Identification of elmid larvae (Coleoptera: Elmidae) from Sanin District of Honshu,Japan, based on mitochondrial DNA sequences

Larvae of Elmidae from the Sanin District, Honshu, Japan, were classified into 14 types based on morphological features, of which 11 types were unidentified for species. Species or genus of the unidentified types were determined by comparing their mitochondrial cytochrome oxidase subunit I gene sequences with those of identified adult specimens. A new key to species/genera of elmid larvae was proposed.     

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.     

基于线粒体细胞色素c氧化酶亚基I基因序列的帘蛤科贝类分子系统发育研究

对21种帘蛤科贝类线粒体细胞色素c氧化酶亚基Ⅰ(cytochrome c oxidase subunit I,COI)基因核苷酸序列进行了分析,以探讨这一序列在种质鉴定、分子系统发生研究中的应用价值。测序结果表明,所有物种扩增片段长度均为707 bp(含引物),序列A+T含量(62.4%—67.8%)明显高于G+C含量。物种间共有变异位点379个,其中简约信息位点334个;此区段共编码235个氨基酸,种间共有氨基酸变异位点100个。以COI基因片段序列为标记,用中国蛤蜊(Mactra chinensis)作外群,构建了35种帘蛤科贝类(其中14种贝类COI序列从GenBank下载)的系统发生树,结合拓扑结构分析和序列比对分析,结果表明:支持将短文蛤(Meretrix petechinalis)和丽文蛤(M.lusoria)订为文蛤(M.meretrix)的同物异名的观点,建议将丽文蛤和短文蛤订为文蛤的地理亚种;支持将薄片镜蛤(Dosinia corrugata)和D.angulosa订为2个独立种的观点;认为将波纹巴非蛤(Paphia undulata)和织锦巴非蛤(P.textile)订为2个独立种是合适的。COI基因序列含有丰富的遗传信息,适合作为帘蛤科贝类种群遗传结构和系统发生研究的分子标记。     

Different phylogeographic patterns in two Japanese Silpha species (Coleoptera: Silphidae) affected by climatic gradients and topography

To reveal differences in phylogeographic patterns of flightless insect species occurring in different regions of Japan, we studied the phylogeography and demographic history of Silpha beetles occurring in cool-temperate habitats of two major islands, Honshu and Hokkaido, using sequences of the mitochondrial cytochrome oxidase subunit I (COI) gene. Honshu has a more mountainous topography, and cool-temperate habitats occur discontinuously, whereas Hokkaido, located to the north of Honshu, has more continuous cool-temperate habitats. A species endemic to Honshu, S. longicornis occurs on Honshu, whereas S. perforata occurs on Hokkaido and the East Asian continent. Our results indicate that the ancestors of S. longicornis colonized Honshu via a south-west route c . 0.7 Mya and the species has highly divergent populations in isolated mountainous areas of Honshu, whereas S. perforata colonized Hokkaido via a northern route less than 90 000 years ago and has less divergent geographic populations. During the last glacial period, S. perforata was probably restricted to refugia in southern Hokkaido and later expanded into northern Hokkaido, whereas S. longicornis populations existed in many isolated refugia, probably because of the complex topography of Honshu. Thus, our study demonstrates that, even between closely related species, interactions among biology, latitudinal climatic gradients and topography can produce different phylogeographic patterns.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 98 , 452–467.     

Discovery of swimming larvae in Elmidae (Coleoptera: Byrrhoidea)

The aquatic larvae of the family Elmidae (Coleoptera) have been considered unable to swim because they lack swimming setae on their body and legs. We discovered that riffle beetle larvae (Leptelmis gracilis Sharp) are in fact able to swim. After opening the tuft‐like gills on the last abdominal segment they swim by repeatedly bending the abdomen into a U‐shape very quickly and in a wavy manner, while their head and thorax move anteriorly, resulting in a sigmoidal body shape. The flat bodies of L. gracilis larvae could be advantageous for swimming compared to the cylindrical or semicylindrical bodies of larvae in related genera.     

Molecular systematics of the Chrysoperla carnea group (Neuroptera: Chrysopidae) in Europe

The green lacewing Chrysoperla carnea is a complex of cryptic species whose differentiation has been so far based upon morphology, ecophysiology, behaviour and preliminary mitochondrial DNA data using cytochrome oxidase subunit II (COII) and NADH dehydrogenase subunit II. In this work we extended the DNA data by screening nucleotide sequences of COII, cytochrome oxidase I, cytochrome b and the large ribosomal subunit of the mtDNA. These new data suggest that C. carnea s.s. is a well-supported, separate taxon, but that other taxa of the complex are not consistently differentiated by the current DNA data.     

DNA-based identification of Alaska skates (Amblyraja, Bathyraja and Raja: Rajidae) using cytochrome c oxidase subunit I (coI) variation

Variation at the mitochondrial cytochrome c oxidase subunit I (mt-COI) gene was examined in 15 species of North Pacific skates. Thirteen species had unique sequences, indicating that a DNA-based barcoding approach may be useful for species identification.     

Polymorphism observed in mitochondrial genes of red flour beetle,Tribolium castaneum (Coleoptera: Tenebrionidae) of different origin in laboratory cultures

Tribolium castaneum, a pest of stored grain, has a uniform morphology, preventing the visual identification of strains from different areas. Polymorphisms in the nucleotide sequences of the mitochondrial genes of this species were examined, and combined into seven haplotypes among the test insect specimens originating from Japan, Thailand, and Canada. These results suggested the potential for geographical differentiation.     

Physical gills in Elmidae (Coleoptera: Byrrhoidea): Structure and evolutionary pattern of plastron in Stenelmis and related genera

Many species within Elmidae (Coleoptera: Byrrhoidea) have plastrons composed of flattened setae. However, some genera display fine plastrons on the epicuticle, called plastron hairs. In Japanese elmids, members of the genera Stenelmis, Ordobrevia, Nomuraelmis and Leptelmis bear ventral plastron hairs. Based on a maximum likelihood tree including most Japanese genera within Elmidae, we found that these genera are monophyletic and that plastron hairs are a derived character in Elmidae. We also found that the genus Graphelmis bears jigsaw puzzle‐like plastron scales with plastron hair‐like projections, and is sister to the group with plastron hairs.     

Genetic diversity of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) populations and presence of the B biotype and a non-B biotype that can induce silverleaf symptoms in squash, in Uganda

The extent of genetic variability and host‐plant distribution of Bemisia tabaci (Gennadius) genotypes colonising cultivated and uncultivated plant species occurring adjacent to cassava fields in selected cassava‐producing areas of Uganda in 2003/04 were investigated using the mitochondrial cytochrome oxidase I (mtCOI) gene as the molecular marker. Eight genotype clusters, Ug1–Ug8, which are supported by high bootstrap values (≥80), at 3–18% nt divergence, were revealed among the collective Ugandan B. tabaci populations. Ug1 and Ug2 (both cassava‐associated) and Ug8 (sweetpotato‐associated) have been reported previously in Uganda. Ug3 was genetically dissimilar to B. tabaci described elsewhere and colonised a single species, Ocimum gratissimum. Ug4–Ug7 formed four closely related subclusters (93–97% nt identity) and diverged by 15–18% from Ug1, Ug2, Ug3 and Ug8, respectively. Ug4 had as its closest relatives (at 97–99% nt identity) the Ivory Coast okra biotype, whereas genotypes Ug5 and Ug6 had as their closest relatives (at 95–99% and 99% nt identity, respectively) the Mediterranean–North Africa–Middle East (MED‐NAFR‐ME) biotypes, which also include the well‐studied B and Q biotypes. Ug7 was closely related (at 98–99% nt identity) to biotype Ms from the Reunion Island in the Indian Ocean. Ug4 colonised Cucurbita pepo, Cucurbita sativus, Leonotis nepetifolia and Pavonia urens, while Ug7 colonised Commelina benghalensis, Gossypium hirsutum and Phaseolus vulgaris. Ug6, the B‐biotype‐like genotype colonised Abelmoschus esculentus and C. benghalensis only. None of Ug4–Ug7 genotypes was found associated with, or colonising, cassava or sweetpotato plants. In addition to colonising sweetpotato, the Ug8 genotypes colonised Lycopersicon esculentum and L. nepetifolia. Ug6 and Ug7, both members of the B biotype/B‐like cluster, induced silverleaf symptoms on Cucurbita sp. The discovery of five previously identified B. tabaci genotype clusters, Ug3–Ug7, in Uganda, among which are some of the world's most economically important biotypes, namely B and Q, is particularly significant in the spread of geminiviruses with devastating effects to crop production in Africa.     

Data on molecular taxonomy and genetic diversification of the European Hermit beetles, a species complex of endangered insects (Coleoptera: Scarabaeidae, Cetoniinae, Osmoderma)

A molecular analysis was carried out on the European hermit beetles (the Osmoderma eremita species complex) to explore their genetic diversification and the robustness of previous morphologically based taxonomic arrangements. Complete sequences of mtDNA cytochrome C oxidase I gene were obtained from 26 individuals. Mean levels of interspecific sequence divergence ranged from 0.044 to 0.186. The results indicate a clear-cut distinction between two clades. The first one includes the W-European O. eremita Scopoli 1763 ; and the two Italian endemic taxa Osmoderma italicum Sparacio, 2000 and Osmoderma cristinae Sparacio, 1994 ; from southern peninsular Italy and Sicily, respectively. The second one includes the widespread E-European Osmoderma barnabita ; and the southern Balcanic Osmoderma lassallei Baraud and Tauzin, 1991 from Greece and European Turkey. Within the two clades, molecular and morphological data well support a specific rank for O. lassallei and O. barnabita on one side, and for O. eremita and O. cristinae on the other side, while the taxonomic position of O. italicum , more closely related to O. eremita , is still uncertain. Current geographical distribution, interspecific genetic diversification, and very low levels of intraspecific genetic divergence in western European populations of O. eremita sensu stricto are hypothesized to be the result of multiple speciation events (mainly occurred in refugial forest areas of the Italian and Balkan peninsulas and Sicily before and during the Pleistocene glacial peaks), followed by fast post-glacial northward and westward expansion of some species.     

Phylogeography of Simulium siamense Takaoka and Suzuki complex (Diptera: Simuliidae) in Thailand

The phylogeography of Simulium siamense complex was inferred from mitochondrial DNA sequences. A 586‐bp fragment of the cytochrome oxidase I was sequenced for 92 individuals from 13 populations throughout Thailand, representing five cytoforms (A, B, C, F and G). The cytoforms are not genetically different at the molecular level except for cytoform B, which is genetically distinct from the others. This might indicate that cytoform B is a distinct species. Further morphological and molecular work using other genes is needed to clarify this. Our results also argue for the need of integrated approach, both cytological and molecular studies to understanding biodiversity of black flies. The star‐like shape of the mtDNA genealogy is consistent with the sudden population expansion of the mismatch distribution analysis and large negative values of Fu's F_s and Tajima's D‐tests, indicating a population demographic expansion. The expansion time is estimated to be in the late Pleistocene (about 120 000 years ago). Therefore, the overall low level of genetic structure could be due to sharing a recent history. The ancestral haplotype was found in the mountainous area in northeastern Thailand, suggesting that this area could have been the refugium of the species complex during the Pleistocene glaciations. Our results are consistent with previous findings about population expansion in response to the Pleistocene climatic change, thus revealing the importance of this historical event in shaping the genetic structure and diversity of Southeast Asian mainland species.     

Barcoding,molecular taxonomy,and exploration of the diversity of shrews (Soricomorpha: Soricidae) on Mount Nimba (Guinea)

We tested the efficiency of cytochrome oxidase I (COI)‐barcoding as a taxonomic tool to discriminate and identify sympatric shrew species on Mount Nimba (Guinea). We identified 148 specimens at the species level using morphological characters and comparison with type specimens, including several taxa from Mount Nimba. We identified ten morphospecies and tested aspects of genetic diversity and monophyly using genetic data from three mitochondrial (16S, cytochrome b, and COI) and one nuclear marker (the breast cancer gene, BRCA). Nine morphospecies were validated under the phylogenetic and genetic species concepts, including the recently diverged species Crocidura buettikoferi, Crocidura theresae, and Crocidura grandiceps. Under the same concepts, our analyses revealed the presence of two cryptic species amongst animals identified as Crocidura muricauda. We then tested the efficiency of barcoding thanks to commonly used phenetic methods, with the 148 specimens representing 11 potentially valid species based on morphological and molecular data. We show that COI‐barcoding is a powerful tool for shrew identification and can be used for taxonomic surveys. The comparison of genetic divergence values shows the presence of a barcoding gap (i.e. difference between the highest intraspecific and the lowest interspecific genetic divergence values). Given that only a few COI sequences are available for Afrotropical shrews, our work is an important step forward toward their enrichment. We also tested the efficiency of the three other sequenced markers and found that cytochrome b is as efficient as COI for barcoding shrews. © 2012 The Linnean Society of London, Zoological Journal of the Linnean Society, 2012, 166 , 672–687.     

PCR gut analysis reveals that Tenuiphantes tenuis (Araneae: Linyphiidae) is a potentially significant predator of Argentine stem weevil,Listronotus bonariensis (Coleoptera: Curculionidae), in New Zealand pastures

Polymerase chain reaction (PCR) gut analysis was conducted on specimens of the introduced spider Tenuiphantes tenuis collected from dairy pasture in Canterbury, New Zealand. PCR primers were specifically designed to amplify a fragment of the mitochondrial gene cytochrome c oxidase subunit 1 (COI) from Listronotus bonariensis and revealed that this major pasture pest species is consumed in the field by T. tenuis. The field predation rate of L. bonariensis by T. tenuis was estimated from our PCR results together with published data on the degradation of DNA and the density of T. tenuis in Canterbury pastures. We found that T. tenuis is a potentially significant predator of L. bonariensis in New Zealand pastures.     

Colonization, dispersal, and hybridization influence phylogeography of North Atlantic sea urchins (Strongylocentrotus droebachiensis)

We used frequency-based and coalescent-based phylogeographic analysis of sea urchin (Strongylocentrotus droebachiensis) mitochondrial DNA (mtDNA) sequences and previously published microsatellite data to understand the relative influence of colonization and gene flow from older (north Pacific) and younger (northeast Atlantic) sea urchin populations on genetic variation in the northwest Atlantic. We found strong evidence of survival of northwestern Atlantic populations in local Pleistocene glacial refugia: most haplotypes were the same as or closely related to Pacific haplotypes, with deep gene genealogies that reflect divergence times within the northwestern Atlantic that are much older than the last glacial maximum. We detected gene flow across the North Atlantic in the form of haplotypes shared with or recently descended from European populations. We also found evidence of significant introgression of haplotypes from a closely related species (S. pallidus). The relative magnitude of gene flow estimated by coalescent methods (and the effective population size differences among oceanic regions) depended on the genetic marker used. In general, we found very small effective population size in the northeastern Atlantic and high trans-Arctic gene flow between the Pacific and northwestern Atlantic. Both analyses suggested significant back-migration to the Pacific. However, microsatellites more strongly reflected older Pacific migration (with similar effective population sizes across the Arctic), whereas mtDNA sequences appeared to be more sensitive to recent trans- Atlantic dispersal (with larger differences in effective population size). These differences across marker types might have several biological or methodological causes, and they suggest caution in interpretation of the results from a single locus or class of markers.     

Brazilian populations of Drosophila maculifrons (Diptera: Drosophilidae): low diversity levels and signals of a population expansion after the Last Glacial Maximum

The Quaternary period was marked by considerable changes in climate. Such palaeoclimatic changes affected the population dynamics of many species, both in the Northern and in the Southern Hemisphere. However, the extent of these impacts on the demographic patterns of Neotropical species presenting different ecological requirements remains unclear. Drosophila maculifrons DUDA 1947 belongs to the guaramunu group of Drosophila and represents a potential indicator of the genetic consequences caused by the climatic fluctuations of the Quaternary, because it seems to be sensitive to temperature and humidity shifts. The aim of this study was to evaluate the evolutionary processes subjacent to the patterns of intraspecific diversity and structure of different populations of D. maculifrons. In total, 152 individuals were collected in the south and south‐east Brazil. Phylogenetic and phylogeographical analyses were performed based on sequences of COI and COII mitochondrial genes. In general, the results pointed to Brazilian populations of D. maculifrons being extremely impoverished in terms of mitochondrial diversity and population structure, which could be explained by a recent population expansion event dated to approximately 12 000 years ago. In fact, with the assistance of species palaeo‐distribution modelling strategies, it was possible to infer that most of the sampled region did not present the D. maculifrons environmental suitability requirements at least during the period of the Last Glacial Maximum. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 112 , 55–66.     

Resurrection of Rabdophaga salicivora Shinji (Diptera: Cecidomyiidae), a Japanese gall midge formerly misidentified as a North American species, Rabdophaga rigidae (Osten Sacken), with observations on the phylogenetic relationships of its populations in Japan and the Russian Far East

Intercontinental biotic connections between Eurasia and North America are common in many gall midge genera (Diptera: Cecidomyiidae), but only a few species have been recorded from both continents. In Japan, four gall midge species had been previously considered to be identical to North American species, but three of these cases have already been disproved. We examined the remaining species, Rabdophaga rigidae, which had been originally described from Japan as Rabdophaga salicivora in 1938, later recorded from the Russian Far East in 1967, and synonymized with a North American species, R. rigidae, in 1982. Morphological features and partial sequence data of the mtDNA cytochrome oxidase subunit I (COI) region suggested that the Japanese species is a distinct species and is identical to the species recorded from the Russian Far East. We therefore apply the original name, R. salicivora, to the Japanese and the Russian species. In addition, on the basis of a molecular phylogenetic analysis, we conclude that R. salicivora possibly came to the Japanese Archipelago through the Korean Peninsula and established itself first in the southern parts of Japan. Then, it expanded its distribution range to northern parts of Honshu, but could not reach Hokkaido, probably because of the Tsugaru Strait between Honshu and Hokkaido.     

Revision of the lygaeid genus Nysius (Heteroptera: Lygaeidae: Orsillinae) of Japan,with description of a new species

Japanese species of the lygaeid–orsilline genus Nysius were revised, and five species including a new species, N. hidakai, were recognized. Male and female genitalia of all the species were illustrated, and a key to species of Japanese Nysius was provided. The DNA barcoding information of each species except N. expressus were recorded.     

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.