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

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COST Technical Committee "Agriculture,
Biotechnology and Food Science"Proposal for a
new COST ActionCryopreservation of crop species
in EuropeACRONYM CRYOPLANETProposing
country BelgiumAction Proposer Bart Panis
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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

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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 (-196C)
• (DNA Banks)

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What is cryopreservation ?

• What ?
• Storage of living tissues at ultra-low
  temperatures (-196C)
• 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)

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What is the problem with plant cryopreservation
?

• Cryopreservation procedures are now available for
  about 150-200 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 !!!

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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

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Why is cryopreservation not more widely applied?

• 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.
• ? COST action like CRYOPLANET could make the
  difference

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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 physico-biochemical 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.
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Scientific programme of CRYOPLANET ?

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WG1 Fundamental aspects of cryopreservation/cryop
rotection and genetic stability
1.1. Fundamental aspect of cryopreservation and
cryoprotection Elucidation of the
physico-biochemical background of cryoprotection
and cryopreservation.
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WG1 Fundamental aspects of cryopreservation/cryop
rotection 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

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WG2 Technology, application and validation of
plant cryopreservation
2.1. Technology aspects of cryopreservation Applic
ations of different cryopreservation protocols to
different plant species and tissues.
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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

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Organisation of CRYOPLANET

• The Management Committee (MC)
• 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

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Organisation of CRYOPLANET

• Two Working Groups
• WG1 Fundamental aspects of cryopreservation/cryo
  protection and genetic stability
• WG2 Technology, application and validation of
  plant cryopreservation
• Working Group meetings will be organised on a
  yearly base
• WG coordinators
• Planning the appropriate Scientific Meetings.
• Coordination of the activities within the WG.
• Promoting joint research (f.e. STSMs) and common
  publications.
• Report on the WG progress to the Action Chair and
  MC
• Participation in the plenary and restricted MC
  meetings.
• Inter-COST Workshops
• Short-Term Scientific Missions (STSM),

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Time table
Year 1 Year 1 Year 1 Year 1 Year 2 Year 2 Year 2 Year 2 Year 3 Year 3 Year 3 Year 3 Year 4 Year 4 Year 4 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 Short-term
scientific missions Workshop Timing of the
Inter-COST Workshops will be defined in agreement
with the Management committee of that specific
Action.
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Economic dimensions

• 17 COST countries
• 64 researchers
• 50 research institutes

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Dissemination

• 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.

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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, Meier-Dinkel Andreas, Pinker Ina,
Schumacher Heinz Martin,Benelli Carla, Caboni
Emilia, Damiano Carmine, Dulloo Ehsan, Lambardi
Maurizio, Hausman Jean-François, Criel Bram,
Harrouni Cherif, de Klerk Geert-Jan, Raemakers
Krit, Bach Anna, Lisek Anna, Mikula Anna,
Niedzielski Maciej, Pukacki Pawel M., Rybczynski
Jan J., Sochacki Dariusz, Zimny Janusz, Miguel
Célia, Reis Moura Isabel, OliveiraMargarida,Halma
gyi 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