The Drake Passage arises as a likely route for gene flow into Antarctica, as it is the shortest path between this continent and the rest of the world. Despite this, long-distance dispersion into Antarctica could be particularly complex for terrestrial biota. To compare the levels of genetic diversity between Antarctic and South American populations of the Antarctic pearlwort, Colobanthus quitensis, we conducted the first estimation of genetic diversity in this species using amplified fragment length polymorphism. Four populations across the Drake Passage were selected and their genetic composition was characterized. Differences among the levels of genetic diversity were found between the populations analysed as well as between their allelic identities. However, interestingly, their spatial distribution across the Drake Passage suggests a north-to-south gradient of increasing genetic diversity.