The goal of this study was to examine the genetic difference among thirty two tomato accessions (Solanum lycopersicum L.) grown in Sulaymaniyah using ISSR molecular markers technique. The results of ISSR markers revealed 65 amplified fragments, 50 of them were polymorphic from using 15 primers. Fifteen ISSR markers used to detect DNA polymorphism gave polymorphism percentage for each primer range between 50-100% with an average polymorphism percentage reaching 75.61%. ISSR6 and Sola 11 gave the highest polymorphism percentage was 100%, while 3F, Sola 5 and Sola 12 did not give any amplification. The mean of PIC (Polymorphic Information Content) value was 0.50 for ISSR markers. The similarity matrix was obtained by using Jaccard's coefficients, based on polymorphic bands and dendrogram constituted through UPGMA cluster analysis. The dendrogram revealed 4 main groups. Genetic similarity (GS) ranged from 0.261 to 0.941 within studied accessions. The highest similarity was 0.941 for the genotype pairs G7 and G16. However, the lowest similarity index was 0.261 among G15-G24 and G15-G26. Tomato accessions were determined by two populations according to STRUCTURE analysis. The ISSR marker system ensured useful information in describing genetic diversity among tomato accessions collected from different geographical around Sulaymaniyah province.