Evaluating Genetic Diversity of Iranian Alfalfa Local Populations Using Expressed Sequence Tags (ESTs) Microsatellites

To determine genetic diversity among some Iranian local varieties of alfalfa, six geographically diverse populations including: Bami, Rahnani, Nikshahri, Yazdi, Hamadani (from Isfahan), Hamadani (from Shiraz) along with Ranger, an American commercial variety, were evaluated using a set of 24 EST-SSR primers developed from cDNA library of Medicago truncatula and three microsatellite loci, identified from genomic library of M. sativa. Of the pairs of primers tested, four loci from EST-SSRs (AW9, BEE, TC6 and TC7) and genomic microsatellite (Afctt32), were found appropriate for assessing genetic diversity between these alfalfa genotypes. In total, 46 alleles were detected from the five loci in the samples of alfalfa examined. The number of alleles per locus in populations ranged from six to eleven and genetic diversity indices of loci were variable from 0.62 to 0.87 for the populations. Genetic relationship analysis of EST-SSR data revealed separation of Iranian populations from Ranger. It is likely that the parental origin of primary population from which Ranger has been derived is different from that of Iranian populations. Iranian local populations of alfalfa in this study were grouped in two main clusters. Alfalfa populations Hamadani and Rahnani, which are adapted to cold claimates, were grouped in one cluster and populations Bami, Yazdi and Nikshahri, belonging to the trpoical areas, were placed in the next cluster. The positioning of EST-SSR loci in coding regions of genome, possibly increases the usefulness of these markers to clarify inter specific genetic relationships among alfalfa populations.