Morphometric variation in the pusillus group of the genus Rhinolophus (Mammalia: Chiroptera: Rhinolophidae) in East Asia

Based on 203 specimens belonging to the Rhinolophus "pusillus group" (Mammalia: Chiroptera: Rhinolophidae), univariate and multivariate morphometric analyses using 19 characters were performed to assess the confused species taxonomy. The results indicated that R. pusillus (including calidus, parcus, and szechuanus) in the continental region and Hainan Island of China and "R. cornutus" in Japan are morphologically divergent species. Rhinolophus cornutus should be further split into R. cornutus (including orii, pumilus, and miyakonis) in the main islands of Japan, the Amami and Okinawa Group of the central Ryukyu Archipelago, and Miyako Group of the southern Ryukyus; and R. perditus and R. imaizumii from the Yaeyama Group in the southern Ryukyus. Rhinolophus monoceros from Taiwan is morphologically more similar to species in Japan than to R. pusillus. In addition to R. pusillus, another form that is morphologically similar to species in Japan was recognized from Langzhong in Sichuan Province; this may represent an undescribed species, and further examination is necessary to determine its taxonomic status. Specimens from Guang'an in Sichuan Province, China, are also different from the others, and are characterized by the smallest skull size. Although further studies are required, these specimens were tentatively identified as R. subbadius.     

A systematic review of the snakes allied to Amphiesma pryeri (Boulenger) (Squamata: Golubridae) in the Ryukyu Archipelago, Japan

External characters, hemipenial morphology, karyotype, and reproductive mode were examined for the three subspecies of the endemic Ryukyu natricine, Amphiesma pryeri . Results revealed that populations of the Yaeyama Group, currently regarded as comprising a subspecies, A. p. ishigakiense , have several unique features such as the brownish ground colour of head, termination of sulcus spermaticus in the crotch between apical lobes of the hemipenis, terminal location of centromeres in the largest microchromosome pair, and the viviparous mode of reproduction. In the other populations, ground colour of head is black or dark greyish tan; sulcus spermaticus terminates near the tip of the left lobe of the hemipenis; the largest microchromosome pair is metacentric; and the reproductive mode is oviparous. Of these, however, the Miyakojima population, which currendy consists solely of the subspecies, A. p. concelarum , differs from the nominotypical subspecies from the central Ryukyus in having only 94–102 subcaudal scales and telocentric W chromosome; A. p. pryeri has 112–130 subcaudal scales and submetacentric W chromosomes. These strongly suggest that the two subspecies have also substantially diverged from each other, constituting distinct evolutionary lineages. We thus accord full-species status to each of pryeri, concelarum and ishigakiense , and revise their definitions on the basis of the present results. A dichotomous key to the Japanese species and subspecies of Amphiesma is provided.     

Pleistocene speciation of freshwater crabs (Crustacea: Potamidae: Geothelphusa) from northern Taiwan and southern Ryukyus, as revealed by phylogenetic relationships

The phylogenetic relationship among freshwater crab species of Geothelphusa from northern Taiwan and the Yaeyama Group of islands (including Iriomote and Ishigaki) in the southern Ryukyus was studied using the mitochondrial genes 16S rRNA and COI. Our results support the hypothesis that speciation of Geothelphusa among these islands was the result of cyclic glaciations and interglaciations during the Pleistocene. Two main clades, one the Taiwan Group (containing several clades, including most Taiwanese Geothelphusa species except Geothelphusa miyazakii but including Geothelphusa minei from Yaeyama), was estimated to be separated from its sister group, the southern Ryukyus-northern Taiwan (SRN) clade (including G. miyazakii, Geothelphusa shokitai, Geothelphusa fulva and G. marginata from northern Taiwan, the Pinnacle Islands [=Diaoyutai Islands or Senkaku Islands] and Yaeyama) at about 5.3 million years ago (mya). G. shokitai was separated from others within the SRN clade at 2.4 mya, but was probably derived from G. miyazakii in northern Taiwan. The ancestor of G. miyazakii is hypothesised to have dispersed from ancestors in Yaeyama and then isolated at 2.0 mya during the Pleistocene interglaciations. This is similar to the speciation of G. minei in Yaeyama at 1.5 mya, except that its ancestors originated from north-eastern Taiwan. Four clades of freshwater crabs are present in the Fushan Botanical Garden, located in the mountainous area of north-eastern Taiwan, which might be due to the historical rearrangements of the drainage and proximity of the various river origins.     

Differentiation of golden-ringed dragonfly Anotogaster sieboldii (Selys, 1854) (Cordulegastridae: Odonata) in the insular East Asia revealed by the mitochondrial gene genealogy with taxonomic implications

Molecular phylogeographical analyses of Anotogaster sieboldii (Selys, 1854) were conducted to reveal the differentiation process of insular populations. The gene genealogy based on 845 bp of the mitochondrial genes (cytochrome oxidase subunit I and subunit II) indicated that A. sieboldii includes two deeply separated lineages. These two major lineages seem to have differentiated in Miocene before the formation of the insular East Asia. One lineage includes three inner clades that correspond to the populations of northern area (the Japanese main islands, Korean Peninsula, Yakushima), Amamioshima and Okinawajima. Populations of Central Ryukyu, including Amamioshima and Okinawajima, might have been divided from the northern populations in early Pleistocene. The other major lineage includes populations of the Yaeyama Group, Taiwan and East China. The former two populations were reconstructed as a reciprocal monophyletic group. Populations of Taiwan and Yaeyama Groups would have been separated from the continental ones in Pleistocene. These two highly divergent lineages should be recognized as distinct species. Furthermore, the mitochondrial lineages revealed six genetically distinct and geographically isolated assemblages: (1) northern populations, (2) Amamioshima, (3) Okinawajima, (4) Yaeyama Group, (5) Taiwan and (6) East China.     

Fine-Scale Genetic Structure and Demographic History in the Miyako Islands of the Ryukyu Archipelago

The Ryukyu Archipelago is located in the southwest of the Japanese islands and is composed of dozens of islands, grouped into the Miyako Islands, Yaeyama Islands, and Okinawa Islands. Based on the results of principal component analysis on genome-wide single-nucleotide polymorphisms, genetic differentiation was observed among the island groups of the Ryukyu Archipelago. However, a detailed population structure analysis of the Ryukyu Archipelago has not yet been completed. We obtained genomic DNA samples from 1,240 individuals living in the Miyako Islands, and we genotyped 665,326 single-nucleotide polymorphisms to infer population history within the Miyako Islands, including Miyakojima, Irabu, and Ikema islands. The haplotype-based analysis showed that populations in the Miyako Islands were divided into three subpopulations located on Miyakojima northeast, Miyakojima southwest, and Irabu/Ikema. The results of haplotype sharing and the D statistics analyses showed that the Irabu/Ikema subpopulation received gene flows different from those of the Miyakojima subpopulations, which may be related with the historically attested immigration during the Gusuku period (900 − 500 BP). A coalescent-based demographic inference suggests that the Irabu/Ikema population firstly split away from the ancestral Ryukyu population about 41 generations ago, followed by a split of the Miyako southwest population from the ancestral Ryukyu population (about 16 generations ago), and the differentiation of the ancestral Ryukyu population into two populations (Miyako northeast and Okinawajima populations) about seven generations ago. Such genetic information is useful for explaining the population history of modern Miyako people and must be taken into account when performing disease association studies.     

Riding the Kuroshio Current: Stepping stone dispersal of the Okinawa tree lizard across the East Asian Island Arc

Aim

Located hundreds of kilometres offshore of continental mainland Asia, the extremely high level of land vertebrate endemism in the East Asian Island Arc provides an excellent opportunity to test hypotheses regarding biogeographic processes and speciation. In this study, we aim to test alternative explanations for lineage diversification (vicariance versus dispersal models), and further develop a temporal framework for diversification in our focal taxon, which is consistent with the known age of these islands. We achieve these tests by investigating the historical biogeography of the Okinawa tree lizard (Japalura polygonata), one of the few widely‐distributed reptiles across this archipelago.

Location

The East Asian Island Arc: (1) Central Ryukyu (Amami and Okinawa groups); (2) Southern Ryukyu (Miyako and Yaeyama groups); (3) Taiwan and adjacent islands.

Methods

A total of 246 tissues were sampled from 10 localities in the Ryukyu archipelago and 17 localities in Taiwan, covering the entire distributional range of this species, including all subspecies. DNA sequences of the mitochondrial cytochrome b, 16S ribosomal RNA, nuclear BACH‐1 and RAG‐1 genes (total: 4,684 bp) were obtained from these samples. We used maximum likelihood and Bayesian methods to infer phylogeny and divergence time, and used a model‐fitting method of biogeographical inference to estimate ancestral range evolution.

Results

Multiple lines of evidence combine to identify a general pattern of dispersal‐mediated diversification northward through the archipelago, following initial dispersal from Taiwan. These included (1) a phylogenetic estimate, revealing a sequential, south‐to‐north branching pattern; (2) ancestral range estimation, inferring multiple overseas dispersals and subsequent colonization of new landmasses; and (3) a reduction in genetic variation observed in successively‐diverging lineages, decreasing from Taiwan northward, towards more remote islands. These results provide strong statistical support for an interpretation of successive bouts of dispersal via the powerful, well‐documented, south‐to‐north Kuroshio Current. Estimation of divergence times suggests that most clades in southern Ryukyu and Taiwan diverged early, giving rise to lineages that have remained isolated, and that more recently‐diverged lineages then colonized northward to subsequently occupy the landmasses of the Central Ryukyu archipelago.

Main conclusions

Our general inference of biogeographic history in Japalura polygonata suggested that this species originated on Taiwan and the Yaeyama group, and arrived at its current distribution in Miyako, Okinawa, Toku and Amami islands by a series of stepping‐stone dispersals, which we report for the first time for a terrestrial vertebrate endemic to this region.     

Phylogeny and phylogeography of the sword-tailed newt,Cynops ensicauda (Amphibia: Caudata), as revealed by nucleotide sequences of mitochondrial DNA

We investigated the phylogenetic relationships and phylogeography among individuals of the endemic newt (Cynops ensicauda) of the Central Ryukyus, Japan based on the mitochondrial cytochrome b gene. The results of phylogenetic analyses showed the presence of remarkable differentiation between assemblages from the Amami and Okinawa Island Groups, supporting the validity of the subspecies C. e. popei described from the latter on morphological grounds. The group of individuals from Okinawa Island Group was further divided into two distinct subgroups unlike the results of previous morphological and allozyme studies. Geographic ranges of these subgroups overlapped in the northern part of Okinawajima Island. The phylogeographic pattern within the Okinawa Island Group suggests an initial division into two geographically isolated population lineages and subsequent secondary sympatry before formation of reproductive isolation. It is also likely that within the Okinawa Island Group emigration occurred from the central and northern parts of Okinawajima Island to its southern part, as well as to several small islets off its western coast. Within the Amami Island Group, recurring restricted gene flow with isolation by distance seems to have occurred around the southwestern region including three islets southwest of Amamioshima Island.     

Phylogeny, taxonomy, and biogeography of the oriental pitvipers of the genus trimeresurus (reptilia: viperidae: crotalinae): a molecular perspective

Based on sequence variation in 806 bp of the mitochondrial 12S rRNA gene, phylogenetic relationships were inferred for 14 species of Trimeresurus (sensu lato) including all East Asian members. Samples analyzed also included representatives of all assemblages of species that are frequently treated as separate genera except for T. mangshanensis, a type species of the recently described monotypic genus Ermia. Results support some previous accounts chiefly from morphological studies, such as distinct divergence of T. wagleri from the remainder, and monophyly of T. mucrosquamatus, T. flavoviridis, T. jerdonii, T. elegans and T. tokarensis. On the other hand, our results negated a putative close affinity of T. monticola and T. okinavensis, and indicated the sister relationship of the latter with T. gracilis. Phylogenetic relationships revealed in this study suggested that the genus Trimeresurus dispersed into the Ryukyu region at least three times, and that T. flavoviridis and T. tokarensis from the central Ryukyus constitute a relict clade.     

Taxonomic study of the Burmoniscus ocellatus complex (Crustacea, Isopoda, Oniscidea) in Japan shows genetic diversification in the southern Ryukyus, southwestern Japan

To clarify the taxonomic status of the Burmoniscus ocellatus complex in Japan, we carried out morphological observations and phylogenetic analyses of specimens collected from Yonagunijima, Iriomotejima, Ishigakijima, and Miyakojima Islands of the southern Ryukyus and from Okinawajima Island of the central Ryukyus in southwestern Japan. Observations of the holotypes of Aphiloscia iriomotensis ( Nunomura, 1986 ), Ap. ishigakiensis ( Nunomura, 1986 ), and Ap. yonakuniensis ( Nunomura, 1986 ), in addition to the specimens newly collected from the five islands, indicated that these specimens belong to the genus Burmoniscus. Analyses of five morphological characters of 268 specimens collected from the five islands showed that the body size of Okinawajima specimens is distinctly smaller than those of the specimens from the southern Ryukyus. The ranges of the five morphological characters tended to overlap among the specimens from Yonagunijima, Iriomotejima, Ishigakijima and Miyakojima Islands; these morphological characters were congruent with those of 6. ocellatus (Verhoeff, 1928). The phylogenetic analyses were based on three regions of mitochondrial DNA-COI, 12S rRNA, and 16S rRNA-and indicated that the specimens collected from the southern Ryukyus constitute a monophyletic group, which is clearly distinct from the clade composed of the Okinawajima specimens. These results strongly suggest that Ap. iriomotensis, Ap. ishigakiensis, and Ap. yonakuniensis are synonymous with B. ocellatus, a species widely distributed in the southern Ryukyus. On the other hand, the species from Okinawajima Island in the central Ryukyus is considered to be an undescribed Burmoniscus species, which is closely related to but clearly distinct from S. ocellatus.     

Relict distribution of Microhyla (Amphibia: Microhylidae) in the Ryukyu Archipelago: High diversity in East Asia maintained by insularization

The Ryukyu Archipelago, located at the southwestern part of Japan, is known as a group of continental islands and harbours many endemic taxa, supposedly reflecting its fairly long isolation from the Eurasian continent, Taiwan and the Japanese main islands. Microhyla okinavensis has been known as an endemic member of the terrestrial fauna of this archipelago. Molecular phylogenetic analyses using samples from nearly all island populations of the species and representative samples of other east Asian congeneric species revealed that M. okinavensis consists of four distinct subclades, of which the Amami, Okinawa and Miyako subclades, though exhibiting distinct genetic differentiations from each other, formed a monophyletic group (clade A). The remaining Yaeyama subclade was exclusively sister to M. mixtura from inland China, forming another monophyletic group (clade B), rendering M. okinavensis in the current definition paraphyletic. These results, as well as estimated dates of divergence from related taxa, indicate that M. okinavensis actually includes more than one distinct species. The results indicate that M. okinavensis and M. mixtura are relict species with disjunct distributions which had been most probably caused by invasion of M. fissipes in intervening areas.     

Phylogenetic divergence of island biotas: Molecular dates,extinction, and “relict” lineages

Island formation is a key driver of biological evolution, and several studies have used geological ages of islands to calibrate rates of DNA change. However, many islands are home to “relict” lineages whose divergence apparently pre‐dates island age. The geologically dynamic New Zealand (NZ) archipelago sits upon the ancient, largely submerged continent Zealandia, and the origin and age of its distinctive biota have long been contentious. While some researchers have interpreted NZ's biota as equivalent to that of a post‐Oligocene island, a recent review of genetic studies identified a sizeable proportion of pre‐Oligocene “relict” lineages, concluding that much of the biota survived an incomplete drowning event. Here, we assemble comparable genetic divergence data sets for two recently formed South Pacific archipelagos (Lord Howe; Chatham Islands) and demonstrate similarly substantial proportions of relict lineages. Similar to the NZ biota, our island reviews provide surprisingly little evidence for major genetic divergence “pulses” associated with island emergence. The dominance of Quaternary divergence estimates in all three biotas may highlight the importance of rapid biological turnover and new arrivals in response to recent climatic and/or geological disturbance and change. We provide a schematic model to help account for discrepancies between expected versus observed divergence‐date distributions for island biotas, incorporating the effects of both molecular dating error and lineage extinction. We conclude that oceanic islands can represent both evolutionary “cradles” and “museums” and that the presence of apparently archaic island lineages does not preclude dispersal origins.     

Unexpectedly complex gradation of coral population structure in the Nansei Islands,Japan

To establish effective locations and sizes of potential protected areas for reef ecosystems, detailed information about source and sink relationships between populations is critical, especially in archipelagic regions. Therefore, we assessed population structure and genetic diversity of Acropora tenuis, one of the dominant stony coral species in the Pacific, using 13 microsatellite markers to investigate 298 colonies from 15 locations across the Nansei Islands in southwestern Japan. Genetic diversity was not significant among sampling locations, even in possibly peripheral locations. In addition, our results showed that there are at least two populations of A. tenuis in the study area. The level of genetic differentiation between these populations was relatively low, but significant between many pairs of sampling locations. Directions of gene flow, which were estimated using a coalescence‐based approach, suggest that gene flow not only occurs from south to north, but also from north to south in various locations. Consequently, the Yaeyama Islands and the Amami Islands are potential northern and southern sources of corals. On the other hand, the Miyako Islands and west central Okinawa Island are potential sink populations. The Kerama Islands and the vicinity of Taketomi Island are potential contact points of genetic subdivision of coral populations in the Nansei Islands. We found that genetic population structure of A. tenuis in the Nansei Islands is more complex than previously thought. These cryptic populations are very important for preserving genetic diversity and should be maintained.     

Quaternary palaeoenvironmental oscillations drove the evolution of the Eurasian Carassius auratus complex (Cypriniformes,Cyprinidae)

Aim We sought to reconstruct the spatio‐temporal genetic diversification in goldfish of the Carassius auratus complex, which is widely distributed in Eurasia, to test whether vicariance events or human‐mediated translocations best explained lineage divergence and genogeographical history. Location East Asia and the Oriental islands including Japan, the Ryukyus and Taiwan, and Europe, including Russia and the Czech Republic. Methods We reconstructed the matrilineal history of Eurasian goldfish using 1876 sequences from the partial mitochondrial DNA control region (426 bp) and 191 complete sequences of cytochrome b (1140 bp) from 67 localities representing most of the range of the species. Divergence times were estimated using a Bayesian Markov chain Monte Carlo approach based either on molecular clock data or on the fossil record. Genetic structure and the historical demography of populations were analysed using partial correlation tests and analyses of molecular variance. Results Three lineages had high levels of regional specificity. Lineages A and B from the main islands of Japan differed greatly from lineage C, which occurred on the mainland, Taiwan and the Ryukyus. Lineages A and B had late Pliocene origins. Six geographically constrained sublineages within lineage C had near‐simultaneous mid‐Pleistocene divergences. Main conclusions Genetic structure in the C. auratus complex appears to have been driven by palaeoenvironmental perturbations rather than human translocations. The disappearance of a land bridge in the Tsushima Strait around 3.0 Ma is responsible for the separation of Japanese and continental lineages; the estimated divergence time is 2.75–2.32 Ma. Fujian, China and Vietnam appear to have provided important refugia for the C. auratus complex during glaciation. After warm, moist summer monsoons intensified during the mid‐Pleistocene, goldfish are likely to have dispersed north‐eastwards to recolonize the Ryukyus via Taiwan and northwards to recolonize mainland China.     

Epidemiological and Ecological Studies of Japanese Encephalitis in Okinawa,Subtropical Area in Japan. II. Prevalence of Japanese Encephalitis Antibody in Residents in Okinawa,Miyako and Ishigaki Islands

During 1989 to 1990, human sera were collected by age groups in Okinawa (the northern, central and southern areas), Miyako and Ishigaki islands and examined for the neutralization (N) antibodies to two strains, Nakayama (vaccine strain) and C307 (Okinawan strain), of Japanese encephalitis (JE) virus. In Okinawa island, the N antibody positive rate to C307 was higher than that to Nakayama, while in Miyako and Ishigaki islands, the positive rate to Nakayama was higher than that to C307, suggesting that JE virus transmission rate was higher in Okinawa than in Miyako and Ishigaki islands. In Okinawa Prefecture, JE vaccine had not been administered to most of residents over 31 years of age at the time of serum collection. In residents over 31 years old, the positive rate to C307 was highest in the north of Okinawa (83.3%) and was lowest in Miyako (26.3%), with the second lowest in Ishigaki (33.3%). The distribution of N antibody titers to C307 gave hyperbolic patterns in the 0–5 age groups in Miyako and Ishigaki, and also in the 31–40, 41–50 age groups in Miyako and the 41–50 age group in Ishigaki, suggesting low rates of natural infection in these 4–5 decades in both islands. In residents of ages subjected to JE vaccine, a characteristic pattern was obtained, in which the curves to Nakayama shifted to higher titers than those to C307, suggesting that the first antigenic stimulation was caused by vaccine, not by natural infection of JE virus.     

Diagnosis and phylogeny of the new world streptocephalidae (Branchiopoda: Anostraca)

We present an updated diagnosis of 13Streptocephalus species of North America. Three new species are included. A key to the species is provided.The phylogeny of the group is discussed on the basis of (1) a systematic approach (Maeda-Martinezet al., 1995; this volume), which considers the entire distal antennal outgrowth, the frontal appendage, and the morphology of the ovaries as essential in defining different genetic lineages or species-groups, and (2) a cladistic analysis. We suggest that of nine monophyletic groups, three are represented in both the Old and New World. Thus, contrary to former disparsalist hypotheses, we argue that the New World species represent relict forms of ancestral groups fragmented by continental drift (vicariance model).     

Population structure of the freshwater flatworm Crenobia alpina (Dana): old lineages and low gene flow

Aim  The freshwater flatworm Crenobia alpina lives almost exclusively in headwaters of mountainous areas and is supposed to be a glacial relict. We examined genetic diversity within and between populations of C. alpina in order to determine the taxonomic status of purported subspecies and to understand large-scale biogeographical patterns of glacial relicts.
Location  Central Europe.
Methods  We analysed mitochondrial DNA sequences and polymorphic allozyme loci of C. alpina populations across its range in central Europe. Sequences were compared using parsimony, minimum evolution and maximum likelihood. Allozymes were analysed using traditional as well as Bayesian estimates of F statistics.
Results  We found considerable divergence between haplotypes. For each of the two lineages occurring throughout central Europe, allozymes showed considerable differentiation between populations and a strong isolation by distance effect. Hence populations are effectively isolated even across rather small spatial scales.
Main conclusions  There is strong evidence that C. alpina is a complex of distinct lineages or cryptic species that date back to the late Miocene. The separation of lineages may be associated with the formation of deep valleys at the end of the Messinian Crisis.     

Sheep mitochondrial DNA variation in European, Caucasian, and Central Asian areas

Three distinct mitochondrial maternal lineages (haplotype Groups A, B, and C) have been found in the domestic sheep. Group B has been observed primarily in European domestic sheep. The European mouflon carries this haplotype group. This could suggest that European mouflon was independently domesticated in Europe, although archaeological evidence supports sheep domestication in the central part of the Fertile Crescent. To investigate this question, we sequenced a highly variable segment of mitochondrial DNA (mtDNA) in 406 unrelated animals from 48 breeds or local varieties. They originated from a wide area spanning northern Europe and the Balkans to the Altay Mountains in south Siberia. The sample included a representative cross-section of sheep breeds from areas close to the postulated Near Eastern domestication center and breeds from more distant northern areas. Four (A, B, C, and D) highly diverged sheep lineages were observed in Caucasus, 3 (A, B and C) in Central Asia, and 2 (A and B) in the eastern fringe of Europe, which included the area north and west of the Black Sea and the Ural Mountains. Only one example of Group D was detected. The other haplotype groups demonstrated signs of population expansion. Sequence variation within the lineages implied Group A to have expanded first. This group was the most frequent type only in Caucasian and Central Asian breeds. Expansion of Group C appeared most recently. The expansion of Group B involving Caucasian sheep took place at nearly the same time as the expansion of Group A. Group B expansion for the eastern European area started approximately 3,000 years after the earliest inferred expansion. An independent European domestication of sheep is unlikely. The distribution of Group A variation as well as other results are compatible with the Near East being the domestication site. Groups C and D may have been introgressed later into a domestic stock, but larger samples are needed to infer their geographical origin. The results suggest that some mitochondrial lineages arrived in northern Europe from the Near East across Russia.     

The Kuroshio Current influences genetic diversity and population genetic structure of a tropical seagrass,Enhalus acoroides

Information on genetic diversity and differentiation of seagrass populations is essential for the conservation of coastal ecosystems. However, little is known about the seagrasses in the Indo‐West Pacific Ocean, where the world's highest diversity of seagrasses occurs. The influence of sea currents on these populations is also unknown. We estimated the genetic diversity and population genetic structure and identified reproductive features in Enhalus acoroides populations from the Yaeyama Islands, Hainan Island and the Philippines. The Philippines are situated at the centre of the E. acoroides range, Yaeyama and Hainan are peripheral populations, and the Yaeyama population is at the northern limit of the species range. The powerful Kuroshio Current flows from the Philippines to Yaeyama. Genetic analyses using nine microsatellite markers indicated that reproduction of E. acoroides is mostly sexual. Clonal diversity does not decrease in northern populations, although genetic diversity does. However, the genetic diversity of the Yaeyama populations is greater than that of the Hainan populations. Significant genetic differentiation among most populations was evident; however, the Yaeyama and north‐east Philippines populations were genetically similar, despite being separated by ~1100 km. An assignment test suggested that recruitment occurs from the north‐east Philippines to Yaeyama. The strong current in this region is probably responsible for the extant genetic diversity and recruitment patterns.     

Phylogeny and biogeography of Rhododendron subsection Pontica, a group with a tertiary relict distribution

Rhododendron subgenus Hymenanthes subsection Pontica is exceptional among Tertiary relict groups in having a high proportion of species (4 of 11) native to SW Eurasia. A phylogeny based on cpDNA matK and trnL-F indicated that multiple Pontica lineages colonised each of SW Eurasia, SE North America, and NE Asia, with little or no speciation within regions thereafter. Therefore, multiple (3-4) Pontica lineages survived the Quaternary in SW Eurasia, in contrast to other Tertiary relict genera. Pontica comprises two major clades, one of which is wholly Eurasian, and paraphyletic with respect to at least some of the remaining 200 species of subgenus Hymenanthes, which are all distributed in SE Asia. The other clade has species from W and SE North America, SW Eurasia, and NE Asia. According to synonymous matK substitution data, the two clades diverged 9-6 million years ago (mya), whereas most divergence within them happened 5-3 mya. Although the phylogeny indicates probable trans-Atlantic migration for one of two America-Eurasia disjunctions in Pontica, the timing supports migration via Beringia for both.