Systematics of the Widely Distributed Japanese Clouded Salamander, Hynobius nebulosus (Amphibia: Caudata: Hynobiidae), and Its Closest Relatives

Abstract The abundant clouded salamander from western Japan, Hynobius nebulosus, has long been considered a single widespread species, although some authors have suggested the inclusion of several cryptic species. This led to a molecular and morphological analysis of populations from all parts of the known range. Phylogenetic relationships were inferred from complete sequences of the mitochondrial cytochrome b gene, and nuclear genome differentiations were estimated by multiplexed inter simple sequence repeat genotyping by sequencing (MIG-seq). The results suggest that H. nebulosus hitherto recognized consists of at least nine species. We applied existing names to two of them, H. nebulosus (Temminck et Schlegel) and H. vandenburghi Dunn, and described seven others as new species.