Vertebrates usually reproduce sexually in which males and females contribute their offspring genome and produce genetically diverse offspring. However, some of them are asexual without genetic contribution from males. The nocturnal gecko, Lepidodactylus lugubris, is all females and reproduces parthenogenetically. This gecko is known to consist of diploid and triploid clones in the tropical and subtropical regions, which can be identified by their dorsal marking patterns, ploidy, and protein polymorphism. This gecko is also distributed in the southern parts of Japan, and several clones have been reported. In this study, we investigated the origins and genetic diversity of Japanese L. lugubris by clonal discrimination using microsatellite and mitochondrial DNA analyses. A total of 748 individuals were collected from 21 islands of five island groups (Ogasawara, Okinawa, Miyako, Yaeyama and Daito Islands) and 17 clones were distinguished genetically. Mitochondrial cyt b sequences of these clones suggested that they were all closely related and differentiated recently. Clonal diversity was much higher (14 clones) in the Daito Islands than in the other island groups in which only one or two clones coexisted. Judging from the dorsal marking patterns and ploidy known so far, six clones were cosmopolitan and may be colonized from the outside of Japan. However, other 11 clones were endemic to the Daito Islands and explained by possible hybridization between the one female diploid clone and one male diploid clone because other 9 clones were triploid and all had the combinations of polymorphic microsatellite alleles of these female and male diploid clones. Although the males have never been recorded in the Daito Islands, males might appear in the past. These findings contribute to understanding of clonal diversity and dynamics of asexually reproducing animals. If diploid parthenogenetic geckos can produce triploid clones by mating with the diploid males, clonal diversity would increase rapidly in a small island, and such newly produced triploid clones would expand widely. K E Y W O R D S asexual reproduction, clonal diversity, hybridization, microsatellite analysis, ploidy change