Thousands of landrace cultivars in wheat, rye, and triticale are stored in seed banks worldwide but the majority are inadequately described for their efficient exploitation in plant breeding. Studies on variability in these cereals using molecular markers have been relatively few and most of them were limited to regional genetic materials. In this study, a set of ISSR primers was used with samples of wheat, rye, and triticale accessions from various regions in the world to determine the level of genetic variation and relatedness among wheat, rye, and triticale accessions from the US National grain collection. The ISSR analysis revealed moderate level of polymorphic loci in wheat (40 %) and rye (41 %) accessions and high polymorphism (77 %) in triticale accessions. Nei's gene diversity (h) was 1.4, 1.4, and 1.77, for wheat, rye, and triticale, respectively. The same trend was observed for Shannon's index (I), the effective (Ne) and observed number (Na) of alleles with the triticale showing the highest values. The genetic distance data showed that the wheat, rye and triticale accessions from different provenances were genetically close. Overall, the constructed dendrograms revealed no clustering related to geographic provenance of the accessions analyzed. Some accessions from different regions showed similar amplification profiles.