| Summary: | Here, we describe a new species of the genus <i>Hynobius</i> from Chugoku, Japan. In populations from central to eastern Shimane Prefecture, the Izumo Lineage of <i>Hynobius utsunomiyaorum</i> was clearly distinguished from the true <i>Hynobius utsunomiyaorum</i> based on morphological and molecular evidence. Thus, we describe the former lineage as a new species, <i>Hynobius kunibiki</i> sp. nov. Morphological comparisons revealed that <i>H. utsunomiyaorum</i> lacks a distinct yellow line on the ventral side of its tail, whereas the new species possesses this yellow line; most <i>H. utsunomiyaorum</i> individuals have distinct white spots on the lateral sides of their body and lack a fifth toe, whereas the new species largely lacks these spots, and all examined individuals had a fifth toe. The two species also differed significantly by several other morphological characteristics. The lentic species <i>Hynobius setoi</i> is morphologically similar to <i>H. kunibiki</i> sp. nov., but they differ significantly by various morphological characteristics. Despite their partial morphological similarity, these two species differed substantially in terms of their genetics. Finally, we show, in a phylogenetic tree including all Japanese <i>Hynobius</i> species, that the subgenus <i>Hynobius</i> can be divided into four genetic clades. Overall, this information will help develop conservation management strategies and policies for these species.
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