Genus Allium L. contains very taxonomically complex sections, especially the subgenus Melanocrommyum. The systematic position of the species in each section has been revised many times over time. In the present study, the relationship between 32 ecotypes belonging to 10 different species of Allium was investigated using ISSR markers. The nine primers used produced 166 polymorphic bands (average 18 bands per primer). Among the primers used, ISSR873 primer with 27 bands made the most, and ISSR4 primer with two bands had the lowest polymorphic bands. The PIC of the markers ranged from 0.04 to 0.43. Cluster analysis by UPGMA method based on molecular markers divided the studied ecotypes into four groups. The clustering and principal coordinate analysis results showed that most morphologically similar species were grouped in closed clusters. According to Dice similarity coefficient, the highest percentage of similarity was shown between Allium stipitatum and Allium saralicum ecotypes (72 percent) from the Melanocrommyum subgenus, and the lowest similarity was obtained between Allium tripedale and Allium iranicum ecotypes (12 percent). The ecotypes with the lowest similarity percentage belong to the subgenus Allium and Nectaroscordum, which are placed in separate clusters. Based on the results, the ecotypes of Pseudoprason, Melanocrommyum, and Procerallium sections showed the highest affinity. In general, it can be concluded that ISSR markers are useful for classifying Allium species and have sufficient potential for phylogenetic studies of species. In addition, due to significant genetic diversity among the studied ecotypes of wild Allium species, this diversity can be used in future breeding programs of crop