Antarctic pearlwort ( Colobanthus quitensis ) is one of the flowering plant species considered native to maritime Antarctica. Although the species was intensively analyzed towards its morphological, anatomical and physiological adaptation to local environment, its genetic variability is still poorly studied. In the presented study, a recently developed retrotransposon−based DNA marker system (inter Primer Binding Site – iPBS) was applied to assess the genetic diversity and differentiation of C. quitensis populations from King George Island (South Shetland Islands, West Antarctic). A total of 143 scoreable bands were detected using 7 iPBS primers among 122 plant specimens representing 8 populations. 55 (38.5%) bands were found polymorphic, with an average of 14.3% polymorphic fragments per primer. Nine of all observed fragments were represented as a private bands deployed unevenly among populations. Low genetic diversity (on average H e = 0.040 and I = 0.061) and moderate population differentiation (F ST = 0.164) characterize the analyzed material. Clustering based on PCoA revealed, that the populations located on the edges of the study area diverge from the central populations. The pattern of population differentiation corresponds well with their geographic location and the characteristics of the sampling sites. Due to the character of iPBS markers, the observed genetic variability of populations may be explained by the genome rearrangements caused by mobilization of mobile genetic elements in the response to various stress factors. Additionally, this study demonstrates the usefulness of iPBS markers for genetic diversity studies in wild species.