The genetic polymorphism of thirty Aegilops ventricosa Tausch. accessions sampled from northern Algeria was evaluated using inter-simple sequence repeat (ISSR) markers (into 5’). Six reproducible primers were used, and 53 out of 62 amplified distinct bands were found with a high score of polymorphism (87.48%). A significant number of specific bands was identified which were useful for genotyping. The majority of ISSR primers showed important resolving power with an average of 10.86, reflecting high informative markers tested in this investigation. SIMQUAL coefficient was used to calculate the genetic similarity among the 30 accessions. This similarity ranged from 0.5806 to 1. Based on Unweighted Pair-Group Method using Arithmetic Averages (UPGMA) method, a dendrogram was established to assess the intraspecific relationships via the genetic distances between the 30 accessions. The ISSR results showed a high variability in the studied collection. These markers could be efficiently applied to estimate the genetic diversity in the Ae. venticosa or others species in the Aegilpos genus.