Background and Purpose: Genetic diversity is the material basis for crop improvement. In this study, genetic diversity of 30wheat genotypeswas evaluated at theDNAlevel using 24 simple sequence repeat (SSRs) markers. Materials and Methods: DNA extraction was performed according to the modified CTAB-method. Microsatellite analysis was performed using fluorescent fragment detection on a LI-COR 4200 DNA. Results and Discussion: The number of alleles per locus ranged from 1 to 14 with an average number of 8.44 alleles per locus. The highest number of alleles per locus was detected in the genome A with 7.2, compared to 5.9 and 5.0 for genomes B and D, respectively. The highest number of alleles was recorded at chromosome 7 (9.5), while the lowest number of alleles was detected at chromosomes 3 and 4 (5.0 and 5.3). The smallest genetic distance characterized genotypes Super Zitarka and Zitarka, Tena and Osjecanka, Tena and Bezostaja, Lela and Toras, Janica and Alka, Felix and Seka.Genotypes Pipi and Courtot showed the least genetic similiarities with rest of the genotypes. Conclusions: The identification of genetic diversity should be a good tool of selecting genotypes in breeding programs
