The genus Pistacia is a dioecious and wind-pollinated member of the Anacardiaceae family that consists of 11 species. Among these species, Pistacia vera has edible nuts and commercial importance. The other species usually grow wild and are used as rootstock for .vera . Principally, the most typical wild Pistacia species in Iran are P.atlantica , P.khinjuk and P.vera L. var sarakhs. It is suggested that P.vera is the most ancestral species of Pistacia and the other species are probably its derivatives. Pistachio ( P.vera L.) cultivation and production, has a long history in Iran , and the country is well known internationally for this enterprise. Production and exportation of this product, in spite of the presence of international competitors, is comparatively beneficial and plays an important role in attracting foreign currency and attainment of economic trading status. Natural forests of wild Pistachio trees are still seen in different areas in North East of Iran and Afghanistan . They are valuable genetic resources and their genes can broaden the Pistachio gene pool and make the breeding programs more efficient. Moreover, Pitachio ability to tolerate salinity and drought conditions has made it a suitable crop for many regions of Iran . The multiple names for commercially grown Pistacia cultivars, utilized in different cultivation regions of Iran have somewhat created confusion for precise genotyping. Patterns of genetic diversity in Pistacia have been studied using a variety of morphological, physiological, and biochemical methods. Morphological characteristics vary among species. However, for many of the species, these distinctions are more difficult to evaluate. There is almost no cytogenetic information available on the various Pistacia species. Different researchers have conducted isozyme studies with the aim of distinguishing varieties of P.vera , but insufficient isozyme polymorphism among closely related cultivars has limited the usefulness of isozymes for analysis of genetic diversity and relatedness. The problem of insufficient polymorphism can be mitigated by using DNA-based markers. In this study, we attempted to 1) Demonstrate the characteristics of the trinucleotide (AAG) 8 microsatellites in order