What is CRYMCEPT ??

1. QLK5-CT-2002-012791 November, 2002- 31 October, 2005 What is CRYMCEPT ??

2. 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 (embryogenic cell suspensions) • Eradication of viruses (Banana, Plum) • Conservation of plant pathogens (fungi, nematodes)

3. 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 • So no real standardised methods are available • 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). • A more efficient optimisation of existing cryopreservation protocols and/or development of new protocols is only possible if the physico-chemical background of cryoprotection/tolerance towards dehydration is known.

4. Objectives General objective: The development of new/improved cryopreservation protocols applicable to plant germplasm collections in Europe. • Specific objectives: • Development of sensitive, reliable and use friendly techniques to measure plant species / tissue response to cryoprotection/cryopreservation. • Determination of physico-biochemical changes associated with tolerance towards cryopreservation. • Development of new plant cryopreservation protocols. • Validation of the improved cryopreservation protocols and their application to European germplasm collections.

5. Physico-biochemical parameters related to plant cryopreservation Proteins Sugars Water thermal behavior Polyamines Membrane components Cytoskeletal protein Oxidative stress Plant cell

6. Project workplan Work packages (WP) Water thermal behaviour (WP1) Protein (WP2) Sugar (WP3) Membrane components (WP4) Cryopreservation protocols Polyamines (WP5) Cytoskeletal protein (WP6) Oxidative stress (WP7) Development of new/adapted plant cryopreservation protocols (WP8) Validation of the improved cryopreservation protocols and their application/dissemination to European germplasm collections (WP9)

7. Plant species 1. Banana Non-hardening Monocot 2. Coffee Non-hardening Dicot 3. Garlic Hardening Monocot 4. Potato Hardening Dicot 5. Ribes Hardening Dicot 6. Olive Hardening Dicot 7. Apple Hardening Dicot 8. Almond Hardening Dicot

8. Partners Considerable experience with plant cryopreservation preferentially linked with a germplasm collectionExperience in and/or interest for the analysis techniques that are described in the different Work packages. Partner 1. K.U.Leuven Catholic University of Leuven, Lab. of Tropical Crop Improvement, Belgium in collaboration with Catholic University, Lab. of Plant Physiol. Belgium Partner 2. UAD University of Abertay Dundee, School of Molecular Life Science, U.K. Partner 3. UNIV1 University of Derby, Division of Biological Science, U.K. Subcontractor: Nottingham Trent University, U.K. Partner 4. IRD Institut de Recherche pour le Développement, GeneTrop - Génétique des Plantes, France Partner 5. ISF - Rome Fruit Tree Reserch Institute, Propagation Department, Italy Subcontractor: University of Tor Vergata, Italy Partner 6. DSMZ Deutsche Sammlung Mikroorganismen Zelkulkulturen GmbH, Plant Cell Depart., Germany Partner 7. IPGRI International Plant Genetic Resources Institute, Italy

9. Work packages Partners Plant species WP1 P1 1. Banana WP2 2. Ribes P2 WP3 3. Garlic P3 WP4 4. Olive WP5 P4 5. Coffee WP6 P5 6. Apple WP7 P6 7. Almond WP8 8. Potato WP9 P7

10. Project management and coordination Project coordinator HES 1 K. U. Leuven Belgium Steering group (Co-ordinator + Partner’s representatives) OTH 7 IPGRI Italy Collaborator Lab Plant Physiol. K. U. Leuven REC 4 IRD France HES 3 Univ Derby UK HES 2 Univ Ab-Dun UK REC 5 ISF Italy REC 6 DSMZ Germany Subcontractor Univ Nottingham Trent Subcontractor Univ Tor Vergata

11. How to maximize chance for success ? • Each Work package involves at least 2 partners. The Work package coordinator has already proven to have relevant and considerable experience with the analysis technique that is described in the Work package. • All partners 1- 6 are on average participating actively in 3 to 4 of the Work packages 1-7. This ensures, together with the yearly coordination meetings an efficient communication flow between the Work package groups. • The project covers a variety of the most relevant physico-biochemical parameters that could be related to cryoprotection. Nevertheless, at each yearly coordination meeting a discussion will be held to ensure that key issues will not fall between WPs. • The project involves a very broad spectrum of genotypes, species (monocots and dicots), different tissues (differentiated and dedifferentiated) with a differential response towards hardening (hardening not possible to complete hardening). As such, a series of generally applicable “core” cryopreservation will be established. These will be applicable to germplasm which display different sensitivities and tolerances (e.g. cold tolerant, desiccation sensitive)

12. How to maximize chance for success ? 20-21 Feb 2003 11-13 Dec 2002 12-14 Nov 2003

13. September 13, 2003

14. September 13, 2003

15. Website: http://www.agr.kuleuven.ac.be/dtp/tro/CRYMCEPT/