Transcript Slide 1

COST Technical Committee "Agriculture, Biotechnology and Food Science"
Proposal for a new COST Action
Cryopreservation of crop species in Europe
ACRONYM: CRYOPLANET
Proposing country: Belgium
Action Proposer : Bart Panis
Plant Genetic Resources: Current situation
Europe
•64 endemic plants of Europe have become extinct in recent decades
•24% of the species/subspecies of certain groups of European plants
are in danger to be lost
•Agricultural intensification has reduced the area under wetlands in
Europe by some 60 % in the last decades with consequent
threats on biodiversity
Worldwide
•100,000 plants representing 1/3rd of plant species are threatened
•Since the 1970s large number of land races and wild relatives are
sampled and stored ex situ
•Now, about 6 million samples are held in national, regional, international
and private genebank collections
Storage of Plant germplasm
• In situ : Conservation in ‘normal’ habitat
–rain forests, gardens, farms
• Ex Situ :
–Field collection, Botanical gardens
–Seed collections
–In vitro collection
•Normal growth
•Slow growth (temp, O2 , H2O , medium ~)
•Cryopreservation (-196°C)
• (DNA Banks)
What is cryopreservation ?
•What ?
–Storage of living tissues at ultra-low temperatures (-196°C)
•Use
–Conservation of plant germplasm
•Vegetatively propagated species (root and tubers, ornamental, fruit
trees)
•Recalcitrant seed species (Howea, coconut, coffee)
–Conservation of tissue with specific characteristics
•Medicinal and alcohol producing cell lines
•Genetically transformed tissues
•Transformation/Mutagenesis competent tissues (ECSs)
–Eradication of viruses (Banana, Plum)
–Conservation of plant pathogens (fungi, nematodes)
What is the ‘problem’ with plant cryopreservation ?
•Cryopreservation procedures are now available for about 150200 different plant species
• But for each species and tissue type, the cryopreservation
protocol needs to be empirically adapted in function of their
• natural freezing resistance
• explant size and type
• water content
• Most of the work on cryopreservation of plants has been
performed in the framework of academic studies and involves
only one or a few genotypes. Only few plant germplasm
collections stored in liquid nitrogen currently exist (with a relatively
limited amount of accessions).
No real standardised methods are available !!!
Examples of existing collections in liquid nitrogen?
•National Seed Storage Laboratory (NSSL) (Fort Collins, Colorado, USA):
2,100 accessions of apple (dormant buds)
•National Clonal Germplasm Repository (NCGR) of Corvallis (USA): 104
accessions of pear (shoot tips);
•International Potato Centre (CIP) (Lima, Peru) : 345 potato accessions
•Tissue Culture BC Research Inc.(Vancouver, BC, Canada) : 5000 accessions
representing 14 conifer species
•AFOCEL (Association Forêt Cellulose) of France, with over 100 accessions of
elm (dormant buds);
•National Institute of Agrobiological Resources (NIAR) of Japan, with about
50 accessions of mulberry.
•IRD (Montpellier, France) : 80 accessions of oil palm
•German Collection of Micro-organisms and Cell Cultures (DSMZ)
(Braunschweig, Germany) : 519 old potato varieties
•INIBAP, Laboratory of Tropical Crop Improvement, K.U.Leuven (Heverlee,
Belgium) : 440 banana accessions
Why is cryopreservation not more widely applied?
(i)
the unavailability of efficient and robust cryopreservation protocols
applicable to many plant species and diverse germplasm types
(ii) limited awareness of plant researchers unacquainted to recent
developments in cryogenic storage methods
(iii) lack of coordinated research on plant cryopreservation.
 COST action like “CRYOPLANET” could make the difference
Why are the objectives of CRYOPLANET ?
Objective 1: To screen in detail the current utilization of plant
cryopreservation in Europe.
Objective 2: To screen and compare the efficiency of existing plant
cryopreservation protocols.
Objective 3: To improve fundamental knowledge about
cryoprotection through the determination of physicobiochemical changes associated with tolerance towards
cryopreservation.
Objective 4: To develop new plant cryopreservation protocols.
Objective 5: To assure the genetic stability and true-to-typeness of
plants after cryopreservation.
Objective 6: To apply cryopreservation to European plant germplasm
collections.
Objective 7: To proof the environmental, social and economic impact
of plant cryopreservation.
Scientific programme of CRYOPLANET ?
WG1
Fundamental aspects of
cryopreservation/cryoprotection and
genetic stability
Optimisation
Feed back
WG2
Technology, application and validation
of plant cryopreservation
WG1: Fundamental aspects of cryopreservation/cryoprotection
and genetic stability
1.1. Fundamental aspect of cryopreservation and cryoprotection
Elucidation of the physico-biochemical background of cryoprotection and
cryopreservation.
Water thermal
behavior
Proteins
Sugars
Membrane
components
Polyamines
Cytoskeletal
protein
Oxidative
stress
WG1: Fundamental aspects of cryopreservation/cryoprotection
and genetic stability
1.2. Genetic stability and authenticity
Assessment of the genetic integrity of plants to determine if they are ‘true to type’ after
cryopreservation.
• Assessments of phenotypic variation (morphological descriptors)
• Cytological techniques to detect various types of chromosomal instability.
(polyploidy, aneuploidy and other mitotic abnormalities)
• Biochemical metabolite/protein (isozyme) profiles
• Genomic DNA sequences that can be analysed using a range of hybridization and
(PCR) techniques
• Epi-genetic variation in chromatin and DNA methylation of gene sequences
WG2:Technology, application and validation of plant
cryopreservation
2.1.Technology aspects of cryopreservation
Applications of different cryopreservation protocols to different plant species and
tissues.
WG2:Technology, application and validation of plant
cryopreservation
2.2. Impact and applications of cryopreservation in plants
genebanks, establishment of cryo-bank and dissemination of results
Organisation of CRYOPLANET
The Management Committee (MC)
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Appointment of Action Chair, Vice-Chair(s) and WG Co-ordinators.
Planning of MC meetings and of Scientific Meetings and Workshops.
Assessment and report of the progress made by the different WGs
Promotion of co-operation and of data exchange between the WGs.
Promotion and approval of Short-Term Scientific Missions,
Preparation of the Annual Reports.
Establishment and update of a Web site
Organization of contacts and common workshops
Preparation of a EU 7th framework project
Organisation of CRYOPLANET
Two Working Groups
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WG1 : Fundamental aspects of cryopreservation/cryoprotection and
genetic stability
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WG2: Technology, application and validation of plant cryopreservation
Working Group meetings will be organised on a yearly base
WG coordinators
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Planning the appropriate Scientific Meetings.
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Coordination of the activities within the WG.
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Promoting joint research (f.e. STSMs) and common publications.
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Report on the WG progress to the Action Chair and MC
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Participation in the plenary and restricted MC meetings.
Inter-COST Workshops
Short-Term Scientific Missions (STSM),
Time table
Year 1
Year 2
Year 3
Year 4
Coordination
Kick-off
meeting
Homepage
Reporting
MC meeting
WG1 meeting
WG2 meeting
Workshop*
STSMs
Final Conference
MC meeting: Management committee meeting; WG meeting: Working group meeting; STSMs: Shortterm scientific missions; Workshop*: Timing of the Inter-COST Workshops will be defined in agreement
with the Management committee of that specific Action.
Economic dimensions
•17 COST countries
•64 researchers
•50 research institutes
Dissemination
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Articles in refereed scientific journals
Common reviews, books
A public website (information about the project, the achievements,
services/consultancies offered and announcements of training workshops.
Information on the official webpages of the collaborating
institutions.
The consortium will organize workshops for scientists, germplasm
curators regulatory bodies and policy makers
At the end of the Action, the consortium will offer its expertise as a
service to the EU.
Presentations at International Conferences, for promoting the
European know-how and increasing the international collaboration.
Teaching activities in Universities at undergraduate and post-graduate
level.Young scientists and engineers will thus be trained and informed on
the latest developments in cryopreservation.
Acknowledgements
Carpentier Sebastien, Druart Philippe, Geelen Danny, Swennen Rony, Tsvetkov
Ivaylo, Bilavcik Alois, Faltus Milos, Zamecnik Jiri, Antonius-Klemola Kristiina,
Häggman Hely, Nukari Anna, Rokka Veli-Matti, Uosukainen Marjatta, Dussert
Stéphane, Engelmann Florent, Grapin Agnès, Harvengt Luc, Malaurie Bernard,
Péros Jean-Pierre, Trontin Jean-François, Höfer Monika, Keller Joachim, MeierDinkel Andreas, Pinker Ina, Schumacher Heinz Martin,Benelli Carla, Caboni
Emilia, Damiano Carmine, Dulloo Ehsan, Lambardi Maurizio, Hausman JeanFrançois, Criel Bram, Harrouni Cherif, de Klerk Geert-Jan, Raemakers Krit, Bach
Anna, Lisek Anna, Mikuła Anna, Niedzielski Maciej, Pukacki Pawel M., Rybczyński
Jan J., Sochacki Dariusz, Zimny Janusz, Miguel Célia, Reis Moura Isabel,
Oliveira¨Margarida,Halmagyi Adela, Palada Magdalena, Rakosy-Tican Elena,
Gavrilenko Tatjana, Salaj Terezia, González-Benito Elena, Revilla M. Ángeles,
Vieitez Ana, Pâques Marc, Feki Lotfi, Aylin Ozudogru Elif, Ozden-Tokatli Yelda,
Benson Erica, Cripps Ryan, Grout Brian, Harding Keith, Lynch Paul T., Pritchard
Hugh W.,Wetten Andy