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Climate change is one of the major threats to global amphibian diversity, and consequently, the species distribution is expected to shift considerably in the future. Therefore, predicting such shifts is important to guide conservation and management plans. Here, we used eight independent environmental variables and four representative concentration pathways (RCPs) to model the current and future habitat suitability of the Korean clawed salamander (Onychodactylus koreanus) and then defined the dispersal limits of the species using cost distance analysis. The current habitat suitability model generated using the maximum entropy algorithm was highly consistent with the known distribution of the species and had good predictive performance. Projections onto years 2050 and 2070 predicted a drastic decrease of habitat suitability across all RCPs, with up to 90.1% decrease of suitable area and 98.0% decrease of optimal area predicted from binary presence grids. The models also predicted a northeastward shift of habitat suitability toward high‐elevation areas and a persistence of suitability along the central ridge of the Baekdudaegan Range. This area is likely to become a climatic refugium for the species in the future, and it should be considered as an area of conservation priority. Therefore, we urge further ecological studies and population monitoring to be conducted across the range of O. koreanus. The vulnerability to rapid climate change is also shared by other congeneric species, and assessing the impacts of climate change on these other species is needed to better conserve this unique lineage of salamanders.

The raccoon dog (Nyctereutes procyonoides), endemic to East Asia, is classified as six subspecies according to their geographical distribution including a population introduced to Europe. Studies on phylogenetic relationship or population genetics in both native and introduced areas have been carried out recently. Lately, opinions that Japanese raccoon dogs should be classified as a different species were asserted based on several studies using karyotypes, morphometric characters, mtDNA, and microsatellites analysis. However, no data pertaining to the nuclear DNA (nDNA) or Y chromosome are available. To estimate the relationship among the species using different genes is necessary in understanding of the history of this species. Therefore, we investigated nDNA and Y chromosomes in our study to define relationships: (1) between continental raccoon dog populations, (2) between original and introduced groups, and (3) between continental and Japanese groups. The analysis of four nuclear (CHRNA1, VTN, TRSP, WT1) and ZFY genes indicated that there had been no genetic differentiation among the continental populations. However, significant differences were observed between continental and Japanese raccoon dogs in VTN and ZFY genes implying genetic differentiation has been going between them. To better understand the phylogenetic relationship among raccoon dog populations, further study will be necessary.

The Japanese otter (Lutra nippon), once inhabited in most islands of Japan, is now considered as an extinct species. Although the Japanese otter is regarded as a distinct species from the Eurasian otter (L. lutra), its phylogeny and taxonomic status are based on limited information on morphological and genetic data, and thus further clarification is required. Here, we assessed the phylogenetic relationship among the genus Lutra and taxonomic status of L. nippon by using the complete sequences of cytochrome b gene of its holotype. The present phylogenic trees supported that the genus Lutra specimens largely formed monophyletic group, with L. sumatrana as a basal to other Lutra species. Within Lutra species, L. nippon was distantly related with L. lutra. The European otter population of L. l. lutra were clustered together with its subspecies, L. l. chinensis rather than the same subspecies, Korean otter population. The discrepancy between the genetic data and traditional taxonomy justifies the necessity of reexamination of the current subspecific classification system of Eurasian otters. Level of genetic divergence between the holotype of L. nippon and L. lutra was two to three-fold lower than those among the other sister species of the Lutrinae. Based on the level of divergence between the L. nippon and L. lutra, and insufficient evidence of morphological difference between them, it is suggested that designation of Japanese otter as a separate species from L. lutra will be reconsidered.

Simple Summary Morphological changes are common in populations of animals in response to environmental and evolutionary forces. This is the case for salamanders, which can adapt to most environments on earth. On the Korean Peninsula, Hynobius salamanders are widespread, with several species overlapping in distribution. In addition, while there are seven segregated clades based on mitochondrial DNA, only four of them have been described as segregated species and the three others are candidate species for which the species level of divergence has not been tested yet. Here we measured 329 individuals from all seven clades, in areas of range overlap or not, on islands and on the mainland (Graphical Abstract A), and tested for the species status of the three candidate species. Individuals on the mainland had a generally broader morphology than those on the islands (Graphical Abstract B), and individuals in the range overlap differed from the individuals from the same species that were not found in presence of another clade (Graphical Abstract C). Despite a significant impact of the island effect and the sympatric areas, all seven clades have significantly different morphologies, and we describe Hynobius notialis sp. nov., Hynobius geojeensis sp. nov. and Hynobius perplicatus sp. nov. Abstract Closely related individuals from different areas can see their morphologies change based on differences between clades, but also ecological variables such as the island effect or sympatry. This is the case of salamanders, which have adapted to a broad range of ecological niches, ranging from underground dwellers in xeric landscape to tropical arboreal habitats. On the Korean Peninsula, salamanders from the Hynobius clade are widespread on the mainland and islands, with several partially sympatric clades and candidate species. Currently, seven lineages have been identified based on mtDNA, four of them matching named species and three others for which the species status remains untested. While the morphology of Korean Hynobius is known to be variable between genetically segregated clades, we hypothesise that (1) the candidate species are morphologically different, and that (2) the island effect and (3) the sympatric status have significant impacts on the morphology of individuals within the genus. Here we measured 329 Hynobius salamanders from all seven clades, in areas of sympatry and allopatry, and on islands and on the mainland (Graphical Abstract A). We determined that the island effect had a significant impact on the morphology of the genus, with mainland individuals generally displaying a broader range of morphology than islandic individuals (Graphical Abstract B). We also determined that sympatry had an impact on morphology, with the sizes of individuals from clades in sympatric areas diverging from each other (Graphical Abstract C). Finally, we demonstrated that all seven clades have significantly different morphologies, and we described the three candidate species that had already been isolated based on mtDNA and microsatellite data: Hynobius notialis sp. nov., Hynobius geojeensis sp. nov. and Hynobius perplicatus sp. nov. We conclude that looking at morphology alone would be misleading about the true diversity of Hynobius species, and species in general, because of the island and patry effects.

We conduct a phylogeographic and population genetic study of the Asiatic Toad (Bufo gargarizans) to understand its evolutionary history, and the influence of geology and climate of the region. A total of 292 individuals from 94 locations were genotyped for two mitochondrial DNA loci (cytb, ND2 gene) and five nuclear introns (Sox9-2, Rho-3, CCNB2-3, UCH-2, DBI-2). We performed a suite of phylogenetic, population genetic, and divergence dating analyses. The phylogenetic trees constructed using mitochondrial loci inferred B. gargarizans being divided into two major groups: West (China mainland) and Northeast (Northeast China, Russia and Korean Peninsula). As with previous studies of this species, we recover population genetic structure not tied to geographic region. Additionally, we discover a new genetic clade restricted to Northeast Asia that points towards the Korean Peninsula being a glacial refugium during the Pleistocene. The weak phylogeographic pattern of B. gargarizans is likely the result of multiple biological, anthropogenic, and historical—robust dispersal abilities as a consequence of physiological adaptations, human translocation, geologic activity, and glacial cycles of the Pleistocene. We highlight the complex geologic and climatic history of Northeast Asia and encourage further research to understand its impact on the biodiversity in the region.

While comparatively few amphibian species have been described on the North East Asian mainland in the last decades, several species have been the subject of taxonomical debates in relation to the Yellow sea. Here, we sampledDryophytessp. treefrogs from the Republic of Korea, the Democratic People's Republic of Korea and the People's Republic of China to clarify the status of this clade around the Yellow sea and determine the impact of sea level change on treefrogs' phylogenetic relationships. Based on genetics, call properties, adult morphology, tadpole morphology and niche modelling, we determined the segregated status species ofD.suweonensisandD.immaculatus. We then proceeded to describe a new treefrog species,D.flaviventrissp. nov., from the central lowlands of the Republic of Korea. The new species is geographically segregated fromD.suweonensisby the Chilgap mountain range and known to occur only in the area of Buyeo, Nonsan and Iksan in the Republic of Korea. While the Yellow sea is the principal element to the current isolation of the three clades, the paleorivers of the Yellow sea basin are likely to have been the major factor for the divergences within this clade. We recommend conducting rapid conservation assessments as these species are present on very narrow and declining ranges. Conservation Research grant from The Biodiversity Foundation; National Research Foundation of KoreaNational Research Foundation of Korea [2017R1A2B2003579]; National Institute of Biological Resources (NIBR) - Ministry of Environment (MOE) of the Republic of Korea [NIBR201803101] This work was supported by a Conservation Research grant from The Biodiversity Foundation to AB, a research grant from the National Research Foundation of Korea (2017R1A2B2003579) to YJ, and by a grant from the National Institute of Biological Resources (NIBR), funded by the Ministry of Environment (MOE) of the Republic of Korea (NIBR201803101) to MSM and TK.

The water toad (Bufo stejnegeri) is endemic to Northeastern Asia (South Korea, North Korea, and China) and has unique ecology for a toad by being semi-aquatic and breeding in lotic environments. We use a suite of phylogenetic analyses to understand the evolution of B. stejnegeri's distinctive ecology and the impact of Pleistocene glacial cycles on the biodiversity of Northeast Asia. For the evolution of a semi-aquatic lifestyle, although B. stejnegeri is relatively closely related to two other semi-aquatic Bufo species (B. torrenticola and B. andrewsi), ancestral state reconstruction analysis infers an independent evolution in all three species. Upon closer inspection, B. stejnegeri exhibits major differences in amplexus and egg-laying behavior compared to the other two species, supporting independent evolution. Divergence dating analyses infer B. stejnegeri to have originated during the Late Miocene-Early Pliocene (4.3 Ma, 2.7–6.2 Ma). This species does not exhibit population differentiation with respect to mountain range, but shows a preliminary genetic pattern of southern richness and northern purity supporting a single refugium in Korea during Pleistocene glacial cycles. The Bayesian skyline plot supports this inference, suggesting a population decline followed by expansion during the Pleistocene. Although not as species rich as the tropics, we hope this study helps spark interest in Northeast Asian biodiversity.

Many peninsulas in the temperate zone played an important role as refugia of various flora and fauna, and the southern Korean Peninsula also served as a refugium for many small mammals in East Asia during the Pleistocene. The Asian lesser white-toothed shrew, Crocidura shantungensis, is a widely distributed species in East Asia, and is an appropriate model organism for exploring the role of the Korean Peninsula as a refugium of small mammals. Here, we investigated phylogenetic relationships and genetic diversity based on the entire sequence of the mitochondrial cytochrome b gene (1140 bp). A Bayesian tree for 98 haplotypes detected in 228 C. shantungensis specimens from East Asia revealed the presence of three major groups with at least 5 subgroups. Most haplotypes were distributed according to their geographic proximity. Pairwise FST's and analysis of molecular variance (AMOVA) revealed a high degree of genetic differentiation and variance among regions as well as among populations within region, implying little gene flow among local populations. Genetic evidence from South Korean islands, Jeju-do Island of South Korea, and Taiwan leads us to reject the hypothesis of recent population expansion. We observed unique island-type genetic characteristics consistent with geographic isolation and resultant genetic drift. Phylogeographic inference, together with estimates of genetic differentiation and diversity, suggest that the southern most part the Korean Peninsula, including offshore islands, played an important role as a refugium for C. shantungensis during the Pleistocene. However, the presence of several refugia on the mainland of northeast Asia is also proposed.

The prevention and control of infectious diseases transmitted by wildlife are gaining importance. To establish effective management strategies, it is essential to understand the population structure of animals. Raccoon dogs (Nyctereutes procyonoides) in South Korea play a key role in the maintenance of food web stability and possess genetic compositions that are unique compared to those in other areas. However, wild raccoon dogs play another role as the main host of various infectious diseases. To establish long-term strategies for disease management, we investigated the genetic structure and possible geographic barriers that influence the raccoon dog population in South Korea by analyzing 16 microsatellite loci. The present study showed that mountains were the major factors responsible for genetic structuring, along with distance. We proposed potential management units (MUs) for raccoon dogs based on the genetic structuring and gene-flow barrier data obtained in this study. Four MUs were suggested for the Korean raccoon dog population (Northern, Central, Southwestern, and Southeastern). The Korean raccoon dog population structure determined in this study and the proposed MUs will be helpful to establish pragmatic strategies for managing Korean raccoon dog population and for preventing the transmission of infectious diseases.

We investigated the species diversity and phylogeography of the Northeast Asian brown frogs allied to Rana dybowskii (the R. dybowskii species complex: R. dybowskii, R. pirica, and R. uenoi) using four mitochondrial and three nuclear loci. Phylogenetic analyses confirmed the existence of three distinct species in this complex; using extensive molecular data, we confirm the validity of Rana uenoi recognized as a distinct species, and infer R. dybowskii and R. pirica to be sister species. Also, we included populations from previously unsampled regions in Northeast China, and identified them to be R. dybowskii. While many species in Northeast Asia diverged due to Pleistocene glaciation, divergence-dating analyses inferred older, Miocene speciation in the R. dybowskii species complex. Ancestral area reconstruction identified the orogenic movement of the Changbai Mountain Range and the opening of the Sea of Japan/East Sea being major events influencing allopatric speciation.

Accurate taxonomic classification of wildlife species is crucial for guiding biological research and for developing effective management and conservation programs. The taxonomic status of Eurasian badgers from South Korea remains poorly resolved. Here we assessed the phylogenetic relationships and genetic variation of Eurasia badgers using partial mitochondrial fragments to elucidate the evolutionary history and taxonomic status of badgers from the Korean Peninsula. Forty-eight unique haplotypes from 125 individuals were observed. Phylogenetic reconstructions and reduced median networks indicate that Eurasian badgers consisted of four geographic clades (Japan, Eastern Eurasia, Western Eurasia, and Caucasus) with a relatively weak split observed within Eastern Eurasia. Estimated divergence time between the Japanese and Eastern Eurasian clades, including the Korean population, was 467,100 years (69,200–1,085,500 years). The results of this study support the hypothesis that the Japanese badger migrated from the Eurasian continent over the Korea-Japan land bridge and that the Korean Peninsula was an important refugia during the Pleistocene. Our study confirmed that the South Korean badger, Meles meles , belongs to the Eastern Eurasian clade. Based on these results and those of previous studies, we recommend that the scientific name of the Korean badger be changed from M. meles to Meles leucurus (Asian badger).

Orthohantaviruses are negative-sense, single-stranded RNA viruses harbored by rodents, shrews, moles, and bats. Of the shrew-borne orthohantaviruses in the Republic of Korea (ROK), Jeju orthohantavirus (Jeju virus, JJUV) was found on Jeju Island. This small-scale epidemiologic survey investigated the geographic distribution and molecular phylogeny of JJUV in the ROK. In 32 trapping sites, tissues of 84 Crocidura shantungensis were analyzed for JJUV RNA. JJUV RNA was detected in seven (8.3%) shrews captured on the Korean peninsula. The molecular epidemiologic survey demonstrated the prevalence of JJUV by geographic distribution. The RNA loads of JJUV were evaluated in various tissues. Entire coding sequences of tripartite genomes were recovered from two JJUV strains on the mainland. Phylogenetic relationships of the JJUV revealed a distinct geographic lineage of mainland strains from the strains on Jeju Island. This study sheds light on the molecular epidemiology, phylogeographic diversity, and virus-host co-divergence of JJUV, ROK.

The Korean Peninsula is a small but unique area showing great endemic Hynobius diversity with H. quelpaertensis, H. yangi, H. unisacculus and three species candidates (HC1, HC3 and HC4). H. quelpaertensis is distributed in the southern part and in Jeju Island, while the remaining species have extremely narrow distributions. To examine the genetic structure of H. quelpaertensis and the phylogenetic placement in Hynobius. Three mitochondrial and six microsatellite loci were genotyped for 204 Hynobius quelpaertensis, three H. leechii, three H. yangi, three HC1, two H. unisacculus, three HC3, three HC4 and ten Japanses H. lichenatus. A high level of mitochondrial diversity was found in H. quelpaertensis. Our mitochondrial data showed evidence of a historical link between inland and Jeju Island despite the signature of founder effect likely experienced by the early island populations. However, our microsatellite analysis showed the fairly clear signature of isolation history between in- and island populations. Upon phylogenetic analysis, H. quelpaertensis, H. unisacculus and HC1 formed a cluster, whereas H. yangi belonged to a separate cluster. HC3 and HC4 were clustered with either H. quelpaertensis or H. yangi depending on the locus used. Our results show at least partially the historical imprints engraved by dispersal of Korean endemic Hynobius during Pleistocene, potentially providing a fundamental basis in determining the conservation units and finding management strategies for these species.

Current understanding of phylogeographical structure and genetic diversity of Siberian roe deer remains limited mainly due to small sample size and/or low geographical coverage in previous studies. Published data suggest at least two phylogroups: western (Ural Mountains and Western Siberia) and eastern (east from lake Baikal, including the Korean peninsula), but their phylogenetic relationship remains unclear. Combined sequences of cytochrome b (1140 bp) and the mtDNA control region (963 bp) were analyzed from 219 Siberian roe deer from 12 locations in Russia, Mongolia, and South Korea, which cover a large part of its range, to assess genetic diversity and phylogeographical status. Special emphasis was placed on the demographic history and genetic features of central, peripheral, and isolated populations. Results of median-joining network and phylogenetic tree analyses indicate that Siberian roe deer from the Urals to the Pacific Ocean are genetically diverse and that geographical distribution and composition of haplogroups coincide with previously described ranges of the subspecies Capreolus pygargus pygargus and Capreolus pygargus tianschanicus. We found that peripheral populations in the northwestern parts of the species range (Urals), as well as the isolated population from Jeju Island, are genetically distinct from those in the core part of the range, both in terms of genetic diversity and quantitative composition of haplogroups. We also found that northwestern (Urals) and northern (Yakutia) peripheral populations share the same haplogroup and fall into the same phylogenetic clade with the isolated population from Jeju Island. This finding sheds light on the taxonomic status of the Jeju Island population and leads to hypotheses about the discordance of morphological and genetic evolution in isolated populations and specific genetic features of peripheral populations.

The leopard, Panthera pardus, is a threatened species in its range throughout the world. Although, historically, the Korean Peninsula had a high population density of leopards, they were extirpated from South Korea by 1970, leaving almost no genetic specimens. Traditionally, Korean leopards are classified as Panthera pardus orientalis; however, their classification is based only on locality and morphology. Therefore, there is a need for genetic studies to identify the phylogenetic status of Korean leopards at the subspecies level. Presently, no extant wild specimen is available from South Korea; therefore, we extracted genetic material from the old skin of a leopard captured in Jirisan, South Korea in the 1930s and conducted the first phylogenetic study of the South Korean leopard. A total of 726 bp of mitochondrial DNA, including segments of the NADH5 and control region, were amplified by PCR. A phylogenetic analysis of the fragment, along with sequences of nine leopard subspecies from GenBank revealed that the extinct South Korean leopard belonged to the Asian leopard group and in the same clade as the Amur leopard (Panthera pardus orientalis). Thus, the leopard that inhabited South Korea in the past was of the same subspecies as the Amur leopard population currently inhabiting the transboundary region of Russia, China, and North Korea. These results emphasize the importance of conserving the endangered wild Amur leopard population (estimated to be about 60–80 individuals) in Russia and China, for future restoration of leopards in the Korean Peninsula.

The raccoon dog (Nyctereutes procyonoides) is endemic to East Asia but has been introduced in Europe. Its high adaptability enabled its rapid colonization of European countries, where population growth has been raising concerns regarding ecosystem disturbance and the spread of zoonotic diseases. The genetic diversity and structure of endemic, source, and introduced populations from seven locations across South Korea, China, Russian Far East, Finland (spread to Finland after introduction to European part of Russia from Russian Far East), Vietnam, and Japan (Honshu and Hokkaido) were examined based on 16 microsatellite loci. Two major and significantly different (FST = 0.236) genetic clusters were found: continental (South Korean, Chinese, Russian, Finnish, and Vietnamese) and island (Japanese) populations. The continental raccoon dog population comprises three subpopulations (Chinese_Russian_Finnish, South Korean, and Vietnamese) and the Japanese population consists of Honshu and Hokkaido subpopulations. The genetic diversity and geographic structure of raccoon dogs in East Asia has been influenced by natural barriers to gene flow and reveals a typical central-marginal trend in genetic diversity (continental vs. island, and central vs. marginal or source vs. introduced within continental populations). The detected differences between continental and island populations agree with those reported in previous studies that considered these populations as different species.

Hynobius unisacculus is a hynobiid salamander species found only in a limited area at the southernmost part of the Korean Peninsula. Here, we characterized the complete mitochondrial genome of this species that was used to identify the phylogenetic relationship with other Hynobius species. The whole sequence was 16,411 bp and included 13 protein-coding genes, 2 ribosomal RNA genes, and 22 transfer RNA genes. The gene arrangement was completely identical to those observed in other Hynobius species. Upon robust phylogenetic tree reconstructed based on 13 protein-coding genes, Hynobius species living on the Korean Peninsula showed close phylogenetic affinity; H. unisacculus and H. quelpaertensis formed a cluster that was the sister to the cluster of H. leechii and H. yangi.

Aim Pelophylax frogs in East Asia provide an opportunity to explore the impact of glacial cycling on demographic and genetic dynamics, because it has been suggested that they experienced distribution shifts and subsequent mtDNA introgression from Pelophylax plancyi to Pelophylax nigromaculatus in association with climatic oscillations. However, their evolutionary history, including the pattern of introgression, is incompletely understood. We used phylogenetic analyses based on multiple markers to address their evolutionary history, and palaeodistribution modelling to test whether the predicted distribution can explain the pattern of introgression suggested by molecular phylogenetics. Location East Asia, including far-eastern Russia, mainland China, the Japanese archipelago, the Korean Peninsula and Taiwan. Methods Ninety-nine samples of the Pelophylax nigromaculatus species complex were collected from 75 localities throughout the distributional range of each species. Phylogenetic analyses were performed with the mitochondrial cytochrome b (cytb) gene and six nuclear genes using two European Pelophylax frogs as outgroups. Additionally, palaeodistributions of P. nigromaculatus and P. plancyi were predicted using bioclimatic variables. Results Differences were observed between the mitochondrial and nuclear DNA trees in the phylogenetic position of P. plancyi. Similar phylogenetic discordance was also observed within P. nigromaculatus. Distribution modelling suggested that a considerable distributional shift of P. nigromaculatus occurred in association with glacial cycling and that the distributional range of P. plancyi was relatively narrow during glacial periods. Main conclusions The analyses detected five historical mtDNA introgression events and determined their directions, none of which had been deduced in previous studies. In association with glacial cycling, the distributional ranges of P. nigromaculatus and P. plancyi appear to have shifted repeatedly, resulting in multiple complex introgressions. By combining phylogenetic analyses with palaeodistribution modelling, our results supported the hypothesis that the history of mtDNA introgression among Pelophylax frogs was shaped by glacial cycling.

Morphological characteristics reflect geographical variation resulting from adaptation to varying environmental conditions. Carnivore species distributed over a wide geographical range generally have highly polymorphic morphological variation. The raccoon dog (Nyctereutes procyonoides) has a longitudinal distribution restricted to East Asia and the northern Indochina Peninsula. Its unique geographical range makes it an appropriate model to examine how morphological differences are influenced by geography. To demonstrate morphological evolution of Russian, Chinese, Korean and Japanese raccoon dogs predicted by geographical differences, we tested the island rule and Bergmann's rule. We compared craniodental variation among populations and examined morphological implications for intraspecific taxonomic status. Insular raccoon dogs possessed substantially smaller body size than those from the mainland. Moreover, different island effects among Japanese islands were demonstrated by markedly larger occipital condyle breath in the Hokkaido population. Larger skull size in Russian and Hokkaido raccoon dogs could be explained by Bergmann's rule. Based on previous chromosomal and molecular studies and results of our morphological analyses, we suggest Japanese raccoon dogs are a distinct species from the mainland N. procyonoides.

The long-tailed goral, Naemorhedus caudatus, is an internationally endangered species. This species is distributed throughout Northeastern Asia including Northeastern China, the Russian Far East and the Korean peninsula. The population size of long-tailed gorals is currently decreasing in South Korea, and thus effective conservation of the animal is urgently needed. Although the evolution and phylogeny of this animal have been studied, population genetic studies are needed to design effective conservation and management strategies. To evaluate the present status of genetic diversity and genetic structure of long-tailed gorals in South Korea, we investigated genetic variability among 68 goral individuals from different regions, including 11 captive zoo animals, at 12 microsatellite loci. The level of genetic diversity was moderate in wild goral populations, but lower in the captive group. The goral population from the lower northeast region of South Korea was distinct from the upper northeast population, probably due to the natural climatic and geographic conditions. The genetic characteristics of the captive group were more similar to those of the upper northeast population than the lower northeast, confirming that the zoo animals originated in the Seorak Mountain range. Direct translocations between the upper and lower northeast populations are not currently recommended considering the natural population structure and the moderate levels of genetic diversity in the two populations. This study highlights the importance of genetic information in designing effective conservation strategies and translocations of endangered animals, including the Korean goral.

Onychodactylus koreanus, a hynobiid salamander species endemic to the Korean Peninsula, can be regarded as a strict ecological specialist, probably vulnerable to anthropogenic environmental modifications and climate change. We used mitochondrial cytochrome b gene to analyze the genetic diversity and phylogenetic structure of O. koreanus from 19 populations collected in an attempt to cover its major distribution within South Korea. A total of 76 haplotypes of O. koreanus obtained in our analyses could be subdivided into three phylogenetic clades, KR, NE and SE. Clade KR haplotypes occur in most of the regions throughout the Korean Peninsula with four distinct subclades (KR I-IV). Clade NE and SE haplotypes were only observed in two populations YY and YS, respectively. Haplotype sharing was scarce even among populations in geographical proximity, and most of the populations were represented by a single clade or subclade, indicating the low level of gene flow among populations. O. koreanus likely originated from the historical southward dispersal of its ancestral lineages following divergence from Chinese O. zhaoermii that was recovered as the sister of O. koreanus in our phylogenetic analysis. Our results have critical implications for the taxonomic status of O. koreanus and its long-term management plan.

To investigate the phylogenetic relationships of the genus Laticauda to related higher taxa, we compared the sequences of four mitochondrial genes (12S rRNA, 16S rRNA, ND4, Cytb) from three Laticauda species (L. colubrina, L. laticaudata, and L. semifasciata) with those of 55 Asian and Australo-Melanesian elapid species. We also characterized the complete mitogenomes of the three Laticauda species and compared the sequences of 13 mitochondrial genes from Laticauda species with five terrestrial elapid and one viperid species to estimate phylogenetic relationships and divergence times. Our results showed that the genus Laticauda is paraphyletic to terrestrial elapids and diverged from the Asian elapids approximately 16.23 Mya. The mitogenomes of the three Laticauda species commonly encoded 13 proteins, 22 tRNAs, 12S and 16S rRNAs and two control regions and ranged from 17,170 and 17,450 bp in size. The L. colubrina mitogenome was more similar to that of L. laticaudata than that of L. semifasciata. The divergence time among the three Laticauda clades was estimated at 8–10 Mya, and a close phylogenetic relationship between L. colubrina and L. laticaudata was found. Our results contribute to our understanding of the evolutionary history of sea kraits.

The water deer (Hydropotes inermis) is one of the rarest species of deer in the family Cervidae. Only two subspecies exist in East Asia, and few studies have examined the genetic characteristics of the species. Here, we investigated the genetic diversity, phylogeny and population differentiation of the Korean subspecies (H. inermis argyropus). Seventeen mitochondrial D-loop haplotypes (822 bp) were detected and analyzed from 107 individual samples, together with a Chinese subspecies (H. inermis inermis) haplotype. The genetic diversity of the Korean subspecies is lower (π = 0.756%, h = 0.867) than that of the Chinese subspecies estimated in a previous study. This low genetic diversity may result from historical anthropogenic disturbances and/or a founder effect during the glacial period. The phylogenetic tree and median-joining network showed no location-specific distribution of D-loop haplotypes, but revealed two major lineages, A and B, of water deer. The A and B lineages were separated from each other at the beginning of the Pleistocene era (2.1-1.3 million years ago), with a genetic divergence of 1.332 ± 0.340%. The genetic divergence within lineages A and B was 0.525 ± 0.167% and 0.264 ± 0.113%, respectively. This suggests that climate change affected the division of the two lineages. Water deer sampled from the three Korean regions (26 locations) were slightly distinct in their genetic structure (AMOVA: F(ST) = 0.28416, P < 0.00001; Φ(ST) = 0.19239, P < 0.00001). Such slight population differentiation may be derived from differential dispersal ability in males and females. The use of genetic markers, such as nuclear microsatellite and Y-linked DNA markers, and samples collected from various localities in East Asia should improve our understanding of the water deer's genetic characteristics.

The hair microstructures of Korean terrestrial mammals from 23 species (22 wild and one domestic) were analyzed using light and scanning electron microscopy (SEM) to construct a hair identification key. The hairs were examined using the medulla structures and cuticular scales of guard hairs from the dorsal regions of mature adult animals. All cuticular scale structures in the hair of Rodentia, Lagomorpha, Carnivora and Insectivora showed the petal pattern, and those of Artiodactyla and Chiroptera showed the wave pattern and coronal pattern, respectively. Rodentia, Lagomorpha and Carnivora showed multicellular, and Insectivora and Artiodactyla showed unicellular regular, mesh or columnar in the medulla structures, respectively. Chiroptera did not show the medulla structures in their hair. We found that it is possible to distinguish between species and order based on general appearance, medulla structures and cuticular scales. Thus, we constructed a hair identification key with morphological characteristics from each species. This study suggests that hair identification keys could be useful in fields, such as forensic science, food safety and foraging ecology.

Population declines and extinctions of amphibians have been attributed to the chytrid fungus Batrachochytrium dendrobatidis (Bd), especially one globally emerging recombinant lineage ('Bd-GPL'). We used PCR assays that target the ribosomal internal transcribed spacer region (ITS) of Bd to determine the prevalence and genetic diversity of Bd in South Korea, where Bd is widely distributed but is not known to cause morbidity or mortality in wild populations. We isolated Korean Bd strains from native amphibians with low infection loads and compared them to known worldwide Bd strains using 19 polymorphic SNP and microsatellite loci. Bd prevalence ranged between 12.5 and 48.0%, in 11 of 17 native Korean species, and 24.7% in the introduced bullfrog Lithobates catesbeianus. Based on ITS sequence variation, 47 of the 50 identified Korean haplotypes formed a group closely associated with a native Brazilian Bd lineage, separated from the Bd-GPL lineage. However, multilocus genotyping of three Korean Bd isolates revealed strong divergence from both Bd-GPL and the native Brazilian Bd lineages. Thus, the ITS region resolves genotypes that diverge from Bd-GPL but otherwise generates ambiguous phylogenies. Our results point to the presence of highly diversified endemic strains of Bd across Asian amphibian species. The rarity of Bd-GPL-associated haplotypes suggests that either this lineage was introduced into Korea only recently or Bd-GPL has been outcompeted by native Bd strains. Our results highlight the need to consider possible complex interactions among native Bd lineages, Bd-GPL and their associated amphibian hosts when assessing the spread and impact of Bd-GPL on worldwide amphibian populations.

The Korean Peninsula is a problematic place for tracing the evolutionary history of many East Asian species because of its location on the eastern edge of the Eurasian continent. This peninsula probably experienced peripheral isolation as one of several possible Pleistocene refugia. Historical population fluctuations and peripatric speciation of vertebrates in the Korean Peninsula are poorly understood. As an endemic species in East Asia, the raccoon dog Nyctereutes procyonoides is an appropriate model for describing the evolutionary history of mammal species in the Korean Peninsula. Therefore, we used mitochondrial cytochrome b sequences of raccoon dogs from Korea, Russia, China, Vietnam and Japan to test four hypotheses: (1) during glacial periods, a single contiguous refugium may have existed in north-eastern Asia that permitted genetic exchange between raccoon dogs from Korea and Japan; (2) the presence of one large refugium did not permit gene flow between raccoon dogs in Korea and Japan; (3) several refugia existed on the north-east Asian mainland, one located in the southern part of the Korean Peninsula, with some population movement to Japan; (4) the presence of several refugia, but no gene flow between the raccoon dogs of Korea and Japan. Our results support the last hypothesis. Haplotype distributions indicate postglacial expansion of raccoon dogs in the Korean Peninsula. Conspicuous genetic differences between Japanese and continental populations might be the result of limited gene flow after geographical isolation. This phylogeographical pattern shows the effect of peripheral isolation in the Korean Peninsula, the southernmost refugium for raccoon dogs.

The collared scops owl that occurs in Korea is a protected species but its exact specific status has been questioned. To resolve the species status, a molecular phylogenetic analysis was conducted using two fragments of mitochondrial DNA, cytochrome b (cyt b, 891 bp) and NADH dehydrogenase subunit 2 (ND2, 627 bp) genes. Phylogenetic trees of cyt b revealed that all Korean specimens formed a monophyletic group with Japanese scops owl Otus semitorques with very low sequence divergence (d=0.008). We obtained a similar ND2 tree as well (d=0.003); however, the genetic distance between Korean individuals and O. lempiji from GenBank (AJ004026-7, EU348987, and EU601036) was very high and sufficient enough to separate them as species (cyt b, d=0.118; ND2, d=0.113). We also found that Korean species showed high differentiation from O. bakkamoena (AJ004018-20 and EU601034; cyt b, d=0.106; ND2, d=0.113) and O. lettia (EU601109 and EU601033, cyt b, d=0.110; ND2, d=0.117) as well. Therefore, we suggest that the Korean collared scops owl should be designated as Otus semitorques.

The raccoon dog (Nyctereutes procyonoides) is distributed from southeastern Siberia to northern Vietnam, including Korea and Japan, as well as Europe. In Korea, most of its predators and competitors are extinct, which has resulted in rapid growth of the raccoon dog population. This population increase has raised concerns about its role in the ecosystem and the zoonotic transfer of various contagious diseases, and thus an effective method of raccoon dog population control in Korea is required. To investigate the genetic diversity and structure of raccoon dog populations, 12 polymorphic microsatellite loci were identified and characterized. These novel microsatellite markers were employed to obtain basic population genetic parameters for 104 N. procyonoides specimens from five locations in South Korea. The mean allele number of 12 loci across samples was 8.7, and the number of alleles per locus ranged 2-13. Mean expected and observed heterozygosities were 0.723 and 0.619, respectively. Genetic differentiation, estimated by pairwise FST, was significant for all population pairs excepting Seoul/Gyeonggi and Gangwon pair, with a moderate level of genetic differentiation for all the population pairs (mean FST = 0.054), but little differentiation between Seoul/Gyeonggi and Gangwon (FST = 0.024). Bayesian-based clustering analysis predicted that Korean raccoon dog population is composed of four distinct genetic subpopulations. These genetic information and structure of raccoon dog will be very useful to prevent spreading infectious diseases.

We describe a new species of lentic-breeding Hynobius salamander from the Naro Islands, near the village of Bongrae-myeon, Goheung-gun, Jeollanam-do, South Korea, on the basis of results of morphological, ecological and genetic analyses. Hynobius unisacculus sp. nov. is distinguished from its congeners by a combination of the following morphological attributes: (1) comparatively small size (adult SVL up to 61 mm; range 38.3–60.3 mm in males and 37.5–59.9 mm in females); (2) relatively slender short limbs; tips of fore- and hindlimbs adpressed on body never meeting, but separated by a large gap (gap of -3.0 to -1.5 costal folds in males and -3.5 to -1.5 in females); (3) comparatively short tail (TL/SVL ratio in adult males varying from 0.54–0.98, in adult females from 0.55 to 0.89), tail flattened and with a low dorsal fin extending to the posterior one-third of tail length; (4) usually 11 (occasionally 12) costal grooves; (5) in adults, dark brown dorsum with indistinct bronze or dark copper spots, lighter greyish-white or pinkish belly; (6) well developed fifth toe; (7) comparatively shallow vomerine tooth series with 13–23 vomerine teeth; (8) small, pigmented ova, located in one, occasionally two, strings in a small, curved egg sac with folded envelope, lacking distinct mucous stalks or whiptail-like structures on both ends. The molecular differentiation among Korean Hynobius is high; Hynobius unisacculus sp. nov. is genetically highly divergent from the morphologically similar H. leechii , H. yangi and H. quelpaertensis : pairwise distances are 9.7%, 9.1% and 8.0% of sequence divergence at the COI mtDNA gene respectively, and 10.9%, 10.9% and 9.4% of sequence divergence at the cyt b mtDNA gene, respectively. At present, the new species is known from coastal areas and offshore islands in southeastern part of Jeollanam-do in South Korea. We suggest the species should be considered as Vulnerable (Vu2a) in accordance with IUCN’s Red List categories. Our study supports the presence of undiagnosed taxonomic diversity among Korean Hynobius .

The Korean hare, Lepus coreanus, is an important mammal in ecosystem food chains, and is distributed across the entire Korean peninsula and northeastern China. Polymorphic microsatellite loci were developed using the biotin-enrichment technique for use in population genetics studies. Five trinucleotide and four dinucleotide microsatellite loci were selected and tested on 22 Korean hare specimens collected from Gangwon Province and Gyeongsangbuk Province in South Korea. The number of alleles across the two sampling regions ranged from three to nine with a mean of 6.1. Mean observed and expected heterozygosities and polymorphic information content were 0.540, 0.627 and 0.579, respectively. Only one locus, Lc06, showed departure from Hardy-Weinberg equilibrium after applying the Bonferroni correction. Four microsatellites, Lc01, Lc03, Lc12, and Lc19, satisfied the criteria to serve as a core set of markers recommended for population genetics studies. These new microsatellite markers will be widely a...

Speciation remains a fundamental issue in biology. Herein, we report an investigation into speciation in the Rana chensinensis species complex using DNA sequence data from one mitochondrial and five nuclear genes. A phylogenetic analysis of the data revealed four major clades in the complex, and each of them was found to likely represent a species, including one cryptic species. Ecological niche models were generated from 19 climatic variables for three of the four major clades, which were represented by widespread sampling, including R. chensinensis, Rana kukunoris and the potential cryptic species. Each clade is associated with a unique ecological unit, and this indicates that ecological divergence probably drove speciation. Ecological divergence is likely related to the late Cenozoic orogenesis of the Qinghai-Tibetan Plateau. In addition, gene flow between species was detected but only in peripheral portions of the ranges of the four major clades, thus likely had little influence on the speciation processes. Discordances between mitochondrial and nuclear genes were also found; the nominal species, R. chensinensis, contains multiple maternal clades, suggesting potential mitochondrial introgression between R. chensinensis and R. kukunoris.

We examined sexual dimorphism in the craniodental traits of the raccoon dog Nyctereutes procyonoides from South Korea. Univariate comparisons of skull (cranium and mandible) and dental measurements revealed a small extent of sexual dimorphism in some measurements. The most indicative dimorphic measurements were the breadths of the upper and lower canines which were around 8% larger in male specimens on average. On the other hand, multivariate analyses using only skull traits showed slightly a clearer separation between sexes than those using only dental ones. This discrepancy may be derived from a higher variability in dental traits than in those of the skull. In conclusion, sexual dimorphism within N. procyonoides of South Korea is present, but was not so pronounced as for other local populations. However, measurements showing significant sexual dimorphism varied between different localities. This suggests that the selective forces acting upon craniodental morphology of each sex vary between populations of the species.

The Eurasian otter, Lutra lutra, is a well known endangered species in South Korea that experienced rapid population decreases through poaching and industrialization until the 1980’s. To evaluate the genetic diversity and the existing number of otters recently found in Daegu City, 81 fecal samples collected from the Gumho River and Shincheon stream were subjected to DNA extraction for sex determination and genetic analysis using nuclear genetic markers. Individual identification and relatedness between individuals were detemined by genotypic data using twelve microsatellite loci, and sex identification was also determined based on sequence variation of the zinc finger protein gene on sex chromosomes. Our results showed that at least seven otter individuals were identified and the kinship relationships of seven individual pairs were determined. It was concluded that otters distributed widely in Daegu City have moderate levels of genetic diversity, and close monitoring of the small-sized otter population is necessary to promote successful settlement of the otters in the area.

The main goals of this study were to isolate microsatellites markers of Korean water deer (Hydropotes inermis argyropus) and understand the genetic status of the species in South Korea. Twelve new microsatellite loci were isolated and characterized to establish basic population genetic parameters for 45 H. i. argyropus specimens in South Korea. There were no significant regional or genetic structure differences between the mid-eastern and southwestern populations in South Korea according to the population genetic analyses. The number of alleles per locus ranged from two to 13 with an average of 6.08. Mean expected (H E) and observed heterozygosity (H O) were 0.622 and 0.533, respectively. Microsatellite variability was also not significant between the two regions (F ST=0.012). These new markers should facilitate the future population genetics studies of Korean water deer and other closely related species.

Siberian chipmunks, Tamias sibiricus, are one of several popular companion animals found in the pet shops of South Korea. At present, however, there have been no studies done in South Korea examining their origin even though they could be potential carriers of zoonotic diseases, and are a species of concern for efficient conservation and management strategies. Sequences of the mitochondrial cytochrome b gene (1140 bp) were determined to investigate the origin of Siberian chipmunks sold in four South Korean pet shops through comparison with sequence data from animals of known locality. Nine Siberian chipmunks were collected from pet shops in South Korea, which resulted in nine haplotypes. One (AR) of these coincided with the haplotype previously described. Phylogenetic and network analyses using 53 haplotypes including 45 haplotypes from GenBank showed three phylogenetic groups in South Korea, almost concordant to locality, designated as northern, central, and southern parts as described in a previous study. Of the nine individuals examined from the pet shops, eight were clustered into the northern phylogroup but one (cgrb9153) was grouped with the southern phylogroup, implying that at least the Siberian chipmunks examined in this study did not originate from other countries. It is likely that most individuals sold in the pet shops of Seoul were caught in the wild in Gyeonggi-do and Gangwon-do, or are maternal descendants of captive-bred individuals originating from the northern part of South Korea. It is recommended that conservation and management units of Korean chipmunks should be examined in further detail.

We studied and compared the age structure, body size, and growth rates of field populations of two Korean salamander species (Hynobius yangi and Hynobius quelpaertensis) to elucidate important aspects of basic population dynamics of these two endemic Hynobius species. In both populations, females were sexually mature at three years of age, while H. yangi and H. quelpaertensis males matured at two and three years of age, respectively. Both males and females of H. yangi and H. quelpaertensis attained a maximum age of 11 years and 10 years, respectively. In both species, the snout-vent length (SVL) and body weight (BW) of the females were greater than those of the males. The SVL, BW, and asymptotic SVL of both male and female H. yangi were smaller than those of H. quelpaertensis. The adult growth rates after sexual maturation of male and female H. yangi were lower than those of H. quelpaertensis, possibly resulting in the smaller body size of the former, although overall growth coefficients were not significantly different between the two species. We also compared the age structure and growth rates of three Korean and three Japanese species of Hynobius.

We have developed 10 microsatellite loci from the Korean ratsnake Elaphe schrenckii. Polymorphism of each locus was assessed in 25 unrelated individuals for E. schrenckii from South Korea, 10 E. anomala and 10 E. schrenckii from China, and 10 E. schrenckii from Russia. E. anomala, a closely related species to E. schrenckii in China. The number of alleles per locus varied from 1–9 (mean 4.4) for Korean populations, 2–7 (mean 3.9) for Chinese E. anomala, 1–7 (mean 4.0) for Chinese E. schrenckii, and 1–4 (mean 2.5) for Russian E. schrenckii. The average expected heterozygosity was 0.500, 0.534, 0.487, and 0.338 for E. schrenckii from South Korea, E. anomala from China, E. schrenckii from China, and E. schrenckii from Russia, respectively. The microsatellite markers used in this study may be applied to other Elaphe species for population analysis, eliminating the time-consuming task of finding new loci. These selected markers are useful for assessing population structure, intraspecific variation, and conservation and management of E. schrenckii.

The complete mitochondrial DNA (mtDNA) control region was analyzed from five species of the subfamily Caprinae; Naemorhedus caudatus, N. goral, Capra hircus, Capricornis swinhoei, and Capricornis crispus. Among these species, the control region ranged from 1,096 to 1,212 bp in length. Our results were compatible with the scheme of three domains (ETAS, Central, and CSB) within the control region. A + T < G + C was observed in all the domains. In the Korean gorals, of the 31 variable sites in the whole control region resulting in 15 haplotypes, 27 variable sites were in the ETAS domain. We found two to three tandem repeat in all five species examined in this study, three in N. caudatus and N. goral, two in Capra hircus and C. crispus, and one in C. swinhoei, respectively. All of these repeat units include two short sections of mirror symmetry (TACAT and ATGTA). Short mirror symmetries were well-resolved among five different species, although left domain has high substitution rates. By Kimura’s two parameter method, the genetic distances between the genera Naemorhedus and Capricornis were calculated and divergence time between Naemorhedus and Capricornis may be nearly 2Myr.

On 28 April 2008, a male Himalayan Swiftlet (Aerodramus brevirostris) was captured and examined at a night roost of swallows on Hongdo Island, Jeollanam-do, Korea. This is the first record of this species from Korea confirmed by specimen examination. We describe morphological features and some phylogenetic notes of the Himalayan Swiftlet found.

Chytridiomycosis, a disease that has caused amphibian population declines globally and elevated many species of anurans to endangered or threatened status, has recently been declared an internationally notifiable disease. Batrachochytrium dendrobatidis (Bd), the amphibian chytrid fungus causing this disease, has not been previously reported in Korea or on mainland Asia. Thirty-six frog specimens representing 7 species were collected from the wild in South Korea and examined for Bd using standard PCR. Bd was detected in 14 (38.8%) samples from 3 species (Bufo gargarizans, Hyla japonica, and Rana catesbiana). Skin sections from all 14 PCR-positive frogs were examined using 2 staining techniques: haematoxylin and eosin (H&E) and Bd immunoperoxidase (IPX). In histological sections, zoosporangia were found in 6 frogs, with lower sensitivity for H&E (21%) than for IPX (46%). Intensity of infection, based on histopathology, was low in all frogs. These results confirm that Bd is present in South Korea and, hence, on the Asian mainland. Studies are urgently required to determine the impact of chytridiomycosis on Korean amphibians, and to map the distribution of Bd in Korea and other Asian mainland countries.

A cytogenetic analysis, including the karyotype, C-bands, silver-stained nucleolus organizer regions and genome size, was performed on the recently discovered species, Karsenia koreana, the first plethodontid salamander from Asia. The karyotype consists of 14 pairs of bi-armed chromosomes, with no evidence of heteromorphic sex chromosomes. C-banding reveals a concentration of heterochromatin at the centromeres as well as at interstitial locations. The smallest chromosome (pair number 14) has symmetrical interstitial C-bands in each arm, resembling chromosome no. 14 of North American species of its sister group taxon, supergenus Hydromantes. Acomparative analysis of C-band heterochromatin and silver-stained nucleolus organizer regions of Karsenia and other plethodontid genera reveals that chromosomal evolution may have featured chromosome 'repatterning' within the context of conserved chromosome number and shape in this clade. Genome size is correlated with geographic distribution in plethodontids and appears to have important phenotypic correlates as well. The genome size of Karsenia is relatively large, and resembles that of the geographically closest plethodontids from western North America, especially species of the genus Hydromantes. The biological significance of these cytogenetic characteristics of plethodontid salamanders is discussed within an evolutionary context.

Siberian flying squirrel, an endangered species in South Korea, is distributed through major mountain regions of South Korea. The number of Siberian flying squirrel (Pteromys volans) in South Korea has decreased and their habitats are fragmented and isolated because of anthropogenic activities. So far no molecular genetic data has, however, been available for their conservation and management. To obtain better information concerning genetic diversity and phylogenetic relationships of the Siberian flying squirrel in South Korea, we examined 14 individuals from South Korea, 7 individuals from Russia, and 5 individuals from northeastern China along with previously published 29 haplotypes for 1,140 bp of the mtDNA cytochrome b gene. The 14 new individuals from South Korea had 7 haplotypes which were not observed in the regions of Russia and Hokkaido. The level of genetic diversity (0.616%) in the South Korean population was lower than that in eastern Russia (0.950%). The geographical distribution of ...

Background The roe deer, Capreolus sp., is one of the most widespread meso-mammals of Palearctic distribution, and includes two species, the European roe deer, C. capreolus inhabiting mainly Europe, and the Siberian roe deer, C. pygargus, distributed throughout continental Asia. Although there are a number of genetic studies concerning European roe deer, the Siberian roe deer has been studied less, and none of these studies use microsatellite markers. Natural processes have led to genetic structuring in wild populations. To understand how these factors have affected genetic structure and connectivity of Siberian roe deer, we investigated variability at 12 microsatellite loci for Siberian roe deer from ten localities in Asia. Results Moderate levels of genetic diversity (HE = 0.522 to 0.628) were found in all populations except in Jeju Island, South Korea, where the diversity was lowest (HE = 0.386). Western populations showed relatively low genetic diversity and higher degrees of genetic differentiation compared with eastern populations (mean Ar = 3.54 (east), 2.81 (west), mean FST = 0.122). Bayesian-based clustering analysis revealed the existence of three genetically distinct groups (clusters) for Siberian roe deer, which comprise of the Southeastern group (Mainland Korea, Russian Far East, Trans-Baikal region and Northern part of Mongolia), Northwestern group (Western Siberia and Ural in Russia) and Jeju Island population. Genetic analyses including AMOVA (FRT = 0.200), Barrier and PCA also supported genetic differentiation among regions separated primarily by major mountain ridges, suggesting that mountains played a role in the genetic differentiation of Siberian roe deer. On the other hand, genetic evidence also suggests an ongoing migration that may facilitate genetic admixture at the border areas between two groups. Conclusions Our results reveal an apparent pattern of genetic differentiation among populations inhabiting Asia, showing moderate levels of genetic diversity with an east-west gradient. The results suggest at least three distinct management units of roe deer in continental Asia, although genetic admixture is evident in some border areas. The insights obtained from this study shed light on management of Siberian roe deer in Asia and may be applied in conservation of local populations of Siberian roe deer. Electronic supplementary material The online version of this article (doi:10.1186/s12863-015-0244-6) contains supplementary material, which is available to authorized users.

The Oriental fire-bellied toad (Bombina orientalis) is a commonly used study organism, but knowledge of its evolutionary history is incomplete. We analyze sequence data from four genetic markers (mtDNA genes encoding cytochrome c oxidase subunit I, cytochrome b, and 12S–16S rRNA; nuDNA gene encoding recombination activating gene 2) from 188 individuals across its range in Northeast Asia to elucidate phylogeographic patterns and to identify the historic events that shaped its evolutionary history. Although morphologically similar across its range, B. orientalis exhibits phylogeographic structure, which we infer was shaped by geologic, climatic, and anthropogenic events. Phylogenetic and divergence-dating analyses recover four genetically distinct groups of B. orientalis: Lineage 1—Shandong Province and Beijing (China); Lineage 2—Bukhan Mountain (Korea); Lineage 3—Russia, Northeast China, and northern South Korea; and Lineage 4—South Korea. Lineage 2 was previously unknown. Additionally, we discover an area of secondary contact on the Korean Peninsula, and infer a single dispersal event as the origin of the insular Jeju population. Skyline plots estimate different population histories for the four lineages: Lineages 1 and 2 experienced population decreases, Lineage 3 remained stable, while Lineage 4 experienced a sharp increase during the Holocene. The timing of the population expansion of Lineage 4 coincides with the advent of rice cultivation, which may have facilitated the increase in population size by providing additional breeding habitat.

Perhaps the oldest unresolved debate inconservation genetics is whether geneticvariability matters – in other words, whetherrelatively low average genetic variationcontributes to deficits in individual andpopulation level vigor and fitness. Using astatistically powerful paired sampling designin which each of three pairs of populationsconsisted of one high genetic variability andone low genetic variability population from aparticular subspecies of the pocket gopher,Thomomys bottae, we tested the hypothesisthat individuals from populations with lowergenetic variability have lower growth rates (acommonly used surrogate for fitness) than thosefrom populations with higher variability. Wemeasured genetic variability using averageallozyme heterozygosity and two measures of DNAfingerprint band sharing (Jeffreys 33.15 andMS1 probes). The population rankings of thelevels of genetic variability among the threemeasures were concordant. The least squaresmean growth rate (controlling for sex,subspecies and initial mass) of gophers fromlow variability populations (0.41 ± 0.06g/day, n = 48) was less than half that ofgophers from high variability populations (1.04± 0.07 g/day, n = 45). This result lendscredence to the premise that differences inpopulation level genetic variability havesignificant fitness consequences andunderscores the importance of maintaininggenetic variability in managed populations.

Allozyme analysis for 41 populations of brown frog species, Rana dybowskii, R. huanrenensis, and R. amurensis from Korea and three reference species (Chinese R. chensinensis and Japanese R. dybowskii and R. tsushimensis), were performed to clarify taxonomic status of Korean brown frogs. The level of average genetic differentiation (Nei's D) among local populations of each species in Korea was very low (D0.01 2) and Korean and Japanese R. dybowskii also showed conspecific level of differentiation (D=0.070). Whereas, much larger, discrete genetic differences were detected in the interspecific comparisons (D0.370). In the genetic relationships among five species examined, the 24 chromosome brown frogs (R. dybowskii, R. huanrenensis, and R. chensinensis) did not form a monophyletic group. Rana dybowskii with the chromosome number of 2n=24 was grouped together with R. amurensis with the chromosome number of 2n=26. The hypothesis of reversal change from 24 to 26 in Korean R. amurensis seems to better explain the phylogenetic relationships of east Asian brown frogs than the assumption of parallel reduction in chromosome number from 2n=26 to 24 in R. dybowskii and in the common ancestor of R. huanrenensis and R. chensinensis. The genetic, morphological, and reproductive divergences between Korean R. dybowskii and R. huanrenensis were compared.

In mice and humans, the androgen receptor (AR) gene, located on the X chromosome, is not known to be involved in sex determination. In the Japanese frog Rana rugosa the AR is located on the sex chromosomes (X, Y, Z and W). Phylogenetic analysis shows that the AR on the X chromosome (X-AR) of the Korean R. rugosa is basal and segregates into two clusters: one containing W-AR of Japanese R. rugosa, the other containing Y-AR. AR expression is twice as high in ZZ (male) compared to ZW (female) embryos in which the W-AR is barely expressed. Higher AR-expression may be associated with male sex determination in this species. To examine whether the Z-AR is involved in sex determination in R. rugosa, we produced transgenic (Tg) frogs carrying an exogenous Z-AR. Analysis of ZW Tg frogs revealed development of masculinized gonads or ‘ovotestes’. Expression of CYP17 and Dmrt1, genes known to be activated during normal male gonadal development, were up-regulated in the ZW ovotestis. Testosterone, supplied to the rearing water, completed the female-to-male sex-reversal in the AR-Tg ZW frogs. Here we report that Z-AR is involved in male sex-determination in an amphibian species.