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¹ Batı Akdeniz Tarımsal Araştırma Enstitüsü, Antalya DOI : 10.16882/derim.2018.406434 Viewed : 2093 - Downloaded : 1014 This study was carried out to determine molecular characterization of annual button medic (Medicago orbicularis L.), which is very important for the pasture areas, collected from natural flora of Antalya province. Totally, the genetic diversity of 45 button medic genotypes were characterized with microsatellite (SSR) markers. A total of 35 microsatellite primer pairs were used in the preliminary study. In the second study, 15 pairs of microsatellite primers were used. Genetic similarity coefficients of the genotypes changed between 0.75 and 1.00. The closest genetic similarities were determined between genotypes 3-4-5 and 42; genotypes 37 and 38; 32 and 31 genotypes; genotypes 25 and 26; genotypes 7-9-16-18-22 and 33; genotypes 40 and 11; 29 and 30 genotypes; 13-14 genotypes and 6-10-12-15-17-19-20-21-23-24-28-3443-44 and 45 genotypes. The farthest similarity was determined between genotypes 2 and 25 and 26 genotypes. According to the UPGMA method, the genotypes in the dendogram were divided into two main groups at the level of similarity of 0.87. There were 12 genotypes in the first main group and 33 genotypes in the second main group. Keywords : Button medic; Antalya, Natural flora; Molecular characterization