GENETIC RESOURCES OF SAFFRON AND ALLIES (CROCUS SPP) The CROCUSBANK Project

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AGRI GEN RES 018 GENETIC RESOURCES OF SAFFRON AND ALLIES (CROCUS SPP) The CROCUSBANK Project José Antonio FERNANDEZ 1 What is Saffron? 2

3 The Genus Crocus include other species economically important 85 species Europe, North Africa and temperate Asia 4

The origin of saffron is Greece frescoes of Thera, Santorini 5 World production 6

Socio-economic aspects Highest priced spice 11,000 /kg (average 1,500-2,200 ) 55,000 Ha worldwide, 200-250 tones/year Saffron is profitable and competitive (non-subsidiary crop) Social importance Small-scale crop Stop declining populations in rural areas New uses related to human well-being 7 The quality and prestige of the market correspond to the European brands European Saffron under Protected Denominations of Origin (PDO): "Krokos Kozanis, Greece Azafrán de la Mancha, Spain "Zafferano dell'aquila", Italy Zafferano de San Gimignano, Italy Zafferano di Sardegna, Italy Zafferano delle Colline Fiorentine, Italy Munder Safran, Switzerland 8

Others pending: Safran du Gâtinais, France Safran du Quercy, France Safran de Provence, France Safran du Tarn et du Laugarais, France Safran de la Font Saint Blaise, France Zaferano di Cascia, Italy Azafrán del Jiloca, Spain Azafranes del Campo de Bello, Spain 9 Biodiversity of saffron is in danger Genetic erosion The CROCUSBANK Project 10

OBJECTIVES 1. Collection, multiplication, conservation and documentation of Crocus genetic resources i. Exploration and collection of germplasm of saffron and related species. ii. Elaboration of a list of descriptors for the characterisation of the genus Crocus. iii. Multiplication of the collected plant material for its conservation in the Bank of Plant Germplasm of Cuenca (Spain). iv. Elaboration of an effective documentation system. v. To make available this material to potential users by distribution of corms, tissue culture and DNA samples. 11 OBJECTIVES 2. Characterization and evaluation of Crocus genetic resources i. Morphological: Floral features, corm size ii.phenological: Flowering and relationship of climate, latitude and altitude iii.cytological characterization: Chromosome numbers, genome size, ploidy level and identification of hybrids iv.phytochemical: Chemical composition of saffron using chromatographic [GC/MS, HPLC/MS] and spectroscopic (IR, Raman, UV-Vis etc] techniques; metabolic profiling v. Molecular: AFLPs, SNPs, SSRs, etc. vi.abiotic stresses and pathogen responses 12

OBJECTIVES 3. Application of the Crocus germplasm information and banked accessions i. Rationalization of collections, identifying duplicates in order to optimise the management of the bank ii. Definition of valuable germplasm for saffron breeding iii. Identification of ecologically rare and important species/genotypes in the natural environment iv. Identification of valuable species, cultivars and hybrids for the horticultural industry v. Comparative genomics with model and crop species to identify universal features and valuable genes for agronomy 13 THE CONSORTIUM # 0: University of Castilla-La Mancha (UCLM, Spain) # 1: Junta de Comunidades de Castilla-La Mancha (JCCM, Spain) # 2: Agricultural University of Athens (AUA, Greece) # 3: Aristotle University of Thessaloniki (AUTH, Greece) # 4: Polytechnic University of Valencia (UPVLC, Spain) # 5: Tradimpex JM Thiercelin (TJMT, France) # 6: University of Catania (DOFATA-UNICT, Italy) # 7: University of Debrecen (UD; Hungary) # 8: National Polytechnic Institute of Toulouse (INPT, France) # 9: University of Leicester (ULEIC, UK) # 10: National Agricultural Research Foundation (Greece) (NAGREF, Greece) # 11: University of Kastamonu (formerly part of Gazi U.) (KU, Turkey) # 12: Azerbaijan National Academy of Sciences (ANAS, Azerbaijan) # 13: National Research Institute (Egypt) (NRC, Egypt) 14

WP01. Saffron Collection To collect saffron corms in geographical zones of saffron commercial cultivation (EU and abroad) and in zones of remaining minimal or relictic productions Expected: 160 accessions Obtained: 220 accessions 15 16

WP02. Crocus spp. Collection To collect seeds and/or corms of Crocus taxa from public and private collections, nurseries, botanic gardens and wild populations in the geographical areas of distribution of the genus Expected: 200 accessions Obtained: 352 accesions 17 18

WP03. Elaboration of a list of descriptors To elaborate a list of descriptors for the characterisation of the genus Crocus (including saffron) and morphological, phenological and physio-agronomical characterisation of the collected material 19 WP03. Elaboration of a list of descriptors Expected: 20 morphological descriptors Obtained: 111 20

WP04. Multiplication, conservation and documentation -The multiplication of the collected plant material for its conservation and management in the Bank of Plant Germplasm of Cuenca (Spain) -To guaranty an appropriate management of the Crocus germplasm collection and to make available this material to potential users 21 The Germplasm Bank of Cuenca, Spain 22

WP04: Results On farm conservation of the collections Reserve Vegetative Collection Exchange Vegetative Collection Rest Vegetative Collection Seed Collection 572 accessions covering 47 species are being preserved Documentation system working 23 24

WP05. Characterisation and evaluation -To characterise and evaluate genetic resources of C. sativus and allies take into consideration phenotypic characters with good heredability at different structural and physiological levels 62 C. sativusplus 68 non-saffron accessions have been evaluated at morphological, phenological and agronomical level 54 accessions were tested for susceptibility to salt stress Large genetic variability in saffron has been detected 25 Some details of morphological characterisation 26

Phytochemical characterisation 38 Saffron accessions and 31 other crocuses have been studied: FR-IR analysis Raman analysis Non-polar extracts from styles Volatile compounds GS-MS Response Surface Methodology (RSM) Colouring strength, bitterness & safranal content HPLC-DAD apocarotenoid analysis HPLC-DAD analysis of other metabolites Data under analysis 27 Molecular and Cytological characterisation 1. Cytological analysis of somatic metaphase chromosomes (different crocus spp.) 2. Cytological analysis of meiosis 3. AFLP markers (high variation) 4. SNPs markers (high variation) 5. IRAP & EST-ISSRs markers (no variation) Data under analysis 28

WP06. Application of the Crocus germplasm information and banked accessions To initiate exploitation of accessions of Crocus and morphological, agronomic and marker data -Identification of accessions of Saffron within the collection that are genetically identical -Quantification of diversity in Saffron and Crocus -Genomics in Saffron. The origin of saffron -Identification of germplasm for exploitation by breeders and growers 29 WP07. Project Management and Coordination -To co-ordinate and administrate the project -To monitor project s progresses -To evaluate project s results -To foster exploitation and dissemination of the results 30

The Consortium 31 In memoriam Fikrat Abdullaev 1943-2006 José Luís Guardiola 1943-2009 32

Thank you for your attention 33