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THE USDA FOREST SERVICE-RMRS FOREST FUNGI
COLLECTION: RESOURCE FOR FUNGAL IDENTIFICATION,
DEVELOPING BIOLOGICAL CONTROLS, PREDICTING INVASIVE
PATHOGENS, AND PREDICTING POTENTIAL IMPACTS OF
CLIMATE CHANGE
Sara M. Ashiglar, John W. Hanna 1, Amy L. Ross-Davis1, and Ned B.
Klopfenstein 1
ABSTRACT
The Moscow Forestry Sciences Laboratory
of the Rocky Mountain Research Station
(RMRS)
has a unique collection of
forest/tree- associated fungi with over
15,000 living specimens. Based in Moscow,
ID, this USDA APHIS-PPQ (Plant Protection
and
Quarantine)
containment
facility
houses fungal archives from approximately
35 states and 30 countries. The collection
involves the work of at least 50 USDA
Forest Service research scientists and
collaborators over the past 30 years.
Though the collection primarily focuses on
species associated with Armillaria root
disease from western North America, it
also contains fungi such as Phellinus,
Fusarium, Raffaelea, tree-root endophytes,
and other fungi associated with wood
decay. The RMRS Forest Pathology group
and collaborating scientists continue to use
and add to the collection for research on
prediction of invasive pathogens, climate
change, evolutionary relationships, and
identification
of
forest
DNA-based
pathogens.
Unfortunately,
long-term
maintenance of this invaluable collection is
not well funded and thus this collection is
In: Chadwick, K. Comp. Proceedings of the
61st Annual Western International Forest
Disease Work Conference; 2013 October 611; Waterton Lakes National Park, Alberta,
Canada. 1 USDA Forest Service, Rocky
Mountain Research Station, Forestry
Sciences Laboratory, Moscow, ID.
127
not
optimally
maintained.
With
an
estimated 25 percent of specimens with
lost viability in the collection, the RMRS
Forest Pathology laboratory continues to
seek
long-term
solutions
for
stably
maintaining the fungal archive collection.
BACKGROUND
Culture collections have changed history in
examples ranging from the development of
Penicillium-derived antibiotics to enzymes
from the thermophilic bacterium Thermus
aquaticus used in polymerase chain
reaction for molecular genetics (Dugan et
a/. 2011). In plant pathology, such
depositories
of
living
material
are
invaluable resources that aid the study and
control
of
plant
pathogens.
Often,
specimens held in culture collections allow
scientists to quickly identify, trace the
origin, control the spread and generally
assist in understanding the biology of
diverse existing and novel plant pathogens
(Kang eta/. 2006). Forest fungi play critical
environmental roles in almost every aspect
of forest ecosystems,
which
include
mycorrhizal
associates,
beneficial
decomposers, native and invasive plant
pathogens, and other biological agents.
These fungal activities have primary
influences on forest health and interact
with fire, water, climate change, and other
environmental
components
of
forest
ecosystems. Thus, a fungal collection that
is associated with geographic origin, host
trees, and various time points can
contribute to diverse aspects of forest and
grassland research.
The USDA Forest Service- RMRS, Moscow
Forestry Sciences
Laboratory
(MFSL)
houses one such culture collection. Its
fungal archive collection houses over
15,000 living specimens of forest fungi
including root-disease pathogens, fungal
endophytes,
fungal
saprophytes,
and
wood-decay
fungi
(Figure
2).
Our
laboratory is an APHIS Plant Protection and
Quarantine containment facility permitted
to receive and house forest fungi from
anywhere in the world. In addition, the
MFSL follows international conventions that
do not allow for the commercialization of
biological resources from international
sources.
Most cultures are from the genus Armillaria
(Figure 1) collected from the interior
western North America (e.g., Washington,
Oregon, Idaho, and British Columbia). In
addition to the North American Armillaria,
we have an extensive worldwide collection
of Armillaria isolates, as well as an
assortment of local and foreign wood-rot,
endophytic, and pathogenic fungi. Genera
include, though are not limited to Phellinus/
Fusarium, Heterobasidion and Raffae/ea.
The culture collection represents the work
of approximately 50 current and former
Forest
Service
employees
and
collaborators, houses isolates from more
than 30 countries and 35 states, and has
living collections dating back to 30 years.
Collections continue to be archived as the
laboratory conducts new research.
for research on 1) developing of DNAbased methods for identification of forest
fungi and other microbes (e.g., Nelson et
a/. in press;
Elias-Roman et a/. 2013,
Ross-Davis et a/. 2012, Stewart et a/.
2006 ), 2) determining evolutionary/genetic
relationships among fungi (e.g., Ross-Davis
et a/. 2012), 3) determining effects of
climate change on the distribution of forest
pathogens and associated fungi (e.g.,
Klopfenstein eta/. 2009, Klopfenstein eta/.
2011, Hanna et a/. 2012), 4) developing
methods to predict potentially invasive
pathogens
before
they arrive
(e.g.,
Klopfenstein et a/. 2011, Hanna et a/.
2012, Ross-Davis et a/., this proceedings
volume) 5) monitoring changes in forest
fungi over time, 6) identifying potential
biological control agents, and 7) numerous
other research topics (Figure 3).
CURRENT AND FUTURE USES
The Moscow Forestry Sciences Laboratory
(MFSL)
Culture
Collection
has
and
continues to support diverse research
disciplines related to the identification and
ecological roles of forest fungi. For
example, the Rocky Mountain Research
Station - Forest Pathology group and
collaborating scientists use the collection
Figure 1. Armillaria mushrooms in a forest
stand
declining
from
Armillaria
root
disease. Armillaria isolates from around the
world make up the majority of the MFSL
Culture Collection. Photo credit: John
Hanna.
128
7
Figure 2. Incubators and examples of fungal cultures on agar housed in the Moscow
Forestry Sciences Laboratory Culture Collection.
Figure 3. Example of how the MFSL Culture Collection is used. Specimens are generally
collected from the forest as fruit bodies, rhizomorhps, rotten wood, etc. and appropriate
accompanying data is collected on site (e.g., latitude, longitude, host, etc.). Fungi are
isolated onto agar-based culture medium, and molecularly identified using PCR and DNA
sequencing. This information may then be related back to the collection site through
modelling, analysis of gene expression, continued monitoring, comparison with other
specimens, and related research on forest fungi.
CURRENT STATUS AND GOALS FOR
THE CULTURE COLLECTION
Fiscal support of biological specimen
collections can be challenging in a scientific
world dominated by competitive grants
(Dugan et a/. 2011), and maintenance of
the MFSL fungal culture collection is no
exception. Unfortunately, no long-term
funding exists to maintain or update this
129
collection. An estimated 25 percent of
fungal archives may have already lost
viability. The database is in great need of
updating and the cultures themselves
require more stable archiving system (e.g.,
frozen cultures).
The forest pathology group at the MFSL
has been working with the United States
Culture
Collection
Network
(USCCN,
www.usccn.org) to update culture archive
techniques, create a backup set of cultures,
modernize database management, and
register the culture collection resource for
public acquisition. Still, while we are
assessing the tools to complete these
goals, progress remains limited by funding
support. We continue pursuing support for
long-term funding to hire a full-time
archivist/collection curator and provide
associated
culturing
supplies.
In
conjunction with the goals of the USCCN,
we hope to continue meeting national and
international requests for fungal isolates
and create a publicly available, online
resource
to
allow
easy
access
of
information pertaining to the culture
collection. Isolate requests continue to
increase over time, and indicate the
importance and recognition of the value of
this fungal collection. Finally, we continue
to update our databases for DNA-based
identifications of the fungal specimens,
including links to sequence submissions to
the international GenBank database . This
information
will
contribute
to
our
understanding of the diverse ecological
roles of fungi in forest ecosystems.
REFERENCES
Dugan, F.M .. A. Wiest, and K. McCluskey. 2011. Public germplasm collections and
revolutions in biotechnology. Journal of Bioscience 36(2):205-209.
Elfas-Roman, R.D. and others. 2013. Incidence and phylogenetic analyses of Armillaria
spp. associated with root disease in peach orchards in the State of Mexico, Mexico.
Forest Pathology 43(5):390-401.
Hanna, J.W. and others. 2012. A bioclimatic approach to predict global regions with
suitable climate space for Puccina psidii. Pp. 131-136 inS. Zeglen and P. Palacios
(camps) Proceedings of 59th Annual Western International Forest Disease Work
Conference, 2011 October 11-14, Leavenworth, WA. USDA Forest Service, Forest
Health Protection, Region 5. Portland, OR.
Kang, S. and others. 2006. Plant pathogen culture collections: it takes a village to
preserve these resources vital to the advancement of agricultural security and
plant pathology. Phytopathology 96:920-925.
Klopfenstein, N.B. and others. 2009. Approaches to predicting potential impacts of
climate change on forest disease: an example with Armillaria root disease. Res.
Pap. RMRS-RP-76. USDA Rocky Mountain Research Station. Fort Collins, CO.
Klopfenstein, N.B. and others. 2011. Approaches to predicting current and future
distributions of Puccinia psidii in South America under climate change scenarios.
Pp. 450-454 in J. on<;alves et al. (eds.) Proceedings: IUFRO Conference on
Improvement and Culture of Eucalypts; IUFRO Working Group 2.08.03, Porto
Segura, Bahia, 14-18 November, 2011. Piracicaba, Brazil.
Nelson, E.V. and others. 2013. First report of the Armillaria Root disease pathogen,
Armillaria gallica, on Douglas-fir (Pseudotsuga menziesii) in Arizona. Plant Disease
97:1658.
Ross-Davis, A.L. and others. 2012. Advances toward DNA-based identification and
phylogeny of North American Armillaria species using elongation factor-1 alpha
gene. Mycoscience 53:161-165.
Stewart, J.E. and others. 2006. Molecular characterization of Fusarium oxysporum and
Fusarium commune isolates from a conifer nursery. Phytopathology 96(10): 11241133.
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