Fundamental Aspects of Plant
Cryopreservation
17-18th February 2009
Royal Botanic Gardens, Kew,
Wakehurst Place, West Sussex, UK
Organisers: Jayanthi Nadarajan & Hugh W. Pritchard
PROGRAMME:
1
CONFERENCE PROGRAMME,
ABSTRACTS & PARTICIPANTS
CONTENTS
Page
Introduction to the sponsors
3
Programme
5
Abstracts:
Oral presentations
7
Poster presentations
List of participants
26
32
2
INTRODUCTION TO THE SPONSORS
European Science Foundation
The European Science Foundation (ESF) is an association of 80 member organisations devoted to scientific
research in 30 European countries. Since we were established in 1974, we have coordinated a wide range of panEuropean scientific initiatives, and our flexible organisation structure means we can respond quickly to new
developments. ESF's core purpose is to promote high quality science at a European level. The ESF is committed
to facilitating cooperation and collaboration in European science on behalf of its principal stakeholders (Member
Organisations and Europe's scientific community). This cross-border activity combines both 'top-down' and
'bottom-up' approaches in the long-term development of science.The Foundation is committed to providing
scientific leadership through its networking expertise and by ensuring that there is a European added value to all
of its initiatives and projects. [http://www.esf.org]
COST
COST is an intergovernmental framework for European Cooperation in Science and Technology, allowing the
coordination of nationally-funded research on a European level. COST contributes to reducing the fragmentation
in European research investments and opening the European Research Area to cooperation worldwide. The goal
of COST is to ensure that Europe holds a strong position in the field of scientific and technical research for
peaceful purposes, by increasing European cooperation and interaction in this field. This research initiative
makes it possible for the various national facilities, institutes, universities and private industry to work jointly on
a wide range of Research and Development (R&D) activities. COST – together with EUREKA and the EU
framework programmes – is one of the three pillars of joint European research initiatives. These three
complementary structures have differing areas of research. COST has clearly shown its strength in noncompetitive research, pre-normative cooperation, and solving environmental, cross-border and public utility
problems. It has been successfully used to maximise European synergy and added value in research cooperation
and is a useful tool to further European integration. Ease of access for institutions from non-member countries
also makes COST a very interesting and successful tool for tackling topics of a truly global nature.
[http://www.cost.esf.org]
COST ACTION 871- Cryopreservation of Crop Species in Europe
Plant germplasm stored in liquid nitrogen (-196°C) does not undergo cellular divisions. In addition, metabolic
and most physical processes are stopped at this temperature. Therefore, plant germplasm preserved under
cryogenic storage can be maintained for very long periods of time and problems that are typical for storage in
the active growth state, like genetic instability and the loss of accessions due to contamination, loss of vigour and
totipotency and human error during continual subculturing are overcome. So far, cryopreservation procedures
have been developed for the in vitro tissues and non-orthodox seeds of about 200 plant species. There are,
however, still a very limited number of examples in Europe where cryopreservation is used routinely for plant
germplasm conservation. This is mainly due to:
 the unavailability of efficient and robust cryopreservation protocols applicable to many plant species
and diverse germplasm types;
 limited awareness of plant researchers unacquainted to recent developments in cryogenic storage
methods;
 lack of coordinated research on plant cryopreservation.
To address these shortfalls, the goal of this COST Action is to create a network that brings together European
scientists with an expertise and/or interest in plant cryopreservation with the main aim of developing efficient
cryopreservation procedures.
Royal Botanic Gardens Kew
RBG Kew currently employs over 680 staff, over 60% of whom are specialist plant or fungal scientists or
botanical horticulturists. The mission of the Royal Botanic Gardens, Kew is: To inspire and deliver sciencebased plant conservation worldwide, enhancing the quality of life. Our business aim is to produce basic and
applied information about plant- and fungal-related topics and to manage and communicate this to all our
stakeholders. This aim is carried out through science and research in systematics, biological interactions,
economic botany, conservation and horticulture. This is underpinned by our extensive collections of living and
preserved plants and fungi, associated artefacts, literature and archives, and is interpreted to the public through
the Gardens at Kew and Wakehurst Place. These provide an amenity for the public, offering the opportunity to
learn about plants and our wider work. Our ability to increase our outreach and global impact depends on the
effectiveness of our communications. Our education programmes, dissemination activities, our national and
3
international partnership networks are fundamental in ensuring the transfer of our specialist knowledge to the
global community. [http://www.kew.org/]
Millennium Seed Bank Project, Royal Botanic Gardens Kew
The Millennium Seed Bank Project, which is managed by Kew’s Seed Conservation Department, is the largest
ex situ conservation project ever conceived. Its partners will have banked seed from 10% of the world's wild
plant species by the end of the decade. These will not be just any plants, but will include the rarest, most
threatened and most useful species known to man. Our scientific objectives are to increase knowledge and
understanding of seed diversity, particularly in relation to four key traits: seed maturity (development); seed
desiccation tolerance; seed storability; and seed dormancy (germination). These traits have direct relevance to
improving the seed conservation practices of seed 'collecting', 'processing', 'storing' and 'monitoring and using'.
The group's work is divided into seven themes including the ‘Science of Ageing’ and ‘Storage Technology’. The
research group has collaborations with about 50 countries. [http://www.kew.org/msbp/]
Society for Low Temperature Biology, SLTB
The Society for Low Temperature Biology (SLTB) was founded in 1964 and became a Registered Charity in
2003 (Charity Commission for England & Wales No. 1099747) with the purpose of promoting research into the
effects of low temperatures on all types of organisms and their constituent cells, tissues and organs. Such studies
have applications in a diverse variety of scientific fields from biology and medicine to engineering. Research
interests of members range from natural mechanisms of cold tolerance, cryopreservation of cells and tissues for
medical, agricultural and conservation purposes, low temperature microscopy, through to the physics and
physical chemistry of water and ice, and heat and mass transfer in biological systems. This wide range of
scientific and technical backgrounds facilitates cross-fertilization of ideas at our meetings.
The Society has members from a wide variety of scientific backgrounds dealing with diverse organisms,
materials and technologies. Areas of activity amongst the membership include:
 Cryopreservation of cells, tissues, organs and materials for medical purposes
 Cryopreservation of plant cells and tissues and cultures of algae, protozoa and filamentous fungi for the
purposes of sustainable agriculture, biotechnology and conservation of biodiversity
 Refrigeration of biological materials
 Preservation of human and animal gametes and embryos
 Tissue engineering
 Cold tolerance of natural biological systems including plants, invertebrates, animals and
microorganisms
 Low temperature aspects of hibernation
 Biochemistry of natural cryoprotectants and cryoprotection
 Cryomicroscopy
 Physics of supercooling, ice nucleation/formation and heat transfer
CryoLetters
CryoLetters is a bimonthly international journal for low temperature sciences, including cryobiology,
cryopreservation or vitrification of cells and tissues, chemical and physical aspects of freezing and drying, and
studies involving ecology of cold environments, and cold adaptation
The journal publishes original research reports, authoritative reviews, technical developments and commissioned
book reviews of studies of the effects produced by low temperatures on a wide variety of scientific and technical
processes, or those involving low temperature techniques in the investigation of physical, chemical, biological
and ecological problems.
Editorial Office: editor@cryoletters.org
CryoLetters
c/o LIRANS Institute of Research in the Applied Natural Sciences
University of Bedfordshire, 250 Butterfield, Great Marlings, Luton, Bedfordshire, LU2 8DL, UK
Tel: 44 (0)1582 743729. Executive Editor: Professor Barry Fuller
4
Fundamental Aspects of Plant Cryopreservation
17-18th February 2009
Royal Botanic Gardens, Kew, Wakehurst Place, West Sussex, UK
PROGRAMME:
Tuesday 17 Feb. 2009
13.30 h
Coach Departs Hotel
Meet at Hotel Lobby
14.30 –15:45 h Core group meeting
Wellcome Trust Millennium Building (WTMB)
Seminar Room
15:45-16.00 h
Refreshments
16:00-17:15 h
Sub-group meeting on
future funding and strategy
WTMB Seminar Room
17.30 h
Coach Departs Wakehurst
Place
WTMB
Wednesday 18 Feb. 2009
08.00 h
Coach Departs Hotel
Meet at Hotel Lobby
08.30-9.00 h
Registration
Outside WTMB Seminar Room
09.00-9.15 h
Welcome and introduction: Hugh W. Pritchard (UK)
SCIENCE MEETING SESSION 1
Chair : Dr Joachim Keller
09.15-9.45 h
Invited speaker
Erica Benson (UK): Exploring cryopreservation stress,
tolerance and recalcitrance in nature’s laboratory
09.45-10.00 h
Elke Heine-Dobbennack, Heiko Kiesecker and Heinz
Martin Schumacher (Germany): Fundamental aspects and
economic needs of plant cell cryopreservation
10.00-10.15 h
Thomas Roach , Beckett RP, Minibayeva F and Kranner I
(UK): Reactive oxygen species production from the
initial stages of cryopreserving Castanea sativa embryonic
axes
10.15-10.30 h
Sebastien C. Carpentier, Yves Lambeens and Bart Panis.
(Belgium): Unravelling sugar preculture: new nutrients,
excision and osmotic stress
10.30-11.00 h
Refreshments
Posters
11.00-11.15 h
Paul Lynch, M Al Majathoub & K Harding (UK):
Cryopreservation of in vitro shoot tips of Fragaria x ananassa
Duch.
11.15-11.30 h
Rachael Davies, J Nadarajan & H W Pritchard. (UK):
Cryopreservation of difficult-to-handle palm seeds
5
11.30-11.45 h
Matus Skyba and Eva Cellarova (Italy): Fundamental
Aspects in Cryopreservation of Medicinal Plants: The
Hypericum story.
11.45-12.00 h
Jiri Zamecnik and Milos Faltus (Czech Republic):
Evaluation of thermograms from Differential Scanning
Calorimeter.
12.00-12.15 h
Discussion
12.15-14.00 h
Lunch and tour of
WTMB including
Millennium Seed
Bank
Stable Restaurant and WTMB
SCIENCE MEETING SESSION 2
Chair: Professor Pawel Pukacki
14.00-14.30 h
Invited Speaker
Roger Pearce (UK): Complexity and a regulatory role for sugars in
the acclimation to cold and freezing of plants
14.30-14.45 h
Milos Faltus and Jiri Zamecnik (Czech Republic): Thermal
characteristics of some vitrification solutions
14.45-15.00 h
Brian Grout (Denmark): Practical benefits of dormant bud
cryopreservation for genetic resource conservation
15.00-15.15 h
Keith Harding, P T Lynch & J W Johnston (UK): Epigenetic
Changes Associated with the Cryopreservation of Clonal Crops
15.15-15.45 h
Refreshments
Posters
15.45-16.00 h
Elena L Peredo, M Ángeles Revilla, Borja Jiménez-Alfaro,
Álvaro Bueno, Eduardo Cires, J Antonio Fernández Prieto &
Richard J Abbott (Spain). Applications of molecular markers prior
conservation actions on the Spanish endemic Senecio boissieri
16.00-16.15 h
A. Gallard, J Escoute, J-L Verdeil and A Grapin (France)
Dynamic study of cellular events during cryopreservation
16.15-16.30 h
Aida Reis, Rena Farias, Ana Barata & Eugénia Nunes (Portugal):
The Conservation of Vegetatively Propagated Allium Collection in
the Portuguese Genebank
16.30-16.45 h
E. R. Joachim Keller (Germany): Chances for utilization of
cryopreservation in complex botanical collections – the
Alliaceae/Amaryllidaceae clade
16.45-17.15 h
General Discussion
17.30 h
Coach departs
WTMB
6
for Hotel
7
8
LIST OF PARTICIPANTS
Name
Address
Agnes GRAPIN
AGROCAMPUS-OUEST, INHP Angers, 2 rue Le Nôtre, 49045 Angers - F.,
France. Email: agnes.grapin@agrocampus-ouest.fr
Aida REIS
Banco Português de Germoplasma Vegetal, Unidade de Recursos Genéticos,
Ecofisiologia e Melhoramento de Plantas, INRB, I.P., Quinta de S. José, S. Pedro
de Merelim, 4700-859 Braga.Tel. 253 300 960 Fax. 253 300 961.
Email: areis@drapn.min-agricultura.pt
Alex WOODCRAFT
University of Sussex. Email: Albycraft@aol.com
Bart PANIS
Laboratory of Tropical Crop improvement, KULeuven, Kasteelpark Arenberg 13,
3001 Leuven, Belgium. Email: Bart.panis@biw.kuleuven.be
Brian GROUT
Faculty of Life Sciences, University of Copenhagen, Hoejbakkegaard Alle 21,
2630 Taastrup, DENMARK. Email: bwg@life.ku.dk
Carla BENELLI
Istituto per la Valorizzazione del Legno e delle Specie Arboree, Consiglio
Nazionale delle Ricerche,Via Madonna del Piano, 10 - 50019 Sesto Fiorentino
(FI)- Italy. Email: benelli@ivalsa.cnr.it
Carmen MARTIN
Dpto. de Biología Vegetal, E.U.I.T. Agrícola, Universidad Politécnica de Madrid,
Ciudad Universitaria, 28040 Madrid, Spain. Email: mariacarmen.martin@upm.es
Elena GONZÁLEZ-BENITO
Dpto. de Biología Vegetal, E.U.I.T. Agrícola, Universidad Politécnica de Madrid,
Ciudad Universitaria, 28040 Madrid, Spain. Email: me.gonzalezbenito@upm.es
Elena LÓPEZ PEREDO
Universidad de Oviedo, Deparment of Plant Physiology, Catedrático Rodrigo Uría
s/n, 33071 Oviedo, Spain. Email: elperedo.uo@uniovi.es
Emilia CABONI
CRA - Fruit Tree Research Center, Via di Fioranello, 52, 00134 - Rome Italy.
Email: e.caboni@propag.org
Erica BENSON
Damar Research Scientists, Conservation, Environmental Science &
Biotechnology, Damar, Drum Road, Cuparmuir, Fife, KY15 5RJ, Scotland, UK.
Email: e.benson-damar@tiscali.co.uk
Eva ČELLÁROVÁ
Institute of Biology and Ecology, Faculty of Science,P. J. Šafárik University,
Mánesova 23, 04154 Košice, Slovakia. Email: eva.cellarova@upjs.sk
Hugh W. PRITCHARD
Seed Conservation Department, Royal Botanic Gardens Kew, Wakehurst Place,
Ardingly, West Sussex RH17 6TN, UK. Email: h.pritchard@kew.org
Jan RYBCZYŃSKI
Botanical Garden-Center for Biological Diversity Conservation, Polish Academy
of Sciences, ul. Prawdziwka 2, o2-973 Warsaw, POLAND.
Email: jjryb@neostrada.pl
Jayanthi NADARAJAN
Seed Conservation Department, Royal Botanic Gardens Kew, Wakehurst Place,
Ardingly, West Sussex RH17 6TN, UK. Email: j.nadarajan@kew.org
9
Jiri ZAMECNIK
Crop Research Institute, Drnovska 507, CZ 161 06 00 prague, Czech Republic.
Email: zamecnik@vurv.cz
Joachim KELLER
Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3,
D-06466 Gatersleben, Germany. Email: keller@ipk-gatersleben.de
Jon GREEN
Genómica Funcional, CIC-Biogune, Parque Tecnológico de Vizcaya, Edificio
801A, 1pl. 48160-Derio, Spain. Email: jgreen@cicbiogune.es
Keith HARDING
Damar Research Scientists, Conservation, Environmental Science &
Biotechnology, Damar, Drum Road, Cuparmuir, Fife, KY15 5RJ, Scotland, UK
Email: k.harding-damar@tiscali.co.uk
Martin SCHUMACHER
DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH,
Inhoffenstraße 7B, 38124 Braunschweig, Germany. Email: mas@dsmz.de
Milos FALTUS
Crop Research Institute, Drnovska 507, CZ 161 06 00 prague, Czech Republic
Email: faltus@vurv.cz
Paul LYNCH
Biological Sciences Research Group, University of Derby, Kedleston Road Derby,
DE22 1GB UK. Email: p.t.lynch@derby.ac.uk
Pawel PUKACKI
Physiol of Abiot Stress Lab., Inst of Dendrology Polish Academy of Sci, PL-62035 Kornik, POLAND. Email: ppukacki@man.poznan.pl
Rachael DAVIES
Seed Conservation Department, Royal Botanic Gardens Kew, Wakehurst Place,
Ardingly, West Sussex RH17 6TN, UK. Email: rachael.davies@kew.org
Roger PEARCE
School of Biology, Ridley Building, Newcastle University, Newcastle upon Tyne
NE1 7RU. Email: r.s.pearce@ncl.ac.uk
Sebastien CARPENTIER
Laboratory of Tropical Crop improvement, KULeuven, Kasteelpark Arenberg 13,
3001 Leuven, Belgium. Email: Sebastien.carpentier@biw.kuleuven.be
Thomas ROACH
Seed Conservation Department, Royal Botanic Gardens Kew, Wakehurst Place,
Ardingly, West Sussex RH17 6TN, UK. Email: t.roach@kew.org
ATTENDEES FROM KEW GARDENS
2 Staff from Conservation Biotechnology Unit, RBG Kew, London
4 staff from Curation Section, Seed Conservation Dept
3 staff / students from Research Section, SCD
2 staff / students from Technology and Training Section, SCD
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NOTES:
11