Development of SSR Markers in Lentil

The lack of informative simple sequence repeat (SSRs) markers in lentil (Lens culinaris M.) has been a major limitation in the lentil molecular breeding. Therefore, in order to develop SSR markers in lentil, four enriched genomic libraries for CA, GA, AAC and ATG repeats were constructed from the Lens culinaris cv Karacadağ. A total of 432 clone sequences were inquired for the detection of SSRs. The sequencing of 322 clones involved 360 repeat regions appropriated for designing primer pairs. A total of 360 SSR primers were designed and 149 of these were validated using eight Turkish lentil cultivars and seven Turkish landraces. 78 SSR primers produced total 400 polymorphic alleles, whereas 71 SSR primers amplified single expected sized allele. For 78 polymorphic SSR primers; the average number of alleles per locus was 5.1 and the polymorphism information content (PIC) value ranged from 0.06 to 0.89 with an average of 0.58. 51.2% and 24.3% of the polymorphic SSRs amplified the dinucleotide repeats and trinucleotide repeats, respectively. The most polymorphic repeat motifs were GA, CT, CA with a mean number of alleles per locus of 7.28, 7.14 and 5.83 respectively. Dinucleotide motifs such as GA, CT, TG could be the most useful appropriate target motifs to generate further SSR markers for lentil. Linkage map was constructed using 90 F₂ population derived from cross between Karcadağ x Silvan with 47 SSR markers. 43 out of 47 loci were assigned to six linkage groups. 40 out of 43 mapped microsattelite loci showed 1:1 expected Mendelian segregation pattern. The SSR markers developed in this study, could be for germplasm classification and identification, construction of genetic linkage maps, quantitative trait loci (QTL) mapping, association mapping, and marker assisted selection (MAS) in lentil