Natsuhiko Yoshikawa

Phylogenetic relationships and biogeography of the Japanese clawed salamander, Onychodactylus japonicus (Amphibia: Caudata: Hynobiidae), and its congener inferred from the mitochondrial cytochrome b gene

Using the mitochondrial cytochrome b gene, we investigated phylogenetic relationships between and within the Japanese clawed salamander, Onychodactylus japonicus, and its close continental relative O. fischeri. Monophyly of O. japonicus was well supported, and O. japonicus was clearly distinguished from O. fischeri. However, O. fischeri comprises genetically distinct populations from Russia, NE China, and Korea that do not form a monophyletic group. Within O. japonicus, four major clades were clearly recognized: Clade I from northern Tohoku district, Clade II from southern Tohoku district and the Tsukuba Mountains, Clade III from southwestern Honshu, and Clade IV from Kinki and Chugoku districts in Honshu and from Shikoku. Although genetic distances among these clades were large (5.5-9.6%), relationships among the clades were unresolved. All clades except Clade I contained two or three distinct subclades. In several localities in Kinki and Chugoku, Clades III and IV were sympatric. The estimated divergence times and available geohistorical data suggest that O. japonicus began to differentiate in the Upper Late Miocene and that the pattern of genetic differentiation of this species has been affected strongly by climate changes and geohistorical events such as volcanic activity and mountain formation. Our results suggest that both O. fischeri and O. japonicus comprise multiple cryptic species.

Phylogenetic relationships of two Salamandrella species as revealed by mitochondrial DNA and allozyme variation (Amphibia: Caudata: Hynobiidae)

We conducted molecular phylogenetic analyses to confirm taxonomic relationships and to delimit distributional ranges of Siberian salamanders, Salamandrella keyserlingii and Salamandrella schrenckii, and to elucidate the origin of the isolated population of this species complex on Hokkaido, Japan. Phylogenetic trees constructed by MP, NJ, ML, and Bayesian methods, using complete sequences of mitochondrial cytochrome b genes, all indicated monophyly of Salamandrella and of each of the two species. Identical relationships were found on UPGMA, NJ, and CONTML trees derived from electrophoretic analysis of variation in 18 inferred allozyme loci. Populations from Hokkaido and northeastern China proved to be S. keyserlingii, while populations from Khabarovsk and Lazovsky are S. schrenckii. Genetic differentiations of S. keyserlingii within Sakhalin, and between Sakhalin and Hokkaido, are substantial. The Hokkaido population is hypothesized to have been isolated on the island since early Pleistocene, much earlier than isolation of sympatric anuran populations from their Sakhalin relatives. In contrast, the continental populations of S. keyserlingii are only slightly differentiated from some Sakhalin populations, and are thought to have expanded their ranges in the late Pleistocene.

Molecular phylogeny and biogeography of caecilians from Southeast Asia (Amphibia, Gymnophiona, Ichthyophiidae), with special reference to high cryptic species diversity in Sundaland

We investigated the phylogenetic relationships and estimated the history of species diversification and character evolution in two ichthyophiid genera: Caudacaecilia and Ichthyophis. We estimated the phylogenetic relationships of 67 samples from 33 localities in Southeast Asia from 3840-bp sequences of the mitochondrial 12S rRNA, 16S rRNA, and cyt b genes using Bayesian inference, maximum likelihood, and maximum parsimony methods. The Southeast Asian samples formed a well-supported clade differentiated from a South Asian sample. The Southeast Asian clade was divided into two subclades, one containing samples from South China, Indochina, Malay Peninsula, and Java. The other consisted of samples from Borneo and the Philippines. Neither Caudacaecilia nor Ichthyophis was monophyletic, nor did samples with or without light stripes lateral to the body form clades. We found several distinct sympatric lineages and undescribed species, especially from Sundaland.

Phylogeny and historical demography of Cynops pyrrhogaster (Amphibia: Urodela): taxonomic relationships and distributional changes associated with climatic oscillations.

We investigated the phylogenetic relationships and estimated the historical demography of the Japanese fire-bellied newt, Cynops pyrrhogaster, from Japanese mainlands using 1407-bp sequences of the mitochondrial DNA (NADH6, tRNAglu, cyt b) and 1208-bp sequences of nuclear DNA (Rag-1) genes. Phylogenetic trees based on mitochondrial DNA revealed four major haplotype clades (NORTHERN, CENTRAL, WESTERN, and SOUTHERN clades) within this species. Degree of genetic differentiation among major haplotype clades was very large for intraspecific variation, suggesting this species to be composed of four species lineages that replace each other geographically. Nuclear genetic variation presented no obvious patterns of geographic structure except for the distinctness of populations diagnosed by NORTHERN clade of mitochondrial haplotypes, suggesting results of incomplete lineage sorting. Current distribution and estimated divergence times for the genus Cynops suggest that the common ancestor of two Japanese species (C. pyrrhogaster and C. ensicauda from the Ryukyu Islands) had diverged at the edge of the continent corresponding to the present East China Sea and Central Ryukyus. Subsequent range expansion to Japanese mainland seems to have occurred in the middle Miocene. Population-genetic analyses indicated that all species lineages, except for the SOUTHERN one, experienced geographic population reductions and expansions associated with glacial and postglacial climatic oscillations.

Allozymic Variation in the Japanese Clawed Salamander, Onychodactylus japonicus (Amphibia: Caudata: Hynobiidae), with Special Reference to the Presence of Two Sympatric Genetic Types

We conducted an electrophoretic survey to examine geographic genetic variation in samples from 17 localities of the Japanese clawed salamander, Onychodactylus japonicus. This species was divided into six genetic groups (N-Tohoku, S-Tohoku, Tsukuba, SW-Honshu, Kinki, and Shikoku) that were largely concordant with clades or subclades recognized in our previous mtDNA study. Although the relationships among these six groups were not clarified, genetic distances between them were not small (mean Nels D=0. 146–0.471). Among these groups, the geographically isolated Tsukuba group was genetically distinct, possibly as a result of population isolation. In a locality of western Honshu, two groups, SW-Honshu and Shikoku, were found to occur sympatrically. Although several presumable hybrid individuals were found, hybridization between these two groups seemed to occur very rarely. These results indicate that the Shikoku group is specifically distinct from the SW-Honshu group, whose range includes the type locality of O. japonicus.

Allozymic Variation and Phylogeography of Two Genetic Types of Onychodactylus japonicus (Amphibia: Caudata: Hynobiidae) Sympatric in the Kinki District, Japan

On the basis of allozyme and mtDNA sequence variation, we elucidated genetic relationships between two sympatric genetic types of Onychodactylus japonicus in Kinki and adjacent districts, and investigated their phylogeography. Allozymic analysis revealed the presence of two distinct genetic types (the SW-Honshu and Kinki groups) in this area, and their sympatric occurrence in three of 10 sampling sites. Fixed or nearly fixed allele differences in several loci strongly suggested reproductive isolation between the two types, although one hybrid specimen was found in a locality. Analyses of mtDNA using 194 specimens from 22 localities also demonstrated two genetic types. From phylogeographic and population genetic analyses, it was surmised that these two types diverged allopatrically, and secondarily contacted to become sympatric by the Pleistocene uplift of mountains. Our results indicate different specific status for these two types and separation of the Kinki group from O. japonicus, to which the SW-Honshu group belongs.

First Record of the Tree-Frog Genus Liuixalus from Vietnam with the Description of a New Species (Amphibia: Rhacophoridae)

Abstract: We record a tree frog of the genus Liuixalus for the first time from outside of China and describe it as a new species, Liuixalus catbaensis, on the basis of a single juvenile specimen collected from Cat Ba Island, northern Vietnam. The new species is easily distinguished from all other members of the genus Liuixalus by its uniformly brick-red dorsum lacking dark markings. The biogeographical significance of finding of this species in Vietnam is briefly discussed.

A New Salamander of the Genus Onychodactylus from Tsukuba Mountains, Eastern Honshu, Japan (Amphibia, Caudata, Hynobiidae)

Abstract: Recent phylogenetic studies using mtDNA and allozymes have revealed the presence of large genetic differentiation within a Japanese clawed salamander, Onychodactylus japonicus, suggesting the presence of several cryptic taxa in this species. Based on morphological analyses, we describe one of them from the Tsukuba Mountains of Ibaraki Prefecture, in the Kanto district of eastern Honshu, as a new species, Onychodactylus tsukubaensis. It is a member of the japonicus species complex of Onychodactylus, and differs from the other species of the complex by a relatively short tail, wide head, and large number of vomerine teeth.

Two new Salamanders of the genus Onychodactylus from Eastern Honshu, Japan (Amphibia, Caudata, Hynobiidae)

We describe two new species of hynobiid salamanders in the genus Onychodactylus from eastern Honshu, Japan, based on the morphological and genetic evidence. Onychodactylus intermedius sp. nov. is distributed in southern part of Tohoku District and northern Ibaraki and Niigata Prefectures, and was previously reported as S-Tohoku group. Onychodactylus intermedius belongs to the O. japonicus species complex, and differs from the other congeners in having relatively long tail, narrow head, short snout, 18 presacral vertebrae, and distinctly curved vomerine tooth series without gap. Onychodactylus fuscus sp. nov. is known from only four localities in Fukushima and Niigata Prefectures of Tohoku and Hokuriku Districts. It also belongs to the O. japonicus complex, but lacks the dorsal stripe, which is a diagnostic character of the species complex. In other characteristics, O. fuscus differs from the other congeners in having comparatively long tail, wide head and internarial space, shallowly curved vomerine tooth series with gap, and relatively few vomerine teeth. Both species described here breed in winter. Phylogenetically, the two new species are closely related to each other, forming a well-supported clade with O. tsukubaensis as their sister species. Onychodactylus intermedius sp. nov. is known to be parapatric with O. japonicus and O. nipponoborealis without hybridization, whereas O. fuscus sp. nov. is sympatric with O. japonicus at least in a single known locality, and analysis of microsatellite loci indicates they are reproductively isolated.

Stock Origin of the Leatherback Turtles (Dermochelys coriacea) Found in the Vicinity of Japan Revealed by mtDNA Haplotypes

Abstract: We determined sequences of mitochondrial control region of 16 specimens of the leatherback turtles, Dermochelys coriacea, collected in the vicinity of Japan, and estimated stock origin of Japanese migrants. As a result, we found three haplotypes (JD1, 2, and 3) in these specimens, and 14 specimens possessed JD1. In comparison with previously reported sequences, JD1 and 2 were identical to a single haplotype endemic to West Pacific stock in New Guinea and Solomon Islands, whereas JD3 was identical to a haplotype nearly endemic to Malaysian stock. Our results indicate that most of the leatherbacks foraging around Japan originate in West Pacific stock, and a few may migrate from the Malaysian nesting population.