Development of SSR (Simple sequence repeat) DNA markers in saffron (Crocus sativus L.) and their transferability to the closer relative species of saffron.

The aims of this study were to develop microsatellite markers for Crocus sativus (saffron) and to test them for the transferability into the close wild relatives of saffron. Four genomic libraries, which were enriched with different microsatellite motifs, were developed by using genomic saffron DNA of Safranbolu genotype. 350 of the 528 clones sequenced (% 66,3) contained microsatellites and 235 (%67,1) of the primers were designed. Optimum annealing temperature was determined for 188 (% 80.0) SSR markers with Gradient PCR protocol. The 21 (%16,4) out of 128 microsatellite primer pairs generated polymorphic bands among 27 saffron genotypes and 78 fragments were obtained. 52 of them were polymorphic. The average PIC value was 0.211, ranging from 0.071 to 0.499. Out of 138 SSRs developed, 114 in C.asumaniae, 129 in C.cartwrightianus, 121 in C. pallasi x C. mathewi, 124 in C. mathewi,122 in C. pallasii ssp. pallasii,122 in C. pallasii ssp. turcicus and 120 in C. pallasii ssp. dispathaceus gave amplicons. The results of phylogenetic analysis in taxa of Crocus series showed that the closest relative wild species of saffron could be C. cartwrightianus. From the results of the study, it was concluded that themicrosatellite markers developed in this study could bee useful both for evaluating genetic diversity in saffron and phylogenetic, taxonomic and evolutionary studies in Crocus species.