Tiller leaf anatomical features of nine Alopecurus species collected from high mountainous areas in Turkey were evaluated in the present study. Detailed leaf anatomical descriptions of the studied taxa and an identification key generated using the anatomical features are given. Clustering and principal coordinate analysis (PCA) methods were applied based on a total of 14 leaf anatomical characters. Anatomical differences between species were detected and anatomical characters of taxonomic interest were identified in this genus. With clustering analyses, a UPGMA tree was obtained to show the relationship between the species studied. Based on the results of PCA, the arrangement of adaxial sclerenchyma cells, the number of adaxial ribs, the number of abaxial sclerenchyma strands, the size of epidermal cells and the number of vascular bundles are designated as the most reliable characters to separate the species.