Molecular phylogeny and divergence time of the water penny genus Eubrianax (Coleoptera: Psephenidae) in Japan

The phylogenetic relationships among the Japanese members of the genus Eubrianax (Coleoptera: Psephenidae) were examined using the mitochondrial cytochrome oxidase subunit I (COI) gene and nuclear 28S rRNA gene sequences. Based on the molecular phylogeny as well as morphological features, the species status of Eubrianax brunneicornis Nakane, 1952 was proposed. The phylogenetic analyses recovered monophyly of the previously proposed pellucidus species group with four Japanese species, whereas a single Japanese species of the granicollis group was included in the lineage of the ramicornis group with five Japanese species. The divergence times of the species were estimated by dating the phylogenetic tree against the fossil record and a molecular clock based on the COI gene. The divergence of the Japanese species was inferred to have occurred during the Pliocene epoch.     

Life history and host specificity of the Japanese flea beetles Trachyaphthona sordida and T. nigrita (Coleoptera: Chrysomelidae), potential biological control agents against skunk vine, Paederia foetida (Rubiaceae), in the southeastern parts of the United States and Hawaii

Skunk vine, Paederia foetida (Rubiaceae), is native to Asia and has been recognized as an invasive weedy vine of natural areas in Florida and Hawaii. Two insects, Trachyaphthona sordida and Trachyaphthona nigrita (Coleoptera: Chrysomelidae) from Japan are being considered as potential biological control agents against skunk vine. To gather fundamental information on their biology, we carried out field surveys and laboratory experiments in Kyushu, southern Japan, between 2003 and 2006. We found that T. sordida is commonly distributed in Kyushu and T. nigrita is restricted to the southern parts of Kagoshima Prefecture on the southern part of Kyushu. These species are fundamentally univoltine and adults appear in late April to early July. Trachyaphthona sordida overwinters as mature larvae and T. nigrita as mature larvae or rarely as adults. Larvae of both species feed on fine roots of P. foetida in the field and Serissa foetida (Rubiaceae) under rearing conditions, and they appear to have tribe‐level host specificity in their host range. On the basis of these results, we suggest that both species are suitable as biological control agents.     

Three species of the genus Ilyocryptus (Anomopoda, Branchiopoda) occurring in Japan

Three species of Ilyocryptus, Ilyocryptus sp., I. cuneatus Stifter 1988, and I. spinifer Herrick 1882, are newly recorded from the Japanese islands. The first two species were found separately at two sites in Hokkaido and the third at eight sites in Honshu and Kyushu. Their morphological features are briefly described based on parthenogenetic females. Received: May 21, 2001 / Accepted: July 23, 2001     

Notes on two species of the subgenus Lyrothorax Chaudoir (Coleoptera: Carabidae; genus Pterostichus), Pterostichus amagisanus Tanaka and Ishida and Pterostichus fujitai Tanaka and Ishida

Two species of the subgenus Lyrothorax Chaudoir (Coleoptera: Carabidae; genus Pterostichus ), Pterostichus amagisanus Tanaka and Ishida and Pterostichus fujitai Tanaka and Ishida, were revised based on the male endophallus (inner sac everted from aedeagus). P. amagisanus was newly recorded based on a single male from Kyushu, southwest Japan, far from its known distribution (Honshu; the Fuji-Hakone-Izu volcano area), although additional materials are necessary to confirm this record. Despite a highly disjunct distribution, no conspicuous difference was recognized in either external or genital characters between the materials from Honshu and Kyushu. The nominal species P. fujitai was separated into two species, P. fujitai (Honshu) and Pterostichus eoyoritomus sp. nov. (Shikoku; type locality: Mount Jingayama); these two species have some significant differences in the endophallic structures. Character states in male genitalia suggest a sister relationship between P. eoyoritomus sp. nov. and Pterostichus yoritomus Bates.     

Phylogeography of the leaf beetle Chrysolina virgata in wetlands of Japan inferred from the distribution of mitochondrial haplotypes

The genetic differentiation among populations of the leaf beetle Chrysolina virgata living in wetlands of Japan was studied based on the sequence data of the mitochondrial cytochrome oxidase subunit I gene region (750 bp). Two distinct lineages of mitochondrial haplotypes were found: one (clade A) consisted of 26 haplotypes distributed over the distribution range of C. virgata between north‐east Honshu and Kyushu, whereas the other (clade B) was monotypic and confined to a small region in north‐east Honshu where it coexisted with clade A. Nested clade analysis for these haplotypes suggested that range expansion and following differentiation due to isolation by distance might have resulted in the present distribution pattern of the haplotypes in clade A. We discuss the evolutionary process leading to the occurrence of two distinct haplotype clades in Japan in terms of repeated colonization from the continent and range expansion and contraction during climatic changes.     

Molecular phylogeny of the genus Apatrobus (Coleoptera: Carabidae: Patrobinae) in western Japan

We performed a molecular phylogenetic analysis of the ground beetles Apatrobus (Carabidae), endemic to Japan, using the mitochondrial cytochrome c oxidase subunit I (COI) and the nuclear 28S rRNA (28S) genes. We focused on the species divergence in Kyushu, Shikoku and western Honshu and used 15 of 19 species and three populations with undetermined species in the DNA analysis. The gene trees showed that, of the Apatrobus species studied, A. hayachinensis Nakane from northern Honshu was not included in the monophyletic group of the other Apatrobus species and likely to be of a different genus. Divergence time estimation suggested that Apatrobus species excluding A. hayachinensis diverged 5.2 million years ago and the subsequent divergence of species occurred during the Pliocene and Pleistocene. In each of the main islands, Kyushu, Shikoku and Honshu, two or more distinct lineages occurred and all species had restricted distribution areas, suggesting that ancient dispersal and vicariance among the three main islands resulted in the nested biogeographical pattern of species distribution.     

Cladosiphon umezakii sp. nov. (Ectocarpales, Phaeophyceae) from Japan

The new species Cladosiphon umezakii Ajisaka (Ectocarpales, Phaeophyceae) is described from Japan based on morphology and DNA sequences. The species resembles Cladosiphon okamuranus Tokida in its gross morphology; somewhat slimy, cylindrical, multiaxial and sympodial erect thallus, arising from a small disc‐shaped holdfast, and branching once to twice. However, C. umezakii has considerably longer assimilatory filaments (up to 840 μm long, composed of up to 90 cells) than any known taxa of the genus. The species is a winter to spring annual, growing on lower intertidal to subtidal rocks of more or less exposed sites on the north‐eastern coast of Kyushu and on both the Pacific and the Sea of Japan coasts of Honshu. Specimens from the Sea of Japan coast had both unilocular and plurilocular zoidangia, whereas those from Kyushu and from the Pacific had only unilocular zoidangia. Unilocular zoidangia were formed on the basal part of assimilatory filaments, and plurilocular ones were transformed from the distal part of assimilatory filaments. DNA sequences of the Rubisco‐spacer (rbc‐spacer) region and the nuclear rDNA ITS region (ITS1, 5.8S and ITS2) supported the distinctness of the species.     

Mitochondrial phylogeography of the Japanese pond turtle,Mauremys japonica (Testudines,Geoemydidae)

The phylogeography of the Japanese pond turtle, Mauremys japonica (Temminck and Schlegel, 1835), which is an endemic species in Japan, was studied by analyzing the variation in two mitochondrial DNA sequences, the cytochrome b gene and the control region. These analyses suggest that M. japonica comprises two major groups. The first one was found in the more eastern region, eastern Honshu Island and Shikoku Island, while the second was found in a western region, Kyushu Island and the Chugoku District (the westernmost part of Honshu Island). The boundary between the two groups is located in the Chugoku District. The nucleotide and haplotype diversities were very low, and these low diversities seem to have been caused by a bottleneck in the last glacial age. These results suggest that this species survived the last glacial period in two refugia, one in the central part of Honshu Island and the other one in Kyushu Island. Subsequently, population expansion took place in the postglacial period, and the groups from the two refugia extended their distribution ranges to the present boundary in the Chugoku District which represents a secondary contact zone.     

Genetic structure of <Emphasis Type="Italic">Cerasus jamasakura</Emphasis>, a Japanese flowering cherry,revealed by nuclear SSRs: implications for conservation

The genetic resources of a particular species of flowering cherry, Cerasus jamasakura, have high conservation priority because of its cultural, ecological and economic value in Japan. Therefore, the genetic structures of 12 natural populations of C. jamasakura were assessed using ten nuclear SSR loci. The population differentiation was relatively low (F _ST, 0.043), reflecting long-distance dispersal of seeds by animals and historical human activities. However, a neighbor-joining tree derived from the acquired data, spatial analysis of molecular variance and STRUCTURE analysis revealed that the populations could be divided into two groups: one located on Kyusyu Island and one on Honshu Island. Genetic diversity parameters such as allelic richness and gene diversity were significantly lower in the Kyushu group than the Honshu group. Furthermore, STRUCTURE analysis revealed that the two lineages were admixed in the western part of Honshu Island. Thus, although the phylogeographical structure of the species and hybridization dynamics among related species need to be evaluated in detail using several marker systems, the Kyusyu Island and Honshu Island populations should be considered as different conservation units, and the islands should be regarded as distinct seed transfer zones for C. jamasakura, especially when rapid assessments are required.     

Reinstatement of Myelophycus caespitosus (Scytosiphonaceae,Phaeophyceae) from Japan

Reinstatement of Myelophycus caespitosus Kjellman (Ectocarpales s.l., Phaeophyceae) described from western Japan is proposed based on the comparisons of DNA sequences of Japanese Myelophycus specimens, the type specimen of Chordaria simplex and lectotypified specimen of Myelophycus caespitosus. In the genetic analyses using mitochondrial cox1 and cox3, chloroplast atpB, psbA and rbcL DNA sequences, the specimens morphologically referable to M. simplex formed two distinct clades (clade‐1 and clade‐2) supported by high statistical values. Clade‐1 was distributed on the western coast of Japan (Honshu, Shikoku and Kyushu) from the Kii Peninsula to Tsushima and the Pacific coast of Aomori, northeastern Honshu, and clade‐2 on the Pacific coast of central Honshu from Sanriku to the Kii Peninsula. Based on the cox3 DNA sequences and the location of the type locality at Shimoda, Izu Peninsula, clade‐2 was concluded to correspond to true M. simplex. There were no recognizable differences in the representative morphological features (height and diameter of sporophytes and gametophytes, number of cells comprising cortical, subcortical and medullary layers, thickness of plurilocular gametangia, or length and diameter of unilocular zoidangia) between the specimens included in the two taxa. Analyses of the specimens of the two taxa, including sympatric populations on the Kii Peninsula, using a nuclear genetic marker ocm3 did not suggest any genetic exchanges between the two taxa. On the other hand, cox3 gene sequence of the voucher specimen of Myelophycus caespitosus collected from Goto, Kyushu, western Japan housed in the UPS herbarium was included in clade‐1. In conclusion, we propose the reinstatement of M. caespitosus and to lectotypify the specimen in UPS. Korean Myelophycus specimens reported from Cheju Island and Wando were considered to belong to M. caespitosus based on the reported rbcL sequences.     

Origins of recently re‐established and newly discovered populations of the endangered butterfly Shijimiaeoides divinus (Lepidoptera: Lycaenidae) in Oita Prefecture,Japan

The endangered butterfly Shijimiaeoides divinus was believed to have been extirpated from Oita Prefecture, Kyushu, Japan, but was rediscovered in Taketa in recent years. This population is considered to have re‐established as a result of natural dispersal from Kumamoto, a neighboring prefecture located to the west of Oita. Furthermore, another population was recently found in Yufu, Oita Prefecture, which is an area where the species had never been recorded. To elucidate the origins of these two populations newly found from Oita Prefecture, their DNA sequences of the mitochondrial cytochrome c oxidase subunit I (COI) gene were compared with those of other S. divinus populations from Kumamoto Prefecture, Honshu and Korea. The results supported the hypothesis that the Taketa population originated from Kumamoto Prefecture. However, it was not clear whether this population originated from the natural dispersal or deliberate release of individuals. It was also found that the Yufu population was not established by the deliberate release of individuals from Honshu or Korea; however, it remained unclear whether the population of S. divinus was native to Yufu, or originated from other localities in Kyushu.     

A new timeframe for the diversification of Japan’s mammals

Aim The main Japanese islands are land‐bridge islands divided by the biogeographic division Blakiston’s Line and represent two natural laboratories for studying land‐bridge diversification. Colonization of the current mammal fauna has been dated to the middle to late Pleistocene using fossil evidence. The purpose of this paper is to apply a molecular clock to the genetic divergences between Japanese mammalian taxa and their sister mainland taxa to test the late Pleistocene land‐bridge colonization hypothesis. Location The main Japanese islands (Kyushu, Shikoku, Honshu and Hokkaido). Methods I used mitochondrial DNA (cytochrome b) and a species tree approach to estimate the divergence times of 24 Japanese non‐volant terrestrial mammal taxa and their mainland sister taxa using the program *beast . I then tested for evidence of non‐simultaneous divergence among these taxon‐pairs by controlling for expected coalescent stochasticity using the program Ms Bayes . Results Divergence events between taxa on Japan and their mainland sister taxa were significantly older than expected under the current paradigm, which is based on fossil data. Consistent with the land‐bridge colonization hypothesis, there was evidence of multiple divergence events. Main conclusions These results implicate a colonization timeframe that is older than posited by the current paradigm based on fossil evidence. However, these results are still consistent with the land‐bridge colonization hypothesis. Multiple periods of land‐bridge connectivity may account for the current mammalian fauna in Japan. In addition, half of the divergence time estimates in the Honshu–Shikoku–Kyushu region were clumped around 2.4 Ma, which might suggest a dramatic interchange period, concordant with a period of significant global cooling, when the first land bridge may have connected Japan to the mainland.     

Intraguild interactions and aphid predators: biological efficiency of Harmonia axyridis and Episyrphus balteatus

Introductions of the harlequin ladybird Harmonia axyridis into Belgium for aphid biological control have been followed by declines in native aphid natural enemies. We first examined, in laboratory and field conditions, the impact of larval mobility of either H. axyridis or Episyrphus balteatus, the most abundant native hoverfly in central Europe, on aphid suppression. The hoverfly larvae consumed more aphids (Megoura viciae) in a short term, whereas the ladybird ones were more efficient in a long term. We second investigated the intraguild interactions between H. axyridis and E. balteatus larvae and adults. In larva pairings between the two species in laboratory microcosms containing aphid (M. viciae) infested broad bean (Vicia faba) plants, H. axyridis had an intraguild predation (IGP) advantage over the hoverfly. When conspecific larvae were paired together on aphid‐infested plants, no cannibalism between them was detected. The presence of either H. axyridis or E. balteatus larvae on aphid‐infested plants negatively influenced the ovipositional behaviour of H. axyridis and E. balteatus females; lower numbers of laid eggs were recorded compared to control treatment. Moreover, eggs laid by E. balteatus females were also dropped as victims of predation mainly by H. axyridis larvae. Our results suggest that while the exotic ladybird was more efficient in aphid biological control, larvae and eggs of the native hoverfly species face increased IGP by H. axyridis, which would contribute, as a consequence, to the decline in E. balteatus population following invasion.     

Screening for resistance in apple cultivars to lightbrown apple moth, Epiphyas postvittana, and greenheaded leafroller, Planotortrix octo, and its relationship to field damage

The mature leaves of 38 apple (Malus domestica Borkh.) (Rosaceae) cultivars were screened for resistance to laboratory colonies of Epiphyas postvittana (Walker) and Planotortrix octo Dugdale (Lepidoptera: Tortricidae) by measuring larval survival and development rate, and pupal weight, in no‐choice laboratory bioassays in early summer. There were few cultivar effects on larval mortality but Nevis 1, A40R04T119, and ‘Sir Prize’ reduced survival of E. postvittana. Effects on development time and weight were correlated, and were integrated into a single measure of resistance R_c. R_c varied greatly between cultivars for both leafroller species, and identified 15 and 11 cultivars with partial resistance to E. postvittana and P. octo, respectively. A further experiment with three of the cultivars in midsummer, using the laboratory colonies in comparison with new colonies (from field collected larvae), produced similar results for E. postvittana, but there was no survival of new colony P. octo larvae on two cultivars. These results for P. octo are consistent with other recent research showing extreme cultivar resistance and critical colony and seasonal influences. A series of 4‐year field trials with the 38 cultivars showed poor correlation between laboratory and field resistance for the dominant leafroller species in the field, E. postvittana. The combined data, however, identified ‘Red Dougherty’, ‘Sir Prize’, and A40R04T119 as potentially useful in breeding for resistance to this species. The implications of these findings for integrated pest management (IPM) programmes and for the breeding of leafroller‐resistant apple cultivars are discussed.     

Japan's nationwide long‐term monitoring survey of seaweed communities known as the “Monitoring Sites 1000”: Ten‐year overview and future perspectives

“Monitoring Sites 1000” – Japan's long‐term monitoring survey was established in 2003, based on the Japanese Government policy for the conservation of biodiversity. Ecological surveys have been conducted on various types of ecosystems at approximately 1000 sites in Japan for 15 years now and are planned to be carried out for 100 years. Since 2008, seaweed communities had been monitored at six sites, featuring the kelp (e.g. Saccharina and Ecklonia; Laminariales) and Sargassum (Fucales) communities in the subarctic and temperate regions of Japan. Annual surveys were carried out during the season when these canopy‐forming seaweeds are most abundant. A non‐destructive quadrat sampling method, with permanent quadrats placed along transects perpendicular to the shoreline, was used to determine species composition, coverage, and vertical distribution of seaweeds at these sites; while destructive sampling was done every 5 years to determine biomass. The occurrence of canopy‐forming species Saccharina japonica (var. japonica) and Ecklonia cava have appeared to be stable at the Muroran (southwestern part of Hokkaido Island) and Shimoda (Pacific coast of middle Honshu Island) sites, respectively; whereas the coverage of Ecklonia radicosa (= Eckloniopsis radicosa) at the Satsuma‐Nagashima site in southern part of Kyushu Island was highly variable until its sudden disappearance from the habitat in 2016. Thalli of E. radicosa lost most of their blades through browsing by herbivorous fish, and thus, this may be one of the causes of the decline. A shift in the community structure related to environmental changes had also been observed at some other sites. Pre‐ and post‐disaster data revealed the impact of the 2011 earthquake and tsunami disasters, including a shift in the vertical distribution of Ecklonia bicyclis (= Eisenia bicyclis) to shallower depths at the Shizugawa site in the Pacific coast of northern Honshu Island, due to seafloor subsidence.     

Notes on the genus <Emphasis Type="Italic">Inocybe</Emphasis> of Japan. IV. Species having metuloids collected from Hokkaido,Honshu, and Kyushu

This fourth paper in the series considers five species of Inocybe occurring in Hokkaido, Honshu, and Kyushu. (1) Inocybe furfurea (section Tardae) is recorded from Hokkaido as new to Japan. (2) Inocybe luteola sp. nov. (section Tardae) is described from eastern Honshu (Chiba). It has smooth basidiospores and thick-walled caulocystidia descending to the middle of the stipe. (3) Inocybe napiformis sp. nov. [section Inocybe (= Cortinatae)], known from Hokkaido, appears close to Inocybe napipes, but characters of the metuloids distinguish the two species. (4) Inocybe grammata (section Marginatae) is recorded from Hokkaido and Nagano as new to Japan. (5) Inocybe pyriformis sp. nov. (section Marginatae) is described from Kyushu (Miyazaki). It has nodulose basidiospores and caulocystidia wholly covering the stipe surface. However, I. pyriformis is not typical in the section, lacking a marginate bulbous base in its stipe.     

Taxonomic review of Agraphydrus from Japan (Coleoptera: Hydrophilidae: Acidocerinae)

The Japanese species of the genus Agraphydrus are revised. Five species are recognized, including Agraphydrus ogatai sp. nov. from Honshu, Shikoku and Kyushu. The new species can be distinguished by the coarse ground punctures on the pronotum and elytra, and by the shape of parameres. All species except A. luteilateralis are redescribed. All species are diagnosed, relevant morphological characters are illustrated, distributional records are summarized and an updated key is provided.     

Observations of parasitoid behaviour in both no‐choice and choice tests are consistent with proposed ecological host range

The solitary larval endoparasitoid Eadya daenerys Ridenbaugh (Hymenoptera: Braconidae) is a proposed biocontrol agent of Paropsis charybdis Stål (Coleoptera: Chrysomelidae, Chrysomelinae), a pest of eucalypts in New Zealand. Eadya daenerys oviposition behaviour was examined in two assay types during host range testing, with the aim of improving ecological host range prediction. No‐choice sequential and two‐choice behavioural observations were undertaken against nine closely related species of New Zealand non‐target beetle larvae, including a native beetle, introduced weed biocontrol agents, and invasive paropsine beetles. No behavioural measure was significantly different between no‐choice and two‐choice tests. In sequential no‐choice assays the order of first presentation (target–non‐target) had no significant effect on the median number of attacks or the attack rate while on the plant. Beetle species was the most important factor. Parasitoids expressed significantly lower on‐plant attack rates against non‐targets compared to target P. charybdis larvae. The median number of attacks was always higher towards target larvae than towards non‐target larvae, except for the phylogenetically closest related non‐target Trachymela sloanei (Blackburn) (Coleoptera: Chrysomelidae, Chrysomelinae). Most non‐target larvae were disregarded upon contact, which suggests that the infrequent attack behaviour observed by two individual E. daenerys against Allocharis nr. tarsalis larvae in two‐choice tests and the frass of Chrysolina abchasica (Weise) was probably abnormal host selection behaviour. Results indicate that E. daenerys is unlikely to attack non‐target species apart from Eucalyptus‐feeding invasive paropsines (Chrysomelinae). Non‐lethal negative impacts upon less preferred non‐target larvae are possible if E. daenerys does attack them in the field; however, this is likely to be rare.     

Colonization history of the Japanese water shrewChimarrogale platycephala, in the Japanese Islands

To evaluate the intraspecific evolutionary history and local differentiation of the Japanese water shrewChimarrogale platycephala (Temminck, 1842), we an a lyzed the mitochondrial cytochromeb (Cytb) sequence divergence for samples from 55 localities in the Japanese is lands of Honshu and Kyushu. According to phylogenetic trees based on theCytb data, there were fourCytb haplotype lineages, which showed rough affinities with geographic areas, namely, Eastern/Central Honshu, the Kinki District of Western Honshu, the Chugoku District of Western Honshu, and Kyushu. However, in the alpine areas of the boundary between the Kinki and Chugoku Districts, complicated distribution patterns of theCytb haplotypes were revealed. Considering the present data and geological history in the Quaternary, we hypothesized the following evolutionary scenario. First, differ entiation and division into four primary ancestral geographic colonies of the shrews occurred in hypothetical refugia in the mid — late Pleistocene. Subsequently, rapid expansion occurred and caused the complicated distribution patterns of theCytb haplotypes in the boundary areas owing to the complex topography during the late stage of the Quaternary.