Task 1 : Acquisition of Plant Materials and Data

GÉNÉRAL OBJECTIVES

The main objective of GrapeGen06 is to contribute to the successful long term preservation of the Vitis genetic resources for the use of the future generations. By ensuring an optimized characterization of these resources, we will promote their use and ensure the competitiveness and robustness of the production sector. The following goals are planned: 1. Monitor still existing Vitis germplasm in Europe, including Eastern countries, 2. Describe this germplasm, using common morphological, agronomic and molecular descriptors and linking them to a unique web-database (http://www.genres.de/eccdb/vitis) 3. Implement true-to-type identification of varieties decreasing the number of errors, homonyms and synonyms affecting the European collections. 4. Promote an overall optimized conservation scheme for the safe conservation of the Vitis germplasm including Vitis sylvestris presently threatened of extinction, and involving several conservation means (ex-situ collections, cryopreservation and on-farm conservation by farmers and professionals) so that the resources are conserved, accessible and field-tested in a pertinent agricultural context.

PROGRAMME DIVIDED INTO TWO MAIN PARTS :

TASK 1 : ACQUISITION OF PLANT MATERIALS AND DATA

TASK 2 : CONSERVATION AND MANAGEMENT OF EUROPEAN GENETIC RESOURCES

WP1 -. Data acquisition for the SSR-marker database within the EU-Vitis database: WP2. Acquisition of primary and secondary descriptor data of old, endangered and autochthonous grape varieties WP 3. On-farm evaluation of agronomic features of autochthonous grape varieties WP 4. Study, identification and evaluation of Vitis sylvestris germplasm WP 5. True-to-type analysis WP 6 Database improvement and enrichment WP 7 Elaboration of a long term conservation strategy (integrating several types of conservation, in-situ, ex-situ, cryoconservation) WP1 -. Data acquisition for the SSR-marker database within the EU-Vitis database: Global objective of WP1 : molecular analysis of genetic resources in order to resolve true-to-type identification and for resolution of problems of misnaming, synonyms and homonyms Objective 1 : Definition of SSR markers suitable for identification purpose, and definition of the references alleles for coding Objective 2 : Characterization of grapevine germplasm using these defined SSR markers Objective 3: Dissemination of data Task 1 : Encoding of already available data for the 6 SSR from Genres Task 2 : Selection of at least 6 new markers and definition of reference alleles Task 3: Microsatellite analysis of chosen accessions and encoding Task 4: Coordinated microsatellite analysis as needed within WP5 Description of work, milestones and deliverables per year Year 1  6 Genres081 SSR-markers: collection of available data from partners and publications, which are suited for coding. Coding, compilation and comparison of data  Additional high polymorphic SSR markers:. o all partners will work with identical DNA (from one accession) which will be distributed either through shipment of DNA or through shipment of wood, o the same 45 example varieties as in Genres081 will be analysed for procedure harmonisation M1 : April 2007 Definition of the list of SSR markers and cultivars to analyse D1 : April 2007 List of cultivars to be analyzed D2: April 2007 List of SSR-markers necessary for analysis D3: October 2007 Software for automatic coding Year 2  6 Genres081 SSR-markers: in the case of divergent SSR-marker data results, each participating laboratory will verify the problematic accessions (max. 20). In the EU-Vitis database the verified data will be published and linked to the accessions. A specific SSR- marker data search catalogue will be established (WP6). Collection, coding and comparison of further SSR-marker data will continue.  Additional high polymorphic SSR markers: Extraction of DNA, distribution and analysis of 45 varieties / accessions. : compilation and comparison of allele lengths , selection of reference alleles able to discriminate all the available varieties, definition of OIV descriptors for the 6 STMS-markers M2 : January 2008 Choice of the cultivars and comparison of data between laboratories for reference alleles D4 : January 2008 List of reference cultivars and alleles for new cultivars M3 : January 2008 Coding of SSR data and transfer to WP6 (for available data) Year 3  6 Genres081 SSR-markers: the same as in year 2.  Additional high polymorphic SSR markers: Collection, coding and comparison of SSR-marker data for the cultivars already analyzed with the other markers M4: January 2009 Coding of SSR data and transfer to WP6 (for available data) Year 4  6 Genres081 SSR-markers: the same as in year 2  Additional high polymorphic SSR markers: Collection, coding and comparison of SSR-marker data , Shipment of consistent data to coordinator WP6. M5: January 2010 Coding of SSR data and transfer to WP6 (for available data) M6: October 2010 Coding of SSR data and transfer to WP6 (for available data) M7: December 2010 Publication of complete data in the database WP2. Acquisition of primary and secondary descriptor data of old, endangered and autochthonous grape varieties

Global objective of WP2 : Analysis of autochthonous varieties for primary and secondary descriptors and for sanitary status. Main emphasis is laid on germplasm of Eastern European countries. Objective 1 : Standardized description of old autochthonous varieties with special emphasis on germplasm of Eastern European countries Objective 2 : Analysis of sanitary status of autochthonous varieties Objective 3: Dissemination of data Task 1 : Recording of 32 primary and secondary descriptors analogous to Genres CT96 No81. Utilisation of the descriptors recently harmonized by UPOV, IPGRI and OIV Task 2 : Realisation of the pictures of the cultivars Task 3: Characterization of sanitary status of the cultivars Task 4 : Preparation of the catalogue of autochthonous cultivars

Description of work, milestones and deliverables per year Year 1  Harmonisation and centralisation of existing data available with the participants and their compilation into the database.  Discussion and decision concerning the inclusion of phytopathology tests (viroses), which is a necessary requirement to produce, exchange and distribute safe materials.  Recording of primary and secondary descriptors of old, endangered and autochtonous varieties  Realisation of pictures M8 : April 2007 Harmonisation and centralization of already existing data available D6: April 2007 List of a new set of analysis for phytosanitary test (viruses). D7: April 2007 List of old, endangered and autochthonous varieties to be analysed. Year 2 and year 3  Recording of primary and secondary descriptors of old, endangered and autochtonous varieties / accessions in collections..  Phytosanitary tests (viruses)  Data transfer to the coordinator  Realisation of pictures  Data transfer to WP6 M9 : January 2008 Shipment of Primary, secondary descriptors data and pictures data D8: January 2009 Primary, secondary descriptors data and pictures already recorded M10: January 2009 Shipment of Primary, secondary descriptors data and pictures data to the coordinator Year 4  Recording of primary and secondary descriptors of old, endangered and autochtonous varieties / accessions in collections..  Phytosanitary tests (viruses)  Realisation of pictures  Data transfer to WP6 M11: October 2010 Shipment of Primary, secondary descriptors data and pictures data to the coordinator M12 : December 2010 Publication of the descriptor data via the European Vitis database. M13: December 2010 Catalogue of autochthonous and endangered varieties in Europe. D9: December 2010 Primary and secondary descriptors data and pictures available via the EU-Vitis-database D5: publication of all data in the database WP 3. On-farm evaluation of agronomic features of autochthonous grape varieties Global objective of WP3 : develop a knowledge on some of the autochthonous varieties to be used by growers in EU based on field data. Objective 1 : Improving our knowledge of agronomic and wine features of neglected varieties Objective 2 : Promoting their availability (materials and information) to growers Objective 3: Dissemination of data Task 1 : Identification and characterization of suitable old vineyards : (identification of sites, growers, cultivars, Verification of cultivar identity if needed (in relation w WP1 and WP2,….) Task 2: Evaluation of agronomic features & wine quality Task 3 : Evaluation of sanitary features and selection of agronomically-interesting virus-free grape varieties / accessions Task 4: Information to growers by publication of activities and outcome in wine growers magazines Description of work, milestones and deliverables per year Year 1  Old vineyards: localisation of suitable old vineyards planted with endangered grape varieties or outstanding clones of cultivars still in use, with sufficient number of plants to allow agronomic evaluation and wine quality studies. Data transfer to WP6 M14: April 2007 Definition of the protocols to be followed D10 : April 2007 Protocol for evaluation of the different characteristics D11 : April 2007 List of vineyards and the cultivars to be analyzed Year 2  Old vineyards: identification and inventory of grape varieties. SSR-marker analysis, at least for doubtful grape varieties. Recording of agronomic traits. Test on virus infestation.Data transfer to WP6 M15 : Jan. 2008 Inventory of vineyards suitable for analysis and registration of location characteristics Year 3  Old vineyards: Recording of agronomic traits. Test on virus infestation. Selection of virus free clones representing the natural diversity of the variety. Data transfer to WP6 M16: January 2009 Shipment of agronomic features, wine quality, virus infestation to the coordinator of WP6 D12: January 2009 Description of the agronomic features of the analyzed varieties / clones already recorded Year 4  Old vineyards: Recording of agronomic traits. Test on virus infestation. Selection of virus free clones representing the natural diversity of the variety. Registering of the variety and site data into the European Vitis-database. Data transfer to WP6 M17: January 2010 Shipment of agronomic features, wine quality, virus infestation to the coordinator of WP6 D13: October 2010 Description of the agronomic features of the analyzed varieties / clones (final data) M18: October 2010 Shipment of agronomic features, wine quality, virus infestation to the coordinator of WP6 M19: December 2010 Publication of all data in the database D14: December 2010 Description of the sanitary status of the analyzed varieties / clones D15: December 2010 Publication of the results in wine growers magazines to promote the use of rare varieties and outstanding clones of varieties which are still in use WP 4. Study, identification and evaluation of Vitis sylvestris germplasm Global objective of WP4 : Inventory and analysis of V. silvestris populations in Europe Objective 1 : Inventory of Vitis sylvestris populations in Europe Objective 2 : Morphological, agronomic and genetic characterization of material Objective 3: Dissemination of data (except exact location for safety concern) Task 1 : Inventory of material (including evaluation of preservation status) Task 2 : Evaluation of morphological and/or agronomic features Task 3: Characterization of Vitis silvestris gene pool by microsatellite analysis Description of work, milestones and deliverables per year Year 1  Determination of Vitis sylvestris sites , Inventory of the Vitis sylvestris vines at those sites using GPS, Preservation status evaluation.  Definition of vine sampling for morphological description by primary and secondary descriptors D16: April 2007 Protocol for the inventory of V. sylvestris Year 2  Determination of additional Vitis sylvestris sites  Morphological description by primary and secondary descriptors  Agronomical evaluation, recording the resistance to biotic and abiotic stress factors and agronomical features, on in-situ and ex-situ plants. D17: January 2008 First list of sites with Vitis sylvestris plants in Europe and status regarding their protection Year 3  Determination of additional Vitis sylvestris sites  Morphological description by primary and secondary descriptors  Agronomical evaluation, recording the resistance to biotic and abiotic stress factors and agronomical features, on in-situ and ex-situ plants  Characterization of Vitis sylvestris genepool by SSR-marker analysis  Collection of cuttings / seeds from wild vines with agronomical interest (WP7) M20 : Jan 2009 Inventory of sites in Europe D19: January 2009 Second list of sites with Vitis sylvestris plants in Europe and their status regarding their protection M 21: Jan 2009 Shipment of morphological and agronomical data to WP6 Year 4  Determination of additional Vitis sylvestris sites  Morphological description by primary and secondary descriptors  Agronomical evaluation, recording the resistance to biotic and abiotic stress factors and agronomical features, on in-situ and ex-situ plants  Characterization of Vitis sylvestris genepool by SSR-marker analysis. Assessment of the genetic distance to other European occurrences and to cultivated grapes  Collection of cuttings / seeds from wild vines with agronomical interest (WP7)  Data transfer to WP6 M22: Jan 2010 Shipment of morphological and agronomical data for the wild population to WP6 D20: January 2010 Description of the agronomic features of the analyzed populations M23 : October 2010 Shipment of SSR, morphological and agronomical data to WP6 M24: December 2010 Publication of protocol for identification of V. sylvestris D21: December2010 - Description of the agronomic features of the analyzed populations D22: December 2010 - Final list of of sites with Vitis sylvestris plants in Europe and their status regarding their protection D23: December 2010 - Protocol for identification of Vitis sylvestris in Europe D24: December 2010 - SSR data for the V. silvestris populations WP 5. True-to-type analysis Global objective of WP5 : Sorting the EU collection for identical materiel = Beginning of a joint effort towards the management, monitoring and thus safeguard of grapevine germplasm. Objective 1 : Sorting EU collection for synonyms, homonyms,… Objective 2 : Assessment of the trueness-to-type of accessions in grapevine collections Objective 3 : First attempt toward the EU catalogue of cultivars and genetic resources. Objective 4: Dissemination of data Task 1 : identification of questionable varieties (list and analysis of the cultivar by ampelography and/or SSR data. Task 2 : establishment of the catalogue of genetic resources held in EU by comparison of collection composition and search for synonyms or homonyms. Task 3: Publication of data Description of work, milestones and deliverables per year Year 1  Collecting suggestions from partners on questionable samples to be examined/analyzed during the project in the different countries/regions  Comparison of inconsistencies or synonyms between variety names on the base of the SSR data (WP 1) and verification through the ampelographic features (WP 2) when available M25: April 2007 Definition of the list of questionable cultivars to be verified in Year 1 Year 2 For dubious/questionable cases, re-evaluation of the original resources (ampelography characters, SSR data) and analysis of bibliography M26: Jan 2008 Shipment of data recorded for year 1 on questionable cultivars D25: January 2008 First list of cultivars with questionable status and their true status M27: April 2008 Definition of the list of questionable cultivars to be verified in Year 2 Year 3 For local, old, autochthonous varieties, analysis of bibliographic sources and comparison to the actual descriptions (from EU-Vitis database or obtained throught WP2) Comparison of inconsistencies or synonyms between variety names on the Citations of synonyms and homonyms reported into the database and literature Correcting cases of misnaming M28: Jan 2009 Shipment of data recorded for year 2 on questionable cultivars D26: January 2009 - Extended list of cultivars with questionable status and their true status M29: April 2009 Definition of the list of questionable cultivarsto be verified in Year 3 Year 4 Preparing a list of synonyms/misnames/well definited homomyns to be incorporated into the EU- Vitis database. Preparing a list of identified old autochthonous varieties M30: Jan 2010 Shipment of data recorded for year 3 on questionable cultivars D27: January 2010 - Extended list of cultivars with questionable status and their true status M31 : April 2010 Definition of the list of questionable cultivars to be verified in Year 3 M32 : Oct 2010 Shipment of data recorded for year 3 on questionable cultivars D28: October 2010- List of synonyms, proper definition of homonyms “analysed” in collection in Europe D29: October 2010- Genotypes for true-to-type cultivars (= defined allelic pattern of varieties) D30: October 2010- List of identified local old, autochthonous cultivars maintained in collections in Europe M33 : December 2010 Analysis of significant citations reported in literature and on database about questionable or old autochthonous cultivars WP 6 Database improvement and enrichment Global objective of WP5 : Establishment of a sustainable EU-Viti-database, comprising all the data of Genres081 and the data recorded and elaborated within GrapeGen06 Objective 1 : Transformation of data already stored in the database according to new standards. Objective 2 : Improvement of data management capacities of the database Objective 3 : Increase of data content of the database Task 1 : Updating of existing data in the database (upadate of passeport data using EURISCO system) and entering passeport data for the new partners from Eastern Europe Task 2 : Improvement of the database structures and utilities (sorting features, identification of synonyms from coded SSR data,…..) Task 3: Transfer of new data in database (SSr, primary, secondary data and pictures, information on on-farm sites, data for V. sylvestris sites and individuals including SSR) Description of work, milestones and deliverables per year Year 1 :  Replacement of the former format by the Multicrop Passport Data Descriptor Format of the EURISCO-database, Update of the Genres081 partners collection’s passport data, addition of further collection’s passport data Year 2  Registration of primary, secondary descriptor and SSR-marker data and pictures as well as on- farm and in-situ data in a shared format  Elaboration and utilisation of a microsatellite comparison program for grape variety identity assessment in the examined material and automated verification of synonyms or misnaming. M34: January 2008 Registration data (passport, on-farm víneyards and in-situ sites of V. sylvestris, SSR-marker data). Installation of the coding and identification program for SSRs D31: January 2008 Data from year 1 for all WP published via the EU-Vitis-database Year 3  Registration of primary, secondary descriptor and SSR-marker data and pictures as well as on- farm and in-situ data in a shared format.  Establishment of differential levels of access to the database (publics, contributors, etc.)  Management of all new data and their introduction in the database in a shared format M35: January 2009 Registration data (passport, on-farm víneyards and in-situ sites of V. sylvestris, SSR-marker data, primary and secondary data and pictures). Registration of true-to type SSR-marker data D32: January 2009 Data published via the EU-Vitis-database Year 4  Registration of primary, secondary descriptor and SSR-marker data and fotos as well as on-farm and in-situ data in a shared format. Management of all new data and their introduction in the database in a shared format M36 : January 2010 Registration of data (primary and secondary data and true-to type SSR- marker data ) D33:- January 2010 Data published via the EU-Vitis-database M37: Registration of data (primary and secondary data and true-to type SSR-marker data ) D34: December 2010 Publication of a table summarizing the entered data subdivided by workpackages via the EU-Vitis-database (http://www.genres.de/eccdb/vitis) D35: December 2010 Dissemination of the results via wine growers magazines D36: December 2010 Publication of the meeting reports D37: December 2010 Publication of all data via the EU-Vitis-database WP 7 Elaboration of a long term conservation strategy Global objective of WP7 : Elaboration of a long term conservation plan for EU genetic resources Objective 1 : Development of alternative methodologies for conservation (cryo, on farm, ex-situ initiatives for V. sylvestris …) Objective 2 : Introduction of endangered cultivars and V. sylvestris accession in the collections Objective 3 : Safety duplication of variety and clonal material in collections Objective 5: Dissemination of data Task 1 : Development of cryo-conservation techniques and collections Task 2 : Introduction of material in collection (including safety duplication) or on-farm Task 3: Development of in-situ conservation procedures Task 4: Establishment of a management plan for Vitis resources – 2008-2018.

Description of work, milestones and deliverables per year Year 2  Cryopreservation: Selection of sites for pollen and seed collection , Collection of pollen and seed on the selected sites, Control of survival properties for the different samples  On-farm conservation: identification and agreements with growers interested in cooperation, for preservation, Preparation of material for new vineyard  Introduction in collection : collection of wood for the introduction D38: January 2008 Protocols for cryopreservation of pollen and seeds Year 3  Cryopreservation o Selection of sites already identified for pollen and seed collection, Test for the cryopreservation of buds from different origins, Collection of pollen and seed on the selected sites, Control of survival properties for the different samples o Organisation of a training course for cryopreservation techniques  On-farm conservation: Preparation of material for new vineyard, plantation of first vineyards  Introduction in collection : collection of wood for the introduction M38: Jan 2009 Definition of the list of growers and vineyards interested in preservation D39:January 2009 Protocols for cryopreservation of buds Year 4  Cryopreservation: Selection of sites already identified for pollen and seed collection, Test for the cryopreservation of buds from different origins, Collection of pollen and seed on the selected sites, Control of survival properties for the different samples  On-farm conservation: Plantation of vineyards  Introduction in collection : collection of wood for the introduction  Harmonization and duplication plan : according to the results of all WP, planification for safety duplications between the different collections 39: Jan 2010 Collection of pollen and seeds for cryopreservation and shipment of list to WP6 D40: January 2010 1st list of Vitis sylvestris accessions transferred in cryoconservation M40: Jan 2010 Collection of wood for transfert in collection and shipment of list to WP6 D41: January 2010 1st list of autochthonous cultivars transferred in on-farm conservation D42: January 2010 1st list of autochthonous cultivars and V sylvestris accessions transferred M41: Dec 2010 Collection of pollen, seeds and buds for Cryopreservation,shipment of list to WP6 D43: December 2010 Final list of Vitis sylvestris accessions transfered in cryoconservation M42: Dec 2010 Collection of wood for transfert in collection and shipment of list to WP6 D44: December 2010 Final list of old autochthonous cultivars transferred in on-farm conservation D45: December 2010 Final list old autochthonous cultivars and Vitsi sylvestris accessions transferred in collection M43: Dec 2010 Definition of the plan for duplication of the endangered germplasm in Europe D46: December 2010 List of endangered material in the european collections