COST Technical Committee "Agriculture, Biotechnology and Food Science"

Proposal for a new COST Action

Cryopreservation of crop species in Europe

ACRONYM: CRYOPLANET

Proposing country: BelgiumAction Proposer : Bart Panis

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

Plant Genetic Resources: Current situation

• 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)

Storage of Plant germplasm

•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 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 !!!

What is the ‘problem’ with plant cryopreservation ?

•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

Examples of existing collections in liquid nitrogen?

(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 is cryopreservation not more widely applied?

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.

Why are the objectives of CRYOPLANET ?

Scientific programme of CRYOPLANET ?

WG1Fundamental aspects of

cryopreservation/cryoprotection and genetic stability

WG2Technology, application and

validation of plant cryopreservation

Optimisation Feed back

1.1. Fundamental aspect of cryopreservation and cryoprotection

Elucidation of the physico-biochemical background of cryoprotection and cryopreservation.

WG1: Fundamental aspects of cryopreservation/cryoprotection and genetic stability

Water thermalbehavior

Cytoskeletalprotein

Membranecomponents

Proteins Sugars

Polyamines

Oxidativestress

1.2. Genetic stability and authenticity Assessment of the genetic integrity of plants to determine if

they are ‘true to type’ after cryopreservation.

WG1: Fundamental aspects of cryopreservation/cryoprotection and genetic stability

• 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

2.1. Technology aspects of cryopreservationApplications 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

WG2: Technology, application and validation of plant cryopreservation

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

Organisation of CRYOPLANET

Two Working Groups• WG1 : Fundamental aspects of

cryopreservation/cryoprotection 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),

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: Short-term 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

• 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, 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, 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