Anthoxanthum aristatum Boiss. is native to southern and western Europe and North Africa. In Central Europe this species is recognized as invasive. Its ability to colonize various habitats may result from a high level of polymorphic isozyme loci and high morphological variability. The aim of this study was to recognize the degree of genetic diversity of the studied invasive species at its non-native range limit using molecular markers. The examined plant material consisted of 70 samples collected from 7 populations originating from various habitats: arable fields, recently abandoned field and sandy grassland. Ninety-six the Directed Amplification of Minisatellite-region DNA loci and 82 Sequence-Related Amplified Polymorphism loci were detected. The genetic diversity at the species level was high. The majority of genetic variation was present within populations (68% and 66%), while the remaining molecular variation was distributed among populations within and among habitat types. Specimens originating from sandy grassland populations were grouped together in one cluster that was genetically distinct from the arable field and fallow populations. The high genetic diversity populations at the A. aristatum non-native range limit in Central Europe and their marked diversity in arable field and sandy grassland habitats suggest that this species may systematically cross both geographical and ecological barriers.