A collection of 85 local Spanish peach cultivars has been studied with SSR markers. To carry out this work 42 SSRs previously developed in peach were screened to select the microsatellite loci that presented high polymorphism and that were easier to score in high resolution agarose gels. Ten SSRs were selected to identify the peach cultivars and to estimate the genetic similarity among them. The selected microsatellites resulted in the amplification of 35 fragments in the 85 genotypes studied, differentiating 46 genotypic profiles, from which 31 correspond to unique cultivars. The rest of the profiles correspond to groups of two to five cultivars and one group of 17 undistinguishable cultivars. The genetic similarity estimated from the molecular data clearly separated the flat and most of the white-fleshed peaches from the rest of genotypes that are mostly yellow non-melting local peaches. The results obtained in this work will help to improve the conservation and management of local Spanish peach germplasm allowing the establishment of core collections that represent most of the diversity conserved.