As comparative taxa,
P. motoleyanus Solsm, P. helicopus Kurz ex Miq., P. dyctiotus St John, P. furcatus Roxb., P. nitidus (Miq.)
Kurz and P. odoratissimus from Indonesia, P. tectorius from Micronesia and Freycinetia formosana Hemsl.,
Pandanaceae, from Ogasawara Islands were used . As a result, the genetic variation of P. odoratissimus tended to be complicated. There was no clear partitioning of population of P. odoratissimus according to geographical area in Japan. The beach species were dispersed over long distance by sea current and had possibility of making hybrids with each other . The populations of P. odoratissimus in the Ryukyu and Daito Islands seemed to be influenced genetically from related taxa of the Pacific islands and the Southeast Asia. Pandanus daitoensis and
P. tectorius and P. odoratissimus were closely related to each other . Pandanus boninensis seemed to be an isolated endemic taxon to the Ogasawara Islands since it was not closely related to any taxa examined in this study.
P0899 – ePoster
Circumscription of the families within Leguminales as determined by cladistic analysis based on seed protein data
Mondal, AK
1
1 , Mondal (Parui), S 2
Vidyasagr University, Dept of Botany & Forestry, Plant
Taxonomy, Biosystematics and Molecular Taxonomy
Laboratory, Midnapore, West Bengal, India; 2 Lady
Brabourne College, Dept of Zoology (Section-
Biochemistry),Kolkata, West Bengal, India
Comparative banding pattern of the seed protein of 17 taxa belonging to the three families Mimosaceae,
Caesalpiniaceae and Fabaceae was studied with the aim of testing the proposed delimitation of the order
Leguminales (Fabales) into the three families or subfamilies and to assess the phylogenetic relationships within the three families. Cluster and pairing affinity or similarity index analysis of the data from total protein grouped the 17 taxa into three discrete clusters based on their families. Considerable amount of homology was observed in the banding pattern between the different taxa .
P0901 – ePoster
Botanical surveys in the Caatinga vegetation of northeastern Brazil: current situation and perspectives for studies of a poorly known seasonally dry tropical forest

1
Moro, MF 1 , Martins, FR 1
State University of Campinas, Campinas, Brazil
Caatinga is the main vegetation type in the semi-arid
Brazilian Northeastern Region, but it is one of the less known phytogeographical domains of the country.
Recently, floristic links between the Caatinga and other dry forests in the American continent have been stressed and some authors proposed that many disjunct dry forests in the continent may be treated as a larger phytogeographical domain called Seasonally Dry
Tropical Forests. To understand the distributional pattern of communities and know the floristic composition of
Caatinga, however, we need to perform a meta-analysis of the domain. Such analysis was undertaken for the
Atlantic Forest and Cerrado domains, but little information is available about Caatinga. The aim of this work is to compile floristic and phytosociological articles in order to assess how many surveys on this topic are available. We also intend to identify what phytosociological methods are applied more commonly and the mean richness of Caatinga surveys. We traced botanical papers in the main biological and botanical
Brazilian journals and in the journals indexed in the Web of Science database. We were able to gather 47 surveys published in 38 papers.
of this amount, 27 were floristic and 30 phytosociological. The most common method for phytosociological studies was the plot method, employed in 23 (77%) surveys, while the quarter method was used in 7 studies. Although Brazilian phytosociological survey protocols suggest at least 1 hectare in plot or 250 points in quarter method studies, 77% of the compiled phytosociological works sampled less than this recommendation (mean 0,63ha for plot studies and 130 points for quarters). The most common inclusion criteria used in Caatinga is to measure all plants with at least 3 cm of diameter at ground level (used in 60% of the surveys). The best studied Brazilian State was
Pernambuco (20 surveys), followed by Ceará (10).
Bahia state, the biggest in Northeastern Brazil, had just 2 surveys and no paper was found dealing with Caatinga vegetation in Alagoas and Sergipe states. The mean richness of woody species in the phytosociological works were 35 species and the mean richness in floristic surveys (many included all life forms, and some only the herbaceous or woody component) were 98 species.
Recent studies show that therophytes are the most common Raunkiaer’s life form in the semi-arid environments of Caatinga, and this component of the vegetation is not sampled by phytosociological work concerned with woody species. Four surveys were concerned exclusively with the herbaceous component, and these works had a mean richness of 70 non-woody species. Up to date, the Caatinga domain is very poorly represented by botanical inventories and currently it is important to increase the sampling effort, especially in the low studied Caatingas of Bahia, Sergipe and Alagoas.
Surveys concerned with the non-woody component are also very desirable, once the species richness is mainly concentrated in the herbaceous layer, not in the woody component.
P0902 – ePoster
Endemics plants of eastern Caucasus
1
Murtazaliev, R 1 , Asadulaev, Z 1
Mountain Botanical Garden of DSC RAS
The Caucasus is one of the 25 “hotspots” of biodiversity on the planet where more than 25% of plant species are endemic. In this regard one of the most original parts is the Central Daghestan. The long isolation, the peculiarities of orogenic process, the complexity of the topography and general aridity of the climate contributed to intensive micro-evolutionary process that involved elements of different floristic regions. 211 endemic species belonging to 37 families were identified in the
Eastern Caucasus. Most species belong to the Asteraceae family (38 species), 24 species belong to the Fabaceae family, 17 species to the Rosaceae family and 15 species belong to the Poaceae and Caryophyllaceae families. The richest endemic genera are Astragalus (13 species), Rosa
(1912), Campanula (11) and Psephellus (10 species).
This area is characterized by three monotypic endemic genuses: Pseudobetckea caucasica (Boiss.) Lincz.
(Valerianaceae), Mandenovia komarovii (Manden.)
Alava (Apiaceae), Muehlenbergella oweriniana (Rupr.)
Feer (Campanulaceae). The largest number of endemic species are concentrated in the middle mountain zone -
119 species (56% of the total number), less (about 40% -
84 species) is observed in the upper mountain zone. In the lower mountain zone there are only 43 species (20%).
The nature of the confinement of endemic species to different habitats shows that the greatest number of them is observed on stony and gravelly areas - 153 species (72,
5%). There are 4 centers of endemism in the Eastern
Caucasus . The largest is the Central Daghestan center with 112 species ( Allium mirzojevii Tscholok.,
Asplenium daghestanicum Christ, Àstragalus fissuralis
Alexeenko, À. daghestanicus Grossh., Ñentaurea awarica
Tzvel., Paederotella daghestanica (Trautv.) Kem.-Nath.,
Seseli alexeenkoi Lipsky, Scabiosa gumbetica Boiss.,
Tanacetum akinfievii (Alex.) Tzvel., etc.). Next by the number of species is the Eastern Caucasian center with
43 species: Trifolium raddeanum Trautv., Veronica bogosensis Tumadz., Erysimum babadagensis Prima,
Myosotis schistosa A.P. Khokhr., Vicia larissae Prima and others. The Albanian (26 species) and Iberian (11 species) centers includes a small number of species. A number of species on the northern macroslope is characterized by the confinement to the certain habitats but they cannot be assigned to the above mentioned centers. These species are much found in the foothills of
Daghestan and the middle belt of Chechnya and
Ingushetia. These are endemics with different origins and phylogenetic relationships: Allium grande Lipsky,
Campanula galushkoi (Taisumov et Teimurov)
Murtazaliev, Corydalis tarkiensis Prohk., Hornungia angustilimbata Dorofeev, Ferula calcarea Pimenov etc .
This work was supported by the Program of Presidium of
RAS 'Biodiversity' .
P0903 – ePoster
2
1
Phylogeny and classification of flax lilies (
Dianella
;
Xanthorrhoeaceae)
Muscat, KM 1 , Bayly, MJ 1 , Carr, GW 2
School of Botany, University of Melbourne, Australia;
National Herbarium of Victoria, Australia
623

Phylogeny and classification of flax lilies ( Dianella ;
Xanthorrhoeaceae) The flax lilies (genus Dianella ) have bedevilled taxonomists for over two centuries, largely due to overwhelming variation observed between species. Today, 40 taxa are recognised; potentially many species are yet to be named; some estimates are as high as 350 species . The greatest species diversity is in
Australia (22 species, 45 taxa), where the genus occurs from subalpine to coastal areas in rainforest, heathlands, rock outcrops and eucalypt woodlands . Elsewhere
Dianella occurs across Southeast Asia, the Pacific (to
Hawaii), New Zealand and Madagascar . This study presents a preliminary phylogenetic analysis of Dianella based on chloroplast sequence data . This phylogeny is being used to identify species groups and investigate the biogeography of the genus . The classification of species groups in Australasia is being further investigated using a combination of field studies, plant morphology and genetic markers. In particular, morphometric analyses of the Dianella tasmanica complex have suggested the existence of three undescribed species, and morphometric and sequence data are being used to assess the taxonomy of the Dianella caerulea complex (with nine varieties currently recognised) .
P0904 – ePoster
When spores lead us astray: molecular data supports abandoning the spore-number based subgenera in
Menegazzia
(Parmeliaceae, lichenized ascomycetes)
Myles, BC 1, 2 , Waters, JM 3 , Orlovich, DA 1
1 Dept of Botany, University of Otago, Dunedin, New
Zealand; 2 Allan Wilson Centre for Molecular Ecology &
3 Evolution, Palmerston North, New Zealand; Dept of
Zoology, University of Otago, Dunedin, New Zealand
Menegazzia is a genus of ca. 80 lichen species found mainly in Australasia, Melanesia and southern South
America . Most members of the genus have been placed into one of two subgenera based on the number of spores produced in the ascus . Eight spores per ascus places a species in subgenus Octospora R.Sant. (1942), and two spores per ascus places a species in subgenus Dispora
R.Sant. (1942) . Here, DNA data were used to assess the utility of this morphological character in delimiting
'natural' subgenera . Multiple collections from each of five Menegazzia species with 8-spored asci, five species with 2-spored asci, and three species which have not been known to reproduce sexually, were selected for
DNA sequencing . The nuclear ITS, 28S and EF1A loci, as well as the mitochondrial 12S, were sequenced across
54 individuals . The resulting partitioned dataset was then analysed phylogenetically within Bayesian, maximum likelihood, and maximum parsimony frameworks .
Coalescent-based Bayesian species tree, and parsimonybased phylogenetic network approaches were also employed for comparison . The results show all topologies from each method having the subgenus
Octospora as polyphyletic, and the subgenus Dispora as paraphyletic. The evolutionary history of the ascus in
Menegazzia therefore appears to be more complex than the original classification system suggests, with 1–3 independent transitions to dispory from octospory, or a single transition to dispory followed by a reversal to octospory in one clade . We conclude that continued use
624 of these two subgenera is not desirable, as they are not concordant with the inferred evolutionary history of the group . We recommend the abandonment of subgeneric rankings until a thorough revision of the genus has been completed.
P0905 – ePoster
Evolution of floral traits of the genus
Lonicera
L.
1
(Caprifoliaceae) distributed in Japan
Nakaji, M 1 , Sugawara, T 1
Makino Herbarium, Tokyo Metropolitan University,
Japan
The genus Lonicera (Caprifoliaceae) is mainly distributed in the Northern Hemisphere, and contains approximately 180 species . Among them, 23 species occur in Japan . Their flowers are usually actinomorphic or zygomorphic, and considerably vary among the species in corolla-tube length, orientation, and color .
Therefore, we suspect that the Lonicera flowers may have evolved in an intimate association with their pollinators. However, information available for the relationship between floral diversification of the
Lonicera species and their pollinators is restricted only a few reports . In addition, there still remains unclear as to the phylogenetic relationship among the species distributed in Japan . In the present study, We examined
Japanese Lonicera taxa (1) to clarify their molecular phylogenetic relationships, (2) to infer the evolutionary change of some floral traits among the taxa, and (3) to understand the pollination of some representative taxa .
The results obtained are as follows . (1) Molecular phylogenetic relationship of the Japanese Lonicera taxa based on chloroplast DNA. We performed Baysian phylogenetic analyses using combined cording regions and non-cording regions for 30 taxa of Lonicera occurring in Japan and one outgroup from the related genera, and obtained eight well-supported clades . (2)
Floral diversification and evolutionary change of floral traits in Japanese Lonicera taxa. We traced the evolutionary changes of flower symmetry, corolla-tube length, and orientation of flower in the phylogenetic tree based on chloroplast DNA. To trace the evolutionary change of floral traits, we used Mesquite ver. 2.72
(Maddison and Maddison 2009). In the phylogenetic tree, it is suggested that flowers are changed from actinomorphic to zygomorphic; corolla-tube with about
10mm long is the ancestral condition and longer or shorter corolla-tubes are derived independently; flowers hanging down is ancestral and flowers turning upward or sideways derived . It is also notable that the evolutionary changes of the above three traits occur at the same branch in the phylogenetiic tree . 3) Pollination in Japanese
Lonicera species. Flower visitors were mainly observed on 7 species occurring in Nagano and Yamanashi
Prefectures, central Honshu of Japan . Among the species examined, insects of hymenopteran species mainly visited the flowers in common and thus they appeared to be a useful pollinator . Based on the flower visitors, the species examined were divided into two groups: one is a group pollinated by only one or a few hymenopteran species, and the other group pollinated by various kinds of hymenopteran species . In the former group, species bearing actiomorphic or zygomorphic flowers are

included, and thus the floral symmetry appears to be unrelated with the kind of flower visitors.
P0907 – ePoster
The Commelinaceae of India: diversity and phytogeography
Nampy, Santhosh 1
1 Dept of Botany, St Joseph's College, Calicut-8, Kerala,
India
India is rich in Commelinaceae with 88 species in 13 genera . Eighteen species (ca. 20 %) are endemic to the region . Southern peninsular India and the foot hills of
Himalayas are one of the centers of diversity for
Commelinaceae . India can be classified into 11 phytogeographic zones: north west Himalayas, Indo
Gangetic plain, eastern Himalayas, Assam, central India, arid zone, northern Western Ghats and northern West
Coast, southern Western Ghats, southern West Coast and
Lakshadweep, Deccan, Eastern Ghats and Coromandel
Coast, Andaman and Nicobar islands . Of them, the
Western Ghats, Himalayas and Assam are rich in
Commelinaceae. The present paper discusses the diversity, endemism and phytogeographic pattern of
Indian Commelinaceae.
P0908 – ePoster
Biodiversity study of Cyanophyceae of Malangaon
1
Dam of Dhule of Maharashtra (India)
Nandan, S 1 , Ansari, Z 2
Dr PR.Ghogrey Science College, Dhule (M.S), India; 2
PH Mahila Mahavidyalaya, Malegaon, Dist. Nashik
(M.S), India
The Malangaon Dam is situated at Malangaon Village of
Sakri Taluka of Dhule district of Maharashtra. The dam is 20˚ 05’ N latitude & 74˚05’ E longitude. The catchment area of dam is 84–55 sq km. The storage capacity of dam is 13.03 m.cu.m Fresh water algal samples were collected at monthly intervals from 6 stations of Malangaon Dam of study area. The floating and attached epiphytic forms of algae were collected separately in acid washed bottles and preserved in 4% formalin for further taxonomic investigation. Line drawing of different different forms of Cyanophyceae were made by camera lucida. The systematic account of algal taxa of Cyanophyceae class was made for one year.
In present the study class Cyanophyceae mainly consists of 63 species belonging to 21 genera, viz. Microcystis,
Chroococcus, Gleocapsa, Gleotheceae, Aphanocapsa,
Aphanothece,
Dactylococcopsis,
Synechocystis,
Dermocarpa,
Merismopedia,
Myxosarcina,
Hydrococcus, Oscillatoria, Phormidium, Lungbya,
Symploca, Cylidrpspermum, Nostoc, Anabaena,
Calothrix, and Gleotrichia were found at study area. The dominant genus of Oscillatoria was showed with 19 species as compared to all other genera in present study .
P0910 – ePoster
Genetic structure and eco-physiology of the red mangrove hybrid zone (
Rhizophora mangle
and
R. racemosa
) in the Panama´s eastern Pacific coast
, Feliner, GN 2 , Turner, B 2 , Bermingham,
1
Ceron-Souza, I 1
E 2
Smithsonian Tropical Research Institute, Washington
DC, USA; 2 Real Jardín Botánico, CSIC, Madrid, Spain
Hybridization and introgression are common processes in both animals and plants. In plants, introgression of genes between species provides a pathway for the exchange of potentially adaptive variation, which promoted our interest in the potential role played by hybridization in the evolutionary history of Rhizophora mangle and R. racemosa , the two red mangrove species in Neotropics .
We found genetic evidence of ancient and ongoing introgressive hybridization between sympatric R. mangle and R. racemosa. However we still do not understand what is the genetic structure of the hybrid zones in which ongoing hybridization occur as well as the ecophysiological factors maintain this hybrid zone . It has been hypothesized that R. racemosa is less tolerant to high salinity compared with R. mangle and that this ecophysiological difference could explain how these species maintain their morphological and genetic distinctiveness in the face of gene flow . In order to test this hypothesis, we are analyzing a hybrid zone of red mangrove localized in the Panama’s eastern Pacific coast combining genetic (microsatellite markers) and ecophysiological information . The morphological and genetic identification of individuals showed a clear spatial zonation between species across three intertidal zones (i.e. shoreline, middle and inner) . In the shoreline, the dominant species is R. mangle (87.5%) with few individuals occupying middle zone (12.5%) and none in the inner zone . In comparison, the inner zone is dominated by R. racemosa (62%), with some individuals in the middle (38%) and none in the shoreline . This mutual exclusion was contrasted by the spatial distribution of hybrids . Hybrids are distributed across all intertidal zones, but in higher proportion in the middle
(75%) compared with shoreline (17%) and the inner zone
(8%) . Across seven months of study from May 2010 to
January 2011 we found strong phenological differences between species and hybrids (i.e. presence of flowers and fruits), suggesting that pre-mating phenologic isolation could help to maintain R. mangle and R. racemosa identity in the hybrid zone . In addition, the spatial distribution of species and hybrids appears to be related to a gradient of edaphic variables across intertidal zones.
Multivariate analysis of four edaphic variables (redox potential, salinity, pH and temperature) at depths of 10 and 40 cm showed an edaphic gradient across intertidal zones, especially in the redox potential (i.e. the amount of oxygen) and salinity . The salinity and the anoxia are higher in the inner zone, where R. racemosa is dominant, than in the middle zone, where hybrids are dominant, than in the shoreline, where R. mangle is dominant .
Although salinity tolerance across species seems to be contrary to our expectations, our results suggest that red mangrove hybrid zones are consistent with a hybrid superiority model . This model describes a strong environmental gradient where hybrids genotypes are less
625

fit in parental habitats and parental genotypes are less fit on areas relative to hybrids in the hybrid zone . We are conducting greenhouse experiments to corroborate this preliminary result.
P0911 – ePoster
Variation and functional differentiation of duplicated alcohol dehydrogenase gene in diploid plants of
Asteraceae
Nishino, T 1 , Nishiuchi, M 1 , Yahara, T 2
1 Osaka Prefecture University, Osaka, Japan; 2 Kyushu
University, Fukuoka, Japan
In plants, the alcohol dehydrogenase (ADH) is generally associated with the ethanol fermentation, and continued interest in evolutionary biology. ADH has been extensively studied in some plant groups as a model of multigene family with a relatively low copy number and these studies revealed that duplications of the Adh gene family occurred in multiple lineages. The Adh gene is a stress-inducible gene in plants. On the other hand, expression of ADH isozymes often depends on the organs. In addition to the diverse functions between duplicated Adh genes, recent studies have revealed the regulatory mechanisms causing the different expression of the genes. However, it remains uncertain whether multiple duplications of Adh gene family contributed to environmental adaptation through functional divergence of duplicated genes. To determine how duplicate gene copies can diverge in function and then act as a source of molecular adaptation, we examined ADh expression in diploid species of Asteraceae. The parameters of molecular evolution of cDNA sequences were estimated from organ-specific expression phenotypes of adh gene.
Isozyme numbers, organ specificity, and inducibility were evaluated for 29 taxa of Asteraceae under anaerobic condition. The electrophoretic results showed variation in number of isozyme, ranging from two to five. The mapping of isozyme number on the phylogenetic tree revealed that three loci as the ancestral state in the
Asteraceae family. The expression of ADH is highly variable in Asteraceae, and even within the genus Aster , suggesting the gene duplication and subsequent functional divergence of the multigene family. Variation of cDNA sequences was determined of mRNA of nonduplicated and duplicated ADH that was characterized as organ specificity and inducibility. Although the present evolutionary parameters data from cDNA sequences of
ADH are not sufficient, the debate about rejection of neutral selection of duplicated ADH genes remains.
P0912 – ePoster
Epidermal morphology of African Sapindaceae
Ogundipe, O 1 , Adeyemi, T 1 , Olowokudeje, JD 1
1 University of Lagos, Nigeria
Sapindaceae is one of the most important plant families in the tropics particularly in Africa most widely known for their use in medicine. Foliar epidermal studies were carried out on some representative members of the family
Sapindaceae in Africa. Fifty (50) taxa representing a total
626 of fifteen (15) genera were studied and illustrated with scanning electron microscopy (SEM). This revealed that the family exhibits a diverse range of characters, with no one feature being diagnostic. Most members of the family are hypostomatic. They exhibit anomocytic or paracytic stomata types; simple, non-glandular, glandular or stellate trichomes and straight, wavy or curved anticlinal wall patterns. Papillae are present in some members while others possess striae. A brief description of the epidermis of all genera studied is given, and character combinations are discussed which may allow the identification of these taxa.
P0913 – ePoster
A first attempt to the knowledge of relationships in the herbaceous bamboos (Poaceae, Olyreae): molecular phylogeny of
Raddia
and its allies based on noncoding plastid and nuclear spacers
4
5
1
Oliveira, RP
SH 3
University; 3
1 , Clark, LG
, Longhi-Wagner, HM
2 , Schnadelbach, A
4 , van den Berg, C
Universidade Federal da Bahia, Brazil;
5
2
1 , Monteiro,
Yowa State
Universidade Federal de Sergipe, Brazil;
Universidade Federal do Rio Grande do Sul, Brazil;
Universidade Estadual de Feira de Santana, Brazil
Raddia belongs to the tribe Olyreae and is almost completely restricted to forests in eastern Brazil. All nine accepted species are found in this region, with only one extending its distribution to extra-Brazilian areas, in northern and northwestern South America. The internal transcribed spacer (ITS) region of the rDNA and the noncoding spacer trn D-T of the cpDNA were sequenced for 37 samples from all species of Raddia and allied genera of herbaceous bamboos (tribe Olyreae, Poaceae), as well as members of woody bamboos (Bambuseae), in order to examine their relationships. The sequences were analyzed using maximum parsimony and Bayesian methods. Both the individual and combined analyses of
ITS and trn D-T supported Olyreae as monophyletic. All species of Raddia formed a well-supported monophyletic group, but without internal resolution. Individual analyses indicated incongruence regarding the sister group of Raddia , ITS analysis indicating Raddiella esenbeckii whereas trn D-T pointed out to Sucrea maculata . Combined analyses support Sucrea as a monophyletic genus sister to Raddia . Parodiolyra and
Raddiella were paraphyletic in all analyses, and Olyra as well, with some taxa interspersed among species of
Lithachne , Arberella and Cryptochloa . Eremitis and
Pariana appeared as an isolated clade within the tribe, and the position of Buergersiochloa remains uncertain.
P0914 – ePoster
Evolution of unisexual flowers and andromonoecy in
Pseuduvaria
(Annonaceae) inferred using molecular phylogenetics and pollination ecology
Pang, CC
RMK 1
1 , Su, YCF 1 , Scharaschkin, T 2 , Saunders,
1 University of Hong Kong, China;
University of Technology, Australia
2 Queensland

The Annonaceae is a large, early-divergent family of angiosperms consisting of c. 130 genera and 2,500 species. As with most early-divergent angiosperms, the majority of Annonaceae species have hermaphroditic, protogynous flowers. The temporal separation of the pistillate and staminate phases in Annonaceae flowers is reinforced by the existence of a non-receptive interim phase separating the two sexual phases. Although protogyny is a very effective mechanism to prevent autogamy, genetically-controlled self-incompatibility mechanisms appear to be lacking in the Annonaceae.
Several alternative mechanisms have been reported in the family to prevent geitonogamous self-pollination and hence inbreeding depression, including intra- and interindividual phenological synchrony and heterodichogamy.
The genus Pseuduvaria (56 species) is of particular interest as the majority of species (52) have unisexual flowers, although this is manifested in a number of different forms. Amongst the 41 species that are adequately known, 31 (76%) are andromonoecious, seven (17%) are completely monoecious; in contrast, only three species (7%) possess solely hermaphroditic flowers. The occurrence of staminate flowers has been inferred as the ancestral condition in the genus. In a recent monograph of the genus by Su & Saunders (Syst.
Bot. Monogr. 79:1-204. 2006), the structurally hermaphroditic flowers in the species that also possess staminate flowers were considered to be functionally pistillate. This conclusion was based on the observation that the stamens in the structurally hermaphroditic flowers produced small, irregular pollen that was likely to be sterile (observed, for example, in Pseuduvaria macrocarpa ); these stamens were therefore regarded as staminodes. A detailed field-based study of the reproductive biology and floral phenology of an endemic
Australian species, Pseuduvaria mulgraveana , was carried out in north-eastern Queensland, in October and
November 2010. Pollen viability tests demonstrated that the pollen grains in the stamens of the structurally hermaphroditic flowers was fertile, with equivalent levels of germination in vitro as pollen from staminate flowers.
The hermaphroditic flowers of P. mulgraveana were nevertheless shown to be functionally pistillate due to the dehiscence of the anthers c. 48 h after petal abscission.
This delayed anther dehiscence (or premature corolla abscission) therefore renders the pollen non-functional as the small pollinating beetles are not attracted to the flower and therefore do not collect pollen. This observation enables a reassessment of the evolution of floral unisexuality in Pseuduvaria , in which andromonoecy is determined as the ancestral condition in the genus, with several independent evolutionary origins of true monoecy and solely hermaphroditic flowers. An extended study based on Pseuduvaria species from different evolutionary lineages is currently underway.
P0915 – ePoster
Molecular phylogenetic studies on tropical species of
Lecanora
sensu stricto (Lecanorales: Ascomycota)
Papong, K 1 , Boonpragob, K 2 , Lumbsch, HT 3
1 Mahasarakham University, Thailand;
University, Thailand; 3
2 Ramkhamhaeng
Field Museum, USA
The genus Lecanora is one of the largest genera of crustose lichens and includes a number of distinct clades.
Lecanora sensu stricto is characterized by oxalate crystals in the apothecial margin and the presence of atranorin and/or usnic acid. Currently the phylogenetic relationships of tropical taxa in Lecanora s.str. are poorly understood. To address the phylogeny of tropical species and to test whether certain characters found only in tropical taxa (such as dark hypothecium ) have evolved several times or not. Molecular markers included two loci: the ITS region and the mitochondrial SSU rDNA.
Our analysis suggests that species with an amphithecium containing small crystals, viz. L. allophana , L. caesiorubella , L. campestris , L. carpinea , L. farinacea ,
L. imshaugii , L. intumescens and L. toroyensis form a monophyletic group. Species that contain usnic acid; viz.
L. achroa , L. elatinoides , L. queenslandica , L. pseudogangaleoides ssp.
verdonii , L. wilsonii and L. ulrikii do not form a monophyletic group, suggesting that the presence of this metabolite is of restricted taxonomic importance above the species level. Species with a dark pigmented hypothecium, viz. L. flavoviridis , L. phaeocardia , L. vainioi and Lecanora sp. C do not form a monophyletic group as well
P0919 – ePoster
The efficiency of RAPD and SSR markers in estimation genetic diversity of wild cranberry
(
Vaccinium oxycoccus
L.)
Paulauskas, A 1 , Zukauskiene, J 1 , Cesoniene, L 1
1
Daubaras, R 1
Vytautas Magnus Unicersity, Lithuania
,
In the last century during intensive peat bogs drainage, regulation of water levels, intensive cranberry picking has caused risk for wild cranberry Vaccinium oxycoccus population’s survival in Lithuania. Genetic variation represents the starting point for further evolution and is an important prerequisite for the prediction of evolutionary responses. This is of practical significance in the light of human-caused habitat fragmentation, alteration, or destruction. Genetic structure among and within populations also depends on the life history of a species. The present work was conducted to evaluate
RAPD (random amplified polymorphic DNA) and SSR
(Simple Sequence Repeat) marker systems for their ability to detect genetic diversity within and among V. oxycoccus populations and to compare the efficiency of these two marker types. Using nine RAPD primers we investigated genetic variation among and within 151 individuals (7 populations of different size Lithuanian and 3 populations of other countries) of V. oxycoccus .
With these profiles we evaluated 254 fragments.
According to RAPD results were selected different individuals 56 of V. oxycoccus and tested them with 10
SSR primer pairs. These SSR primers generated 62 fragments. According to RAPD data genetic variation among populations varied from 7 to 25 % (RAPD) and from 16 to 14% (SSR) molecular variance [AMOVA]).
The higher level of polymorphism detected by RAPD markers has contributed to the lower genetic similarity estimates based on RAPD markers as compared to SSR markers. The dendrograms generated with hierarchical
UPGMA (Unweighted Pair Group Method with
627

Arithmetic mean) cluster analysis revealed mixed and homogenous V. oxycoccus lineages. These lineages confirm the prediction that Lithuanian populations were derived from one population before glaciations.
P0920 – ePoster
Above and below-ground specialisation in Australian orchids: novel insights from integrated ecological and genetic analysis
Peakall, R 1 , Griffiths, K 1
R 2
, Linde, C 1 , Poldy, J 2 , Barrow,
1 Research School of Biology, Australian National
University, Canberra, Australia; 2 Research School of
Chemistry, Australian National University, Canberra,
Australia
Flowering plants are inextricably linked with other organisms. Above ground, orchids are well known for their diverse animal-pollination mechanisms. Below ground, orchids depend on mycorrhizal associations for seed germination. These interactions are often obligate and specialised and have likely played major roles in driving the astonishing diversification of the
Orchidaceae. One serious research impediment is our inability to identify species by morphology alone, in all three groups. Therefore, we are employing chemical, ecological and genetic tools to better understand these interactions. In sexually deceptive Chiloglottis orchids this work has revealed the novel chemistry used by cryptic orchid species to lure their male pollinators to the flower. Pollinator choice experiments have further confirmed the primacy of chemistry in controlling pollinator specificity. We have also uncovered multiple cryptic species of pollinator. Contrary to expectations, multiple species of orchid are associated with the one fungal taxon. Thus, pollinator, rather than fungal specialisation may have been the major driver of speciation in this group. These specific pollinator interactions may increase conservation risk for some orchid species. Our integration of chemistry, ecology and genetics is offering novel insights into orchid diversification and vulnerability that would not be possible from each field alone.
P0921 – ePoster
Sexy survival strategy:
Drakaea
orchids intercept wasp communication channels
Jeffares, L 1 , Bohman, B 1 , Phillips, R 2 , Peakall, R 3
1 Research School of Chemistry, Australian National
University, Canberra, Australia; 2 Kings Park and
3 Botanic Garden, West Perth, Australia; Research
School of Biology, Australian National University,
Canberra, Australia
Orchids are well known for their extraordinary species diversity and their specialised pollination systems. One intriguing pollination strategy is sexual deception, where the flower attracts male pollinators through mimicry of insect sex pheromones. All species of Drakaea , a genus endemic to southwestern Australia, are pollinated by the sexual deceit of male thynnine wasps. As part of a larger
628 study to better understand the ecology, evolution and conservation needs of Drakaea , an investigation into chemical communication between orchids and their pollinators is underway. Our work has focused on the three common species, D. glyptodon, D. livida and D. thynniphila . The volatile compounds produced by these flowers have been sampled by solvent extraction and solid phase microextraction (SPME) and then analysed using GC/electroantennographic detection (GC/EAD) and GC/MS to determine the bioactive compounds.
Further chemical analysis has revealed that several semiochemicals used by Drakaea flowers to attract their pollinators possess a pyrazine skeleton. This is an entirely different chemical system to that used in the closely related orchid genus Chiloglottis and other sexually deceptive orchid genera such as Ophrys. In D. glyptodon a blend of at least five physiologically active compounds appear to be involved in pollinator attraction.
Four of the compounds have been successfully identified, synthesised and proven to be attractive to pollinators in the field. We have strong indications that all species are utilising related compounds. Identification of those compounds is ongoing with the aim of determining the role chemical changes have played in speciation of
Drakaea . Further, this chemical knowledge may aid conservation efforts. Translocation and reintroduction of rare Drakaea will only succeed if selected sites are home to the orchid specific pollinators. Armed with synthetic pheromones to lure the pollinator it will soon be possible to more effectively survey pollinator availability and so improve site selection for Drakaea conservation.
P0922 – ePoster
Bulb and bulbotuberiferous plants of the Ukrainian natural flora: taxonomy, geography and conservation
1
Peregrym, M 1
O.V. Fomin Botanical Garden of the National Taras
Schevchenko University of Kyiv, Ukraine
Ukraine is one from biggest European countries.
Territory of country is situated in borders of three physiographical zones (mixed forests or Polissia, foreststeppe and steppe) and two montane regions (Crimea
Mountains and Carpathian Mountains). Diversity of nature conditions gives opportunities for growth about
5000 species of vascular plants in flora of the country.
There are from 129 (Cherepanov, 1995) till 138 species
(Mosyakin, Fedoronchuk, 1999) or about 2,6% of bulb and bulbotuberiferous plants in the Ukrainian natural flora. These plants are in structure of 6 families
(Alliaceae, Amaryllidaceae, Hyacinthaceae, Iridaceae,
Liliaceae, Melanthiaceae) and 22 genera. More presented genera are Allium L. (39 species), Gagea Salisb. (29 species), Ornithogalum L. (14 species), Crocus L. (9 species) and Tulipa L. (8 species). But, it is important to note that these genera excluding Crocus are very problematic in taxonomical aspect and it needs in future detailed analysis in the flora of Ukraine. Results of studying of geographic distribution of bulb and bulbotuberiferous plants in Ukraine show that increase of number of families, genera and species is observed from north to south and from foothills to peaks of mountains.
Main ecological factors determining such distribution are continentality, amount of precipitations and spring

average temperatures. Separately notice that 11 species bulb and bulbotuberiferous plants in the flora of
Ukrainian steppes and Crimea are endemic of narrow territories ( Colchicum fominii Bordz., Gagea aipetriensis
Levichev, G. hypanica Sobko, Ornithogalum melancholicum Klokov ex A. Krasnova and others).
Besides, part of north border of distribution of autumnflowering bulb and bulbotuberiferous plants is situated in
Ukraine. Majority of bulb and bulbotuberiferous plants of the Ukrainian natural flora are under state and international protection: 54 species are included in the
Red Data Book of Ukraine (2009), 10 species in the
IUCN Red List, 11 species in the European Red List and
4 species in Annexes of Bern Convention. Main causes of rarity for majority bulb and bulbotuberiferous plants are powerful anthropogenic influence on natural ecosystems, especially steppes, and also big popularity in use by local population of it for bouquets and transplantation from its natural habitats to domestic front gardens or graves. Studying of bulb and bulbotuberiferous plants as concrete life form will be continuing in the flora of Ukraine. But, international support of investigations for more efficiency is need, especially in taxonomical researches.
P0924 – ePoster
Biogeography and evolution of the African sweet vernal grasses (
Anthoxanthum
L. Poaceae; Pooideae)
Pimentel, M
Brochmann, C 3
1 , Sahuquillo, E 1 , Nemomissa, S 2 ,
1
2
Facultade de Ciencias, University of A Coruña, Spain;
National Herbarium, Biology Department, University of
Addis Abeba, Ethiopia;
University of Oslo, Norway
3 Natural History Museum,
The genus Anthoxanthum L. in the wide sense (including
Hierochloë R. Br.) includes ~49 mostly temperate species, with only six of them growing entirely in Africa.
Two of these African species ( A. nivale K. Schum. and A. aethiopicum Hedberg) are restricted to afro-alpine areas from Ethiopia to Tanzania, whereas four ( A. ecklonii
(Nees ex Trin.) Stapf, A. tongo (Nees) Stapf., A. dregeanum (Nees) Stapf and A. madagascariensis Stapf) grow in Southern Africa, from the Cape Region to
Madagascar. The two African groups are apparently not closely related. Whereas the afro-alpine taxa are morphologically most similar to a European species complex in subgenus Eu-Anthoxanthum , the South
African taxa are placed in the otherwise mainly Asian subgenus Ataxia , although some of them have not been well characterized morphologically. The evolutionary relationships among the African species and their biogeographic history are largely unknown. Here we address the following questions: (i) Do the African species constitute a monophyletic group? Are morphological and genetic variation consistent? ii) What is the origin of the African species of Anthoxanthum ? Is this consistent with findings for other African plants, particularly grasses, with similar distributions? (iii) What is the biogeographic history of the South African and
Malagasy species of Anthoxanthum ? Can a NE-SW migration route be inferred for the genus in Southern
Africa? We are currently sequencing different plastid
(trnH_psbA; trnT-F) and nuclear (ITS, ETS) DNA regions in samples of European, Asian and African
Anthoxanthum taxa, including all the Sub-Saharan species. The evolutionary relationships among the species will be assessed using parsimony, maximum likelihood and Bayesian inference. Biogeographic analyses will be based on vicariance/dispersal methods
(as implemented in DIVA; Ronquist 1997) and dispersal/extiction/cladogenesis methods (as implemented in DEC; Lagrange, Ree & Smith, 2007).
Our preliminary analyses suggest that the African taxa form at least two distantly related clades in the genus, and thus that at least two different colonization events must be inferred to explain the distribution and variation of sweet vernal grasses in Africa. The results are largely consistent with previous studies based on morphology. In the south African group, close affinity is suggested between the SE African A. ecklonii and the Malagasy A. madagascariensis , whereas the Cape species A. dregeanum and A. tongo appear to constitute a highly differentiated group.
P0926 – ePoster
Phylogeography and genetic differentiation along the distributional range of the orchid
Epidendrum fulgens
: a Neotropical coastal species not restricted to glacial refugia
Pinheiro, F 1
Lexer, C 3
, Barros, F
, Cozzolino, S 4
1 , Palma-Silva, C 1 , Fay, MF 2 ,
1 Instituto de Botanica, Sao Paulo, Brazil;
Laboratory, Royal Botanic Gardens Kew, UK; 3
2 Jodrell
Dept of
Biology, Unit of Ecology and Evolution, University of
Fribourg, Switzerland; 4 Universita degli Studi di Napoli
Federico II, Dipartimento di Biologia Strutturale e
Funzionale, Napoli, Italy
Phylogeographic studies in the Brazilian Atlantic
Rainforest have only been conducted with species associated to forest habitats, suggesting that a significant component of the evolutionary history of taxa associated to grassland and sand dune plant communities has hitherto been largely overlooked. This study examines the phylogeography of the sand dune orchid Epidendrum fulgens , in order to identify major genetic divergences or disjunctions across the range of the species and to investigate genetic signatures of past range contraction and expansion events. E. fulgens occurs in south and south-eastern seashore vegetation along Brazilian
Atlantic Rainforest biome. We used nine nuclear and four plastid microsatellite loci to genotype 424 individuals from 16 populations across the distribution of
E. fulgens . For both sets of markers, we estimated genetic diversity and population differentiation, testing for a north-south gradient of genetic diversity. A haplotype network was inferred based on the plastid haplotypes.
For the nuclear markers, we used a Bayesian assignment analysis to infer population structure. Past demographic changes were tested using a coalescent approach. The microsatellite data sets suggest a low level of population differentiation (GST = 0.097) and low inbreeding coefficient (f = 0.079). There was no significant evidence of phylogeographic structure in either nuclear and plastid markers. Past demographic changes were detected in northern populations. Such demographic changes were not expected for those populations since previous studies
629

with forest species found evidence of refuges in the same areas. Higher genetic diversity was found in southern populations, a different pattern than observed in previous studies with forest-dwelling species. The results are consistent with the long-term persistence of E. fulgens in many regions where forest retraction and fragmentation were inferred for species associated with forest habitats.
Bottlenecks were detected in populations from areas where population expansion events were also detected previously for other animals and plant species, suggesting that forest expansion after the LGM played a role in population fragmentation and decrease in genetic diversity in this species. The expansion of grasslands habitats, as revealed by palynological data during the
LGM, apparently had a positive effect on this sand dune species.
P0928 – ePoster
Do ecotypes of
Rhinanthus alectorolophus
(Orobanchaceae) represent genetically distinct entities? A case study in southwestern Germany
, Esfeld, K 2 , Blattner, FR 1 , Thiv, M 3
1
Pleines, T 1
Leibniz Institute of Plant Genetics and Crop Plant
Research (IPK) Gatersleben, Germany; 2 Institute of
3
Plant Sciences, University of Bern, Switzerland;
Staatliches Museum für Naturkunde Stuttgart, Germany
European annual species of the genus Rhinanthus often exhibit seasonal ecotypic variation, with populations showing differences in flowering time and a number of correlated morphological characters. The ecotypic forms are mostly recognised as subspecies, but their value is disputed. The present study evaluates the correlation of morphological characters and genetic differentiation of populations of Rhinanthus alectorolophus . Thirty-nine populations of three different subspecies from the southwestern German federal state of Baden-Württemberg were sampled. A total of 798 individuals were used for morphological analyses, 187 of them also for molecular
AFLP analyses. In a discriminant analysis based on morphological characters, 89.7% of all individuals were correctly assigned to their previously determined subspecies, indicating that subspecies identification is ambiguous for some populations. Using AFLP data and the software Structure, the individuals could be assigned to three genetic clusters that do, however, not correspond to the subspecies. In most populations, all AFLPscreened individuals belong to the same genetic cluster, but some populations appeared admixed. The distribution of the three genetic clusters shows a clear geographic pattern. One cluster occurs in the upper Rhine Valley, the second one mainly in the western part of the
Schwäbische Alb, and the third cluster is found in the remaining study area. A Mantel test showed a significant correlation between genetic and geographic distances for the whole dataset. Correlations of genetic distances with single morphological characters are weak and mostly insignificant. A weak but possibly significant correlation was found between genetic distance and difference in the number of internodes below the inflorescence, the most important character for subspecies delimitation. Since this character is linked to flowering time, it might restrict gene flow between populations belonging to different ecotypes or subspecies. However, this impact does not
630 seem to be very important, since the main genetic structure found in the dataset is a geographical one. In summary, the results indicate that the ecotypic variation in R.
alectorolophus represents local adaptation to different habitats and that the subspecies do not form discrete entities.
P0929 – ePoster
Reinstatement and expansion of the southwest Pacific genus
Plerandra
(Araliaceae)
Lowry II, PP 1 , Plunkett, GM 2 , Frodin, DG 3
1 Missouri Botanical Garden, St Louis, USA;
Botanical Garden, USA; 3
2 New York
Royal Botanic Gardens, Kew,
UK
Recent molecular phylogenetic studies have shown that the pantropical genus Schefflera , long regarded as the largest in Araliaceae (with perhaps 800 or more species), is polyphyletic, comprising five clades scattered throughout the family . One of these, referred to as the
Melanesian Schefflera clade, includes ca. 100 species from several Archipelagos in the southwest Pacific (New
Caledonia, Fiji, Vanuatu, Solomon Islands and New
Guinea) . Some of these species have historically been assigned to Schefflera while others, placed in segregate genera such as Dizygotheca and Plerandra , were transferred to Schefflera during a process of dramatic expansion in the genus . An integrated program of field exploration, examination of herbarium material, and molecular phylogenetic analyses has led to the discovery of more than a dozen new species and has provided insights into relationships within the Melanesian
Schefflera clade . Five morphologically and geographically coherent subclades can be recognized, two of which correspond to Dizygotheca (distinctive among Araliaceae in having anthers with 4 thecae) and
Plerandra (whose members exhibit varying degrees of polymery, with up to 19 carpels and 500 stamens in some taxa), whereas the remaining subclades largely correspond to informal groups previously proposed on the basis of morphology and wood anatomy . Using nearcomprehensive taxon sampling, the results of our molecular analyses reveal multiple inter-island dispersal events, primarily from New Caledonia to other
Archipelagos, but also among other island groups .
Because the generic name Schefflera belongs to another clade, we have opted to place all of the species comprising the Melanesian Schefflera clade within an expanded Plerandra in order to provide a revised taxonomy that reflects the biogeographic, morphological and phylogenetic information now available . To retain the information of the phylogenetic structuring of the five distinctive subclades, we recognize these as subgenera – an approach recently adopted for Polyscias , the second largest genus of Araliaceae.
P0930 – ePoster
Pollination of
Plectranthus
(Lamiaceae) by acrocerid flies (
Diptera
: Acroceridae) in South Africa
Potgieter, C 1 , Edwards, T 2 , Van Staden, J 3

1 Bews Herbarium, School of Biological & Conservation
2
Sciences, University of KwaZulu-Natal, South Africa;
School of Biological & Conservation Sciences,
University of KwaZulu-Natal, South Africa; 3 Research
Centre for Plant Growth and Development, School of
Biological & Conservation Sciences, University of
KwaZulu-Natal, South Africa
At least five species of acrocerid fly are involved in the pollination biology of five species of Plectranthus , studied in the KwaZulu-Natal and Eastern Cape provinces of South Africa. These 'Small-headed flies' are nectar-feeding floral visitors to straight-tubed species that have corolla tubes of medium length (5–16 mm). They are important pollinators of two narrow Pondoland endemics, P.
oertendahlii and P. praetermissus , yet remain relatively unknown as pollinators in South Africa.
This paper illustrates the close relationship between the
Acroceridae and a number of Plectranthus species.
P0931 – ePoster
Taxonomy and phylogeny of the fern genus
Megalastrum
(Dryopteridaceae)
Prado, J
A 2
1 , Moran, R
, Hanks, J 4
2 , Labiak, P 3
, Schuettpelz, E 5
, Sundue, M
, Rouhan, G 6
2 , Vasco,
1 Instituto de Botânica, São Paulo, Brazil;
Botanical Garden, USA;
Paraná, Brazil;
York, USA;
USA; 6
4
3
2 New York
Universidade Federal do
Marymount Manhattan College, New
5 University of North Carolina Wilmington,
Muséum National d'Histoire Naturelle, Paris,
France
We conducted a worldwide taxonomic revision and phylogenetic analysis of Megalastrum , a genus of terrestrial ferns that grows in wet shady montane forests.
About 50 species were previously accepted in the genus; we now recognize about 100. Formerly, the genus was known only in the Neotropics and Africa-Madagascar, but we found it also occurs on several islands in the southern oceans. Phylogenetically, the genus consists of five main clades that correspond well to geography. Two species from the Mascarenes, which are characterized by the synapomorphy of fenestrate spores, are sister to the remainder of the genus, which consists of a clade of 1) primarily circumaustral species, 2) the ‘pulverulentum clade’ (consisting of species with leaves 2–4 m long), and 3) a large unnamed clade. The latter clade comprises two subclades, one including species endemic to southeastern Brazil, the other including species from
Central America, the Antilles, and the Andes. The latter subclade is characterized by the synapomorphy of spores with parallel crests (spiny spores are present elsewhere in the genus). Nested in the Brazilian clade is M. lanuginosum , the only African species; it represents a dispersal event from Brazil to Africa. Characters such as leaf division and the presence of indusia have been traditionally used in keys to divide the species in large groups; however, these characters have evolved independently within the genus. A website associated with the project may be found at http://sweetgum.nybg.org/ Megalastrum /index.php
(Funded by United States National Science Foundation
DEB 0717056).
P0932 – ePoster
Molecular phylogeny reveals a new genus of grammitid ferns (Polypodiaceae) from the Neotropics
Hirai, R 1 , Rouhan, G 2 , Labiak, P 3 , Ranker, T 4 , Prado, J 1
1 Instituto de Botânica, Sao Paulo, Brazil; 2 Muséum
3
National d'Histoire Naturelle Paris, France;
Universidade Federal do Paraná, Brazil; 4 University of
Hawaii, USA
We conducted a phylogenetic analysis of
Micropolypodium s.l., a genus of grammitid ferns that grow mainly as epiphytes in wet mountain forests.
Initially, the genus was known only from Asia (Japan), but this generic name was progressively applied to
Neotropical species, and about 30 species were assigned to this genus. However, our phylogenetic analysis, based on three molecular markers ( rbc L, atp B, and trn LF), showed that only three Asian and Malesian species belong to a Micropolypodium s.s. clade. The other 28 species formed a clade that is entirely Neotropical and which is sister to a clade of several Asian genera, including Micropolypodium s.s. A morphological synapomorphy for this Neotropical clade is the presence of minute and slightly catenate branched and unbranched hairs occurring together. Some hairs have a setiform branch and are mostly found on the petiole, costa, and laminar tissue abaxially. On the other hand,
Micropolypodium s.s. has just minute catenate unbranched hairs borne on the same parts of the leaf.
Based on these results, we recognize the Neotropical clade as a new genus of grammitid ferns, segregated from
Micropolypodium . There is no available generic name for this group and a new name will be proposed (Funded by
FAPESP, grant n. 06/06215-5).
P0933 – ePoster
Diversification and radiation of
Banksia
in the southwest Australian hotspot and beyond
Cardillo, M 1 , Pratt, R 1
1 Australian National University, Canberra, Australia
Mediterranean-climate biodiversity hotspots represent one of the most intriguing problems in ecology and evolution. Southwest Australia, for example, supports
8000 plant species, half of which are endemic, despite low primary productivity and a lack of significant topographic variation. We are using molecular phylogenies to understand why many plant genera have undergone massive radiations in the southwest compared to elsewhere in Australia. Here we present the first nearcomplete phylogeny of the genus Banksia, and use it to explore patterns of diversification in southwest and nonsouthwest clades.
P0934 – ePoster
Cretaceous angiosperms – an Indian perspective
Rajanikanth, A 1
631

1 Birbal Sahni Institute of Palaeobotany, Uttar Pradesh,
India
Global records of flowering plants suggest Cretaceous origins. Recent spurt in fossil records have thrown new light on early radiation of angiosperms. Early Cretaceous times witnessed invasion of basal angiosperms and their radiance subsequently directed their spread to different geographical locales. Fossil evidences from China, USA,
Russia, Israel, Portugal and other places redefined accepted inquiry and firmly established wide occurrence of angiosperms during early Cretaceous times. The
Gondwanan continents too witnessed early-late
Cretaceous explosion of this group. In this perspective recent finds of angiosperm fossils from the Indian east coast Gondwanan basin opened new possibilities to understand their spread in the southern continents.
Hitherto Indian evidences have been scarce and could not furnish enough support for an early Cretaceous radiation of flowering plants on the Indian continent. New evidences from the east coast Gondwana basin, viewed in conjunction with records from other Gondwana continents firmly establish angiosperm spread during the early Cretaceous times. Though they attained the level of ecological dominance during the Maastrichtian times, as evidenced by the Indian Deccan Inter-trappen flora, their spread in the east coast Gondwanan basins negate the accepted view of mutual exclusion of Ptilophyllum and angiosperm floras. Evidences found in the form of leaves, pollen, fruit have been evaluated to decipher coexistence of Ptilphyllum flora and angiosperm flora.
Prominence of fossil angiosperms is yet to be established in the diverse and prominent ecosystems of Indian Early
Cretaceous ecosystems. In the present communication
Cretaceous angiosperm records from India have been critically evaluated. The synthesis is integrated with latest global evidences to derive a meaningful inference.
Increasing diversity and complexity of the angiosperms through the later part of Cretaceous on the Indian continent has been attributed to unique palaeogeographic position occupied by Indian sub-continent during the
Cretaceous.
P0935 – ePoster
An investigation into the population diversity of
Borya mirabilis
and phylogenetics of the Boryaceae
Reiter, N 1 , Lawrie, A 2 , Walsh, N 3
1 Dept of Sustainability and Environment;
University, Melbourne, Australia; 3
2 RMIT
Royal Botanic
Gardens, Melbourne, Australia
Borya mirabilis is one of the world’s most critically endangered plants. This research has illuminated key aspects of its reproductive biology; interspecies and intraspecies molecular relationships. Each of these aspects has fundamental management implications for the active management of B. mirabilis. Floral observations of B. mirabilis and related species affirmed the uniqueness of the Borya ceae amongst the
Asparagales. B. mirabilis had an unusually high number of floral abnormalities compared with other species of
Borya observed. B. mirabilis is fly-pollinated. Pollen of
Borya species showed little difference in the characteristics of mature pollen between species, with
632 viable pollen being prolate and unicolpate with a single colpa-style aperture and a unique patterning of the pila.
The structural immaturity of B. mirabilis pollen correlated with evidence from pollen growth experiments, where B. mirabilis pollen had extremely low germination rates, with those grains that did germinate being slow to do so and with slow-growing pollen tubes compared to those of fertile Borya species.
Examination of the ovules of B. mirabilis showed that morphologically they were viable compared to viable
Borya species. The field population of B. mirabilis was crossed, with one seed produced (the first recorded seed for this species). Cross-pollination using the pollen of the closely related B. constricta and B. sphaerocephala with
B. mirabilis ovules proved unsuccessful. Examination of the chromosome number of B. mirabilis showed that it had approximately 66 chromosomes and is probably hexaploid, relative to the diploid number of 26 in B. constricta . This may explain its low fertility . Interspecies and intraspecies relationships of the Boryaceae and
Borya mirabilis were investigated using sequences of chloroplast and nuclear DNA. The closest similarities to
B. mirabilis were B. constricta and B. sphaerocephala. B. mirabilis may have emerged from alloploidy of these species in the past. Because of the consistent similarities of B. mirabilis and B. constricta chloroplast sequences, it is proposed that both shared a common ancestor with a chromosome number of 2n=22. A malfunction n meiosis may have resulted in ovules with 2n=44. The high similarity of the nuclear ribosomal ITS region DNA suggests that the nuclear DNA was derived from B. sphaerocephela . B. mirabilis may be an allopolyploid, from fertilisation of a diploid ovule of B. constricta with haploid pollen of B. sphaerocephala , resulting in a reproductively isolated polyploidy of low fertility. The wild population of B. mirabilis was determined to have a small amount of genetic variation. The genetic variation in the field population was not fully reflected in the ex situ population . This research has provided critical knowledge to aid the recovery team in its current and future endeavours to manage this species and bring it back from the brink of extinction.
P0936 – ePoster
Patterns of morphological variation within and between
Lejeunea
species have implications for alphataxonomy
Renner, MAM 1 , Brown, EA 1 , Wardle, GM 2
1 National Herbarium of New South Wales, Sydney,
Australia; 2 School of Biological Sciences, University of
Sydney, Sydney, Australia
Since the re-classification of the thallophytes, it has been recognized that the most obvious features of organisms are not necessarily the most meaningful in classification.
In the leafy-liverwort family Lejeuneaceae, subjective assessment of gametophyte size and shape is an important source of taxonomic characters, and in alphataxonomic studies these characters are coupled with a species concept based on morphological discontinuity.
We quantified gametophyte size and shape in four
Australasian species of Lejeunea using allometric and geometric morphometric methods. In these species the correlation between phylogenetic isolation and

morphological discontinuity did not hold. All four species exhibited morphological overlap in size and shape. Within all four species, populations exhibited significant differences in size and shape, and some pairs of populations occupied mutually exclusive regions of multidimensional size and/or shape space. As a result, some populations were more similar to those of other species, than to those of the same species. The phylogenetic isolation of the study species examined here within the genus Lejeunea suggests that the results are generalisable across the genus and we should not expect
Lejeunea species to be bounded by zones of phenetic morphological discontinuity. This implies that traditional taxonomic approaches to the interpretation of morphology may severely underestimate diversity. The same conclusion was reached by a recent molecular study of another genus belonging to the Lejeuneaceae.
P0937 – ePoster
Tropical forest bryophytes from Tabasco, Mexico
Rivas-Acuña, G 1 , Escolastico-Ortíz, D 1
1 Universidad Juarez Autonoma de Tabasco, Mexico
The state of Tabasco has a humid tropical climate adequate for bryophytes. Up to today there had not been enough studies especially lately for this group of plants, although, they are especially abundant and offer many environmental services like soil formation and humid retention. Due to the fact that the last two decades the state has suffered a strong degradation related to deforestation and natural hazards it is important to determine the species that are present in the area. The objective of the research was to make a diagnostic of the distribution, diversity and richness of this group of plants. Plant collections were performed on all the areas of study to gather information and map the actual distribution. Results show an important distribution on relicts of tropical and subtropical forest of the area as well as wetlands and secondary vegetation. There are 96 species from 50 genera and 20 families.
Calymperes ,
Octoblepharum , Fissidens , Funaria , Marchantia ,
Taxithelium , Trichosreleum – these genera are wide distributed, while Haplocladium , Syrrhopodon , are especially found on wetlands that are now important endangered areas. A special mention had to be done to the genus Erpodium that is present on Tabebuia rosea
(Bignoniaceae) that is an important representative species of the tropical forest of Mesoamerica.
P0938 – ePoster
New floristic discoveries and biodiversity of the western Canadian Arctic vascular plant flora
Saarela, JM 1 , Gillespie, LJ 1 , Consaul, LL 1 , Bull, RD 1
1 Canadian Museum of Nature, Ottawa, Canada
Exploration of the vascular plant flora of the Canadian
Arctic has been ongoing for almost two centuries, yet substantial gaps remain in our floristic understanding of this large and difficult-to-access region . Detailed information on the diversity and distribution of Arctic plants is urgently needed to understand the potential impacts of climate change on the region’s flora . We recently (2008, 2009, and 2010) conducted detailed floristic surveys in botanically-understudied regions of the western Canadian Arctic: on mainland Northwest
Territories (Tuktut Nogait National Park and vicinity, between the tree line and the Arctic coastline), and on
Victoria Island, the largest island in the western
Canadian Arctic Archipelago . The comprehensive baseline data of our >3000 collections adds important new knowledge to our understanding of Arctic plant biodiversity, and could facilitate understanding the impacts of global climate change on the composition and distribution of the Arctic flora. Many of our collections are first records for the northwestern Arctic, and we documented many range extensions for species within
Canada and the NWT (e.g., Botrychium lunaria ,
Equisetum palustre , Carex concinna , C. garberi , Draba oligosperma , Lomatogonium rotatum , and Myriophyllum sibiricum ) . Surprisingly, we found previously undocumented extralimital stands of balsam poplar
( Populus balsamifera ) growing fully in the tundra well beyond the reported range of the species in Canada, which is generally considered to extend only to the tree line . Dense ‘Arctic forests’ dominated by the large willow, Salix alaxensis , occur along river terraces on northwestern Victoria Island, forming a unique microhabit that harbours a lush and diverse vegetation atypical of the surrounding Arctic tundra. Through our explorations of Victoria Island, we have documented species not previously reported for the Canadian Arctic
Archipelago as a whole (e.g., Andromeda polifolia ), the western Canadian Arctic Archipelago (e.g., Koenigia islandica , Corallorhiza trifida , and Pinguicula vulgaris , representing a major northern range extension for this insectivorous species), and Victoria Island (some 30 species, including major range extensions for the recently-described grass species, Puccinellia banksiensis ).
P0939 – ePoster
1
DNA barcoding the vascular plant flora of the
Canadian Arctic
Saarela, JM 1 , Gillespie, LJ 1 , Consaul, LL
Canadian Museum of Nature, Canada
1 , Bull, RD 1
Arctic ecosystems are the most rapidly changing on the planet in response to global climate change. Accurate identifications of Arctic plant species are critical for monitoring, studying, and understanding potential climate-induced changes in their diversity, distributions, and community assemblages . To facilitate rapid identification of Arctic plant taxa, we are producing plastid DNA barcode data ( mat K and rbc L) for the entire
Canadian Arctic vascular plant flora . Preliminary analyses of data from more than 500 individuals representing some 225 species indicate that Arctic plant families and most Arctic genera are consistently and reliably differentiated with plastid barcode data . In line with the results of other plant barcoding studies, specieslevel discrimination varies dramatically among lineages .
For example, barcode data distinguish most of the Arctic species of Carex (including Kobresia ), taxonomically difficult plants which dominate vast tracts of Arctic tundra; this remarkable barcoding success is a function of
633

the (mostly) distant relationships among these Arctic sedge species . In the Brassicaceae, barcode data effectively distinguish the Arctic genera (e.g., Braya ,
Descurainia , Draba , Erysimum , Parrya ), but infrageneric variation is low and unable to distinguish most species, especially in the diverse and difficult genus
Draba where it would be most useful. Genera of
Ericaceae are clearly distinguished, and there is informative interspecific plastid variation in those genera with more than one Arctic species, including
Rhododendron and Vaccinium. Overall, plastid DNA barcoding performs quite well for Arctic plants when considered at a regional, eco-geographical level . A complete species-level barcode database will no doubt facilitate future ecological and systematic research on the
Arctic flora. Nevertheless, nuclear-based barcode markers will need to be developed to identify speciesspecific nucleotide differences in several difficult genera with minimal plastid variation, such as Salix
(Salicaceae), Taraxacum (Asteraceae), and Potentilla
(Rosaceae).
P0940 – ePoster
Phylogenetic position of the
Takakiopsida
(Bryophyta) inffered from comparative chloroplast genomics
, Shimamura, M 1 , Yamaguchi, T 1 , Sadamitsu, A 1
Deguchi, H 1
1 Dept of Biol. Sci., Grad. School of Sci., Hiroshima
University, Japan
An enigmatic moss genus, Takakia was originally described as a liverwort due to its primitive gametopytic feature. When the sporophyte of Takakia was discovered, it became obvious that it was a moss because it had a calyptra, columella and well-developed seta. However the dehiscence of capsule by a diagonal slit is unparalleled in extant mosses.
Takakia is now treated as an independent family, order or even class among the mosses as one of the most primitive taxon. There are several opinions on its phylogenetic position and additional data are required for better understanding of the genus. For resolving the plant phylogeny at deep levels, the chloroplast DNA sequences might be useful because of their lower rates of silent nucleotide substitution. Furthermore, structural characters of chloroplast genome, such as loss or gain of gene, inversions of gene order, and expansion and contraction of gene, have also been drawing attention as powerful phylogenetic markers. We analyzed the chloroplast genome sequence of T. lepidozioides , to elucidate the phylogenetic relationship among the major taxon of mosses including Takakiopsida. The chloroplast genome of T. lepidozioides is a circular DNA of about 150 kb and consists of a large single copy (LSC), a small single copy and two inverted repeat (IR) regions. The chloroplast genome of T. lepidozioides is slightly larger than those of other mosses and the gene content is similar to those of liverworts and hornworts. The genome contains at least
86 proteins, 32 tRNA, 4 rRNA, 4 open reading frames
(ORFs) and some pseudogenes. Among them, four protein genes (rpoA, cysA, cysT, ccsA) have been known in the chloroplast genomes of liverworts and hornworts, but absent in the Bryopsid mosses, Physcomitrella patens and Syntrichia ruralis . In addition, the ribosomal protein
634 small subunit 16 (rps16) which is unknown in Bryopsid mosses, is found in T. lepidozioides . In other primitive mosses, Andreaea nivalis (Andreaeopsida), cysA, cysT and ccsA are found as pseudogenes in the chloroplast genome. In Sphagnum palustre (Sphagnopsida), cysA, cysT and rps16 are found as pseudogenes in the chloroplast genome. Phylogenetic distribution of these genes in the chloroplast genomes might be effective markers for considering the relationships of major moss lineages. Structural characters of chloroplast genome of
T. lepidozioides support the monophyly of bryophytes and the basal position of the Takakiopsida among the mosses.
P0941 – ePoster
Phylogenetic relationships in the subfamily Pooideae
(Poaceae) based on combined three plastid DNA regions ( ndh
F, trn
Hpsb
A,
mat
K)
Sahuquillo, E 1 , Pimentel, M 1 , Catalán, P 2
1 Facultade de Ciencias, Universidade da Coruna, A
Coruna, Spain; 2 Escuela Politécnica Superior de
Huesca, Universidad de Zaragoza, Huesca, Spain
The subfamily Pooideae of temperate grasses is the largest worldwide distributed group of Poaceae (150 genera, 3300 species), with representatives spread in most of the biomes of the temperate zones in both hemispheres. The initial systematic proposals based on molecular analyses recognized 12 tribes in the Pooideae.
However this classification has varied in recent years due to the use of combined multi-gene analyses and the expansion of the taxonomic sampling. Despite this progress, the phylogeny of the subfamily is not completely resolved. Moreover, different inconsistencies have been found between the nuclear-based and the organellar-based topologies recovered . The aims of this study are: (I), to estimate the phylogenetic relationships among the major lineages of temperate grasses belonging to the Pooideae (Poaceae), (ii) to clarify their taxonomical structure/status and (iii) to evaluate the usefulness of combining cpDNA sequences data sets in obtaining more robust phylogenetic hypotheses. To address these aims, we use three plastid DNA regions
( ndh F, trn Hpsb A, mat K) that present relatively high rates of substitutions and transversions in the Poaceae.
Samples were selected to cover those taxonomic
Pooideae tribes and subtribes underrepresented in previous studies carried out with cpDNA. We will sequence 155 species belonging to 66 genera, representing the 12 tribes previously recognised in the subfamily. Furthermore, two representatives from subfamily Danthonoideae and Chloridoideae will be used as outgroups. The sequences obtained from these three regions will be analysed jointly in a combined data matrix. Gaps will be treated as binary characters following the simple indel coding method of Simmonds
& Ochotorena (2000). Molecular phylogenies will be attained using standard phylogenetic methods. First, a heuristic search for most parsimonious trees will be conducted with PAUP version 4.0b8 (Swofford, 2002).
Strict consensus trees will be computed from all equally most parsimonious trees and internal branch support will be estimated using non parametric bootstrapping. A model based on Bayesian inference will also be used to

reconstruct phylogeny in order to reduce the impact of model misspecification on the topologies recovered.
Preliminary results show topologies partially consistent with previous analyses based on plastid data. Ongoing analyses will allow us to assess the taxonomy and systematics of the different subgroups recovered.
P0947 – ePoster
Origin and evolution of New Caledonian
Diospyros
(Ebenaceae): a phylogenetic approach
Samuel, R 1 , Turner, B 1
Wallnöfer, B 4 , Chase, M 5
, Duangjai, S 2
, Barfuss, MHJ
, Munzinger, J
1
1 Dept of Systematics and Evolutionary Botany; 2
3 ,
Dept of
Forest Biology, Faculty of Forestry, Kasetsart
University, Bangkok, Thailand; 3 Laboratoire de
4
Botanique, Centre IRD de Nouméax, New Caledonia;
Natural History Museum, Vienna, Austria; 5 Jodrell
Laboratory, Royal Botanic Gardens, Kew, UK
Diospyros is one of the largest genera comprising approximately 500 species of which 31 are distributed in
New Caledonia. The recent molecular studies on family
Ebenaceae where a subset of New Caledonian Diospyros was included highlighted the presence of four lineages and two types of endemism. The first being paleoendemics, which suggested ancient Gondwanan
(Australian) origin, e.g., D. macrocarpa , D. brassica and
D. balansae ; and the second being neoendemics, elements coming relatively recently from the Indo-
Malaysian region that include species like D. vieilardi ,
D.
umbrosa and D. parviflora . This neoendemic species of New Caledonia group together with Hawaiian D. sandwicensis and widely distributed D. ferrea . The level of DNA sequence divergence among the neoendemic species is relatively low and does not appear to be correlated with the level of phenotypic diversity. The steep environmental gradients and unusual soil types in
New Caledonia appear to have facilitated speciation in this group of neoendemics. Species boundaries among them are unclear and under discussion by different authors. In terms of conservation priorities for New
Caledonian species, our results support the existence of four genetically distinct groups on this island. Each lineage of New Caledonian Diospyros should be treated as a separate conservation unit.
P0948 – ePoster
Phylogenetic relationships in subtribe Oncidiinae
(Epidendroideae, Orchidaceae): a combined approach.
Sandoval, E 1 , García, CJ 2 , Villaseñor, JL 1
1 Instituto de Biología, Universidad Nacional Autónoma
2 de México; Departamento de Botánica, Universidad
Autónoma Metropolitana, Mexico
Subtribe Oncidiinae (Family Orchidaceae, subfamily
Epidendroideae) is an important element in the
Neotropics, including Mexico. It includes about 95 genera and 1700 species and is considered one of the most diverse subtribes, both taxonomically and morphologically. Based on molecular systematics, the relationships among its members have been recently assessed, but structural characters have been poorly used in this issue, although their systematic relevance has been demonstrated in other groups of Orchidaceae. With the aim to evaluate the importance of structural characters in the phylogeny of Oncidiinae, a parsimony analysis including 57 species, representing 37 genera of this subtribe and other four additional subtribes (outgroups) was carried out. The analysis combined 2228 DNA characters (from ITS and mat K regions), in addition to 37 structural characters (29 anatomical and 8 morphological). DNA sequences were obtained mostly from the GenBank (except four species), structural data were obtained revising permanent preparations (dermal, vascular and fundamental tissues), and morphological data were obtained from the literature and observation of living specimens housed in several Mexican and foreign greenhouses. The parsimony analysis was carried out using Nona (under the platform of Winclada), considering all characters as unordered with equal weight; a heuristic search was performed and with the
Jackknife resampling technique the branchs support was obtained. We obtained two equally parsimonious trees that differ just in the placement of a single taxon. The results support the monophyly of the subtribe Oncidiinae, characterized by the presence of papillae on the abaxial and adaxial leaf surfaces. In addition, seven structural synapomorphies were recovered; they define five clades all them with high Jackknife support (>75%). Other clades are supported by structural characters as monophyletic ones, although such characters are resolved as reversals. This study points out the importance of structural characters in the phylogeny of Orchidaceae's groups, which resolve as very informative data along with the more currently used molecular data.
P0949 – ePoster
The phylogenetic relevance of mineral inclusions in the subtribe Oncidiinae (Epidendroideae,
1
Orchidaceae
Sandoval, E 1 , García, CJ 2 , Villaseñor, JL 1
Instituto de Biología, Universidad Nacional Autónoma de México; 2 Departamento de Botánica, Universidad
Autónoma Metropolitana, Mexico
About 75% of flowering plants contain mineral inclusions, calcium oxalate crystals or silica bodies being the most common; other forms are raphides, styloids, druses, prisms or granular sands. Their presence, absence or location in the plants has demonstrated to be of taxonomic relevance. In Orchidaceae for example, raphides and silica bodies are the most common crystals form, when the silica bodies are associated with scelrenchyma fibers they are called stegmata. Stegmata are only known in members of Arecaceae, Marantaceae,
Musaceae, Orchidaceae, Restionaceae, and
Zingiberaceae; particularly in the Orchidaceae they are recorded in four of the six subfamilies. The subtribe
Oncidiinae (subfamily Epidendroideae) includes about
95 genera and 1700 species and is considered on the most derived group in the Orchidaceae. Its members are mostly epiphytes, although several can be terrestrial or lithophytes and distribute from southeastern USA to
Argentina. Based on the assumption that mineral
635

contents can be of potential taxonomic relevance and they can also provide useful characters that support phylogenetic relationships, a study of the mineral inclusions of 140 species of the subtribe was carried out.
The purpose of this study was to describe and compare the mineral inclusions among taxa using as framework a phylogeny recently obtained combining molecular and structural data. In this way we were able to evaluate their role as diagnostic characters and the role they are playing in helping to explain the phylogenetic relationships among Oncidiinae members. Leaf transversal sections were studied with the aid of clear field, phases contrast, and polarized light microscopy. Crystals commonly occurred in mesophyll cells, and only two species recorded them in epidermal cells. Several forms of calcium oxalate crystals and stegmata of silica dioxide deposits were commonly found. On the contrary, crystals like sands or concentric and radial plates were rarely observed; these two latter forms are recorded for the first time in the Orchidaceae. The taxonomic role these mineral inclusions play on the systematics of Oncidiinae is discussed, as well as their value as informative characters based on the phylogenetic relationships of
Oncidiinae members. The importance of continuing with these kind of studies, that complement or enhance results of modern studies, such as molecular ones is stressed.
P0951 – ePoster
Chromosome number in
Solanum sejunctum
: the potential role of polyploidy in the origin of an
Australian dioecious species
, Martine, CT 2 , Anderson, GJ 3
1
Scharf, AR 1
SUNY Plattsburgh, Center for Earth and Environmental
Science, USA; 2
Sciences, USA;
SUNY Plattsburgh, Dept of Biological
3 University of Connecticut, Dept of
Ecology and Evolutionary Biology, USA
The living collection of Solanum sejunctum currently growing in the greenhouse at SUNY Plattsburgh is the first ever and only cultivated population of this species .
In an attempt to better understand the evolution of breeding systems in the genus Solanum , we are researching the pollination biology and origin of divergence between the two sister species ( Solanum sej.
& S. asymetriphyllum ). Solanum sejunctum is a particularly interesting species to study as it is newly identified and was recently found to be dioecious . In all cases within the genus, dioecy is functional; female flowers bear pollen, but that pollen is without germinal apertures (inaperturate) . Solanum sejunctum is geographically limited to a single sandstone outcrop located on Mt. Brockman within the Kakadu National
Park of the Northern Territory of Australia . The two species mentioned above are the only dioecious species found within the Kakadu National Park and are 2 of approximately 15 dioecious species found within the genus Solanum. One way two species can diverge in sympatry is through a polyploid event . David E. Symon
(1981) considers polyploidy a likely generator of diversity in Australian ‘spiny’ solanums ( Solanum subgenus Leptostemonum ) making such a hypothesis plausible for Solanum sejunctum. The chromosome number for S. sejunctum will be identified upon flowering and is part of a larger project to clarify the
636 reproductive biology of this recently described species.
This research has and will continue to expand upon the scientific understanding of a plant genus that is not only economically important but a well-established model system for studying plant evolution.
P0952 – ePoster
Repeated transoceanic long-distance dispersal in the history of the pantrocial genus
Pterocarpus
(Fabaceae: Dalbergieae)
1
Saslis-Lagoudakis, CH 1 , Forest, F 2
Francis, L 2 , Klitgaard, BB 2
University of Reading, UK; 2
, Hawkins, JA 1 ,
Royal Botanic Gardens,
Kew, UK
Traditional views on Gondwanan plant distributions suggest historical vicariance. However, several studies have highlighted the role of long-distance dispersal posterior to the Gondwana break-up in such distributional patterns. Pterocarpus (Fabaceae) is a pantropical genus with its main centre of diversity in tropical Africa (14 taxa), followed by the Neotropics (8 taxa) and Indomalesia (6 taxa). Two species ( P. santalinoides and P. officinalis ) have amphiatlantic distributions between South and/or Central America and western Africa. Previous studies using a family-wide sampling and several fossil calibration points showed that the age of the genus is substantially younger than the
Gondwanan split, refuting Gondwanan vicariance. We generated DNA sequence data from five plastid and nuclear DNA markers for all species of Pterocarpus , analysed them under maximum likelihood and Bayesian criteria and produced a robust ultrametric phylogenetic tree, calibrated using fossil and secondary data. Our results reveal at least four independent events of transoceanic long-distance dispersal in the history of
Pterocarpus . It arose in the Neotropics in the Miocene, dispersed to tropical Africa and subsequently to
Indomalesia. In terms of the amphiatlantic species,
Pterocarpus santalinoides dispersed from South America to western Africa, while P. officinalis crossed the
Atlantic in the opposite direction. Both these dispersal events took place in the Pliocene. Finally, we found that invasions into novel geographic regions are linked with increase in diversification rates.
P0953 – ePoster
Dioecy in the native flora of New Caledonia: incidence, ecological correlates, and origins
Schlessman, M 1 , Vary, LB
1
PP 4
2 , Munzinger, J 3 , Lowry, III,
Vassar College, Poughkeespie, USA;
California Irvine, Irvine, USA;
Noumea, New Caledonia; 4
3
2 University of
IRD, UMR, AMAP,
IRD, UMR, AMAP,
Montpellier, France
The tropical Archipelago of New Caledonia is situated ±
1,200 km E of Australia and ± 1,500 km NNE of New
Zealand. The main island, Grande-Terre, is of
Gondwanan origin, having separated from Australia ca.
85 MYA, although geological evidence indicates a long

submergence until the mid-Eocene ca. 30 MYA . New
Caledonia is the world’s smallest biodiversity hotspot, with over 3,150 indigenous angiosperm species, nearly
80% of which are endemic . Many New Caledonian groups combine disjunct distributions that appear to be
Gondwanan with 'generalized' traits, which led some botanists to characterize them as 'ancient'. More recently, debate has focused on the relative importance of vicariance and long-distance dispersal as explanations for the present distributions of these .ancient Gondwanan' taxa . There is a growing consensus that most, if not all, indigenous New Caledonian lineages have diversified following the mid-Eocene re-emergence . Compared to the floras of other Australasian regions, New Caledonia’s is only moderately known . The first comprehensive checklist was published just 10 years ago, and perhaps
300 species remain to be described . We recently completed the first comprehensive study of the sexual systems of New Caledonia’s indigenous angiosperms.
Because dioecy on islands has been a significant theme in evolutionary botany and since the floras of two other
Pacific Archipelagos, Hawaii and New Zealand, are noted for their high incidences of dioecy, we were especially interested in this sexual system . Through literature, herbarium and field studies, we determined the sexual systems for 98% of New Caledonia’s described indigenous species, finding an unusually high incidence of dioecy: 19.5% . Dioecious species were significantly over-represented among endemics and among taxa occurring solely or primarily in rain forests . Dioecy was also significantly over-represented among taxa with each of three major ecological correlates of dioecy: woodiness, plain flowers, and fleshy fruit . We used the most reliable information available from the published literature to infer both the minimum number of ancestral colonists necessary to account for the extant dioecious flora and the sexual systems and ecological traits of those colonists . We found that, contrary to Baker’s 'law', only
9% of the colonizing ancestors that gave rise to the extant dioecious flora were not themselves dioecious, and that autochthonous derivations of dioecy from other sexual systems have been few and relatively unimportant .
Instead, post-colonization radiations of dioecious lineages have played a significant role, with approximately 90% of the extant dioecious species derived from just 50% of the inferred colonists . We concluded that the high incidence of dioecy on New
Caledonia arose primarily through the establishment and speciation of already-dioecious, woody, rainforest lineages after trans-oceanic dispersal . New Caledonia, with almost 4% of the world’s dioecious angiosperms, mostly endemic, is a rich source of new information on the evolutionary ecology of dioecy . As dioecious taxa are especially vulnerable to the processes that threaten biodiversity, its high incidence on New Caledonia lends additional immediacy to conservation efforts.
P0954 – ePoster
What determines the expression of andromonoecy?
An hypothesis driven comparative study
Schlessman, M 1
1 Vassar College, New York, USA
Andromonoecy is a sexual system in which all individuals produce both perfect (bisexual) and staminate
(male) flowers . In many andromonoecious groups, earlymaturing flowers are perfect, and later-maturing flowers are staminate. This fruit-decreasing pattern is most often interpreted as a response to depletion of resources caused by fruit development . For several reasons, this resource depletion hypothesis is inadequate as a general explanation for andromonoecy . First, such resource depletion must be common in sequentially flowering angiosperms, yet andromonoecy is quite rare . Second, sex allocation theory predicts that the same fruitdecreasing pattern could arise due to floral dichogamy, specifically protandry . Under this dichogamy theory, staminate flowers are favored later in the sequence because later-blooming protandrous flowers are predictably less likely to be adequately pollinated . Third, the dichogamy theory predicts that if perfect flowers are protogynous, staminate flowers will be favored early in the blooming sequence; and this predicted association of floral protogyny and a fruit-increasing pattern of sex expression does occur . Thus, andromonoecy is best understood from the perspective of a more inclusive theory for variation in sex allocation among the flowers of individual plants . Here I report a comparative study of andromonoecious taxa motivated by this more inclusive theory . I predicted that 1) in all taxa, flowers would mature sequentially rather than synchronously; 2) the highest incidence of the expected pattern of sex expression would be in protandrous taxa, where resource depletion and dichogamy would act in concert; and 3) the lowest incidence of the expected pattern of sex expression would be among protogynous taxa, where resource depletion and dichogamy would act in opposition . I have found no evidence contrary to the first prediction . Most of the taxa for which adequate information was available were dichogamous. Seventyfive percent of protandrous taxa had the fruit-decreasing pattern of sex expression, while 57% of protogynous taxa had the fruit-increasing pattern. However, phylogentically explicit tests did not support correlated evolution of the forms of dichogamy and patterns of sex expression . Andromonoecy is seldom characteristic of larger clades, but a notable exception is the
Myodocarpaceae + Apiaceae clade of the Apiales . In that group, andromonoecy appears to be the product of three factors acting in concert: dichogamy, sequentially blooming orders of umbels, and umbels that are compact enough to be a primary functional unit of both pollinator interaction and sex allocation . If umbels, rather than individual flowers, are viewed as the primary reproductive unit, variation in the proportions of perfect and staminate flowers among umbels is analogous to variation in pollen:ovule ratios among individual flowers .
Variation in the proportions of perfect and staminate flowers within umbels produces quantitative differences in sex allocation . Entirely staminate umbels, i.e. zero allocation to ovules, are relatively uncommon . While the presence of perfect and staminate flowers in
Myodocarpaceae + Apiaceae clearly fits the definition of andromonoecy, the quantitative adjustment of sex allocation that it represents is distinctly different from the more qualitative, all-or-nothing abortion of mutliovulate ovaries of the larger flowers of most andromonoecious taxa.
637

P0955 – ePoster
Genetic structure of introduced
Gleditsia triacanthos
(Leguminosae) populations in eastern Australia
Schnabel, A 1 , Chivers, S 2 , Miller, J 2 , Chong, C 3
1
2
Indiana University South Bend, South Bend, USA;
Centre for Plant Biodiversity Research, Canberra,
Australia; 3 Centre for Plant Biodiversity Research,
CSIRO Plant Industry, Canberra, Australia
Honey locust ( Gleditsia triacanthos ; Leguminosae) is an early successional, North American tree species that has been introduced in several temperate regions around the world for urban landscaping and also for agroforestry and livestock fodder in agricultural areas, where it can quickly become naturalized and invasive . Historical records suggest that G. triacanthos was originally introduced to Australia as a fodder tree in Queensland
(QLD), where it quickly expanded its distribution . A concerted eradication program over the past two decades has greatly reduced its distribution and abundance in
QLD . The species, however, has also been introduced for fodder in New South Wales (NSW) and into many cities
(eg, Canberra) as a street tree . We used AFLP markers to study the potential genetic consequences of multiple introductions of G. triacanthos in Australia by comparing population samples from southeastern QLD, eastern NSW, and the Australian Capital Territory
(ACT) . The final data set comprised 139 samples from
11 Australian and 2 North American populations . All samples were scored for 101 AFLP loci . The proportion of common alleles was very high, with only three private alleles detected across all individuals . Estimates of heterozygosity ranged from 0.092 to 0.267, with no significant difference seen between North American and
Australian populations . Australian populations thus appear not to have been subject to any bottleneck effect during and subsequent to introduction from North
America . Significant genetic structuring was found at several levels, with about 71% of the variation found within populations, 16% among populations within regions (US, QLD, ACT, NSW), and 12% among regions. Bayesian clustering analyses suggested that one of the QLD populations was substantially differentiated from the remaining seven QLD populations, but that all
QLD populations show evidence of admixture . The QLD populations were genetically distinct from populations in
ACT and NSW, and the ACT samples grouped more strongly with the US samples than with either QLD or
NSW samples.
P0956 – ePoster
How many species of
Gleditsia
(Leguminosae) are there in southern Asia?
Schnabel, A 1 , Miller, J 2
1 Indiana University South Bend, USA; 2 Centre for Plant
Biodiversity Research, CSIRO Plant Industry, Canberra,
Australia
Gleditsia (Leguminosae) is a genus of 12-14 thorny tree species with a broad distribution that includes eastern
North America, South America, the southern Caucasus,
638 and eastern and southern Asia . The genus shows its greatest species diversity and taxonomic complexity in southern Asia, where species relationships are poorly resolved . Molecular phylogenetic work on the genus to date, which has used sequence data from two chloroplast genes ( ndh F and rpl 16) and the nuclear ribosomal internal transcribed (ITS) spacer regions to compare four of six southern Asian species, suggests that G.
sinensis and G. rolfei form a clade (possibly with G. amorphoides from South America) separate from G. fera and G. australis , which themselves form a clade with the northern Chinese species, G. microphylla. Previous taxonomic treatments had suggested that G. fera is a close relative of, and perhaps conspecific with, either G. sinensis or G. rolfei. In an effort to improve our understanding of the species relationships within southern Asian Gleditsia , we have added collections of the two remaining species ( G. assamica from India and
G. pachycarpa from Vietnam) and increased sampling density of all six species so that we now have two or more accessions of all recognized Gleditsia species in
India, Taiwan, Indonesia, Hong Kong, and Vietnam . We also have acquired several accessions of G. sinensis from
China. Preliminary analyses of DNA sequences from the plastid trn L intron and trn LF spacer regions suggest that
G. fera is, in fact, very closely related to G. sinensis and
G. rolfei. These three species appear to form a clade with
G. assamica and G. pachycarpa , but relationships among those five species are unresolved . The sixth species, G. australis , is clearly differentiated from the other five species and continues to group with G. microphylla .
P0957 – ePoster
On the first tachinid fly (
Diptera
, Tachinidae) carrying Asclepiadoideae pollinaria in the
1
Neotropical region
Nihei, SS 1 , Schwarz, EA 2
Universidade de São Paulo, Brazil; 2 Universidade
Federal do Paraná, Brazil
This is the first record of a Neotropical tachinid on
Asclepiadoideae, only the third world record for
Trichopodini and the first anthophilous record to
Gonolobus parviflorus Decne. (Apocynaceae-
Asclepiadoideae-Asclepiadeae-Gonolobinae). A female of the single species Euacaulona sumichrasti Townsend,
1908 (Diptera-Tachinidae-Phasiinae-Trichopodini) was carrying pol Linaria attached to its proboscis. The new record of the fly specimen was collected from Paraguay,
Depto. Canindeyú, 24°05’S 54°35’W and is deposited at
DZUP. South American plant samples were collected in
Argentina, Bolivia, Ecuador, Peru, Paraguay (eight specimens) and Brazil (less than 20 specimens) and there is no record of it for the locality from which the tachinid fly was captured. Morphometric data and the drawing of one pollinarium were made using optical microscope coupled with a camara lucida. Under natural conditions, there is no evidence that tachinids are able to carry pollinia and transfer them to the flower’s stigma.
Observation of effective pollination is very rare among the Diptera , with only two cases registered so far
(Sciaridae and Chloropidae species on Marsdenia cymulosa in Australia).

P0959 – ePoster
Anther appendages as a taxonomic character in
Cousinia
(Asteraceae)
Sennikov, A 1
1 University of Helsinki, Finland
The shape of apical and basal anther appendages is known to be highly diverse within Cousinia Cass. s.l.
(traditional circumscription of this genus includes also part of Arctium L.). In spite of its diversity, this character has been very seldom used in taxonomic descriptions, delimitation of taxa and comparisons. The apical anther appendages of Cousinia s.l. are lignified (typically of the tribe Cardueae). They are entirely connate before flowering but later split to give way to the growing style.
Our observations from several species native to
Kyrgyzstan (2009–2010) confirmed a very high level of the anther diversity in Cousinia . The shape of the apical part of these appendages is observed to be a stable character, specific to the plants occurring in a certain area and correlating with the other morphological characters, thus being apparently species-specific. In the species studied, the apex of the anthers may be almost abruptly terminated ( C. omissa ), or variously tapering with arched outlines ( C. alpina , C. buphthalmoides , C. fetissowii , C. kasachstanica, C. microcarpa , C. tianschanica ), or gradually narrowed ( C. echinocephala , C. hystricocephala ), or even very finely attenuated ( C. ferghanensis , C. knorringiae , C. simulatrix ). The shape of the apical anther appendages is easily detectable also after the tube has split to release pollen. No visible populational or infraspecific variation has been observed in any species. Instead, the most closely related and difficult-to-distinguish (especially vicarious) species were often found clearly different in the apical anther appendages. In a few cases this character is the only reliable diagnistic feature, complemented with difficultto-grasp characters of pubescence and size of phyllaries and calathidia ( C. echinocephala , C. hystricocephala , C. kasachstanica , C. tianschanica ). The variability in the basal anther appendages is found less prominent, and it is likely related to the section level of classification. The species of C.
section Tianschanicae have the basal appendages shaped as a bunch of threads, composed of a few cells in diameter at their base and branched into unicellular continuations from the middle. The length and diameter of this ‘fringe’ is variable, but at this stage little can be concluded about its significance. Another type of the basal appendages is possessed by the species of C.
section Carduncellus , namely C. alpina , C. buphthalmoides and C. fetissowii , which are broader and have extra small lateral branches of solitary cells at their very base. The species of Cousinia s.l. which phylogenetically belong to Arctium have the apical anther appendages blunt but characteristically acuminate at the apex. The same type of the apical anther appendages is peculiar of Arctium , evidencing about the close relation of this genus and the ‘arctioid group’ of
Cousinia . All in one, the study of anther appendages is very promising to clarify species delimitation and taxonomic relations within the genus Cousinia .
P0960 – ePoster
Environmental factors provide the background for the coherent plant and mammal species distribution patterns in Europe
Heikinheimo, H 1
1
Oikarinen, E 1
, Eronen, J 2
, Uotila, P 3
, Sennikov, A 3 , Preston, C 4
, Mannila, H 1 , Fortelius, M 2
,
3
Dept of Computer and Information Science, Aalto
University; 2 Dept of Geosciences and Geography, and
Institute of Biotechnology, University of Helsinki;
Finnish Museum of Natural History, University of
Helsinki, Finland; 4 Biological Records Centre,
Oxfordshire, UK
Pre-processed presence/absence records of European vascular plants ( Atlas Florae Europaeae database, derived from the published volumes 1–13) and land mammals ( Atlas of European Mammals ) together with basic climate variables (WorldClim database), with a resolution of 50 x 50 km, were used in the analysis of spatial patterns, using cluster and principal component analysis . The principal component analysis suggests that the climatic variables are main factors in forcing plant and mammal cluster distributions. Our analysis shows that vegetation is mainly controlled by temperature, seasonality, and elevation, and that vegetation, to a large extent, guides the mammal distribution patterns. Our interpretation is that PC1 in both mammals and plants is related to species diversity, while PC2 reflects latitudinal gradient patterns, that is, temperature differences and available energy. PC3 appears to reflect seasonality in the form of annual temperature range. The pattern of PC3 for plants is also reasonably similar to topography with a significant correlation with elevation. All of the three first principal components are highly correlated between mammal and plant data. The highest correlation between plant and mammal data is related with mean annual temperature . The clustering of the plant data forms coherent areas that can be interpreted to provide background for floristic regions, reflecting the major factors in the principal component analysis. The first factor affects separation of the largest territories (of the first level splitting, i.e. the highest hierarchy) in Northern
Europe, the Mediterranean region and the British Isles.
This factor is strongly connected with the species number and reflects the diversity and similarity of floras. The second factor is driven by the temperature and affects segregation of the highly oceanic territories in the west and zonal division in the east. The third factor, interpreted as annual temperature range and topography, serves as the basis of the regional clusters which approximately correspond to the floristic regions and their agglomerations . The clustering patterns of mammals and plants form groups that agree with each other in their spatial extent. The forcing of floristic patterns to coherent entities is mainly caused by climatic variables, temperature, temperature range and rainfall, and mediated by elevation differences. The formation of individual plant clusters is also related to species numbers and local and regional floristic differences. The close correlation between floral and faunal patterns suggests that the mammal and plant distributions are controlled by the same environmental variables.
639

P0961 – ePoster
Diversity, differentiation, and vicariance across the
1 species range in the California Sierran endemic plant,
Mimulus laciniatus
Sexton, J 1,2 , Hufford, M 2 , Bateman, A
Meimberg, H 4 , Willis, J 5 , Strauss, S
2 , Lowry, D
2 , Rice, K 2
3 ,
2
University of Melbourne, Parkville, Australia;
University of California, Davis, USA;
Texas, Austin, USA; 4
3 Univeristy of
University of Porto, Vairão,
Portugal; 5 Duke Univeristy, Durham, USA
Species range limits provide rich opportunities to test predictions about the roles of intraspecific genetic variation and gene flow on adaptive evolution. In this study we used microsatellite markers to estimate these parameters in the cut-leaved monkeyflower, Mimulus laciniatus , across an elevational gradient spanning the species range in the California Sierra Nevada . Our main goal was to test established predictions that edges are more isolated and differentiated (‘isolated limits hypothesis’) and to place these findings in the context of theory on constraints to adaptation at range limits . In general, there was no pattern of change in population size, but plant density increased towards margins . No consistent changes in microsatellite diversity was found between the range center and its limits, but there was a reduction in allelic richness in the most peripheral populations, perhaps signaling a step pattern caused by isolation or abrupt environmental changes at range limits .
Mean genetic divergence did not increase significantly towards limits, whereas inbreeding increased gradually across the entire species range, from low to high elevation limits . There was little evidence for reduced gene flow at edges as estimated from graph theory analysis . Gene flow appears to be greater among populations inhabiting similar elevations . Vicariance patterns, potentially driven through environmentally mediated selection, are evident across the elevation gradient, even within the same watershed . Depending on the analysis employed, one can find support for or against the isolated limits hypothesis . Adaptive constraints at range limits in this system may be due to limited genetic variation at both edges, but are not likely to be due to directional, swamping gene flow from central populations.
P0965 – ePoster
Diversity and habitat specialisation among the
Australian samphires (subfamily Salicornioideae,
Chenopodiaceae)
Shepherd, KA 1
Veneklaas, EJ 2
, Moir-Barnetson, L 2 , Colmer, TD 2 ,
1 Western Australian Herbarium, Dept of Environment and Conservation, Australia; 2 School of Plant Biology,
University of Western Australia, Australia
Australia has the highest diversity of stem-succulent samphires in the world. Molecular sequence data suggests that the Australian Salicornioideae colonised
Australia in two separate events. It is likely that the ancestral endemic element (recently included in
640
Tecticornia ) established prior to the arrival of
Sarcocornia , as present day species are more closely related to South African taxa. Limited genetic differentiation among the endemic Australian species suggests that once the ancestral element established the group radiated rapidly. This is concomitant with other studies that reported similar radiations during the late
Miocene to Pliocene among Australian chenopods such as Atriplex and Sclerolaena sens. lat. and a corresponding increase in Chenopodiaceous pollen in the fossil record.
Samphires are 'halophytes' meaning they require salt to thrive and grow and it is likely that the Australian founder element was littoral, facilitating dispersal and establishment along the Australian coastline. The inland arid zone extends to the coast in the northwest of
Western Australia and thus, species that may have established near this region could have readily dispersed inland via the innumerable salt lakes that extend across the continent. While this radiation is reflected in a lack of genetic variation among the endemic Australian
Salicornioideae, significant morphological and physiological variation is evident among species. It is likely that the edaphic heterogeneity of the inland salt lakes, including differences in soil type, salinity levels and the overall size and depth have provided significant habitat variability and facilitated niche specialisation. It is apparent that species exhibit differences in tolerance to the main stress factors in these habitats: salinity, drought and indundation/waterlogging. Consequently, distinct patterns of species zonation can be observed around the margins of ephemeral inland salt lakes .
P0966 – ePoster
Systematic and geography of high mountain flora of the Greater Caucasus
Shetekauri, S 1
1 Tbilisi Botanical Garden and Institute of Botany,
Georgia
Vascular plant diversity of the central and eastern parts of the Greater Caucasus in made up of 950 species, 288 genera and 66 families. The proportion of the leading families (Asteraceae, Poaceae, Rosaceae, Fabaceae,
Caryophyllaceae, Apiaceae, etc.) exceeds 60% of the entire high montane flora. In different habitats (rockscree, subalpine tall herb communities, Caucasian rhododendron thickets, high montane meadows, high montane wetlands, alpine carpets) differ in both species taxonomic and biogeographic structure and coenotic diversity. 69% of the species have Caucasian, Eu-
Caucasian and Colchic types of distribution ranges. The other part is represented by Palearctic, European,
Mediterranean, West Asian and Minor Asian types.
Lithophilous flora is particularly rich in endemic species, as geographic isolation is more pronounced there. In the high montane regions the diversity of micro-habitats is more pronounced in subnival zone. Various rock-screeskeleton substrates such as slate, granite, porphirite, volcanogenic andesite, etc . are wide-spread in the zone.
Structure of the lithological substrates is also different
(fine, moderately fine, coarse, semi-mobile, immobile, mobile scree, rock cracks, etc.). Influence of the
Mediterranean and West Asian floristic centres was significant in the Caucasian florogenesis. Distribution of

Cerastium polymorphum , Saxifraga moschata , S. exerata , Pseudovesicaria digitata , etc. was related with the Caucasian and Mediterranean centres. Colpodium versicolor , Delphinium caucasicum , Minuartia oreina , etc. constitute the Caucasian-West Asian florogenetic group. Alopecurus dasyanthus , Draba supranivalis ,
Noccaea pumila , Pseudobetckea caucasica , Ranunculus tebulosicus , Veronica telefiifolia are isolated in the relevant generic systems and belong to the group of autochthonous species. Comparative systematic– geographic analysis of the Caucasus high montane flora proves that relict plant species of the Tertiary as well as relatively young plants (originated during the glaciation period) participated in florogenesis. Orogenesis and distinct geographic isolation in the Caucasus region was a significant factor in the genesis of allopatric species.
Besides, the large number of species shared by the
Central and Eastern parts of the Greater Caucasus indicates interference of floristic centres of different parts of the Caucasus. Occurrence of such mono- and oligotypic genera on the Greater Caucasus as Eunomia ,
Noccaea , Coluteocarpus , Pseudovesicaria ,
Pseudobetckaea , Charesia , etc. indicates age diversity of the Greater Caucasus high montane flora. Arctic-alpine species are also important in the florogenetic structure of high montane areas, e.g. such species is Saxifraga flagellaris morphologically isolated from the other
Caucasian species of the genus Saxifraga.
P0967 – ePoster
Comparative morphology and structure of colleters of the Rhizophoraceae
Sheue, C-R 1 , Chesson, P 2 , Chen, Y-J 1 , Guu, T-Y 1
, Randrianasolo, RMA 5
, Yong,
, Razanajatovo, JWH 3 , Lim, C-L 4
MH 5 , Yang, Y-P 6
1 Dept of Life Sciences, National Chung Hsing
2 University; Dept of Ecology and Evolutionary Biology,
University of Arizona, USA; 3 Singapore University of
4 Technology and Design, Singapore; Forest Biodiversity
Division, Forest Research Institute Malaysia, Malaysia;
5 Dept of Biology and Ecology, Antananarivo University,
6 Dept of Bioresources, Dayeh University, Madagascar;
Taiwan
The mangrove family Rhizophoraceae has interpetiolar, opposite stipules, with colleters noted in several species.
However, information about colleters for this family is still under-reported, especially for the inland species. In this study, five genera ( Carallia , Cassipourea ,
Gynotroches , Macarisia , Pellacalyx ) of inland species and four genera ( Bruguiera , Ceriops , Kandelia ,
Rhizophora ) of the mangrove species from three tribes
(Macarisieae, Gynotrocheae, and Rhizophoreae) are compared. Two types of stipule were observed. One type is rounded or flattened, with one free margin. The paired stipules of this type overlap each other, and strongly enclose the young shoot. This type is conspicuous (15–80 mm in length) and caducous, and found in both inland
( Carallia , Gynotroches ) and mangrove species. The other type of stipule is small (1–15 mm), more persistent and is without overlapping margins. It is found only in inland species ( Cassipourea , Macarisia , Pellacalyx ). All the species observed have several to hundreds of colleters arranged as rows aggregated in genus specific shapes at the adaxial base of stipules: triangular to semicircular
( Ceriops , Kandelia , Macarisia , Pellacalyx ), rectangular to trapezoidal ( Bruguiera , Carallia ), or as a band
( Carallia , Rhizophora ). The inland genera, Cassipourea and Gynotroches only have the least number (2–10) of colleters arranged in a single row. In addition, several colleters occurred sparsely along the margins of stipules of Pellacalyx , a feature not reported previously. Colleters from members of this family are all anatomically the
‘standard’ type, regardless of whether they appear as sessile rods ( Carallia , Cassipourea , Macarisia ,
Pellacalyx ), stalked rods ( Carallia , Ceriops , Bruguiera ,
Gynotroches , Kandelia ,), or sessile acuminate rods
( Rhizophora ). Colleters in these taxa are constituted by a central axis of slender, elongate cells and an outer palisade-like epidermis perpendicular to the axis, with a secretory function. Based on the size of colleters in this family, three categories of colleters are defined: small (<
300 µm in length), medium (300~700 µm) and big
(700~1200 µm in length). No special association of sizes, shapes, or numbers of colleters with stipule type or with tribe is evident. It is noteworthy that the mangrove species have medium to big colleters, which can be around 10 times larger than those of inland species. The tendency for bigger size, more numerous and compactly arranged colleters of the mangrove species of this family is significant. But the hypothesis that colleters protect young shoots, or have other specific functions, needs to be experimentally verified.
P0968 – ePoster
Chloroplast genome structure of a primitive moss,
Sphagnum palustre
(Bryophyta): DNA sequencing and direct observation of single molecules
Shimamura, M
Deguchi, H 1
1 , Sadamitsu, A 1 , Yamaguchi, T 1 ,
1 Dept of Biol. Sci., Graduate School of Sci., Hiroshima
University, Japan
DNA sequencing and comparison of chloroplast genome structure of bryophytes are expected to provide important information for understanding the evolution of chloroplast genome and early diversification of land plants. We newly provide the chloroplast genome sequence of a peat moss, Sphagnum palustre .
Furthermore, we tested YOYO-1, an intercalating fluorescence dye for DNA, for the visualization of individual chloroplast genome. The chloroplast genome of S. palustre consists of a large single copy (LSC), a small single copy (SSC) and two inverted repeat (IR) regions. Although the gene content is almost identical to bryopsid mosses, the chloroplast genome of S. palustre
(ca. 130 kb) is slightly larger (ca.123.5 kb in Syntrichia ruralis and 122.9 kb in Physcomitrella patens ). In the genome, at least 82 protein, 32 tRNA, 4 rRNA, 4 open reading frames have been detected. Among them, the gene encoding trn P-GGG and ccs A show significant phylogenetic distribution in mosses. These are pseudogenized in bryopsid mosses ( Physcomitrella and
Syntrichia ) but present in Sphagnum . The partial sequences homologous to cys A, cys T, rps 16 and tuf A, which are known from chloroplast genomes of some green algae, liverworts and hornworts as functional genes or pseudogenes, are also detected from S. palustre but
641

these are absent from bryopsid mosses. The phylogenetic distribution of these genes and pseudogenes support the pivotal position of Sphagnum in moss phylogeny. In addition, chloroplasts of S. palustre were purified by percoll density gradient centrifugation, and the DNA fibers were prepared by lyses of intact chloroplasts on the slide glass. After the stainning by YOYO-1, we detected both linear and circular DNA molecules through a fluorescence microscope. Although chloroplast DNA equivalent to single genome (130 kb) are detected as
DNA fibers with 50 µm in length, the molecules of more than 100 µm in length also appear in low frequency.
Larger molecules seem to derive from multimeric conformations of single genomes. In general, chloroplast
DNA is believed to be a closed circular molecule consists of single genome. However, the occurrence of linear and multimeric molecules also have been known in some flowering plants. Such structural plasticity of the chloroplast genome seems to evolutionally conserved across land plants.
P0971 – ePoster
Typification of some species names in
Cordia
(Cordiaceae), with an assessment of the status of
Cordia blanchetii
Silva, T 1 , Stapf, M 2
1
2
Universidade Estadual de Feira de Santana, Brazil;
2Smithsonian Tropical Research Institute, Balboa,
Panamá
Typifications are proposed to facilitate ongoing studies in
Cordiaceae and to maintain current usage. Lectotypes are designated for the following twelve names: Cordia acutifolia, C. salicifolia , C. digyna , C. exaltata var. melanoneura , C. diospyrifolia , C. diospyrifolia , C. sellowiana , C. ulei , C. obscura Cham.
var.
tomentosa ,
Cordia ucayaliensis , C. silvestris , and Patagonula glaziovii . Additionally, nomenclatural notes on Cordia blanchetii are provided.
P0972 – ePoster
Two new species of
Jacquemontia
(Convolvulaceae) from Brazil
Simão-Bianchini, R 1 , Buril, MT 2
1 Instituto de Botânica, São Paulo, Brazil;
Federal de Pernambuco, Brazil
2 Universidade
Jacquemontia Choisy is one of the largest genus of
Convolvulaceae, comprising approximately 120 species, distributed primarily in the Neotropics. The monophyly of the genus has never been tested, but it seems to be a parafiletic group, and some genera have already been segregated from it. The Brazilian species were treated in
Flora Brasiliensis by Meissner in 1869, which referred
33 names and established three sections that are still in use, since that, the species are only treated in local floras or in publishing new species. Recent studies that reviewed the names listed by Meissner, the synonyms and further new species described, cited 54 names that are known to occur in Brazil. The semi-arid region on
Brazil, including vegetation formations as the ‘Caatinga’,
642 savannas and ‘Campos Rupestres’, can be considered one of the diversity centers of the genus, with near to 30% of species and several endemics. Jacquemontia can be characterized by vines or erect subshrub, entire leaves with stellate trichomes, styles single, unbranched, stigmas bilobed with each lobe oval-flat, and the seeds glabrous with two thin wings formed by small trichomes.
Two new species is recognized to the ‘Campos
Rupestres’, both of them are subshrubs, with leaves grey green above (brown in dry), grey beneath, small corollas, pale blue. Jacquemontia sp1 is from Bahia, it resembles
J. decipiens Dammer, easily distinguished by the leaves rounded, with the apex aristulate or apiculate and the sepals unequal, the inner smaller, coriaceous, lanceolated, lanulosous, apex acute with apicule as the leaves, the flowers are densely aggregated into glomerules, at the end of the stems. Jacquemontia sp2 seems like J. ochracea Sim.-Bianch. & Pirani, but the long inflorescence, composed by two cincinnus are very distinctive, the sepals are unequal and rhomboid with the margin undulate and with an acuminate apex, the inner smaller, it was collected in Minas Gerais.
P0974 – ePoster
Molecular phylogeny of
Jacaranda
Juss.
(Bignoniaceae)
Farias-Singer, R 1
Singer, R 2
, Freitas, L 1 , Fregonezi, J 1 , Thode, V 1 ,
1 Depto Genética, Universidade Federal do Rio Grande do Sul, Brazil; 2 Depto Botânica, Universidade Federal do Rio Grande do Sul, Brazil
According to phylogenetic analyses, Jacaranda may be one of the earliest-diverging lineages within
Bignoniaceae. This genus is currently divided into two sections based on the number of thecae: Dilobos, whose anthers display two thecae and Jacaranda (= Monolobos) with just one theca. As a whole, Jacaranda embraces from subshrubs to large trees bearing pinnate or bipinnate leaves. Some species have a broad distribution while others are endemic or restricted to particular regions. The aim of this work was to study the phylogeny of
Jacaranda through molecular tools as well as to infer the evolution of some morphological features within the genus. Twenty eight Jacaranda species representing the morphological diversity within the two sections were sampled. Species of Cybistax , Digomphia ,
Handroanthus , Podranea , Spathodea , Tabebuia , and
Tecoma were used as out-groups. DNA extraction was made from silica-gel dehydrated leaf samples and herbarium vouchers. Sequences were obtained through polymerase chain reaction (PCR) for the following cpDNA genes: rbc L, ndh F and trn Ltrn F. Phylogenetic analyses were performed through Bayesian methods, using MrBayes sofware, with appropriate evolutionary models for each dataset. In order to infer the evolution of some morphological features, character states were plotted onto the tree. Our results suggest that Jacaranda is not monophyletic and that the two sections must be reviewed regarding their delimitation. Digomphia
(originally included as an out-group) is likely inserted within the section Jacaranda . In spite of this, the two sections can be characterized in morphological and geographical terms, as follows: section Jacaranda is

composed by trees bearing bipinnate leaves and monothecate anthers. This section includes two clades, one with exclusive South American distribution, and the other distributed from northern Amazonia to Central
America. Most species within section Dilobos are shrubs bearing pinnate leaves and anthers with two thecae. This section is distributed in central and eastern South
America. Morphological features of Digomphia species
(simple or pinnate leaves, dithecate anthers) are herein interpreted as probable reversals within section
Jacaranda . This study was conducted at the laboratory of
Molecular Evolution, Genetic Department, Universidade
Federal do Rio Grande do Sul. Funding was provided by
CNPq (processo 0151978/208 5).
P0975 – ePoster
The Brazilian Chloraeeae orchids revisited
Singer, R 1 , Buzatto, C 1
1 Depto Botânica, Universidade Federal do Rio Grande do Sul, Brazil
As a whole, orchid tribe Chloraeeae embraces four genera ( Bipinnula , Chloraea , Gavilea and Geoblasta ) which mainly dwell along the Andes, from Tierra del
Fuego (Argentina) to Peru. Remarkably, eight species in this tribe are found well beyond the Andes, at the grassy
Pampa Biome (which comprises Southern Brazil,
Uruguay and part of Eastern Argentina). Six of these species, in turn, are found in Southern Brazil (states of
Rio Grande do Sul, Santa Catarina and Paraná). As a rule, these taxa are seldom collected and, therefore, poorly represented at local herbaria. Literature (either local or international) regarding these species is scant and often misleading. Having all this in mind, we started a research line aimed to elucidate nomenclatural and taxonomical issues within Brazilian Chloraeeae orchids.
In addition, we are documenting biological aspects of these plants (vegetative features, floral morphology, pollination, etc.) which were unknown or poorly known to date. Starting in late 2005, populations have been located during several field trips. Brazilian Chloraeeae orchids are protected by local laws and, therefore, no complete individuals were pressed as vouchers. Only some inflorescences and leaves were pressed. In addition, some leaves were silica-dried for future phylogenetic studies. Floral and vegetative features were recorded in the field, with the help of a digital camera Sony H7.
Pollination studies are being made in the natural populations, with the help of a digital camera and a dcrsr45 Sony video camera as well. In Brazil, there are six
Chloraeeae orchid species: Bipinnula biplumata , B . montana , B . gibertii , B . ctenopetala , Chloraea membranacea and Geoblasta pennicillata . All these species are quite consistent regarding floral features.
With the exception of G. pennicillata , all other species present the labellum articulated at the base of the column.
The anther is incumbent and holds a single pollinarium made up by four yellow, granular pollinia. The pollinarium is devoid of any pollinium stalks and the pollinia adhere onto the pollinator due to a rostellar secretion which is released when the rostellum gets involuntarily pressed by the pollinators. Floral features
(color, indument, etc) in all species except C. membranacea are suggestive of pollination through sexual mimicry. Natural pollination is very good in G. pennicillata and B. gibertii . Pollination in B. montana seems moderated. Ongoing research on the pollination of
C. membranacea points that this orchid is pollinated through a food-fraud system promoted by deceived halictid bees (M. Pedron, pers. comm.). Preliminary phylogenetic studies suggest that Chloraea is not monophyletic, with Gavilea merged within. However, the taxonomic sampling is yet quite incomplete. We hope that the inclusion of the non-Andean Chloraeeae spp . helps to improve the resolution of this orchid group in future phylogenetic analyses.
P0976 – ePoster
Phylogeny of the
Bouteloua curtipendula
complex diploids (Gramineae: Chloridoideae) based on nuclear ribosomal and chloroplast DNA sequences
Siqueiros-Delgado, ME
Columbus, T 3
1 , Ainouche, M 2 , Ainouche, A 2 ,
1
2
Universidad Autónoma de Aguascalientes, Mexico;
Universite de Rennes, Campus Scientifique de Beaulieu,
Francia; 3 Rancho Santa Ana Botanic Garden, USA
The Bouteloua curtipendula complex (BCC) is a group of 12 closely related species within the New World genus
Bouteloua , a member of Poaceae subfamily
Chloridoideae . The BCC is distributed in various ecological settings from southern Canada to Argentina, and its center of diversity is Mexico. The complex is not only highly variable morphologically, but also exhibits a wide range of chromosome numbers (2n = 20 to 103) .
Most species, however, are diploids with 2n = 20 and only Bouteloua curtipendula sensu stricto (2n = 20 to
103), B. warnockii (2n = 20 to 40), and B. purpurea (2n
= 40) are polyploids. Due to considerable morphological variation, putative hybridization, polyploidy and apomixis, circumscription of and relationships among taxa has been uncertain. In order to elucidate the evolution of the BCC, the main goals of this study were to to explore phylogenetic relationships among the BCC diploids, based on nuclear ribosomal and chloroplast sequences. The resulting phylogenetic pattern has been then used to explore the evolution of morphological traits and their usefulness to circumscribe species. Two noncoding DNA regions, ITS and trn T-L-F representing the nuclear and chloroplast genomes respectively, were sequenced and subjected to maximum parsimony analyses. Character state reconstruction analysis was carried out to examine the main morphological characters used in species circumscription. According to these molecular data, the BCC constitutes a monophyletic group. However, the monophyly of the majority of its taxa is not supported, except B. triaena which monophyly is strongly assessed. Although resolution within the BCC was limited with the trn T-L-F data set, the ITS sequences and the combination of the two nuclear and plastid data sets provided a fairly well-resolved phylogenetic pattern of relationships, with the main clades strongly supported.
Present phylogenetic results suggest that the BCC has originally evolved into two main evolutionary lines, which arose from a common diploid ancestor with 2 n =
20 chromosomes, and most likely having yellow anthers and multiple spikelets per branch. Both lineages appear to have mainly diversified in Mexico, with subsequent
643

extension to North and South America. The four character states principally used to circumscribe taxa in the BCC were found clearly homoplasious. The most striking feature arising from our study is that most of the taxonomic units identified at the species level using the traditional morphological diagnostic characters, are not supported by molecular phylogenetic analyses based on
ITS and trn T-L-F data. This provides evidence of a highly artificial classification within the so-called BCC, as it is conspicuously exemplified by the case of B. curtipendula itself and the case of B. media , the two most polymorphic taxa. Circumscription of species in the BCC represents a challenge with regard to the high level of homoplasy found among the traditional diagnostic characters.
P0977 – ePoster
Pollen deposition and seed set in natural populations of
Clarkia tembloriensis
(Onagraceae) which are polymorphic for a homeotic floral trait
Smith-Huerta, N 1 , Huerta, A 1 , Rose, E 1
1 Miami University, USA
Although numerous homeotic mutants have been recognized in laboratory-grown plants, few have been identified and studied in nature . The floral homeotic crinkle petal (cp) mutant of Clarkia occurs in natural populations of C. temboriensis subspecies. tembloriensis and C. exilis. In several populations, cp mutant plants outnumber wild type (WT) plants . This suggests that plants with the cp trait do not suffer reduced fitness, and may in fact benefit from the presence of the trait . The current investigation compares the fitness of cp and WT plants as measured by pollen deposition, pollen tube growth and seed set in two different natural California populations of C. tembloriensis subspecies tembloriensis in 2009 and 2010 . These populations (Crocker Canyon and Elkhorn Grade) consist of plants that are polymorphic for a homeotic mutant of petal development . In the spring, twenty naturally pollinated flowers were collected and preserved in 70% ethanol for each flower type in each population. Twenty additional plants of each flower type in each of the two populations were marked with plastic ties . These same populations were visited again in midsummer and all seed capsules from each marked plant were collected for seed counts .
In the laboratory, the total number of germinated pollen grains secured to the stigma and pollen tubes in the style of preserved flowers was determined with fluorescence microscopy . Seeds in all of the capsules for each plant were counted . Means for pollen grains on the stigma, pollen tubes in the style, and seeds per capsule were determined for each flower type in each population and compared using a Student’s t-test or a Wilcoxon ranked sum test. The flowers of cp plants had more pollen deposited on the stigma, more tubes in the style and greater seed set than WT plants in both populations and both years . These differences were statistically significant with the exception of pollen deposition in the
Crocker Canyon population in 2009, and for seed set in the Elkhorn Grade populations in 2010. The increased pollen deposition, tube growth and seed set in cp mutant plants may help to explain their great abundance in several natural populations of C. tembloriensis. The
644 present study is the first to document differences in the fitness of wild type and homeotic mutant plants growing in natural populations.
P0978 – ePoster
Cytogeography and parallel evolution of rheophyte in
1
Aster ageratoides
in Japan
Soejima, A 1
Kumamoto University, Japan
A rheophyte is a plant that lives along fast moving water currents and sustains irregular flash floods. Rheophytes can survive in such environments because their leaves are streamlined so as put up little resistance to the flow of water. Many plants up to 650 species of 240 genera in 70 families in the world are known to be rheophytes. It is apparently environmental adaptation so that it happens to evolve independently in different river systems in parallel even within a species. Aster ageratoides is one of such species which has rheophyte-like strains in at least four separate areas in Japan. The areas are 1. Shizuoka
Pref., 2. Nara Pref., 3. Yamaguchi Pref., and 4. Miyazaki
Pref. Aster ageratoides itself is distributed throughout these areas and these rheophytes seem to be derived from it independently at each area. To elucidate the paraphyletic origin of these rheophytes and ascertain the presence or absence of gene flow between the rheophytes and its putative ancestors, cytological features including chromosome number and karyotype in addition to morhological features were investigated. It is known that
A. ageratoides is comprised of polyploid series (diploid, tetraploid, and hexaploid) and there occurs taxonomical diversification at each ploidy level. This study revealed that the rheophytes in Yamaguchi and Miyazaki are diploids and those in Shizuoka and Nara are tetraploids.
In Miyazaki, some populations of diploid A. ageratoides are found near the populations of diploid rheophyte, and in Shizuoka and Nara, some populations of tetraploid A. ageratoides are found surrounding the tetraploid rheophytes. It seems that these rheophytes were derived from the ancestors in situ. Although no population of diploid A. ageratoides was found near the diploid rheophyte in Yamaguchi, it is likely that the diploid rheophyte was derived from ancestral diploid near the place, and be a relic after the ancestor extinct (or the diploid ancestor is undiscovered). Based on the morphological investigation, gene flow between rheophyte and its putative ancestor is assumed because there is transitional zone which occupied by morphologically intermediate plants between them.
Therefore, it can be concluded that the rheophytes in A. ageratoides are paraphyletic and still keep genetic contact with their ancestors which makes the taxonomic boundaries unclear.
P0979 – ePoster
Fruit anatomy of
Tragopogon
L. (Scorzonerinae,
Compositae): implications for molecular systematics and taxonomy
Sukhorukov, A 1 , Mavrodiev, E 2 , Soltis, P 2 , Soltis, D 3

1 Dept of Higher Plants, Biological Faculty, Moscow
Lomonosov State University, Moscow, Russia;
Museum of Natural History, University of Florida,
Gainesville, USA; 3
2 Florida
Florida Museum of Natural History,
Dept of Biology, University of Florida, USA
Tragopogon is a large Eurasian genus of approximately
150 species, with a secondary range in North America.
Early systematic studies of the genus were based on fruit morphology. Given recent resolution of phylogenetic relationships in Tragopogon , it is possible to evaluate the evolutionary patterns of these fruit characters .
Approximately 60 species of Tragopogon were included in a new carpological analysis focused on the anatomy of achenes. We found that the following characters are taxonomically significant and may be identified as potential synapomorphies of selected clades of
Tragopogon : (1) the structure of the outer epidermis on the cross section of the pericarp (with or without multilevel cell conglomerations and lignified walls
(“aciculas”)); (2) the structure of unlignified and underlying lignified subzones of pericarp parenchyma;
(3) the vascularization of the unlignified subzone of pericarp parenchyma (the small vascular bundles are present/absent); (4) presence or absence of the hollows in the pericarp; (5) the size of the hollows; (6) the structure of the mechanical tissues; (7) the number of layers in the inner epidermis (one to three); (8) the shape of the crosssection of the body of the outer achene wall; and (9) the shape of the cross-section of the outer achene beak. This new analysis confirms the taxonomical significance of a number of traditionally used carpological characters
(such as the length of the beak, etc.), but, in general, the morphological features of achenes evolved multiple times
P0980 – ePoster
Les Solanaceae du Cambodge, du Laos et du Viêtnam
HUL, S 1
1 Museum National d'Histoire Naturelle, Paris, France
The family Solanaceae comprises about 95 genera and
2500 species, they are cosmopolitan with a high diversity in the western part of South America, where 25 genera are endemic. The genus Solanum , with approximately
1,000 species, is by far the largest of the family. In the
Flora of Cambodia, Laos and Vietnam region, 10 genera and 44 species have been recorded. The family is divided into two subfamilies: (1) The Cestroideae are characterized by usually zygomorphic flowers, bearing 4 stamens, usually capsular fruits (except Cestrum ), seeds oblong or ovoid and a highly variable base chromosome number. They are represented in the region by two introduced genera: Cestrum and Nicotiana , each with one species, and with the majority of their species represented in the New World and Australia, with a single species of Nicotiana present in south-west Africa.
(2) The Solanoideae usually have actinomorphic flowers,
5 stamens, berry-like fruits, discoid seeds, a peripheral embryo and a base chromosome number a multiple of 12.
Two new species are currently in press ( Solanum camranhense Dy Phon & Hul and S. sakhanii Hul) in
Flore du Cambodge, du Laos et du Viêtnam Volume 33, in Faune et Flore Tropicales , IRD-MNHN.
P0982 – ePoster
Integrative taxonomy: a case study in the subgenus
Dermocybe
of
Cortinarius
(Fungi: Agaricomycetidae)
Stefani, FOP 1 , Jones, RH 2 , May, TW 1
1 Royal Botanic Gardens, Melbourne, Australia; 2 Dept of
Primary Industries, Frankston, Australia
The National Herbarium of Victoria houses more than
300 collections belonging to the subgenus Dermocybe of
Cortinarius (Fungi: Agaricomycetidae). Almost 40% of these collections are unidentified to species level.
Phenetic analyses based on macro- and micromorphological characters and pigment chemistry of 110
Australian Dermocybe collections allowed circumscription of 18 species, of which 10 were putatively new. Recently, the concept of DNA barcoding as a tool for species discovery and identification has been introduced. Long before the advent of the barcode concept, the internal transcribed spacers (ITS) flanking the 5.8S ribosomal gene were recognised as one of the best molecular markers to sequence for fungal species identification. Nevertheless fungi remain the only eukaryotes without any official barcoding gene. Here we propose to test the ITS and nuclear large-subunit ribosomal RNA gene as potential barcodes for
Dermocybe species delimitation against the proteincoding genes rpb1, rpb2, tef1 and gpd. Inter- and intraspecific variation and relationships between closely related taxa will be compared across data sets. The congruence between classical taxonomy, chemotaxonomy and DNA taxonomy for the definition of Dermocybe species will be investigated along with the potential application of integrative taxonomy.
P0983 – ePoster
Genetic structure of
Symplocos
(Symplocaceae) in the
Bonin (Ogasawara) Islands using microsatellite markers
Sugai, K 1 , Murakami, N 1 , Kato, H 1
1 Makino Herbarium, Tokyo Metropolitan University,
Japan
The Bonin (Ogasawara) Islands, which are comprised of approximately 20 islands, are located 1,000 km south of
Japan in the northwestern Pacific Ocean. They are one of the oceanic islands and have never been connected to any continental areas. Flora and fauna of such the oceanic islands has been formed by a small numbers of immigrants over the sea. Therefore, genetic variation within a species now growing in the islands is expected very low due to the bottleneck and founder effects.
Consequently, oceanic island’s populations are much more prone to extinction than mainland populations .
Moreover, each island of the Bonin Islands is very small, unlike other famous oceanic islands such as Hawaii and
Galapagos. Population size in the Bonin is inevitably very small, and it made the organisms more feasible to become extinct. The genus Symplocos is monotypic genus in Symplocaceae, comprised of 300-400 species. It is widely distributed in the tropical and temperate regions of East Asia and America. In the Bonin Islands, three
645

species of Symplocos are distributed, and all of them are endemic to the islands. Two of them, Symplocos pergracilis and S. kawakamii are endemic to Chichi-jima
Islands growing in the dry forests and dry dwarf shrubs, respectively. While S. boninensis is endemic to Mukohjima Island of the Haha-jima Islands, which are located
40 km south of the Chichi-jima Islands. All of them are listed as ‘critically endangered’ or ‘endangered’ in the
Japanese Red Data Book from the Environment Agency of Japan. In this study, in order to investigate the detailed genetic structure within the three species of Symplocos in the Bonin Islands, we are trying to determine genotypes of all wild individuals. We call this approach as
‘ubiquitous genotyping’. We tried to collect all wild individuals. At this moment, we could collect 509 individuals of S. pergracilis including two remnant individuals in Ani-jima Island, which is a satellite island of the Chichi-jima Island. We could also collect 14 samples of S. kawakamii, and 142 samples of S. boninensis . We have developed new primers for compound microsatellite markers using the improved technique by Lian et al. (2006). We will report the results on genetic structure within each of the three endemic species of Symplocos in the Bonin Islands.
P0984 – ePoster
Plant diversity and evolution of alpine subnival belt of
Hengduan Mountains, a biodiversity hotspot of SW
China
Sun, H 1 , Li, Z 2 , Xu, B 1
1 Key Laboratory of Biodiversity and Biogeography,
Kunming Institute of Botany, Chinese Academy of
Sciences, China;
University, China
2 Life Science School, Yunnan Normal
Alpine subnival belt is located below the nival line, which is the highest and utmost habitats for seed plants survival, it is an unique and integrity vegetation zone or ecosystem. We investigated the seed plants diversity, evolution and origination from the alpine subnival belt of
Hengduan Mountains SW China by floristic analsying, molecular biogeography, phylgoegraphy from some representative groups. The results showed that there are
846 species, belonging to 135 genera and 43 families, of which 9 genera and 432 species are endemic to
Hengduan Mts and 5 genera, 130 strictly are endemic to alpine scree harsh habitat of Hengduan Mts. Floristic analysis showed that Asteraceae, Saxifragaceae,
Brassicaceae,
Ranunculaceae,
Papaveraceae,
Orobanchaceae,
Caryophyllaceae,
Gentianaceae,
Primulaceae and Ericaceae are at the rank of the fore 10 families of total which containing more than 30 species, these families totally hold 550 species (65.7% total), 65 genera( 48.1% of total), of which Asteraceae is the largest family with 119 species (14./% of total). For the genera, 24 genera of total Saxifraga , Corydalis ,
Saussurea , Pedicularis , Arenaria , Salix , Gentiana ,
Rhododendron , Primula , Cremanthodium , Draba and others include more than 10 species are dominant, of which Saxifraga is the largest with 74 species These genera contain 549 species about 64.9%, of which 318 are endemic (73.6% ) are main composition of the alpine subnival belt flora, many of them formed a differentiate and diversity center in Hengduan Mts. The main floristic
646 elements are N Temperate, Euro–Asia Temperate,
Meditteranean to W Asia and C Asia, C Asia, Sinohimalaya and endemic. Combination with molecular biogeography and molecular lock dating of some representative genera ( Solms-laubachia , Soroseris , Salix etc.) analysis indicated that W. Himalaya or C. Asia (or
Tethys) and Artic-Tertiary are important origination, from which many new taxa origin on the spot during the
Himalaya uplifting and climatic change forming the extreme harsh environment since Pliocene, the rapid diversification and radiation, allopatric speciation and convergent evolution are main evolutionary mechanism.
Phylogegoraphical analysis of some taxa ( Eriophyton wallichii , Thalictrum squaliferum , Syncalanthium souliei etc.) showed that almost population of these taxa in different alpine subnival belt has hardly common haplotype, but has its unique one. The alpine subnival belt of each mountain peak is like a isolated ‘cap’ or
‘island’, which prevent gene introgression in different population among the hill tops or peaks, which accelerate the genetic differentiation of populations and evolution of species in the alpine subnival belt.
P0985 – ePoster
Preliminary molecular phylogeny of the
Daphniphyllum
(Daphniphyllaceae)
Tang, MS 1, 2 , Yang, YP 1 , Sheue, CR 3 , Tsai, CC 4
1 Dept of Biological Sciences, National Sun Yat-Sen
University, Kaoshiung, Taiwan; 2 Dept of Food Nutrition,
Chung Hwa University of Medical Technology, Tainan,
Taiwan; 3 Dept of Life Sciences, National Chung Hsing
University, Taichung, Taiwan; 4 Kaohsiung District
Agricultural Improvement Station, Pingtung, Taiwan
Daphniphyllum , a small-sized genus in the family
Daphniphyllaceae, comprises about thirty species. The phylogenetic relationships of this genus have never been reported. The sectional system of classification of Huang in 1996 was followed. The ingroup contains 12 taxa; 10 species and 2 varieties of Daphniphyllum representing all the three subsections within the genus, and 11 species of outgroups selected for ITS analysis. Phylogenetic relationships were deduced by PAUP* 4.0b10. The amplified ITS regions were 522-658 base pairs (bp) long, among the ingroups. The taxonomic concepts of the
Daphniphyllum are partially incongruent with the results of our phylogenetic analyses of molecular sequence data.
We conclude that Daphniphyllum is monophyletic with the D.
angustifolium and D. buchananiifolium in sister clade of Daphniphyllum for all studied species. Members of the two sections: Lunata and Daphniphyllum , could not form well-supported monophyletic groups, even with
D. buchananiifolium and D. angustifolium placing the basal position. We constructed the first molecular phylogeny of the Daphniphyllum with small subunit cpDNA, and found that taxonomy requires complete revision.
P0986 – ePoster
Homoploid hybrid speciation in Louisiana
Iris
Taylor, S 1 , Martin, N 1

1 Texas State University-San Marcos, USA
Natural hybridization is common in plants and has played an important role in the evolution of plant biodiversity. Although hybridization commonly results in the production of hybrids that are less fit than the parental species, some hybrid lineages may be fit in a novel habitat such that the hybrid lineage can diverge from the progenitor species. Homoploid hybrid speciation involves the evolution of reproductive isolation between such a hybrid lineage and the originally hybridizing taxa . When originally described, Iris nelsonii was hypothesized to be a homoploid hybrid species derived from hybridization between three widespread species of Louisiana Iris (Iridaceae): Iris brevicaulis , I. fulva , and I. hexagona. In order to investigate the hypothesized hybrid origin of I. nelsonii , we first conducted a survey of genetic variation at 15 gene-based and 190 anonymous markers in populations of the three widespread species and I. nelsonii . Second, in order to understand ecological divergence between I. nelsonii and its progenitors, we conducted a common garden experiment to assay the response of the four species to different abiotic conditions that largely characterize their divergent habitats. Results from the molecular work reveal that I. nelsonii shares genetic variation with all three purported progenitor species, supporting the hypothesis that I. nelsonii is indeed a homoploid hybrid species. Furthermore, results from the common garden experiment reveal that I. nelsonii responds to abiotic habitat conditions differently than its progenitors, likely due to adaptation to an extreme deep swamp habitat that is uninhabited by its three progenitor species. Such homoploid hybrid speciation is thought to be a relatively rare event and the Louisiana irises provide a unique system for studying this interesting evolutionary process.
P0987 – ePoster
GBIF + K-samsok = (Almost) True!
Telenius, A 1 , Lundqvist, L 2
1 GBIF-Sweden/Swedish Museum of Natural History,
Sweden; 2 Swedish National Heritage Board, Sweden
GBIF (Global Biodiversity Information Facility) and Ksamsök both represent informatics initiatives aimed at presenting collection data to scientists and to the public.
GBIF mainly serves those interested in natural science, and K-samsök – organizing information on cultural heritage objects – is set up mainly in order to provide the humanities with access to primary and secondary museum data. GBIF-Sweden presently relay information on 27 million objects and observations stored in ca. 100 databases, and K-samsök cover 3.5 million objects from
40 collections nationwide. The cross-over between natural science and cultural interests has been limited, but in trying to establish new ways of looking at old problems, and to find new explanations to old and new issues, a growing body of scientists and lay citizens have found their way by peeping into the opposing field of data. By joining forces GBIF-Sweden and K-samsök will ideally provide access to both groups of high quality data from either web-site: www.gbif.se and www.kringla.nu.
Trials are conducted, and ethical rather than technical objections hinder full-scale operation at present.
Concerns are raised by either side that users will be swamped by ‘irrelevant information’, but by the introduction of more sophisticated search procedures this obstacle will be removed while retaining the possibility of unexpected and thought-provoking experiences…
P0988 – ePoster
GBIF-Sweden 2005–2010: an update on swedish
1 biodiversity collection and observation informatics
Telenius, A 1
GBIF-Sweden/Swedish Museum of Natural History,
Sweden
The objective of GBIF (Global Biodiversity Information
Facility) is to provide free and universal access to the world´s biodiversity data. Almost 270 million objects and observations worldwide of plants, animals and fungi are covered by www.gbif.org, and only in Sweden 27 million data are presented telling about what, where, and when, and by whom an item was collected (some over
250 years old) or an observation made (www.gbif.se).
Starting in 2005 at nil, we are now able to present almost
80 % of the digitized objects present in Swedish natural science collections (where, however, yet another 80% of all items contained await digitization!). In addition to this, old and new observations continue to add to the database, and altogether almost half of all collected and observed primary biodiversity data can be scanned by anyone fitted with a computer and Internet access. Using relevant analytic tools, such information may be valuable to researchers in order to assess the past and present state of species, to model and predict change in relation to varying ambient conditions, and to analyze phylogenetic relationships. Public planning at local to global level is aided given that the whereabouts of species are known, and for educational purpose the availability of this kind of information is crucial in fostering environmental awareness.
P0989 – ePoster
When in doubt, put it in
Phyllanthus
(Phyllanthaceae): towards dismembering a giant genus
Telford, IRH 1 , Pruesapan, K 2 , van Welzen, PC 3 , Bruhl,
1
JJ 1
Botany and NCW Beadle Herbarium, School of
Environmental and Rural Science, University of New
England, Armidale, Australia; 2 Plant Varieties
Protection Division, Dept of Agriculture, Bangkok,
Thailand; 3 Netherlands Centre for Biodiversity Naturalis
(section National Herbarium Nederland), Leiden
University, The Netherlands
Over the last decade there has been a move to grossly broaden the circumscription of Phyllanthus based on preliminary molecular phylogenetic analysis of relatively small samples of segregate genera. Data from floral and fruit morphology and palynology, and phylogenetic trees are presented to indicate that the recognition of 20 or so clades of Phyllanthus sensu lato at generic rank is preferable to the incorporation of genera such as
647

Glochidion , Breynia and Synostemon into a giant genus of more than 1300 species. Diversity within this giant genus would probably result in the naming of the major clades as subgenera, yet these taxa are more likely to be used if presented at generic rank, especially in biogeographical and ecological studies. Many of the names currently used at subgeneric and sectional rank in
Phyllanthus were published as genera prior to Jean
Müller’s broad lumping. Phylogenetic analyses using molecular data show they could be reinstated as monophyletic, morphologically recognisable genera .
P0991 – ePoster
New data for systematic of the genus
Zeravschania
(Umbelliferae) and related taxa based on nuclear
DNA (ITS and ETS) sequences
Terentieva, E 1 , Valiejo-Roman, C 2 , Samigullin, T 2 ,
1
Kljuykov, E 1
2
Botanical Garden, Moscow State University, Russia;
Belozersky Institute of Physico-Chemical Biology,
Moscow State University, Russia
Zeravschania was described by Korovin as a monotypic genus with a type species Z. regeliana distributed in
Central Asia (Korovin, 1948), and further endemic species Z. scabrifolia was found also in Central Asia
(Pimenov, 1983). Five other species ( Z. aucheri , Z. ferulifolia , Z. membranacea and Z. pauciradiata previously treated in Peucedanum and Z. minjanensis denoted in Eleutherospermum ) were added in Flora
Iranica (Pimenov, 1987; Rechinger, 1987). The genus has been recently enlarged further by inclusion Z. stricticaulis and Z. knappii former Peucedanum species
(Pimenov et al., 2007). Morphological and molecular data showed that the closest relatives of Zeravschania are the monotypic genera Demavendia and Haussknechtia
(Pimenov et al., 2005; Valiejo-Roman et al., 2006).
Pimenov (1987) described a new genus Demavendia with only species D. pastinacifolia , distributed mainly in
Western and Central Iran. The genus Haussknechtia was described by Boissier (1872) in his classical Flora
Orientalis based on Carl Haussknecht’s collections of
1868 from Southern Persia (Iran). Thus, this group contains 3 genera Demavendia and Haussknechtia are monotypic and Zeravschania numbers 10 species, but nevertheless the question of the genus volume
Zeravschania remains unclear. Zeravschania is distributed in Iran, Afghanistan and Central Asia
(Pamiro-Alai). In different herbariums (LE, E, MD) there are some specimens similar to the group of three genera listed below, but they were difficult to identify without carpological data. A molecular phylogenetic approach was taken to examine relationships between these genera.
Molecular markers nrDNA (ITS 1,2 and ETS) were used to analyze all species of Zeravschania , Demavendia and
Haussknechtia together with critical samples and their supposed relatives. Parsimony and Bayesian analyses of
ITS 1, 2 and ETS confirmed the close relation of these taxa. All species of Zeravschania , Demavendia and
Haussknechtia together with critical samples formed a well-supported clade. The sample considered as
Demavendia pastinacifolia from East Khorassan (MD) collected in the early stages, has identical ITS + ETS sequences to Z. stricticaulis . The sample from
648
Turkmenistan (LE) is similar to Zeravschania stricticaulis but not identical to it. Specimens from
Western Afghanistan (E) belong to the Zeravschania clade. Thus, the genus Demavendia is distributed only in west and central Iran and is not found in the East
Khorassan. The present preliminary results could stimulate further careful morphological and carpological investigation in order to reveal taxonomic status of critical samples from Turkmenistan (Kopetdag) and
Western Afghanistan (Heart).
P0993 – ePoster
Isolation and characterization of microsatellite markers in the
Barbary thuja
,
Tetraclinis articulata
(Cupressaceae)
Terrab Benjelloun, A 1
Casimiro-Soriguer, R 1
, García-Castaño, JL 1 ,
1 Universidad de Sevilla, Spain
We report microsatellite primer pairs for the Barbary thuja, Tetraclinis articulata (Vahl) Masters
(Cupressaceae). From enriched libraries, we identified more than 200 unique microsatellite loci. We designed primer pairs for 40 of these loci and screened genomic
DNA from 35 to 40 individuals from Spanish, Moroccan and Algerian populations. Eight polymorphic loci were found with a high level of allelic. These highly polymorphic markers will be used in ongoing population genetic studies to evaluate the genetic resources of
Tetraclinis articulata in the W of the Mediterranean.
P0994 – ePoster
Hymenodon
(Orthodontiaceae): exploring evolution and disjunction in a tropical moss genus
Tessler, M 1 , Struwe, L 2
1 New York Botanical Garden and Fordham University,
USA; 2 Rutgers University, New Jersey, USA
A revision of the moss genus Hymenodon
(Orthodontiaceae: Bryophyta) is presented based on morphological and molecular data. Until now six species have been recognized, one of which is composed of three subspecies. Four of the species reside in Australasia
(Malesia, southeastern Australia, Tasmania, New
Zealand, and New Caledonia), while two species are found only in the Neotropics (one in southeastern Brazil and one in the Caribbean). All Hymenodon species grow, almost exclusively, on tree-fern root mantles and may have co-evolved with tree-ferns. Evolutionarily
Hymenodon may also represent the earliest diverging pleurocarpous lineage. The current species concepts are tested using detailed morphological analyses, and evolutionary patterns are evaluated using phylogenetic analyses that incorporate new morphological (30 characters) and molecular ( rps 4 gene and atp Brbc L noncoding spacer) data. Parsimony and maximum likelihood methods were applied for phylogenetic analysis using Leptotheca boliviana , L. gaudichaudii ,
Orthodontium lineare , and Orthodontopsis bardunovii as outgroups. The genus and Hymenodon section
Hymenodon were found to be monophyletic, whereas the

Hymenodon sect. Polystichella was found to be paraphyletic. Because sect. Polystichella was paraphyletic, the sections are not accepted in the new classification. Hymenodon pilifer subsp . sericeus was found as sister to H. chenianus , and not to H. pilifer subsp. pilifer . Accordingly Hymenodon pilifer subsp. sericeus is reclassified here as Hymenodon sericeus .
Biogeographic analysis is also included.
P0995 – ePoster
The Jan and Brigitte Kohlmeyer Herbarium of
Marine Fungi at the New York Botanical Garden
Thiers, B 1 , Halling, Roy 1
1 New York Botanical Garden, USA
In October 2009, Dr. Jan J. Kohlmeyer of the University of North Carolina’s Institute of Marine Sciences donated his herbarium of marine fungi to the Steere Herbarium .
An invaluable collection that includes 6,000 dried and fluid-preserved specimens of marine fungi (on their substrates), 17,500 permanent microscopic slides containing isolated fungal specimens, and associated documentation (field books, notes, photographs, and correspondence), the Kohlmeyer Herbarium has representative material from marine habitats all over the world, including eastern North America, Bermuda, the
Caribbean, Hawaii, Fiji, the Great Barrier Reef, and
Thailand . It is the most authoritative collection in the world of marine fungi, a poorly documented but critical component to the lifecycle of marine ecosystems. In the course of his career, Dr. Jan Kohlmeyer described 149 new species, 50 new genera, four new families, and three new orders of fungi, and produced 158 papers and four books. His book Marine Mycology: The Higher Fungi published with his wife Erika Kohlmeyer, is the definitive reference on this group of organisms. Funded by a grant from the Institute for Museum and Library
Studies (IMLS) the New York Botanical Garden herbarium staff are incorporating the Kohlmeyer herbarium . The overall goals of the project are to: 1) import the Kohlmeyer herbarium database into the
Virtual Herbarium; 2) barcode the 6,000 dried specimens and 17,500 specimens on microscope slides, and add the barcode number to the imported specimen records; 3) interfile the dried specimens; 4) prepare cross-references to be filed in the Steere Herbarium for the material on microscope slides; 5) scan photographic images of specimens; 6) capture digital images of specimens; 7) add images to the database; 8) supplement the specimen data and images with data from field books, photographs, notes, and correspondence; 9) create a web portal to the
Kohlmeyer herbarium; and 10) share these data through the Virtual Herbarium.
P0996 – ePoster
Index Herbariorum, a directory to the world’s herbaria
Thiers, B 1
1 New York Botanical Garden, USA
Currently there are 2695 active herbaria in the world. Of these, 31% are located in Europe and the former U.S.S.R.
(approx. 169 million specimens), 25% are in North
America (approx. 80 million specimens), 23% in Asia
(45 million specimens) 14% are in Latin America (21 million specimens); 5% are in Africa (approx. 9 million specimens), and 2% are in Australasia and Pacifica
(approx. 10 million specimens).Collectively the world’s herbaria contain an estimated 325,000,000 specimens that document the earth’s vegetation for the past 400 years. Information about the world’s herbaria can be found in The New York Botanical Garden’s Index
Herbariorum, available through the garden’s Virtual
Herbarium
(http://sciweb.nybg.org/science2/indexherbariorum.asp) .
The Index Herbariorum (IH) entry for an herbarium includes its physical location, web address, contents (e.g., number and type of specimens), history, and names, birth dates, contact information and areas of expertise of associated staff. Only those collections that are permanent scientific repositories are included in IH. Each institution is assigned a permanent unique identifier in the form of a four to eight letter code, a practice that dates from the founding of IH in 1935 . IH is used to find what herbaria occur in what city, country, or continent; how to contact an herbarium by phone, email or letter; what the strength or focus of the herbarium is in terms of historical plant and fungal groups and geographical areas; what scientists or other staff are associated with an herbarium, and what the research specialties of those staff are . A user can determine when an herbarium was founded, what publications it produces, and what other herbaria it might have subsumed over the years . The IH search page has been accessed more than any other in the science web offerings of NYBG during the entire ten years that the resource has been available on-line. Study of web use statistics and direct feedback from users reveals a wide user base, including other herbaria who use IH as a source of contact information for herbarium commerce; plant and fungal scientists who are conducting biodiversity surveys or evolutionary reconstructions of a taxonomic group use IH to find previously collected specimens that are pertinent to their studies, scientific journals use IH acronyms in the citation of specimens examined for a given study, including voucher specimens in the description of new species; permitting agencies use IH as a resource for determining whether or not an institution should be granted a permit to house endangered species (a CITES permit) and uses IH code as part of the CITES permit designation . An area of future expansion for Index
Herbariorum is support for detailed metadata about holdings in an herbarium, i.e., information on the sources of an institution’s specimen holdings and the taxonomic groups . These data are key to efficient management, including collaborative digitization projects, prioritizing future accessions, and selecting portions of collections for curation or exhibit.
P0997 – ePoster
Resources available for the study of Brazilian plant biodiversity in the New York Botanical Garden's C.
V. Starr Virtual Herbarium
Thiers, B 1 , Kalllunki, J 1 , Thomas, WW 1
649

1f New York Botanical Garden, USA
The NYBG Herbarium has one of the largest and best curated collections of Brazilian plants in the world, with over 500,000 specimens . Brazil, comprising half of
South America, is the most biodiverse country in the world according to the World Conservation Monitoring
Centre . For 75 years, NYBG has been collecting and doing collaborative research throughout the country .
Staff with collecting programs in Amazonian Brazil (ca.
300,000 total specimens) include Boris Krukoff, Ghillean
Prance, Bassett Maguire, Scott Mori, and Douglas Daly.
In central Brazil (ca. 80,000 specimens) staff collectors include Howard Irwin and William Anderson, and in coastal Brazil, especially the state of Bahia (ca. 118,000 specimens); staff collectors include Scott Mori and Wayt
Thomas . For over ten years, NYBG was the coordinating partner in the ambitious Projeto Flora Amazônica and received duplicates of ca. 65,000 collections made during the course of the project . The National Science
Foundation has provided significant support for NYBG's collection efforts in Brazil . Making the wealth of institutional holdings related to the plants of Brazil has been a central objective of NYBG’s institutional digitization plan since the mid 1990s. So far, approximately 400,000 specimens have been at least partially digitized, including all specimens from Brazil’s coastal and central states . Digitization of specimens from
Amazonia is still on-going. Only selected specimens were imaged for specimens from coastal and central
Brazil, but the label data for these specimens were fully transcribed. For Amazonian specimens, all are imaged, but only selected label data are transcribed. Part of the digitization workforce has been 10 Brazilian graduate students, all pursuing Ph.D. degrees at a Brazilian institution . Each Brazilian student worked for approximately six months on the project, during which time they spent half of their time digitizing specimens and the other half conducting their Ph.D. research, using
NYBG facilities and resources . The National Science
Foundation has provided funding for the digitization of
Brazilian specimens at NYBG, including the Brazilian students . Additionally, NYBG is in the process of reimaging all vascular plant types as part of the Global
Plants Initiative, funded by the Andrew W. Mellon
Foundation. This project will result in high-resolution images and full database records for approximately
20,000 types from Brazil . Digitization of systematic literature on Latin American Plants is a focus of NYBG’s
LuEsther T. Mertz Library digitization program.
P0998 – Poster
An investigation of the
Kielmeyera coriacea
(Clusiaceae s.l.) complex using microsatellites
Trad, RJ
V 4 ,
1 , Souza, MBC
Sforça, DA 2
1 , Amaral, MCED
, Zucchi, MI 3 , Souza, AP 2
1 , Bittrich,
, Caddah, M 1
1 Dept of Plant Biology, Institute of Biology, University of
2 Campinas -UNICAMP, Campinas, SP, Brazil; Genetic
Engineering and Molecular Biology Center, Dept of
Genetics and Evolution, Institute of Biology, University of Campinas, Brazil; 3 Agência Paulista de Tecnologia dos Agronegócios, SP, Brazil;
Street, Campinas, SP, Brazil
4 Dr Mário de Nucci
650
The Cerrado, a savanna-like vegetation typical of Central
Brazil, is one of the most species rich biomes of the world. It was classified as a ‘Biodiversity Hotspot’. It is suffering strong anthropogenic pressures of different sources, and quite a number of species are seriously threatened. Kielmeyera Mart. & Zucc. (Clusiaceae s.l.
/Calophyllaceae) is a genus of nearly 50 spp., several of which occur in the cerrado. Morphologically, the genus is characterized by large flowers with a contorted corolla and numerous stamens, a 3-locular ovary, and septicidal woody capsules with numerous winged seeds. One of the most common elements of the cerrado is Kielmeyera coriacea Mart. & Zucc., a morphologically highly variable species. It shows the widest ecological amplitude within the genus and its geographical distribution largely coincides with the cerrado biome.
This species can be easily recognized by the thick corky bark, green coriaceous caducous leaves crown-like condensed at the end the branches, a characteristic venation, and odoriferous, white flowers. In the last revision of the genus Kielmeyera (Nagib Saddi, ined.), the circumscription of K. coriacea remained problematic.
To cope with a high variability, Saddi proposed two subspecies and seven varieties. The species is taxonomically so problematic that it is usually referred to as the ‘ K .
coriacea complex’, to which also belongs K. grandiflora (Wawra) Saddi, formerly included as a variety in K. coriacea . To help with the taxonomic study of this complex, we studied population genetics and structural anatomy of the leaves. Here we present the results of the population genetical study. We used microsatellite due to their well-known capacity to detect polymorphisms at specific level. We selected eleven microsatellite markers previously developed from an enriched library of K. coriacea . K. grandiflora showed a similar banding pattern. For our study we treated each subspecies of K. coriacea as one group and Kielmeyera grandiflora as a third. We sampled at least 5 populations for each group in almost all parts of the cerrado biome: in the states of Minas Gerais, São Paulo, Goiás, Tocantins,
Mato Grosso, Bahia, and in the Distrito Federal. Between
20 and 30 plants were sampled to represent each population. All populations are being genotyped. In this species, the number of alleles per individual ranged from one to eight. The results show how genetically similar individuals of K. coriacea are. Our results do not corroborate most of Saddi’s ideas for a subdivision of the
K. coriacea complex, but suggest that K. grandiflora and
K. coriacea are distinct species (as proposed by Saddi) and that possibly the two subspecies of K. coriacea actually represent two distinct species.
P0999 – ePoster
Molecular phylogeny and biogeography of
Paphiopedilum pfitzer
(Orchidaceae) based on nuclear
1 and plastid DNA
Tsai, CC
HT 1
1 , Chiang, YC 2
, Chou, CH 5
, Huang, SC 3 , Lee, YI 4 , Chuang,
4
Kaohsiung District Agricultural Research and Extension
Station; 2 National Pingtung University of Science and
Technology, Taiwan; 3 Agricultural Research Institute;
5 National Museum of National Science; Research
Center for Biodiversity, China Medical University

The phylogeny and biogeography of the genus
Paphiopedilum were evaluated using phylogenetic trees derived from analysis of nuclear ribosomal ITS sequences, the plastid trn L intron, the trn L-F spacer, and the atp Brbc L spacer. This genus was divided into three subgenera: Parvisepalum , Brachypetalum , and
Paphiopedilum . Evaluation of these sequences together provides higher resolution and better bootstrap support than examination of any of the sequences individually.
Based on the phylogenetic tree derived from the combined data, we demonstrate that each subgenus is monophyletic. Within the subgenus Paphiopedilum , five sections including Coryopedilum , Pardalopetalum ,
Cochlopetalum , Paphiopedilum , and Barbata are divided. The section Coryopedilum was not identified as monophyletic. Instead, it forms a clade with the section
Pardalopetalum . This result suggests that these two sections could be combined. In addition, it was found that the subgenus Parvisepalum also showed a monophyletic character and was located as the basal lineage of this genus. Therefore, we considered the subgenus Parvisepalum as the origin group of genus
Paphiopedilum . The evolutionary trend of genus
Paphiopedilum was deduced based on the most parsimonious tree constructed from the combined sequence data. Based on the observed evolutionary trends, it is likely that Paphiopedilum species were first developed in southern China and were dispersed into
Southeast Asia and the Southeast Asian Archipelagoes.
The subgenera Brachypetalum and Paphiopedilum likely developed after this dispersal event.
P1000 – ePoster
Molecular phylogenetics and ordinal relationships of bryophytes as inferred from a large-scale dataset of chloroplast rbc
L sequences of bryophytes
Tsubota, H 1
Oguri, E 4
, De Luna, E
, Mohamed, H
Seppelt, RD 8 , Yong, KT 9
5
2 , Kubo, H
, Itouga, M
Estebanez, B, Deguchi, H, Itouga, M
1 , Masuzaki, H
, Suleiman, M 6 , Dalton, PJ 7
10 , Shimamura, M,
11 , Shimamura, M
3 ,
,
12 ,
1
Estebanez, B 13 , Deguchi, H 12
Miyajima Natural Botanical Garden, Hiroshima
University, Japan; 2 Red de Biodiversidad y Sistematica,
Instituto de Ecologia, Mexico; 3 Dept of Biofunctional
Science and Technology, Graduate School of Biosphere
Science, Hiroshima University, Japan; 4 Makino
Herbarium, Tokyo Metropolitan University, Japan;
5 Biology Department, Universiti Brunei, Brunei;
6 Development & Human Resources, Institute for Tropical
Biology and Conservation, Universiti Malaysia Sabah,
Malaysia; 7 School of Plant Science, University of
Tasmania, Australia;
Australia; 9 Institute of Biological Sciences, Universiti
Malaya, Malaysia;
8 Australian Antarctic Division,
10 Plant Productivity Systems
Research Group, Plant Science Center, Japan; 11 Plant
Productivity Systems Research Group, Plant Science
Center, RIKEN, Japan; 12 Dept of Biological Science,
Hiroshima University, Japan; 13 Depto de Biologia,
Universidad Autonoma de Madrid, Spain
Bryophytes are the second largest group of modern land plants (embryophytes), after flowering plants
(angiosperms); and they include three major groups: mosses, liverworts and hornworts . Phylogenetic relationships within the bryophytes have been the focus of much attention in recent years . We present the results of a molecular phylogenetic study of the relationships within the bryophytes based on analysis of a large-scale data set of sequences of the chloroplast ribulose 1,5bisphosphate carboxylase/oxygenase large subunit ( rbc L) gene . The results of the analysis suggest that: (1) the liverworts are monophyletic and the most basal lineage within land plants; (2) the Treubiales and Haplomitriales are the most basal lineage within the liverworts; (3) complex thalloids are sister to the members of the other leafy liverworts and simple thalloids as shown in
Masuzaki et al (2010); (4) the mosses are a second basal lineage within the land plants; (5) Sphagnales and
Takakiales are the most basal lineages within mosses; (6)
Andreaeobryales is sister to the remaining mosses and the orders Andreaeales, Tetraphidales, Polytrichales, and the family Oedipodiaceae configure a basal clade within mosses, sister to a large clade with all exemplars of the
Bryopsida as shown in Tsubota et al. (2004); and (7) the hornworts are sister to the vascular plants . The Super-tree constructed using the large dataset supports paraphyly of the bryophytes.
P1001 – ePoster
Detecting initial stage of ecological speciation in the genus
Persea
on the Bonin Islands, Japan
Tsuneki, S 1 , Murakami, N 1 , Yoshida, K 2 , Kato, H 1
1 Makino Herbarium, Tokyo Metropolitan University,
Japan; 2 Faculty of Education and Human Sciences,
National University Corporation Yokohama National
University, Japan
The Bonin (Ogasawara) Islands are located in the western Pacific Ocean about 1000 km south of the
Japanese mainland and are oceanic islands, which have never been connected to any continental areas. Total area of the Bonin Islands is very small (73 km 2 ), unlike other well-known oceanic islands such as Hawaii (1,600 km 2 ) or Galapagos (7,700 km 2 ). However, various types of environments, from dry shrubs to wet forests exist in them. Quick and recurring speciation events adapting such the various habitats, or so-called adaptive radiations might have been occurred frequently there. Many resultant endemic plant species are distributed in the
Bonin Islands. The genus Persea is one of the genera in which such the adaptive radiation occurred in the islands.
This genus has been considered to contain three endemic species in the islands (Toyoda 2003). However, our former studies (Tsuneki et al. unpublished) elucidated that it contains at least six genetically and ecologically differentiated groups. Moreover, three groups out of the six are distributed in one island, Chichi-jima. One group is observed in dry shrubs and the other two are in wet forests there. These three groups are located very close to each other so that considerable amount of gene flow should occur between them. Flowering period of these three groups are overlapped and fertile hybrids were easily produced by our artificial crossing experiments.
Thus, reproductive isolation among the groups has not been established. In the present study, we at first hypothesized that the seeds from each of the three groups are scattered randomly even into different environment from that of their parents because these populations are
651

very close to each other (only 100-300m distant). Then while the offspring are growing, immigrants from different type of the habitat, which are not adapted to the present environment, will die and clearer genetic structure will be gradually formed. To verify this hypothesis, we set up 50m x 50m quadrats in dry and wet environments in Higashidaira area of Chichi-jima, which are only in 100m to 300m distance. We collected leaf samples from all Persea individuals growing in the quadrats including seedlings. We determined genotype of samples using seven SSR markers and classify them into three genetic groups using GeneClass2 (Piry et al. 2004).
Then, we examined relationship between plants size and its genetic groups in different environments. As the plant size become the larger, genetic structure also became clearer, and only genotypes which might be fit to the present environment were observed when they are matured. Thus, we could detect natural selection that reduced genetic variation by removing non-adapted individuals emigrated from different habitats. These results suggested that clear genetic structure, which initiates speciation, could be formed only by habitat differentiation, even without any inner reproductive isolation.
P1002 – ePoster
The New York Botanical Garden's World Flora and
Mycota series: responding to the global strategy for plant conservation's objective of an online Flora of the World by 2020
Tulig, MC 1 , Thiers, BM 1 , Thomas, WW 1
1 New York Botanical Garden, USA
, Mori, SA 1
The New York Botanical Garden began publishing electronic floras and monographs in 2004 in response to the needs of the scientific staff to share taxonomic data immediately, show interim progress in a long-term project, and to instantly provide frequently requested data . Today there are 13 titles in the series that deal with groups of vascular plants, lichens and fungi . The data are all stored in NYBG's Virtual Herbarium database, KE
Software's KE EMu . Unlike more common species pages that aggregate data from across the web based on keyword matches, this publication permits an author to cite the specific references (specimens, images and literature) upon which his/her authoritative taxonomic judgments are based, in keeping with standards of scientific documentation . Treatments typically include the following elements: Names: the accepted scientific name of the species, linked to publication details and to biographical information about the author; a PDF file of the original publication of the species, synonyms for species names and synonym-associated information
(protologs, types, etc.); Descriptions: a species description that includes the morphological features of the species; common names; characterization of the species’ distribution and habitat preferences; data on phenology, pollination and seed dispersal; taxonomic notes including diagnostic features for identification of the species; and a discussion of taxonomic problems, uses, and the meaning of the name of the species;
Distributional information: a distribution map showing known localities where the species is found, generated dynamically from the database records of specimens
652 upon which the treatment was based; Images: illustrating the treatment may be images of the herbarium specimens, photographs of living plants, micrographs, video, and line drawings Links: an unlimited number of links can be made to other on-line content, including specimens of the species in NYBG’s Steere Herbarium and other herbaria
(including images of type specimens), and references consulted in the preparation of the treatment, including gene sequences. Keys to genera and species; are not stored in the EMu database, but are created through interactive identification systems such as LUCID, or as text on a web page (used primarily for dichotomous keys), which are linked to the treatment . The Global
Strategy for Plant Conservation has set as the first target of Objective I: Plant diversity is well understood, documented and recognized as 'An on-line flora of the world by 2020.' The NYBG World Flora and Mycota series provides a model for how this objective might be met . Currently we are in the process of generalizing the tools currently in use so that they can be used by scientists at other institutions to create similar products .
Greater than the technological challenge of creating tools for publishing floristic and monographic data on line is the establishment of criteria for managing an online publication series, including review and editorial oversight and versioning . A challenge yet to be addressed is how to ensure similarly rigorous standards for electronic monographs, whose content may change on a daily basis.
P1003 – ePoster
Lidar and the giants: remote detection of exceptionally tall trees
Tyquin, V 1
1 Forestry Tasmania, Australia
South-eastern Australia is home to the world’s tallest flowering plants, several species in the genus Eucalyptus
(Myrtaceae) . Currently the tallest of these is a 100 m high Eucalyptus regnans growing in the wet forests of southern Tasmania. Eucalyptus regnans is classically the king of the flowering plants, the tallest by nature, but increasingly we are finding other eucalypt species at extreme heights (85 to 100 m), including E. viminalis, E. globulus, E. delegatensis and E. obliqua . Thus, E. regnans may not be the species most capable of attaining extreme height, but rather may be the species most competitive in occupying the environments that allow extreme height growth . Forestry Tasmania manages 1.5 million hectares of multiple-use forest in Tasmania, and has developed policies to conserve giant trees .
Traditionally giant trees have been found during ground surveys, but it is surprisingly easy to miss a giant from the ground. LiDAR (Light Detecting And Ranging technology) is a remote-sensing tool that enables land managers to find stands of giant trees, relate their location to landscape factors, and determine the environmental conditions where extremely tall trees grow. Forestry Tasmania is collecting LiDAR data for the entire area of Tasmanian State forest, to map with great accuracy terrain, slope, aspect, and vegetation layers within forests. Several new stands of giant trees have been detected using LiDAR, including our current highest individual tree, Centurion, measured at 99.6 m by

climbing and tape-drop and at 99.8 m by LiDAR . We will use this information to determine the frequency of giant trees across the environmental range of species known to produce giants, and the environmental conditions that foster extreme height growth . This will allow forest management planning for conservation of future giants, especially with consideration of climate change effects .
P1006 – ePoster
The genus
Aulonemia
(Poaceae: Bambusoideae) in
Brazil
Viana, PL 1 , FIlgueiras, TS 2 , Paiva, EAS 3
1 Jardim Botânico Inhotim, Brumadinho, Brazil; 2 Instituto de Botânica de São Paulo, São Paulo, Brazil;
3 Universidade Federal de Minas Gerais, Belo Horizonte,
Brazil
Aulonemia Goudot is a genus of woody bamboo (tribe
Bambuseae) with 40 known species. It belongs to
Arthrostylidiinae clade, one of the three subtribes of woody bamboo in the Neotropics. Together with
Chusquea and Merostachys , Aulonemia is one of the three more speciose Neotropical bamboo genera. It is distributed from Mexico to Southern Brazil, where the species occur usually in high altitude sites, such as the
Andean páramos, the tepuys in the Guyana shield and high altitude forests in Central America and Brazil. In
Brazil, almost all the Aulonemia species are endemic to the threatened Atlantic Rain Forest. As part of an ongoing collaborative taxonomic revision of the genus, a taxonomic study of the Brazilian species of Aulonemia was carried out during last four years. Intensive fieldwork throughout Brazil was performed in order to supply the lack of bamboo specimens in most herbaria, especially in Brazil. Additionally, the main herbaria in
Brazil and United States were studied, as well as type specimens from European herbaria. Detailed morphological descriptions, keys for identification of the species, maps of geographic distribution and ecological and conservation data were provided. Leaf blade anatomy of the species was assessed and evaluated as a taxonomic tool for Aulonemia. The anatomical studies included cross sections of the leaf blade and investigation of the epidermis under a scanning electron microscope.
Maps of geographic distribution of the species were made and their conservation status was evaluated according to IUCN criteria. A total of 14 species were confirmed to occur in Brazil: A. amplissima, A. aristulata, A. deflexa, A. effusa, A. goyazensis, A. lanciflora, A. radiata, A. setigera, A. setosa, plus five new species, which are also presented here. Four new synonyms and four lectotypes are proposed. Half of the inventoried species were considered under threaten of extinction, four of these critically threatened, two endangered and one species vulnerable. Aulonemia goyazensis is probably extinct, because it has not been found in nature after intensive search. Leaf blade anatomy characters considered useful for delimiting species or group of species of Aulonemia include distribution of intercostal sclerenchyma, presence/absence of fusoid cells in the mesophyll, type of vascular system in the midrib, distribution of stomata, and distribution and characterization of epidermal appendages like prickles and papillae.
P1008 – ePoster
Evolution and biogeography of
Pleurophyllum
(Asteraceae), a small genus of megaherbs endemic to the Subantarctic Islands
Wagstaff, SJ 1 , Breitwieser, I 1 , Ito, M 2
1 Allan Herbarium, Landcare Research, Lincoln, New
Zealand; 2 Dept of Systems Sciences (Biology), University of Tokyo, Komaba, Japan
The abundance of fossils in Antarctica suggests this continent was a center of plant diversification and may have served as a corridor for migration for many austral plant groups until the late Cenozoic. While the Antarctic flora was largely extirpated by the Miocene, we test the hypothesis that at least some Antarctic plant lineages may still persist on the Subantarctic Islands and have dispersed in a northerly direction to New Zealand. We used independent and combined analysis of ITS, ETS, trn K and trn L DNA sequences to infer the origin and patterns of diversification in Pleurophyllum , a small genus of three species that are endemic to the
Subantarctic Islands of Australia and New Zealand.
Divergence times were estimated using a Bayesian framework with recent fossil evidence for Astereaceae as well as the emergence of the Chatham Islands as maximum and minimum calibration points. Error associated with these calibrations was incorporated using probability distributions. In our analysis Damnamenia emerged as sister to Pleurophyllum and a group of macrocephalous Olearias that are endemic to New
Zealand. High posterior probability values suggest this lineage originated in the Subantarctic islands then diversified in a northerly direction to mainland New
Zealand and the Chatham Islands. The inferred stem age of 10.7 (8.5–21.9) Ma for this lineage largely predates
Miocene extinctions in Antarctica, hence the ancestor of these plants could have been more widely distributed on the Antarctic continent. However the crown radiation occurred at the boundary between the Miocene and
Pliocene approximately 5.1 (2.2–9.5) Ma corresponding closely with episodes of glaciation in New Zealand.
Pollen resembling Pleurophyllum is found in abundance in the oldest peat deposits on Campbell Island that were deposited close to the Last Glacial Maximum approximately 13,000 years ago.
P1009 – ePoster
Species-specific identification of
Larix gmelini
,
L. olgensis
and
L. principis-rupprechtii
by SCAR marker technology
Wang, Q 1 , Qu, L 1 , Liu, D 2
1 Northeast Forestry University;
Technology, PR China
2 Harbin Institute of
Larix gmelini, L. olgensis and L. principis-rupprechtii are three kinds of important Larch species in forestation in North China, with different growth characteristics and different distribution range. It was very difficult to
653

distinct the seeds of them visually in the seed's purchasing, managing and allocating because of the extremely similarity in seed outline. It would cause heavy economic lose if seeds were misused in forestation. In this study, each species sample group was composed of 50 randomly selected seeds as the materials, the randomly amplified polymorphic DNA (RAPD) fingerprint technique was used as a tool for typing the these Larch species and to find specific markers for evaluating their genetic differentiation. 47 random RAPD primers were used to amplify the haploid genomic DNA from their seed endosperm. The species-specific DNA segments of these three Larch were extraction from agarose gel and sequenced, and SCAR primers were designed according to the 5' and 3' end sequences. The unique species-specific segments which were amplified by the SCAR primers were obtained which provided accurate, convenient SCAR marker for the identification of larch seeds. The results indicated that significant species-specific bands amplified on Larch haploid genomic DNA by the SCAR molecular marker designed from RAPD segments could be obviously detected. It could be concluded that there was significant genetic variation among these three larch species in the genomic
DNA level and SCAR molecular marker technique could be used to identify these Larch seeds in its advantage of good stability, high repeatability, easy operation and short test cycle. It would be actually feasible to identify the seeds of Larix gmelini, L. olgensis and L.
principisrupprechtii by SCAR marker as long as the proper primers and appropriate number of samples were decided. The problems of forestry such as the larch seed identification, deployment of larch seeds and planted at suitable location etc. could be well solved by SCAR markers obtained in this study. It could also provide a scientific identification method to avoid economic loss by misuse of larch seeds. The result also showed that
RAPD and SCAR primers can highly repeated in the bands of species-specific fragments.The percentage of fragment OPB-111500 amplified by SCAR primer
SCB1/SCB2 was 92% in seed lot of Larix gmelini and
90% in seed lot of Larix olgensis . The OPN-17400 fragment amplified by SCN1/SCN2 was presented 86% in L.principis-rupprechtii seed samples and the OPX-
141200 fragment amplified by SCX1/SCX2 was presented 88% in Larix gmelini seed samples. The percentage of species-specific fragments amplified by three SCAR primers was all higher than that of RAPD primers. It was declared that the specific SCAR primers could be used as a reliable molecular technique to identify the seeds of different larch.
P1010 – ePoster
Phylogeny and evolutionary history of the large genus
Dyckia
(Pitcairnioideae; Bromeliaceae) in eastern
South America, inferred from chloroplast and nuclear sequence data
, Weising, K 1
1
Krapp, F 1
Institute of Biology, University of Kassel, Germany
The genus Dyckia (Pitcairnioideae, Bromeliaceae) consists of ~140 xerophytic species that are distributed in azonal habitats of Neotropic savannas and the Atlantic forests of Brazil and adjacent countries. Dyckia shows an
654 enormous species richness, as compared with the closely related genera Encholirium (23 species), Deuterocohnia
(18 species), and Fosterella (31 species). Up till now only few Dyckia species have been included in molecular systematic investigations. Consequently, little is known yet about infrageneric relationships and the biogeographic events that generated the present-day distribution patterns. Not even the relationships between
Dyckia and its putative sister genus Encholirium have been unambiguously resolved. Here we present a phylogeny of Dyckia and related genera, based on six chloroplast DNA loci and the nuclear phytochrome C gene (PHYC). Sequence variation within Dyckia turned out to be extremely low, as was also reflected in the low resolution of the trees. Nevertheless, several major clades could be identified that roughly correspond to the geographic origins of the samples. Whereas Dyckia is apparently monophyletic, the status of Encholirium is still ambiguous due to limited sampling. The dated chloroplast phylogeny provides evidence that the genus experienced a very recent radiation, except for two early branching lineages that show overlapping distribution areas with Encholirium in eastern Brazil. Our current data are compatible with the hypothesis that the ancestors of Dyckia and Encholirium have existed in eastern Brazil for quite some time before one lineage became capable of spreading into the today-distribution area of Dyckia .
Identifying possible adaptations associated with that spread and the rapid diversification that followed could provide a model for radiations in eastern South America in general.
P1011 – ePoster
Evolution in the Central Andes: phylogeny and phylogeography of
Fosterella
(Pitcairnioideae,
Bromeliaceae)
Wagner, N 1
Schulte, K 3
, Michalak, I
, Weising, K 1
2 , Silvestro, D 2 , Zizka, G 2 ,
2
1 Institute of Biology, University of Kassel, Germany;
Botany and Molecular Evolution, Research Institute
Senckenberg and Goethe-University, Frankfurt,
Germany; 3 Australian Tropical Herbarium, James Cook
University, Cairns, Australia
The Neotropical genus Fosterella (Pitcairnioideae,
Bromeliaceae) includes 31 species with a centre of distribution and diversity in the Bolivian Andes. Most of the species are characterized by small ranges and can be found from lowland Amazon forest and isolated mountain ranges of the Brazilian shield to cool cloud forest and high arid valleys in the Andes. In our ongoing project we combine molecular and biogeographical data to get insights into the evolution of Fosterella , its origin, expansion and the formation of endemic species.
Comparative sequencing of six chloroplast DNA regions
( mat K, rps 16 intron, atp Brbc L, psb Bpsb H, rpl 32trn L, rps 16trn K) resulted in a well-resolved phylogeny.
Monophyletic Fosterella was divided into into six distinct evolutionary lineages. The same general tree topologies were obtained by sequencing portions of the nuclear PHYC and PRK genes. Applying a molecular clock to the chloroplast tree resulted in an approximate age of the stem group of Fosterella of 10 My, coinciding with the onset of a major uplifting phase of the central

Andes. We applied AFLPs to increase the phylogenetic and phylogeographic resolution, and to study speciation processes within the subgroups defined by the chloroplast data. AFLP results are presented for the F.
rusbyi group and the F. penduliflora -group. The latter is particularly interesting because it comprises the morphological diverse and widespread species F. penduliflora and the narrow endemic F. gracilis in sister positions.
P1012 – ePoster
Biogeographic analyses of Araliaceae in Asia
Wen, J 1 , Li, R 2
1
2
Smithsonian Institution, Washington DC, USA;
Kunming Institute of Botany, China
Asia is a major center of diversity of Araliaceae . We constructed the phylogeny of two major lineages of
Araliaceae in Asia: the Hedereae (20 genera and ca. 650 spp., with 18 genera and 201 species sampled) and the
Aralieae (two genera: Aralia L. and Panax L. and 90 species, with both genera and 70 species sampled) using seven molecular markers (the nuclear ribosomal ITS and the plastid ndh F gene, the trn Ltrn F region, the rps 16 intron, the atp Brbc L intergenic spacer, the rpl 16 intron, and the psb Atrn H intergenic spacer) . Phylogenetic analyses of the combined plastid and ITS data suggested the monophyly of both tribes. The reconstructions of historical biogeography were conducted on the maximum clade credibility (MCC) phylogeny of the combined data set . The ancestral area distribution of both Aralieae and
Hederieae was inferred using the newly developed maximum likelihood-based method Lagrange . Five areas of endemism were defined based on the taxon distributions and our interests in Asian biogeography: eastern Asia, subtropical and tropical continental Asia, the Malesian region, Europe, and the New World . Based on biogeographic analyses and fossil-calibrated Bayesian dating, the Hedereae is suggested to have originated in eastern Asia in the late Cretaceous around 72 mya . The ancestral area of Aralieae is inferred to be in the New
World and taxa of the tribe migrated into eastern Asia via two events . The Malesian taxa of both Hedereae and
Aralieae were derived from continental Asian relatives, and subsequently radiated throughout the Malesian region primarily in late Miocene and Pliocene . Southern
China and Indochina were critical in the early diversification of three major species-rich tropical genera: Dendropanax, Trevesia and Schefflera of the
Hedereae in the Eocene and Oligocene . Eastern Asia was a major diversification center of Aralieae in the Miocene, corresponding to the active orogenies in continental Asia due to the uplift of the Himalaya and the subsequent climatic changes in the region in the late Tertiary . The
Gondwana seems important for the early evolution of the family Araliaceae; yet our results suggest the importance of eastern Asia in the early evolution of the species-rich and primarily tropical Hedereae.
P1013 – ePoster
The root of the dictyostelid tree
Weststrand, S 1 , Baldauf, S 1
1 Systematic Biology, Uppsala University, Sweden
The root represents the oldest point in a phylogeny and determining it gives evolution a time arrow. The first molecular treatment of the social amoebae (Dictyostelia) was carried out with small subunit ribosomal DNA (SSU rDNA) data in 2006, and from this four major dictyostelid groups were defined. However, the relationships among them, i.e. the root of the tree, could not be confidently determined. In this study, a new protein data set – eukaryotic release factor 3 (eRF3) - was developed for deep dictyostelid phylogeny. This was accomplished by designing degenerate PCR primers, developing PCR strategies, generating sequences from seven new species and assembling currently available dictyostelid data sets. Phylogenetic reconstruction according to a strict total evidence approach supports the earlier SSU rDNA root, placing group 1 as the most basal divergence. However, an alternative root lying between group 1 + 2 and group 3 + 4 has substantial support, especially from protein coding sequences. In addition, the analyses confidently place Dictyostelium purpureum as the earliest branch in group 4. This result questions the age of group 4 and the calibration of the whole dictyostelid phylogeny.
P1014 – ePoster
1
The mating habits of
Chiloglottis
orchids
Whitehead, M 1 , Peakall, R 1
Research School of Biology, Australian National
University, Canberra, Australia
The terrestrial orchid genus Chiloglottis secures pollination by sexual deception whereby male insect pollinators are attracted to the plant by mimicry of specific sexual signals. Remarkably, sexual deception has evolved independently on at least four continents, with a wide range of insects known to be exploited. The
‘outcrossing hypothesis’ posits that by hijacking the sexual behaviour of their pollinator, sexually-deceptive orchids ensure their pollen is transported far enough away from the parent plant to promote outcrossing.
Furthermore, divergent floral volatile chemistry linked to different pollinator species is predicted to act as a strong pre-zygotic reproductive barrier, thereby limiting hybridization in sympatric species . Investigating pollen mediated gene flow within and between taxa is usually the target of mating system analysis, however despite the bank of literature on sexually deceptive orchids and orchid pollination more generally, mating system analyses are rare for orchid species. This is largely because of the unique challenges posed by orchid seed which is miniscule and requires infection by a specific mychorrizal partner for germination . This study addresses these challenges through symbiotic germination of orchid seed collected from naturally pollinated orchid pods. Outcrossing measures are obtained by genotyping orchid offspring at six microsatellite loci and show that Chiloglottis orchids occupy an extreme position on the spectrum of plant outcrossing rates. Measures of neutral genetic variation within and between sympatric orchid taxa suggest that interspecific gene flow is absent or exceedingly rare.
Along with data from pollinator choice and mark-
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recapture experiments the results indicate that sexual deception is finely adapted to maximise outcrossing despite clonality and that divergent floral odour is capable of maintaining strict pre-zygotic reproductive isolation in sympatric taxa.
P1015 – ePoster
Emonocot: biodiversity informatics for monocots and monocot systematists
Wilkin, P
HCJ 2
1
MJ 3
1 , Barker, A
, Jackson, M 1
, Simpson, DA 1
1 , Baker, WJ 1
, Kitching, IJ 1
, Smith, VS 3
, Clark, B 1
, Mayo, SJ
, Godfray,
1 , Scoble,
Royal Botanic Gardens, Kew, UK;
Tinbergen Building, Oxford; 3
2 Dept of Zoology,
Dept of Entomology,
Natural History Museum, London
The eMonocot project which started in late 2010, aims to change the methods by which biodiversity information is delivered to its users as taxonomy develops into an eScience. The project is creating an authoritative webbased treatment of monocot plants by generating ~70,000 outline species pages based on the World Checklist of
Monocotyledons
(www.apps.kew.org/wcsp/monocots/home.do). These will be supported by keys and taxon pages to all the families of monocots and ~2000 genera in 8 major families. Comprehensive species pages will include
European monocots (~2000 species), 'Sampled Red List
Index' monocots (1500 species) and Cypripedioideae
(slipper orchids) (~130 species) with over 20,000 species having enhanced content. Further taxa will be delivered via 'Scratchpads' (http://scratchpads.eu), community websites for taxonomists geared to uploading and presenting a wide variety of taxonomic data. Existing monocot web resources such as CATE-Araceae
(www.cate-araceae.org), Palmweb (www.palmweb.org) and Grass Base (www.kew.org/data /grasses-db.html) represent important building blocks around which the eMonocot system will be built. The project will be linked to other resources maintained by key international stakeholders in biodiversity informatics. eMonocot will generate software tools to permit future taxonomic research to be web-based, enabling monocot scientists anywhere in the world to participate, creating social structures and working practices to facilitate and manage these global interactions. This global participation will be critical in developing and sustaining both the data of eMonocot and the communities that will generate and enhance it .
P1016 – ePoster
The ecology, biogeography and systematics of the
Austro-Papua
Nepenthes
Wilson, G 1, 2
1
2
Australian Tropical Herbarium, Cairns, Australia;
School of Marine & Tropical Biology, James Cook
University, Cairns, Australia
Little is known of Nepenthes in the Austro-Papuan region and there is conjecture about the number and status taxa present . In Australia as many as 11, few as
656 one, and currently three, species of Nepenthes have been recognised as occurring. Nepenthes mirabilis (Lour.)
Druce is the most widespread, from the Wet Tropics through to southern China, overlapping all except six of the c. 125 described species. The others, Nepenthes rowaniae F.M.Bailey and N. tenax Clarke & Kruger are known only from swamps north of the Jardine River on
Cape York, Queensland and not from New Guinea despite its proximity and similarity of habitat. This raises the question of where and when they evolved and if they have been overlooked on New Guinea. In addition (1) several populations of Nepenthes of uncertain taxonomic status occur in Queensland (see Clark & Kruger 2005,
Beasley 2009) and warrant further investigation, and (2) collections from Cape York currently in the National
Herbarium, Canberra, are identified as N. alata , otherwise known only from the Philippines, and N.
alicae F.M.Bailey, but reduced to a synonym of N. mirabilis by Danser (1928), and their identity requires clarification, and (3) we have no knowledge of the ecology of any species in the study area . Studying
Nepenthes in Australia is often challenging, e.g. N. tenax and N. mirabilis occur in swamps and many plants are submerged in the wet season and subjected to wildfires in the dry season. It is further complicated as the species hybridize and undescribed entities also occur. One, colloquially known as ‘mini-tenax’, occurs far into the swamps and is proving particularly difficult to study; crocodiles have little respect for botanists loitering in waist-deep water or pre-packed in aluminium containers .
To define species, describe the ecology, understand the evolution, and determine the status of Austro-Papuan taxa and contribute to a wider study of Nepenthes being coordinated by Dr Charles Clarke in Kuala Lumpur we are collecting morphological, morphometric, ecological and gene sequence data. Duplicates of collections are being deposited in state and national herbaria and sequence data in Genbank. In 2010 we sampled populations of three species and several indet taxa across a range of habitats, re-sampled a population of dubious taxonomic status in Queensland, and commenced ecological studies. Planning is in train for the 2011 field season in Cape York and 2012 in Papua New Guinea and
West Papua and publication of results.
P1017 – ePoster
Analysis of the DNA barcode marker variation of
Salix
species in South Korea
Jin, BB 1 , Lim, CK 1 , Won, H 1
1 Daegu University, Republic of Korea
Short flowering period separated with long vegetative growing season, dioecy, frequent hybridization and lack of identifiable characters, etcs hinder correct identification of Salix species. To correctly identify Salix species, those DNA barcode markers such as chloroplast trn L intron, trn Ltrn F IGS, psb Kpsb I IGS, psb Atrn H
IGS, rpo C1 gene, rpo B gene, and trn K 5'mat K genetrn K 3' region were PCR amplified and sequenced, in addition to nuclear single copy xdh gene marker.
Sequence variations were analyzed and clustering by parsimony analysis were performed to test which DNA barcode is best in identification of Salix species. As results, psb Atrn H IGS and trn K 5'mat K genetrn K

3'region was most successful in recognizing 11 groups from 19 taxa when markers were used alone. When two markers were analyzed together, three combinations – trn L intron + psb Atrn H IGS, psbKpsb I IGS + psb Atrn H IGS, psb Atrn H IGS + trn K 5'mat K genetrn K 3' – showed best resolution that recognized 15 groups out of
19 taxa. Maximum resolution was achieved when three markers were jointly analyzed. Two combinations – trn L intron + trn Ltrn F IGS + psb Atrn H IGS, trn Ltrn F IGS
+ psb Atrn H IGS + trn K 5'mat K genetrn K
3'combination – recognized 18 groups. However, nuclear xdh gene sequence alone recognized 19 groups which corresponds to or surpass the resolution of the three marker combinations of the chloroplast DNA barcode markers. As two accessions of S. chaenomeloides showed infraspecific variation, nuclear xdh gene outnumbered the result of chloroplast DNA barcode markers where the two accessions have identical sequences. These results indicate that chloroplast noncoding markers are more economic and effective than coding markers and nuclear xdh gene surpass those chloroplst markers. Nonetheless, markers employed in this study were not enough to clearly resolve the phylogenetic relationships among the Salix species.
Therefore we need fast evolving nuclear markers that have more informative contents than xdh genes to reliably resolve species level relationships among Salix species.
P1018 – ePoster
Teaching of botany the miscellaneous way
Woodland, DW 1
1 Dept of Biology, Andrews University, USA
The teaching of botany and especially organismal plant biology, has become less and less in many universities in
Australia, Europe, New Zealand and North America .
Many institutions no longer offer any nonphysiological/molecular botany courses, especially courses in plant biodiversity, systematics, floristics or field botany to give students an outdoor experience . The purpose of this display is to show how educators can stimulate interest in the botanical world by using other avenues of learning, different from traditional science.
This can include history of biology and classification of the plant kingdom using images from postage stamps, architecture, signage, pop culture, etc. Plant biology can also be taught by exploring basic concepts using music, literature, movies and the virtual world. Many examples illustrating these ways will be provided to stimulate student learning.
P1019 – ePoster
A biogeographical contact zone of
Euphrasia
(Orobanchaceae) inferred from molecular studies
Huang, S-F 1 , Lin, Y-H 2 , Huang, T-C 3 , Wu, M-J 2
1 Dept of Applied Science, National Hsinchu University of
Education, Taiwan; 2 Dept of Natural Resources and
Environmental Studies, National Dong Hwa University,
Taiwan; 3 Institute of Plant Biology, National Taiwan
University, Taiwan
Euphrasia is a genus of bipolar distribution connected by tropical and subtropical mountain peaks in Taiwan, the
Philippines, Borneo, Sulawesi, Seram and New Guinea.
Fifteen sections are recognized and two sections,
Malesianae and Pauciflorae , are restricted to the connecting areas. Both Southern Hemisphere and
Northern Hemisphere origins have been proposed.
Recently, origin in the Northern Hemisphere was supported by Gussarova and her colleagues based on a phylogenetic tree of Euphrasia reconstructed by nuclear
ITS and chloroplast DNA markers including atp Brbc L, trn L intron and trn Ltrn F intergenic spacer with worldwide samples excluding the section Lasiantherae from Australia. This study also supported the view that
Euphrasia has migrated from north to south through the connecting areas. At the same time, we published a biogeographical study of Euphrasia emphasizing sections of Malesianae and Pauciflorae in the connecting areas including species from Taiwan, the Philippines,
Borneo and New Guinea using chloroplast DNA markers rps 2 gene, trn L intron and trn Ltrn F intergenic spacer.
This study suggested that the migratory direction between Taiwan and the Philippines is possibly from north to south, while the migratory direction within section Malesianae and the center of origin of Euphrasia remain unanswered. In this study, in order to give a full picture of the evolution of Euphrasia , we reconstructed a phylogeny of Euphrasia by integrating our data with the recorded data at GenBank submitted by Gussarova and her colleagues. Our results indicate that the Northern
Hemisphere origin of Euphrasia is not well supported. In addition, we found that the Asian tropical and subtropical alpine areas are a contact zone rather than a transition zone. Two lineages, one from the Northern Hemisphere and the other from the Southern Hemisphere, may be recognized. The section Malesianae oringinates in the
Northern Hemisphere and the section Pauciflorae oringinates in the Southern Hemisphere. The results support Barker’s opinion proposed in 1982. A different line of evidence, reconstruction of a phylogenetic tree of
Euphrasia using samples of eight species from 30 localities in the connecting areas including section
Malesianae from Borneo, Luzon and Taiwan, and section
Pauciflorae from New Guinea based on markers of three chloroplast DNA, nine mitochondrial DNA, and nuclear
DNA ITS, also supports a contact zone theory.
P1020 – ePoster
1
Revision on Chinese Lophocoleaceae
Wu, Y 1 , Gao, C 2
Hangzhou Normal University, China; 2 Institute of
Applied Ecology, Chinese Academy of Sciences
Lophocoleaceae includes Chiloscyphus, Heteroscyphus and Leptoscyphus and Physotheca , developing female involucres without involvement of stem tissue. Chinese
Lophocoleaceae is revised. Accoding to Piippo (1990),
30 species of Geocalycaceae are recorded for China, including 10 in Heteroscyphus Schiffn. and 16 in
Chiloscyphu s Corda. Gao and Wu (2008) recorded 38
Geocalycaceae in China, incuding one in Leptoscyphus ,
14 in Heteroscyphus Schiffn. and 16 in Chiloscyphus
Corda . 65 taxa (including 56 species, seven varieties, and two forms) of Lophocoleaceae have been reported from
657

China. Among them, 12 species, one varieties and one form of Lophocoleaceae were originally described as new based on specimens collected from China. Based on this visional study, three genera, 22 species, are considered to be taxonomically recognizable in China.
Seven new synomyns are made.
P1021 – ePoster
Evolvement of Hepaticae based on perichaetium morphology
Gao, C 1 , Wu, Y 2 , Li, W 1
1 Institute of Applied Ecology, Chinese Academy of
Sciences; 2 Hangzhou Normal University, China
Sexual reproductive process of Hepaticae is based on sexual organs of male antheridiophore and female archegonium. Perigonia dies after fecundation with ovum. In contrast, after fecundation of ovum, in order to protect young sporophytes till mature perichaetium that produces archegonium undergoes a series of developments and changes in shapes and compositions.
The differences of such development and changes among groups indicate the systematic development and evolvement processes of Hepaticae. Compositions of nearly mature Perichaetium include Sporophyte (Sp),
Calyptra (Ca), Bract (B), Bracteole (Br), Perianth (P),
Pseudoperianth (Ps), Perigynium (Pe), Coelocaule (Co),
Scale (S), Receptable (Re), Carpocephalum (Car) and
Involucre of Carpocephalum (Carin). Based on the compositions of Perichaetium, Hepaticae can be categorized into following types, perichaetium of calyptra type, perichaetium of perianth type, perichaetium of coelocaule-perianth type, perichaetium of coelocaule type, perichaetium of perigynium type, perichaetium of involucre type, and perichaetium of carpocephalum involucre type. The most original perichaetium is a large fleshy calyptra directly developed from the wall of archegonium after fecundation of ovum and during young sporophytes developed. Perichaetium of calyptra is consist of sporophyta and calyptra developed from archegonium. It further develops into perichaetium of perianth type. On the basis of perichaetium of calyptra, calyptra weakens forming a membrane of 1–2(3) cells covering young sporophytes.
Between calyptra and scales leaf trace forms perianth of bag- or barrel-shapes protecting young sporophytes till mature. It further evolves as perichaetium of coelocaule and perianth type. Perianth also weaked as well as calyptra. The tip of reproduction stems or reproduction branches produces hollow coelocaule of half length to which the young sporophytes sink in and develop and mature. Such a structure is evolvement to adapt to environment and better suitable to bad environment. On the basis of perichaetium of coelocaule and perianth type, it further develops as perichaetium of coelocaule type.
Perianth weakens and reproduction stems or reproduction branches further differentiates as hollow coelocaule.
Perichaetial leaves extend to the air. Based on perichaetium modality structure and evolvement trace of gametophyte of Hepaticae, Hepaticae can be categorized into five subclasses: Calyptrocoleidae, Jungermanniidae,
Perrssonilliaidae, Pallaviciniidae, and Marchantiidae.
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P1022 – ePoster
Segregation of the polyphyletic genus
Polyalthia
(Annonaceae)
Xue, B 1 , Su, Y 1 , Mols, J 2 , Kessler, P 3 , Saunders, R 1
1 University of Hong Kong, China;
The Netherlands;
Netherlands
3
2 Leiden University,
Hortus Botanicus Leiden, The
The circumscription of the species-rich genus Polyalthia
(Annonaceae, with ca. 150 species) has long been recognised to be highly problematic. The genus has functioned as a ‘dustbin group’ for species of unclear affinities due to the absence of clear synapomorphies, resulting in a morphologically heterogeneous species assemblage. Recent molecular phylogenetic analyses have indicated that Polyalthia is highly polyphyletic, with species scattered in multiple clades. The ‘ Polyalthia hypoleuca complex,’ including Asian Polyalthia species with columellar-sulcate pollen, was recently segregated to form a new genus, Maasia . A considerably broader taxonomic sampling of Polyalthia species and associated taxa is included in the present phylogenetic analyses, with the objectives of increasing tree resolution and statistical support, hence enabling the recognition of strictly monophyletic genera with clearly defined diagnostic morphological characters. Bayesian and maximum parsimony analyses of chloroplast mat K, rbc L and trn L-F DNA sequences consistently gave congruent topologies, with Polyalthia species distributed in five well-supported clades. Based on molecular and morphological data, existing Polyalthia species can be classified into the following five genera: (1) Fenerivia :
10 Madagascan Polyalthia species with columellarsulcate pollen form a well-supported clade distinct from
Maasia . The generic name Fenerivia is reinstated for this clade since the type species Fenerivia heteropetala (=
Polyalthia heteropetala ) is included. (2)
Marsypopetalum : five Asian Polyalthia species with thick inwardly-curved inner petals form a well-supported clade with Marsypopetalum pallidum (sister to
Trivalvaria ) and will be accordingly transferred to
Marsypopetalum . (3) Enicosanthum : several Polyalthia species which belong to Polyalthia section Monoon (with a solitary, basal ovule in each ovary) form a wellsupported clade (sister to Neo-uvaria ) together with some
Enicosanthum species, Woodiellantha sympetala and
Cleistopetalum borneense (a synonym of Polyalthia sinclairiana ). (4) A new, currently unnamed genus: based on a well-supported clade consisting of several
Polyalthia species, sister to Miliusa . (5) Polyalthia sensu stricto: the ‘true’ Polyalthia clade, including the type species, Polyalthia subcordata , and other species in
Polyalthia section Polyalthia (with more than two ovules per ovary) as well as species in the small Australian endemic genus Haplostichanthus . The name
Haplostichanthus will accordingly be recognized as a synonym of Polyalthia sensu stricto.
P1023 – ePoster
Biogeography and biosystematics of family
Hyacinthaceae in India
Yadav, S 1

1 Dept of Botany, Shivaji University, Maharashtra, India
Family Hyacinthaceae mainly distributed in Africa is represented by 4 genera and c. 20 species in India.
Extensive literature and field survey, cytological studies and experiments on hybridization have been conducted to understand biogeography and biosystematics of the family Hyacinthaceae in India. Ledebouria hyacinthiana was wrongly identified and described under the genus
Scilla . Moreover, Scilla viridis is conspecific to
Ledebouria hyacinthiana . Ledebouria hyacinthiana widely distributed especially in peninsular India shows great polymorphism and has three cytotypes viz. 2n = 30,
45, and 60. The triploid (2n = 3x = 45) population is adapted especially to plateaus of higher altitudes in northern Western Ghats. It shows vegetative propagation through formation of bulbils at leaf tips . Three cytotypes found in northern Western Ghats show particular distribution pattern i.e. the diploids in plains, triploids at higher altitude and hexaploids in extremely dry regions.
Scilla hohenackeri is reported from western Himalayan region. Genus Dipcadi is represented by c. 9 species
(there are some disputes about identity of some species) of which 7 are found in northern Western Ghats and 5 are endemic to the region. The flower odor from foul to sweet in various Indian species appears to be major cause in differentiation of populations leading to speciation.
Cytological studies in 7 species revealed that D. concanense, D. goaense, D. saxorum and D. serotinum possess 2n = 12; D.
montanum and D. ursulae have 2n =
20 and D . erythraeum has 2n = 22. Crossing experiments showed close genetic relationship between D. montanum and D. ursulae. Genus Drimia is represented by c. 9 species, however, identity of some species is doubtful .
Hooker reported D. polyphylla , which seems to be anomalous form of D. coromandeliana which very rarely possesses leaves and flowers together in few individuals.
Based on flower blooming, the species can be divided into two groups i.e. day blooming ( D. polyantha , D. razii and D. wightiana ) and night blooming ( D. indica and D. coromandeliana ). Among the species, D. indica shows three cytotypes viz. 2n = 20, 30 and 40. The triploids are restricted to saline sea shores. Drimia wightiana shows two cytotypes with 2n = 20, and 40. Drimia coromandeliana has 2n = 40 and reproduces sexually but with some sterility. Drimia polyantha and D. razii both are diploids with 2n = 20. Drimia polyantha populations of higher altitudes show up to 7 accessory chromosomes.
Crossing experiments showed close relationship among all the Indian species of the genus except for D. wightiana which failed to cross with any of the Indian species in vice-versa crosses. High endemism and the highest concentration of species of Hyancinthaceae and their cytotypes conclusively suggest that the northern
Western Ghats are the centre of diversification for the family in India. This communication presents the observations on distribution, species identity, cytology and breeding behavior of members of family
Hyacinthaceae in India to understand their biogeography and biosystematics.
P1024 – ePoster
Digitization of forest pathology herbarium: a bioinformatics tools development in forest pathology
Yadav, N 1 , Samanta, S 1
1 Forest Research Institute, India
The present era has seen an exponential increase in the growth and diversification of all forms of information, which is sometimes called as information explosion. This has been made possible due to the impact of information
& communication technology on the modern society.
One of the major benefits of this revolution has been the evolution of Biodiversity Informatics. Plant and animal specimen data held in museums and herbaria, survey data and species observational data provide a vast information resource, providing not only present day information on the locations of these entities, but also historic information going back several hundred years (Chapman and Busby 1994). It is estimated that there are approximately 2.5–3 billion collections worldwide in museums, herbaria and other collection institutions
(Duckworth et al. 1993, OECD 1999). A key purpose of digital information in the biological sciences is to provide users of information with a cost-effective method of querying and analysing that information. The biological world is infinitely complex and must be generalised, approximated and abstracted in order to be represented and understood (Goodchild et al. 1991).
Ways of presenting biodiversity information to users is through the use of geographic information systems, environmental modelling tools, decision support systems, books, CDs, images and on-line databases, specimens and their parts, DNA reports, etc. Forest Mycology got its initiation as a section of Botany Branch in 1927. Later this section was elevated to full fledged Forest Pathology
Branch in 1950. Since the inception of this discipline, pioneering studies were carried out on many disease problems relating to plantations, nurseries and timber storage yards. The Herbarium was enriched by the collections of Dr. K.D. Bagchee and B.K. Bakshi from the country . Apart from this it received specimens from
Pusa Herbarium, Mycological Herb. Ottawa, Canada,
Beltsville, Washington, USA Flora Exsiccatta Austro
Hungarica, U.K. and Bureau of Plant Industries USDA,
U.S.A. The project of pathology herbarium digitization aims at developing the database software system that can also be fruitfully utilized for research & teaching purposes. Generally the published records of fungal pathogens from most countries are unsupported by specimens or cultures from which the identity of the pathogen could be checked. Many common pathogens are poorly represented in large herbaria and culture collections. The herbarium thus becomes an important tool in revision of taxa or making monographic studies of a particular genus. The Herbarium at Forest Pathology
Division housing nearly twelve thousand specimens and it is the largest in South East Asia. It has collections of forest fungi from all the agro climatic zones of the country. The digitization work is aimed for developing the state of art software database as per international biodiversity informatics guidelines and standards.
P1026 – ePoster
Evolutionary process of reproductive isolation in bird-nest fern
Asplenium nidus
and its related species
Yamada, K 1 , Kakugawa, Y 2 , Kato, H 1 , Murakami, N 1
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1 Makino Herbarium, Tokyo Metropolitan University,
Japan; 2 Botanical Gardens, University of Tokyo, Japan
Species is the most basic and real unit in taxonomy. At present, most commonly accepted definition of species is the biological species concept that defines species as
‘groups of actually or potentially interbreeding natural populations, which are reproductively isolated from other such groups’ (Mayr 1942). Following biological species concept, reproductive isolation is the most important for defining species. Asplenium nidus L. and its relatives ( A. nidus complex) are a group of epiphytic fern species with simple leaves. It was reported that many cryptic species
(morphologically hard to distinguished but reproductively isolated to each other) are contained in the complex. It was also elucidated that the genetic distances based on rbc L sequences and the degree of reproductive isolation among the cryptic species are positively correlated (Yatabe et al. 200#). Therefore, I considered that Asplenium nidus L. complex might be one of the best materials to reveal the evolutionary process of reproductive isolation, which is the main process of speciation. During the period of master course, I have two kind of research as follows up to the present: searching members of A. nidus complex which are on the process of speciation in Japan and Developing codominant nuclear DNA markers, which will enable me to estimate parents of zygotes or small embryos. I collected plant samples of Japanese species of the complex: Asplenium antiquum, A. nidus and A. setoi from 15 points in various localities of Japan, also observing their natural habitats. Then, I determined their rbc L sequences. As the result, even between the samples of A. setoi collected in Ogasawara Islands and those in
Ryukyu Islands, up to 6 base pair difference was observed. As Yatabe et al. (2009) reported that reproductive isolation was observed among individual of the complex whose rbc L sequences differed in 5–6 bases, we can expect that at least partial reproductive isolation might exist among individuals of A. setoi in Ogasawara
Islands and those in Ryukyu Islands. I also tried to develop SSR, gapCp and pgiC markers during period of my master course. Then, I succeeded in developing PCR primers to amplify pgiC marker. Obtained nucleotide sequences of pgiC gene using the developed primers, showed enough amount of variation even among the samples of Asplenium setoi from various localities in
Japan. In near future, I am planning to conduct artificial crossing experiment between Asplenium setoi from
Ogasawara Islands and those from Ryukyu Islands.
Moreover, I will observe detailed process from fertilization to embryogenesis during artificial crossing among various combination of A. nidus complex in Japan and from adjacent foreign countries, using digital microscope and other facilities.
P1028 – ePoster
Paleovegetation of fossil wetland forests dominated by
Metasequoia
and
Glyptostrobus
from the lower
Pleistocene Kobiwako Group, Japan
Yamakawa, C 1 , Momohara, A 2 , Matsumoto, M 3
Nunotani, T 1
,
1 Lake Biwa Museum, Japan; 2 Graduate School of
Horticulture, Chiba University, Japan; 3 Graduate School of Science, Chiba University, Japan
An Early Pleistocene (1.8–.9 Ma) fossil wetland forest community that was dominated by Metasequoia and
Glyptostrobus was reconstructed based on the species composition of in situ stumps and other plant macrofossil assemblages. The plant fossils were recovered from a fossil forest preserved in deposits of the Kobiwako
Group that are exposed in the Echi River, Shiga
Prefecture, central Japan. Fossil woods of Metasequoia and Glyptostrobus were distinguished based on anatomical characteristics that are the most distinctive differences of living trees; the presence of gaps between the ray cells and the arrangement and maximum height of the ray cell stacks. Distribution of the wood fossils from different horizons in the fossil forest indicates
Metasequoia grew over a long period of time in a stable environment, while Glyptostrobus and Alnus grew in unstable environments characterized by short-interval floods. The fossil forest as a whole represents a fluvial back-marsh environment, based on sedimentary facies.
Aquatic and wetland herbs, such as Cyperus , Carex ,
Scirpus, Polygonum, and Menyanthes , grew in and around the wetland forest. The upland forest was composed of mixed evergreen conifers and deciduous broad-leaved trees, including Chamaecyparis pisifera,
Tsuga, Magnolia, Cornus , and Acer . The assemblage of fossil plants recovered from the fossil forest consists of plants that are currently distributed in the cool temperate climate zone, such as Picea , Thuja, Betula maximowicziana, Pterocarya rhoifolia, and Menyanthes trifoliata . Glyptostrobus is distributed only in subtropical areas at present, but it also grew under a cool temperate climate in Japan during the Early Pleistocene.
P1029 – ePoster
Do apogamous fern
Dryopteris erythrosora
obtain genetic variation from its related sexual species?
Yamamoto, K
Murakami, N 1
1 , Yatabe-Kakugawa, Y 2 , Ebihara, A 3 ,
1 Makino Herbarium, Tokyo Metropolitan University,
Japan; 2 Botanical Gardens, Graduate School of Science,
University of Tokyo, Japan; 3 Dept of Botany, National
Museum of Nature and Science, Japan
In apogamous reproduction of ferns, unreduced spores are formed and resultant gametophytes from the spores produce sporophytes of next generation without fertilization. Therefore, all offspring sporophytes from an apogamous parent are expected to be genetically identical. However, apogamous ferns frequently show large morphological and genetic variation. Walker (1962) hypothesized that it is because apogamous species obtain genetic variation of related sexual species through hybridization. Most apogamous fern species are triploid
(3x). However, when 3x apogamous species hybridize with diploid (2x) sexual species, tetraploid (4x) will be produced. Lin et al (1995) hypothesized that there occur recurrent ploidy reduction from 3x to 2x apogamous ferns, and increase their ploidy from 2x to 3x by the crossing with related sexual 2x species. If such the cycle occurs, 3x apogamous fern species can obtain genetic
660

variation from its related 2x sexual species without polyploidization. In order to test the cycle hypothesis, we should reproduce all steps of the cycle hypothesis under experimental conditions. It is also necessary to examine natural mixed population of closely related sexual and apogamous fern species. We selected 3x apogamous
Dryopteris erythrosora and its closely related 2x sexual
D. caudipinna as materials. In this study, we tried to reproduce following 2 steps of the cycle hypothesis on culture media and also examined their possible frequencies: (1) hybridization between the 3x apogamous and the 2x sexual species and (2) ploidy reduction from the 3x apogamous type to 2x apogamous type through unequal meiosis. For the purpose, we conducted following sets of artificial crossing experiments including controls: (a) 3x apogamous type (

‰)

~ 2x sexual type (

Š); (b) 2x sexual type (

‰)

~ 3x apogamous type (

Š); (c) 3x apogamous type (

‰)

~
3x apogamous type (

Š); (d) 2x sexual type (

‰)

~
2x sexual type (

Š); (e) isolated gametophyte of apogamous type; (f) isolated gametophyte of sexual type;
(g) sowing only apogamous type spores; (h) sowing only sexual type spores. In order to detect formed hybrids or ploidy-reduced apogamous offspring, we analyzed both ploidy and genotype of the formed offspring. For ploidy estimation, we used ploidy analyzer. Genotypes of offspring were examined using allozyme markers (PGI enzyme). In addition, we both cytologically and genetically examined the natural mixed populations in
Izu-Ohshima Island. We at first estimated reproductive mode by counting spore numbers per sporangium (64 spores, sexual; 32 spores, apogamous). Next, genotype of each plant was examined using PGI allozyme analysis. In
Izu-Ohshima, 8 mixed population of the apogamous and sexual types and 22 pure apogamous populations were found. Amount of genetic variation observed in apogamous plants of the mixed populations were significantly higher than those of the pure apogamous populations. Thus, it was supported that the apogamous fern obtained genetic variation of the related sexual species in nature.
P1030 – ePoster
Molecular phylogeny of the Himalayan genus
Erioscirpus
(Cyperaceae)
Yano, O 1 , Ikeda, H 1
1 Dept of Botany, University Museum, University of
Tokyo, Japan
The genus Erioscirpus Palla (Cyperaceae) is a
Himalayan genus and comprises two species, E. comosus
(Wall.) Palla and E. microstachyus (Boeck.) Palla. This genus has often been treated as a synonym of
Eriophorum or Trichophorum by having cotton hair-like long perianth bristles of achenes. However, Erioscirpus was quite different from Eriophorum and Trichophorum species in their morphology and ecology. In this study, we aim to clarify the phylogenetic position of the genus
Erioscirpus . We asked specifically: (1) Is Erioscirpus species included in the genus Eriophorum or
Trichophorum ? (2) If not, what is closely related genus to Erioscirpus ? 3) Where is the origin of Erioscirpus ?
We analyzed molecular phylogeny using chloroplast
DNA trn L intron, trn L-F spacers, and nuclear DNA ITS sequence data of E. comosus from Nepal and Myanmar, and E. microstachyus from India. Our data have revealed that Erioscirpus is most closely related to members of tribe Cypereae. In the tribe Cypereae, Erioscirpus was placed as related group consisting the genera Hellmuthia,
Isolepis, Ficinia, Desmoschoenus , and Scirpoides which is mainly distributed in Southern Hemisphere. On the other hand, the genera Eriophorum and Trichophorum formed a monophyletic clade with members of tribes
Cariceae, Dulichieae, or Scirpeae which is mainly distributed in Northern Hemisphere. Therefore, our results suggested that Erioscirpus and
Eriophorum / Trichophorum might have been arised from different origin. Erioscirpus might be originated in tropical or warm-temperate regions of Southern
Hemisphere.
P1032 – ePoster
Anatomo-morphological and molecular studies of
1
Ergocarpon and
Anisosciadium
(Umbelliferae-
Apioideae-Echinophoreae)
Zakharova, E 1 , Kljuykov, E 1 , Degtjareva, G 1
Moscow State University, Biological Faculty, Botanical
Garden, Russia
The genera Ergocarpon and Anisosciadium belong to the tribe Echinophoreae, one of relatively few distinct and apparently homogeneous tribes of the family
Umbelliferae. Tribe Echinophoreae is distributed mainly in Mediterranean, Southwest Asia and Middle East and includes 6 genera: Anisosciadium, Dicyclophora,
Echinophora , Ergocarpon , Pycnocycla and Thecocarpus .
The tribe is characterized by a very distinctive type of umbellule whith sexual differentiation of flowers, varying degrees of fusion and induration bracteoles, pedicels and fruits. The structure of the inflorescence in many members of the Echinophoreae is studied well enough (Hedge & Lamond 1973). However, anatomical fruit structure remains very poorly known. The present study aims to examine morphological and anatomical characters of fruits in genera Ergocarpon and
Anisosciadium . Central fruit (immature) of Ergocarpon cryptanthum (Rech.f.) C.C.Townsend half sunk in receptacle and up to the middle fused with sterile flowers; fruit apex free, pubescent; calyx teeth absent; stylopodia low-conical; styles spreading. In transverse section, mericarps slightly compressed laterally or nearly rounded, commissure broad; mesocarp parenchymatous; vittae solitary (rarely 2–3 in valleculae, 2–4 on the commissure); rib secretory ducts solitary, large; vascular bundles broad; seed face double-grooved. Central fruit
(mature) of Anisosciadium orientale DC. sessile, free on the receptacle, oblong-ovate, pubescent; calyx teeth absent; stylopodia low-conical; styles spreading. In transverse section, mericarps nearly rounded, commissure broad; hypoderm from 2–3 layers of collenchyma cells; mesocarp parenchymatous; vittae solitary in valleculae, 2 on the commissure; rib secretory ducts absent; vascular bundles broad; endocarp from 2–3 layers of extended in tangential direction cells; seed face with deep groove. Thus, genera Ergocarpon and
Anisosciadium have similar fruit anatomical structure.
However, they clearly differ in an inflorescence structure. Ergocarpon has large, triangular bracteoles,
661

much longer than the umbellules and hiding the flowers.
Central flower is hermaphrodite, outer flowers are sterile, occasionally hermaphrodite. Fruit is half sunk in receptacle and up to the middle fused with sterile flowers. Unlike Ergocarpon , Anisosciadium has foliaceus or spinescent bracteoles, more shortly the umbellules, free or fused with outer flowers. Fruits are sessile, free on the receptacle. Central fruit is bicarpellate, outer fruits are monocarpellate only. Some flowers are sterile. In addition, the nuclear ribosomal ITS
DNA data for Ergocarpon cryptanthum was obtained and compared with other genera of the tribe Echinophoreae, representing a monophyletic group according to ITSbased phylogeny of Umbelliferae (Downie et al. 2010).
Molecular phylogenetic analysis confirmed that
Ergocarpon is a part of this tribe suggested previously by morphological data. It occupies isolate position within the Echinophoreae clade and more close to Dicyclophora and Pycnocycla , whereas Anisosciadium is allied to
Echinophora .
P1033 – ePoster
Pollen morphology of the genus
Orobanche
(Orobanchaceae) from Turkey
Zare, G 1 , Oybak Dönmez, E 1 , A. Dönmez, A 1
1 Hacettepe University, Dept of Biology, Ankara, Turkey
Orobanche is the largest genus among the holoparasitic members of Orobanchaceae that is mainly distributed throughout the subtropical and temperate regions of the northern hemisphere, and the Mediterranean region is one of the most important centers of diversity. In this study pollen morphology of Orobanche species found in
Turkey have been examined. A comparative palynological study of 128 specimens of the genus
Orobanche , representing all species, and several species of other genera of the family, such a Pedicularis,
Cistanche and Phlypaea , have been studied and illustrated with light (LM) and scanning electron microscopy (SEM). The grains are typically isopolar, radially symmetrical, more or less spheroidal and there are two basic types of pollen grain in Orobanche genus, tricolpate and inaperturate, but in some species there are
2-syncolpate and 3-syncolpate pollen grains. The
Orobanchaceae pollen grains display considerable variation among genera and species, sometimes with taxonomically significant characters at the levels of genus and species. A clear difference between the pollen grains of the section Trionychon and the section
Orobanche has been noticed. The results obtained highlight the division of the genus into two sections
( Trionychon and Orobanche ), as has been traditionally recognized (Back von Mannagetta 1930) . The results of the SEM studies show the major pollen types, based on exine ornamentation. In these major pollen groups there are also minor types. A key to pollen morphology of the studied taxa within Orobanchaceae has been made based on pollen morphology, and the taxonomic relationship of this family has been discussed.
662
P1034 – ePoster
Genetic diversity and phylogeography of Chinese endemic genus
Dipteronia
Oliv. (Aceraceae) inferred from chloroplast (cp) DNA
Zhao, G-F 1
1 College of Life Sciences, Northwest University
The genus Dipteronia Oliv. is endemic to China, and consists only two species, D.sinensis
Oliv. and D. dyerana Henry. In this study, two chloroplast (cp) DNA intergenic spacers, rpl 20rps 18 and trn V intron, were employed to analyze genetics diversity and phylogeographic pattern of Dipteronia Oliv. Based on sequence variation of the 90 individuals from 18 populations, 19 haplotypes were detected in total.
Relatively high level of haplotype diversity (h=0.716) and nucleotide diversity (Pi=0.007) were detected in
Dipteronia Oliv. AMOVA analysis revealed that the majority of the genetic variation was partitioned among populations within D.sinensis
Oliv. and D. dyerana
Henry. The molecular phylogenetic data, together with the geographic distribution of the haplotypes, suggested the existence of a strong phylogeographic pattern (NST =
0.802, GST = 0.499, NST > GST; U = 0.25). Based on relationships among haplotypes (most parsimonious tree and the 95% plausible network), together with mismatch distribution analysis and neutrality tests, we conclude that restricted gene flow with isolation by distance and allopatric fragmentation should be responsible for the present distribution pattern of Dipteronia Oliv.
(Aceraceae). However, we do not find evidence of population expansion in both D.sinensis
Oliv. and D. dyerana Henry. What’s more, multiple localized glacial refugia around Sichuan Basin (in Mts. Qinling, Mts.
Bashan and the south edge of Yunnan-Guizhou Plateau) were found.
P1035 – ePoster
Occurrence of anisospory in hornworts, with special reference to the status of
Hattorioceros
(Anthocerotophyta)
1
Zhu, R-L 1 , Peng, T 1 , He, Q 1
East China Normal University, China
Hattorioceros (J.Haseg.) J.Haseg., a monospecific genus, was established based on H. striatisporus J.Haseg. from
Himachal Pradesh (Western Himalaya), India. As originally described and illustrated, the unique characteristic for recognition of Hattorioceros is its unusual spores with canaliculated-striate surfaces, which do not occur in other hornworts. Our investigation found that two kinds of spores (large, tetrahedral spores with papillate surfaces, and small, non-tetrahedral spores with canaliculated-striate surfaces) sometimes occur in the same capsule of Hattorioceros striatisporus . The anisospory was found in hornworts for the first time.
Although spore morphology is considered to be extremely important in systematics of hornworts, it must be careful to establish higher taxonomic ranks only based on a sole spore morphology character. Owing to absence of essential features except for spore morphology,

Hattorioceros is here proposed as a new synonym of
Phaeoceros , and its type species, H. striatisporus , is reduced to the cosmopolitan P. carolinianus (Michx.)
Prosk.
P1037 – ePoster
Functional diversity of woody plants in West Africa
Schmidt, M 1, 4, 5
Mbayngone, E
, Traoré, S
3 , Ouédraogo, O 2 , Hahn, K
1
A 2 , Zizka, G 1, 4, 5
2 , Ouédraogo, A
1
2 , Zizka, A 1 ,
, Thiombiano,
2
Senckenberg Research Institute, Frankfurt, Germany;
University of Ouagadougou, Burkina Faso; of N'Djamena, Chad; 4
3 University
Goethe University, Frankfurt,
Germany; 5 Biodiversity and Climate Research Centre,
Frankfurt, Germany
West Africa is climatically characterized by a steep precipitation gradient that is reflected in the vegetation zones from the Sahara Desert to the Lower Guinean rainforests. The abundance and diversity of woody plants increases from the north to the south. While the savannaforest boundary is usually characterized by abrupt changes and mosaics in the areas of transition, the transition between different savanna types in the
Sudanian and Sahelian zones is continuous. In our study we focus on Burkina Faso, a land-bound country in the center of West Africa that can in many ways be seen as a model case for the whole region. Burkina Faso is rather flat with an altitudinal range of 600m between the lowest areas in the floodplains of the Pendjari River and the highest peak at Mt Ténakourou. Its vegetation is characterized by Sahelian and Sudanian savannas with acacias dominating the woody flora in the north and
Combretaceae replacing them continually towards the south. Some Guinean elements are present in the gallery forests and the few remaining forests islands in the southwest. Woody plants in Burkina Faso are still the main source of energy, medicine and other NTFPs and provide various ESS including soil preservation and water retention, so people are highly dependent on them.
As primary producers and structural habitat components they are the base of the region’s ecosystems. Since woody plants are important both ecologically and economically, the aim of this study is to investigate in detail the geographical patterns of the most frequent woody species of Burkina Faso. Botanical distribution data is very unevenly distributed throughout our study area. Most data is confined to sampling hotspots, regional studies often had also a focus on particular habitats or species groups. Therefore we modeled species distributions of 129 woody plant species using remote sensing vegetation indices, climatic and topographic predictor variables with the program Maxent to extrapolate the distributional information to the whole area of Burkina Faso. In a later step we counted presences within each single grid cell for all species and for species groups defined by functional traits or higher taxa. For numerical traits we calculated the mean values for the single grid cells. The species richness of woody plants increased from the Sahelian to the Sudanian Zone with highest values in the Northern Sudanian Zone.
Apart from the zonal trends, local patterns can also be observed, e.g. higher species richness in areas of high topodiversity. Clear patterns can be shown for several traits: Spinescence is decreasing from the north to the south. Species containing latex are mainly confined to the Sudanian Zone with highest values in sandstone mountains. Compound leaves are characteristic for about half of the species throughout the country. Only in the south-west a lower share of species with compound leaves is observed. Flowering phenology has strong latitudinal trends with most northern species flowering in the rainy season and most southern species in the dry season.
P1038 – ePoster
The
Gravisia
/
Portea complex (Bromelioideae,
Bromeliaceae) – new insights from molecular studies
Heller, S 1,4 , Leme, E 2 , Schulte, K 3 , Zizka, G 1,4
1 Senckenberg Research Institute and Goethe University,
Frankfurt, Germany; 2 Herbarium Bradeanum, Rio de
Janeiro, Brazil; 3 Australian Tropical Herbarium, James
Cook University, Cairns, Australia; 4 Biodiversity and
Climate Research Institute, Frankfurt, Germany
Phylogenetic relationships among the core
Bromelioideae (Bromeliaceae) are still not resolved sufficiently. Especially the group of Aechmea and allied genera poses still a number of systematic/taxonomic, nomenclatural and phylogenetic problems. By applying highly variable AFLPs to assess genetic diversity, we tried to resolve relationships in morphologically characterised groups, which have been, at least in part and at various times, grouped in separate genera . We investigated 73 species from the genera Aechmea,
Ananas, Canistrum, Canistropsis, Edmundoa,
Eduandrea, Hohenbergia, Neoregelia, Nidularium,
Portea and Quesnelia . The analysis of 11 primer pairs revealed over 2500 characters that were scored manually and automatically (using the program GENEMARKER
V1.90). Different analyses (Neighbor-Joining, Maximum
Parsimony, Maximum Likelihood, Mr Bayes) were performed and the results combined in a synthetic tree
(majority rule consensus tree). Selected characters of the species (habitat, petal colour, biogeography, pollen morphology) were investigated by EL, partly extracted from literature and plotted on the tree . The species identified by Read & Luther 1991, Leme & Filho 2007,
Leme 2010 (pers. comm.) as the Gravisia/Portea complex and characterized by polyporate pollen were identified – for the first time based on molecular data and a fairly comprehensive sampling – as a monophyletic clade, nicely supporting the results of these authors.
Polyporate pollen appears to be a synapomorphy of the group, whose species have been placed in various genera like Aechmea, Canistrum subgen. Canistrum and Portea .
Within the complex two lineages could be identified, differing in their flower color and distribution. A further clade we detected comprised species of Canistrum subgen. Cucullathantus and the Aechmea fosteriana complex (Leme & Filho 2007) . Accessions of Canistrum subgen. Canistrum grouped not within the
Gravisia / Portea complex but were found to be genetically more similar to groups of Aechmea and the genus Hohenbergia.
663

P1039 – ePoster
Quiinaceae – a taxonomic online database
1
Schneider, JV 1,2 , Schmidt, M 1 , Herzog, E 2 , Zizka, G 1
Senckenberg Research Institute and Goethe University,
Frankfurt/Main, Germany; 2 Biodiversity and Climate
Research Institute, Frankfurt/Main, Germany
Monographs in print media are essential, but often have the disadvantages to restrict the citation of specimens to representative collections per geographical grid cell, to lack flexibility in respect of identification keys, and to be usually limited in depicting plants and specimens.
Particularly for species-rich genera with numerous rather nondescript or highly variable species a strictly dichotomous key is often not suitable. Here, we used the data accumulated during the taxonomic revision of the
Quiinaceae (Schneider & Zizka, submitted) – a family of
4 genera and 46 species distributed from Belize to South
Brazil and Bolivia – to develop an exemplary online data base that provides relevant taxon and specimen information complementary to the printed revision. Data retrieval is by browsing the taxa or by searching the requested data, e.g. collector and collection number of a specimen. Distribution maps are generated automatically for each taxon. Additionally, species identification is aided by an interactive multientry determination key and a considerable number of specimens is available as high resolution scan.
P1040 – Poster
Clarifying phylogenetic relationships and biogeographic events within Malagasy
Euphorbia
L. – a textbook example: the endemic subg.
Lacanthis
(Raf.) M.G.Gilbert, the island’s most diverse group
Aubriot, X 1 , Haevermans, T 1 , Ranaivo, J 2 , Lowry II, PP 3
1 Muséum National d'Histoire Naturelle, Origine
Structure et Evolution de la Biodiversité (OSEB), Paris,
France; 2 Centre National de Recherches sur l’Environnement, Madagascar;
Garden, St Louis, USA
3 Missouri Botanical
Euphorbia subgenus Lacanthis , as defined by
Haevermans in 2004 (from Gilbert’s 1987 circumscription), entirely endemic to Madagascar, comprises ca. 120 taxa (infraspecific taxa included), i.e.
>70% of the native members of the genus. Known for their spiny and succulent habit, these species show a tremendous array of morphologies (spiny or non-spiny bushes, monocaulous plants, geophytes…). One cultivar belonging to this group, the ‘Crown-of-Thorns’, is a well known horticultural plant and several geophytes are highly sought after by succulent collectors. The principal features of subg. Lacanthis are developed and brightly colored cyathophylls, prominent stipular structures
(crests, spines...) and succulence of the stems, leaves and roots. Members occupy a wide range of ecosystems, from rainforests to sub-arid and dry forests. The most recent monograph (Boissier, 1862) covered only a few species of this group, which were placed in different sections within Euphorbia based on the presence or absence of spines. Recent molecular analyses based on nuclear and
664 chloroplast markers (ITS, ndhF) suggest the paraphyly of such groupings, and support the monophyly of subg.
Lacanthis . However, these studies suffered from undersampling of taxa and characters, which resulted in poor node support within the group and its nearest relatives. In order to address biogeographic issues and understand radiation events, we have improved phylogenetic resolution by expanding taxon and character sampling, using two nuclear loci (ITS and ETS), and six chloroplast regions ( psb Atrn H, atp Iatp H, ndh A, mat K, rbc L, trn Qrps 16), with broad sampling from the subgenus. The results reveal incongruities between the markers and suggest that the chloroplast markers are too conserved to be of value for analyses at this taxonomic level. The ITS phylogeny is the only one consistent with results from previous studies, showing that the geophytic members of subg. Lacanthis (formerly recognized as sect.
Rhizanthium Boiss.) are polyphyletic. Phylogenies from the various markers still support the monophyly of subg.
Lacanthis , but the internal relationships within the group remain weakly supported or unresolved. The earliest diverging clades comprise species exclusively from northern and eastern Madagascar that share distinctive morphological and ecological traits within the subgenus: their stipules and cyathophylls are reduced, and they occupy restricted areas that present rather atypical environmental conditions for Euphorbia (karstic cliffs, rainforests…). Based on our phylogenetic results, we are now estimating the contribution of each molecular marker and conducting incongruity tests to compare the resulting topologies. Several rapidly evolving low-copy nuclear markers (Adh, Lfy, Sqd1, G3PDH) are being tested and amplified in an attempt to strengthen phylogenetic resolution within subg. Lacanthis .
P1041 – Poster
The contribution of DNA barcoding for the identification of Malagasy
Euphorbia
L. listed in
CITES Appendices I and II
Aubriot, X 1 , Haevermans, T 1 , Ranaivo, J 2 , Lowry II, PP 3
1 Muséum National d'Histoire Naturelle, Origine
Structure et Evolution de la Biodiversité (OSEB), Paris,
France; 2 Centre National de Recherches sur l’Environnement, Madagascar; 3 Missouri Botanical
Garden, St Louis, USA
Euphorbia L., the second largest and most diverse plant genus (with ca. 2000 species), is present in almost every part of the globe. The island of Madagascar, with at least
170 species of Euphorbia , almost all endemic, is a key hot-spot for the genus. All native species are listed in
CITES Appendix I and II, which controls international trade. All taxa on Appendix I belong to the endemic subgenus Lacanthis (about 120 taxa), which includes the life forms most critically threatened by trade (i.e., spiny and succulent species as well as geophytes). These plants are particularly difficult to identify due to the absence of a reliable revision or an authoritative reference work (the most recent treatment is that of Boissier in 1862).
Moreover, most taxa were described by amateurs in obscure publications, making identification even more problematical. DNA Barcoding methods may offer a promising tool for overcoming this problem, thus making it possible to monitor Euphorbia species under CITES.

Clarifying the chaotic taxonomy and nomenclature of
Euphorbia could also benefit from DNA barcodes.
Malagasy Euphorbia have not previously been included in Barcoding efforts. We tested this approach using at least two samples from each of 46 Malagasy Euphorbia species. Both of the core Barcoding markers for plants
(the chloroplast markers mat K and rbc L) were amplified and analyzed, and two additional candidate markers were also tested (chloroplast psb Atrn H and nuclear ITS2, both more variable that the officially recognized markers). The discriminative power at the species level for each marker and a combined dataset was estimated.
ITS2 is the only one to provide adequate discrimination among the taxa sampled: when performing a ‘Best match’ test, 80% of the identifications were correct. The two core barcodes ( mat K and rbc L) as well as psb Atrn H have poor discrimination unless used in combination with ITS2. The precise contribution of each marker taken alone or combined is estimated. Barcoding provides a new tool for the taxonomist when undertaking revisions and assessing species limits. Several well documented taxonomic problems within Malagasy Euphorbia were confirmed and clarified thanks to DNA Barcoding studies. A discussion is presented of the delimitation of species and infraspecific using both DNA Barcoding data and information from morphology and biogeography.
Results from this first-ever Barcoding test on Malagasy
Euphorbia show that this approach represents a useful tool for both identifying plant material and conducting taxonomic research.
P1042 – Poster
Evolutionary inference from multiple incongruent
DNA data matrices: reticulate evolution of
Polystachya
(Orchidaceae)
Russell, A 1 , Samuel, R 1
1
Chase, MW 2
, Barfuss, MHJ 1 , Turner, B 1 ,
Dept of Systematic & Evolutionary Botany, University of Vienna, Austria; 2 Royal Botanic Gardens, Kew, UK
The orchid genus Polystachya comprises approximately
250 species with an unusually wide distribution in the tropics and subtropics of Africa, Asia and the Americas.
Plastid DNA sequence analysis has provided good phylogenetic resolution and revealed contrasting patterns of endemism and dispersal in different lineages. Using data from a combination of nuclear and plastid loci, the current study significantly improves our understanding of
Polystachya phylogenetics and indicates the importance of reticulate evolution. This process, in which lineages have an interspecific hybrid origin, can be difficult to incorporate into evolutionary studies. We analysed ITS and plastid DNA sequences alongside three low-copy nuclear genes, and inferred hybrid origins of clades using i) incongruence between trees and ii) analysis of duplicated genes in tetraploids. Incongruence between gene trees was represented using supernetwork and consensus network software. Filtering was applied to reduce the influence of phenomena that might cause single gene trees to display clades not present in the others, such as deep coalescence, recombination or character homoplasy within individual datasets.
Relationships of tetraploids were inferred from cloning and sequencing homoeologous copies of low-copy genes, a procedure complicated by the tendency of similar but non-orthologous gene sequences to give rise to in vitro recombinants during PCR. Among the diploid accessions each data matrix gave a unique phylogenetic hypothesis, but filtered consensus networks had good resolution except among the basal branches, with little evidence of reticulation among diploids. Analysis of duplicated gene copies in tetraploids showed clearer evidence of hybrid evolution. One group of tetraploids showed evidence of multiple long-distance dispersals to achieve a pantropical distribution. A second group showed no evidence of multiple origins or long-distance dispersal but had greater morphological variation, consistent with hybridisation between more distantly related parents.
P1043 – ePoster
Functional implications of staminal lever mechanism in
Salvia cyclostegia
(Lamiaceae)
Zhang, B 1, 2 , Claßen-Bockhoff, R 3
, Li, Q-J 1
, Zhang, Z-Q 4 , Sun,
S 1 , Luo, Y-J 1
1 Key Laboratory of Tropical Forest Ecology,
Xishuangbanna Tropical Botanical Garden, Chinese
Academy of Sciences, Menglun Town, Mengla County,
China; 2 Graduate University of Chinese Academy of
Sciences, Beijing, China; 3 Institut für Spezielle Botanik,
Johannes Gutenberg Universität Mainz, Germany; 4 Key
Laboratory of Biodiversity and Biogeography, Kunming
Institute of Botany, CAS, China
Flower morphology and inflorescence architecture affect pollinator foraging behavior and thereby influence the process of pollination and the reproductive success of plants. Salvia is characterized by modified lever-like stamens and a dorsal pollination mode. This study explored possible ecological functions of the lever-like stamens and the floral design in Salvia cyclostegia .
Flower construction was experimentally manipulated by removing either the lower lever arms or the upper fertile thecae of the two stamens from a flower. The two types of manipulated individuals were intermixed with the control ones and randomly distributed in the population.
The Results indicated that removing the sterile lower lever arms significantly reduced handling time per flower of the main pollinator, Bombus personatus . Interestingly, this manipulation did not increase the number of flowers probed per plant visit, but instead reduced it, i.e. shortened the visit sequence of the bumble-bees. Both loss of staminal lever function by removing lower lever arms and exclusion of self pollen by removing upper fertile thecae significantly reduced seed set per flower and seed set per plant. Both the manipulations interacted significantly with inflorescence size for the effect on female reproductive output. It is concluded that the staminal lever, although reducing pollinator’s handling efficiency, to some extent, is of advantage as a mechanism for dispensing pollen, and that this particular floral design is associated with a remarkable floral constancy, with LLAs contributing to pollinator attractiveness and/or foraging cognition in S. cyclostegia .
P1044 – ePoster
Electrical signals in prayer plants (Marantaceae)
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Jerominek, M 1 , Claßen-Bockhoff, R 1
1 Institut für Spezielle Botanik, Johannes Gutenberg
Universität Mainz, Germany
The prayer plant family (Marantaceae) shows a unique pollination mechanism. In a split second the pollenpresenting style hits the pollinator and exchanges pollen.
This explosive style movement is based on a tension between the style and the hooded staminode. When a pollinator moves the hooded staminode the tension is released and the style rapidly springs forward. Thereby, water is transported through the tissue indicating a turgor change. The combination of mechanical tension and turgor movement raises the question whether the movement is released mechanically (as in Fabaceae) and/or electro-physiologically (as in Mimosa pudica and
Dionaea muscipula ). To test the two hypotheses we conducted apoplastic electrophysiological measurements and tried to release the style artificially by changing the membrane potential. For inducing a change of the apoplastic voltage we gave chemical (chloroform) and electrical stimuli to the upper surface of the style tissue.
In our test species Donax canniformis , we found a depolarisation of the apoplastic style membrane of up to
60mV. However, it was not possible to release the movement by only changing the membrane potential neither electrically nor chemically. Therefore we conclude: the style is mechanically released by pollinators and the observed depolarisation of the style membrane is the consequence of the corresponding loss of tissue tension (osmotic shock) and not the cause of the movement.
P1045 – ePoster
Evolutionary ecology of two African rain forest plant
1 genera:
Isolona
and
Monodora
(Annonaceae)
Couvreur, TLP 1 , Porter-Morgan, H 2 , Wieringa, JJ 3
Chatrou, LW 3
Institut de Recherche pour le Développement (IRD);
New York Botanical Garden, USA; 3
2
,
Nationaal
Harbarium Nederland, Wageningen Branch, The
Netherlands
The tropical rain forest (TRF) biome covers just ~7% of land but harbors over half of the planet’s biodiversity.
The TRF of Africa represents the second largest extent of the biome after the Amazon basin, and contains high levels of species diversity and especially endemicity.
Ecology is widely recognized as a major factor in speciation, thus a first step in unraveling African TRF diversification is to understand the role of ecology in explaining present day diversity. TRF diversification has been hypothesized to be the result of 1) geographical isolation followed by genetic drift and/or selection or 2) ecological gradients and divergent selection. In the former hypothesis we predict that ecological niches will be similar between sister species (niche conservatism), whereas in the latter ecological requirements will be significantly different. Here, we investigate ecological niche similarity between sister species within two TRF restricted plant genera of the pan tropical Annonaceae family, Isolona and Monodora , in order to quantify if ecology plays an important role in the diversification of
666
African rain forests in general. We generated a dated species level molecular phylogeny and identified a total of 11 potential sister species. Principal Component
Analysis of extracted bioclim variables and ecological niche modeling tests as implemented in EMNTools, based on over 1000 georeferenced specimens across
Africa and Madagascar, were used to test for ecological similarity. Our results indicate that most sister species had statistically similar ecological niches, even though they were never identical. In terms of diversification our results would imply that ecological speciation hasn’t played a major role in the evolution of Isolona and
Monodora species, even though some speciation events could be attributed to ecology. This contrast with several recent publications where significant ecological divergence was generally demonstrated for plant sister species in the such as in Lonicera , Cyclamen and five
Andean Solanum species. In addition, most speciation events within both genera were pre Pleistocene in origin.
This provides some support to the idea that present day diversity of African TRF originated before the
Pleistocene climatic fluctuations and that forest refuges have had more of an infraspecific genetic impact
(population level) and a limited influence on speciation.
P1046 – Poster
The Australian Plant Census: a model for consensus-
1 based taxonomic decision-making
Cowley, K 1 , Monro, A 1 , Lepschi, B 1
Centre for Australian National Biodiversity Research,
Canberra, Australia
The Australian botanical community is engaged in the construction of a ground-breaking census of its flora to be made available online. It involves all the major
Australian herbaria in decision-making at a national level, taking a consensus approach to resolving taxonomic and nomenclatural inconsistencies. For the first time in Australia, plant names from the level of kingdom to infraspecies are placed in an agreed classification to be updated in accordance with the latest scientific information . The Australian Plant Census
(APC) (http://www.anbg.gov.au/chah/apc/about-
APC.html) is a list of the accepted scientific names for native and naturalised Australian vascular plants. It includes all known synonyms and misapplications and distribution information for each taxon. A major strength of APC is its collaborative approach, with extensive input from taxonomic specialists throughout Australia and the endorsement of the Council of Heads of
Australasian Herbaria (CHAH). The consensus-based model encourages uptake of the APC concepts by contributing institutions and subsequent use at regional levels . APC builds on the foundation of the Australian
Plant Name Index (APNI)
(http://www.anbg.gov.au/apni/index.html), which is managed by the Australian National Herbarium (CANB) on behalf of Australia’s botanical community . APNI lists every published use of scientific plant names for the
Australian flora, including introduced species, but unlike
APC does not present information on recommended taxonomies or agreed nomenclature . The APC provides researchers, land managers, conservation agencies, regulatory agencies, industry and individuals with a

unified, authoritative, annotated inventory of the
Australian flora. When combined with the existing bibliographic data contained in APNI and linked to other databases such as Australia’s Virtual Herbarium, the
APC will be an unrivalled continental-scale resource for biodiversity information.
P1047 – Poster
Quantitative gene expression analysis in floral mutants of Buckwheat (
Fagopyrum esculentum
)
Demidenko, N 1 , Logacheva, M 1 , Fesenko, A 2 , Penin, A 1
1 Moscow State University, Moscow, Russia; 2 All-Russia
Research Institute of Legumes and Groat Crops, Orel,
Russia
Buckwheat, Fagopyrum esculentum , is a widely cultivated cross-pollinating pseudocereal and melliferous plant belonging to the family Polygonaceae (order
Caryophyllales). But despite of its importance in agriculture and food industry little is known about the genetic basis of development in buckwheat. Recently, the buckwheat floral transcriptome was sequenced and this facilitates molecular genetic studies in this plant, primarily the search of genes presumably involved in the control of flower development and the analysis of their expression. Quantitative reverse transcription PCR (qRT-
PCR) is one of the most precise and widely used methods of gene expression analysis. However, in order to obtain precise and reliable data accurate choice of reference genes is necessary. To find the optimal reference for gene expression analysis during buckwheat development we first studied the expression stability of eight candidate reference genes in different structures (leaves, inflorescences and fruits). Four genes – orthologs of
Arabidopsis At2g28390 (SAND family), At4g33380
(unknown function), At4g34270 (Tip41-like) and
At5g46630 (clathrin adaptor complex subunit) – were identified as stable. Using these genes as reference, expression profiles of several genes presumably involved in flower development were studied in three buckwheat mutants: green corolla – the mutant that is characterized by leaf-like tepals, fagopyrum apetala with carpelloid perianth and tepal-like bract, which has bracts similar to tepals. The orthologs of PISTILLATA (PI), AGAMOUS
(AG), APETALA1 (AP1), APETALA2 (AP2),
APETALA3 (the ABC-model genes), SEPALLATA,
LEAFY (LFY), WUSHEL (WUS), LEUNIG (LEU) and
SEUSS (SEU) were analyzed. FeAG expression was found to be increased in fagopyrum apetala and FeLFY – in tepal-like bract. B-class gene expression level was not changed in mutants with altered perianth (tepal-like bract, green corolla) suggesting that orthologs of B-class genes are not involved in the determination of petaloidy in buckwheat.
P1049 – Poster
A multigene phylogeny of pondweeds
(Potamogetonaceae)
Fehrer, J 1 , Kaplan, Z 1
1 Czech Academy of Sciences, Czech Republic
Aquatic plants exhibit large phenotypic plasticity and are characterized by a reduction of morphological characters.
These features are challenging for species determination and taxonomy. Poor species concepts and misidentifications in the Potamogetonaceae, one of the families of aquatic plants with the highest diversity, have additionally produced many wrong assignments and resulting relationships in some recent publications (and corresponding erroneous GenBank entries) that further complicate matters. In order to assist solving these problems and to establish species relationships in the framework of a monograph of the family at species level, we produced a multigene phylogeny based on two chloroplast ( rpl 20-5’ rps 12 and trn T-trnL intergenic spacers) and two nuclear ribosomal (ITS and 5S-NTR) markers consisting mainly of non-coding regions. One of the chloroplast and one of the nuclear markers each
( rpl 20-5’ rps 12 and ITS) were less variable while the other two ( trn Ttrn L and 5S-NTR) were highly variable and therefore most suitable to resolve close species relationships. Typically, multiple accessions per species were included, often from different continents. The family consists of three genera, Potamogeton (mostly 2x or 4x with x=13 or 14), monotypic Groenlandia (2x, x=15), and Stuckenia (6x, x=13) that are well distinguished by the molecular data. Outgroup species from other families were almost unalignable with most markers, especially their most variable regions. Also, the diversity among the three Potamogetonaceae genera is so large that even their alignments were ambiguous in many parts. Therefore, each genus was also subjected to phylogenetic analyses separately. Relationships of species groups were usually congruent among the different markers with the exception of two octoploid
Potamogeton species which differed in their placement between chloroplast and nuclear DNA trees. The relationships among different species groups somewhat depended on the marker used. The taxonomy and relationships of the linear-leaved species of Potamogeton
(2n=26 or 28) are particularly difficult because their morphology is even more reduced than that of broadleaved species (the majority of which is tetraploid).
While the less variable markers did not resolve them as a group, the more variable markers clearly showed them to be monophyletic. In most cases, intraspecific variation was very low with slight differences reflecting geographic origins. Some species were indistinguishable with all molecular markers although they can be distinguished morphologically. On the other hand, a few species comprised divergent (sometimes para- or polyphyletic) genotypes that usually corresponded to geographic origin although they do not show distinguishable morphological features. Hybrids could be clearly assigned to particular parental genotypes based on
ITS character additivity and were excluded from phylogenetic analyses. As we used the same markers as other researchers, we were able to correctly identify their wrongly assigned species by sequence comparisons. In the future we plan to add a low-copy nuclear marker
(adh1) to further elucidate relationships in this intricate group. When the taxonomic treatment of the species will be settled, we plan to establish an easy and reliable system for molecular species identification of the
Potamogetonaceae.
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P1050 – Poster
Nortriterpenoid glycosides from
Salicornia bigelovii
Torr. (Salicornioideae) and their chemotaxonomic significance
, Chen, Y 1 Feng, X 1 , Wang, M 1 , Wang, Q 1 , Shan, Y 1 ,
1
Liu, X 1
Institute of Botany, Jiangsu Province and Chinese
Academy of Sciences, Nanjng, PR China
Salicornia is a widespread, halophytic genus of family
Chenopodiaceae, subfamily Salicornioideae. The genus currently comprises 25 to 30 species occurring exclusively in the world. Some species of this genus are popularly known glasswort, saltwort, goosewort et al.
Salicornia is well known not only because of its unusual appearance with articulated succulent stems, but also because of its notoriously difficult taxonomy, which is considered rightly as a nightmare. Over the last 250 years, there is still no satisfactory taxonomic treatment, and it is frequently impossible to assign published information specifically to taxa within Salicornia (Davy et al. 2001). In Kadereit’s recent article (Kadereit et al.
2007), he concluded and summarized five main reasons why those taxonomic difficulties exist: lacking of diagnostic characters, inadequacy of dried herbarium material, misleading phenotypic plasticity, inbreeding and region treatment. Thus, the same widespread genotypes having been given different names in different regions while the same name was misapplicated of to different genotypes in one region. Besides, there still are uncertainties of delimitation of Salicornia and
Sarcocornia , both in subfamily Salicornioideae.
However, S. bigelovii , which is distributed in North
America, can be well distinguished from all other species of Salicornia by its acute and sharply mucronate leaf and bract tips according to Flora of North America (Ball
2003). It may be the breakthrough to wake from the taxonomic nightmare in method of chemotaxonomy. In the present work, we reported one new nortriterpenoid glycoside (3-O-[
β
-D- glucuronyl butyl ester]-30norolean-12, 20 (29)-dien-23-28-O-[
β
-D-glucopyranosyl] ester), together with three known nortriterpenoid glycosides (3-O-[
β
-D-glucuronopyranosyl]-30-norolean-
12, 20 (29)-dien-28-O-[
β
-D-glucopyranosyl]ester; 3-O-
[
β
-D-glucuronopyranosyl]-30-norolean-12, 20 (29)-dien-
23-aldehyde-28-O-[
β
-D-glucopyr anosyl]ester; 3-OH-30norolean-12, 20 (29)-dien-28-O-[
β
-Dglucopyranosyl]ester) from the whole plant of this morphologically distinct species cultivated in Jiangsu,
China. Previous phytochemical studies with Salicornia species showed mainly flavonoids. It is worth noting that the flavonoids were identified as the principal constituents of some species of this genus that occur mainly in the Europe and Asia. However, studies with
Salicornia species collected in Europe and America reported that sopains may occur (Glenn et al. 1991;
Lellau & Liebezeit 2001), and our study agreed with those researches. Although oleanolic acid is ubiquitous in the plant kingdom and of little chemotaxonomic significance, nortriterpenoid is rare and we consider it is important chemotaxonomic markers within the genus
Salicornia as well as may distinguish Salicornia from
Sarcocornia. References: Ball, WP 2003. Davy, AJ et al.
2001. Glenn, EP et al. 1991. Kadereit, G et al. 2007.
Lellau, TF & Liebezeit, G 2001.
P1051 – Poster
Phylogenetic relations of the South African species of
Caroxylon
(sections
Caroxylon
and
Tetragona
) based
1 on the morphology and nrITS sequence data
Feodorova, TA 1
Lomonosov Moscow State University, Moscow, Russia
Previous phylogenetic analyses have resolved 3 species of Caroxylon sect. Caroxylon as separate monophyletic group. The present study aims are to reconstruct phylogenetic relationships within the South African species of Caroxylon and Tetragona Ulbrich sections.
All studied species of Caroxylon and Tetragona from
South Africa form a monophyletic group and are derived.
The C. abarghuense (Irania sect.) from Iran and species from Belanthera section appear in basal position of South
African species group. The species C. humifusa
( Tetragona sect.) forms clade of late divergence of species of Caroxylon section. Supposedly, South African species ought to be viewed at as Caroxylon section sensu lato (type species C. aphyllum ), but basal North African and Asian species comparable to Irania and Tetragona
(type species C. tetragonum Delile from North Africa and Arabian peninsula) sections. The phylogenetic analysis indicates that the Caroxylon section is highly polyphyletic and weak support of other clades can testify about polyphyletic arising of Caroxylon genus and support a devision of the Caroxylon genus into several genera.
P1052 – Poster
First insights into the phylogeny of toadflaxes (
Linaria
Mill., Antirrhineae): systematic, evolutionary and biogeographic implications
Fernández-Mazuecos, M 1
MF 3 , Sáez, L 4 , Vargas, P 1
, Blanco, JL 1 , Juan, A 2 , Fay,
1 Real Jardín Botánico, CSIC, Spain;
Universidad de Alicante, Spain; 3
2 CIBIO,
Jodrell Laboratory,
Royal Botanic Gardens, Kew, UK; 4 Universitat
Autònoma de Barcelona, Barcelona, Spain
Toadflaxes ( Linaria Mill.) constitute the largest genus within the snapdragon lineage (tribe Antirrhineae,
Scrophulariaceae s.l.). This genus comprises nearly 150 species classified into seven sections: Linaria (45 species), Supinae (44 species), Versicolores (21 species),
Speciosae (19 species), Diffusae (17 species),
Macrocentrum (2 species) and Pelisserianae (2 species).
Two major groups have traditionally been recognized based on seed morphology: winged seeds (sections
Linaria , Supinae and Pelisserianae ) vs. wingless seeds
(sections Versicolores , Speciosae , Diffusae and
Macrocentrum ). The genus is mainly distributed in the
Mediterranean region (where it has its diversity centre) and Eurasia. Four seed-wingless species from North and
South America have been variably included within
Linaria as an additional section ( Lectoplectron ) or separated as a different genus ( Nuttallanthus D.A.
668

Sutton). Systematic, evolutionary and biogeographic hypotheses for Linaria have not been tested in a molecular phylogenetic framework to date. Here we present the first phylogenetic hypothesis of Linaria based on nuclear ribosomal DNA sequences of the Internal
Transcribed Spacer (ITS1-5.8S-ITS2). We performed a wide sampling of all sections and geographical regions, with special emphasis in the western Mediterranean species. Our results confirmed the naturalness of Linaria including the American ( Nuttallanthus ) species. The two major groups of wing- and wingless-seeded species were not supported, which indicates parallel evolution affecting this trait. Systematic implications regarding the naturalness of sections based on our results are discussed.
For example, the monophyly of sect. Supinae is supported, and three mayor groups are differentiated by floral sizes. These groups are in disagreement with previous morphological classification where the relationships within the section were established by differences in seed shape. Therefore, within sect. Supinae subsections Supinae and Saxatilis must be discarded as systematic units. In the case of sect. Versicolores , the analysis indicated monophyly for both the section and two subsections . In summary, certain disagreement between molecular phylogenies and morphological characters may be responsible for multiple taxonomic treatments published in the last century. In any case, our phylogeny confirms the Mediterranean region as the origin and main centre of diversification of Linaria lineages, and a Miocene origin for the genus is suggested by molecular-clock estimation. The biogeographic implications of the phylogenetic position of American and Asiatic species (sect. Lectoplectron and sect.
Linaria ) are also discussed.
P1053 – Poster
Genetic diversity of the native fruit Quandong
(
Santalum acuminatum
) in South Australia
, Gardner, M 2 , Cross, H 3 , Lowe, A 1
1
Fuentes-Cross, P 1
School of Earth and Environmental Sciences, University of Adelaide, Australia; 2 School of Biological Sciences,
Flinders University, Adelaide, Australia; 3 State
Herbarium of South Australia, Dept Environment and
Natural Resources, Australia
The Australian native hemiparasitic tree Quandong
( Santalum acuminatum ) is widely distributed along the southern dry regions of Australia. One of the best known bushfoods, the Quandong is facing increasing threats from wild harvesting for their fruit, browsing by feral animals, and habitat fragmentation. As the Quandong becomes more economically relevant, studies aimed at understanding its genetic diversity are lacking . In our present project, we use a phylogeographic approach to study remnant populations in South Australia, reconstruct the historical processes underlying their geographic distributions, reveal their current genetic structure, and determine which of the external stresses have had the greatest impact . We collected leaf samples from extant populations of Santalum species across South Australia and from the State Herbarium. These samples have been genotyped using chloroplast and nuclear microsatellite markers, and DNA sequences have been obtained from the nrDNA marker ITS. Preliminary results indicate a high degree of genetic variation both between and within populations of South Australia. This was found in DNA sequences of both ITS and chloroplast regions. Both
DNA sequence and microsatellite data suggest that populations from western South Australia, in the
Nullarbor region, were genetically distinct from all other populations sampled. This corresponds with morphological variation observed in these populations.
More detailed genetic and morphological study of
Quandong will help to uncover the historical processed underlying the current distribution of the species, and determine what biotic and abiotic factors have had the largest impacts on their genetic diversity.
P1054 – Poster
Phylogenetic relationships, character evolution and taxonomic reexamination of Nartheciaceae
(Dioscoreales)
Fuse, S 1 , Lee, NS 2 , Tamura, MN 3
1 Museum of Nature and Human Activities, Hyogo,
Japan; 2 Dept of Life Science, Ewha Womans University,
South Korea; 3 Dept of Botany, Graduate School of
Science, Kyoto University, Japan
Using DNA sequences of plastid atp Brbc L, trn L and trn Ltrn F as well as nuclear 18SrDNA (4048 bp in total) from 31 accessions, out of which 22 were sequenced by ourselves and nine obtained from the DNA Data Bank of
Japan (DDBJ), of 19 species of all the five genera of
Nartheciaceae (Liliaceae sensu lato), we conducted a combined analysis to contribute to the understanding of phylogenetic relationships, evolution of characters, and recognition of genera and species in Nartheciaceae.
According to strict consensus of 16 most parsimonious trees, two clades are formed in the family: one of
Metanarthecium and the other of the rest of the four genera, although the latter clade receives only 60% bootstrap support. In the latter clade, Aletris is diverged first, Lophiola is as the next branch, and then Nietneria and Narthecium form a clade (100%). The accession, which was used as a representative of Metanarthecium in an earlier study and led the study to the conclusion that
Metanarthecium was congeneric to Aletris , forms a clade not with other three accessions of Metanarthecium but with Aletris foliata in the present analysis. The accession of Metanarthecium in the earlier study might be correctly identified as Aletris foliata , which sometimes grows sympatrically with Metanarthecium . In the present study, contrary to the earlier study, Metanarthecium turned out to be independent from Aletris . As for findings at species level, Aletris fauriei ought to be reduced to A. foliata .
Nartheciaceae are famous for their variability of the characters, which are primarily considered to be important in taxonomy of the monocots, such as leaves unifacial vs. bifacial, septal nectaries presence vs. absence and ovary superior vs. half-inferior / inferior.
Based on the present molecular tree, a shift from bifacial leaves to unifacial ones as well as a loss of septal nectaries are presumed to have occurred once at the common ancestor of Lophiola , Nietneria and
Narthecium , respectively. On the other hand, a shift from superior ovary to half-inferior / inferior ovary is probably presumed to have occurred three times in the family, i.e. at the ancestor of Aletris , at the ancestor of Lophiola and
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at the ancestor of Nietneria . Further, we analyzed plastid trn K (including mat K) in addition to the regions used in the former analysis, i.e. atp Brbc L, trn L, trn Ltrn F and
18SrDNA, (6337 bp in total) from 21 accessions of 13 species of the Nartheciaceae, in order to confirm especially the clade of the nartheciaceous genera excluding Metanarthecium , although we were unable to include Nietneria in this analysis. The bootstrap support for the clade was 60% in the former analysis, but increased to 80% in the latter analysis. Thus, the probability of exclusion of Metanarthecium from Aletris turned out to be comparatively high.
P1055 – Poster
1
New reports of androdioecious species from Rutaceae
Fuse, S
Japan;
1
2
, Ushijima, K 2
Junior College, Japan
, Ushijima, T 2 , Kurosaki, N
The Society for Flora of Hyogo, Japan;
3
Museum of Nature and Human Activities, Hyogo,
3 Shoei
Zanthoxylum ailanthoides Siebold et Zucc. and Z. schinifolium Siebold et Zucc. are deciduous tree species distributed in East Asia. These species have been reported to be dioecious in many literatures. However, we observed monoecious individuals as well as male individuals from several natural populations in Hyogo
Pref., Japan, and we could not find any female individuals from the same populations. Thus, we investigated flowering phenology and pollen fertility of the individuals that constitute the unique populations of the two species in order to know the reproductive characteristics. And, we revealed the following results.
(1) Flowering periods of male flowers of male individuals are longer than those of male flowers of monoecious individuals. (2) The monoecious individuals are protandrous, i.e. male stage --> sterile stage --> female stage. (3) However the shifts of the stages are not synchronized among individuals in the same population.
Thus both the male stage and female stage are observed at the same time in each population. (4) As for both male flowers of male individuals and male flowers of monoecious individuals, pollen fertilities are more than
95%. These results indicate that Z. ailanthoides and Z. schinifolium are functionally androdioecious.
P1056 – Poster
Phylogenetic analysis of
Gollania
(Hypnaceae) using
1 plastid DNA sequence data
Higuchi, M 1 , Arikawa, T 2
2
National Museum of Nature and Science, Japan;
Tottori Prefectural Museum, Japan
The genus Gollania is a pleurocarpous moss of 20 species distributed from mainly temperate regions of East
Asia. The genus has not previously been the subject of phylogenetic analysis based on either morphological or molecular characters. Although a preliminary phylogenetic analysis using the chloroplast gene rbc L has suggested that Gollania is monophyletic except G. taxiphylloides and G. splendens , the species level relationship has not been resolved. Using DNA
670 sequences from the chloroplast genes ( rbc L, rps 4 and psa B) and more species samples, we are investing the phylogenetic relationships of Gollania . To test the position of the genus, a comprehensive analysis using a combined set of rbc L, rps 4 and psa B sequences were applied including more than 230 samples of over 220 hypnalean species. The analysis shows that
Macrothamnium is a sister group of Gollania . We included twelve species of Gollania and used two species of Macrothamnium as outgroup. A phylogenetic hypothesis based on chloroplast DNA data indicates that
Gollania is monophyletic except G. taxiphylloides and G. splendens . In Gollania clade, four main clades are recognized: (1) clade A ( G. japonica and G. setschwanica ), (2) clade B (G . turgens and an undescribed species), (3) clade C ( G. robusta ) and (4) clade D ( G. varians and the remaining species). Although interpretations of morphological evolution are constrained by the limited extent of species sampling, several morphological synapomorphies can be identified from specific clades. Species in clade A and B have a differentiation of stem and branch leaves, whereas species in clade C and D have no differentiation; two species of Macrothamnium have a differentiation of stem and branch leaves. The G. c ylindricarpa –undescribed species– philippinensis lineage within clade D possesses a synapomorphy for no differentiation of dorsal, lateral and ventral stem leaves; all other species in the phylogenetic analyses possess the differentiation.
P1057 – Poster
1
Can root adaptations explain Cape species richness?
Huber, M 1 , Linder, P 1
Institute of Systematic Botany, University of Zurich,
Switzerland
The species rich South African fynbos is characterized by a high turn-over in species composition and much of its diversity is structured along soil moisture gradients. Here we investigate how species of the Restionaceae (Poales) vary in their roots, and whether this variation can account for species segregation along a hydrological gradient. We found that aerenchyma – tissue with enlarged gas spaces to avoid anoxia during waterlogging – is nearly exclusively present in species characteristic of wetlands.
Cultivation experiments confirmed the waterlogging– aerenchyma relationship, as the ability to make this structure was highly correlated with the survival rate during submergence. Field observations showed that aerenchymatous plants were over-represented in waterlogged soils and under-represented in well-drained soils, suggesting ecological sorting according to soil drainage capacity. Phylogenetic analysis shows that aerenchyma is both phylogenetically constrained, and also co-evolved with impeded soil drainage. These results suggest that aerenchyma plays an important role in promoting species segregation along a moisture gradient and thus partially explains species richness in the Cape flora.
P1058 – Poster
Taxonomic implications of pollen morphology in infrageneric classification of
Scutellaria
(Lamiaceae)

Jamzad, Z 1 , Hasani-Nejad, M 1
1 Research Institute of Forests & Rangelands
Scutellaria L., subfamily Scutellarioideae is a subcosmopolitan genus with about 360 species. The latest infrageneric classification of Scutellaria and its allied genera, Salazaria Torrey, Perilomia Kunth, and
Harlanlewisia Epling, revealed close relationships among Scutellaria and the above mentioned genera and they were all included in Scutellaria . Two subgenera are recognized in the genus, characterized as follows: subgenus Scutellaria , with one-sided inflorescence in which flowers are subtended by leaves or leaf like bracts and subgenus Apeltanthus , with four-sided inflorescence and cucullate bracts. Five sections were recognized in the type subgenus, including sections Scutellaria , Perilomia
(Kunth) Epling emend. Paton, Salazaria (Torrey) Paton,
Anaspis (Rech.f.) Paton and Salviifoliae (Boiss.)
Edmondson. We studies pollen grains of 25 species of
Scutellaria from Old and New World to evaluate its taxonomic implication in infrageneric classification of the genus. The species sampled belong to subgenus
Scutellaria , sections Scutellaria , Anaspis , Perilomia ,
Salazaria and subgenus Apeltanthus (Nevski ex Juz.)
Juz. emend. Paton, section Lupulinaria A.Hamilton. The pollen grains examined were taken from collections in the Herbaria of the Harvard University (HUH) and
Research Institute of Forests and Rangelands (TARI), and were acetolysed and studied by SEM. All pollen grains that were examined were isopolar, tricolpate, suboblate, oblate spheroidal to euprolate. The exine was tectate-perforate, to microreticulate, with suprareticulate sculpturing. Supratectal sculpturing consisted of discontinuous ridges, or formed a supratectal reticulum.
In the examined species of subgenus Scutellaria sects.
Scutellaria , Perilomia and Salazaria supratectal sculpturing consisted of ridges forming supratectal reticulum; muri was angular, lumina had an irregular shape; the perforations were more or less uniform and of the same size. In sect. Anaspis supratectal ridges were discontinuous and the tectum uniformly perforated. In species of subgenus Apeltanthus sect. LupuLinaria exine was microreticulate, the lumina contain one or more large central perforations surrounded by small ones. This study thus reveals that pollen exine sculpturing is a useful taxonomic character for infrageneric classification of Scutellaria . Our results are congruent with the current classification of the genus proposed by Paton.
P1060 – Poster
Bird pollination syndromes in the Australian
Passiflora
(subgenus
Decaloba
: supersection
Disemma
)
Krosnick, S 1 , Miles, M 1
1 Dept of Biology, Southern Arkansas University,
Magnolia, Arkansas, USA
There are over 550 different species within the genus
Passiflora , most of which are found in Central and South
America. Some species may be herbaceous or shrub-like, but the majority of these species are vines or lianas.
Species of Passiflora have many different types of pollinators, the most common of which are bees, butterflies, and birds.
Of the three modes of pollination, bird pollination is the perhaps most interesting because it has evolved many times independently in the genus, and because in most cases very little is known about the details of these cases. The nectar content (glucose, sucrose, and fructose) differs from species to species, and is intricately connected to the type of pollinators associated with the flower. Flowers that are pollinated by hummingbirds or specialized nectar-feeding passerines will most likely secrete nectars rich in sucrose. Flowers that are pollinated by generalist nectar-feeding passerines, however, will most likely secrete nectars rich in glucose and fructose (hexose-rich) . The passerineassociated nectars are often very dilute, and may contain high levels of amino acids . This study focuses on three species of Passiflora endemic to the eastern coast of
Australia: P. cinnabarina, P. herbertiana , and P. aurantia . These species are especially interesting because they are part of a small lineage of Passiflora (24 species) that are native to the Old World. These three species are brightly colored red, orange, and yellow, and remain open for several days. They do not exhibit any floral scent, are held erect at anthesis, and have long androgynophores. Given these characteristics, it has been hypothesized that P. cinnabarina, P. herbertiana , and P. aurantia are bird pollinated. As no direct observation of pollinators have been documented for these species, the question remains as to what pollinators are associated with these flowers, and what type of nectar these species produce . As part of a visit to Australia in July 2011, field observations of pollinators and collection of nectar samples will be performed. This will be followed by
HPLC analysis to determine the sugar ratios for the nectar collected within each species. The results of a literature survey on ornithophilous pollination syndromes and nectar constituents in Passiflora are presented here to provide the necessary framework for interpretation of the data collected.
P1061 – ePoster
Architecture and inflorescence structure in epacrids
(Ericaceae: Styphelioideae).
1
Lemson, K 1
Centre for Ecosystem Management, Edith Cowan
University, Perth, Australia
Inflorescence characters have been used regularly in phylogenetic analyses of, or including, Styphelioideae; however their utility is reduced by problematic assumptions of homology and highly interpretational coding . Almost all species are polyaxial, woody shrubs or trees that flower repeatedly, generally on axes that are no different from 'vegetative' axes, and often without a clearly delineated 'inflorescence' . At the same time, there are also taxa (e.g. some Richea and Dracophyllum ) with highly complex, ephemeral structures that are morphologically differentiated from the rest of the plant .
As a consequence, it has been difficult to capture variation and describe homology relationships across the clade using traditional terms and concepts. An alternative approach exists that employs concepts developed in the field of plant architecture . Using units such as metamers, modules or shoot units and apical meristems as the points of comparison can provide a clear descriptive basis for
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direct comparisons of the underlying components of complex structures such as inflorescences, and facilitate their analysis from a whole-plant perspective . This is of import where the relationship between vegetative and reproductive growth is very close and meaningful analysis of putative homology relationships must consider flowering and non-flowering parts in an integrated fashion. This study examined a number of structural/architectural features in a suite of epacrids with widely differing inflorescences: the extremes of species with flowering axes that are almost indistinguishable from vegetative ones (e.g. Andersonia ), and a number with elaborate and highly differentiated inflorescences
(e.g. some Richea , Dracophyllum ), in addition to some intermediate forms (e.g. Astroloma ) . The taxa also included representatives from genera in which there is variation between species in the position of flowers
(exclusively axillary, exclusively terminal, or a mixture of both), and taxa from the various different clades currently recognised at tribal level. Plant structure was characterised in terms of metameric constitution, shoot unit construction, the timing of various developmental events (e.g. floral bud expansion) and the fate of the shoot apex. The findings provide a broadened basis for the formulation of characters for cladistic analysis, identifying a general similarity of structure among some groups of epacrids, but also underlining some significant differences. 'Inflorescences' occupy different levels within the hierarchy of repeating structural units in different taxa, making direct comparisons based on the traditional character 'flower position' complex or even meaningless. Needhamiella pumilio , Dracophyllum minimum, Sphenotoma spp. and members of tribe
Cosmelieae have similar patterns of shoot unit construction and growth, however other taxa differ in the positions of new vegetative growth and the orders of flower initiation and anthesis . Leaves widely termed
'bracts' and 'bracteoles' are not homologous across all taxa . Shoot dimorphism exists in Lysinema, Astroloma,
Epacris , and two species of Richea , and there is evidence for multiple axillary buds in Dracophyllum. Blastotely is universally associated with anauxotely, in contrast to some other Ericaceae in which architecture has been studied . The complex, multi-axis herbaceous inflorescences of Richea pandanifolia and a number of
Dracophyllum species are special cases worthy of further investigation.
P1062 – Poster
Promiscuous Proteaceae: natural hybridization between Australian
Lomatia
species
McIntosh, E 1 , Rossetto, M 2 , Wardle, G 1
1 University of Sydney, Australia; 2
, Weston, P 2
Botanic Gardens
Trust, Sydney, Australia
Natural hybridization events are important phenomena in the evolution of plant lineages yet many questions remain. Why do reproductive barriers separating closely related species break down in some places and not in others? How widespread is introgression, the movement of genes from one species into another? This study explores the role of natural hybridization the evolution of plant lineages, using the Australian native shrub species
Lomatia myricoides (Gaertner f.) Domin and L. silaifolia
672
(Smith) R.Br. (Proteaceae) as a model system . Lomatia myricoides (River Lomatia) is a tall shrub with long, linear leaves and indeterminate inflorescences, and is often found growing close to water sources. In contrast,
L. silaifolia (Crinkle bush) is a low shrub, with highly divided leaves and terminal inflorescences. Individual L.
myricoides x L. silaifolia hybrids can exhibit morphological traits from both parental species, and are extremely variable in terms of leaf shape, plant height and inflorescence structure. Hybrids occur at over twenty sites around the Sydney Basin and Blue Mountains area in eastern Australia where these species overlap at the edge of their distributional ranges . Three sites were closely analysed in this study, including a suspected hybrid swarm comprising of hundreds of individuals.
(Hybrid swarms are produced when crossing between two species is followed by backcrossing of hybrids with one or both parental species) . A combined approach of phenetic analyses of leaf and plant traits, genotyping using microsatellite markers, and records of flowering phenology were used to characterise patterns of variation within this hybrid complex . This diverse range of approaches also enabled exploration of the significance of hybridization as a process contributing to the adaptive potential of these species. In addition, we sought to determine which reproductive isolating barriers may have broken down to allow the formation of hybrid individuals, and whether morphological characters can be reliably used for the identification of hybrid individuals in the field . The genetic diversity and structure of hybrid populations was determined using fourteen microsatellite markers, enabling the complexity of hybridization events within Lomatia to be illucidated with New Hybrid software. Hybrids were classified as F1 (first filial generation), F2, or subsequent generations, enabling quantification of the degree of backcrossing and introgression occurring where hybrid swarms have developed . In an attempt to determine whether genetic differentiation mirrored the observed phenotypic variation present, morphological features such as inflorescence traits, plant height, and leaf shape characteristics were included in principle coordinate analyses and compared to genetic groupings .
Interestingly, hybrids were found able to reproduce, and hybrid seeds were observed to germinate as successfully as those of L.
myricoides. This project contributes to an ongoing investigation entitled ‘Speciation in the
Australian flora: testing explanatory hypotheses in waratahs and their allies’ which is currently being undertaken at the National Herbarium of New South
Wales. Understanding the processes shaping hybridization events in Lomatia will contribute to current debates about species boundaries and our understanding of speciation in the Australian Proteaceae.
P1063 – Poster
Chromosome numbers in
Arisaema
(Araceae) section odorata
J. Murata
Murata, J 1 , Ohi-Toma, T 1
1 Botanical Gardens, University of Tokyo, Japan
Arisaema sect. Odorata was distinguished from the sect.
Tortuosa of traditional circumscription by Gusman and
Gusman (2006) namely 'sect. Attenuata ' on the basis of

recent molecular phylogeny. According to the oldest lectotypification for section Attenuata , however, sect.
Attenuata should be a synonym of sect. Fimbriata and a new name becomes necessary for the group in question.
Sect. Odorata consists of the following 8 species
(chromosome numbers in parenthesis) in the sense of
Flora of China (2010): A. saxatile, A. aridum (2n=16), A. odoratum (22), A. mairei (22), A. lidaense (24), A. quinquelobatum (24), A. yunnanense (48), A. prazeri
(26). Arisaema aridum has been revealed to have the lowest chromosome numbers in Arisaema (previously 2n
= 20 in A. costatum was lowest). Because 2n = 16 and 22 are unique in Arisaema chromosome number might be reduced within this group.
P1064 – Poster
Diplosporous apomixis in
Hieracium
subgenus
Hieracium
(Asteraceae)
Siddons, HR 2 , Oelkers, K 2
Koltunow, AMG 1,2
, Johnson, SD 2 , Fehrer, J 3 ,
1
2
Corresponding Author: Anna.Koltunow@csiro.Au;
3
Division Plant Industry, CSIRO, South Australia;
Institute of Botany, Czech Academy of Sciences,
Pruhonice, Czech Republic
Reproductive development in the polyploid genus of
Hieracium , family Asteraceae, is unique in the plant kingdom; with two modes of asexual reproduction
(aposporous and diplosporous apomixis) coexisting with sexual development. To date, apomixis research has focused on aposporous Hieracium subgenus Pilosella ; while the reported diplosporous subgenus Hieracium has received less attention. Therefore we have analysed the previously uncharacterised mechanism of diplosporous apomixis in Hieracium subgenus Hieracium .
Reproductive analysis of 15 diplosporous and 6 sexual species of Hieracium subgenus Hieracium , revealed that the reported diplosporous subgenus Hieracium undergo mitotic diplospory; with no observation of meiotic divisions, as occurs in meiotic diplosporous Taraxacum .
Ovules then proceeded rapidly through autonomous seed development. Callose deposition was observed to be present capping the MMC within the early diplosporous subgenus Hieracium ovules, however at reduced levels compared to the sexual subgenus Hieracium and aposporous subgenus Pilosella . Meiotic processes, such as the visualisation of tetrads and functional megaspore selection were never observed within diplosporous subgenus Hieracium . Analysis of subgenus Hieracium determined that none of the diplosporous plants contained markers linked to the aposporous LOA (LOSS
OF APOMEIOSIS) locus of subgenus Pilosella .
Presence of callose capping the MMC within diplosporous ovules indicates that the MMCs are likely to have sexual identity at the start of reproductive development; with the MMC later being diverted into the mitotic diplosporous pathway before any meiotic divisional processes can occur. The absence of the conserved aposporous LOA markers is a likely indication that mitotic diplospory, as observed in subgenus
Hieracium , is likely to be controlled by a locus unrelated to the LOA locus of subgenus Pilosella .
P1065 – Poster
Origin and diversification of
Leucobryum boninense
(Leucobryaceae, Musci), endemic to the Bonin Islands
Emiko, O 1 , Tomio, Y 2 , Hiromi, T 3 , Hironori, D 2
1 Makino Herbarium, Tokyo Metropolitan University,
Japan; 2 Dept of Biological Science, Graduate School of
Science, Hiroshima University, Japan; 3 Miyajima
Natural Botanical Garden, Graduate School of Science,
Hiroshima University, Japan
Bryophyte species tend to have broad geographical ranges with a morphological uniformity in comparison with those of seed plants . Resent population genetic studies on bryophytes have revealed that some bryophyte species display high levels of genetic diversity and complex genetic structure . Although bryophyte species have the capability of long-distance dispersal, extreme geographical isolation may affect diversification and speciation . Thus, oceanic islands that have never been connected to any continental landmass may provide models for the study of diversification and speciation.
The Bonin Islands are typical oceanic islands . Located in the western part of the North Pacific, ca. 1,000 km south of Tokyo, Japan, they consist of 24 islands . The islands are of Eocene origin and appeared around the equator about 3 million years ago . The percentage of endemic vascular plants in the Bonin Islands is estimated at 40-
43% . Most of these endemic species do not have their closest allies within the islands, but rather in surrounding continental regions such as Southeast Asia and Oceania .
This suggests that geographical isolation in the Bonin
Islands as been sufficient to cause allopatric speciation of seed plants. A total of 155 species of bryophytes (48 genera and 81 species of mosses, 33 genera and 74 species of liverworts and hornworts) are currently known from the Bonin Islands . However, there remains much controversy about their source. Leucobryum boninense
Sull & Lesq. is an endemic to the Bonin Islands, and characterized by the perichaetia terminal on short lateral branches, and the proration on the abaxial surface of apical parts of the leaves . Related species, such as L. scabrum , L. javense var.
javense and L. scaberulum , are widely distributed in East and South East Asia . L. scaberulum , known from Hong Kong, Taiwan and
Okinawa Island, was treated as a synonym of L . boninense by Yamaguchi (1993) . To understand the evolutional history of L. boninense , phylogenetic relationships among L. boninens and its related species in the northwestern Pacific were analyzed using nuclear and plastid DNA. The analysis showed that L. boninense was a monophyletic, and formed two groups: Ogasawara
Islands (Chichijima, Hahajima and Anijima Islands) and
Iwo Islands (Kita-iwo island) . This species was closely related to L. javense var. javense from East Asia . These two species are distinguished from each other by plant size and the ratio of the length of the prorate apex region to the whole leaf length . Molecular and morphological evidence strongly suggests that L. boninense is endemic to the Bonin Islands, and may have originated from East
Asia . These results also suggested that there might be no barrier to gene flow via spore dispersal between islands and allopatric speciation has occurred in the genus
Leucobryum.
673

P1066 – Poster
GrassPortal: large-scale data synthesis offers a novel perspective on global diversity
Osborne, C 1 , Visser, Vernon
Chapman, Sam 3 , Freckleton, Rob
1
1
, Barker, Abigail
, Salamin, Nicolas 4
2
Simpson, David 2
Development Team 5
, Uren, Victoria 1 ,
,
,
GrassPortal
1 University of Sheffield, UK;
Kew, UK; 3
2 Royal Botanic Gardens,
Knowledge-Now Limited, Sheffield, UK;
4 University of Lausanne, Switzerland; 5 Various
Biological disciplines spanning molecular, evolutionary, ecological, ecosystem and Earth System sciences are accumulating data at unprecedented rates. Synthesis of these diverse data types at the interface between disciplines offers tremendous potential for major scientific breakthroughs. However, the rapid pace at which the information is being generated presents a major technical challenge to data aggregation, necessitating the development of automated mechanisms to forge scientifically rigorous links between disparate data resources. Here, for the first time, we use computer science methodology to automate the aggregation of taxonomic, phylogenetic, biogeographic and environmental data within a public web resource called
GrassPortal (www.grassportal.org). This focuses on the grass family (Poaceae), because of its species richness
(11,088 known species), worldwide distribution, major ecological and economic importance, and wealth of data availability. GrassPortal synthesizes information from four data resources: i) the GrassBase taxonomic database, which recognizes 59,380 synonyms, and includes expert range maps and information on morphological traits; ii)
GrassWeb, which provides phylogenetic relationships that are based upon molecular markers sequenced for
>3,000 species; iii) GBIF, which records geographical species occurrences, and contains >6.5 million records; and iv) multiple Earth Observation data products, describing the terrestrial environment. The major technical challenges that we have solved in this work include the cross-referencing and error checking of species names across multiple databases, the design of pipelines for automatic updates, and the construction of a flexible system that allows a broad range of possible queries. We illustrate two key products of our synthesis, showing: a) how global species richness and phylodiversity are distributed and sampled; and b) how biological traits sort along environmental gradients. By carrying out technically challenging bioinformatic data processing, our resource opens up new possibilities for the biological community, allowing a renewed focus on posing, refining and testing synthetic hypotheses about speciation, trait evolution and species coexistence.
P1067 – Poster
Changes recorded from 1970 through 2007 in base of the use of land and vegetation geographic data of
INEGI with 1:250,000 scale reference, México
Rodriguez-Avalos, JA
Alcocer, M 1
1 , Victoria-Hernandez, A 1 , Niño-
1 Instituto Nacional de Estadistica Y Geografia (Inegi),
Mexico
In México, the National Institute of Statistics and
Geography (INEGI) focuses on the generation of national mapping information of use of the soil and vegetation to scale average 1: 250,000 by its usefulness in regional and national studies currently boasting 4 time series: Series I,
II, III and VI (80's, 1993, 2002 and 2007 respectively).
The methodology for each has varied based on the available technology, which has evolved from opticmechanical devices and analog methodologies to digital platforms and processes semi- automated; however, their cartographic features allow various calculations and analyze the causes of the changes. In the process are calculated change polygons and transition type is valued to determine whether changes by use or methodological adjustments. The analysis in turn can be visualized by ecological provinces, regions hydrological or by national information classes. So the modification of natural vegetation cover shows a relative slowdown, but maintains a rate of about 400 thousand hectares per year, being the most affected communities grasslands, arid and semi-arid scrublands and deciduous forest. In contrast, forest and rain forest show a marked tendency to degrade towards secondary conditions rather than reduction in surface. Certain parts of the country show an incipient recovery by abandoned agricultural and livestock areas detected as secondary shrub vegetation. In general we see that change is still in favor of agricultural expansion, even though it has varied in intensity and is this focus on revive abandoned livestock lands and vegetation previously degraded rather than affect areas with preserved vegetation. In the coming decades exchange rate reduction could observe, however it also expect a significant drop in the quality of natural plant communities
P1068 – Poster
Distribution and description of the plant communities in Aguascalientes, Mexico
Rodriguez-Avalos, JA 1 , Siqueiros-Delgado, ME 2
Martinez-Ramirez, J 2
1
,
Instituto Nacional de Estadistica Y Geografia (Inegi),
Mexico; 2 Universidad Autonoma de Aguascalientes
(Uaa), Mexico
Aguascalientes is a small state located in the northerncentral part of Mexico. It houses a significant diversity of plant communities because of several ecological regions converged here, there are temperate forests from the
Sierra Madre Occidental, desert scrubs from the South-
West of the Chihuahua Desert, grassland from the
Durango–Zacatecas Highlands, tropical vegetation of the
Juchipila and Huejucar Canyons, and several temperate and subtropical communities from Los Altos of Jalisco.
In order to learn more about the current condition of the vegetation of Aguascalientes, the Autonomous
University of Aguascalientes (UAA) and the INEGI
(Instituto Nacional de Estadística y Geografía), are conducting a detailed survey to describe the composition, distribution, degree of conservation and degradation of the plant communities, currently present in
Aguascalientes. For this purpose, conventional
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vegetation sampling methods and digital geographic information techniques are used; we will evaluate dominance, abundance, covertures, frequency, plant diversity and floristic information. We also will circumscribe the distribution of plant communities through remote sensing techniques and fieldwork. At the end of the project, we will obtain 1:50 000 vegetation maps, and we will publish a book about the vegetation of
Aguascalientes. Until now, we got an advance of 50% in the fieldwork and data analysis; we proposed the first draft of the classification of plant communities of
Aguascalientes, and we recognized 42 variants of plant associations for the state. We have been able to corroborate the existence of various relictic areas, which evidenced significant changes as a result of the intensive exploitation of natural resources over the past century.
P1069 – Poster
Interactive identification and database for the
Myrtaceae tribe Chamelaucieae
Rye, B 1 , Trudgen, M 1 , Macfarlane, T 2 , Lander, N 1
1 Western Australian Herbarium, Dept of Environment and Conservation, Australia; 2 Western Australian
Herbarium, Dept of Environment and Conservation,
Australia
The Chamelaucieae is one of the largest tribes of
Myrtaceae, extending from southern China to Tasmania and with the great majority of its species Australian. A major concentration of genera and species occurs in the south-west of Western Australia, one of the world's biodiversity hotspots . With generic boundaries still very unclear in much of the tribe and many of the species lacking formal names, the identification of the western taxa is extremely difficult and there is no ready means for dissemination of new information. As a solution to these problems, an interactive key has been developed for the whole tribe, in conjunction with taxonomic revision.
Distribution data, flowering times and 155 morphological characters are used to discriminate between more than
760 taxa, with additional notes where these characters may be insufficient. Chromosome numbers, synonyms and common names are also listed and illustrations provided where possible. The key is intended to be maintained with regular updates made available through
Web delivery.
P1070 – ePoster
Molecular phylogeny of tribes Lithospermeae and
Boragineae with an emphasis on the genus
Onosma
(Boraginaceae s. str.) based on nrDNA ITS sequences in Iran
Saadati, N 1 , Kazempour Osaloo, S 1 , Maassoumi, AA 2
1 Dept of Plant Biology, Faculty of Biological Sciences,
2 Tarbiat Modares University, Tehran, Iran; Dept of
Botany, Research Institute of Forests and Rangelands,
Tehran, Iran
Boraginaceae s. str. (=subfamily Boraginoideae) comprises some 1600 species in approximately 100 genera and have their center of diversity in Eurasia. In
Flore of Iran , the family comprises 36 genera and over
180 species. The tribe Lithospermeae includes nine genera and 60 species and Boragineae includes eight genera and 20 species in Iran, being two of six traditionally recognized tribes. Onosma , as a member of
Lithospermeae, is the largest genus in the family in our area. 39 species of Lithospermeae (including eleven
Onosma species) along with 14 species of Boragineae and four species of the tribe Echieae , three species of the tribe Cerintheae and 23 species of Eritrichieae as well as two Echiochilon species plus Heliotropium bacciferum and Tournefortia rubicunda as outgroups were include in the phylogenetic analyses using nrDNA ITS sequence data. This region was amplified using appropriate primers by PCR and then PCR products were sequenced by the BigDye terminator cycle sequencing kit in an ABI
Prism 3730xl DNA Analyzer. The sequences were aligned using Muscle program. The phylogenetic reconstruction was conducted using maximum parsimony, maximum likelihood and Bayesian methods as implemented in PAUP*, Tree-Puzzle/GARLI and
Mrbayes softwares, respectively. The analyses showed that Lithospermeae with the inclusion of Cerinteae and
Echieae formed a well supported monophyletic group.
Thus, in agreement with previous works, these two latter tribes should be merged in Lithospermeae. The present data supported the monophyly of Onosma s. str., but its
Iranian members did not form a single clade. This was caused by the sister group relationship of Onosma rostellatum (belonging to the monotypic section
Podonosma ) with the European species. Likewise,
Onosma sect. Onosma was not monophyletic.
Podonosma orientale (= O. orientale ) was well allied with Alkanna species. The analysis revealed that Arnebia and Moltkia each separately formed a clade. Alkana and
Moltkiopsis are successive sisters to Cerinthe ,
Lithospermum and Buglossoides . The relationship of the monotypic genus Huynhia ( Huynhia pulchra ) found in
Iran is unresolved. Echiochilon with two analyzed species positioned at the base of the tree as sister to the
Lithospermeae clade. Our analyses also showed that tribe
Boragineae at the current status is not monophyletic, since Suchtelenia calycina was nested in Heterocaryum clade (Erithricieae). Other members of Boragineae including Symphytum , Nonea and five genera from
Brunnera through Anchusa , formed a trichotomy .
P1071 – ePoster
The New Zealand genus
Simplicia
, and a new genus endemic to Japan, two new members of subtribe
Poinae (Poaceae)
Soreng, RJ 1
H 3
, Gillespie, LJ
, Smissen, R 4
2 , Boudko, K
, de Lange, PJ 5
2 , Bull, R 2 , Koba,
1 National Museum of Natural History, Smithsonian
Institution, Washington DC, USA;
Nature, Ottawa, Canada;
2 Canadian Museum of
3 Dept of Natural Sciences, JF
Oberlin University, Machida, Tokyo, Japan; 4 Allan
5
Herbarium, Landcare Research, Lincoln, New Zealand;
Dept of Conservation, Auckland, New Zealand
Simplicia and a new genus separated from Poa are newly placed in the grass subtribe Poinae (Poaceae, tribe
Poeae) . Phylogenetic analyses of plastid ( trn Ttrn Ltrn F) and nuclear ribosomal (ITS and ETS) DNA resolve both
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genera in a clade including all subtribe Poinae members except Poa. Simplicia is a ditypic genus endemic to New
Zealand with laterally compressed, glabrous, single flowered spikelets and glumes distinctly shorter than the lemma it . The species are confined to forested areas with
S. laxa favouring lowland to montane forest and S. buchananii montane to subalpine forest ( S. laxa also persists in the deforested landscape of Central Otago,
South Island where it grows in rock overhangs and cave entrances). The genus was described in 1897, but as both species are extremely uncommon (both are threatened) the genus and the species remained poorly known until it was fully treated in 1971. Simplicia was previously allied with Calamagrostis (subtribe Agrostidinae) or suggested to be close to Aniselytron (sometimes included in
Calamagrostis , but recently realigned in subtribe Poeae) .
Simplicia resolves with the south-east Asian genus
Aniselytron in nuclear ribosomal trees, supporting the previous suggestion of their close relationship . We provide evidence that Poa hayachinensis represents a new monotypic genus endemic to alpine limestone habitats in Hokkaido and northern Honshu islands of
Japan. The species forms a strongly supported clade with two circumarctic genera (absent from Japan), Arctophila and Dupontia , and resembles those genera in several flowered spikelets, lemmas that are frequently mucronate, sometimes 3-veined and with long scareoushyaline apices, and calluses with a crown of hairs around the base of the lemma . Poa hayachinensis differs from these two genera in having glumes shorter than the lowest floret, densely scabrous palea keels, and ciliate tipped lodicules, and from Poa in lemmas principally 3veined, distally golden-brown scareous to hyaline for
1/3–1/4 the length, frequently with an apical mucro, and ciliate tipped lodicules .
P1072 – Poster
Another candidate for the sister group of the monocotyledons
Tamura, MN 1 , Yamashita, J 2 , Fuse, S 3
1 Dept of Botany, Graduate School of Science, Kyoto
2 University, Japan; Institute of Plant Science and
Resources, Okayama University, Japan; 3 Museum of
Nature and Human Activities, Hyogo, Japan
These days, eudicots are often considered as the sister group of the monocots, based on the molecular phylogenetic study, in which very long plastid DNA sequences of 76583 bp were compared. However, this result was yielded by analyzing many morphologically advanced taxa rather than primitive ones as representatives of the monocots. In the analysis, nine materials of commelinids, including six materials of
Poaceae, out of 14 monocot materials were used, while
Alismatales, Petrosaviales, Pandanales and Liliales were not used at all as materials. Thus, we try to ascertain whether the sister group of the monocots is eudicots even if many morphologically primitive taxa of the monocots instead of advanced ones are used in a molecular analysis. In the present study, we analyze at least one taxon from each order of the monocots in addition to the morphologically primitive monocots, comparing DNA sequences of about 20000 bp, which comprise atp B, rbc L, mat K, ndh F, rpl 2 and rps 4 from plastid genome,
676 atp A, atp 1, cob, cox1, cox3, mat R and nad 5 from mitochondrial genome, and nuclear 18SrDNA. Our analysis has not been finished yet, but based on the interim results at the present stage, the sister group of the monocots might not be eudicots but Piperales.
P1073 – Poster
Origin and genetic structure of
Zostera japonica
(Zosteraceae), a seagrass distributed over a wide water-temperature range
1
Tanaka, N 1
M 4
, Ito, Y
National Museum of Nature and Science;
West Hungary; 3 Fisheries Research Agency;
University, Japan
2 , Hirayama, Y 1 , Saitoh, K
2
3 , Nakaoka,
University of
4 Hokkaido
Zostera japonica (Zosteraceae) is distributed from the intertidal zone to the subtidal zone, mainly within the
Japanese Archipelago, from Sakhalin to Northern
Vietnam. It has a wide water–temperature range as well as a wide variation in leaf morphology; therefore, some subspecies and ecotypes have been described within this species. Previous molecular phylogenetic studies of
Zosteraceae have revealed that Z. japonica is a sister group of the European species Z. noltii . In the present study, phylogenetic analysis was performed to detect the historical genetic disjunction in these two species.
Haplotype analysis using plastid and nuclear DNA and population genetic analysis with microsatellite loci were performed in 60 populations of Z. japonica obtained from the distribution area. Phylogenetic analysis revealed that the ancestral taxon of Z. japonica , Z. noltii , diverged into two phylogenetic groups: one, comprising Z. noltii and a part of Z. japonica ( Japonica -1); and the other comprising Z. japonica ( Japonica -2) alone. Z. noltii is an ingroup of Z. japonica , and which is a paraphyletic group. Haplotype analysis showed that Japonica -1 is located northward from central Japan and Japonica-2 is located southward. The distribution of the two groups overlaps around central Japan where individuals having both haplotypes were detected. Therefore, the possibility of reproductive isolation by genetic factors is absent.
Microsatellite analysis showed that sea current and coastal topography mainly influenced seed dispersal within southward and northward area respectively. We discuss environmental factors related to the distinctive distribution of the two groups of Z. japonica .
P1074 – Poster
West to east dispersal and subsequent rapid diversification of the mega-diverse genus
Begonia
(Begoniaceae) in the Malesian Archipelago
Thomas, DC 1 ,
Richardson, JE 2
Hughes, M 2 , Twyford, AD 2 , Ardi, WH 3 ,
2
1 University of Hong Kong, Hong Kong, PR China;
Royal Botanic Garden Edinburgh, Edinburgh, UK;
3 Bogor Botanic Gardens, Bogor, Indonesia
The complex palaeogeography of the Malesian
Archipelago, characterised by the evolution of an everchanging mosaic of terrestrial and marine areas

throughout the Cenozoic, provides the geographical backdrop for the remarkable diversification of Malesian
Begonia (>450 species). This study aimed to investigate the origin of Malesian Begonia , the directionality of dispersal events within the Malesian Archipelago, the impact of ancient water gaps such as the Makassar and
Lombok Straits on colonisation patterns, and identify potential drivers of diversification. Plastid DNA sequence data of representatives of all families of the
Cucurbitales and Fagales ( mat K, rbc L, trn L intron, trn L-
F spacer, 4076 aligned positions, 92 taxa) and a sample of all major Asian Begonia sections (ndhA intron, ndhFrpl 32 spacer, rpl 32trn L spacer, 4059 aligned positions,
112 taxa) were analysed under an uncorrelated-rates relaxed molecular clock model to estimate the age of the
Begonia crown group divergence and divergence ages within Asian Begonia . Ancestral areas were reconstructed using a likelihood approach implementing a dispersal-extinction-cladogenesis model, and with a
Bayesian approach to dispersal–vicariance analysis. The results indicate an initial diversification of Asian Begonia in continental Asia in the Miocene, and subsequent colonization of Malesia by multiple lineages. There is support for at least five independent dispersal events from western Malesia to Wallacea dating from the Late
Miocene to the Pleistocene. Begonia section
Petermannia (>270 species) originated in Western
Malesia, and subsequently dispersed to Wallacea, New
Guinea and the Philippines. Lineages within this section diversified rapidly since the Pliocene coinciding with rapid orogenesis on Sulawesi and New Guinea. The predominant directional trend of Begonia dispersals between continental Asia and Malesia, and within
Malesia has been from west to east. The water bodies which have separated the Sunda Shelf region from
Wallacea have been porous barriers to dispersal in
Begonia once substantial land had emerged in eastern
Malesia from the late Miocene onwards. We hypothesise that the formation of topographical heterogeneity and the promotion of microallopatry by orogenesis in the Plio- and Pleistocene, as well as cyclic vicariance by frequent habitat fragmentations and amalgamations because of climate and sea-level fluctuations during the Quaternary have been major drivers of the diversification of Begonia in Malesia.
P1075 – Poster
Flora of Australia Online – where to now?
Thompson, H 1 , Wilson, A 1 , Preece, M 1
1 Australian Biological Resources Study (Dept of
Sustainability, Environment, Water, Population and
Communities), Australia
The Flora of Australia series is progressively being databased. The Flora of Australia online website is http://www.environment.gov.au/biodiversity/abrs/onlineresources/flora/main/inde x.html
. The database currently delivers the material from the published book with some updates. The first volume of the Flora of Australia was published in 1982 and has not been updated . The database will link dynamically to the Australian Plant
Census for nomenclatural information, the Australian
Plant Image Index for additional images, the Australia's
Virtual Herbarium for maps, and pdfs of the original
Flora of Australia publication. Changes and new names will be added to keep the information current . As part of the Atlas of Living Australia project, improved accessibility and a public editing interface with a registered logon are being developed for the Flora of
Australia Online.
P1076 – Poster
The structure of the pollen presenter in selected southern African members of the tribe Vanguerieae
(Rubiaceae)
1
Tilney, PM 1 , van der Merwe, CF 2 , van Wyk, AE 2
University of Johannesburg, Johannesburg, South
Africa; 2 University of Pretoria, Pretoria, South Africa
Secondary pollen presentation is a common phenomenon in the Rubiaceae. Particularly conspicuous pollen presenters are found in the tribe Vanguerieae. These structures are knob-like and are formed by a modification of the upper portion of the style and stigma. While in the bud stage, the anthers surround the pollen presenter and release pollen which adheres particularly to longitudinal grooves in the pollen presenter. The epidermal cells facing the anthers are elongated with characteristic wall thickenings towards the distal end and frequently also proximally. In this study, the pollen presenters in selected southern African members of the tribe
(representing the genera Afrocanthium, Canthium ,
Keetia, Pachystigma, Plectroniella, Psydrax,
Pygmaeothamnus and Vangueria ) were studied using light microscopy. Comparisons were made to ascertain the taxonomic potential of these structures at the generic and, in some cases, also the species levels. In addition, a
TEM study was carried out on parts of the pollen presenter, with special attention being given to the distal and proximal cell wall thickenings associated with some of the epidermal cells. The epidermal cells have a particularly intricate ultrastructure with an abundance of organelles, including microtubules, endoplasmic reticulum and mitochondria being associated with the thickenings. This suggests an active physiological role for these cells rather than mere physical support for the adherent pollen grains as has hitherto been assumed.
Suggestions are made as to the possible functional significance of the various tissues and cellular structures in the pollen presenter.
P1077 – Poster
Phenetic similarity of Alismatidae subclass fruits
Toma, C 1
1 Pomeranian University, Poland
In total, 360 fruits of 12 species of the subclass
Alismatidae, collected in Poland (Europe) in the years
1996-2005, were studied. The aims of this researches included: 1) recognition of diagnostic features of mature
Alismatidae fruits, 2) determination of connections between the studied taxa of Alismatidae, based on morphological and anatomical characteristics of fruits and seeds, 3) preparation of a carpological key for identification of the genera of Alismatidae . The
677

following laboratory techniques were used: paraffin method, microtome sectioning, dying with safranine and fast green, preparation of permanent microscopic slides of transverse sections of fruits and the SEM studies with the help of the Hitachi S-3400N and S-3500N microscopes. The SEM images were analyzed with the image analyzer Met-Ilo 8 software and statistical analyses were conducted with the Statistica 7.1 software.
Statistical methods included: a cluster analysis related to the cases and features, multidimensional scaling and a dendrite method. Taxonomical recapitulation: 1) examined fruits were identified and classified into fruit types, 2) on the basis of morphological and anatomical fruit analyses 47 taxonomically useful diagnostic features have been selected, 3) carpological keys for identification of Alismatidae fruits were worked out, 4) phenetic similarity connections between the taxa of Alismatidae were established with the use of a cluster method, multidimensional scaling and a dendrite method; all these methods delivered very similar results of phenetic similarity of fruits in this group of plants. Based on morphological and anatomical features of fruits and seeds, the following groups among the Polish genera of
Alismatidae were distinguished: group 1 – with
Hydrocharis and Stratiotes , group II – Butomus , group III
– Scheuchzeria , group IV – Triglochin , group V – Alisma ,
Sagittaria , Luronium and Najas and group VI –
Potamogeton , Zannichellia and Ruppia . The smallest phenetic similarity of fruits was found between group I and II and the rest of groups and between group III and
IV. Higher phenetic similarity show groups V and VI .
P1078 – Poster
Phylogeny and taxonomy of
Liparis section
Liparis
in
Japan
Tsutsumi, C
Yang, CK 4
1 , Yukawa, T 1
, Kato, M 1
Science; Dept of Life Science, Ewha Womans
University, South Korea; 3
, Lee, CS 2 , Lee, NS 2 , Jin, X 3 ,
1 Dept of Botany, National Museum of Nature and
2
Institute of Botany, Chinese
Academy of Sciences, China; 4 The Experimental Forest,
College of Bioresources and Agriculture, National
Taiwan University, Taiwan
Liparis (Orchidaceae) consists of ca. 400 species and is distributed worldwide. In section Liparis , one of 19 sections, 9 of 15 species may be assigned to sect. Liparis in Japan. The classifications, however, are controversial, and the morphology-based taxonomic treatments of plants called as L. japonica, L.
kumokiri, L. makinoana and relatives differ with botanists. To clarify their taxonomy and systematics, we conducted a molecular phylogenentic analysis using samples of Liparis sect.
Liparis collected from Japan, China, Korea and Russia.
Molecular phylogenetic trees deduced from several chloroplast and nuclear regions suggest that plants called
L. japonica, L. kumokiri, L. makinoana and relatives are separated into two clades; one clade consists of L. japonica, L. makinoana and the relatives (called the
Makinoana clade), and the other clade comprises L. kumokiri and the relatives (called the Kumokiri clade). In the Makinoana clade, although at least three morphological types are distinguishable in L.
makinoana and L. japonica in Japan, their genetic differentiations are
678 subtle. In the Kumokiri clade, L.
campylostalyx, L. kumokiri, L. purpureovittata, L. koreojaponica, L. fujisanensis, and L. pterosepala are recognized morphologically and phylogenetically. Based on the results of comparative morphology and molecular analysis, we propose a new taxonomy and systematics of
Liparis sect. Liparis in Japan.
P1099 – ePoster
Fluorochrome chromosome banding of three
Adenocalymma
species (Bignoniaceae) from Brazilian
Cerrado
Brogliato Junqueira Engel, T 1 , Yamagishi-Costa, J 1
1
Salles Sampaio, D 1
Universidade Federal de Uberlândia, Brazil
,
The Brazilian Cerrado, currently with about 438910km²
(only 21.6% of its original area), is an important hotspot, continually and constantly threatened by the rapid expansion of agricultural activities. This alarming fact highlights the importance of studying and understanding the species of Cerrado to define and improve conservation strategies. Cytogenetic studies allow us to analyze the morphology, organization, replication, variation and function of chromosomes, and are therefore widely used in order to elucidate phylogenetic relationships, intraspecific and interspecific polymorphisms and chromosomal evolution.
Fluorochrome banding makes possible a thorough analysis of structure and possible changes in plant chromosomes, because the most commonly used dyes,
DAPI (4'6-diamidino-2-phenylindol) and CMA
(chromomycin A3), show affinity for AT and GC-rich
DNA regions, respectively, usually representing highly repetitive heterochromatic blocks. Adenocalymma campicola (Pilg.) L.G.Lohmann, A. pedunculatum (Vell.)
L.G. Lohmann and A. peregrinum (Miers) L.G.Lohmann are subshrubs of Bignonieae tribe, belonging to the
Bignoniaceae family and have 2n=40 chromosomes. The aim of this study was to determine the pattern of CMA and DAPI bands for A. campicola, A. pedunculatum and
A. peregrinum chromosomes, in order to help untangling and elucidating phylogenetic relationships and chromosomal evolution of species. Fruits of A. campicola (Caldas Novas, GO), A. pedunculatum
(Brasília, DF) and A. peregrinum (Uberlândia, MG) were collected and seeds were germinated at room temperature
(RT). Root tips were pre-treated with anti-mitotic (PDB,
4 hours at 16°C or 8-Hq, 24h at RT), and then fixed in
Carnoy solution (ethanol 3:1 acetic acid). Meristems were digested in an enzymatic solution containing cellulase and pectinase (2:20) and slides were prepared using traditional squashing technique. Coverslips were removed after immersion in liquid nitrogen and slides aged 3 days before sequential staining with CMA and
DAPI. Slides were analyzed and images captured under epifluorescent photomicroscope . A. campicola presented three CMA+ bands, two corresponding to secondary constrictions and a smaller telomeric, representing an heteromorphic pair of chromosomes, and no DAPI+ bands. A. pedunculatum presented two whole CMA+ chromosomes, a third band corresponding to secondary constriction, also representing an heteromorphic pair of chromosomes, and no DAPI+ bands. A. peregrinum

presented seven CMA+ bands, two interstitial, three telomeric and two corresponding to secondary constrictions. A. peregrinum was the only species that presented DAPI+ bands (9): eight telomeric and one interstitial. The difference observed in CMA and DAPI banding patterns reinforces the taxonomic distinction and indicates that probably there is no gene flow between species, at least between the populations analyzed. The presence of what is apparently a whole CMA+ chromosome in A. pedunculata could indicate the presence of a B-chromosome, but up to date there’s no record of B-chromosomes for the genus. Despite being collected from different States, A. peregrinum and A. campicola are found as synchronopatric in one population from Uberlandia, where artificial hybridizations have been performed and there is a chance that they might naturally interbreed. Further cytogenetic studies will be performed, like C-banding and FISH (45S rDNA), and karyotypes compared with individuals from the synchronopatric
FAPEMIG/CAPES/CNPq.
population.
P1100 – ePoster
Floristic analysis of the Indian vegetation
Gandhi, KN 1 , Shrestha, UB 2 , Ramamurthy, KB 2
1
2
Harvard University Herbaria, Cambridge, USA;
University of Massachusetts, Dept of Biology Boston,
USA; 3 Ashoka Trust for Research In Ecology and
Environment, Bangalore, India
Flora of India is a collaborative project between the
Ashoka Trust for Research in Ecology and Environment
(Bangalore, India), the University of Agricultural Science
(Bangalore), the Missouri Botanical Garden (St Louis,
Missouri), the Harvard University Herbaria (Cambridge,
Massachusetts), and the Royal Botanic Gardens, Kew
(UK). The uniqueness of this floristic project is to provide type information for most names (accepted & synonyms) and also GIS maps and images. The APG-III scheme of classification is followed. Joseph Dalton
Hooker’s pioneer works, viz., Flora Indica (1855; coauthored by T. Thomson), Flora of British India
(1872–1897; coauthored by many botanists), and A
Sketch of the Flora of British India (1904), have been the basis for all subsequent floristic, ecologic, and phytogeographic studies within India. The country, with an area of 1.26 million sq. miles and a coastline of 3,500 miles, has four biodiversity hotspots: 1) Himalaya, 2)
Indo-Burma, 3) Western Ghats (all on the mainland), and
4) Nicobar Islands (part of Sundaland hotspot). The
Indian floristic region, which belongs to the Paleotropical
Kingdom, is estimated to have 18,000 vascular plant species. The class Magnoliopsida has ca. 175 families, ca. 1,950 genera, and ca. 13,500 species (ca. 4,000 endemic), and the class Liliopsida has ca. 40 families, ca.
675 genera, and ca. 4,200 species (ca. 1,100 endemic).
The dominant families include: 1) Orchidaceae , 2)
Fabaceae (Leguminosae), 3) Poaceae, 4) Rubiaceae, 5)
Euphorbiaceae, 6) Acanthaceae, 7) Asteraceae, 8)
Cyperaceae, 9) Lamiaceae, and 10) Urticaceae. The dominant genera (>100 species) are: 1) Bulbophyllum , 2)
Carex , 3) Dendrobium , 4) Erica , 5) Eugenia , 6) Ficus , 7)
Habenaria , 8) Impatiens , 9) Pedicularis , and 10)
Strobilanthes s.l
. Unique distributions include: a) in
South India: Acanthaceae, Lamiaceae, Podostemaceae,
Rubiaceae, Ceropegia, Crotalaria and Tephrosia; b) in
South India and Himalaya: Annonaceae and Impatiens; c) in South India and Northwest India: Euphorbiaceae; d)
Gangetic Plains (lower): Nepenthes; e) Himalaya:
Brassicaceae and Caryophyllaceae (both Mediterranean families), Fumariaceae and Papaveraceae (both North
Asian families), Primulaceae, Ranunculaceae, Rosaceae,
Saxifragaceae, Rhododendron (forms a belt at the base of the mountains), and Vaccinium. The Dipterocarpaceae and Pentaphyllaceae link India with Malaysia, and
Capparaceae (Cappareae) and Ancistrocladaceae suggest an influx from Africa. The Tibetan Plateau seems to act as a phytogeographic barrier between southern China and
India. The genera Castanea, Fagus , and Tilia , which have temperate distributions, are absent in temperate
Himalaya. The Gondwana fossil flora includes
Glossopteris and Vertebraria .
P1079 – ePoster
1
Balancing plants and water use: a classical perspective
Balbach, M 1 , Balbach, H 2
Illinois State University, USA; 2 USA ERDC,
Champaign, IL, USA
When did humans merge the needs of crop and ornamental plants with water management? The answer appears to be ‘Almost as long as agriculture has been practiced in arid lands.’ The history of water management for agriculture in Southwest Asia and North
Africa is long and well documented, but we are now aware of irrigation works in Meso and South America which surely were independently developed. Beyond pure agriculture, one of the most culturally interesting applications may be the association of water and plants in what we term the classic Persian garden. Many of us have read, or heard in a lecture, that the view of a garden as an earthly paradise may be attributed to the Islamic religious culture. To what degree is this a tenable explanation? Or, is there any truth at all to it? Is it all myth? Western history does show us that the walled garden, with a central water feature, has been repeated for a millennium throughout the Middle East and North
Africa, in Spain, and even in India. In all these locations, the garden design has been associated with Islamic rulers and culture. But if it did not come from Islam, then where did this concept originate? It is reasonable to believe that beyond pure agriculture, cultures based in arid lands would appreciate the visual comfort of running water and lush, green plantings. In contrast to the harsh landscape outside the walls, these gardens do provide a quiet, tranquil vision…in some ways, an escape from the desert reality. And, since the Middle Ages, western culture has more or less equated the cultures of the arid
Middle East and southwest Asia with Islam. But it was not always so. Islam as a political and cultural force is a relative newcomer to the region. By the 5th and 6th centuries BCE, a millennium before Islam’s rise across the region, the great Persian rulers Xerxes, Darius, and
679

Alexander had spread their culture from the Bosphorus to the Indus Valley. It was this culture throughout the
‘whole known world’ that actually promoted the vision of a walled garden rich with plantings and water features.
So, while almost all of our existing examples of the gardens representing paradise date from the reigns of
Islamic rulers in the 10th through 15th centuries, these are just the most recent and best preserved examples of a tradition more than 2,000 years old. And, so far as we may determine, there is really no special religious significance to the gardens…the ‘paradise on earth’ concept may well be just a myth…wishful thinking. It has, however, left us with many beautiful examples of a traditional marriage of plants and water which may be enjoyed by us in the 21st century.
P1080 – ePoster
Indigenous knowledge, uses, and management of
Marula (
Sclerocarya birrea
Hochst.) in Limpopo
Province, South Africa
Bhat, RB 1 , Mabala, G 1 , Tshisikhawe, P 1
1 Dept of Botany, University of Venda, Thohoyandou,
Limpopo Province, South Africa
Sclerocarya birrea or Marula tree is indigenous to the miombo woodlands of southern Africa and the Sudano-
Sahelian range of West Africa and considered as one of
Africa's botanical treasures. It belongs to the mango family (Anacardiaceae). Traditionally the fruits and nuts are used as food, and bark and leaves are used for curing various ailments. The tree has got great cultural and social significance. The tree grows from Ethiopia to
South Africa. The primary concern of this investigation is to assess the marginal utilization of this wild tree that is not cultivated for commercial purposes. The present study was undertaken with a view to bridge the indigenous knowledge with the modern utilization of this tree. It is believed with the foreign intervention, urbanization and change of cultural practices; the traditional knowledge of plant uses is slowly being lost from generation to generation. There is an urgent need to record and to make use of the indigenous knowledge for poverty alleviation, job creation and for the welfare of the community before it is completely lost. Indigenous knowledge is equally and well established among the poor and rich in the community. Ripe marula fruits are the important source of food and marula alcoholic and non-alcholic beverages. The fruit quality is affected by sweet taste, size of the fruit and quality and quantity of fruit pulp. Wild marula trees are traditionally spared in the cultivated lands. The marula trees regenerate themselves or by the animals, when the seeds are discarded through the animal dung. The traditional uses of marula trees are widespread and it contributes to combating poverty.
P1081 – ePoster
Plants and society in Guyana: the sexual potency and unknown human well-being cost–benefit dimension in the case of validating a local herbal aphrodisiac
Caesar, JC 1 , Craig, K 1
680
1 University of Guyana, Georgetown
Guyana’s mainstream ethnopharmacopoeia is enriched by a repertoire of forest ethnobotanicals used by local indigenous tribes. While users of herbal remedies attest to their efficacy, there is a paucity of scientific validation. One of the most popular herbal concoctions, especially among men, is the local ‘Kapadulla Tonic.’
This herbal aphrodisiac is derives its name from the vernacular name of the tropical rain forest liana
Tetracera volubilis . The herbal tonic is touted to enhance male sexual performance, increase libido, improve ejaculation and ‘treat’ erectile dysfunction. The tonic comprises eight other plant species to which some molasses and a dash of Guiness stout may be added. We have previously branded the decoction of the Kapadulla alone as Guyana’s ethnoviagra based on preliminary results indicating its ability to increase testosterone levels in male Spragg-Dawley rats (Clay and Caesar 2008).
Literature reports on testosterone levels in birds encouraged us to test the effect of this ethnoviagra on the domestic fowl, Gallus gallus domesticus . Twenty-four newly hatched 3-day-old vaccinated broiler chicks were purchased from the Didco Poultry. Twelve each of the chicks were randomly assigned to control and test groups in separate meshed cages. The control group was fed ad libitum on regular poultry feed and given pure drinking water dispensed from a poultry water canister. The test group was fed poultry feed on which was sprinkled 20 ml of a 10% (w/v) decoction from the chipped woody vine of the Tetracera volubilis (100 g wood chips boiled in
500 ml artesian well drinking water) ad libitum. The test group was given a 10% decoction as drinking water via a poultry water canister. The chicks were cared for over a period of 7 weeks. At the end of the experiment 6 individual chicks each of the control and test groups were sacrificed for blood samples and gross anatomy weight measurements of the unplucked carcass, plucked carcass, liver, heart and kidney. Blood samples were analyzed for lipid profile at the Woodlands Hospital in Georgetown.
We report here on the results of the gross anatomy and lipid profile. Test chicks had significantly larger liver
(ANOVA p = 0.00001); larger kidneys (p = 0.0003); and hearts (p = 0.0002). Plucked weights were slightly greater in test plants but not statistically significant . Total cholesterol in test chicks decreased by 11% but this was not statistically significant. Triglycerides in test chicks were significantly decreased by 52% compared with control chicks (p = 0.02). LDL increased in test chicks by
35%, but was not statistically significant, while HDL decreased by 10% in test chicks though not statistically significant. We believe the use of this plant-derived aphrodisiac may have some effect on lipid profiles and this may impact cardiovascular health in the chicks with analogous potential health risks needful of careful examination. An extended study is planned.
P1082 – ePoster
Application of local trees in landscaping a university garden
Chen, Q 1 , Chen, GL 1
1 Inner Mongolia University, Huhhot, PR. China

The university garden is an organic combination of natural landscape and sense of culture. A high diversity of plant community promotes the ecological benefit of the university garden, and it is an essential component of content of university garden. Various gardening styles enrich university’s cultural diversity, meeting students’ aesthetic demands. Application of local trees increases the diversity of landscaping trees, facilitating the expressions of university spirit and projecting local culture. Stressing and appropriately applying local trees in landscaping university garden provides an efficient way to beautify the university campus. We have reported that while 85 species of woody plants are applied in the three major universities in Hohhot, Inner Mongolia, which belongs to 29 families, only 18 species in 12 families are local trees to Hohhot. We also reported that
21 species in 9 families are found in Xiaojinggou reserve in northern Hohhot. While rarely seen in the university campus, they grow well and are highly resistant to adverse conditions, bringing ecological and educational benefits to the teachers and students. Local trees should play a major role in university gardening.
P1084 – ePoster
Flax (
Linum usitatissimum
): an ethnobotanical study of the abandoned fibre and oil crop in Turkey
Uurlu, Z 1 , Dönmez, AA 1
1 Hacettepe University, Turkey
Based on the archaeobotanical evidence from southeastern Turkey and other countries in the Fertile
Crescent, flax was used by local people at least 10.000 years ago. The earliest finds of flax remains in Turkey come from a Neolithic settlement, Çayönü, in the southeastern part of the country. Flax was one of the important crops for fibre and oil production during the
Ottoman Empire time . It was widely cultivated throughout the Black Sea region and inner parts of the area until 40 years ago. Due to changing agriculture policy and lifestyle of the local people, it has been abandoned. Beside this, many of the local organizations, both governmental and non-governmental, have been trying to keep this agricultural heritage by various activities. During a floristic work in Sinop in northern
Turkey, where flax agriculture had been extensively done in the past, it has been noticed that one of the villages keeps the flax tradition alive. A case study has been carried out in this village and various hand crafts have been observed there. Various stages of flax agriculture tradition including harvesting, obtaining the fibres, manufacturing the linen and producing various clothes and other handmade instruments have been documented and an overview of flax culture in Turkey have been discussed from different points of view.
P1085 – ePoster
Considering biological and cultural diversity in the context of botanical conservation strategies
Dunn, C 1
1 Lyon Arboretum, USA
Concerns continue to mount regarding potential impacts of global climate change, habitat loss, and other environmental changes on the world’s flora . Over the millennia, peoples and cultures across the world have developed deep and unique relationships to the land and have discovered many valuable uses on plants in their areas . Just as floristic diversity is eroding, so too are cultural and linguistic diversity . UNESCO estimates that
50% of all languages are endangered, with one language being lost every 2 weeks . As plants become increasingly rare, and ultimately extinct, the cultural values and significance they represent disappear . Thus, cultures risk losing critical elements of their identity . By the same token, as cultures and languages are lost, we lose traditional knowledge about plants, unique perceptions of time and of nature, and intellectual wisdom .
Consequently, botanic gardens and other conservation organizations should consider both biological and cultural diversity within their conservation programs . It is not enough to consider just the effects of environmental change on plant life within the current context of the GSPC and CBD (particularly Article 8(j)) .
Rather, we must actively engage in understanding the broader impacts of environmental change to ‘biocultural’ diversity.
P1088 – ePoster
Temporal variation in wild plant pathosystem
Lactuca serriola
-
Bremia lactucae
Lebeda, A 1 , Petrzelova, I 1
1 Palacky University In Olomouc, Faculty of Science,
Dept of Botany, Czech Republic
Research on spatial and temporal variation in the wildplant pathosystem, Lactuca serriola (prickly lettuce) –
Bremia lactucae (lettuce downy mildew), has been carried out in the Czech Republic in 1998–2009.
Altogether 488 isolates of B. lactucae were examined for the presence of 32 virulence factors (v-factors). Over one hundred different virulence phenotypes (v-phenotypes) were recorded; however, virulence structure changed substantially, each year only one or a few v-phenotypes were recorded as dominant. A low number of vphenotypes was recorded during the whole period
(Lebeda and Petrželová 2004), comparing to extremely high uniqueness of v-phenotypes found on L. sativa at the same time (Lebeda et al. 2008). Most common vfactors in B. lactucae populations match some of the race-specific resistance genes/factors (Dm genes or Rfactors) originating from L. serriola . The highest frequency was recorded by some v-factors (e.g. v7, v11, v15–17, and v24–30). In contrast, v-factors (v1–4, v6, v10, v12, and v13) matching Dm genes located in L. sativa were very rare. This demonstrates the close adaptation of B. lactucae virulence to the host ( L. serriola ) genetic background (Lebeda et al. 2008). For many v-factors (v11, v14, v16, and v25–28), fluctuations were observed, some (v14 and v17) shifting to higher frequencies, and vice versa (v5/8 and v23). Surprising was occurrence of v-factors matching quite new resistances (R35, R36 and R38) derived from wild
Lactuca spp. ( L. virosa, L. saligna and L. serriola , resp.), which have been recorded in populations of B. lactucae sparsely since 2004, virulence to R38 increased markedly
681

in 2008. Increasing frequencies of v-36 and v-38 has been previously recorded for B. lactucae originating from
L. sativa (Lebeda et al., 2008). In four European countries (Czech Republic, Germany, Netherlands and
UK) variation in resistance of host ( L. serriola ) populations (Lebeda et al., 2007) to B. lactucae was studied. Generally, European populations of L. serriola have been shown to be highly susceptible to B. lactucae .
However, large variation in L. serriola resistance was found both among and within populations of individual countries. There was a clear gradient of increasing uniformity of race-specificity, moving from Central to
Western Europe and United Kingdom, as well as a decrease in the diversity of R-phenotypes (Petrželová and
Lebeda, 2010) . This research was supported by grant
MSM 6198959215 .
P1089 – ePoster
Ferns in art: the last 150 years
Palacios-Rios, Monica 1
1 Instituto de Ecología, AC, Mexico
During the Nineteenth and early Twentieth centuries, there was a phenomenon among 'Victorians' known as the fern craze, due to their special predilection for ferns.
This passion was expressed in the collection of ferns, taking new or rare species to the United Kingdom (in particular) and, specially, in the elaboration of a wide array of objects (either decorative or for daily use), in which the graceful and pleasant shape of ferns was used.
Of the utmost importance for this fancy was the observations done on the Maori culture of New Zealand, where ferns are a center-piece of the culture. The beauty of the Maori culture will not be addressed here, but only its great influence of fern shape and form on this era of the United Kingdom. Fern designs were present on objects of the widest kind. For instance, in ceramics, porcelain, glass (crystal), different metals, textiles, wood, printed stationary, rocks, candles, curtains, flowers pots, chimes, stamps, and many other kinds of materials. A great many of these objects were manufactured by
Darwin Country, who fabricated iron objects for the
Coalbrookdale Company, and ceramics of well-known brands such as Wedgwood, Dudson, Staffordshire, and
Shropshire. The use of the Wardian case, as useful since that time, to maintain the ferns out of the air pollution, bad weather. Here I present a portion of this well-rooted, interesting and dominant fashion within the English society. This great influence also permeated other societies where this phenomenon is usually addressed as in past tense, but which still prevails among people of different countries and regions of the world. I have made a compilation of the records of this vast array of art and every-day objects with designs that include the fresh, graceful, pleasant and elegant shape of fern fronds and the fiddleheads (prefoliaciones, báculos, colas de violín, colas de gato, colas de mono, camarones, korus, cayados, etc.), which I want to exhibit in a grouped, systematized and orderly way, as part of ©Proyecto Helechos – Ferns
Project (http://www.helechos.com.mx). This topic should be addressed in contemporary art, which is still relevant and vibrant in the Twenty-first century. This information is part of a future book that I am preparing on the topic.
682
P1092 – ePoster
Diversity and seasonal occurrence of fungi from some
Indian soils
Rane, G 1
1 Moolji Jaitha College, India
Studies on soil fungi from Khandesh region of
Maharashtra was carried out for 3 years. The three districts from Khandesh i.e. Jalgaon, Dhule and
Nandurbar were selected for soil sample collections. In each district 8 sites were identified. The 8 sites included
2- cultivated soils, 2- river soils, 2- uncultivated soils and
2- forest soils. Thus 24 sites were selected for soil sample collection from 3 districts. Soil samples were collected in every season, i.e. winter, summer and rainy season for three years. All these soil samples were brought to the laboratory and inoculated on different culture media.
Observations were made. Identification was done along with camera lucida and photomicrographs. A total 261 species belonging to 57 genera are isolated from the soil of Jalgaon, Dhule and Nandurbar districts of Khandesh.
Deuteromycetes members are dominant in all the soils with Aspergillus as a dominant genus. From the species isolated, 1 genus and 1 species of Dyctyostelium from myxomycetes, 2 genera and 2 species from
Mastigomycotina, 9 genera and 32 species from
Zygomycotina from which commonly occurring are
Mucor and Rhizopus , 4 genera and 28 species from
Ascomycotina with dominance of Chaetomium . 41 genera and 193 species from Deuteromycotina with
Aspergillus as dominant genus. 5 different Mycelia sterilia are also isolated from various soil types. Among the soils of three districts, 50 genera and 138 species are isolated from Jalgaon district, 41 genera and 182 species from Dhule district and 35 genera and 171 species from
Nandurbar district are isolated. So, highest numbers of species are isolated from various soils of Dhule district.
All these fungi isolated showed remarkable seasonal variation. Some of the species are season specific while many are occurred during all seasons. From all the species isolated, maximum number of species are isolated from winter season followed by rainy season and minimum number of species are isolated from summer season. In the soils of Jalgaon district maximum number of species, i.e. 95, are isolated during winter season while during rainy season the numbers of species isolated are
85. Minimum numbers of species are isolated during summer season, i.e. 73 species. In Dhule district maximum number of species, i.e. 110, are during summer followed by 100 species during rainy season. Minimum numbers of species are isolated during winter season, i.e.
93 species. Nandurbar district also showed seasonal variation maximum number, i.e.108 species are isolated during summer followed by 106 species from rainy season. Minimum number of species are isolated during winter season, i.e. 97 species. Present study gives the complete taxonomical description of the fungal species, its camera lucida sketch and photo micrograph which made the identification easy and accurate. This work will be a good piece of work in fungal taxonomy. There are new reports of different soil fungi. 5 species are recorded as new species. 167 species are reported as new to
Maharashtra, 56 species as new to India. 201 species as

new to soils of Maharashtra and 100 species as new to soils of India.
P1094 – ePoster
Use of city parks in the teaching of botany
Vieira, EDR 1 , Schwarz, EDA 2
1 Universidade Federal Do Paraná, Brazil
This paper aims to assess the use of city parks as a methodology for teaching botany. The study, in which eighth grade students from a public school in the state of
Parana analyzed the local flora, was conducted in St
Lawrence Park (Parque São Lourenço), located in
Curitiba, Parana, Brazil. The project was developed through Paraná State Government's Educational
Development Program (PDE), by the Public Education
Network of the State of Paraná, through which a certain number of teachers is selected annually to study and develop, through scientific papers, methodologies that can enhance teaching in state schools. The study showed that the students were able to recognize all of the botanical groups (algae, lichens, bryophytes, ferns, gymnosperms and angiosperms, as well as fungi, no longer part of the plantae kingdom), and the students were able to identify eight botanic families among the area's tree vegetation. Thus the project revealed that the park is a highly educational environment, whose potential for teaching should be explored pedagogically.
With field trips, the science classes were very dynamic and as the students conducted basic scientific research, they were able to become completely involved in the learning process. Throughout the experiment, the students socialized and cooperated during both computer science and field classes: they not only filled each other in on what was being discussed, and which procedures were to be adopted, but also encouraged each other's participation. The experiment resulted in the development of a CD-Rom about the history of the park and the Belem River, containing pictures taken in the park, a description of the vegetation from the area, and collections of dried specimens, fruits and seeds. This set of materials was developed by the students under the guidance of the science teacher .
P1095 – ePoster
Communities: plants and people
1
Sutcliffe, J 1
Manhood Wildlife and Heritage Group, UK
Introduction: the Convention on Biological Diversity
(1992) stresses the need for local communities to be involved in protecting their local wildlife . In the UK, an innovative project was established by local people in
Selsey, West Sussex . As a contribution to the
Millennium, a group embarked on mapping the vegetation of the parish and they also produced a map of the village . Method: The Selsey Parish Mapping Group was launched at a meeting in March 1998 and volunteers put down their names to help. Contact was made with the local landowners, interested groups and the Parish
Council. A grant was obtained from Rural Action and
Sussex Wildlife Trust provided training in vegetation mapping. The group went out on an exercise together to ensure that they would map areas in the same way. The
Parish was then divided into four and mapped by systematically walking the fields . The results were put onto one large map and colour coded as required . This baseline map now forms a tool used by the group to protect remaining areas of habitat and species interest in dialogue with local people and specialist interest groups .
Rare plant records are provided by the local Sussex Rare
Plants Group and the Biodiversity Records Centre supplies maps of the area incorporating the records for rare species . The Chichester Biodiversity Plan has used the project as a model to encourage other communities to adopt the idea . Vegetated shingle: the south coast of
England has 25% of the world resource . Selsey Parish contains some degraded examples and this damage is being addressed . The local school has built a demonstration shingle garden in their grounds . Results: the group produced the first UK Parish Biodiversity
Action Plan and works with all key local partners to put it into effect – West Sussex County Council, Chichester
District Council, Pagham Harbour Local Nature Reserve,
Sussex Wildlife Trust, the town Council and local landowners . They have ensured that the targets in national, regional and local plans are taken into account and began work by strengthening local hedgerows and identifying areas suitable for additional tree planting . The
Parish lies between a geological SSSI at West Wittering,
Chichester Harbour SSSI and Pagham Harbour Local
Nature Reserve so the main contribution is tackling issues in the wider countryside. The group has extended to cover the 11 parishes on the Manhood (derived from
Main wood) Peninsula and is engaged in a project to identify ecological networks and carry out conservation projects in key areas . An important aspect has been the surveys of Water Vole, a BAP species . The full time staff member, Sarah Hughes, is now funded by a planning mitigation S106 agreement for the next five years.
P1096 – ePoster
An evaluation of threat of bark harvesting in Venda region, Limpopo Province, South Africa
Tshisikhawe, MP 1,2 , van Rooyen, MW 2 , Bhat, RB 1
1 University of Venda, South Africa; 2 University of
Pretoria, South Africa
Ethnobotanical studies indicate that the growing trade in indigenous medicinal plants in South Africa is posing a threat to the conservation and preservation of many plant species. Trade in medicinal plants has become a way of making a living for some people and pharmaceutical companies. The medicinal flora of Venda region consists of a variety of species, which potentially may provide therapeutic agents to treat different diseases . This study is an investigation into the pattern of trade in medicinal plants by local markets in Venda region, Limpopo
Province, South Africa . Indications are that bark harvesting may threaten some of the plant species . Fifty eight medicinal plant species are harvested for their medicinal barks . The study also revealed important medicinal plant species within the trade as well as the impact of middlemen in trade of medicinal plants.
683

P1097 – ePoster
Changes in fungal diversity of indoor air of the M.S.
University of Baroda, India and its association with surrounding plants
Vyas, N 1 , Soni, S 1 , Arya, A 1 , Rajeshirke, S 1
1 MS University of Baroda, India
Fungal spores are among the most commonly encountered bioparticles in atmosphere. The composition of air borne fungal spores is characteristics of each biogeographical zone depending upon the type of vegetation, their number and type depends on the time, day, weather and season. The study of fungal spores is of immense importance due to its role in the prediction of plant and human diseases and also due to microbial deterioration of the materials like paper, textiles, printed surfaces etc. The importance of such studies, can be helpful to phytopathologists and doctors to prevent various diseases of plants, animals and human beings.
The M.S. University of Baroda is a place spreading in 18 km 2 where more than 35,000 students get educated in 13 different faculties . An effort was made to find out the indoor aero-mycoflora in 7 different faculties with 2 different locations by gravity fall method. Three different petri dishes of 100 mm dia. were exposed for
10 min . Data were recorded in different months of a year.
The maximum number of fungal colonies were recorded in the Faculty of Arts in the French department and the minimum number of colonies were found in the Faculty of Science in the department of Environment Science.
Many of the fungal species found in petri dishes belonged to the genera Aspergillus, Alternaria,
Cladosporium, Penicillium, Rhizopus , and Trichoderma , etc. The fungi like Alternaria and Cladosporium were also reported from the trees growing in the campus.
684

Abeli, Thomas
Aberlenc, Frédérique
Abiko, Tomomi
Abou-El-Seoud, Dalya
Acevedo, Pedro
Ackerman, J.D.
Adam, Iris
Adam, Zach
Adamek, Markus
Adekanmbi, Olusola
Ades, Peter K
Adeyemi, Temitope
Adler, Sophie
Adýgüzel, Nezaket
Afshar, H
Aghaalikhani, Majid
Agosti, Donat
Aguilera, Justina
Aguirre-Hernández,
Rebeca
Agulló, J.C.
Ahamadeen Nagoor,
Mohamad Mubarak
Ahamed Abd El-D,
Mohamed Salama
Ahmad, Altaf
Ahmadi, Shahla
Ahmadian, Najmeh
PL = Plenary talk; KNS = Keynote Symposium talk; PU = Public Talk; SYM = General Symposium talk; P = ePoster or Poster
Author Reference Author Reference
A. Dönmez, Ali P1033 Ahmadiani, Sara P0462
A.C. Dos Santos,
Deborah Yara
Aaslo, Per
Abbaspour, Hossein
Abbaspour, Nasser
Abbott, Richard
Abd El Wahed,
Mohamed,
Abdel-Hady, Mohamed
Sarwat
Abdolmaleki, Parviz
Abdul Rahman, Amal
Abe, Katsumi
Abe, T
P0297
SYM032
P0458, P0459,
P0460, P0549, P0550
P0237
SYM157
SYM031
P0238
P0487
P0500
P0277
P0085, P0239,
P0272, P0283,
P0303, P0633
P0002
SYM162
P0455
P0125
P0708
SYM142
SYM124
SYM068
P0102
P0146
SYM121
P0912
SYM162
P0728
P0460
P0517
SYM136, P0712
SYM119
SYM114
P0834
SYM028
SYM031
SYM030, P0563
SYM009
P0284
Ah-Peng, Claudine
Ahuja, P S
Ahvazi, Maryam
Aiba, Shin-Ichiro
Ainouche, Abdelkader
Ainouche, K
Ainouche, M
Ainouche, Malika
SYM158
SYM159, P0511
SYM177
SYM110
P0976
SYM157
SYM157
P0976
Ainsworth, Elizabeth A. SYM156
Aizawa, Katsunori SYM026
Aizen, Marcelo
Akashi, Tomoyoshi
SYM102B
SYM145
Akbas, Halide
Akiyama, Kohki
Akuzawa, Sayuri
Al Dhaheri, Shaikha
P0351
P0558
P0277
P0169
Al Mehairby, Mohamed P0169
Al Shehbaz, Ihsan SYM097
Alagumuthu, G
Alamsyah, Firman
P0322
P0714
Albertini, Emidio
Albion, Rebecca L
Albrecht, D.E.
Albrecht, Veronica
SYM162
SYM045, P0373
SYM084
SYM068
Albuquerque, Elaine
Albury, Mary
Aldasoro, Juan Jose
Alex, Morno
Alexander, Patrick
Alexandre, Fournier-
Level
Alfayate Casañas,
Carmen
Ali, Haidar
Allaby, Robin
P0715
SYM156
SYM103
SYM008
P0716
SYM069A
P0609, P0610, P0611
Allen, Chris
Allen, R.S.
Allison, Allen
Allkin, R
Allkin, Robert
Allphin, Loreen
P0221
SYM013
SYM020
SYM060A
SYM166, P0078
KNS10, SYM118
KNS10
P0716
685

Author
Almeda, Frank
Almeida, Ana Maria
Alonso, M.A.
Alonso-Peral, M.M
Al-Shehbaz, Ihsan
Altioglu, Yusuf
Altman, Naomi S.
Reference
P0895
SYM069
P0834
SYM060A
SYM097, P0716
P0005
P0386
Altulu, Yusuf P0005
Álvarez-Dávila, Esteban SYM164
Alves, Marccus
Alzate-Marin, A.L.
P0618
P0065
Amaral, Maria
Ames, Mercedes
Amien, Suseno
Amirahmadi, Atefeh
P0717, P0718, P0998
SYM117
KNS04
SYM130
Amity, Wilczek
Amos, Nevil
Amthor, Jeffery
An, M.
Anamthawat-Jónsson,
Kesara
Anderson, Cajsa Lisa
Anderson, Charles
Anderson, Gregory J
Anderson, Ian
Anderson, Peter
SYM069A
SYM078
KNS06
P0532
P0240, P0719,
P0720, P0768
SYM103, P0721
PL10, SYM059
P0951
SYM016, P0223
SYM055
Anderssen, Bob
Andhale, Balasaheb S.
Andrade, M.J.G.
Andrew Leitch, Andrew
R.
Andrew, Rose
SYM060B
P0317
P0802
P0845
KNS02, SYM120
Andriansolo, Domohina SYM066
Angaji, S. Abdolhamid P0799
Ángeles, J.E
Angeluzi Jardim,
Veridiana
Anne-Caroline,
Cosendai
Ansari, Ziya
SYM011
P0138
P0074, P0824
Antonelli, Alexandre
Antonio, Baltazar
Aoki, Toshio
Aoyama, T.
Apel, Klaus
Apparao, Boddu
Appelhans, Marc
P0908
SYM077B, SYM103
P0361, P0451, P0455
SYM145
P0559
SYM068
P0241, P0464
SYM115
686
Author
Appels, Rudi
Applequist, Wendy
Reference
SYM048, SYM159,
SYM160, SYM161,
SYM162
P0722, P0814
SYM009 Araghi, Mohamad
Kazem
Arakawa, O.
Araki, T
P0426
SYM042
Aranda-Rickert, Adriana P0007
Araujo, Ana Claudia SYM091
Araujo, Dorothy
Arbor, Ann
Arcela, Vicente
Archibald, Sally
P0118
SYM004
P0008
SYM014
Arda, Hayati
Ardi, Wisnu H.
Arenque, Bruna
Argall, M
Arias, Nadia S.
Arikawa, Tomotsugu
Arkhipova, Tatiana
Armstrong, Doug
P0356
P1074
P0009
P0164
P0588
P1056
SYM036
KNS08
Arrighi, Jean-François
Arroyo, Mary
Arya, Arun
Arya, Ramdas
Arya, Sarita
Asadulaev, Zagirbeg
Ascensão, Lia
Asgharzadeh, Ahmad
Ash, Julian
Ashman, T.-L.
Ashton, Paul
Asmussen-Lange,
Conny B.
Aspinall, Sue
Assadi, Mostafa
SYM052
SYM014, SYM167
P0010, P1097
SYM163, P0778
P0252
P0902
SYM142, P0648
P0286
SYM126
SYM038B
SYM091
SYM109
Asthana, A.K.
Åström, Helena
Atalay, Z.
Atawongsa, K.
Atherton, Robin
Atique, Gabriela
Atkins, Craig A
Attia, Kotb
Atwell, Brian
Aubriot, Xavier
Auer, Paul
SYM063
P0799, P0843
SYM087
SYM022
P0770
SYM171
P0420
P0012
SYM052
P0438
SYM012
P0814, P1040, P1041
KNS01

Author
Austerlitz, Frederic
Austin, A.D.
Auvray, Gaëlle
Awano, Tatsuya
Ayabe, Shin-Ichi
Ayers, Tina
Aygoren, Deniz
Ayre, David J.
Azevedo, Aristéa
Azevedo, Patrícia
Helena
Babourina, Olga
Bachelier, Julien
Bacic, Anthony
Backlund, Anders
Backlund, Maria
Bacon, Christine
Badea, Monica
Luminita
Badger, Murray
Bækgaard, Lone
Baêta, Hudson
Baeza, Carlos
Bagler, G.
Bagniewska-Zadworna,
A
Bahk, Jeong Dong
Bai, Mingming
Bai, S-N
Bailey, C. Donovan
Bailey, Joseph
Bailey, R.I.
Bailey, Tanya
Bainard, J.
Bainbridge, Susan J
Bajwa, Rukhsana
Baker, William J.
Bakke Westergaard, K
Balao, F.
Balbach, Harold
Balbach, Margaret
Baldauf, Sandra
Baldwin, Bruce
Ball, Marilyn
Ballard, Harvey
Ballhorn, Daniel J.
Ballizany, Wouter
Reference
SYM015
SYM169
SYM129
P0699
SYM145
SYM172
SYM105B
SYM127
P0117
P0367
P0231
SYM080
SYM046, SYM059,
P0589
SYM145
SYM145
SYM144B
P0585
SYM068
P0231
P0196
P0067
P0511
P0437
P0684
P0815
P0870
P0716
SYM149
P0072
P0036, P0090
SYM087
P0015
P0304
KNS09, SYM109,
SYM144B, P1015
SYM085
SYM126
P0014, P1079
P1079
P1013
SYM120, P0015
SYM022, SYM049
SYM172
SYM035
P0244
Bargel, Hendrik
Barker, A
Barker, Gary
Barker, Michael
Barker, N P
Barker, W.R.
Barkworth, Mary
Barlow, Snow
Barnes, Julia
Barral, E.L.
Barrell, Philippe
Barrett, Brent
Barrett, Craig
Barrett, Luke G
Barrett, Matthew
Author
Balslev, Henrik
Banasiak, Lukasz
Bando, Y
Bang, Sang Woo
Bang, Woo Young
Bani, Barýþ
Banks, Hannah
Bannister, Jennifer
Bannister, Jennifer M
Bannister, Jennifer M.
Bao, Yongde
Barab, Denis
Barbanti, Silvana M.
Barbarà, Thelma
Barbazuk, W.B
Barbedo, Claudio
Barber, Janet C.
Barberena, FFVA
Barblishvili, Tinatin
Barbosa Da Motta,
Lucimar
Barbosa, Luiz C. A.
Barbour, Robert
Barcaccia, Gianni
Barcelo, Juan
Barczi, Jean-François
Bardat, Jacques
Barerro, R
Barfuss, Michael H J
P0661
SYM149
SYM162
SYM024, P0155
SYM067, P0612
SYM158
SYM161
SYM004, SYM094,
P0730, P0947, P1042
SYM151
P1015, P1066
SYM157
SYM038, SYM090,
SYM116, SYM157
SYM129
SYM104A
SYM137
PU01
P0230
P0133
SYM039
P0244
SYM106A, SYM109
SYM078
P0154
Reference
SYM077B, SYM109
SYM079B, P0727
P0679
P0605
P0684
P0728
SYM102A
P0729
P0729
SYM101,
SYM106A, P0729
P0386
P0701
SYM006
SYM002, SYM004
SYM038, SYM157,
P0377
P0467
SYM168, P0779,
P0822
P0018
P0121
P0297
687

Author
Barrett, Russell
Barrett, Spencer. C.H.
Barrett-Lennard, Ed
Reference
SYM091
SYM102B
P0593
Barriel, V.
Barron, Rebecca
SYM116A
SYM056
Barros, Claudia
Barros, Fabio
Barros, T.C.
P0613
SYM105, P0018,
P0926
P0704
Barros-Barreto, Beatriz P0756
Barrow, Russell
Barry, K.M.
Barry, Kerrie
Barthelemy, Daniel
Barthlott, Wilhelm
Bartholomew-Began,
Sharon
Bartish, I.
SYM073, P0920
P0466
PL06
P0687, P0688, P0740
P0095
SYM155
Bartlett, Madelaine
Basnayake, J
Bastian, Meredith
Bastien, Renaud
Batalha, M.A.
Bateman, Ashley
Bateman, Richard
Batstone, Tom
P0072
SYM069
P0290
SYM110
SYM065, SYM067
SYM077
P0961
Baudouin, Luc
Bauer, Brittany
Baulcombe, D.C.
Baumann, Nadine
Baumberger, Rolf
Baumgratz, JFA
Bayle, Vincent
Bayly, Michael
SYM105B
SYM157
P0808
SYM058
SYM060B
SYM064
SYM123
P0018, P0731, P0732
SYM052
SYM095,
SYM104B,
SYM121, P0903
P0287, P0380 Bazanova, Natalia
Bazzolli, Denise
Beard, Katherine F.M.
Beardall, John
Beattie, G. A. C.
Beatty, Mary
Beaulieu, Jeremy
Becerril, Jose Maria
Beck, James
Becker, Matthias
Becquer-Granados,
Eldis
Bedggood, W
P0148
SYM156
SYM026
SYM171
P0589
SYM086
P0060, P0481
P0716
SYM097
P0895
P0164
688
Author
Bedolla-García, Brenda
Y
Beer, A.S.
Beerling, David
Behera, Asmita
Behmanesh, Mehrdad
Bekaert, Michael
Belimov, Andrey
Bell, Charles
Bell, Neil
Bellemain, E
Bellgard, M
Bellot, Sidonie
Benavides, Carlos
Bender, Ricci
Benjamin, Smith
Benko-Iseppon, Ana
Maria
Benlloch-Gonzalez,
Maria
Bennett, Jeff
Reference
P0733
SYM090
SYM040, SYM051
P0296
P0488, P0543
SYM157
SYM036
SYM086
SYM122
SYM013, SYM088
SYM160, SYM161
SYM157
P0407
SYM058
SYM026
SYM004, SYM077B
SYM034
Bennett, Thomas
Bennetzen, Jeff
Benson, Doug
Berding, N
Berg, C.V.D.
Berger, Brent A
Berger, Frederic
Berger, Madeleine R.
Berges, H.
Bergstrom, Dana
SYM005
SYM041
SYM161
SYM133
P0290
P0802
SYM108
KNS01, SYM062
P0845
P0429
SYM022
Berkowitz, Oliver SYM052, P0447,
P0591
Bermingham, Eldredge P0910
SYM106B Bernardes-De-Oliviera,
M.A
Bernardes-De-Oliviera,
M.E.R
Bernardini, Benedetta
Bernhardt, Karl-Georg
SYM006
P0737
SYM082
Bernhardt, Peter
Bernoux, Maud
Berr, Alexandre
Berry, Joseph
Berry, Paul
Berveiller, Daniel
Bessega, Cecilia
Bester, Stoffel Petrus
Betsuyaku, Shigeyuki
Beule, Thierry
PU03, SYM142
SYM055
SYM123
SYM068, SYM156
SYM004, SYM081
SYM105A
P0381
P0738
P0442
SYM162

Author
Beveridge, Christine
Bezerra, E.L.S.
Bhalla, Prem L
Bhardwaj, P.K.
Bhat, R.B.
Bhushan, Shashi
Bickford, Chris
Bidartondo, Martin
Biere, Arjen
Biffin, Ed
Binney, Allan
Biradar, Sanjay
Birch, Joanne
Birch, Paul
Bisby, Frank
Bitonti, Beatrice
Bittrich, Volker
Bjarnholt, Nanna
Bjora, Charlotte
Black, M
Blackman, Chris
Blanchette, Mathieu
Blanchoin, Laurent
Blanchon, Daniel
Blanco, José Luis
Blank, Stephan
Blattmann, Peter
Blattner, F. R.
Blomstedt, Cecilia
Bloomer, Rebecca
Boatwright, James
Stephen
Bobrov, Alexey
Bocayuva, Melissa
Boddu, Jeevakoti
Apparao
Boedeker, Christian
Boessenkool, S
Bogdanoviæ, Sandro
Bogdanowicz,
Agnieszka M.
Bogler, David
Bogner, J
Bohman, Bjorn
Bohnert, Hans
P0672
P0020, P0148
P0241, P0464
SYM075
SYM088
P0030
P0021
P0199
SYM106B
P0921
SYM041
SYM071
P0717, P0718, P0998
SYM053
SYM117A
SYM161
SYM056
SYM096
SYM061
SYM018
P1052
SYM004
P0470
P0928
SYM024, SYM053,
SYM054 P0263,
P0382, P0465
P0362
SYM089, SYM129
Reference
SYM153, P0592,
P0594
P0107
SYM060
P0511
P0061, P0099,
P1080, P1096
SYM159
SYM097
SYM040, SYM087
SYM036
SYM007,
SYM049B,
SYM115, SYM119,
SYM124, SYM131
SYM001
P0248
SYM108, SYM117B
KNS08
KNS08
Author
Bohs, Lynn
Bolinder, Kristina
Bona, Cleusa
Reference
SYM107, P0791
P0222
P0658
Bond, Charles SYM055
Bond, William KNS05, SYM014,
P0099
Bonifacino, J. Mauricio SYM120
Bonnett, Graham P0312
Boonpeng, Chaiwat
Boonpragob, Kansri
Boonprasert, R.
Boonsirichai,
Kanokporn
Borchsenius, Finn
Borem, Aluizio
Borgeault, Maryse
P0022
P0022, P0915
SYM171
P0352, P0353
SYM077B
P0365, P0367
P0176, P0194
SYM096
P0364
Borgen, L.
Borges Do Val,
Aurinete D.
Borja, Guadalupe
Borland, Anne
Born, Céline
Bornman, Janet F.
Borrone, James
Borzak, Christina
SYM069
SYM017, SYM045
P0208
SYM023
SYM069
P0466
SYM070
SYM030
Bosch, Maurice
Bose, Jayakumar
Boss, Paul
Boudaoud, Arezki
Boudko, Katya
Bougoure, J.J
Boujemaa, Nozha
Bourceaux, Angélique
Bourgault, Maryse
Bourne, Aimee
Bowman, John
Boxall, Susanna F
P0448
SYM065,SYM067
P1071
SYM124
P0740
SYM171
SYM034
SYM049B
SYM040, SYM060,
SYM060B
SYM045,P0374,
P0375, P0385
SYM026
P0614
Boyd, Philip
Boyne, Richard
Bozkurt, Meryem
Brackin, Richard
Bradbury, Donna
Braga, Francyane
Tavares
Braga, J. M. A.
Braga, Marcia
Bragg, Jason
P0785
SYM030, SYM031
P0743
P0367
P0715
P0467
SYM037
689

Broennimann, Olivier
Brogliato Junqueira
Engel, Thaíssa
Bromham, Lindell
Bronstein, Judith L.
Brosnan, C
Brown, Adrian
Brown, Austin
Brown, Elizabeth
Brown, Gillian
Brown, Gregory K
Brown, Keri A
Brown, Terry
Brownsey, Patrick
Bruederle, Leo
Brugière, Sabine
Bruhl, Jeremy
Brummer, Tyler
Brummitt, Dick
Brummitt, Richard
Bruneau, Anne
Author
Bramley, H
Brasci Berro, Gabriela
Braukmann, Thomas
Brearley, Francis Q
Breder Eller, Cleiton
Breitwieser, Ilse
Brennan, Adrian
Breuer, J.
Breuer, Klaus
Brewer, Philip B
Brian, Pfleger
Briceno, Veronica
Briggs, B.G.
Briggs, Marie
Bristow, Jim
Broadhurst, L.M.
Broady, P.A.
Brochmann, Christian
Brodersen, Craig
Brodribb, Timothy
Reference
SYM034,
SYM049A, P0307
P0138
SYM124
SYM110
P0023
SYM083, SYM137,
P1008
SYM126, SYM157
SYM034
SYM018
SYM153, P0592
SYM026
P0024
SYM090
SYM128, SYM166
PL06
SYM078, SYM167
SYM026
SYM085, SYM088,
SYM096, P0825,
P0924
SYM049A
SYM014,
SYM049B,
SYM056,
SYM116B, P0355,
P0497
SYM144
P1099
SYM041
SYM150, P0882,
P0883
KNS01, SYM060B
SYM032
SYM088B
SYM104B,
SYM155, P0936
SYM089B
SYM004
SYM013
SYM013
SYM095, SYM133
SYM085, P0744
SYM068
SYM085, SYM091,
P0989
P0014
SYM094
SYM094A
SYM089A
690
Author
Brunken, Ulrike
Brunner, Arco
Brutnell, Thomas P
Bruyns, Peter
Bryant, Gary
Bucci, Sandra J.
Buckeridge, Marcos
Buckley, Yvonne
Budiene, Jurga
Buerki, Sven
Buggs, Richard
Buide, M.L
Buirchell, B.J
Bull Hereñu, Kester
Bull, Roger
Bull-Herenu, Kester
Bumler, Viola
Bundock, Peter
Bunn, Eric
Bunzel, M
Burd, Martin
Bureau, Thomas
Burgman, Mark
Buril, M.T.
Burkhardt, Juergen
Burko, Yogev
Burns, Anna
Burns, Bruce
Burns, Jean H.
Burrows, Geoff
Burton, Rachel
Burzo, Ioan
Bush, Catherine
Bussmann, Rainer
Butcher, Matthew
Butcher, Ryonen
Butler, Don
Buzatto, Cristiano
Byrne, Margaret
Byrne, Mary
Byro-Jilek, M
Byrt, Caitlin
SYM049B
SYM066
SYM001, P0164,
P0263
P0235
SYM014
SYM140
SYM059, P0259,
P0340, P0425,
P0431, P0471,
P0475, P0515, P0589
P0585
P0859
P0097, P0722
P0363
P0026, P0209
P0220
P0975
SYM121, P0026
SYM071
SYM046
SYM028, SYM035,
SYM162
Reference
P0782
SYM039
SYM162
SYM168
SYM022
P0588
P0009
SYM014
P0449
SYM103, SYM115
SYM157
P0754
SYM159
SYM076
P1071, P0938
SYM076A
P0781
SYM088
SYM166, SYM167,
P0084, P0092
SYM046
SYM078, P0025
SYM069A,
SYM096, P0055
KNS07
P0972

Author
Bystriakova, Nadia
Byun, M-O
Cabot, Catalina
Reference
SYM095
P0514
P0155
Cabrera Luna, A.
Caddah, M.
Caesar, John Cartey
P0788
P0998
P0027, P0028,
P0249, P1081
P0702 Caetano, A.P.S.
Cafferty, Steve
Cai, Chao
Cairney, John
Calcoen, Johan
Calió, Maria Fernanda
Call, Christina
Callado, Cátia
Callahan, Damien L.
Callmander, Martin
Calonje, Michael
SYM133
SYM061
SYM016, P0223
SYM045
P0746, P0747
P0716
P0613
P0892
SYM115, SYM118
P0889
Calvillo-Canadell, Laura P0748, P0758
Camargo De Assis,
Marta
Cameron, Duncan
SYM106B
SYM051
Cameron, Kenneth
Campbell, Bruce
Campbell, Christopher
Campbell, Lisa
Cancado, Geraldo
SYM105B, P0793
SYM023
SYM007, SYM119
SYM089B
P0364, P0365,
P0366, P0367
P0040 Cândido, M.V.O.
Cannon, Charles
Cantrill, David
Cao, Daniel
Cao, Jianguo
Cao, Kunfang
Cao, Tong
SYM110
SYM088B, P0887
P0817
P0615
SYM049B, P0588
P0749, P0810, P0869
Caparros Callejo, R. P0051
Cappello Garcia, Silvia P0750
Caraglio, Yves
Cardillo, Marcel
Cardinal-Mcteague,
Warren M.
Cardona-Correa,
Christopher
Cardozo, A. P.
Cargill, Christine
Carling, Jason
Carmello-Guerreiro,
Sandra Maria
Caron, Henri
P0687, P0688
P0933
P0751
SYM040, P0029
P0190
SYM087
P0423
P0616, P0702, P0705
SYM088B
Author
Carr, G.W.
Carter, Clay
Reference
P0903
Carrillo Ruiz, Hortensia P0166
Carroll, Andrew PL10
Carroll, Bernard SYM060A,
SYM060B
SYM058
Carter, Richard
Caruso, Giuseppe
SYM121
SYM011, P0030,
P0250, P0752, P0753
SYM092, P0482 Carvalho, Maria Angela
M.
Carvalho, Monica SYM092, SYM101
Casimiro-Soriguer, Ines P0754
Casimiro-Soriguer,
Ramon
Cassano, Valéria
SYM126, P0755,
P0993
P0756, P0757
Cassman, Kenneth
Castor, C.
Castric, V.
Castro Souza, Vinicius
CASTRO, M.Dem.
Castro, Sílvia
Castro, Víctor
Casu, Rosanne
KNS06
P0389
P0429
SYM131
P0852
SYM105
P0053
SYM160, P0312
Catacora, P
Catalán, Pilar
Catalano, Santiago
Catapano, Terrance
SYM011
P0941
SYM007
SYM136, P0712
Caujapé-Castells, J.
Cavagnaro, Timothy
SYM127
SYM016, SYM035,
P0263
SYM059 Cavalier, David
Cavanagh, Colin
Cavieres, Lohengrin
Celep, F.
Centeno, Danilo
Ceron-Souza, Ivania
Cesoniene, Laima
SYM152
P0218
P0770
P0467
P0910
P0919
Ceusters, Johan SYM045
Cevallos-Ferriz, Sergio P0748, P0758
Chacana, Max
Chacón, Juliana
Chaer, Lia
Chagnoux, S.
Chakraborty, Sukumar
Chakraborty, Tamal
Chamala, Srikar
Chambers, Susan
P0759
SYM106B
P0479, P0480
SYM005
KNS08
P0031
SYM038, SYM157,
P0377
SYM016
691

Chatelain, Cyrille
Chatrath, R
Chatrou, Lars
Chau, Marian
Chauahn, Sudhir
Chaumeil, Philippe
Chautems, Alain
Chavarría Krauser,
Andrés
Chaw, Shu-Miaw
Che, Ping
Cheal, David
Chebli, Youssef
Cheeseman, John
Chelaifa, H
Chen*, G.L.
Chen, Cheng-Wei
Chen, Chuan
Chen, Guilin
Chen, Q.
Chen, Shaolin
Chen, Shilong
Chen, Tao
Chen, Wenyun
Chen, Ying-Ju
Author
Chan, Kai
Chandler, Peter
Chandler, Richard
Chandna, Ruby
Chandore, Arun
Chang, Li-Wan
Chang, Ying
Channing, Alan
Chao, Yi-Shan
Chapman, Eric
Chapman, Sam
Chapman, Scott
Charkhchian,
Mohamad-Mahdi
Charles-Dominique,
Tristan
Chartier, M
Charuvi, Dana
Chase, Mark
Reference
SYM054
SYM055
P0208
SYM030
P0032
P0034
SYM049B,
SYM122, SYM124
P0721
P0760, P0761
P0660
P1066
SYM034, P0439
SYM009, SYM177
P0688
SYM106B
SYM068
SYM057,
SYM105B,
SYM106A,
SYM117A,
SYM123, SYM157,
SYM168, P0850,
P0947, P1042
SYM118
SYM036
SYM104, P1045
SYM166
P0252
SYM088B
SYM077A
SYM065
P0432
P0084
SYM170
SYM065
SYM041
SYM157
P0054
SYM095
P0763
P0368, P0764, P1082
P0368, P0764, P1082
PL10
P0762
P0765
P0033
P0967
692
P0999
SYM094B
SYM077B,
SYM116B
SYM060A
SYM049
SYM036
SYM170
SYM122
P0999
SYM129
SYM095, P0760,
P0863, P0874
P0034, P0766
P0234, P0955
SYM046
SYM014, SYM049A
P0493
P0540
P0767
SYM117A
P0514
P0571, P0692
P0768
P0955
P0353
SYM177
Reference
P0596, P1050
SYM080
P0693
P0439
SYM005, SYM128
P0967
SYM157
SYM062
SYM139
P0125
SYM161
P0198
P0394
P0690
P0369, P0760,
P0761, P0999
SYM067
P0440
P0806
Chiu, Shau-Ting
Chivers, Steve
Cho, I-J
Choat, Brendan
Chobot, V.
Chochois, Vincent
Choi, Byoung-Hee
Choi, Ji-Eun
Choi, Y-S
Chokan, Rajab
Chokchaichamnankit,
Ploenpit
Chong, Caroline
Chookaew, Sirilak
Chotchoungchatchai,
Somtanuek
Chou, C.H.
Choudhary, Ritesh
Christenhusz, Maarten
Christie, Michael
Christman, M.
Christopher, J
Chrtek, Jindrich
Chuan Ho, Boon
Chuang, H.T.
Author
Chen, Yu
Chen, Zhi-Duan
Cheng, Ping-Chin
Chenu, Karine
Chesselet, Pascale
Chesson, Peter
Chester, Michael
Cheung, Alice
Chew, Tania
Chi, Katherine
Chia, Jer-Ming
Chia, Kerryn
Chia, Tet Fatt
Chiang Shiong, Loh
Chiang, Yu-Chung
Chickarmane, Vijay
Chika, Ejima
Chimal-Hernandez,
Aurora
Chimphango, S.
Chiou, Wen-Liang

Author
Chung, Kuo-Fang
Chung, Kyong-Sook
Chye, Sam
Chytrý, Milan
Cibrian, Angelica
Cifuentes-Blanco, J.
Clapham, Alan
Clark, B
Clark, Lynn G.
Clarke, Andrew
Clarke, Laurence
Reference
SYM008, P0769
SYM085
P0470
SYM018
SYM048, SYM119
P0750
SYM013
P1015
P0913
P0254
P0212
Classen-Bockhoff,
Regine
Clayton, Harmony
Clearwater, Michael
Clement, Mathilde
Clements, Mark
SYM076A, P0770,
P0771, P1043, P1044
P0593
P0470
SYM052
SYM057, SYM117,
SYM119
SYM018
P0036, P0090
Clements, Steven
Close, Dugald
Coan, Alessandra
Coates, David
Coates, Fiona
Coelho, G. O.
P0618, P0689
KNS10, SYM009
SYM167
P0191
Coelho, L.F.M.
Coffani-Nunes, Joao
Vicente
Coffin, Douglas W.
Cogan, N O I
Coiffard, Clement
Coissac, E.
Collins, Helen
Collins, Michelle
Colmer, Timothy
Columbus, Travis
Comer, Jason
P0065
P0772
P0660
P0269
SYM006, SYM106B
SYM013
Cole, M. P0389, P0483
Collado-Torres, Ricardo P0057
SYM059, P0259,
P0471
P0125
SYM012, P0569,
P0965
SYM106B, P0976
SYM109
Conant, Gavin
Congdon, Robert
Conn, Barry J.
Conner, Joann
Conran, John G
Consaul, Laurie L.
SYM157
P0035, P0578
SYM099, P0800
SYM039, SYM162
SYM101,
SYM106A,
SYM117, P0729
P0938, P0939
Author
Conti, Elena
Conway, Janine
Cook, Lyn
Cook, Martha
Cooper, Alan
Cooper, Endymion
Cooper, Laurel
Corcuera, Luis
Cordeiro, In¨ºs
Corlett, Richard
Corley, Aaron
Cornelissen, Hans
P0297
SYM018
SYM048
SYM014
Cornwell, William
Correa, Regis
Correa-Gómez, Diego
Felipe
Cortés-Palomec, A.
SYM014, SYM092
SYM060B
SYM164
Cortez, Priscila
Andressa
Coruzzi, Gloria
Corvez, Adèle
Cosse, Mathilde
Costa Penatti, Natasha
Costa, A.C.G
Costa, Daiana
P0886
P0702
SYM048, SYM119
SYM116A
SYM059
P0012
P0106
SYM072
Costa, I.R. SYM131
Costa, Rafael Carvalho P0162
Costion, C.
Costion, Craig
Cottage, Amanda
Couceiro López,
Francisco J
Coudouel, S
Courrech Du Pont,
Sylvain
Courty, Pierre-
Emmanuel
Cousins, Asaph
Reference
SYM072,
SYM102B,
SYM127,
SYM144A, SYM171
P0619
SYM104B
SYM040
SYM013
SYM155
SYM048
P0024
SYM092
SYM088, SYM144B
SYM063
P0408
SYM157
SYM066
SYM124
Coutinho, Italo
Couturier, Etienne
Couvreur, Thomas
Covington, Michael
Covshoff, Sarah
Cowley, Kirsten
Cox, Simon
Cozzolino, Salvatore
Craig, Kimberly
SYM050, P0255
SYM089B, P0117
SYM065, SYM066
SYM109, P1045
SYM045
SYM028
P1046
P0420
SYM002, SYM073,
SYM105, SYM142,
P0926
P1081
693

Author
Crandall-Stotler,
Barbara
Crane, Peter
Craven, Lyn
Crayn, Darren
Creasey, Kate
Creese, Chris
Cremer, Julia
Crespo, Ana
Crespo, E.
Crespo, M.B.
Cress, John
Crisci, Jorge
Crisp, Michael
Crisp, Peter
Cristescu, Simona
Cristofanelli, Paolo
Croft, Jim
Croft, Larry
Crone, Elizabeth E.
Cronn, Richard
Crookshank, Annabel
Cross, Hugh
Cross, R
Crous, Kristine
Cruaud, Corinne
Cruz, Alex
Cruz, Marisol
Cruz, W
Cui, Hong
Cui, Ke-Ming
Cuin, Tracey A.
Cullis, Christopher
Cúneo, Ruben
Cunha, Alexandre
Cunningham, Saul
Curaba, Julien
Curlevski, Nathalie
Cury, Graziela
Cushman, John
Cushman, John C
Cusimano, N
Reference
SYM155
SYM134
SYM060
SYM014
SYM007, SYM038A
SYM088B
SYM013,
SYM088B, P1053
P0164
P0047
P0814
P0114
P0024
SYM011
SYM136, SYM137
P0621
P0231
SYM123
SYM092, SYM101,
SYM121, P0048,
P0820
SYM067
SYM001, SYM078,
SYM126, SYM166
P1101
SYM016
SYM069B
SYM045
SYM045, P0373,
P0374, P0375
SYM106B
KNS10, SYM128
SYM131
SYM004, SYM088,
SYM092, SYM017,
SYM104B,
SYM108, P0026
KNS01
SYM116B
P0592
SYM093
SYM010
P0833, P0834
SYM136
SYM108
PL09, SYM104,
SYM131, P0808
SYM054, SYM063,
P0620
SYM010
P0002
694
Author
Da Cunha, Maura
Da Silva, Clarisse P.
Daehne, Judith
Dafni, Amots
Dai, Xiaofei
Dalton, Heidi
Dalton, PATRICK
Reference
P0613
SYM002
SYM136
SYM142
P0615
SYM053
SYM158, P1000
Dalvi, Valdnéa
Damdinsuren,
Oyunchimeg
Damesin, Claire
Dan Chitwood, Dan
Dane, Feruzan
Danhof, Linda
Darbyshire, Iain
Dart, Sara
P0117
P0597
SYM105A
SYM069B
P0354, P0351, P0359
SYM059
SYM128
SYM102
Daru, Barnabas Haruna P0774
Das, Pijush Kanti P0775
Dassanayak, Maheshi
Daubaras, Remigijus
Dauphinee, Adrian
Davenport, R
SYM041
P0919
P0622
SYM157
Davidson, Amy
Davidson, Neil
Davies, Faith
Davies, William
SYM020
P0036, P0090
SYM048
SYM036
SYM140 D'ávila De Moraes,
Miguel
Davila, Yvonne
Davis, Aaron
Davis, Arthur R.
Davis, Christine
Davis, Jerrold
Davis, Jonathan
Dawson, M.I.
Day, Peter D.
P0233
SYM128
SYM058
SYM155
SYM106A, SYM109
SYM059
SYM108
P0845
Dayod, M SYM049A
De Almeida Vieira,
Fabio
P0037
De Andrade, Guilherme P0656
De Carvalho, Dulcineia P0037, P0038
De Clerck, Olivier
De Coligny, François
De Freitas Mansano,
Vidal
De Groeve, Steven
SYM075
P0612
SYM089B
SYM071
De Jaeger, Geert SYM071
De Jager, Marinus Louis SYM102A

Author
De Kok, Rogier
Reference
SYM166
De La Herrán, Roberto P0407, P0408
De La Paz Perez Olvera,
Carman
De Lange, Peter
SYM001
SYM085, P1071
De Lemos Filho, José
Pires
De Luna, Efrain
P0040
SYM139, P1000
De Melo Bandeira,
Helena
De Menezes, Nanuza
Luiza
De Moor, C.
De Moraes, Miguel
D`Ávila
De Moraes, Pedro Luís
Rodrigues
De Proft, Maurice P
P0012
SYM069B
SYM071
SYM140
P0795
De Quieroz, Luciano
De Reffye, Philippe
De Saint Germain,
Alexandre
De Sousa, F.D.P.S.T.
SYM045
SYM089
SYM066
P0594
SYM074
De Souza, Cacilda
Ad&Eacutelia Sampaio
De Souza, Guilherme
P0142
P0657
De Souza, Pedro Paulo P0232
De Vos, Jurriaan
Deák, Áron József
Dearnaley, John
Debladis, Emilie
SYM102B
P0777
SYM124
SYM162
Deboer, Kathleen
Decombeix, Isabelle
Degtjareva, Galina
Deguchi, Hironori
Dehghani, Gita
Del Castillo, Rafael F.
Delgado Salinas,
Alfonso
Dell, B
Deloger, Marc
SYM053
SYM171
SYM079A, P1032
P0838, P0856,
P0940, P0968, P1000
P0305
P0042, P0197
SYM089A, SYM150
Delpratt, John
Delventhal, Naomi
Delwiche, Charles
Demarco, Diego
Demetras, Nick
Demidenko, Natalia
Dempsey, Raymond
Demyrelma, Hakki
Deng, Zhen-Hai
Denham, Andrew
SYM160
SYM157
P0092
P0783
P0401
P0624
P0798
P1047
P0310
P0785
SYM057
SYM127
Author
Dennetière, B.
Dennis, Elizabeth
Dennis, Liz
Reference
SYM005
SYM123, SYM152
SYM060
Deore, Gopal
Depamphilis, Claude
P0513
SYM038B,
SYM080,
SYM117B,
SYM157, P0377,
P0386
P0732 De-Polli, Bm
Der, Joshua
Derzhavina, Nina
Desalle, Rob
Deshmukh, Yogita
Desnos, Thierry
Dessein, S.
Devall, Margaret
Dever, Louisa V
P0377, P0386
P0626
SYM048, SYM119
P0256
SYM052
SYM137
SYM080
SYM045, P0375,
P0385
SYM115
SYM069B
Devey, Dion
Devisetty, Upendra
Devos, Katrien
Devos, Nicolas
Dewar, Ken
Dexter, K
SYM161
SYM155
SYM096
SYM077B
Dhabe, Arvind
Dhingra, Gulshan
Kumar
Dhugga, Kanwarp
P0627, P0827
SYM163, P0778
P0589
Di Romagnano, Ligia
Ferrari T.
P0292
Dias, Alexandra Soveral SYM003
Dias, Luís Silva
Dias, Mário Sérgio
Carvalho
Diaz, Maria Francisca
P0045
P0366
P0257
Diaz-Castelazo, Cecilia SYM150
Diazgranados, Mauricio SYM168, P0779,
P0822
Diaz-Larrea, Jhoana P0756, P0757
Díaz-Lopez, Hilda
Dibley, K
Dick, Christopher W
Dierschke, Tom
P0057
P0483
SYM114
SYM040,SYM097
Dietrich, Sonia
Dietzel, Lars
Diggle, Pamela K
Dikmen, F.
Dillon, R.
Dillon, S. K.
Dimitroff, George
P0467
SYM063
SYM076A
P0770
SYM009
SYM037
P0475
695

Author
Dinesh-Kumar, S.P
Divakar, Pradeep K
Dixon, Bob
Reference
SYM041
SYM093
SYM167
Dixon, K.W.
Dixon, Kingsley W
Djieto-Lordon,
Champlain
Dkhar, Jeremy
Dludlu, M.
Dlugosch, Katrina
Do Carmo E. Amaral,
Maria
Dobbs, Cynnamon
P0120
SYM153, SYM166,
SYM167, P0154,
P0198
P0208
SYM080
SYM129
SYM025, SYM038
SYM099B
Dobignard, Alain
Doblin, Monika
Docktor, Victoria
Dodd, Ian
Dodds, Peter N.
Dodgson, John
SYM018
SYM118
SYM046, SYM059
SYM117B
SYM036
SYM055
SYM009
Doerge, Rebecca KNS01
Dogan, Hasan Huseyin P0046
Dogan, M.
Dolati Baneh, Hamed
Dolferus, Rudy
Dolja, Valerian V.
P0770
P0237
SYM160
SYM061
Domínguez Lozano, F. SYM167
Domínguez, César A. SYM114
Donaldson, Lara
Dong, Aiwu
SYM153
SYM123
Dong, Shi-Yong
Dong, Yunfa
Dönmez, Ali A.
Donnon, Matthew J.
Donoghue, Michael J
Dornelas, M.C.
Dorr, Laurence J.
Dorsey, Brian
Dötterl, Stefan
Douady, Stéphane
Doust, Andrew
Downie, Stephen R.
Doyle, James A.
Doyle, Jeff J.
Doyle, Vinson
P0760
P0596
P1033, P1084
SYM117
SYM077, SYM086
SYM131
SYM136, P0780
SYM081
SYM105
SYM056, SYM065,
SYM066,SYM067
SYM069, P0378
SYM079A,
SYM079B,
P0412,P0792
SYM099A
SYM074, P0421,
P0809
SYM027
696
Author
Draper Y Diaz De
Atauri, I.
Dreccer, Fernanda
Dreccer, M F
Dresselhaus, Thomas
Dressler, Stefan
Drew, Bryan
Drigo, Barbara
Driscoll, C.
Du, Juan
Du, Yejie
Duan, Qiaohong
Duan, Yizhong
Duangjai, Sutee
Duarte-Silva, Erica
Dubuisson, Jean-Yves
Reference
P0051
Duckett, Jeffrey
Dudash, Michele
Dueck, Lucy
Duffy, Karl
Dun, Elizabeth A
Duncan, M. J.
Duncan, Richard P
Dunn, Christopher
Dunn, Elizabeth
Dunoyer, Patrice
Dutt, Som
Duvall, Melvin R
Dwari, Saurav
Dwivedi, N.K.
E. Valente, Gilmar
SYM127
SYM105B
SYM102B
P0592, P0594
SYM167
SYM018
P1085
SYM153
KNS01
SYM159, P0511
SYM106B
P0784
P0298,P0299
P0232
Eamens, Andrew L
Ebihara, Atsushi
Ebine, Kazuo
Ebner, Martin
Ebrahimzadeh, Hassan
Eckert, Christopher
ECOCHANGE/Barfrost
Teams, A2
Edelin, Claude
SYM060B
SYM095, P0863,
P1029
P0628
SYM151
P0284
SYM102
SYM088
Edelmann, Richard E.
Edens-Meier, Retha
Edger, Patrick P
Edwards, Aaron
P0688
SYM065
SYM142
SYM096, SYM157
SYM053
Edwards, Bort (Robert) SYM131
Edwards, Erika SYM017, P0516
SYM034
SYM160
KNS04
P0781,P0782
P0783
SYM016
P0389
SYM069B
SYM065
SYM062
P0762
SYM168, P0947
P0656
SYM116A,
SYM144B
SYM087

Ellis, Jeffrey G.
Ellis, Miriam
Ellsworth, David
Elser, Justin
Elvira, Hoerandl
Emadzade, Khatere
Emiko, Oguri
Emshwiller, Eve
Endress, Mary
Endress, Peter K.
Engel, John
Enke, Neela
Ennos, R. A.
Ennos, Richard
Epp, LS
Epperson, B.K
Era, Atsuko
Author
Edwards, Everard
Edwards, Gerald E
Reference
P0595
SYM028, SYM068,
SYM127, P0225,
P0696
SYM157 Edwards, Keith
Edwards, Robert
Edwards, Trevor
Edwin-Wosu, Nsirim
Eeckhout, Sharon
Egan, Ashley N.
Egelund, Jack
Egerton, Jack
Ehrendorfer, Friedrich
Ehrlen, Johan
Ehsanpour, Ali Akbar
SYM131
P0930
P0141
SYM069B
SYM074
SYM046
SYM022
SYM094,P0811
SYM014
P0603
Ehteshamnia, Abdollah P0266
Eichhorn, Karl A. O. SYM110
Eini, Omid
Eiserhardt, Wolf L.
Ejima, Chika
El Ottra, Juliana H. L.
P0287,P0380
SYM109
P0440
SYM069A
El-Awadi, Mohamed
El-Bassiony, A.M.
Elbl, Paula Maria
Elfadly, Eslam
Eliby, Serik
Elliott, Carole P
Elliott, Catherine
Ellis, Allan George
SYM031
P0319
SYM069B
SYM023
P0380
SYM018, SYM166
SYM156
SYM102A
SYM055
SYM046
SYM049B, P0047
SYM048
P0074, P0824
P0823
P1065
SYM011
SYM072
SYM099B
SYM118, SYM155
SYM120
SYM077
SYM119
SYM088
SYM011
P0628
Author
Erickson, D L
Eristavi, Marine
Ermawar, Riksfardini
Ernst, Evan
Eronen, Jussi
Ertugrul, Kuddisi
Escapa, Ignacio
Escobar-Restrepo, Juan-
Miguel
Escolastico-Ortíz,
Dennis
Escudero, Marcial
Reference
SYM114
P0121
P0259
KNS01
P0960
P0785
SYM007, P0048
SYM064
P0937
Esfeld, K.
Esmailbegi, Shokouh
Esparza, E.
Esparza, Iris
SYM085, P0786,
P0787
P0928
P0842
P0184
P0796
Espino, Susana
Estavillo, Gonzalo M
SYM049A
SYM054,
SYM063,P0620
P0060 Esteban, Raquel
Estébanez Pérez, Belén P0049, P0050,
P0051, P0788, P1000
Evans, John SYM028, P0263
Everly, Robin
F. Fay, Michael
Faast, Renate
Fabrizio, Jacqueline
Facelli, Jose M.
Faezbakhsh, Alireza
Fagundez, Jaime
Faigl, W.
SYM136
SYM105, P0926
SYM169
SYM059, P0471
SYM001, SYM169
P0539
P0053, P0789
SYM042
Faik, Ahmed
Faithful, Troy
Falat, Igor
Falcinelli, Mario
Falik, Omer
Falkiewicz, Agnieszka
Fan, Jian-Hua
Fang, Wen
Fangmeier, Andreas
Faria Salatino, Maria
Luiza
Faria, Danielle A
Farias-Singer, Rosana
SYM034
P0297
SYM037
P0974
Farinaccio, Maria Ana P0871
Farinha-Marques, Paulo P0790
Farruggia, Frank
Farzamei, Mojgan
P0791
P0567
SYM046
SYM162
P0675
SYM162
SYM020
SYM032
SYM080
SYM008
697

Author
Fawzy, Z.F.
Fay, Michael F.
Fayant, Pierre
Fehrer, Judith
Fei, Xu
Feijo, José
Feild, Taylor
Feist, Mary Ann
Fekete, Megan
Feldberg, Kathrin
Feldman, Aryo
Feliner, Gonzalo N.
Felix, Lucas Macedo
Feng, Chang-Lin
Feng, Dan
Feng, Xu
Feng, Yun
Fenske, Ricarda
Feodorova, Tatiana
Ferjani, Ali
Fernandes, Monique
Carolina Nunes
Fernandez-Marin,
Beatriz
Fernández-Mazuecos,
Mario
Fernández-Palacios,
J.M.
Ferraz, Camila
Ferreira, Juliano Lino
Ferro, Myriam
Fesenko, Alexei
Feuillet, Christian
Fidelis Duarte,
Jaqueline
Fiehn, Oliver
Field, Ashley
Field, David. L.
Field, Katie
Fields, Allison M
Fife, Allan J.
Figueiredo-Ribeiro, Rita
De Cassia
Filado, Fiona
Filer, Denis Leonard
Filgueiras, T.S.
Reference
P0319
SYM002, SYM004,
SYM038B,
SYM105B, P0845,
P1052
SYM065
SYM170, P1049,
P1064
P0350
SYM062
SYM080
P0792
SYM058
SYM155
SYM028
P0910
P0479, P0480
SYM057
P0054, P0764
P0596,P1050
P0068,P0264
P0591
P1051
P0629
P0366
P0592
P0717
P1006
P0060, P0481
SYM167, P0787,
P1052
SYM127
SYM072
P0364, P0365,
P0366, P0367
SYM068
P1047
SYM171
P0037
SYM069B
SYM116B
SYM102B
SYM051
SYM062
SYM158
P0482
698
Author
Fincher, Geoffrey
Finigan, Patrick
Finnegan, Jean
Finnegan, Patrick M.
Fischhoff, David A
Fishbein, Mark
Fisher, Kirsten
Fisher, Nigel
Fitzgerald, Glenn
Fiz-Palacios, Omar
Flann, Christina
Flannery, Maura
Flematti, Gavin R
Fleury, Delphine
Flores-Sandoval,
Eduardo
Flowers, Timothy
Floyd, Sandra
Foden, Wendy
Foley, William
Fontana, S.L.
Fontes-Soares, Bárbara
Dantas
Foo, Eloise
Forczek, Ewa
Ford, Chris
Forest, F
Forest, F.
Forest, Felix
Forni-Martins, E.R.
Fornoni, Juan
Forrest, Cairo N.
Forrest, Laura
Forster, J W
Forster, Paul I.
Fortelius, Mikael
Fortunato, Renée
Fosu-Nyarko, J
Fournel, Jacques
Fournier-Level,
Alexandre
Foyer, Christine
Fozuni, Masoome
Frahm, Jan-Peter
P0484, P0485
P0055
P0448
SYM129
P0816
SYM103, SYM109,
SYM115,
SYM117A,
SYM131, P0737,
P0952
SYM131
SYM114
SYM127
SYM087, SYM155
P0269
SYM072
P0960
P0381
SYM160
SYM057
SYM069A
SYM070
P0237
SYM122
Reference
SYM059, P0259,
P0340, P0425,
P0431, P0471,
P0475, P0515, P0589
KNS01
SYM060,SYM162
SYM041, SYM052
PL02
SYM038A, SYM072
SYM098
P0483
SYM034, P0176,
P0194, P0310
P0787
SYM118,SYM120
SYM005
SYM153, P0066
SYM071
SYM042
SYM041
SYM040
KNS05
SYM037, SYM121
SYM013
P0366

Author
Franc, Alain
Francelino, Márcio
Rocha
Francino, Dayana
Reference
SYM088B
P0153
SYM089B
Francis Roche, Kennedy P0012
Francis, Louise P0952
Francisco, Ana SYM142
Francisco-Ortega, Javier SYM109, P0889
Franco, A. C.
Franklin-Tong, Noni (V.
E.)
Franks, Steven
P0555, P0556
KNS04, SYM070
SYM025, P0211
Franz, Hadacek
Franzaring, Jürgen
Freckleton, Robert
Fredriksson, Gabriella
Freeland, Christopher
Freeman, Jules
Fregonezi, Jeferson
Freihat, Lubna
P0074
SYM034
P0516, P1066
SYM110
SYM134, SYM136
SYM149
P0974
SYM153
Freitas, Loreta
French, Kris
Frérot, Hélène
Freschi, Luciano
Frickey, Tancred
Friedman, William E.
Frigerio, Jean-Marc
Friis, Else Marie
Fritsch, Peter W.
Frodin, David G.
Frole, Kristen
Fromhold, Samantha
Fshar Mohammadian,
Mansoor
Fu, Chengxin
P0974
P0187
SYM171
SYM045, P0601,
P0602
P0592
SYM064, SYM080
SYM088B
PL01
P0147,P0859
P0929
SYM056
P0382
P0462
Fu, Feixue
Fu, Peili
Fu, Pengcheng
Fu, Yu-Lan
P0763, P0793
SYM026
SYM049B
P0762
P0693
Fuentes-Cross, Patricia P1053
Fuglsang, Anja P0231
Fujibayashi, Shigetaka
Fujii, Mutue T.
Fujii, Nobuharu
Fujii, Sota
Fujimoto, Ryo
Fujinami, Rieko
P0643
P0756, P0757
P0522, P0575
SYM055, P0635
SYM123
P0630
Author
Fujita, Asami
Fujita, Minoru
Fukada, Hiroo
Fukuda, Hiroo
Fukuda, Tomoko
Fukusaki, Eiichiro
Fulop, Dan
Funk, Vicki A.
Funnekotter, Bryn
Furbank, Robert T
Furlan, Claudia
Furukawa, Jun
Fusaro, Adriana
Fuse, Shizuka
G.T. Willats, William
Gabedava, Laura
Gaff, Donald
Galán, Pablo
Galla, Giulio
Gallagher, Catherine
Gallagher, Michael
Gallego, Berta
Gallegos, Clemente
Galley, Chloe
Gallwey, Janice
Galtier, J.
Gama, Lilia
Gamage, Harshi
Gamarra, Roberto
Gandhi, Kanchi N
Gandolfo, Maria A.
Gangale, Carmen
Ganhão, E.
Gao, Chien
Gao, Lian-Ming
Gao, Liyan
Gao, Qingbo
Gaquerel, Emmanuel
Garcia Medina, N.
Garcia, Nicolás
García-Castaño, Juan
Luis
Garcia-Plazaola, Jose
Ignacio
Gardiner, Donald M.
Reference
P0397
P0699
P0454
SYM061, P0632
P0794
P0560
SYM069B
SYM120
P0084
SYM028, SYM035,
SYM048, P0540
P0297, P0486
P0499
SYM060B
P1054, P1055, P1072
SYM046
P0122
SYM054, P0465
P0796
SYM162
P0795
SYM128
P0155
P0174
SYM103
SYM076
SYM116A
P0057, P0750
SYM104B
P0796
P1100
SYM048, SYM092,
SYM101, SYM121,
P0820
SYM011, P0753
P0045
P1020, P1021
P0059, P0797
SYM159
P0762
SYM010
P0049, P0050, P0051
P0890
SYM126, P0383,
P0948, P0949, P0993
P0060,P0481
KNS08
699

Gavrilova, Olga
Gaymer, Carlos F.
Ge, Xuejun
Gebauer, Gerhard
Gehring, Chris
Gehrke, Berit Gehrkef
Geiger, Jennifer M. O.
Geisler, Matt
Geitmann, Anja
Geltman, Dmitry
Gemeinholzer, Birgit
Gemmill, Chrissen
Génova Fuster, M.M.
Gentili, Rodolfo
Genung, Mark
Georginah, Molotja
Gerber, Daniel
Gereau, Roy
Gernandt, David
Ghahremaninejad,
Farrokh
Ghalavand, Amir
Ghamkhar, Kioumars
Author
Gardner, Martin
Gardner, Michael
Gardner, Stuart
Garilleti Alvarez, R.
Garnett, J S
Garnica, S.
Gärtner, Georg
Reference
SYM119
P1053
SYM088B
P0051
SYM046
SYM027
SYM018
Garzon, Javier
Gaskett, Anne
Gaskin, J. L.
Gasser, Charles
P0805
SYM057, P0224
P0833
SYM150
Gasson, Peter
Gastal Jr, Cláudio
Viníssius De Senna
Gatica, Alejandro
SYM089B
P0153
P0067
Gautier, Laurent SYM118
Gauvin-Bialecki, Anne SYM057
Ghanati, Faezeh
Ghannoum, Oula
Ghimire, Balkrishna
Ghisalberti, Emilio L
Gianoli, Ernesto
Gibbons, Kerry
P0286
SYM162
P0462, P0487,P0488
SYM016
P0819
SYM153
SYM020
P0800
SYM172
P0188
P0101
SYM124
SYM153
SYM085, SYM091
SYM095
SYM048
SYM065
SYM081
SYM120
P0115, P0798
SYM167
P0002
SYM149
P0061
SYM011
P0062
SYM007, SYM119
SYM130, P0799
700
Gleason, Sean
Glen, Melissa
Glenny, Davic
Glinwood, Robert
Göbbeler, Katrin
Godé, Cécile
Godfray, H C J
Godfree, Robert C
Godts, Christof
Goetghebeur, Paul
Goffinet, Bernard
Goldenberg, Renato
Goldstein, Guillermo
Golz, J.F.
Gómez, David
Gómez, L.
Gonçalves Da Silva,
Cristiane
Goncalves Nazareno,
Alison
Gonsiska, Philip
Author
Gibbs, Adele K.
Gigord, Luc
Gijón López, María C
Gil, André S. B.
Gilbert, Benjamin
Giles, Larry
Gille, Sascha
Gillespie, Kelly M.
Gillespie, Lynn J.
Gilliham, M
Gilpin, Amy-Marie
Gil-Rodríguez, Ma.
Candelária
Giraud, Estelle
Giridhar, P
Girlanda, Orlando
Girón, R.C
Giuliani, Rita
Giulietti Harley, Ana
Maria
Giussani, Liliana M.
Givnish, Thomas
Gladish, Daniel K
Glassop, Donna
Glavas, Katarina
Gleadow, Roslyn
SYM100
SYM004,
SYM116A,SYM117
P0631, P0667
P0312
P0542
SYM016, SYM024,
SYM035, SYM053,
P0263, P0382,P0532
SYM014, P0220
P0803
SYM158
SYM010
P0744
SYM171
P1015
SYM165, P0094
SYM045
SYM091
SYM087, SYM155
P0895
P0588
P0450
P0053
P0610
SYM069B
P0063, P0064, P0065
SYM004
Reference
SYM091
SYM144B
P0407
P0718
P0098
SYM156
SYM046
SYM156
SYM100, P0938,
P0939, P1071
SYM049A
SYM127
P0756, P0757
SYM063, P0620
P0565
SYM065
SYM011
P0255
P0802

Author
Gonzalez, Favio
Gonzalez-Ibarra,
Minerva
Gonzalez-Orozco,
Carlos E
Good, Roger
Goodstein, David
Gorecki, Marta
Gosh, T
Gosney, Benjamin
Gostel, Morgan
Gostin, Irina
Goto, Fumiyuki
Gottschalk, Vanessa
Goubet, P.
Govaerts, R
Govindarajulu, R.
Grace, E.
Grace, Olwen
Gradstein, Robbert
Gradstein, S.R.
Graham, James
Reference
P0804, P0805
P0185, P0806
SYM078, SYM170
Graham, Linda
Graham, Sean W.
Grandbastien, Marie-
Angèle
Grandis, Adriana
Grant, Nicole M.
Grassportal
Development Team,
Grattapaglia, Dario
Gray, John
Gray, Sharon
Green, Peter T.
Green, Roger C.
Gregory, Richard
Greimler, Josef
Grether, Rosaura
Griffith, Patrick
Griffiths, Cara
Griffiths, Erin
Griffiths, Howard
Griffiths, Kathleen
P0009
SYM156, P0230
P1066
PL06, SYM037,
P0423
SYM063
SYM034
P0892
P0254
SYM045, P0374,
P0375, P0385
P0067
SYM089
P0889
SYM054, P0465
P0102
SYM087
SYM073, P0920
Griffon, Sébastien
Grimm, Guido
P0612
SYM072
Grimmelmann, Kerstin SYM097
SYM022
PL06
P0066
P0519
SYM149
SYM115
P0585
P0271
SYM151
P0429
SYM118
SYM038B
P0389
SYM117A
SYM118
SYM172
SYM026, SYM040,
P0029
SYM026, SYM040,
SYM086, P0029
SYM090, SYM119,
SYM122, SYM124,
SYM049B
SYM107, SYM157
Author
Grof, Christopher Pl
Groover, Andrew
Gross, Caroline
Gross-Hardt, Rita
Grossniklaus, Ueli
Reference
SYM028, SYM035,
P0312
SYM069B
P0234
SYM064
SYM039, SYM064,
SYM123
SYM071 Groth, D.
Grudinski, Melanie
Gu, Ying
Guala, Gerald
Guarnieri, Massimo
Gubler, F
Gudrun, Krauss
Gueroult, Michaël
Guerrero Legarreta,
Isabel
Guerrero Villaseñor,
Julián
Guignard, Maïté
SYM072
PL10
SYM134
SYM058, P0665
SYM060A
P0509
SYM151
SYM001
P0408
Guilló, A.
Guimarães, C. D. C
Gujas, Bojan
Gulati, Ashu
Gulati, Ranju
Guler, Necmettin
Gulle, Bahay
Gunale, V.R.
SYM077A
P0834
P0191
SYM153
SYM159
P0492
P0356
SYM045, P0373
P0180
Gunawardena, Arunika SYM070, P0622,
P0653
Gund, Shrikant P0032
Gunn, Bee
Gunner, Simon
Guo, Jianlin
Guo, Qi Rong
P0808
P0809
P0815
P0068, P0264
Guo, Shuiliang
Guo, Weihua
Guo, Yanping
Gurav, Rajaram
P0749, P0810
P0069
P0811, P0825
P0032
Gursanscky, N SYM060B
Gusarova, Galina SYM013, SYM088,
SYM096
Gustafsson, A Lovisa S SYM096
Gustafsson, Mats SYM108
Güsten, Stephanie
Guthrie, Kevin
Guu, Te-Yu
Guy, R.D.
H. Rua, Gabriel
Haas, Jeffrey
P0595
SYM128
P0967
SYM164
P0871
SYM041
701

Hagborg, Anders
Hahn, Karen
Hahn, W.J.
Hahs, Amy
Haig, David
Haigh, A
Haile, J.
Hailey, Luke
Hakata, M.
Halim, Ridzwan
Hall, John
Hall, Michael A.
Hall, Neil
Halling, Roy
Hallsdóttir, Margrét
Halpart, Michal
Halvorsen, R.
Hamachi, Leonardo
Hamant, Olivier
Hamasaki, Ryota
Hameister, S
Hamill, John
Author
Hadacek, F.
Hadacek, Franz
Hadian, Javad
Hadincova, Veroslava
Haerizadeh, Farzad
Haevermans, Agathe
Haevermans, Thomas
Hamilton, Matthew
Hammer, Graeme
Hammer, Ø.
Hamon, Perla
Hamrick, J.L.
Hanada, Atsushi
Hanan, Jim
Hanashiro, Isao
Hand, Melanie
Handa, Shinji
Hang, Yueyu
Hanks, Judith
Hänninen, Heikki
Hansen, Michael
P0179
SYM066
SYM171
P0558, P0572
P0619, P0644
P0277
P0269
P0861
P0815
P0931
SYM022
SYM032
Reference
P0493
P0493
P0266, P0267, P0441
SYM025
SYM060
P0814
SYM081, SYM128,
SYM144B, P0814,
P0877, P1040, P1041
SYM118, SYM158
P1037
P0751
SYM018
SYM064
SYM049B, SYM171
SYM013
SYM089B
P0559
P0571, P0692
P0401, P0751
SYM010
SYM045, P0374,
P0375, P0385
P0995
P0720
P0452
SYM013
SYM045, P0494
SYM065
P0442
SYM082
SYM024, SYM053,
SYM054, P0263,
P0382, P0465
SYM149
P0439
702
Author
Hanson, David
Hao, Guang-You
Hara, Hironori
Reference
SYM040
P0588
P0606
Hara-Nishimura, Ikuko SYM061, SYM070,
P0561
Harder, Lawrence D.
Hardtke, Christian
SYM073, SYM076,
SYM102B
SYM153
Hardy, Giles St. E. J.
Hardy, N.B.
SYM024, P0447,
P0591
SYM104B
Hariadi, Yuda
Haridasan, M.
SYM030
P0555
P0816 Harley, Raymond
Mervyn
Harren, Frans J.M.
Harrington, Mark
SYM010
SYM092,
SYM104B, SYM115
SYM103 Harris, AJ
Harrison, N.
Harrison, Paul
Hartwell, James
SYM077
SYM096
SYM045, P0374,
P0375, P0385
SYM059 Harvey, Andrew
Harvey, Paul P1098
Hasani-Nejad, Maryam SYM009, P1058
Hase, Toshiharu
Hasebe, Mitsuyasu
Hasegawa, Toshihiro
Hashida, Shin-Nosuke
P0566
P0453, P0454
SYM034
P0271
Hashimoto, Kayo
Hashimoto, M.
Hashimoto, Takashi
Hassel, K.
Hatami, Ahmad
Hatsugai, Noriyuki
Hauk, Warren
Haushahn, Tobias
Havran, John
Hawker, N
Hawkes, G
Hawkins, Julie A.
Hayase, Masato
Hayashi, Mai
Hayashi, Yoriko
Hayden, M.
Hayes, Richard
He, Lisi
He, Qiong
P0677
P0524
SYM053
SYM013
SYM009
SYM070
P0817
SYM151
SYM172
SYM060
SYM118
SYM105B,
SYM145, P0737,
P0952
P0523, P0566
P0391, P0392
P0239, P0272,
P0283, P0303
P0393
PL06
P0337
P0818, P1035

Author
He, Zhao-Rong
Headland, Lauren
Hedderson, Terry
Heenan, Peter
Hefer, Charles A.
Hegarty, Matthew
Hegeman, A.D.
Heidari, Reza
Heikinheimo, Hannes
Heinken, Thilo
Heinrichs, Jochen
Hell, Rudiger
Heller, Sascha
Helliwell, Chris
Hellsten, Uffe
Hemingway, Claire
Hemming, Megan N
Henderson, Andrew
Henderson, Marilyn
Henery, Martin Lyle
Hennion, F.
Henry, Robert
Henson, Michael
Henty, Jessica
Henwood, Murray
Heo, Kweon
Hepler, Peter
Herendeen, Patrick S.
Herman, Alexei
Hermant, Marie
Hermsen, Elizabeth J
Hernández Damián, Ana
Lilia
Hernandez, Alvaro
Herrera, Carlos M.
Herrera, Javier
Herron, Margaret
Lucille
Hertweck, Kate
Herzog, Elena
Heslewood, Margaret
Heslop-Harrison, J S
(Pat)
Heuret, Patrick
Hibberd, Julian
Hicks, Leslie
Higashitani, Atsushi
SYM065, P0688
SYM028
SYM069B
P0273
Reference
P0874
SYM069B
SYM098, SYM158
SYM097, P0403
PL06, SYM037
SYM157
SYM035
P0237
P0960
SYM164
SYM155
P0620
SYM077B, P1038
SYM060, P1101
PL06
SYM140
SYM152
SYM077B
SYM059
SYM078
P0072
SYM037,
SYM038A, SYM088
SYM037
SYM061
SYM155, P0800
P0819, P0851
SYM062
SYM089A
SYM006
P0072
SYM101, SYM121,
P0820
P0748
SYM041
SYM015, P0116
P0755
SYM170
SYM117B
P1039
SYM131
SYM038B
Author
Higashiyama, Tetsuya
Higuchi, Kyoko
Higuchi, Masanobu
Higuchi, Niro
Hileman, Lena
Hill, Kathryn Edwina
Hill, Laurence
Hill, R.S.
Hilton, Jason
Hilu, Khidir
Hinden, Hélène
Hinojosa, Luis Felipe
Hipp, Andrew L.
Hiraga, Asahi
Hirai, Regina
Hirakawa, Yuki
Hiramatsu, Michikazu
Hirano, Tomonari
Hirata, R.T.
Hirayama, Yumiko
Hiromi, Tsubota
Hironori, Deguchi
Hisamoto, Yoko
Hiscock, Simon J
Hishiyama, Shojiro
Hislop, M
Ho, Angela
Ho, Simon
Hobbhahn, Nina
Hobern, Donald
Hoch, Peter C.
Hoda, M.H.El-Naggar
Hoebee, Susan E.
Hoegy, Petra
Hoerandl, E.
Hofbauer, Wolfgang
Hofberger, Johannes
Hoffmann, Beate
Hofmann, Rainer
Hojsgaard, D.H.
Holford, P.
Holland, Barbara
Hollingsworth, Peter
Holloway-Phillips,
Meisha-Marika
Reference
PL05
P0535, P0495
P1056
P0142
SYM099A
P0073
P0845
P0073
SYM027
SYM086
SYM005
P0821
SYM085, P0787
P0496
P0932
P0632
P0849
P0239, P0283, P0633
P0140
P1073
P0829, P1065
P1065
P0274
SYM126, SYM157
P0572
SYM104B
SYM052
SYM131
SYM073
PU04
P0822
P0238
SYM015, SYM127,
P0234, P0892
SYM034
SYM039, P0823
SYM018
SYM096
SYM014, SYM153
SYM023, P0244
SYM039
SYM171
P0254, P0420
SYM088A
P0355, P0497
703

Hordijk, C.A.
Horgan-Kobelski, T.
Horiguchi, Gorou
Horisawa, Sakae
Horn, James W.
Horres, Ralf
Hosaka, Kentaro
Hoshino, Takuji
Hoshino, Y.
Hosseini, Horyeh
Hou, Yan
Houben, Andreas
Hovenkamp, Peter H.
Howarth, Dianella
Howell, Katharine
Hoyle, Gemma
Hrmova, Maria
Hsieh, Chang-Fu
Hsu, Chi-Yao
Hsu, Ssu-Wei
Hu, Jinjin
Hu, Shusheng
Hu, Yingkao
Hua, Zhihua
Huang, Bingru
Huang, S.C.
Huang, S.-Q.
Author
Holman,
Holman, Garth
Holst, Bruce
Holst, J
Holtum, Joseph A M
Holzapfel, Sebastian
(Avi)
Honaas, Loren
Honda, Hiroaki
Honda, Kazushige
Hong, Deyuan
Hong, Jeong-Ki
Hong, Joon Ki
Hong-Wa, Cynthia
Hopkins, Della
Hopley, Tara
Hopper, Stephen
Hörandl, Elvira
Reference
SYM119
SYM007
SYM004
SYM031
SYM017,
SYM116B, P0578
SYM009, P0115
SYM108
SYM063
SYM022
P0380
P0034
P0432
P0634
SYM164
SYM004
SYM093
SYM085, P0884
P0633
SYM130
P0825
SYM071
SYM110
P0386
P0443
P0391
SYM172
P0850
P0599, P0703
P0814
SYM088B
SYM165
SYM118, P0154
SYM039, SYM094,
P0074, P0823, P0824
SYM010
SYM020
SYM066, SYM071,
P0629
P0387
SYM081
SYM006
SYM039
SYM062
SYM036
P0999
SYM102A
704
Author
Huang, Sanwen
Huang, Shaobai
Huang, Shing-Fan
Huang, Shuang-Quan
Huang, Tseng-Chieng
Huang, Yongjiang
Huang, Yu-Yuan
Huang, Zhuanhua
Huber, Meret
Huck, Norbert
Hudler, Petra
Hudson, Corey
Huerta, Alfredo J.
Huettel, Bruno
Hufford, Matthew
Hugel, Sylvain
Huggins, Lynne
Hughes, Colin
Hughes, Mark
Huh, Sun Mi
Huisman, John
Hul, Sovanmoly
Humaña, Ana Maria
Humphreys, Aelys M.
Humphreys, Claire
Hunsche, Mauricio
Hunter, Jane
Hunter, Keith
Hunter, Mark D
Hurd, Catriona
Hutchins, David
Huttunen, Sanna
Huygh, W.
Hwang, Eul-Won
Hwang, Hyo Joo
Hwang, Yong Sic
Hwong, Jeen-Lian
Hyodo, Hiromi
Hyvönen, Jaakko
Iakimova, Elena T.
Iba, K.
Ibisch, Pierre
Ibrahim, Mohamed
Ibrahim, Samia
SYM075
P0980
SYM167
SYM084, SYM103
SYM040
SYM049B
SYM048
SYM026
SYM114
SYM026
SYM026
SYM122
SYM091
P0444, P0445
P0456
P0456
Reference
SYM107, P0402
SYM156
P1019
SYM102A
P1019
SYM164
SYM114, P0826
SYM036
P1057
SYM064
P0848
SYM037, SYM157,
P0423
P0498, P0977
P0430
P0961
SYM057
P0103
SYM077B,
SYM089A
P1074
P0456
P0034
P0499
SYM122
SYM010
P0524
SYM004, SYM103
P0500
P0666

Author
Ickert-Bond, Stefanie
M.
Ida, Takashi
Idichi, Kazuhisa
Igarashi, Motoko
Igarashi, Yuta
Iio, A.
Ikeda, Hiroshi
Ikeda, Mayuko
Ikeda, Takefumi
Ilbert, Pascal
Iles, William
Im, Chak Han
Imada, Yusuke
Imaichi, Ryoko
Imam Saadi, Sk. Md
Abu
Imamura, Tomohiro
Inada, Akira
Inatomi, Yuka
Indsto, James O.
Ingle, Priyanka
Ingram, Amanda
Inman-Bamber, G
Inoue, Hiroshi
Inoue, Masahiro
Inukai, Yoshiaki
Ionta, Gretchen
Iqbal, Muhammad
Irving, Helen R.
Isagi, Yuji
Isherwood, Siobhan
Ishii, Hiroshi
Ishizaki, Kimitsune
Ishizaki, Kouyou
Ishizuka, Moriyoshi
Islam, Shahidul
Islam, Sharmin
Ismagul, Ainur
Ismail, Abdelbagi
Isnard, Sandrine
Isobe, Sachiko
Itabashi, Etsuko
Ito, Kanae
Ito, Kikukatsu
Ito, Motomi
P0276, P0329
P0357
P0357
SYM057
P0827
SYM106B
P0290
P0643
P0551, P0606
P0455
P0895
SYM030
SYM153, P0341
P0075, P0076,
P0085, P0089
SYM035
SYM076
SYM040, SYM042,
P0628
P0443
P0142
SYM159
P0465
P0287
SYM012
SYM080, P0804
SYM162
P0635
SYM039
SYM156
P0128, P0135,
P0714, P1008
Reference
SYM119, P0222,
P0893, P0894
SYM076
P0277
P0455
P0438
P0524
P1030
P0391, P0392
P0275
SYM162
SYM090
P0684
P0523
P0630
SYM114
Author
Ito, Nozomi
Ito, Takashi
Ito, Yu
Itoh, Kimiko
Itouga, Misao
Ivanescu, Lacramioara
Ivanova, Alexandra
Reference
P0392
P0535
P1073
P0277
P1000
P0585
SYM170
Ivanova, Aneta
Iwabuchi, Hisakatsu
Iwai, Hiroaki
Iwamoto, Akitoshi
Iwashina, Tsukasa
SYM063
P0357
P0499
P0828
Iwata, Natsuko
Izumi, Shoda
Izumidate, Ryoko
Izuno, Ayako
P0597, P0600,
P0604, P0605, P0607
P0635
P0829
P0828
P0075, P0076
Jabaily, Rachel SYM004, SYM108
Jabthong, Rattanawadee P0664
Jackson, Michael B.
Jackson, P
Jacobsen, Sven-Eric
Jacoby, Richard
SYM012, P1015
P0290
SYM030
SYM156
Jacquemyn, Hans SYM105
Jacques, Frederic M.B. SYM164
Jafari, Farzaneh
Jaffré, Tanguy
P0844
SYM119
Jäger, Anna
Jahufer, Zulfi
Jain, Bhanukumar
Jaiswal, Pankaj
SYM145
P0244
SYM177, P0637
SYM048, P0388
Jaligot, Estelle
James, Elizabeth
James, Shelley
Jamzad, Ziba
Janarthanam, Malapati
Jang, Jun Hyoung
Janka, A
Jankauskaite, Rasa
SYM162
P0077
SYM166, P0078
SYM009, P1058
SYM002
P0214
SYM067
P0226
Jansen, Robert
Jansen, S.
Jansen, Steven
Janssen, Thomas
Janssens, Steven
Jaramillo, Carlos
SYM038B, P0412
SYM049
SYM014, SYM115
SYM137, P0782
SYM069B, SYM115
SYM077B
705

Author
Jardim, Jomar
Jardine, Duncan
Jarošík, Vojtìch
Jasieniuk, Marie
Jean-François, Molino
Jedrusik, Nicole
Jeffares, Lynne
Jeffers, S.S.
Jenjittikul, Thaya
Jenkins, Jerry
Jeon, Youngmun
Jeong, Jihee
Jeong, Mi-Jeong
Jerominek, Markus
Jersáková, Jana
Jet Aw, Sze
Jgenti, Lali
Ji, Wenli
Ji, Xuemei
Jia, Wei
Jiang, Dong-Xun
Jiang, Feng-Qing
Jiang, Yanjuan
Johnson, S.D.
SYM062
P0122
SYM085
SYM160
P0287
P0507
P0207
SYM049B
Jiao, Yuannian
Jigami, Yoshifumi
Jiménez Rodríguez,
Francisco
Jiménez, Iván
SYM157
SYM046
P0889
P0199
Jimenez-Gomez, Jose
Jiménez-Mejías, Pedro
SYM069B
SYM085, P0786,
P0787
P0577, P1017 Jin, Bom Bi
Jin, Xiaohua
Jiping, Xuan
Joaquim, Emanuela O.
Jobling, Steve
Jobson, Richard
Johanna, Schmitt
P1078
P0350
P0482
SYM059
P0831
SYM069A
Johanna, Wagner P0824
John De Britto, Alexsis SYM163, P0080
Johnson, Alexander
Johnson, Cory
Johnson, Eric
Johnson, Gabriel
Johnson, K.
SYM035, SYM159
SYM123
SYM062
SYM100
SYM092, SYM101,
SYM121, P0820
P0832
Reference
SYM076B
SYM088B
SYM018
SYM089B
P0740
P0443
P0921
P0894
SYM177
PL06
P0851
P0069
P0444, P0445, P0514
P1044
SYM105A, P0832
706
Author
Johnson, Steven
Johnson, Susan D.
Johnston, Peter
Johson, Alexander
Joly, Alexis
Joly, C.A.
Joly, Carlos
Joly, Simon
Joly-Lopez, Zoe
Jonas, Rayko
Jones, Anne
Jones, Cynthia S.
Jones, Michael
Jones, R.H.
Jones, Rebecca C.
Jones, Samuel S
Jönsson, Henrik
Jordan, Greg
Jordon-Thaden, Ingrid
Jørgensen, Kirsten
Joshi, Bhushan
Joshi, Jasmin
Jost, Ricarda
SYM067
SYM014,
SYM049B,
SYM056, SYM092,
SYM104B
SYM157
SYM053
P0157
SYM164
SYM041, SYM052,
P0591
SYM037 Joubert, Fourie
Joubert, Lize
Jouhet, Juliette
Jourdan, Christophe
Joyeux, M
Juan, Ana
Judd, Walter
SYM168
SYM063
SYM067
SYM151
P0833, P0834, P1052
P0895
Judice, Diogo Marcilio P0081, P0082, P0083
Judzentiene, Asta P0449
Juhász, Ana Cristina
Jullien, Pauline E.
Jun Bgi, Wang
Juranic, Martina
P0366
KNS01
SYM161
KNS04
Juretic, Nikoleta
Jürgens, A.
Justin, Preece
K. Bull-Herenu,
Kabouw, Patrick
Kabshawi, Maher
Kaczmarczyk, Anja
SYM069, SYM069A
P0832
SYM048
SYM076
SYM036
P0169
P0084
Reference
SYM057, SYM058,
SYM073,
SYM102B, SYM139
SYM039, P1064
SYM093
SYM159
P0740
P0150
P0167
P0403
SYM069, SYM069A
SYM027
P0389
SYM049, SYM049A
P0282
P0982
SYM121
SYM106B

Author
Kagami, Hiroyo
Kägi, Christina
Kaiser, B N
Kaiser, Pascal
Kajala, Kaisa
Kajikawa, Masataka
Kajimoto, Takuya
Kakishima, S.
Kakishima, Satoshi
Kakizaki, Yusuke
Kakugawa, Yoko
Kakutani, Tetsuji
Kalamei, Zeinab
Kalllunki, Jacquelyn
Kam, J
Kamatsuki, Kaori
Kamisaka, Seiichiro
Kanda, Yui
Kandasamy,
Muthugapatti
Kane, Nolan
Kanehara, Rie
P0361
P0642
P0560
SYM061
Kasperek, Gerwin
Kassim, Abd. Rahman
Bin
KNS02, SYM120
P0391
Kaneko, Shingo
Kaneko, Yukio
Kaneta, Tsuyoshi
Kannangara, Rubini
Kanno, Satomi
Kao, Teh-Hui
Kaplan, Zdenek
Karahara, Ichirou
Karamian, Roya
Karen Ruiz
Bohórquez3, Liz
Karimaei, Sadegh
P0075, P0076,
P0085, P0089
P0605
P0496
SYM053
SYM052
SYM062
P1049
P0642, P0643
P0536
SYM089B
Karki, Jhamak B.
Karki, Shanta
Karlsdóttir, Lilja
Karol, Kenneth
Karron, Jeffrey
Kartawinata, Kuswata
Kasahara, Haruo
Kasahara, Hirokazu
P0644
P0097
SYM028
P0720
P0401
SYM015
SYM110
P0642
P0642
Kasahara, Hiroyuki P0572
Kashiwabara, Kousuke P0885
SYM136
P0172
P0835
P0835
SYM156
P1026
P0428
P0567
P0997
SYM160
Reference
P0397
SYM064
SYM049, SYM049A
SYM126
P0503
SYM053
P0142
Author
Kasuya, Maria
Kasuya, Nobuhiko
Katagiri, Tomoyuki
Katase, Chikara
Katayama, Natsu
Katinas, Liliana
Kato, Hidetoshi
Reference
P0020, P0148
P0275
P0838
P0828
P0839
SYM108
P0983, P1001, P1026
Kato, Hirotaka
Kato, Juntaro
Kato, Keiko
Kato, Masahiro
SYM042
P0391, P0392
P0075, P0089
P0093, P1078,
P0645, P0839, P0863
P0554 Katsuno, Fumiko
Kaulfuss, Uwe
Kautz, S
Kavanagh, Tony
Kaveh, Akram
Kawabe, Akira
Kawade, Kensuke
Kawaguchi, Yu
Kawahara, Takayuki
P0729
SYM035
SYM063
SYM130
P0428
SYM066
P0535
P0840
Kawai-Yamada, Maki P0271
Kawashima, Tomokazu SYM062
Kawauchi, Takayuki
Kawchuk, Lawrence
Kawoosa, Tabasum
Kay, Pippa
P0443
SYM060B
SYM159
P0393
Kazama, Tomohiko
Kazama, Y.
Kazama, Yusuke
Kazan, Kemal
Kazandjian, Aniuska
Kazemi, B
Kazempour Osaloo,
Shahrokh
P0635
P0239
P0283
KNS08
P0841
P0529,P0530
SYM130, P0284,
P0842, P0843 P0844,
P1070
P0032
SYM160, SYM161
Kedage, Vinayak
Keeble-Gagnere,
Gabriel
Keegstra, Kenneth
Keeler, Kathleen H.
Keeley, Sterling
Keeton, Karina
Kelch, D
Kelch, Dean
Keller, Barbara
Keller, P. A.
SYM059
SYM150
SYM108
P0090
SYM119
SYM007
SYM102B
SYM023
707

Author
Kellog, Elizabeth
Kelly, Dave
Kelly, Laura
Kelso, Sylvia
Kenicer, Gregory
Keniry, Kerry
Kennedy, Gavin
Kerbauy, Gilberto
Kessler, Michael
Keßler, Paul J.A.
Kessler, Sharon
Kessler, Sharon
Keszei, Andras
Khalesro, Shiva
Khalighi-Sigaroodi,
Farahnaz
Khamvarn, Vararas
Khan, Faheema
Khanduri, Priyanka
Khavari-Nejad, R.A
Kheffache, Rafik
Khew, Gillian
Kholdebarin, Bahman
Khurana, Parul
Khutsishvili, Manana
Kibble, Natalie
Kidner, C. A.
Kiehn, Michael
Kieliszewski, Marcia
Kienath, Nana
Kikodze, Davit
Kilian, Andrzej
Kim, B-G
Kim, Dong-Kap
Kim, Dool Yi
Kim, Enhye
Kim, Jin A
Kim, Jong-Hwa
Kim, Jung Sun
Kim, Jung Sung
Kim, Nam-Soo
Kim, Sangtae
Kim, Sunghwan
Kim, Sun-Hyung
Kim, S-Y
P0353
P0563
SYM064
P0529, P0530
SYM065
P0394
P0584
SYM061
P0121
SYM059
SYM077
SYM082, P0848
SYM046
SYM064
P0121
SYM037, P0423
P0514
SYM117, SYM117A
P0456
P0069
P0599, P0703
SYM117A, P0214,
P0395, P0849, P0850
P0599, P0703
P0850
P0395, P0505, P0849
P0851
P0069
P0505
P0514
Reference
SYM076B
SYM001
SYM038B, P0845
SYM102B
SYM089
SYM016
SYM048
P0479, P0480
SYM027
SYM104, SYM110,
SYM115, P1022
SYM039
SYM064
SYM121
P0286, P0517
SYM177
708
SYM068
SYM109
SYM062
P0438
P0600, P0604
SYM144B
P0271, P0397
P1015
P0446
SYM168
P0353
SYM093, P0853
SYM089B
P0952
P0990, P0991
P1032
Reference
P0599
P0069
P0438
SYM069B
P0062
SYM022, P0091
P0676
SYM050
SYM120
P0669
SYM072, P0852
P0507
SYM076B
SYM036
SYM065
SYM117, SYM117A
KNS09, SYM136
P0385
SYM045, P0375
SYM078
P0854
SYM084
P1071
P0121
P0428, P0522
P0855
P0274
SYM055, SYM060
P0852
Author
Kim, Woe Yeon
Kim, Zinsuh
Kimiko, Itoh
Kimura, Seisuke
Kindeketa, W.J.
King, Diana
King, Emma
King, Jenny
King, Matt
Kinoshita, Atsuko
Kinoshita, Luiza
Kinoshita, Takeshi
Kirchoff, Bruce
Kirkegaard, J A
Kiss, John Z.
Kiss, Vladimir
Kissling, W.D.
Kita, Daniel
Kitajima, Aya
Kitajima, Junichi
Kitalong, Ann
Kitazaki, Kazuyoshi
Kitching, I J
Kiyohara, Shunsuke
Klak, Cornelia
Klakhaeng, Kanchana
Klazenga, Niels
Kleist, Annabelle
Klitgaard, Bente B.
Kljuykov, Eugene
Kljuykov, Eugeny
Klopper, Ronell
Knapp, Sandra
Knerova, J
Knerova, Jana
Knerr, Nunzio
Knopf, Patrick
Knox, E.B.
Koba, Hidehisa
Kobakhidze, Lia
Kobayashi, Akie
Kobayashi, Makiko
Kobayashi, Mikio
Kobe, Bostjan
Koch, I.

Konarzewski, Tara
Kondo, Katsuhiko
Kondo, Maki
Kondo, Y,
Konishi, Mineko
Konrad, Wilfried
Konrat, Matt Von
Konstantinova,
Alexandra I.
Kooperberg, Rick
Kooyman, Robert
Michael
Koptur, Suzanne
Korall, Petra
Korbecka, Grayna
Korte, Nikola
Kos, Martine
Koschella, Amy
Koshiba, Tomokazu
Köster, Nils
Kosugi, Issei
Koteyeva, Nouria
Koteyeva, Nuria K
Kotina, Ekaterina
Koumoto, Yasuko
Koumoutsakos, Petros
Kovalchuk, Nataliya
Kovarik, Ales
Author
Koch, John
Koch, Kerstin
Koch, Marcus
Kochova, P
Kocyan, Alexander
Kodym, Andrea
Koehler, Angela
Koehler, P.
Koenig, Dan
Koh, Seok Chan
Kohchi, Takayuki
Koi, S
Koi, Satoshi
Kojima, M.
Kokubugata, Goro
Kollehn, Daniel
Koltunow, Anna M.G.
SYM018
SYM092
SYM150
SYM116
P0383
SYM002
SYM036
P0448
P0575
P0095
P0856
SYM068, SYM170,
P0255, P0646
SYM127, P0696
SYM079, SYM079A
P0561
SYM067
P0287
SYM107, SYM157
Reference
P0198
SYM151
SYM097
SYM067
SYM117B
P0092
SYM058
SYM034
SYM069B
P0508
SYM040, SYM042,
P0628
P0839
P0645
P0524
P0093
P0447
SYM039, SYM060,
SYM060A, P1064
P0094
P0597
P0561
P0632
P0669
SYM151
SYM158
SYM079
Author
Kozovits, Alessandra
Krajníková, Eva
Krak, Karol
Reference
P0196
SYM105, SYM105A
SYM170, P0880
Kramer, Andrea SYM140
Krapp, Florian SYM004, P0430,
P1010
Krasovskaya, Liudmila P0650, P0651
Krause, C. SYM027
Krauss, Gerd-Joachim
Krauss, S
Krauss, Siegy
Kravkina, Irina
P0509
P0743
SYM015
P0646
Kreft, Holger
Kreier, Hans-Peter
SYM144,
SYM144A,
SYM144B
SYM155
Kress, W. John KNS09, SYM102,
SYM114
Kretzschmar, Fernanda P0467
Krishnan S., Gopala P0298
Krishnasamy, Gopinath SYM177
Kroeger, Jens
Krohn, Nadia
SYM065
KNS04
Kron, Kathleen A.
Kroon, Johan
Kropf, M
Krosnick, Shawn
SYM168, P0859
SYM032
SYM082
SYM150, P1060
Kruger, Nicholas J.
Ku, Shin-Ming
Kubátová, Barbora
Kubo, Harumori
Kubo, Tomohiko
Kudo, Gaku
Kuhlman, Alyse
Kulheim, Carsten
SYM156
SYM008
SYM105, SYM105A
P0861, P1000
P0397
SYM076
P0722
SYM037
Kumar, Anand Raj
Kumar, Arun
Kumar, Ashish
Kumar, P.
Kumar, Ravi
Kumar, Sanjay
Kumaran, K.P.N.
Kumaria, Suman
Kunkel, Joseph
Kunwar, Ripu M.
Kunzmann, Lutz
Kuo, Li-Yaung
SYM037
P0511
P0157
P0686
SYM069B
SYM159, SYM163,
P0511, P0778
P0180
SYM080
SYM062
P0097
SYM007
SYM095, P0863
709

Author Reference
Kupcinskiene, Eugenija P0226, P0449
Kurokawa, K
Kurosaki, Nobuhira
P0679
P1055
Kusaba, Makoto
Kusano, H.
Kusumi, K.
Kutsuna, Natsumaro
P0451
P0559
P0524
SYM061
Kvacek, Jiri
Kwezi, Lusisizwe
Kwiatkowska, Dorota
Kwon, T-R
Kyozuka, Junko
Kyriacou, B.
Labandeira, Conrad C
Labat, Jean-Noël
P0398, P0864
SYM153
SYM065
P0514
SYM152
SYM035
SYM027
SYM005, SYM128
Labiak, Paulo
Lacroix, Christian
Ladiges, Pauline
P0931, P0932
P0701
SYM089B,
SYM104B,
SYM121, P0077
SYM001 Ladley, Jenny
Lahnstein, Jelle
Lakmini, Robert
Lakshmanan, Prakash
SYM059, P0471
SYM034
SYM030, SYM031,
P0290
P0211 Lalchan, R.
Lam, Hon-Ming
Lam, Vivienne Ka Yee SYM049B,
SYM119, SYM124
Lamb, Jenny P0803
Lambers, Hans
Lameiras, José Miguel
Lanan, Michele
SYM038B
SYM041, SYM052,
P0130
P0790
P0882
Lander, Nicholas
Landherr, Lena
Landrein, Benoit
Lane, Brendan
P1069
P0386
SYM065
SYM067
Lang, Annick
Lang, Daniel
SYM098
SYM048
Langkilde-Lauesen, Ane SYM032
Langridge, Peter P0287, P0326, P0380
La-Ongsri, Woranuch
Lara Garcia, F.
P0352
P0051
Lara-Cabrera, Sabina I. P0733
Larridon, I. SYM091
Larridon, Isabel
Larsen, Stuart
Larsson, Sonny
SYM091
SYM026
SYM145, P0512
710
Author
Laska, A.
Laufs, Patrick
Laumonier, Yves
Lavin, M
Lavin, Matt
Lavin, Matt
Law, Simon
Reference
SYM010
SYM066
SYM110
SYM077B, SYM129
SYM089
SYM089A
SYM063
Laware, Shankar
Lawless, K.
Lawrie, A
Le Brocque, A.F.
Le Comber, Steven C
Le Péchon, Timothée
Le Roux, Marianne
Leake, Jonathan
Leakey, Andrew
P0513
P0393
P0164, P0935
SYM124
SYM157
SYM144B
SYM129
SYM040, SYM051
SYM034, SYM050,
SYM156
P0038 Leao Bandeira De Melo,
Daniel
Lebeda, Ales
Lebel, Teresa
Lechowicz, Martin
P1088
SYM093
P0098
Lee, C.
Lee, Chang Shook
Lee, Chunghee
Lee, Daphne E.
SYM090
P1078
P0819
SYM101, SYM106,
SYM106A, P0729
P0767
P0450
Lee, J. H.
Lee, Joanne
Lee, Joongku
Lee, Jungho
Lee, Min-Joon
Lee, Nam Sook
Lee, Seong-Kon
Lee, Soo In
Lee, Sung-Il
Lee, Y.I.
Lee, Yeon-Hee
Lee, Yi-Chen
SYM094B, P0763
P0215
P0421
P1054, P1078
P0444, P0445, P0514
P0703
P0395, P0505
P0999
P0599, P0703
SYM063
Leebens-Mack, James H SYM106, SYM109,
SYM117, SYM117B
Leegood, Richard SYM028
Legalley, Erin P0498
Léger, Patrick
Lehmann, C
Lehnebach, Carlos
Lehnert, Marcus
SYM088B
SYM014
SYM083
SYM027

Lemos-Filho, J.P.
Lemson, K.L.
Lemson, Kristina
Lenhard, M.
Lenhard, Michael
Lenne, Thomas
Lens, Frederic
Leong, Wai-Chao
Lepedus, Hrvoje
Lepis, K.B.
Lepschi, Brendan
Letelier, Luis E.
Letort, Véronique
Leung, Hei
Leventer, Sinem
Levichev, Igor
Levin, G.A.
Levin, Geoffrey
Levin, Rachel
Lewis, Gwilym
Author
Lehnert, Marcus
Lehtonen, Samuli
Lehvävirta, Susanna
Lei, Mingguang
Lei, Thomas
Leitão, Carlos André
Leitch, A. R.
Leitch, Andrew R
Leitch, Ilia J.
Leite, Daniel Carlos
Leite, Mariangela
Leliaert, Frederik
Leljac-Levanic, Dunja
Lembicz, Marlena
Leme, Elton M C
Lewis, J.D.
Lewis, Louise Ann
Lewis, Paul
Lexer, Christian
Li, De-Zhu
Li, Fay-Wei
Li, Fuzhong
Li, J
Li, Libo
Reference
P0400
SYM116B
SYM082
SYM052
P0840
P0648
SYM038B
SYM107, SYM157
SYM038B, P0845
P0292
P0196
SYM075
KNS04
P0021
SYM004, P1038
P0040
SYM108
P1061
SYM071
SYM071
SYM022
SYM049, SYM069B
SYM008
P0542
P0746
P1046
P0188
SYM066
SYM028
P0351
P0649, P0650, P0651
P0868
P0868
SYM102B, SYM107
SYM102,
SYM102A,
SYM089, SYM089A
SYM049B
SYM086, P0401
P0401
SYM002, SYM004,
P0926
SYM136, P0059,
P0797
SYM088B
P0205
SYM060,
SYM060A, SYM119
SYM155
Lim, Chung-Lu
Lim, T.M.
Lima, Aline Lopes E
Lima, Letícia
Lima, O
Limaye, Abhijit
Lin, Chia-Yu
Lin, Ching-Ping
Lin, Deshu
Lin, Kui
Lin, L
Lin, Q.
Lin, Y
Lin, Yi-Huei
Linde, Celeste
Lindenmayer, David
Author
Li, Ling
Li, Pan
Li, Pan
Li, Qing-Jun
Li, Rong
Li, Rui
Li, Ruiqing
Li, Shu
Li, Wan-Ching
Li, Wei
Li, Xia
Li, Yanhua
Li, Yinhu
Li, Yuan-Fang
Li, Z. H.
Li, Zhen
Li, Zhimin
Li, Zhongyi
Li, Zuhuang
Liang, Dacheng
Liang, Yan
Liang-Sheng, Wang
Liao, Wanjin
P0402
P0984
P1101
P0869
SYM060B
SYM046
P0607
P0402
Liddell, Andrew
Liebel, Heiko
P0717
SYM124
Liede-Schumann, Sigrid SYM072, P0871
Ligavha-Mbelengwa,
Maanda
Ligrone, Roberto
P0061, P0099
SYM087
Lim, Chang Kun P0577, P1017
SYM062
P0766
P1021
P0870
SYM061
P0762
SYM048
P0869
Reference
P0291
P0763
P0793
SYM102, P1043
P1012
SYM059
SYM107
P0967
P0686
P0100, P0292
P0872
SYM157
P0513
SYM008
P0432
SYM061
P0402
SYM060, SYM060A
P0686
SYM060, SYM060A
P1019
P0920
SYM166
711

Author
Linder, Peter
Reference
SYM014,SYM084,S
YM103, SYM139,
P1057
SYM058 Lindgren, Khrystyne
Liparini, Olinto
Liston, A.
Liston, Aaron
Litt, Amy
Little, Alan
P0020
SYM038,SYM038B,
SYM119
SYM007,
SYM038A,SYM038
B
SYM099B,SYM107,
P0211,P0475,P0515
SYM059, P0340,
P0515
SYM007, SYM119
P0103
Little, Damon P
Little, Lorna
Liu, Daping
Liu, Dong
Liu, G.D.
Liu, Hong
Liu, Ho-Yih
Liu, Hui
Liu, Huiliang
Liu, Jie
Liu, Lifeng
Liu, Mei
Liu, Ming-Che
Liu, Qing
Liu, S
Liu, Xiaofeng
Liu, Xin
Liu, Yan
Liu, Yang
Liu, Yea-Chen
Liu, Yidan
Livshultz, Tatyana
Lledó, M.D.
Llugany, Mercè
Lockhart, Peter
P1009
SYM052
P0764
SYM057
P0760
P0516
P0236
P0059, P0797
SYM059
P0792
P0652
P0101
SYM063
P1050
SYM038B
SYM008
SYM155
SYM095,P0874
SYM052
SYM072
P0834
SYM024
Loew, Markus
Logacheva, Maria
Logan, B.A.
Loh, C.S.
Lohmann, L.G.
Löhne, Cornelia
SYM094B,SYM097,
P0420
P0310
SYM090,P1047
SYM049B
P0686
P0747
SYM082
Loizeau, Pierre-André SYM118
Lombardi, Julio Antonio P0875
Long, David
Long, Rowena
SYM155
P0066,P0102
712
Lowry, Iii, Porter P.
Lowry, Porter
Lu, Pei-Luen
Lucas, Eve
Lucas, Michael
Luceño, Modesto
Lucieer, Arko
Luckow, Melissa A.
Ludwig, Martha
Luginina, Ekaterina
Lu-Irving, Patricia
Luk, Michelle
Lumbsch, H. Thorsten
Lundell, Robin
Lundqvist, Lars
Luo, Ming
Luo, Yan-Jiang
Luo, Yi-Bo
Luo, Zhiwei
Luro, Scott
Luthardt, Vera
Luther, Harry
Author
Longhi-Wagner, Hilda
M.
Lonhienne, T.G. A.
Lopato, Sergiy
Lopes, A.V.
Lopez Villalobos,
Adriana
López, A.M.
Lopez, Patricio
Lopez-Bautista, Juan
Lord, Christina
Lord, Janice
Lorence, David
Lou, Yunxia
Loughlin, Patrick
Louis, Cindrella
Lousada, R. B.
Loutfy, Naglaa
Louzada, Rafael
Lovato, Maria
Bernadete
Lowe, Andrew
Lowry Ii, Porter P
Reference
P0913
SYM031,SYM060
P0287,P0380
P0133
SYM102
SYM045
SYM094B,P0067
P0401
SYM070, P0653
P0103
SYM144B
P0810
SYM052
P0547
SYM004
P0551
SYM002, SYM004
P0875,P0104
SYM007,SYM088,S
YM088B,SYM119,P
1053
SYM079,SYM115,P
0722,P0814,P0877,P
0929
P0953
SYM115,P0798,P09
61
SYM009
SYM131, P0737
SYM172
SYM085,P0786,
P0787
P0091
SYM089,SYM150
SYM041,SYM050,P
0593
P0105
SYM077A,SYM077
P0156
SYM093,P0915
SYM022
P0987
SYM060A,SYM060
P1043
SYM057,SYM142
P0470
P0452
SYM164
SYM004

Author
Ly, Ngoc Sam
Lyon, Aidan
Lyon, Stephanie
Lysak, Martin
Reference
P0877
KNS07
SYM105B
SYM096,SYM097,P
0403
P0232 M. Moura, Vitor
M. Schneeweiss, Gerald SYM038B
M.C. Leme, Elton SYM077B
M.G.Pimenov,
Ma, Hong
Ma, Jinshuang
Ma, Shisong
P0706
P0377
SYM165
SYM041
Ma, Wujun SYM159
Maassoumi, Ali Asghar P1070
Mabala, Grace
Mabberley, D.J.
P1080
SYM171
Mable, Barbara
Macas, J.
SYM126
SYM038B,SYM107,
SYM157
SYM171 Macdougal, John.M
Macedo, C.C.L.
Macfarlane, T.D.
Machado, I.C.
P0140
SYM090,SYM117,P
1069
P0106,P0107, P0131,
P0133,P0132
P0106,P0107 Machado, Isabel
Mack, Richard N
Mackay, Alex
Mackay, Duncan
Mackill, David
SYM025
SYM030
SYM150,P0212
SYM012
Mackinder, Barbara A. SYM089A,SYM089
Macklin, J SYM134
Macklin, James
Macphail, Mike
Madadi, L.M.
Madero Pérez, Jesús
Madritch, Michael
Madsen, Leonardo
Maeda, Masato
Maeshima, Masayoshi
SYM137
SYM131
P0062
P0408
SYM149
P0407
P0387
P0629
Magallon, Susana
Magee, Anthony
Magill, Robert
Mahajan, Chetan
SYM086
SYM079A,SYM079,
P0792
SYM118, SYM134,
P0722
P0157
Mahajan, Dnyaneshwar SYM018,P0204
Mahajan, Monika
Mahdavi, Batool
SYM159
P0517,P0286
Author
Mahelka, Vaclav
Mahendran, Vikrant
Mahmooduzzafar,
Mahmoudi Otaghvari -
Kelij, Arman - Sedigheh
Mai, Dieter Hans
Maity, Tapan Kumar
Reference
SYM025, P0113
P0157
P0563
P0109,P0878
Majd, Ahmad
Maki, Jennifer
Malahy, Michael
Malamy, Jocelyn
Malécot, Valéry
Maloof, Julin
Malysheva, Natalia V.
Mamesaya, Ryota
SYM007
P0518,P0519
P0568
SYM058
P0378
SYM046
SYM129,P0879
SYM069B
Manafzadeh, Sara
Mancera, Ricardo L.
Mandak, Bohumil
Mandakova, Terezie
Mandal, Asim
Mandal, Sudhendu
Manley, J.L.
Manners, John M
Mannila, Heikki
Manning, J.C.
P0111
P0606
SYM171
P0084
SYM025,
P0113,P0880
P0403
P0881
P0654
SYM071
KNS08,P0312
Mansor, Asyraf
Mapes, G
Maps, Gene
Marazzi, Brigitte
P0960
SYM117
SYM110
SYM119
SYM007
SYM150,P0882,P08
83
P0655
SYM162
Marc, Jan
Marconi, Gianpiero
Mardonovich, Sandra
Marhold, Karol
Mariath, Jorge Ernesto
Marinho, Cristina
Ribeiro
Marino, Paul
Marjokorpi, Antti
P0421
SYM097
P0656,P0657,P0658
P0704
Markelz, R.J. Cody
Marmottant, P
Maroofi, Hosein
Marques, Isabel
P0224
SYM110
SYM050
SYM151
SYM009
P0721
Marshall, David
Marshall, Mhairi
SYM059
KNS08
Martienssen, Robert A. KNS01,SYM048,SY
M119
Martin, Antony SYM028,SYM035
713

Author
Martin, D
Martin, Noland
Martín, Sandra
Reference
SYM104B
P0986
SYM024
Martin, Tara
Martin, Tarryn
KNS08
SYM050
Martín-Bravo, Santiago SYM085,P0786,P07
87
Martine, Chrstopher T P0951
Martinelli, Gustavo
Martínez, J.
P0081,P0082,P0083
P0184
Martinez, Karina P0188
Martínez-Cabrera, Hugo SYM049A,SYM049
Martínez-Flores, F.
Martínez-Millán,
Marcela
Martinez-Ramirez, J.
P0833
P0758
Martins, Eline
Martins, Euder
Martins, Fernando
Roberto
Marton, Mihaela L
P1068
P0081,P0082,P0083
P0114
P0901
Martos, Florent
Martos, Soledad
KNS04
SYM124
P0155
Masagod, Gayatri P0296
Masakapalli, Shyam K. SYM156
Masaki, Derek
Masaki, Tomomi
Masgod
Chennaveeraiah, Gayatri
Masgod, Gayatri
P0078
SYM085,P0884
P0313
Mason, Michael
Masselon, Christophe
P0296
SYM153, P0592
SYM068
Masselter, Tom SYM151
Mastroberti, Alexandra P0658
Masuda, Akihide
Masumoto, I.
Masuzaki, Hiroaki
Mateo, Patricia
SYM040
P0075
P1000
P0053
Matetovici, I.
Mather, Diane
Mathesius, Ulrike
Mathews, Sarah
Mathieu, Daniel
Mathur, B.K.
Matos, Liss
Matsubayashi,
Yoshikatsu
Matsuda, Ayano
Matsuda, Shinya
SYM013
P0431
P1098
SYM007, SYM119
P0740
P0299
P0297
KNS03
P0392
P0606
714
Author
Matsumoto, K.
Reference
P0552
Matsumoto, Midori P0885, P1028
Matsumura, Aline Tiemi P0520,P0521
Matsuoka, Ken
Matthew, L
Matthews, Merran L.
Matthews, Peter J.
SYM046
SYM060
SYM099B
P0254
Matthews, Sarah
Mattick, John S
Matusick, George
Maurin, Olivier
Mauro-Herrera,
Margarita
Mavrodiev, Evgeny
SYM119
SYM060
SYM024
P0774
P0378
May, T.W.
May, Tom
Mayo, G
Mayo, S J
Mayrose, Itay
Mazhar, Flor
P0979
SYM093, P0982
SYM093
SYM049A,SYM049
P1015
P0787
SYM031
Mazimpaka Nibarere, V. P0051
Mazine Capelo, Fiorella SYM131
Mbayngone, Elisée
Mcadam, Scott Alistair
Mildon
Mccarthy, Michael
P1037
SYM116B
SYM018
Mccauley, Ross P0886
Mccombie, W. Richard SYM119, P0386
Mccomish, Bennet
Mccorkle, Erin
Mccourt, Richard
Mccurdy, David
Mcdaniel, Stuart
Mcdonald, Allison
Mcdonald, Bruce
Mcdonald, Megan
P0420
SYM006
P0401
SYM061
SYM098
SYM156
KNS08
KNS08
Mcdonald-Madden, Eve KNS08
Mcdonnell, Mark SYM018
Mcdougall, Keith
Mcelrone, Andrew
P0077
SYM049A,SYM049
Mcevoy, R.
Mcgee, Peter
Mcintosh, Emma
SYM037
P0129
SYM014,SYM022,
P1062
SYM160 Mcintyre, C L
Mcintyre, Lynne
Mckain, Michael R
SYM160
SYM117B

Author
Mckey, Doyle
Mckinnon, Gay E.
Mckown, Athena
Mclay, T.B.
Reference
SYM035,P0208
SYM121
SYM056,SYM164
SYM104A,
SYM104,P0115
P0439 Mclean, Greg
Mcleish, Michael SYM078
Mclenachan, Patricia A. P0254,P0420
Mcloughlin, Stephen
Mcmahon, Michelle
Mcsteen, Paula
Meagher, David
P0887
SYM089,P0882
SYM152
P0888
Meagher, Richard
Medina Bujalance, R.
Medina, T
Medrano, Mónica
Meerow, Alan
Meinke, Svenja
Meira, Cleber J.
Meira, Renata
SYM061
P0051
SYM011
SYM015,P0116
SYM106B,SYM109,
SYM145,
P0889,P0890
P0298,P0299 Meghwal, Raju R.
Mehrotra, Rakesh
Mei, Wenbin
Meicenheimer, Roger
Meier, Stuart
Meijaard, Erik
Meimberg, Harald
SYM101
SYM119
P0660,P0700
SYM153
SYM110
P0961
SYM104
P0661
SYM089B,
P0117,P0661
P0118 Meira-Neto, Joao
Melia, Nina
Mello, M.A.R
Melnik, Hadas
Meloni, M.
Melosik, Iwona
Melzer, Siegbert
Memizowa, M.V.
Memmott, P
Ménard, Rozenn
Méndez, Marcos
P0122
P0107,P0131
SYM066
SYM127
P0119
SYM069B
SYM090
SYM104B
SYM123
SYM164
Menendez Martinez, M. P0050
Menezes, Nanuza Luiza SYM069B
Meng, Ying
Menz, Myles
Mercier, Helenice
Merckx, Vincent
SYM120, P0891
P0120
SYM045,P0494,
P0520,P0521,
P0601,P0602
SYM124
Author
Meric, Ciler
Merican, Faradina
Merritt, David
Messina, Tainan
Mestriner, M.A.
Metzgar, Jordan
Meudt, Heidi
Meyer, Janaina
Meyer, Rachel
Meyer-Berthaud,
Brigitte
Meyerowitz, Elliot
Meyers, Emma
Michael, Todd
Michalak, Ingo
Michelangeli, Fabian
Micheneau, Claire
Midgley, Guy
Míguez, Fátima
Mihasan, Marius
Miho, Nakahara-
Tsubota
Mii, Masahiro
Mikami, Tetsuo
Mikatadze-Pantsulaia,
Tsira
Milani, Juliana Foresti
Milani, Pascale
Mildenhall, Dallas C
Mildenhall, Dallas M
Miles, Majesta
Mill, Robert
Millar, A.A
Millar, Harvey
Millar, Katherine D.L.
Millar, Tony
Miller, Chuck
Miller, Eliot
Miller, Jill
Miller, Joeseph
Miller, Rebecca
Miller, Renee
Milne, Ricky
Milner, Melita
Milroy, Stephen P
Minami, Yoshiko
Reference
P0354, P0356, P0359
SYM026
SYM167, P0102
P0081,P0082,P0083
P0065
SYM119,P0893,P08
94
SYM083
P0300
SYM107
P0688
SYM067
SYM153, P0592
SYM162
SYM004,P1011
P0895
SYM057
KNS05
P0481
P0585
P0829
P0391,P0392
P0397
P0121,P0122
P0705
SYM065
SYM101
P0729
P1060
SYM119
SYM060A,SYM060
SYM156
SYM065
SYM060A,SYM060
SYM118, SYM134
SYM099B,SYM104
A,SYM104
SYM102B,SYM107
SYM078,SYM089,
P0955,P0956
SYM016,SYM035,
SYM156,P0230,
P0263,P0532
SYM097
SYM162
SYM104
SYM034
P0523, P0566
715

Miwa, Eitaro
Miyachi, Shigetoh
Miyagi, Atsuko
Miyamoto, Junko
Miyauchi, Hiroshi
Miyaura, Tomiyasu
Miyazawa, Yutaka
Mizrachi, Eshchar
Mizuno, Shuhei
Mizuno, Takayuki
Mizuta, Yukiko
Mizutani, M.
Mo, Peng-Qiao
Moat, Justin
Moço, Maria Cecília
Modarres Sanavy,
Seyed Ali Mohammad
Moein, Fatemeh
Mogoei, Roxana
Moguel-Ordoñez,
Eduardo
Mohamed, Abeer
Mohamed, Haji
Mohammadkhani,
Nayer
Moharrek, Farideh
Mohd-Salleh, Faezah
Mohebi, Javad
Mohney, Brian
Mohr, B.A.R
Moieni, Ahmad
Moir-Barnetson, L
Author
Ming, Ray
Mioto, P.T.
Miranda, Evelyn
Reference
SYM096
SYM045,P0494
P0227
Mirre, Virginia P0825
Mirtadzadini, Mansour P0842
Mishler, B. D.
Misra, Renu
SYM098,SYM158
P0010
Misra, S
Mitchell, John D.
Mitchell, Patrick
Mitchell, Randall J.
Mitsui, Toshiaki
Mitter, Neena
Miura, Eriko
Miura, Yuka
SYM036
SYM115
SYM014
SYM015
P0438
SYM060B
SYM069B
P0606
P0535
SYM026
P0329
P0897
P0838
P0840
P0522,P0575
SYM037
SYM145
P0600
P0523
P0075
P0826
SYM102A,SYM102
P0658
P0517
SYM130
SYM031
P0057
SYM107
P1000
P0237
P0843
SYM070
SYM009
SYM051
SYM006,SYM106B
P0316
P0965
716
Moradshahi, Ali
Moraes, Ana Paula
Moraes, Miguel
Moraes, Regina Maria
Morales, Matias
Moran, Gavin
Moran, Robbin
Morawetz, J. J.
Morden, Clifford
Moreno Gutierrez,
Elizabeth
Moreno Saiz, Juan
Carlos
Moreno, Ana Rodrigo
Morgante, Patrícia
Gleydes
Mori, G. M.
Mori, S. A.
Moriguchi, Ryo
Morin, Nancy
Morinaga, Shin-Ichi
Morita, R
Author
Moiseeva, Maria
Reference
SYM006
Mojab, Faraz SYM177
Molano-Flores, Brenda P0125,P0126
Moldowan, J. Michael
Molino, Jean-François
SYM006
SYM088B
Møllel, Birger Lindberg SYM053
Moller, Michael P0059, P0797
Molnar V., Attila
Mols, Johan
Momohara, Arata
Monaghan, Josie
Monda, K.
Monda, Keina
Mondal (Parui),
Sanjukta
Mondal, Amal Kumar
SYM105B
P1022
P1028
P0420
P0524
P0524
SYM001,P0031,P01
27, P0899
SYM001,SYM114,
P0031,P0127,
P0775,P0784,P0881,
P0899
SYM117 Mondragón-Palomino,
Mariana
Monks, Leonie
Monro, Anna
Monteiro, Silvana H.
Montiel, Olga Martha
Moolhuijzen, P
Moore, Anthony
Moore, Jason
SYM009
P1046
P0913
SYM011
SYM160, SYM161
SYM156
SYM023
P0584
SYM105
P0081,P0082,P0083
P0486
P0381
SYM037
P0931
SYM081
SYM108
SYM005
SYM167
SYM030
P0404
P0404
P1002
SYM046
SYM172
P0128
P0239,P0272

Author
Morita, Ryouhei
Moriwaki, Teppei
Moriyasu, Miyuki
Morley, Robert
Moro, Marcelo Freire
Morokawa, Rosemeri
Morris, J
Morris, Paul
Morris, Robert
Moschini-Carlos, V.
Moses, Alan
Moss, Tiffanie
Motallebi, Parastoo
Moteetee, Annah
Motley, Timothy
Motoyama, Ritsuko
Mott, Ellie
Moulia, Bruno
Moura, Tom Adnet
P0455
SYM063
SYM067
P0153
Mouysset, Elise
Moyle, Leonie
Mukai, Seiji
Mukasa Mugerwa,
Tendo Thomas
Mukhtar, Irum
P0740
KNS02
Mozaffarian, Valiollah SYM177
Muasya, Muthama SYM085,SYM091,S
YM129
Muellner, Alexandra N. SYM072,SYM115
Muenster, T. SYM042
P0861
P0129
Muler, Ana Luíza
Mullany, Kate
Muller, Kai
Mummenhoff, Klaus
P0304
P0130,P0192
SYM022
Mungall, Chris
Munzinger, Jerome
Mur, Luis A. J.
Murai, Yoshinori
Murakami, Noriaki
SYM038B
SYM097,P0403
SYM048
SYM108,SYM115,S
YM168,
P0798,P0947,P0953
SYM010
P0605
Murata, Hiroko
Murata, Jin
P0983,P1001,P1026,
P1029
P0357
P0357, P0835,
P0840, P1063
P0406 Muratova, Elena
Murchie, Erik
Murphy, Daniel
SYM028
SYM078,
SYM088B,
SYM089B
Reference
P0303
P0522
P0566
SYM110
P0901
SYM072
SYM119
SYM134,SYM137
SYM137,P0712
P0139
SYM096
SYM123
P0405
SYM129
SYM108
Reference
P0094
SYM083, SYM127
P0902
P0903
P0903
SYM091
SYM177
P0629
SYM005
PL06, SYM037,
P0423
P0904
SYM055
SYM091
P0256
P0131, P0132, P0133
SYM099
SYM116A, SYM119
SYM110
P0794
P0361, P0451, P0455
P0128
SYM061
P0777
P0292
P0134
P0134, P0666
P0794
P0533
P0266, P0529, P0530
P0135
P0861
P0560
P0905
P0645
SYM034
SYM109
SYM052
P0628
P1073
P0607
P0254
P0276
SYM067
P0455
Author
Murray, Brad
Murray, Brian
Murtazaliev, Ramazan
Muscat, K.M.
Muscat, Karen
Musili, Paul. M.
Muthusamy,
Karthikeyan
Muto, Yukari
Mwebaze, Paul
Myburg, Alexander A
Naiki, Akiyo
Naito, Chiharu.
Najafi, Farzad
Nakagawa, Sayaka
Nakahara-Tsubota,
Miho
Nakahira, Yoichi
Nakaji, Masayoshi
Nakajima, Keiji
Nakamura, Hirofumi
Nakamura, Kyoko
Nakanishi, Tomoko
Nakano, Akihiko
Nakaoka, Masahiro
Nakata, Masashi
Nakatani, Makoto
Nakatsuka, Takashi
Nakayama, N
Nakazono, Mikio
Myles, Benjamin
Mylne, Joshua
Naczi, Robert
Nadaf, Altafhusain
Nadia, Tarcila
Nadot, Sophie
Nagalingum, Nathalie
Nagamasu, Hidetoshi
Nagamasu, Hidetoshi
Nagamura, Yoshiaki
Nagano, Atsushi J.
Nagawa, Shingo
Nagy, Edina
Nahuz, Marcio Augusto
Rabelo
Naidoo, Gonasageran
Naidoo, Yougasphree
717

Author
Nakkuntod, Maliwan
Namiki, Nobukazu
Nampy, Santhosh
Reference
P0664
P0361, P0451
P0907
Nan, Wei-Jiang
Nandan, Suresh
Naquin, D
Narayanasamy,
Veerappan
Narbona, E.
Narimatsu, Shintaro
P0207
P0136, P0908
SYM157
SYM163
P0754
P0507
P0327 Narongajavana,
Dr.Payattipol
Narsai, Reena
Naseem, Anwar
Natale, Christopher A
Natsuaki, Tomohide
Nauheimer, Lars
Navajas-Pérez, Rafael
Nazareno, A.G.
Nazemi, Ghasemali
Nazemi, Sahba
Neale, Alan
P0557
SYM096
SYM062
P0274
SYM106B
P0407, P0408
P0063, P0064, P0065
P0305
P0305
SYM024, SYM053,
SYM054, P0382,
P0465
SYM095 Neale, Richard
Nebel, Martin
Nee, Michael
SYM027, P0400
P0765
Negi, J. P0524
Negri Bernardino, Paulo P0138
Negri, Giuseppina
Negritto, Maria A.
Neinhuis, Christoph
Nejadsattari, T
Nelissen, Hilde
Nelson, David C
Nelson, Gareth
Nelson, Wendy
Nemchinov, Lev G.
Nemomissa, Sileshi
Nepi, Massimo
P0297, P0546
SYM100
SYM080, SYM151,
P0804
P0529, P0530
SYM071
SYM153
SYM104B, SYM121
SYM075
Nepokroeff, M.
Neri, Andreza
Nery Cardoso, Ivan
Ness, Rob
Neubig, Kurt
Neuffer, B
Neumann, P.
P0231
P0924
P0665, P0683,
SYM058
SYM074
P0227, P0232
P0138
SYM102
SYM117
SYM082
SYM038B
718
Author
Nevill, Paul
Nevo, Aviatar
Nevo, Reinat
Newbigin, Ed
Newbigin, Edward
Newell, Christine
Newman, K
Newsham, Kevin
Newton, Angela
Ng, Sophia
Ngan, Bui Thi
Ngo, Q
Nguyen, Chung Huy
Nian, Liu
Nic Lughada, Eimear
Nicholas, Ashley
Nicholas, Richard A
Nichols, Phil
Nichols, R. A.
Nichols, Richard
Nicholson, N
Nickerl, Julia
Nicolas, Antoine
Nicolini, Eric
Nicolle, Dean
Nicolson, N
Nicolson, Sue
Nicotra, Adrienne
Nie, Zelong
Nieder, Jürgen
Nielsen, Nicola
Nieto Feliner, Gonzalo
Nihei, S. S.
Niiyam, K.
Niki, Teruo
Nikkhah, M
Nilova, M
Nilus, Reuben
Nimbalkar, Mansing
Nina, V
Ninkovic, Velemir
Niño-Alcocer, M.
Niroula, Raj Kumar
Nishihara, K
P0172
P0631, P0667
P0405
SYM079A
SYM110
P0032
SYM011
SYM010
P1067
P0409
P0283
SYM162
SYM038B
SYM107
SYM118
SYM165
SYM079B, P0895
P0688
P0423
SYM134
SYM058
SYM014, SYM020,
SYM022, SYM126,
P0024
SYM120, P0891
P0095
SYM064
P0910
P0957
Reference
SYM121
P0326
SYM068
SYM126
P0077
SYM063
P0483
SYM023
SYM122
SYM063
P0440
SYM060
SYM171
P0306
KNS10, SYM011,
SYM118
P0666, P0738, P0803
SYM157

Novoa, Patricio
Novoplansky, Ariel
Nowicki, Christoph
Nowogrodzki, Anna
Nowruzi, Bahareh
Nozaki, Mamoru
Nunes, Claudinéia
Ferreira
Nunez Martin, C.
Nunn, Mike
Nunotani, Tomoo
Nuraliev, Maxim
Nurtjahya, Eddy
Nussaume, Laurent
Nussbaumer, Y
Nyffeler, Reto
O’Donnell, N
O’reilly-Wapstra,
Julianne
O’Rourke, T
Author
Nishihara, Masahiro
Nishimura, Mikio
Reference
P0276
SYM070, P0303,
P0561
P0139, P0140 Nishimura, Paula Yuri
Nishimura, Takeshi
Nishino, Takako
P0575
P0911
Nishinosono, Tatsunori P0535
Nishiuchi, M P0911
Nitta, Joel
Niu, Yang
Nixon, Kevin C.
Nkang, Ani
SYM095
SYM172, P0218
SYM101, SYM121
P0141
Noben, Sarah
Noblick, Larry
Nocentini, Daniele
Nock, Catherine
Nogami, Tatsuya
Noguchi, Hideyuki
Nogué, Fabien
Nomura, Hironori
SYM122
SYM109
P0665
SYM088
P0391
P0142
SYM153
P0560
Nomura, Takahito
Nong, Bao-Xuan
Nonomura, Maiko
North, Clare
Norton, Robert
Novaes, Renan Milagres
Lages
Novák, Petr
P0579
SYM114
SYM042
SYM091
P0176, P0194, P0310
P0875
SYM038B,
SYM107, SYM157
SYM017, P0067
SYM020
SYM103
SYM107
P0529, P0530
P0531
P0366
P0049
KNS07
P1028
SYM169
SYM110
SYM052
P0483
P0695
P0263
SYM149, P0466
SYM104B
P0554
P0684
SYM046
P0075
SYM071
P0669
P0960
P0159
SYM046
SYM039
P0533
Reference
P0889
SYM127
P0447, P0591
P0895
P0144
SYM061
SYM024, SYM053,
P0382
P1064
P0483
SYM020
P0552
SYM156
P0145
P0532
P0146, P0912
P0275
SYM105B
P1065, P1000
SYM041
P0508
P0392
SYM071
P0283
P0840, P1063
SYM095, SYM116B
P0405
P0534, P0855
P0561
P0174
SYM069B
P0307
P0757
P0100, P0130,
P0147, P0167, P0192
Author
Oberli, Andreas
O'brien, Eleanor K.
O'brien, Philip A.
Ocampo, Gilberto
Ocampo, Mery
Oda, Yoshihisa
O'donnell, Natalie
Oelkers, Karsten
Offler, C
Offord, Cathy
Ogasawara, M.
Ogata, Takafumi
Ogawa, Shigeyuki
O'gorman, Kiara
Ogundipe, Oluwatoyin
Ogura, Kenji
Ogura-Tsujita, Y.
Oguri, Emiko
Oh, Dong-Ha
Oh, Soon Ja
Ohashi, Hiroaki
Ohbayashi, Iwai
Ohbu, S
Ohi-Toma, Tetsuo
Ohlsen, Daniel
Ohmori, Tomoko
Ohniwa, Ryosuke L.
Ohsawa, Yukiko
Ohtake, K.
Ohtani, Misato
Ohtsuka, Kurataka
Oikarinen, Emilia
Oja, Tatjana
Oka, Takuji
Okada, Takashi
Okamoto-Nakazato,
Akane
Okhovvat, M
Okitsu, Susumu
Okuyama, Yudai
Olalde, Gabriel
Oliveira Costa, Gisele
Oliveira, Eduardo
Oliveira, Mariana C.
Oliveira, Rafael
719

Author
Oliveira, Reyjane
Patricia
Oliveira, Sabrina
Oliver, M
Olmstead, Richard
Olowokudeje, J.D
Olsen, Kenneth
Olson, Mark
Reference
P0913
P0020, P0148
SYM046, SYM116
SYM077A
P0912
P0808
SYM114
Olteanu, Zenovia P0585
Omara-Achong, Theresa P0141
Omidbaigi, Reza
Ono, Kota
Oohara, T.
Oostermeijer, Gerard
P0316, P0543
P0535
P0411
PL03, SYM127
Ori, Naomi
Orlovich, D.A.
Ortega, R
Ortiz, M.A.
SYM066
SYM172, P0904
P0144
SYM126
Ortiz-Barrientos, Daniel KNS02
Ortiz-Perez, Mario P0057
Ortuñez, Emma
Osborne, Colin
Ose, Naoki
P0796
SYM050, P0516,
P1066
P0535
Oshino, Takeshi
Oskolski, Alexei
Ostria, Enrique
Ostroumova, Tatiana
P0273
SYM079A,
SYM079B, SYM169
P0024
P0706
Osunkoya, Olusegun O P0614
Otani, Ayumi P0828
Otoni, Wagner
Otsuka, Koichi
Otsuka, Kurataka
Ouédraogo, Amadé
P0020
P0392
P0669
P1037
Ouédraogo, Oumarou
Owens, Simon
Oxelman, B.
Oyama, Eriko
Oyama, K.
Oybak Dönmez, Emel
Ozias-Akins, Peggy
Ozturk, Fatmanur
P1037
SYM097
SYM074
P0411
P0886
P1033
SYM039, SYM162
P0351
P. Lowry Ii, Porter
P¨¦Rez, Fernanda
P1040, P1041
P0821
Pabon, Natalia SYM076B,
SYM099B
Pacheco-Figueroa, Coral P0057
720
Parducci, Laura
Paredez, Alex
Parish, Roger
Parisod, C.
Pariyar, Shyam
Park, Kyong-Cheul
Park, Robert
Park, Soo-Chul
Parks, M
Parks, Matthew
Parnmen, Sittiporn
Parsons, Mark
Pasqual, Moacir
Passalacqua, Nicodemo
Giuseppe
Pastore, J.F.B.
Pastova, Ladislava
Paszko, Beata
Patil, Aravind
Paton, A
Author
Pacini, Ettore
Padgurschi, Maíra
Padmavathy, S
Padovan, Amanda
Padurariu, Claudia
Paige, J
Paige, Jay
Reference
SYM058, P0665
P0150
P0322
SYM121
P0585
SYM118, SYM134
SYM134
Paik, Weonki
Pailler, Thierry
Paiva, E.A.S.
Pakmehr, Yazdan
P0851
SYM057
P0040, P1006
P0536
Palacios-Rios, Monica
Palma-Silva, Clarisse
Palmer, Sarah
Palmgren, Michael G.
Palta, Jairo
Pandith, Javid
Pang, Chun-Chiu
Pannell, John
Pansarin, Emerson R.
Panstruga, Ralph
Pant, Bikram Datt
Papong, Khwanruan
P1089
SYM002, SYM105,
P0926
SYM013
P0231
SYM034, P0307
P0308
P0914
SYM102B, P0383
SYM069A
SYM064
SYM052
P0915
Pappas Jr, Georgios A PL06
Paranhos Filho, Antonio P0012
SYM013
PL10
SYM160
SYM096
SYM049B
P0395, P0505
KNS08
P0444, P0445, P0514
SYM119
SYM007
SYM093
P0035
P0364, P0367
SYM139
P0816
P0880
SYM100
SYM018
SYM011, SYM118,
SYM134, P0816

Author
Patrick, Sagmeister
Reference
P0074
Paul, Nigel SYM023
Paulauskas, Algimantas P0226, P0449, P0919
Pauley, Markus
Paul-Victor, Cloé
Paun, Ovidiu
Paungfoo-Lonhienne,
Chanyarat
Pauw, Anton
Pauwels, Maxime
Payne, John
Paz Ares, Xavier
Pazoki, Abbas
Peakall, Rod
Pearse, Stuart J.
Pedersen, Ole
Peery, Rhiannon
Peharec, Petra
Peirson, Jess
Pell, Susan K.
Pellino, M.
Pemberton, Robert
Penailillo, Patricio
Penev, Lyubomir
Peng, Bin
Peng, Ching-I
Peng, Feng
Peng, Tao
Pengelly, Jasper
Penin, Aleksey
SYM046
SYM151
SYM105B, SYM123
SYM031
SYM058
SYM171
SYM110
SYM052
P0458, P0539
SYM057, SYM073,
P0120, P0154,
P0920, P0921, P1014
SYM052
SYM012, P0569
SYM119, P0412
P0542
SYM081
SYM115
SYM039
SYM142
P0067
SYM136
P0815
SYM008, P0769
P0337
P1035
SYM028
SYM169, P1047
Penneys, Darin
Pennington, Toby
Penny, David
Peregrym, Mykyta
P0895
SYM077B,
SYM089A
P0254
P0922
Pereira, Aianã P0227
Pereira, Antonio Batista P0153
Pereira, Caio G.
Pereira, Olinto
Pereira, Paula Natália
P0147
P0148
SYM045, P0494,
P0601, P0602
P0065 Pereira, R.A.S.
Pérez García, N.
Pérez López, David
Perez, Fernanda
Pérez, I
Pérez-Frances, J.F.
SYM167
P0407
P0821
SYM011
P0611
Author
Pérez-Olea, Pedro
Pergl, Jan
Perkins, Andrew J.
Permana, Andrea
Perret, Mathieu
Perrie, Leon
Pertl, Margareta
Pesenti, Antonia
Peter, Bernhardt
Peters, Jonathan
Peterson, Paul
Petit, Rémy J.
Petit, Sophie ('Topa')
Petroli, Cesar D
Petrzelova, Irena
Pettersson, Jan
Pettolino, Filomena
SYM010
P0589
Pevalek-Kozlina,
Branka
P0542
Pfannschmidt, Thomas SYM063
Pfeil, Bernard
Phadvibulya, Valailak
Pharmawati, Made
Phillips, N.G.
SYM074
P0353
SYM052
SYM049B
Phillips, Ryan
Phillipson, Peter B.
Phoenix, Gareth
Phua, Su Yin
Phutthai, Thamarat
Phylogeny Group (Spg),
Sapindales
Pickup, Melinda
Pignal, M.
Pimenov, Michael
Pinheiro Da Costa,
Denise
Pinheiro, Fabio
P0154, P0921
SYM118, P0814
SYM022
SYM054
P0327
SYM115
SYM102B, SYM127
SYM005, SYM128
SYM079A, P0924,
P0941
SYM118
Pinkard, E.A.
Pinto, José Roberto
Rodrigues
Pioch, Karl
Pirani, José R.
Pires, J Chris
Pirie, Michael D.
Pither, Jason
Pitts, D
Piwczyski, Marcin
SYM002, SYM105,
P0926
P0466
P0008
P0312
SYM069A, P0746
SYM096,
SYM117B, SYM157
SYM084, SYM103
SYM144
P0164
SYM079B
Reference
P0053
SYM018
SYM057
SYM110
SYM077A
SYM095
P0848
P0656
PU03
SYM060
SYM100, P0101
SYM088B
SYM057, SYM165
P0423
P1088
721

Author
Plackova, Ivana
Planchuelo, Ana
Pleines, Thekla
Pleurocarps
Evolutionary Research
Group, Perg
Plume, Olofron
Reference
P0113
SYM003
P0928
SYM122
Plunkett, Gregory M
Pocius, Olidan
Pócs, Tamás
Poethig, Scott
Pogson, Barry
SYM120
SYM079B, P0929
P0009
SYM118
SYM066
Poiré, Richard
Pokorny Montero,
Cristina Isabel
Pokorny, Lisa
SYM054,
SYM063,SYM068,
P0620
P0540
SYM122
Poldy, Jacqueline
Poli, L. P.
Pollard, G
Pollnac, Fred
Pompêo, M.L.M.
Ponce De Leon, Marcia
S
Poncy, Odile
SYM122
SYM073, P0920
P0689
P0164
P0014
P0139, P0140
SYM139
Poopath, Manop
Pooyan, Paria
Popic, Tony
Popp, Magnus
Poppinga, Simon
Porembski, Stefan
SYM005
P0327
P0799
P0233, P0234
P0825
SYM151
SYM002, P0782
Pornsiripongse,
Saowapa
SYM177
Pornsiriwong, Wannarat SYM054, SYM063,
P0620
Porter-Morgan, H P1045
Posch, Sabine SYM034, P0176,
P0194, P0310
Poschenrieder, Charlotte SYM024, P0155
Possingham, Hugh KNS08
Potgieter, Christina
Potter, Daniel
Potts, Brad M.
P0930
SYM150
SYM121, SYM149,
P0466
SYM110 Poulsen, Axel D
Pourmirzaii, Ahmad
Powell, Jeff R
Powles, Steve
Prado, Jefferson
SYM009
SYM016
P0102
P0931, P0932
Prasad, S SYM036
Prathanturarug, Sompop SYM177
722
Author
Pratt, Renae
Pratt, Steven
Preece, Mike
Preiss, Katja
Prenner, Gerhard
Prescott, Matthew
Pressel, Silvia
Preston, Christofer
Preston, Jill
Price, Charles
Price, Michelle Judith
Price, Sarah
Prinzing, A.
Proença, C.E.B.
Prokhorov, Alexey
Reference
P0933
P0798
P1075
SYM124
SYM076B
SYM078
SYM087
P0960
SYM099A
P0541
SYM005
P0223
P0072
SYM131
SYM082
Proust, Hélène
Pruesapan, Kanchana
Pruitt, N
Prusinkiewicz,
Przemyslaw
Prychid, Chrissie
Pryer, Kathleen
SYM153
P0989
SYM116
SYM067, PL04
SYM091
SYM086, SYM088B
Puente-Lelièvre,
Caroline
SYM104B
Puripunyavanich, Vichai P0352, P0353
Pyle, Richard P0078
Pyšek, Petr
Pyvovarenko, Tatiana
Qaiser, M
Qi, Zehchen
Qiu, Yin-Long
Qu, Dongyu
Qu, Lina
Quail, Peter
Quandt, Dietmar
Queiroz, L.P.
Quick, Paul
Quilliet, Catherine
Quinn, C.
R. Claßen-Bockhoff,
Rabaey, D.
Radic Brkanac, Sandra
Radovski, Monica
Rae, Anne
Raes, Niels
SYM018
P0287, P0380
P0221
P0793
SYM155
SYM107
P1009
PL07
SYM027,
SYM038B,
SYM086, SYM122,
P0400
P0802
SYM028
SYM151
SYM104B
SYM076
SYM049
P0542
P0098
P0312
SYM110

Author
Rafalski, Antoni
Raggi, Lorenzo
Raghothama,
Kashchandra G
Ragni, Laura
Raguso, Robert
Reference
P0589
SYM162
SYM052
SYM153
P0224
Rai, H. S.
Rai, Hardeep
SYM049B,
SYM119, Sym124
SYM119
Rajanikanth, Annamraju P0934
Rajeshirke, Sneh
Rakotomalala, Jean-
Jacques
Ralph, Paula
Ramachandran,
Veerambakkam
Srinivasan
Ramak*, Parvin
P1097
SYM066
P0377
P0200
P0284
Ramamurthy, Kailash B P1100
Ramaraj, Jayakumararaj SYM177
Rameau, Catherine
Rammul, Kadi
SYM153, p0594
P0159
SYM095 Ramp Neale, Jennifer
M.
Ramsden, Lawrence
Ramsey, Mike
Ramula, Satu
Ranaivo, Jaona
P0156
SYM015
SYM014
P1040,p1041
Randrianasolo,
Razafiharimina Marie
Agnès
Rane, Gauri
P0967
P0157, p1092
Rangappa, Kavyashree P0313, p0314
Rangsiruji, Achariya
Rani, Arti
Ranker, Tom A.
Rao, G-Y
Rao, N.
Raouf Fard, Fatemeh
Rapini, Alessandro
Rasmus, Sirpa
Ratcliffe, R. George
Rather, M.A
Rather, M.A.
Rathi, Neelima
Rattey, A
Raubeson, L
Raubeson, Linda A.
Rauchová, Jana
Raven, John
SYM093
SYM159
SYM027, SYM095,
P0932
P0870
SYM071
P0316, P0543
SYM072
SYM022
SYM156
SYM163
P0778
P0252
SYM160
SYM119
SYM119, P0412
SYM105, SYM105A
SYM026
Author
Raven, Peter
Raven, Peter H.
Ravensdale, Michael
Reference
KNS09, SYM118
KNS09, PU02
SYM055
P0158 Ravi Raja Selvi, Asir
Selin Kumar
Ravishankar, G.A.
Rawat, Anil Singh
Rawls, Michael
Rawsthorne, J
Razanajatovo, Mialy
Harindra
Razavizadeh, Roya
P0565
SYM163
SYM056
SYM001
P0967
Rebetzke, G J
Reca, Barbara
Reche, María
Redden, Bob
Reddy, Peddi Gopal
Reese, Zulkifly
Rei Souza, Mld
Reich, Ziv
Reichman, Suzie
Reid, A.
Reid, James
Reid, JB
Reier, Ülle
Reinheimer, Renata
Reis, M.S.
Reis, Talita
Reis, Talita Soares
Reiter, Noushka
P0536, PO603
SYM036
SYM059
P0407
P0291
P0317, PO318
SYM026
P0732
SYM068
SYM023
SYM127
P0643
P0485
P0159
SYM076B
P0063, P0064
P0161
P0161, P0162, P0163
P0164, P0935
Remizowa, Margarita V. SYM090
Ren, Hai SYM009
Ren, Haiyun
Renaut, Sebastien
Rengel, Zed
Rennebarth, Thorsten
SYM061
KNS02
P0231
SYM018
Rennenberg, Heinz
Renner, Matt A. M.
Renner, Susanne S
Renny-Byfield, Simon
Rentsch, D.
Renzaglia, Karen
Requerey, Rocío
Resende, L.C.
Retha, Meier
Rew, Lisa
SYM030, SYM051
SYM118, P0936
SYM106B,
SYM106B, SYM136
SYM038B,
SYM107, SYM157
SYM031
SYM087, SYM155
SYM015
P0104
PU03
P0014
723

Author
Rexhepi, Jashar
Rey, Herve
Reyes, M.E.C
Reference
SYM136
SYM067,P0688
SYM060
Reynders, Marc
Rezende, Fernanda
Mendes
Ribas-Carbo, Miquel
Ribeiro, Ana Paula
Ribeiro, Larissa
Ribeiro, Patrícia Luz
SYM091
P0486
SYM156
P0366
Ribeiro, R.A. P0104
Ribes De Lima, Letícia P0297, P0872
Ricardo Sodre Cardoso,
Sergio
Ricca, Mariana
Rice, Kevin
P0020
SYM072
SYM077B
SYM098
SYM025, P0961
Richards, Christina
Richards, K
Richards, Richard A
Richardson, James E.
Richaud, Frédérique
Richter, Katherine
Rico Arce, Maria De
Lourdes
Rico-Gray, Victor
SYM020
SYM118
KNS06, SYM036
P1074
SYM162
SYM156
SYM005, P0748
Rieseberg, L.H.
Rieseberg, Loren
Righi, Adne
Riina, Ricarda
Rillig, Matthisas C
Rio, M.C.S.
Ripin, Azizi
Ripley, Brad
Rival, Alain
Rivas Acuña, Ma.
Guadalupe
Rivas Arancibia,
Sombra Patricia
Rivers, Malin
SYM150
SYM090, SYM120
KNS02, SYM120
P0546
SYM004, SYM081
SYM016
P0680
P0172
SYM050
SYM162
P0937
P0166
Roalson, Eric
Robbrecht, E.
Robert, Jeyachandran
Roberts, David G.
Robertson, Alastair
Robertson, Masumi
Robinson, Nicole
Robinson, Sharon A.
Robischon, Marcel
KNS10
SYM085, SYM091
SYM137
P0547
SYM127
SYM001
SYM060
SYM030, SYM031
SYM022, SYM023,
SYM156 P0091,
P0187,P0230
SYM069B
724
Author
Robles, Francisca
Rocha-Jorge, Renato
Rodrigues, Jacqueline
Gomes
Rodrigues, M.A.
Rodrigues, M.E.F.
Rodríguez, J.A.
Rodríguez, Nicolás
Rodriguez-Avalos, Jose
Alberto
Rodriguez-Medina1,
Caren
Roeder, Adrienne
Roesch, Luiz Fernando
Würding
Rogers, Hilary
Rohlf, F.James
Rokshar, Daniel S
Reference
P0407, P0408
P0756
P0875
SYM045
P0140
P0144, P0184,
P0610, P0611
P0779
P1067, P1068
SYM052
SYM067
P0153
SYM070
SYM139
PL06
Roleda, Michael
Rolland, Norbert
Romanov, Mikhail
Romaschenko,
Konstantin
Ron, E.
Roncal, Julissa
Ronquist, Fredrik
Ronse, Anne
Rønsted, Nina
SYM026
SYM068
P0672
SYM100
P0609
SYM077B
SYM103
SYM167
SYM117,
SYM117A, SYM145
SYM110, SYM137 Roos, Marco C.
Rose, Evan
Rose, Pamela
Rose, Ray
Rosell, Julieta A.
Rosique-Gil, E.
Ross, JJ
P0977
SYM013
SYM061
SYM114
P0750
P0485
Rossetto, Maurizio D.R SYM020, SYM092,
SYM104, P0556,
Rossi, Graziano
P1062,
P0002
Rossouw, A
Rotchanapreeda, T.
Rothfels, Carl
Roth-Nebelsick, Anita
Rothwell, Gar
Rouhan, Germinal
SYM079, SYM079A
SYM171
SYM088B
SYM151
SYM007, SYM119
SYM116B, P0931,
P0932, P0814
SYM062 Rounds, Caleb
Rout, Tracy
Routier, AL
Rouzé, Pierre
KNS08
SYM067
PL06

Author
Rowe, Nicholas
Rowntree, Jennifer
Roy, Mélanie
Roycewicz, Peter
Rudall, Paula J.
Ruffatto, Danielle
Ruhsam, Markus
Ruiz Herranz, A.
Ruiz Rejón, Carmelo
Ruiz, Eduardo
Ruohan, Wang
Russell, Anton
Russell, Joanne
Russell, Scott
Ruthrof, Katinka
Ruzvidzo, Oziniel
Ryan, Megan H
Ryan-Colton, Ellen
Rydberg, Anders
Rydin, Catarina
Rye, Barbara
Rymer, Paul D.
Saadati, Nasim
Saarela, Jeffery M.
Saarinen, Timo
Sabatier, Daniel
Sabatier, Sylvie
Sack, Laweren
Sadamitsu, Atsushi
Sadovnikova, E.S.
Saeidi-Sar, Sakineh
Saenz, Gabriel
Sáez, Llorenç
Safari, Masoumeh
Safikhani, Keivan
Sagade, Ashok
Sage, Tammy
Sahoo, R.
Sahuquillo Balbuena,
Elvira
Saida, Yuka
Saito, Fumie
Saito, Susum
Saitoh, Kenji
Reference
SYM151, P0804
SYM149
SYM105, SYM105A
SYM046
SYM076B, SYM090
P0126
SYM119
P0051, P0788
P0407, P0408
P0067
P0674
P1042
SYM059
SYM060,SYM060A
SYM024
SYM153
SYM162
SYM022, P0091
P0222
SYM119, P0222
P1069
SYM020, SYM099
SYM130, P1070
SYM100, P0938,
P0939
SYM022
SYM088B
SYM066, P0687,
P0688
SYM014, SYM056,
SYM116B,
SYM172, P0682
P0940, P0968
SYM090
P0458, P0459,
P0460, P0549, P0550
P0167
P1052
P0487
SYM009
P0414
SYM050, SYM152
P0511
P0924, P0941
SYM040
SYM046
P0667
P1073
Author
Saitoh, Tomoyo
Sajo, Graça
Sakaguchi, Keisuke
Sakai, A.K.
Sakai, Hidemitsu
Sakai, Kaori
Sakai, Yuuki
Reference
P0438
SYM076B
P0604
SYM074
SYM034
P0605
SYM042
Sakakibara, H.
Sakata, Tadashi
Sakio, Hitoshi
Sakkir, Sabitha
Sakuma, Yoh
Salamin, Nicolas
Salatino, A.
Salatino, Antonio
Saleeba, Jenny
Saleh, Said
Salehnia, Ali
Salimpour, Fahimeh
Salinas, Paula
Salles Sampaio, Diana
Salomo, K.
Salomo, Karsten
Saltmarsh, Anna
Salvi, Daniel
Salvo, Gabriele
Samain, Marie
Stephanie
Samanta, Shashikar
P0551, P0606
SYM077,
SYM077A, P1066
P0300, P0546
P0297
P0129
P0319
P0266, P0267
P0568
SYM153
P1099
SYM080, P0804
SYM080
KNS10
SYM068
SYM171
SYM080, P0804
P1024
Samigullin, Tahir P0990, P0991
Samuel, Rosabelle SYM004, SYM168,
P0947, P1042
San'ana, Gustavo César P0365
Sánchez-Perez, Raquel SYM053
Sanderson, M. J.
Sanderson, Michael S.
Sandoval, Estela
Sanmartin, Isabel
SYM089,
SYM089A, P0883
SYM089, SYM150
P0948, P0949
SYM103, P0721
Sano, Satoshi
Sansaloni, Carolina P
Santos Filho, Laf
Santos, Alexandre
Santos, Deborah Yara
A.C.
Santos, Flavio Antonio
Maës
Santos, M.C.J.
P0524
P0273
P0168
P0169
P0560
P0423
P0731, P0732
P0114
P0486
P0162
P0680
725

Author
Santuari, Luca
Sanz, Ernesto
Saporetti-Junior,
Amilcar
Sarafis, Vassilios
Saralamp, Promchit
Reference
SYM153
P0796
P0118
Sarkar, P.
Särkinen, T
Sarmento Cabral,
Juliano
Sasaki, Karen Lucia
Mayumi
Sasaki, Tadamasa
P0675, P0676
SYM177
P0631
SYM077B
SYM144, SYM144A
SYM069B
Sasaki, Taku
Sasidharan, R
Saslis-Lagoudakis, C.
Haris
Saswade, Ramkisan
P0552, P0559
SYM123
SYM012
SYM145, P0952
Satish, M
Sato, Kaori
Sato, Seiichi
Sato, Shusei
Sato, Tamotsu
Sato, Yasushi
Sato, Yutaka
Satoh, H.
Satoh, Shinobu
Sato-Nara, Kumi
P0553
P0518
P0277
P0496
SYM161,SYM162
P0172, P0272
P0496, P0531, P0554
P0361, P0451, P0455
P0559
P0499
P0677
Sattaripour, Mahdiyeh P0568
Saumitou-Laprade,
Pierre
SYM171
Saunders, Richard M. K. SYM086, P0914,
P1022
Sauquet, Herve SYM099
Sautter, G.
Savinov, Ivan
Savolainen, Vincent
Saw, L.G.
Saw, Leng Guan
Sawa, Shinichiro
P0712
SYM076B
SYM145
SYM110
SYM128
SYM042, P0440,
P0442, P0443,
P0446, P0453, P0454
SYM145
SYM058
Sawai, Satoru
Sawhney, V K
Sawtell, W
Scacchi, Emanuele
Scaffidi, Adrian
Scalon, Marina Corrêa
Scanlon, Annette
Scasitelli, Moira
Scataglini, M. Amalia
SYM085
SYM153
SYM153
P0555, P0556
SYM165
KNS02
SYM100
726
Author
Scatena, V.L.
Schaal, Barbara A
Schaefer, Carlos
Schaefer, Didier
Schaefer, Hanno
Scharaschkin, Tanya
Scharf, Alexander R.
Reference
P0802
P0763, P0769
P0118, P0227
SYM153
SYM084
P0614, P0914
P0951
Schatz, George E. P0814
Scheible, Wolf Ruediger SYM052
Scheidegger, Christoph P0182
Scheinvar, Leia P0174
Schenk, H. Jochen
Scheupback, T
Schiestl, Florian
Schire, B D
Schlauch, Karen A
Schlessman, Mark
Schlüeter, Philipp
Schmickl, Roswitha
Schmid, Markus
Schmidt, Alexander R.
Schmidt, Marco
Schmidt, Susanne
SYM049, SYM049A
SYM067
SYM073, SYM169
SYM129
SYM045
SYM079, P0953,
P0954
SYM169
SYM097
KNS03
SYM155
SYM017, P0782,
P1037, P1039
SYM030, SYM031,
SYM104B
SYM170 Schmidt-Lebuhn,
Alexander Nikolai
Schmitt, I
Schmutz, Jeremy
Schnabel, Andrew
Schnable, P S
Schnadelbach,
Alessandra
Schneebeli, Katharina
Schneider, Harald
Schneider, Julio V.
Schnitzler, Jan
Schoen, Dan
Scholz, Fabian G.
Scholz, Henning
Schomaker, Karsten
Schönenberger, Jürg
SYM035
PL06
P0955, P0956
SYM157
P0913
P1098
SYM086, SYM095
P1039
SYM103
SYM096
P0588
P0398
SYM082
Schranz, Eric M
Schreiber, Lukas
Schrey, Aaron
Schrire, Brian
Schuette, Scott
SYM099,
SYM099A,
SYM099B, P0695
SYM096, SYM157
P0587
SYM020
SYM089,
SYM089A, SYM129
SYM048

Author
Schuettpelz, Eric
Schulman, Leif
Schulte, Katharina
P0664
SYM060B
SYM060B, P0620
SYM084
P0404
SYM049
P0557
SYM114
P0286, P0543
SYM064
P0416
SYM151
SYM068
P0606
P0579
SYM034
SYM105A, SYM124
P0678
SYM158
SYM034, SYM035,
P0176
SYM038B
SYM164
SYM149
P0515
SYM059
P1015
SYM056
SYM073
SYM088B
SYM088B
Reference
SYM086, P0931
SYM082
SYM002, SYM004,
SYM017,
SYM077B, SYM103
SYM046
P0854
P0452
P0386
SYM168
SYM004
SYM034
SYM018
P0957
P0957, P1094
Schultz, Carolyn
Schulz, Christian
Schuster, Gadi
Schuster, Stefan
Schuster, Tanja M.
Schütz, Nicole
Schwadorf, K.
Schwartz, Mark W.
Schwarz, E. A.
Schwarz, Elizabeth De
Araujo
Schwarzacher, Trude
Schwarzer, Christian
Schweitzer, Jennifer
Schweizer, P.
Schwerdt, Julian
Scoble, MJ
Scoffoni, Christine
Scopece, Giovanni
Scotti, Ivan
Scotti-Saintagne,
Caroline
Seangruttanachaikul,
Irin
Searle, I.R
Searle, Iain
Sebastian, P.
Sebastião, I.
Secchi, Francesca
Secco, David
Sedio, Brian E
Sefidkon, Fatemeh
Sehgal, Anita
Sehrish, Tina
Seidel, Robin
Seigneurin-Berny,
Daphné
Seike, Kumiko
Sekimoto, Hitoshi
Selmair, P.
Selosse, Marc-Andre
Selvam, A. Brama
Dhayala
Séneca, Ana
Seneweera, Saman
Shahidul, Islam
Shan, Yu
Shani, Eilon
Shao, Guihua
Sharbel, Timothy F
Sharifi, Mozafar
Sharma, Madhu
Sharma, Prabha
Sharma, Reetu
Sharp, Robert
Sharwood, Robert
Shavarda, A
Shavrukov, Yuri
Shaw, Blanka
Shaw, Jonathan
She, Kao-Chih
Shelden, M C
Shen, Bo
Shen, Wanxia
Shen, Wen-Hui
Shen, Yuan
Shepherd, Kelly A
Shepherd, Lara
Shepherd, Mervyn
Sheppard, Andy
Sherman-Broyles, Sue
Shetekauri, Shamil
Sheue, Chiou-Rong
Author
Sennikov, Alexander
Sentíes, Abel
Seppelt, Rodney D.
Septiningsih, Endang
Serizawa, Shunsuke
Serrano Maldonado,
María José
Sessa, Emily
Sethuraman,
Padmavathy
Seto, Yoshiya
Seung, D
Severns, Paul
Sexton, Jason
Sexton, Timothy
Sforça, D.A.
Shabala, Lana
Shabala, Sergey
Reference
P0959, P0960
P0756, P0757
P1000
SYM012
P0392
SYM001
SYM116. SYM116A
P0177, P0322
P0558
SYM160, P0655
SYM127
P0961
SYM037
P0998
P0231
SYM030, SYM070,
P0231
SYM159
P1050
SYM066
SYM038B
SYM039, SYM162
P0284, P0517, P0543
SYM159
P0179
P0252
SYM046
P0452
SYM170
P0326
SYM155
SYM098, SYM122
P0559
SYM049,
SYM049A, SYM061
P0419
SYM060B
SYM123
SYM061
P0209, P0965
P0420
SYM037, SYM088
SYM020
P0421
P0966
P0967, P0985
727

Author
Shi, Xiang
Shibukawa, T
Shiga, T.
Shigenobu, Shuji
Shiina, Takashi
Shimada, H.
Shimada, Motoko
Shimada, Takashi L.
Shimada, Tomoo
Shimamura, Masaki
Shimazu, Toru
Shimizu, Kentaro K.
Shimizu, Noriko
Shimizu-Inatsugi, Rie
Shimma, Yoh-Ichi
Shimoni, Eyal
Shimosato-Asano,
Hiroko
Shin, Young Seop
Shinde, Bajirao
SYM068
SYM064
P0456
P0513
Shindikar, Mahesh P0180, P0204
Shingaki-Wells, Rachel SYM156
Shinji, Handa
Shinohara, Wataru
Shiori, Ishii
Shiota, Hajime
P0829
P0863
P0181
P0679
Shipley, Alan
Shipunov, Alexey
Shirakawa, Makoto
Shirakawa, Y
SYM062
SYM169
P0561
P0283
Shirley, Neil P0287
Shleizer Burko, Sharona SYM066
Shoji, Kazuhiro
Shoji, Tsubasa
P0271
SYM053
Reference
P0236
P0303
P0075
P0453, P0454
P0560, P0684
P0552, P0559
P0507
P0561
SYM061, P0561
P0856, P0940, P0968
P0642
SYM126
P0453
SYM126
SYM046
Shorter, R
Shou, Huixia
Showalter, Allan
Shrestha, Keshab P.
Shrestha, Krishna K.
Shrestha, Uttam Babu
Shulin, Mou
Shuttleworth, Adam
Siddiqi, Tariq Omar
Siddique, A B.M
Siddique, K.H.M.
Siddons, Hayley R.
SYM160
P0557
SYM046
P0097
SYM005, P0182
P1100
P0674
P0803
P0563
SYM041
P0307
P1064
728
Author
Sierra Daza, S.
Sierra Muñoz, José
Carlos
Sikes, Benjamin A
Silva Oliveira, Rafael
Silva Torres, Beatriz
Reference
SYM137
P0184
SYM016
P0023, P0138
SYM003, P0185,
P0806
P0117 Silva, Luzimar
Silva, Paul
Silva, Tania
Silva, W.
Silveira, L. E. D.
Silvera, Katia
Silverio, Adriano
Silvestro, Daniele
P0759
P0971
P0257
P0404
SYM045, P0373
P0657
SYM002, SYM004,
SYM017,
SYM077B,
SYM103, P1011
P0702
P0972
Simão, D.G.
Simão-Bianchini,
Rosangela
Simmonds, J
Simmons, M.P.
Simoes, Andre
Simon, Judy
Simon, Marcelo
SYM046
SYM144B
SYM072, P0616,
P0680, P0852
SYM051
SYM077, SYM089,
SYM150
SYM032 Simon, William
Simpson, David
Singer, Rodrigo
Singh, D.K.
Singh, Geetha
Singh, J
SYM091, P1066,
P1015
SYM073, P0974,
P0975
SYM087
P0308
P0518, P0519
Singh, Kashmir
Singh, Mohan
Singh, Ravi Shankar
Singh, Rohan
Singh, S K
Sinha, Neelima
Siol, Mathieu
Siqueiros-Delgado,
Maria Elena
Sirault, Xavier Rr
Sirisena, Udani M.
Skema, Cynthia
Skog, Laurence E
Slabas, Antoni
SYM159
KNS04, SYM060,
SYM060A
SYM159
P0287
SYM036
SYM069B
SYM102
P0184, P0976, P1068
SYM035, P0540
SYM117
Sivakuruman, Anushen SYM010
Skean, J. Dan P0895
SYM099B,
SYM104A
SYM171
SYM032

Author
Slack, Nancy
Reference
SYM158
Slamet-Loedin, Inez SYM028
Sletten Bjoraa, Charlotte P0825
Slik, J. W. Ferry
Slingsby, Jasper
Alexander
Slotkin, Keith
Small, Ian
SYM110
SYM170
Smets, Erik F.
KNS01
SYM055, SYM063,
SYM068, P0503
SYM049,
SYM069B,
SYM104, SYM115,
SYM124, SYM137
P0832 Šmilauer, P.
Smirnov, Sergey, V.
Smissen, Rob
Smith, Andrew
Smith, Barry
Smith, Colin
Smith, F. Andrew
Smith, Gideon F.
Smith, J.A.C.
Smith, James F.
Smith, Liza
P0597
SYM083, P1071
SYM004, SYM016
SYM048
SYM091
SYM016
SYM117A, SYM118
SYM017
SYM080
SYM023,
SYM060B,P0187
SYM025 Smith, Melissa C
Smith, Neil A
Smith, Oliver
Smith, Penelope M.C
Smith, Richard
Smith, Sally E.
Smith, Steven
Smith, V S
Smith-Huerta, Nancy
Smithson, Ann
Smtih, J Andrew C
SYM060B
SYM013
SYM052
SYM067
SYM016
SYM153, P0084
P1015
P0977
SYM073, P0743
SYM017
SYM092, SYM104B
SYM166
Sniderman, Kale
Snow, Neil
Snowball, Richard
Soares E Silva, Uiara
Catharina
Soares, Bárbara D. F.
Soares, Michellia
Söderström, Lars
Soejima, Akiko
Sokoloff, Dmitry D.
Sola Campoy, Pedro J
Soll, Juergen
Solomon, James
SYM162
SYM072
P0364, P0365
P0118
SYM118, SYM158
P0978
SYM090, SYM169
P0407, P0408
SYM068
P0722
Author
Solomon, Peter
Soltis, Douglas
Reference
KNS08
Soltis, Pamela
Somerville, Chris
Son, Jae-Han
Song, Chi
Song, Fuqiang
Song, Ye-Su
Soni, Shefali
Sookchaloem,
Duangchai
Sorek, Nadav
SYM038, SYM157,
P0377, P0416,
P0890, P0979
SYM038, SYM157,
P0377, P0416,
P0890, P0979
PL10, SYM059
P0395, P0505
SYM038B
SYM082
P0849
P1097
P0327
Soreng, Robert J.
Soriano Santos, Jorge
Sorrell, Brian
Soto Medina, M.
Sousa, G. M.
Sousa, Mario
Southerton, S. G.
Souza, A.P.
Souza, Agostinho
Souza, Alessandra
Souza, Amanda
Souza, C.D.
Souza, M.B.C.
Souza, Rafaeli
Aparecida Vieira
Sovanmoly, Hul
Spalik, Krzysztof
Spangenberg, G.
Španiel, Stanislav
Specht, Chelsea
Speck, Thomas
Sperry, J.S.
Spicer, Robert
Sporck, Margaret J
Spriggs, Andrew
Sprunck, Stefanie
Squeo, Francisco A.
Sramko, Gabor
Sreekanta, S.
PL10
SYM100, P1071
SYM001
SYM012
SYM127
SYM004
P0748
SYM037
P0998
P0118
P0521
P0009
P0704
P0998
P0365, P0367
P0980
SYM079B, P0727
P0393
SYM097
SYM069
SYM151
SYM049
SYM006
P0682
SYM060A
KNS04
SYM165, P0188
SYM105B
P0631
Sridevi, Vaddadi P0565
Srilunchang, Kanok-Orn KNS04
Srinivasan, Bharathi
Srinivasan, Natarajan
P0314
SYM163
729

Author Reference
Staedler, Yannick Marc SYM099B
Stafford, Gary
Staiger, Chris
SYM145
SYM061
Stajsic, Val
Stangoulis, J.
Stanley, Will
Stapf, Maria
SYM018, SYM088A
SYM035
SYM055
P0971
Starr, J.R
Steadman, Kathryn
Steane, Dorothy A
Stech, Michael
SYM085
SYM022
SYM108, P0423
SYM075, SYM098,
SYM122, SYM172
SYM089 Steele, Kelly
Steele, P. Roxanne
Stefan, Dullinger
Stefan, Marius
Stefani, Franck
Stefanovic, Sasa
Steinmann, V.W.
Stenøien, H.K.
Stephen, S.A.
Stephenson, Rus
Sterck, Lieven
Stern, David
Stern, Stephen
Stevens, Albert-Dieter
Stevens, Jason
SYM117B
P0074
P0585
P0982
SYM124
P0868
SYM013
P0062
P0619
PL06
P0452
SYM107
SYM082
P0066, P0102
Stevens, Peter SYM134
Stevenson, Dennis Wm SYM048,
SYM076B,
Stevenson, Pablo
SYM106, SYM109,
SYM119, P0889
SYM164
Stewart, Doug
Stierhof, York
Stilwell, Stephen
Stinchcombe, John
Stirton, C.H.
Stöckel, Marcus
Stockey, Ruth
Stoll, Alexandra
Stone, Graham
Stotler, Raymond
Stpiczyñska,
Ma³Gorzata
Strasberg, Dominique
Strasburg, Jared
Straub, Shannon
P0340
SYM064
SYM023
SYM096
SYM129
SYM124
SYM007
SYM165, P0188
SYM078
SYM155
P0683
SYM027, SYM057,
SYM158
KNS02
SYM038A, P0809
730
Author
Strauss, Sharon
Strellner, Reid S.
Stromski, Jessica
Strong, Mark
Strother, Paul
Struwe, Lena
Stuart, Peter
Stuessy, T. F.
Stuessy, Tod
Stuetzel, Thomas
Su, Tao
Su, Yvonne C. F.
Suárez, E
Subbaiyan, Gopala
Krishnan
Subramanian, Ramya
Suda, Jan
Sugai, Kyoko
Sugai, Michizo
Sugawara, Takashi
Sugimoto, Kazuhiko
SYM177
SYM105A
P0983
P0643
P0905
P0361
Sugimoto, Takanori
Sugiyama, Munetaka
P0560
SYM071, P0531,
P0669
P0424 Sugiyama, Shuichi
Suh, Eun Jung
Suh, Youngbae
P0444, P0445
P0851
Sukhorukov, Alexander P0979
Sukias, James SYM012
Sukkharak, Phiangphak SYM172
Sule, Daniel P0174
Suleiman, Monica
Sultan, Sonia
P1000
SYM020
Sumadijaya, Alex
Sun, B.-Y.
Sun, Hang
Sun, Hao
Sun, Penglin
Sun, Samuel Sai-Ming
Sun, Shan
Sun, Wei
Sun, Weibang
Sun, Xiaoqin
Sundaresan, V
Sundberg, Marshall
Reference
P0961
SYM050
P0378
P0708
SYM169
P0746, P0994
SYM024, SYM053
SYM123, SYM126,
SYM144B, P0067
SYM094B,
SYM123, P0067,
P0854
SYM164
SYM086, P0914,
P1022,
P0611
P0298, P0299
SYM140
SYM144B
SYM120, SYM172,
P0218, P0984
P0596
SYM062
SYM038B
P1043
SYM050
SYM009
P0815
SYM060
SYM140

Author Reference
Sundue, Michael A. SYM027, SYM095,
P0931
Sungsinleart, Kwanpisut P0353
P0240 Sunthornchainaksaeng,
Puangpaka
Super, Laura SYM144
Susanti, R.
Susumu, Okitsu
Sutcliffe, Jill
Sutcliffe, Vanessa
P0897
P0181
SYM166, P1095
P0121
Suto, Takamichi
Suwa, Rempei
Suwastika, I Nengah
Suyama, Akiko
Suyama, Yoshihisa
Suzuki, Go
Suzuki, H.
Svenning, J.-C.
Swain, Steve
Swangpol, Sasivimon
Sweetlove, Lee J.
Syme, Anna
Symmank, L.
Symonds, Vaughan
Sytsma, Kenneth
Tabata, Ryo
Tada, Satoshi
Tajadod, Golnaz
P0642
P0142
P0684
SYM046
P0189
SYM039
P0552
SYM109
SYM152
SYM171
SYM156
SYM088A,
SYM088B, SYM100
P0804
SYM157, P0115,
P0362, P0363, P0416
SYM004, SYM108,
SYM172, P0751,
P0783
SYM042, P0443,
P0453, P0454
P0566
Takabe, Keiji
Takagi, K.
Takahara, Kentaro
Takahashi, Hideyuki
Takahashi, Mitsuo
Takahashi, Takeyoshi
Takahashi, Taku
Takamiya, Masayuki
Takaya, Eliko
Takayama, Koji
Takebayashi, Kohsuke
Takehisa, Hinako
Takeyasu, Kunio
Talavera, M.
Talavera, S.
Talbot, Mark
SYM031, P0568,
P0567
P0699
P0633
P0271
P0276, P0329,
P0522, P0575
P0667
SYM145
P0561
P0076
P0643
SYM144B
P0560
P0303, P0361,
P0451, P0455
P0684
SYM126
SYM126
P1101
Tay, D
Taybi, Tahar
Taylor, Chris G.
Taylor, David
Taylor, Jen
Taylor, Mackenzie
Taylor, Nicolas
Taylor, Sam
Taylor, Sunni
Taylor, Wilson
Teakle, Natasha
Teasdale, Suliana
Teixeira, S.P.
Teixido, Alberto L.
Tejedo, Pablo
Telenius, Anders
Author
Tallavera, Salvador
Talve, Tiina
Tamaki, Takayuki
Reference
P0755
P0159
P0442
Tamaoki, Daisuke
Tamura, Kentaro
P0642
SYM070
Tamura, Minoru N. P1054, P1072
Tan, Ashley Hwei-Ting P0425
Tan, Wee Kee
Tan, Yew-Foon
Tanaka, I
Tanaka, Norio
Tandon, Pramod
Tandon, Rajesh
Tang, Mo Shih
Tanigaki, Fumiaki
P0686
SYM156
P0679
P0392, P1073
SYM080
SYM064
P0985
P0642
Taniguchi, Koji
Tanner, Chris
Tanurdzic, Milos
Tao, Wang
Taprab, Suniyom
Tardieu, Francois
Tatanov, Ivan
Tataranni, G
Tate, Jennifer A.
Taugourdeau, Olivier
Tauriello, Gerardo
Tausz, Michael
P0426
SYM012
KNS01
P0350
P0353
P0439
P0651
SYM049A
SYM099B,
SYM104A,
SYM157, P0115,
P0416
P0687, P0688
SYM067
SYM034, P0176,
P0310
SYM011
SYM045
P0386
SYM006
SYM060A
SYM090
SYM156
SYM050
P0986
SYM086, SYM169
P0569
SYM172
P0702, P0704
SYM164
P0053
P0987, P0988
731

Author
Telford, Ian R. H.
Temponi, Livia
Temsch, Eva
Teng Seah, Koh
Teodoro, Grazielle
Teodoro, Grazielle
Terada, Kazuo
Terentieva, Elena
Terrab Benjelloun,
Anass
Terry, Janet
Tessler, Michael
Reference
SYM084, P0989
P0190, P0689
P0092
P0690
P0191
P0192
P0885
P0990, P0991
P0993
Tester, Mark
Testo, Weston
Thadeo, Marcela
Theim, T
Theodoulou, Freddie
Thibaud, Marie
Christine
Thiele, Kevin
P0121
P0994
SYM035, P0326
P0193
SYM106
SYM011
SYM070
SYM052
Thielen, Marc
Thien, Leonard
Thiers, Barbara
SYM137, P0209,
P0853
SYM151
SYM080
P0995, P0996,
P0997, P1002
KNS01 Thiet, Vu Minh
Thilakarathne,
Chamindathee Lakmini
Thiombiano, Adjima
Thiv, M.
SYM034, P0176,
P0194
P0782, P1037
P0928
Thode, Veronica
Thomas, Dane
P0974
SYM037
Thomas, Daniel Caspar SYM086, P1074
Thomas, Mark P0595
Thomas, Phil SYM119
Thomas, William Wayt SYM091, P0997,
P1002
Thompson, Helen P1075
Thompson, Ken SYM018
Thomsen, M. A.
Thornhill, Andrew
Thorp, Grant
Thórsson, Ægir
Thrall, Peter H
Thulin, Mats
Thumma, B. R.
Thurston, S.
Tian, Yu
Tibbits, J.
SYM011
SYM131
P0470
P0720
SYM055, SYM078,
SYM127
SYM089A
SYM037
SYM011
SYM030
P0393
732
Author
Tikhomirov, Natalia
Till, Walter
Tille, Stefanie
Reference
P0380
SYM004, P0730
SYM051
Tilney, Patricia
Tineo, J
Tingey, Scott
Tissue, David T.
Tokairin, H
Tokida, Takeshi
Toledo, M.M.
Tolstikov, Vladimir
Toma, Cezary
Tomio, Yamaguchi
Tomlinson, Vickey
Tonaco, Auria
Torabi, Masoud
Toriyama, Kinya
Torke, Benjamin M.
Tosh, James
Tosolini, A.-M.P.
Trad, Rafaela J.
Tran, Linh
Traoré, Salifou
Trapnell, Dorset
Trávníek, Pavel
Tremetsberger, K.
Trevaskis, Ben
Trivedi, Clare
Troleis, Juliana
SYM079A, P1076
SYM011
P0589
SYM016, SYM020,
SYM049B
P0272
SYM034
P0040
SYM069B
P1077
P1065
P0103
P0196
P0571, P0692
SYM032, P0635
SYM089B
SYM088B
P0887
P0718, P0998
SYM063
P1037
SYM171
SYM105A
SYM126
SYM032, SYM152
P0121
P0656
Trudgen, Malcolm P1069
Trujillo-Argueta, Sonia P0042, P0197
Truta, Elena
Tsai, Chi Chu
Tseng, Chryon-Man
Tshisikhawe, Milingoni
Peter
Tsou, Chih-Hua
Tsubota, Hiromi
Tsuchimatsu, Takashi
Tsukahara, Sayuri
Tsukaya, Hirokazu
Tsuneki, Shizuka
Tsurumaki, Y.
Tsutsumi, Chie
Tuckett, R.E.
P0585
P0985, P0999
P0693
P1080, P1096
P0693
P0861, P1000
SYM126
P0428
SYM066, SYM071,
P0629
P1001
P0559
P1078
SYM090

Author
Tudor, Richard L
Tugay, Osman
Tulig, Melissa
Reference
SYM070
P0785
P1002
Tuomisto, H.
Turnbull, Colin
Turnbull, Johanna D.
Turner, Barbara
Turner, Benjamin
Turner, Hubert
Turner, Shane
SYM116B
SYM069B
SYM022, P0091
SYM168, P0947,
P1042
P0910
SYM110
Turska, Anna
Tuskan, Gerald A
Tuteja, Narendra
Twyford, Alex D.
Tyberghein, Lennert
Tyerman, Stephen
SYM166, SYM167,
P0092, P0198
P0119
PL06
SYM032
SYM077, P1074
SYM075
SYM049A
Tyquin, Veronica P1003
U&#287;Urlu, Zübeyde P1084
U. Fangel, Jonatan
U.A. Ukrainskaya,
Ubierna Lopez, Nerea
Uchida, Kenichi
Uchida, Mayumi
Uchimiya, Hirofumi
Uchiyama, Hiroshi
Udovicic, Frank
SYM046
P0706
SYM050
P0572
P0522
SYM145, P0271,
P0329
SYM145
SYM018,
SYM088A, SYM100
SYM061, P0561
P0275
Ueda, Haruko
Ueda, Masafumi
Ueda, Minoru
Ueda, Takashi
Uehara, Ayumi
Uematsu, C.
Uemura, Shuhei
Ulloa, Carmen
SYM040
SYM042, P0628
P0607
P0411, P0426
P0560
P0199
P0572, P0558
P0179
Umehara, Mikihisa
Uniyal, P. L.
Uotila, Pertti
Urdampilleta, Juan D.
Uren, Victoria
Urn, Vladislav
Ursula, Ladinig
Ushijima, Kiyoharu
Ushijima, Tomiko
Usui, Yasuhiro
P0960
P0718
P1066
SYM105A
P0824
P1055
P1055
SYM034
Author
Utteridge, Timothy
Uysal, Tuna
Uyttewaal, Magalie
Uzunov, Dimitar
V. Neri, Andreza
V. Tinti, Bruno
Váa, Jií
Reference
SYM166
P0785
SYM065
SYM011, P0030,
P0752, P0753
P0232
P0232
SYM118
Vacha, Frantisek
Vaillancourt, Rene E.
P0675
SYM037, SYM121,
SYM149, P0423
P0787 Valcárcel, Virginia
Valdez-Leal, Juan P0057
Valiejo-Roman, Carmen SYM079A, P0990,
P0991
Valladares, Fernando SYM020, SYM164
Vallejo-Roman, Carmen SYM079
Van Aken, Olivier
Van Dam, Nicole
Van De Peer, Yves
Van Den Berg, Cássio
Van Den Berg, E.
Van Der Bank, Michelle P0774
Van Der Merwe, C.F. P1076
SYM092 Van Der Merwe,
Marlien
Van Der Niet,
Timotheus
Van Der Putten, Wim
Van Der Straeten,
Dominique
Van Dyk, Maria M
Van Ee, Benjamin W,
SYM063
SYM010, SYM036,
SYM096
PL06
SYM072, P0816,
P0913
P0191
SYM139
SYM036
SYM071
SYM037
SYM004, SYM081
Van Herwaarden, A F SYM160
Van Lijsebettens, Mieke SYM071
Van Rooyen, MW
Van Staden, Johannes
P1096
SYM145, P0930
Van Veen, H
Van Welzen, Peter C.
Van Wyk, A.E.
Van Wyk, B-E.
Van Wyk, Ben-Erik
Vandeleur, R K
Vanex, Fred
Vargas, M.A.
Vargas, Pablo
Vary, Laura B.
Vasco, Alejandra
Vashist, D
SYM012
SYM110, P0989
SYM117A, P1076
SYM079, SYM129
SYM079A,
SYM089, SYM129
SYM049A
KNS01
P0833
P0787, P1052
P0953
P0931
SYM012
733

Author
Vaughn, Matthew
Vaughton, Glenda
Ve, Thomas
Veerambakkam,
Ramachandran
Veeraputhiran, V
Reference
KNS01
SYM015
SYM055
P0200
Vekemans, Xavier
Veloso, Tomás
Veloz, Alberto
Veneklaas, Erik
Venter, Andor
Venter, S.
Vera-Batista, C
Verboom, George
Anthony
Verboven, Pieter
P0322
SYM126, P0429
P0020, P0148
P0889
SYM024, P0130,
P0743, P0965
SYM168
SYM108
P0610
SYM091, SYM170
Verbruggen, Heroen
Vergara-Silva,
Francisco
Versieux, Leonardo
Veruska Da Silva
Muniz, Ana
Vesk, Peter A
Vi, L.S.
P0569
SYM075
P0694
SYM004
SYM157
Viana, Pedro
Viccini, Lyderson
SYM018
SYM071
P1006
SYM157
Victoria-Hernandez, A. P1067
Vieira Santello, Gabriel P0138
Vieira, Eliane Do Rocio P1094
Vieira, R. C. P0715
Vieira-Neto, João
Vielle-Calzada, Jean-
Philippe
Vilalba-Ferreira, Carla
V.
Villanueva-Garcia,
Claudia
Villar, J. L.
P0364, P0367
KNS01
P0616
P0057
P0833
Villarreal, Juan Carlos
Villaseñor, J. L.
Villaverde, T
Vinall, K
Vinall, Kerry
Vincent, Olivier
Vincini, L F
Vincze, Eva
Visser, Vernon
Vivanco, K
Vladimirov, Vladimir
Vockenhuber, Elke
SYM087, SYM155
P0948, P0949
SYM085
SYM030, SYM031
SYM030
SYM151
SYM157
SYM032
P1066
SYM011
SYM094
SYM036
734
Author
Voelckel, Claudia
Voesenek, Laurentius
A.C.J.
Vogel, H.
Vogel, John
Vogl, C.
Voigt, Dagmar
Voinnet, Olivier
Von Aderkas, Patrick
Von Balthazar, Maria
Von Caemmerer,
Susanne
Von Konrat, Matt
Voothuluru, P
Vorontsova, Maria
Vovk, Myroslava
Voznesenskaya, Elena
Vrijdaghs, Alex
Vyas, Niral
Wabale, Hemant S.
Wafula, Eric
Wagatsuma, Takahiro
Wagner, Natascha
Wagner, Sarah
Wagner, W.L.
Wagstaff, B.
Wagstaff, Steven J.
Wahlberg, N.
Walder, Florian
Walker, Mandy
Wallace, Ian
Wallace, L.E.
Waller, Jade L
Wallis, Ian
Wallis, L
Wallnoefer, Bruno
Walls, Ramona L.
Walsh, Cherie
Walsh, Neville G.
Walts, Brandon
Waman, Mohan
Wanderley, Maria Das
Graças
Wang, Aiying
Wang, Bin
Reference
SYM097
SYM012
SYM039
P0540
P0811
SYM151
KNS01
SYM058
SYM099A, P0695
SYM028
SYM118, SYM155
SYM046
SYM100
P0697
SYM028, SYM068,
SYM127, SYM170,
P0225, P0255, P0696
SYM091
P1097
P0318
P0386
P0391
SYM004, SYM103,
P1011
P0804
SYM074
P0887
SYM104A,
SYM108, P1008,
P0859, P1008
SYM116B
SYM124
P0448
PL10
SYM074
SYM045, P0374
SYM037
SYM104B
SYM168, P0947
SYM048
SYM059
SYM084, P0892,
P0935
SYM038, P0377
SYM018, P0204
SYM002, SYM004
SYM049B
SYM155

Wang, Renqing
Wang, Rui-Jiang
Wang, S.-Z.
Wang, Shi-Jun
Wang, Xin
Wang, Xiu-Jie
Wang, Yan
Wang, Ye
Wang, Yu Hua
Wang, Zhong-Hua
Wanke, Stefan
Wanntorp, Livia
Ward, H.
Ward, Josephine
Wardle, Glenda
Author
Wang, Chuang
Wang, Chun-Neng
Wang, Co-Shine
Wang, Hsiang-Hua
Wang, Jiancheng
Wang, Lei
Wang, Li
Wang, Liangmin
Wang, Ming
Wang, Ming-Bo
Wang, Ning
Wang, Qiuyu
Wang, Qizhi
Wang, Quanxi
Wang, Ren
Ware, Doreen
Wargent, Jason
Warren, Ben
Warwick, Nigel
Washington, Jennifer
Marie
Wasley, Jane
Wasson, Anton
Watanabe, Chiaki
Watanabe, Masao
Waterhouse, Peter
Waterman, Melinda
Waters, Daniel
Waters, J.M.
Waters, Mark
Reference
P0557
SYM095, P0863
P0634, P0652
P0034
P0236
P0207
SYM095
P0205
P0596, P1050
SYM060B
SYM062
P1009
P1050
P0615
P0337
P0069
P0432
P0811
SYM027
SYM094B
SYM052, SYM060
SYM059
P0206, P0207
SYM153
SYM060
SYM080, P0804
SYM072
P0062
SYM083
SYM014, P0233,
P0234, P0936, P1062
SYM161
SYM023
SYM057
SYM089B
P0340
SYM022, P0091
SYM036
P0575
P0273
SYM060B
SYM023
SYM088
P0904
SYM153
Weisskopf, C
Welcker, Claude
Weller, James
Weller, S.G.
Wellman, Charles
Welsby, J
Welt, Rachel
Wen, Fang
Wen, Jun
Wendel, Jonathan F
Wendt, Tania
Wertman, Jaime
Wesenberg, Dirk
Wester, Petra
Westoby, Mark
Weston, Leslie
Weston, P.H.
Weston, Peter H.
Author
Waterway, Marcia
Watkins, James
Watkins, Jr., James
Watling, Jennifer
Watson, D.M.
Watson, David
Watson, Mark F
Watt, Michelle
Waugh, Robbie
Waycott, Michelle
Webb, Hamish
Webber, Bruce L.
Weber, A
Weber, Marjorie G.
Webster, Hollie
Webster, M
Weeks, Andrea
Wege, Juliet A
Weidenhamer, Jeffrey
Weigelt, Patrick
Weiler, Georg
Weising, Kurt
Weststrand, Stina
Weton, Peter
P0211
SYM008
SYM120, SYM172,
P0891, P1012
PL08
SYM002
SYM070
P0509
SYM058
SYM014, SYM092,
P0220
SYM051
SYM104, SYM171
SYM057, SYM099,
SYM104, SYM171,
P1062
P1013
SYM020
SYM037
P0208
SYM067
SYM150
SYM160
P0655
SYM115
P0209
Reference
SYM085
SYM116
P0193
SYM001, SYM156,
P0073
SYM001, P0532
SYM001
SYM005
SYM036, P0540,
P1098
SYM059
SYM116B
SYM051
SYM144A
P0592
SYM004, SYM103,
P0430, P1010 P1011
SYM151
P0439
P0643
SYM074
SYM086
SYM118
735

Author
Whalen, Molly
Wheeler, Janet
Whelan, James
Reference
SYM150, P0212
SYM153, P0341
SYM063, P0557,
P0620
SYM089B
SYM134
Whinder, Frances
Whitbread, Greg
White, Matt
White, Rosemary
Whitehead, Michael
Whitham, Thomas
Whitlock, Raj
Whitten, Mark
Whitten, W.M.
Wicke, Susann
Wickett, Norman J
SYM170
SYM060A,
SYM060B
P1014
SYM149
SYM091
SYM117
SYM142
Wiegand, Kerstin
Wieringa, JJ
Wieser, H.
SYM038B
SYM087,
SYM117B,
SYM155, P0386
SYM144A
P1045
SYM034
Wildova, Radka
Wilf, Peter
Wilkerson, Curtis
Wilkin, Paul
Wilkins, Katie A
Will, M.
Williams, J. H.
Williams, Joseph
Williams, Matthew
Williams, Nicholas
Williams, Norris
Williams, Paul
SYM025
SYM092, SYM101,
SYM121, P0048,
P0820
SYM059
P1015
SYM070
P0771
SYM090
SYM062
SYM046
SYM018
SYM117
P0035
Williams, Simon SYM055
Williams, Thomas C.R. SYM156
Willis, John
Willmann, Matthew R
Willyard, Ann
Wilson, Annette
Wilson, Gary
Wilson, Karen L.
P0961
SYM066
SYM074
P1075
P1016
SYM085, SYM091,
SYM168
SYM099, SYM131
SYM059
Wilson, Peter G.
Wilson, Sarah
Wilton, Aaron
Windham, Michael
Wink, Michael
Winkworth, R.C.
SYM118, SYM137
SYM088B, P0716
SYM129
P0747
736
Author
Winship, Lawrence
Winter, Klaus
Wirtz, Markus
Wist, T J
Wockner, Kimberley
Wöhrmann, Tina
Wojciechowicz, M K
Reference
SYM062
SYM004, SYM017
P0620
SYM058
SYM034
P0430
P0437
Wojciechowski, Martin SYM089A
Wolfe, Lorne SYM025
Woltering, Ernst J.
Won, Hyosig
Wong, Chui E
Wong, Connie
SYM010
P0214, P0215,
P0577, P1017
SYM060
SYM023
Wong, Gane K.-S
Wong, Sie Chuong
Wongniam, S.
Wongprasert,
Thawatchai
Wood, Andrew J.
SYM119
P0431
SYM171
P0327
Wood, Ken
Woodfield, Helen
Woodland, Dennis
Woodward, Ian
Wowk, Myroslawa
Wright, Harrison
Wright, Ian
Wright, Jeff
Wright, M
Wright, S Joseph
Wright, Shane
Wright, Stephen
Wu, Biao
Wu, Chia-Ti
Wu, Chung-Shien
Wu, Hen-Ming
Wu, Ming-Jou
Wu, Wei
Wu, Wenying
Wu, Xianjun
Wu, Yingying
Wu, Yuhuan
Wurdack, Kenneth J.
Wyse Jackson, Peter
Wyse, Sarah
Xia, Bing
Xia, Huijun
P0457
P0863
SYM028
P1018
SYM050
P0697
SYM070
P0220
SYM037
P0164
SYM114
P0235
SYM096
P0386
P0369
P0432
SYM062
P1019
SYM157
SYM027
SYM060A
SYM046
P1020, P1021
SYM081
SYM011
P0235
P0337
P0698

Author
Xia, Ke
Xiang, Jenny
Xiangyu, Liu
Xiaodong, Jia
Xie, Hongyan
Xie, Xiaonan
Xie, Xionan
Xing, Yaowu
Xiong, Guangyan
Xiu, Hu
Xu, Bo
Xu, Shuqing
Xu, Tonga
Xu, Xun
Xuan, Yong Hao
Reference
P0033
SYM103
P0674
P0350
SYM126
SYM153, P0572
P0579
SYM164
SYM059
P0306
P0984
SYM169
SYM061
SYM038B, SYM107
P0849
Xue, Bine
Xue, G P
Xue, Jiayu
Xun, Xu
P1022
SYM160
SYM155
SYM161
Yabuya, Tsutomu
Yadav, Neelesh
Yadav, S.R.
Yadav, Sangeeta
Yadav, Shrirang
P0600
P1024
SYM090
P0298
SYM090, P0032,
P1023
P0559 Yaeshima, M.
Yagihashi, T.
Yahara, T
Yamada, Kanako
Yamada, Masashi
P0172
P0911
P1026
P0453, P0454
Yamada, Yukiko P0699
Yamagishi-Costa, Julia P1099
Yamaguchi, Katsushi
Yamaguchi, M
Yamaguchi, Shinjiro
P0453, P0454
SYM046
KNS03, P0558,
P0572
P0642
P0856, P0940, P0968
Yamaguchi, Takashi
Yamaguchi, Tomio
Yamakawa, Chiyomi
Yamakawa, H.
Yamamoto, Kaoru
Yamamoto, Rie
Yamashita, Jun
Yamashita, Naoko
Yamato, Katsuyuki
Yamauchi, Takaki
Yan, Cheng
P1028
P0559
P1029
P0342
P1072
P0840
SYM040, SYM042
P0455
P0207
Yen, Hsio-Rong
Yhan, Antonio
Yie, Se-Won
Yigang, Wei
Yilmaz, Gulden
Yin, Jing
Yin, Lin-Ke
Yin, Xiaofeng
Yin, Yun Long
Yiyuan, Zhang
Yoder, John I
Yoko-O, Takehiko
Yokota, Etsuo
Yokota, Takao
Yoneyama, Kaori
Yoneyama, Koichi
Author
Yan, Guijun
Yan, Pengcheng
Yan, Xiaoling
Reference
SYM159
P0402
SYM165
Yan, Yueming
Yanada, K
Yanai, Junta
Yanai, Osnat
Yang, Bicheng
SYM159
P0679
P0275
SYM066
SYM038A,
SYM107, P0434
P1078 Yang, Chih Kai
Yang, Honglan
Yang, Jianchang
Yang, Kuoh-Cheng
Yang, Li
Yang, Nannan
Yang, Ya
Yang, Yang
Yang, Yongping
Yang, Yuen-Po
Yang, Zhenbiao
Yanisarapunt, S.
Yano, Kentaro
Yano, Okihito
Yano, Sachiko
Yansen, Yansen
Yasukuni, Yohei
P0457
SYM036
P0034
SYM066
P0380
SYM081
P0218
P0891
P0967, P0985
SYM061
SYM171
P0273
SYM085, P1030
P0642
P0578
P0885
Yasumoto, Akiko A. SYM126
Yatabe-Kakugawa,
Yoko
P1029
Yembaturova, Elena Yu. SYM079
P0693
P0249
P0395
SYM008
P0351, P0359
P0348
P0206, P0207
P0660, P0700, P0701
P0345
P0674
P0386
SYM046
SYM061
P0572, P0579
SYM153, P0579
SYM153, P0572,
P0579
737

Author
Yong, Jean W. H.
Yong, Kien Thai
Yoon, In Sun
Yoshida, Keiichiro
Yoshida, M.
Yoshihama, Isao
Yoshihara, Toshihiro
Yoshioka, Hirofumi
Young, Andrew
Young, Erica
Young, Iain M
Yousefzadeh Boroujeni,
Laleh
Yu, Chao Guang
Yu, Hyejin
Yu, Jing
Yu, Yu
Yuhong, Zheng
Yukawa, T.
Yukawa, Tomohisa
Zacarias, A
Zachgo, S.
Zachgo, Sabine
Zaheri, Ahmad
Zahraeifard, Sara
Zakeri, Hamideh
Zakharova, Ekaterina
Zakravsky, Petr
Zamaloa, Maria C
Zamfirache, Maria
Magdalena
Zan, Qin
Zanis, Michael
Zanne, Amy
Zare, Golshan
Zeisler, Viktoria V.
Zeng, Fan Suo
Zeng, Songjun
Zeng, Y.
Zenkteler, E
Zenkteler, Elzbieta
Zenkteler, M
Zerzour, Rabah
Zhan, Ya-Guang
Zhan, Yaguang Zhan
Reference
P0967
P1000
P0456
P1001
P0552
P0630
P0271
P0560
SYM015, SYM127,
SYM165, SYM166
SYM026
SYM051
P0488
P0207
SYM069
SYM056,
SYM099B,
SYM104A
P1033
P0587
P0435, P0436
SYM009
SYM039
P0437
P0437
P0437
SYM065
P0348
P0435, P0436
P0345
SYM117A
P0749, P0869
SYM123
P0350
SYM105B
SYM105B, P1078
P0263
SYM042
P0443
P0347
P0584
SYM177
P1032
P0113
SYM101, SYM121
P0585
738
Zhang, Yiping
Zhang, Yong-Jiang
Zhang, Yuanming
Zhang, Yujun
Zhang, Zhi-Qiang
Zhang, Zhonghua
Zhao, Guifang
Zhao, Junbin
Zhao, Ying-Tao
Zhao, Youyi
Zheng, Lu
Zhixiang, Zhang
Zhong, Jinshun
Zhongren, Guo
Zhou, Changyong
Zhou, Linlin
Author
Zhang, Baichen
Zhang, Baocai
Zhang, Bo
Zhang, Daoyuan
Zhang, Faqi
Zhang, Hao
Zhang, Hongyu
Zhang, Jianhua
Zhang, Jingjuan
Zhang, Libing
Zhang, Qisen
Zhang, X
Zhang, Xian-Chun
Zhang, Y.-F.
Zhang, Yeting
Zhou, Yihua
Zhou, Zhekun
Zhou, Zhuo
Zhu Et Al., Rui-Liang
Zhu, Chunwu
Zhu, J
Zhu, Rui-Liang
Zhu, Xinguang
Zhu, Yan
Zhuang, Jie-Yun
Zhunusbayeva, Dina
Ziba, Jamzad
Zidorn, Christian
Zieminska, Kasia
SYM082
P0588
P0457
P0205
SYM102, P1043
SYM107, P0402
P1034
SYM082
SYM052, SYM060
P0596
P0206
P0674
SYM076B
P0350
SYM060B
SYM086
Reference
SYM069B
SYM052, SYM059
P1043
P0236, P0457
P0762
SYM036
SYM060A
SYM036
SYM160
SYM008
P0589
SYM058
SYM095, P0874
P0811
P0386
SYM052, SYM059
SYM164, P0033
SYM172
SYM118
SYM034
SYM046
SYM118, P0818,
P1035
SYM028, SYM034
SYM123
P0419
SYM034
SYM009, P1058
SYM145
P0220

Author
Zimmer, Elizabeth
Zinniker, David
Zizka, Alexander
Zizka, Georg
Reference
SYM116A
SYM006
P1037
SYM002, SYM004,
SYM017,
SYM077B,
SYM103, P0782,
P1011, P1037,
P1038, P1039,
SYM042
SYM139
Zozomová-Lihová,
Judita
Zuccarello, Giuseppe
Zuccarello, Joe
Zucchi, M.I.
Zukauskiene, Judita
Zukowski, Wlademar
Zwart, Alec
Zwieniecki, Maciej
Zybartaite, Lina
Zych, Marcin
SYM097
SYM075
SYM075
P0998
P0226, P0449, P0919
P0021
SYM165, SYM166
SYM049
P0226, P0449
P0683
Zobell, Oliver
Zollikofer, Christoph
P.E
Zomlefer, Wendy SYM109
Subject
Acacia
Acacia, allied biota
SYM127
SYM078
Acacia, molecular phylogenetics
Acacia , pollination biology and breeding system
SYM078
SYM078
Acacia, rhizobial associations SYM078
Acacia , wood structure SYM089
Acanthaceae
Acer
Acer,
, evolution
leaf form
Acrospermum adaptation, environmental
Adhatoda
Adriana
Aesculus , evolution
African Plant Database
African Plants Initiative
Agavaceae, bimodal karyotype
Agave
Agave , biofuel agroecosystems, nutritional quality
Alcantarea , molecular phylogenetics algae
SYM163
SYM115
SYM056
SYM027
SYM164
SYM163
SYM129
SYM115
SYM118
SYM128
SYM117
SYM017
SYM045
SYM35
SYM004
SYM018,
SYM026
SYM075 algae, macroalgal floras in
Southern Hemisphere
Allee effect in reproduction
Allium
Aloe sect. Purpuscentes
Aloe, molecular systematics
SYM102
SYM031
SYM117
SYM117
Reference Subject Reference alpine plants alpine plants, New Zealand
SYM022,
SYM167
SYM083
Alstroemeriaceae, phylogeny SYM106 alternative oxidase of nonangiosperms
Alyssum , phylogeny
SYM156
SYM097
Amaryllidaceae alkaloids
Amborella genome
Anacardiaceae
Anaphalis, phylogeny, biogeography andromonoecy in Apiales
SYM145
SYM038
SYM115
SYM120
SYM079
Aneuraceae angiosperm female gametophyte angiosperms, basal, flowers
SYM027
SYM064
SYM099 angiosperms, basal, genomics SYM080 angiosperms, early SYM090
Angraecum pollination animal - plant associations,
Cenozoic
Annonaceae
SYM057
SYM104
Annonaceae, climbing
Anomochloa inflorescence ant - plant mutualism anther tapetum
SYM086
SYM104
SYM076
SYM129
SYM160
SYM079 Apiaceae, fruit and molecular characters
Apiales, andromonoecy
Apiales, bark and wood anatomy
Apiales, evolution and biogeography
Apiales, floral venation
SYM079
SYM079
SYM079
SYM169
739

Subject
Apocynaceae subfamilies
Asclepiadoideae and
Secamonoideae
Apocynaceae subfamily
Periplocoideae
Apocynaceae subtribe
Metastelmatinae
Apocynaceae, tribes apomixis apomixis, evolution of
Reference
SYM072
SYM168
SYM072
SYM072
SYM162
SYM039
SYM070 Aponogeton, programmed cell death aquaporins
Arabidopis , control of growth patterns
Arabidopsis
SYM049
SYM071
Arabidopsis , actin filaments and cables
Arabidopsis , actins
Arabidopsis , adapting to changing climate
Arabidopsis , adaptive trait evolution
Arabidopsis, cell wall expansion
Arabidopsis , chloroplast development
Arabidopsis , drought stress
KN01, KN03,
SYM041,
SYM042,
SYM046,
SYM126
SYM061
SYM061
SYM069
SYM171
SYM059
SYM068
Arabidopsis, epigenetics
Arabidopsis , fertilization and link to disease resistance
Arabidopsis , formin
Arabidopsis
Arabidopsis
, heteroblasty
, karyogamy
SYM054
SYM123
SYM064
SYM061
SYM066
SYM062
Arabidopsis , phosphate starvation
Arabidopsis , plastid retrograde signals
SYM052
SYM063
Arabidopsis , sepal patterning SYM067
SYM069 Arabidopsis , transposable elements
Araceae
Araceae, calcium oxalate crystals
Araceae, dated tree
Araliaceae, floral venation
SYM156
SYM114
SYM106
SYM169
Araucariaceae, diversification SYM119
Arctic cryptic speciation SYM096
Arecaceae
Arecaceae tribe
Trachycarpeae, radiation
Arecaceae, diversity across scales
Arecaceae, diversity in rainforests
Arecaceae, phylogenetics
SYM077
SYM144
SYM109
SYM109
SYM109
740
Subject
Arecaceae, plastid DNA aromatic plants
Asclepiadaceae
Asclepias, genome
Asclepias, phylogeny
Reference
SYM109
SYM163
SYM038
SYM038
SYM072
Ascomycetes SYM027
Asparagales, phylogenetics, biogeography
Aspleniaceae, systematics,
Australasia
Asteliaceae, phylogeny, dispersal
Asteraceae
SYM117
SYM116
SYM117
Asteraceae subtribe
Espeletiinae
SYM025,
SYM039,
SYM102,
SYM126,
SYM127,
SYM157,
SYM170
SYM168
Asteraceae, basal S American SYM108
Asteraceae, checklist
Asteraceae, evolution
Asteraceae, floral nectaries
SYM118
SYM120
SYM058
Asteraceae, island evolution SYM120
Asteraceae, secondary metabolites
Atlas of Living Australia
SYM145
PU04, SYM134
Australian flora, evolution PL09
Austrostipa classification
Balanophoraceae
Balsaminaceae, habit shifts bamboos
Banks, Sir Joseph bark anatomy, Apiales barley
SYM100
SYM009
SYM069
SYM025
SYM133
SYM079
SYM013,
SYM032,
SYM152
SYM103
SYM077
Bayes-DIVA method
Begonia, hybridization and introgression
Begonia, limestone caves
Begoniaceae
Bignoniaceae phylogeny and biogeography
Biodiversity Heritage Library
(BHL) biodiversity, New Guinea bioenergy crop biofuels, Agave biofuels, cell wall polymers biogeographic patterns,
Australia
SYM008
SYM077
SYM077
SYM136
SYM166
SYM017,
SYM161
SYM045
SYM059
SYM104

Subject Reference biogeography, ancestral area reconstruction biogeography, Australiasian flora
SYM103
SYM104 biogeography, Austral-
Pacific lineages
SYM108 biogeography, Bromeliaceae SYM004 biogeography, disjunct endemics biogeography, Malesia
SYM103
SYM110 biogeography, Malesia, glacial refugia biogeography, molecular dating
SYM110
SYM103 biogeography, reconstruction methods
SYM103 biogeography, S Hemisphere SYM168 biomechanics of plant development biomimetic materials and structures bio-ontologies
SYM067
SYM151
SYM048
Blandfordiaceae, phylogeny, dispersal
SYM117
Boraginaceae, Guiana Shield SYM171 botanic gardens, reducing footprint of botanic gardens, role in time of climate change
Botanical Capacity
Assessment Project botanical tools
SYM082
SYM082
SYM140
Brassavola , colonization
Brassicaceae
SYM170
SYM171
SYM024,
SYM041,
SYM126,
SYM167
SYM069 Brassicaceae, fruit shape diversity
Brassicaceae, genetic models for reduction
Brassicaceae, genome size
SYM169
SYM097
Brassicaceae, nectar secretion SYM058
Brassicaceae, polyploids SYM097
Brassicaceae, systematics and evolution
Brassicales Map Alignment
Project
SYM097
SYM096
Brassicales, evolution and genomics
SYM096
Brassicales, paleopolyploidy SYM157
Brassiceae, breeding systems SYM097 breeding systems
Bromeliaceae
Bromeliaceae, diversity, evolution, biogeography
Bromeliaceae, subfamily
Pitcairnioideae
Bromus
SYM126
SYM002,
SYM017,
SYM045
SYM103,
SYM004,
SYM077
SYM004
SYM025
Subject
Bryophyte Tree of Life bryophytes
Reference
SYM122
SYM013,
SYM022,
SYM023
SYM158 bryophytes, ecology of bryophytes, phylogeny and biogeography
Bursera, Simaruba clade
Burseraceae, diversification, radiation
Buxaceae
SYM098
SYM114
SYM114,
SYM115
SYM169
C
4
/C
3
grasses, fire responses SYM050
C
4
crops cadmium
Caladenia
Caladenia , pollination, herbivory calcium oxalate crystals,
Araceae
Calendula , hybridization, polyploidy
CAM
SYM050
SYM024
SYM167
SYM057,
SYM169
SYM114
SYM120
CAM, Euphorbia
Camellia
Campanulaceae
Campanulaceae subfam.
Nemacladoideae capacity-building capacity-building, online mentors
Capsella , population diversity carbon dioxide, elevated
Cardamine
Carex
Carex, anatomy and phylogeny
Carex, chromosome evolution
Caryophyllaceae
SYM012,
SYM017,
SYM045
SYM081
SYM153
SYM172
SYM172
SYM011,
SYM140
SYM140
SYM082
SYM034,
SYM035
SYM126
SYM085
SYM091 cassava castor bean cave speciation cavitation resistance
Celastrales, inflorescences cell death, petals cell death, programmed cell growth mechanisms cell shape generation cell wall digestibility
SYM085
SYM025,
SYM074
SYM35
SYM032
SYM008
SYM049
SYM076
SYM070
SYM070
SYM065
SYM065
SYM035,
SYM160
741

Subject cell wall proteins, biosynthesis cellular dynamics celluose synthesis
Cenozoic animal - plant associations
Centaurea
PL10
SYM104
Cephalanthera , achlorophyllous variants
Ceratodon
SYM025
SYM105
SYM023
Ceratodon, sex chromosomes SYM098 cesium SYM031
Chamaecrista , leaf anatomy SYM089 change, environmental character states, importance of context chemical defences
Chenopodiaceae
Chenopodium
Reference
SYM046,
SYM059
SYM061
SYM016
SYM076
SYM35
SYM031
SYM031
Children's books
Chiloglottis,
Chlorella pollination
PU03
SYM073
SYM026 chloroplast differentiation chloroplast dimorphism, chenopods chloroplast, thylakoid membranes chromatin
SYM068
SYM068
SYM068
Cistaceae
Cladia, biogeography
Cladoniaceae
SYM123
SYM164
SYM093
SYM093
Cleome , glandular trichomes SYM170 climate /environmental change
KN05, PU01,
SYM016,
SYM020,
SYM022,
SYM023,
SYM026,
SYM034,
SYM035,
SYM036,
SYM050,
SYM069,
SYM077,
SYM152,
SYM156,
SYM158,
SYM160,
SYM164,
SYM167
SYM082 climate change, role of botanic gardens climate shifts, Pleistocene S
African
Clusa
SYM098
SYM017
Coffea SYM167 collections, sampling density SYM170
742
Subject
Colutea , phylogeny
Commiphora , Malagasy
Reference
SYM130
SYM115
Compositae - see Asteraceae conifers, fossils SYM007 conifers, phylogeny SYM007 conservation conservation, ex situ
PL03, PU02,
SYM127,
SYM158,
SYM166,
SYM167
SYM082,
SYM166
SYM011 conservation, Global Strategy for Plant Conservation conservation, in Brazil SYM140 conservation, re-introduction SYM166 conservation, volunteers SYM166
Convolvulaceae
Cortinarius , phylogeny crop breeding crop genomics crop productivity
Corybas, Corysanthes clade phylogeny cotton polyploidy
Crassulaceae
SYM165
SYM172
SYM105
PL08
SYM045
PL02
SYM162 crops crops, apomixis crops, future
KN06, SYM023,
SYM030
SYM011,
SYM013
SYM039
SYM032,
SYM034,
SYM160
SYM129 Crotalaria , molecular phylogeny cryptogams
Cryptolepis , biogeography, phylogeny
Cucurbitaceae, dual phloem systems
Cucurbitaceae, evolution in
Australia
Cupressaceae, phylogeny cuticle, multifunctional interface
Cyananthus , evolution cyanobacteria cyanogenic glucosides
SYM027
SYM168
SYM069
SYM084
SYM007
SYM151
SYM172
SYM026
SYM035,
SYM053
SYM114 Cycadaceae, species, RAPD analysis cycads, phytochrome date
Cycas , species relationships
Cyperaceae tribe Cariceae
Cyperaceae tribe Schoeneae
Cyperaceae, biogeography and phylogeny
SYM119
SYM114
SYM085
SYM091
SYM091

Subject
Cyperaceae, mapaniids
Cyperaceae, online monograph
Cyperus , taxonomy
Cypripedium , pollination biology
Cytisus , phylogeny
Dactylanthus
Dactylorhiza , allopolypoly
Darwin, Charles
DECLIC defense strategies
Reference
SYM091
SYM091
SYM091
SYM142
SYM129
SYM009
SYM105
SYM142
SYM088
SYM070 dense sampling, Cyperaceae SYM091 descriptive data mark-up SYM136 desiccation tolerance development and RNAs
SYM002,
SYM054,
SYM116
SYM071 developmental genetics developmental modelling
Dicranum, phylogeography
SYM040
SYM067
SYM098
DigR, root system simulation SYM067
Dillenia SYM165
Dilleniaceae SYM165
Diospyros , New Caledonia SYM168
Dipterocarpaceae phenology SYM082
Disa pollination disease resistance dispersal syndromes
SYM057,
SYM073
SYM055,
SYM060
SYM164 dispersal, long distance dispersal, vicariance,
Myrtales
Diuridae, phylogeny and pollination diversity, divergence
DNA barcoding
SYM014
SYM108
SYM057
DNA replication
DNA, repetitive
Draba, speciation
Dracophyllum , diversification drought effects
KN10, SYM171
KN09, SYM088,
SYM114
SYM032
SYM038
SYM096
SYM108
Dryopteridaceae
Dryopteris , reticulate evolution
Ebenaceae, speciation ecological traits
SYM024,
SYM034,
SYM056,
SYM160
SYM008
SYM116
SYM168
SYM014
Subject economic plants eco-physiology ecosystem genetics education, botanical
Reference
SYM161,
SYM162
KN04
SYM128
SYM011 eFloras SYM137 embolism resistance and refilling
SYM049
Endeavour voyage specimens SYM133
Endeavour website endemism endoplasmic reticulum
SYM133
SYM002
SYM061
Epidendrum epigenetics
Eremophila
Ericaceae
, speciation
Ericaceae, Australasian phylogeny
SYM104
Ericaceae, mycoheterotrophic SYM124
Ericales, floral development SYM099
Erodium
SYM105
KN01, SYM123,
SYM162
SYM166
SYM108
Erysiphe eucalypts
SYM020
SYM024
SYM016
SYM049 eucalypts, canopy conductance eucalypts, carbon and water fluxes eucalypts, ecological tolerances eucalypts, genetic diversity, mating systems eucalypts, genomes eucalypts, hybridization eucalypts, secondary metabolites eucalypts, wood formation
Eucalyptus
Eucalyptus globulus
Eucalyptus grandis
SYM049
SYM121
SYM121
SYM037
SYM121
SYM037,
SYM121
SYM037
SYM167
SYM128
PL06, SYM037
SYM121 Eucalyptus subgenus
Eucalyptus
Eucalyptus, evolution
Eucomis pollination, scent compounds
Eugenia , phylogeny
Euphorbia global inventory
Euphorbiaceae
SYM121
SYM058
SYM131
SYM081
European e-Floras Initiative evo-devo, developmental process within species evo-devo, fruit shape in
Paysonia
SYM035,
SYM081,
SYM129
SYM137
SYM069
SYM069
743

Subject evolutionary history
Reference
SYM172 extinction extinction, mass, Pleistocene sclerophyll species
Fabaceae
SYM018
SYM104
SYM032,
SYM034,
SYM036,
SYM074,
SYM129,
SYM153,
SYM161,
SYM103
SYM089
Fabaceae, biogeography
Fabaceae, systematics and phylogeney
Fagaceae female gametophyte in earlydiverging lineages fern venation
SYM164
SYM080
SYM056 ferns SYM027,
SYM129 ferns, biology, biogeography SYM116 ferns, desiccation tolerance SYM116 ferns, DNA barcoding ferns, IndoPacific diversity ferns, IndoPacific filmy ferns, leaf form, function ferns, leptosporangiate, diversification ferns, lindsaeoid ferns, stomates
SYM088
SYM095
SYM095
SYM116
SYM086
SYM116
SYM116 fire flax rust resistance flax, transgenerational inheritance flooding stress
SYM014,
SYM077
SYM055
SYM123
SYM012
Flora of North America SYM137 floral develeopment, Ericales SYM099 floral evolution floral form in Zingiberales floral form, raindrop influence floral morphology floral traits, convergent
SYM099
SYM069
SYM102
Floras, next generation
Flore d'Afrique Centrale florigen flowering flowering plants, evolution food security
Forage sorghum forests, Neotropical dry
SYM099
SYM099
SYM137
SYM137
KN03
KN03
KN05, PL01
KN06, KN08
SYM024
SYM077
744
Subject form and function, evolution of
Forsterella , biogeography fossil, Protoananaceae fossils
Reference
SYM069
SYM103
SYM004
SYM006,
SYM007,
SYM027,
SYM086,
SYM094
SYM104
SYM101 fossils, Australasia fossils, Cenozoic paleofloras
S Hemisphere fossils, conifers fossils, Eucalyptus fossils, ferns fossils, flower origin fossils, Gnetales fossils, lycopods fossils, monocot
SYM007
SYM121
SYM092,
SYM116
SYM099
SYM119
SYM116
SYM106 fossils, Myrtaceae pollen fossils, Neotropical
SYM131
SYM077
Fragaria freeze/thaw-induced embolism
Fritillaria genome
SYM038
SYM049
SYM038 fungi SYM013,
SYM024 fungi, Australian macrofungi SYM093 fungi, biogeography S
Hemisphere fungi, ectomycorrhizal, saprotrophic
SYM093
SYM093,
SYM172 fungi, New Zealand endemics SYM093 fungi, sequestrate
Australasian fungi, symbioses
SYM093 fungi, tulasnelloid
Fusarium
SYM027,
SYM051,
SYM087
SYM027
KN08 gametophyte, competiton gametophyte, evolution
Gelsemiaceae alkaloids
SYM102
SYM064
SYM145 gene tree incongruence genera, problem of large genes, housekeeping
SYM074
SYM072
SYM071 genomes PL06, PL08 genomes, basal angiosperms SYM080 genomes, Brassicales SYM096 genomes, crop, wild relatives SYM038,
SYM161 genomes, digital library SYM038

grasses, cell wall polysaccharides grasses, DNA barcoding grasses, molecular basis of inflorescence evolution grasses, PPAM clade grasses, spinifex
Grevillea
GSPC Target 1, Plant List
Gymnadenia, multiple cytotypes gymnosperms, Tree of Life gypsophytes halophytes
Haplophyllum
Haptanthaceae
Haptanthus, new findings
Helianthus, hybridization, adaptation hepatics, morphological evolution herbarium specimens herbarium specimens online herbarium specimens, Flora of Nepal herbarium specimens, Hinton historic herbarium, Paris
Subject genomes, invasive populations genomes, plant evolution geometric morphometrics
Geonoma
Reference
SYM038
SYM048
SYM139
SYM077
SYM020 Geraniaceae
Geraniaceae genome SYM038
Gesneriaceae, Guiana Shield SYM171
Gesneriaceae, Neotropical
Global Compositae List
SYM077
SYM118
Global Plants Initiative SYM128
Glycine
Glycine, genome incongruence
Gnetales, phylogenomics, biogeography
SYM034,
SYM038
SYM074
SYM119
Goodeniaceae, diversification SYM108
Gorteria, pollinator diversity SYM102
Gossyium
Gramineae - see Poaceae
PL08
GrassBase grasses grasses, C
4
evolution
SYM100
SYM165
SYM050
SYM059
SYM088
SYM076
SYM155
SYM005
SYM005
SYM005
SYM005
SYM005
SYM100
SYM104
SYM015
SYM118
SYM105
SYM119
SYM167
SYM030
SYM171
SYM169
SYM169
SYM120
Subject herbarium, virtual herkogamy heterotrophic metabolism in plants heterotrophy in orchids
Hieracium , apomixis
Reference
SYM128
SYM015
SYM156
SYM105
SYM039
Hieracium
Hordeum
, reticulate genus
Himantoglossum homeostasis
evolution
Hookeriales, evolution
SYM170
SYM105
SYM156
SYM122
SYM013,
SYM032,
SYM152
SYM087 hornworts conservation hornworts, evolution and biology
SYM087 hornworts, stomatal evolution SYM087 houskeeping genes SYM071
Hoya phylogeny
Huperzia hybridization hybridization and species radiation hybridization, genomic footprint hybridization, New Zealand flora
Hydatellaceae hydraulic systems in plants
Hypoxidaceae, phylogeny, dispersal incongruence, gene trees, morphology
Indigofera, Cape Clade inflorescence development
SYM038,
SYM157
SYM094
SYM090
SYM049 hydraulic traits SYM014 hydrogen cyanide production SYM024
SYM049 hygroscopic particles on leaves
Hypnales, rapid evolution
Hypochaeris
SYM122
SYM126
SYM117
SYM170
SYM129
SYM076,
SYM152
SYM076 inflorescence diversity inflorescence ecology inflorescence evolution inflorescences, development of terminal flower insect - plant codiversification
Integrated Taxonomic
Information System (ITIS)
International Plant Names
Index (IPNI) intraspecific variation
SYM072
SYM116
SYM025,
SYM126
SYM083
SYM076
SYM076
SYM076
SYM078
SYM134
SYM134
SYM128
745

Subject invasion ecology
Reference
KN07, SYM018,
SYM025,
SYM165
SYM144 island biogeography, General
Dynamic Model island endemics SYM144 island endemics, Micronesia SYM144 island floras island plants, anagenesis
SYM144
SYM144
SYM144 island radiation, Arecaceae tribe Trachycarpeae islands, Mascarene colonizations islands, oceanic endemics
Isoetes
SYM144
SYM094
SYM012
SYM170 Jamesbrittenia
Japanese Knotweed
JSTOR Plant Science
SYM020
SYM128
SYM045 Kalanchoe
Kaliphora , fruit karrikin signalling keys, online interactive
Lanariaceae
SYM079
SYM153
SYM137
Lantana
Latin American Plants
Initiative
Lauraceae, model system leaf dissection
SYM117
SYM077
SYM128
SYM092
SYM066 leaf litter leaf margin characters leaf morphogenesis
SYM014
SYM164
SYM066 leaf shape development leaf unfolding leaf venation architecture legumes, African Genistoid
SYM066
SYM065
SYM056
SYM129 legumes, African Papilionoid SYM129 legumes, African Phaseoleae SYM129 legumes, African Psoraleoid SYM129
SYM089 legumes, Caesalpinoid classification legumes, Coluteoid clade legumes, Indigofereae
SYM130
SYM129 legumes, Iranian Papilionoid SYM130 legumes, Mimosoid classification legumes, Papilionoid phylogeny legumes, systematics and phylogeny legumes, tribe Hedysareae
SYM089
SYM089
SYM089
SYM130
746
Subject legumes, tribe Ingeae
Leguminosae - see Fabaceae
Reference
SYM089
Lejeunaceae, first hybrid
Lepidium , polyploidy
Lepidozia generic complex
SYM172
SYM097
SYM155 lichens lichens, biogeography S
Hemisphere
Liliaceae
SYM018
SYM093
Linum liverworts liverworts, biogeography, phylogeny liverworts, collections of
Stephani liverworts, genomes
Lomatia , speciation, phylogeography
Lotus genome lowland rainforests,
Neotropics
Lupin - wheat bread
Lupinus , biogeography
SYM038
SYM123
SYM158,
SYM172
SYM040,
SYM042,
SYM155
SYM005
SYM155 liverworts, spore wall ultrastructure
Lobeliaceae, diversification in Australia
SYM169
SYM084
Loganiaceae tribe Loganieae SYM108
SYM104
SYM161
SYM077
SYM153
SYM103
Lycium , dating, dispersal
Lycium , phylogeography, ploidy
Lycopersicon , evolution lycophytes
Magnolia
SYM107
SYM102
SYM069
SYM027
SYM009
Magnoliaceae maize
SYM009
SYM028,
SYM039,
SYM152
SYM161 maize genome
Malpighiales, floral structure, systemics
SYM099
Malpighiales, inflorescences SYM076
SYM099
SYM144
Malvaceae
Malvaceae subfamily
Dombeyoideae
Malvaceae tribe Malveae, gender dimorphism
Manihot maple leaf evolution mapping, predictive vegetation
Marchantia
Marchantia , developmental genetics
SYM104
SYM35
SYM056
SYM170
SYM040
SYM042

Subject
Marchantia,
Marchantia
Marsilea
Marsileaceae
Meconopsis
Medicago bioscreening
genome
genomics medicinal plants,
, transgenesis medicinal plants, India medicinal plants, Iran medicinal plants, Thailand
Medusagynaceae
SYM177
SYM177
SYM099 megaspore tetrads, competition
Melaleuca
SYM064
SYM025
Melaleuca , species complex SYM131
Melastomataceae
Merremia
Mesembryanthemum
SYM009
SYM165
SYM045
Reference
SYM040
SYM040
SYM166
SYM166
SYM126
SYM162
SYM009,
SYM145,
SYM163,
SYM177
SYM177 metabolites, bioactive
Michelia microbe - plant interaction
Mimulus
Minaria , evolution mistletoes modelling, species distribution molecules, ancient
Molluginaceae monocots, C
4
evolution
SYM153
SYM009
SYM016
SYM015
SYM072
SYM001
SYM144
SYM013
SYM017
SYM050
SYM059 monocots, cell wall biosynthesis monocots, ecology monocots, evolution, phylogeny monocots, fossils monocots, inflorescence types monocots, primary tissues monocots, transcriptomes morphometrics, ancestral floral shapes morphometrics, geometric mosses mosses, diplolepideousalternate mosses, haplolepideous mosses, phylogeny and biogeography
SYM106
SYM106
SYM106
SYM076
SYM069
SYM106
SYM139
SYM139
SYM022,
SYM023
SYM122
SYM122
SYM098,
SYM122
Subject
Muehlenbeckia, phylogeny
Murraya , diversification
Reference
SYM168
SYM171
Musa , phylogeography mycoheterotrophy names, retrieval names, starting points
SYM171
SYM124 mycoheterotrophy, parallel evolution of mycoheterotrophy, partial
SYM124
SYM124 mycorrhizal fungi SYM016,
SYM027,
SYM040 mycorrhizal fungi specificity SYM105,
SYM124
Myrtaceae SYM025
Myrtaceae, pollen
Myrtaceae, systematics, biodiversity
Myrtales, dispersal, vicariance name services
SYM131
SYM131
SYM108
SYM134
SYM136
Narcissus nectar evolution
SYM134
SYM015
SYM058 nectar function nectar, floral nectar, 'toxic' nectaries, extrafloral nectaries, floral, vascular supply
Nemacladus phylogeny
Neotropical diversity
Nicotiana, alkaloid metabolism
Nicotiana, allopolyploidy
SYM058
SYM058
SYM058
SYM129
SYM058
SYM172
SYM077
SYM053 nicotine biosynthesis nitrogen deficiency
Nothofagus nutrients, organic
Nyctanthes , sad tree
Nymphaea diversification in
India
Nymphaeaceae ocean acidification
SYM107
SYM053
SYM030
SYM158
SYM031
SYM003
SYM080
SYM080
SYM026
Ochnaceae, floral structure SYM099 online resources
(BiodiversityWorld and
KN07
Catalogue of Life)
Ophrys, floral isolation genes SYM169
Ophrys, osmophores
Opuntia
SYM142
SYM017
747

Subject
Orchidaceae
Orchidaceae, evolution, phylogeny
Orchidaceae, floral genetic diversity
Orchidaceae, mycoheterotrophic
Orchidaceae, pollination
Orchis, mycorrhizal associations
Orchis, pollination
SYM073,
SYM142
SYM105
SYM142
Orthophyllum SYM002
Oryza SYM012,
SYM028,
SYM032,
SYMO34,
SYM036,
SYM152,
SYM153,
SYM160
Oryza , cell wall biosynthesis SYM059
SYM060 Oryza , seeds, imprinted genes osmophores, Ophrys overgrowth mutants of barley and wheat
Oxalidaceae
Oxalis tuberosa
Reference
SYM045,
SYM167,
SYM169,
SYM171
SYM073,
SYM105,
SYM117
SYM117
SYM124
SYM142
SYM055
SYM011
SYM011
Pachycladon paleoecology palms, phylogenetics
Papaver
SYM097
SYM013
SYM109
KN04, SYM126
Papaver, self-incompatibility SYM070 papaya, sex chromosomes
Paphiopedilum, pollination biology
Paraphyly and systematics parasitic plants parasitic plants, genome parasitic plants, strigolactones
Paris Herbarium renovation
Parmeliaceae, origin
Passiflora
SYM096
SYM142
SYM094
SYM001,
SYM009,
SYM128
SYM038
SYM153
SYM005
SYM093
SYM129
Passifloraceae, Guiana Shield SYM171
Paysonia , fruit shape diversification
Pennisetum, apomixis
SYM069
SYM039
Persicaria SYM020
748
Subject pests and diseases
Petunia, self-incompatibility
Phaseolus
Reference
KN07, KN08,
SYM010,
SYM024,
SYM035
SYM062
SYM35 phenomics
Phenomics Ontology Driven
Data repository phenotypic plasticity
SYM048
SYM048
SYM020
Philodendron, thermogenesis SYM156 phosphate /phosphite signalling and homeostasis phosphate acquisition, deficiency photosynthesis
SYM052
SYM052
SYM012,
SYM017,
SYM028,
SYM034,
SYM050,
SYM162
SYM068 photosynthesis, chloroplast dimorphism in Kranz chenopods photosynthesis, Euphorbia SYM081
Phrymaceae SYM015
Physcomitrella, proteins
Physcomitrella, strigolactones phytochrome photosensory signalling
PhytoKeys ejournal phytoplankton
Pinaceae
Piper
Pisum
SYM048
SYM153
PL07
SYM136
SYM026
SYM007,
SYM025
SYM163
SYM032,
SYM036
SYM002
SYM075
Pitcairnia
Pithophoraceae
Plagianthus,
Plant List
dioecy SYM099
KN09, SYM011,
SYM118
SYM118 Plant List, hornworts, liverworts
Plant Ontology plants, art, Mexico
SYM048
SYM003 plants, disease and resistance SYM010 plants, early land plant genomes plants, fertilization
SYM155
PL05 plants, growth model plants, medicinal plants, modelling plants, mythology, art plants, relocation plants, trafficking pathways
SYM167
SYM177
PL04
SYM003
KN07
SYM042

Subject
Pleomele
Poa phylogeny
Poaceae
SYM100
SYM076
SYM100
SYM106
SYM100
SYM007
SYM064
SYM022
SYM062
KN04
SYM062
SYM015
SYM102
SYM072
SYM062,
SYM065
SYM131
Reference
SYM009
SYM100
SYM024,
SYM025,
SYM030-032,
SYM034,
SYM036,
SYM100,
SYM152,
SYM153,
SYM161,
SYM165
SYM100
SYM084
SYM057
SYM099
SYM057
SYM142
SYM057,
SYM142
SYM139
SYM123
SYM073
SYM165
SYM166
SYM015,
SYM057
SYM058
SYM102
SYM012,
SYM020,
SYM102,
SYM168
Poaceae subfamily
Chloridoideae
Poaceae subfamily
Danthonioideae
Poaceae, Agrostidinae
Poaceae, inflorescences
Poaceae, Koeleriinae
Poaceae, phylogeny
Poaceae, PPAM clade
Podocarpaceae evolution
Podostemaceae, megagametophyte evolution polar plants pollen - pistil interaction pollen allergens pollen biology pollen dispersal pollen structure pollen transfer, milkweeds pollen tube growth pollen, Myrtaceae, extant, fossil pollination by deception pollination, animal pollination, effect of weather on orchids pollination, Euglossine bees pollination, orchids pollinator shift hypothesis pollinator syndromes, epigenetic regulation of
Pollinators of orchids pollinators, bats pollinators, birds pollinators, insects pollinators, rodents pollinators, Roscoea
Polygonaceae
Subject
Polygonum
Polygonum, Indian
Himalayas polyploidy polyploidy and species radiation polyploidy in New Zealand flora polyploidy, orchids
Polypodiales
Polystichum in karst caves population dynamics
Populus
Populus transcriptome response
Portulacacinceae potato, genome
SYM164
SYM049
SYM017
SYM107
Primula , breeding system evolution programmed cell death
Prometheus Wiki
Proteaceae
SYM102
SYM010
SYM014
SYM015,
SYM020
SYM099 Proteaceae, floral traits
Proteaceae, phylogeny, diversification protein function, structural basis of protein profiles in nectar
SYM104
SYM055 protein, microtubule endtracking proteomes
Quercus
Quiinaceae
SYM058
SYM061
SYM032,
SYM068,
SYM156
SYM068
Psidium , phylogeny
Psychotria , anti-herbivore defense
Puya
SYM131
SYM114
SYM017
SYM164
SYM099
SYM092 rainforest floras, Australian, functional traits rainforest floras, Australian, origins rainforests, biomechanical diversity of
Ranunculus , apomixis
Ranunculus , hybridization
SYM092
SYM151
SYM039
SYM039 rare /endangered /threatened species recombination, role in incongruence
KN10, SYM127,
SYM167
SYM074 reintroduction of rare species SYM009
Reference
SYM020
SYM094
SYM127,
SYM157
SYM083
SYM083
SYM105
SYM027
SYM116,
SYM027
SYM008
SYM014,
SYM015,SYM127
749

Subject reproduction, outcrossing, selfing reproductive biology reproductive development reproductive diversity, ecology, evolution respiration resurrection plants retrograde signalling
Rhaphanus
Rhinanthus
Rhizobacteria
Rhizobia and Acacia rhizosphere interactions rhizosphere, N competition SYM051 ribosome in leaf development SYM071 rice breeding rice, nutrient content ricinoleic acid
Ricinus
SYM078
SYM051
SYM012,
SYM028,
SYM032,
SYM034,
SYM036,
SYM152,
SYM160
SYM028,
SYM032
SYM153
SYM032
SYM032
RNA signalling, intracellular and intercellular
RNA silencing
RNAs in plant development rock outcrops root architecture root exudates, bioactive root growth
Reference
SYM102
KN04, SYM015
SYM152,
SYM160
SYM102,
SYM169
SYM156
SYM054
SYM063
SYM126
SYM128
SYM036
SYM060
SYM060
SYM071
SYM002
SYM036
SYM051
SYM046,
SYM153
SYM153
SYM067 root system, simulation,
DigR root thermotolerance root: soil interactions
Rosaceae
Roscoea , reproductive biology
Rubiaceae
Rubiaceae tribe Rubieae
Rumex
Rumex , dioecy
Rutaceae
Rutaceae subfam.
Spathelioideae
SYM036
SYM036
SYM038
SYM102
SYM167
SYM094
SYM012
SYM102
SYM171
SYM115
750
Subject
Rutaceae subtribe Galipeinae floral whorls
Rutidosis
Saccharum
Salicaceae salinity stress salinity, progammed cell death
Salix
Sapindaceae
Sapindaceae, biogeography
Reference
SYM069
SYM127
SYM030,
SYM031
SYM164,
SYM165
SYM032
SYM070
SYM165
Sapindaceae, New Caledonia SYM115
Sapindales, systematics, evolution savanna
SYM115,
SYM171
SYM014 savanna biome, fire resistance
Schiedea, species tree
Schoenoxiphium
SYM056
SYM103
SYM077
SYM074
SYM085
Scrophulariaceae tribe
Limoselleae
SYM170 secondary growth, evo-devo SYM069 secondary metabolism seed ecology seed plant phylogenomics, phylogeny self-incompatibility
SYM053,
SYM145
SYM167
SYM119
SYM062,
SYM126,
SYM070
SYM151 self-irrigation by fog collection
Semantic Web
Senecio
Setaria
Sexual selection
SYM048
SYM126,
SYM157
SYM162
SYM015 shoot apex, ribosome role shoot apical meristem development shoot apical meristem, mechanical signals shoot maturation signalling molecules
Silene
SYM071
SYM067
SYM065
SYM066
KN03, SYM010,
SYM024
SYM025 soil biota soils, nutrient-poor
Solanaceae
Solanaceae, biodiversity, genomics
Solanaceae, fruit development
Solanaceae, phylogeny and biogeography
Solander, Daniel
SYM016,
SYM036
SYM041
KN02, SYM102
SYM107
SYM107
SYM077
SYM133

Subject
Solanum
Reference
KN02
SYM107 Solanum subgenus
Leptostemonum
Solidago
Sorghum soybean soybean gene phylogeny soybean genome soybean, shoot apical meristem
Spartina genome speciation species distribution modelling species lists species radiation, New
Zealand species tree estimation
SYM128
SYM035,
SYM162
SYM162
SYM034
SYM039
SYM038
SYM060
SYM157
KN02
SYM144
SYM134
SYM083
Sphagnum phylogeny
Spiranthes , allopolyploidy
Sporobolus , resurrection grass stem gravitropism, general model
Stipagrostis, fog-collecting
SYM074
SYM098
SYM105
SYM054
SYM067 stomatal density stomate evolution storage proteins strawberry genome strawberry, origin stress tolerance
SYM151
SYM164
SYM087,
SYM116
SYM032
SYM038
SYM038 stress, below-ground stress, effect on flower size stress, environmental stress, water
SYM024,
SYM041,
SYM160
SYM020,
SYM036
SYM164
SYM123 strigolactones sugarcane
SYM036
KN03, SYM153
SYM030,
SYM031
SYM120 sunflowers, hybridization, adaptation supertrees, palms
Swartzia , classification
SYM109
SYM089
Switchgrass systematics /taxonomy in
21st century
Syzygium
SYM161
KN09 tassel-ferns
SYM131
SYM116
Taxonomic Literature (Tl-2) SYM136
Subject
TaxonX markup language
Taxpub markup language
Reference
SYM136
SYM136 tea teaching about plants
Tecophilaeaceae
Telopea , speciation, phylogeography temperature stress
SYM153
SYM140
SYM117
SYM104
Theaceae
Thellungia , genome thermoregulation in plants
Thlaspi threatened species
Thysananthus , phylogeny
Tigridium magnifica
Tillandsia , evolution and phylogeography tobacco tobacco genome tobacco, allopolyploidy tomato, evolution tomato, leaf development
Trifolium genomics
Trithuria , breeding systems and biology
Triticum tropical tree diversity
Tropicos
SYM053
SYM157
SYM107,
SYM157
SYM069
SYM066 trace gas detection
Tragopogon genome evolution traits, functional, Proteaceae flowers transgene silencing transpiration transposable elements
Tree of Life, bryophytes
SYM010
SYM157
SYM099
SYM060
SYM049
SYM069
SYM122
Tree of Life, dating land plants tree structure traits
SYM086
SYM114 trichomes, glandular, Cleome SYM170
Trifolium SYM35
SYM162
SYM090
SYM013,
SYM030,
SYM032,
SYM034,
SYM036,
SYM152,
SYM160
SYM114
SYM134
SYM152
SYM153
SYM041
SYM156
SYM024
KN10
SYM172
SYM009
SYM004
Umbelliferae - see Apiaceae urban floras SYM018
751

Subject
Utricularia suction traps, functional morphology
UV radiation
Uvaria evolution
Vaccinium waratah water stress
Reference
SYM151
SYM023
SYM086
SYM022
SYM096 VEGI
Vella
Verbenaceae phylogeny and biogeography
SYM077 vernalization in Arabidopsis SYM060 vicariance, dispersal,
Myrtales
Viola, evolution, Hawaii
SYM167
SYM108
SYM172
Violaceae, Hawaii
Violaceae, pollen morphology
Virtual Endeavour website
Virtual Library of Biology
(vifabio)
Vitis vinifera
SYM172
SYM172
SYM133
SYM136
PU01
SYM020
SYM160
SYM070 waterlogging, programmed cell death wetlands SYM012
Subject wheat wheat genome wheat pathogens wine-growing areas
Wittrockiella wood anatomy, Apiales wood anatomy, pit structure wood formation, evo-devo woodland, Banksia
World Flora
Wurmbea
Xingxueanthus
X-ray tomography xylem embolism repair xylem refilling
Zea
Zingiberaceae
Zingiberales, floral form
SYM011
SYM015
SYM094
SYM099
SYM049
SYM049
SYM028,
SYM039,
SYM152,
SYM161
SYM102
SYM069
Reference
SYM013,
SYM030,
SYM032,
SYM034,
SYM036,
SYM152,
SYM160
SYM048,
SYM161
KN08
PU01
SYM075
SYM079
SYM049
SYM069
SYM015
752