Mitochondrial Cytochrome b Phylogeny and Historical Biogeography of the Tohoku Salamander, Hynobius lichenatus (Amphibia, Caudata)

The Tohoku salamander, Hynobius lichenatus Boulenger, 1883, is a lentic breeding species widespread throughout montane regions of northeastern Japan. To explore intraspecific genetic variation and infer evolutionary history of H. lichenatus, we performed mitochondrial DNA analysis (complete 1141 bp sequences of the mitochondrial cytochrome b gene) using 215 adult and larval individuals collected from 75 localities, encompassing known distributional range of the species. Hynobius lichenatus proved to be monophyletic, including three well-supported and geographically structured clades (Clade I from northern Kanto, Clade II from southern Tohoku, and Clade III from northern Tohoku). These clades, respectively, comprise several subclades, and show genetic distances as large as those seen between different species of Hynobius. Results of population statistic analyses indicate that all clades and most subclades have maintained high genetic diversity and demographic stability over long periods. Molecular dating indicates divergence in H. lichenatus concords with topographic evolution of northeastern Japan from late Miocene to early Pleistocene, suggesting that paleogeographic events in this region, such as orogenesis, sea level change, and volcanic activity, have been crucial for shaping genetic patterns and diversity in this species. Hynobius lichenatus greatly differs from many other animal species from northeastern Japan in its much older periods and the pattern of genetic differentiation, and is suggested as an old faunal element in this region.