Pistacia atlantica is one of the most widely distributed wild species in Iran. In the current study, the genetic diversity and genetic relationships among 15 natural populations of P. atlantica subsp. kurdica from northern Zagros forests in Iran were evaluated using simple sequence repeat (SSR) molecular markers. All the 14 SSR loci were polymorphic in the panel of 181 accessions. A total of 73 alleles were detected among the accessions with an average of 5.21 alleles per locus. The number of detected alleles (N) at each locus ranged from 3 to 7. The PIC values ranged from 0.58 to 0.81 with an average of 0.69. All the loci had PIC values higher than 0.50 which could be classified as very informative markers. Within the populations, Qhalaje and Hewraman populations had the highest and the lowest genetic diversity, respectively. The results of the overall AMOVA (P < 0.01) indicated that 64.00% of the variation was due to differences within populations and 36.00% occurred among populations. Genetic similarity showed a wide range of diversity (0.11–0.96), indicating high genetic variation among the accessions that originated from different populations. The dendrograms obtained from the SSR markers revealed high genetic variation among the studied accessions and grouped them into three main groups. Producers and researchers can use the provided data to enhance the preservation and reasonable utilization of the wild P. atlantica subsp. kurdica.