The topology also agreed with that of the equally parsimonious 100 trees (length: www.selleckchem.com/products/BAY-73-4506.html 269; CI: 0.903; RI: 0.990; RC: 0.894) obtained by the maximum parsimony method. In these trees, three groups, corresponding to groups A, B, and C in Figure 2, diverged simultaneously at the basal node. Subgroups at the lower level were recognized only in the populations that originated in the Sakishima region. All these groupings were supported strongly by the bootstrap analyses (99-100%).Figure 3Neighbor-joining tree based on the Jukes-Cantor distances calculated using the 856bp long data set of the nuclear rDNA ITS2 sequence. Different sequences obtained from the same individuals were distinguished by lower-case letters, a and b, included …4.

DiscussionIn this study, we found that Japanese Curtos fireflies were divided among three genetically separated local populations, corresponding to C. costipennis in the Amami region, C. okinawanus in the Okinawa region, and C. costipennis in the Sakishima region (Figures (Figures22 and and3).3). Although the nucleotide sequences of C. okinawanus formed a monophyletic group (group B), those of C. costipennis were separated between two distinct groups, A and C, and the monophyly of this species was not recognized in analyses using both mtDNA and nuclear DNA sequences (Figures (Figures22 and and3).3). In addition, the mtDNA haplotypes of C. okinawanus (group B) were positioned between those of C. costipennis (groups A and C) (Figure 2). These results indicate that the currently recognized C. costipennis species is paraphyletic. Otherwise, C.

okinawanus must be a subspecies or a geographic strain included within the single species C. costipennis.Although the two species closely resemble each other in both morphology and behavior [7], they are apparently distinct in terms of elytral coloration (C. costipennis: dark yellow except for elytral apex, C. okinawanus: entirely brownish black), and this characteristic is recognized as diagnostically important [6]. However, the present study revealed that such a characteristic did not reflect evolutionary relationships correctly. On the other hand, several researchers recognized variations in body size, the shape of the male genital organ, and the number of punctures on the elytra among local populations of C. costipennis [5, 6]. Reevaluation of these characteristics is needed to confirm the taxonomic status of the two species.Among the three major groups detected in this study, a closer relationship was recognized between C. costipennis and C. okinawanus distributed in neighboring regions (the Amami and Okinawa regions in the Central Ryukyu Islands) Brefeldin_A than between the populations of C.