DNA Barcoding of Japanese Click Beetles (Coleoptera,Elateridae)

Click beetles (Coleoptera: Elateridae) represent one of the largest groups of beetle insects. Some click beetles in larval form, known as wireworms, are destructive agricultural pests. Morphological identification of click beetles is generally difficult and requires taxonomic expertise. This study reports on the DNA barcoding of Japanese click beetles to enable their rapid and accurate identification. We collected and assembled 762 cytochrome oxidase subunit I barcode sequences from 275 species, which cover approximately 75% of the common species found on the Japanese main island, Honshu. This barcode library also contains 20 out of the 21 potential pest species recorded in Japan. Our analysis shows that most morphologically identified species form distinct phylogenetic clusters separated from each other by large molecular distances. This supports the general usefulness of the DNA barcoding approach for quick and reliable identification of Japanese elaterid species for environmental impact assessment, agricultural pest control, and biodiversity analysis. On the other hand, the taxonomic boundary in dozens of species did not agree with the boundary of barcode index numbers (a criterion for sequence-based species delimitation). These findings urge taxonomic reinvestigation of these mismatched taxa.     

Two New Species of the Genus Melanotus Eschscholtz (Coleoptera: Elateridae) from Korea

Two new species, M. insularis sp. nov. and M. woonhahi sp. nov., of the genus Melanotus Eschscholtz from Korea are described. The external characteristics of both species are similar to a Japanese species, M. senilis Candeze. In addition to the difference of male genitalia, however, M. insularis can be distinguished by the strong body punctures from M. woonhahi and by short antennae and small and sparse body puncture from M. senilis.     

五种槽缝叩甲超微形态结构的比较研究 (鞘翅目:叩甲科)

通过扫描电镜观察了槽缝叩甲属Agrypnus 5种的触角、前胸背板、中胸小盾片、鞘翅、前胸腹板、前胸侧板、中胸腹窝、跗节等部位的超微形态结构,提出了5种槽缝叩甲超微形态的区别特征,发现了槽缝叩甲体表鳞片毛上的纵脊数目和走向、中胸腹窝的结构与形状、体表刻点的形态等具有明显的种间差异,具有重要的分类学意义。讨论了硬结构昆虫扫描样品制备的程序,叩甲的叩头关节、触角槽、跗节槽的形成与功能,以及超微形态结构在叩甲分类上的价值等。     

Phylogeny of the Thylacosterninae (Coleoptera, Elateridae)

The phylogeny of the Elateridae subfamily Thylacosterninae was studied using morphological characters and mitochondrial 16S rDNA sequences. Five monophyletic groups were recovered: the Asian Cussolenis , the American Pterotarsus , Balgus and Thylacosternus , and the African " Cussolenis ", which is described here as Lumumbaia new genus Muona & Vahtera (type-species Cussolenis attenuatus Fleutiaux, 1925). The following new combinations are made: Lumumbaia africanus (Fleutiaux, 1897) [ Soleniscus ], Lumumbaia praeustus (Fleutiaux, 1926) [ Cussolenis ], Lumumbaia nigripes (Fleutiaux, 1926) [ Cussolenis ], Lumumbaia robustus (Fleutiaux, 1925) [ Cussolenis ], Lumumbaia attenuatus (Fleutiaux, 1925) [ Cussolenis ] and Lumumbaia notabilis (Fleutiaux, 1926) [ Cussolenis ]. A key to the genera of the subfamily is provided. The data were efficient in recovering generic limits within the group. At species level the mitochondrial 16S rDNA data seemed to work unpredictably, either agreeing or not with traditional species-level limits based on male genitalia and body structure. The evolution of bioluminescence is optimized as a feature originating in the ancestors of clicking elateroids.  © The Willi Hennig Society 2009.     

Phylogenetic relationships of click beetles (Coleoptera: Elateridae) inferred from 28S ribosomal DNA: insights into the evolution of bioluminescence in Elateridae

Although the taxonomy of click beetles (family Elateridae) has been studied extensively, inconsistencies remain. We examine here the relationships between species of Elateridae based on partial sequences of nuclear 28S ribosomal DNA. Specimens were collected primarily from Japan, while luminous click beetles were also sampled from Central and South America to investigate the origins of bioluminescence in Elateridae. Neighbor-joining, maximum-parsimony, and maximum-likelihood analyses produced a consistent basal topology with high statistical support that is partially congruent with the results of previous investigations based on the morphological characteristics of larvae and adults. The most parsimonious reconstruction of the "luminous" and "nonluminous" states, based on the present molecular phylogeny, indicates that the ancestral state of Elateridae was nonluminous. This suggests that the bioluminescence in click beetle evolved independent of that of other luminous beetles, such as Lampyridae, despite their common mechanisms of bioluminescence.     

Analyzing Mosquito (Diptera: Culicidae) Diversity in Pakistan by DNA Barcoding

Background

Although they are important disease vectors mosquito biodiversity in Pakistan is poorly known. Recent epidemics of dengue fever have revealed the need for more detailed understanding of the diversity and distributions of mosquito species in this region. DNA barcoding improves the accuracy of mosquito inventories because morphological differences between many species are subtle, leading to misidentifications.

Methodology/Principal Findings

Sequence variation in the barcode region of the mitochondrial COI gene was used to identify mosquito species, reveal genetic diversity, and map the distribution of the dengue-vector species in Pakistan. Analysis of 1684 mosquitoes from 491 sites in Punjab and Khyber Pakhtunkhwa during 2010–2013 revealed 32 species with the assemblage dominated by Culex quinquefasciatus (61% of the collection). The genus Aedes (Stegomyia) comprised 15% of the specimens, and was represented by six taxa with the two dengue vector species, Ae. albopictus and Ae. aegypti, dominant and broadly distributed. Anopheles made up another 6% of the catch with An. subpictus dominating. Barcode sequence divergence in conspecific specimens ranged from 0–2.4%, while congeneric species showed from 2.3–17.8% divergence. A global haplotype analysis of disease-vectors showed the presence of multiple haplotypes, although a single haplotype of each dengue-vector species was dominant in most countries. Geographic distribution of Ae. aegypti and Ae. albopictus showed the later species was dominant and found in both rural and urban environments.

Conclusions

As the first DNA-based analysis of mosquitoes in Pakistan, this study has begun the construction of a barcode reference library for the mosquitoes of this region. Levels of genetic diversity varied among species. Because of its capacity to differentiate species, even those with subtle morphological differences, DNA barcoding aids accurate tracking of vector populations.     

DNA Barcoding and Species Boundary Delimitation of Selected Species of Chinese Acridoidea (Orthoptera: Caelifera)

We tested the performance of DNA barcoding in Acridoidea and attempted to solve species boundary delimitation problems in selected groups using COI barcodes. Three analysis methods were applied to reconstruct the phylogeny. K2P distances were used to assess the overlap range between intraspecific variation and interspecific divergence. “Best match (BM)”, “best close match (BCM)”, “all species barcodes (ASB)” and “back-propagation neural networks (BP-based method)” were utilized to test the success rate of species identification. Phylogenetic species concept and network analysis were employed to delimitate the species boundary in eight selected species groups. The results demonstrated that the COI barcode region performed better in phylogenetic reconstruction at genus and species levels than at higher-levels, but showed a little improvement in resolving the higher-level relationships when the third base data or both first and third base data were excluded. Most overlaps and incorrect identifications may be due to imperfect taxonomy, indicating the critical role of taxonomic revision in DNA barcoding study. Species boundary delimitation confirmed the presence of oversplitting in six species groups and suggested that each group should be treated as a single species.     

DNA Barcoding in Pencilfishes (Lebiasinidae: Nannostomus) Reveals Cryptic Diversity across the Brazilian Amazon

Nannostomus is comprised of 20 species. Popularly known as pencilfishes the vast majority of these species lives in the flooded forests of the Amazon basin and are popular in the ornamental trade. Among the lebiasinids, it is the only genus to have undergone more than one taxonomic revision. Even so, it still possesses poorly defined species. Here, we report the results of an application of DNA barcoding to the identification of pencilfishes and highlight the deeply divergent clades within four nominal species. We surveyed the sequence variation in the mtDNA cytochrome c oxidase subunit I gene among 110 individuals representing 14 nominal species that were collected from several rivers along the Amazon basin. The mean Kimura-2-parameter distances within species and genus were 2% and 19,0%, respectively. The deep lineage divergences detected in N. digrammus, N. trifasciatus, N. unifasciatus and N. eques suggest the existence of hidden diversity in Nannostomus species. For N. digrammus and N. trifasciatus, in particular, the estimated divergences in some lineages were so high that doubt about their conspecific status is raised.     

Exploring the Leaf Beetle Fauna (Coleoptera: Chrysomelidae) of an Ecuadorian Mountain Forest Using DNA Barcoding

Background

Tropical mountain forests are hotspots of biodiversity hosting a huge but little known diversity of insects that is endangered by habitat destruction and climate change. Therefore, rapid assessment approaches of insect diversity are urgently needed to complement slower traditional taxonomic approaches. We empirically compare different DNA-based species delimitation approaches for a rapid biodiversity assessment of hyperdiverse leaf beetle assemblages along an elevational gradient in southern Ecuador and explore their effect on species richness estimates.

Methodology/Principal Findings

Based on a COI barcode data set of 674 leaf beetle specimens (Coleoptera: Chrysomelidae) of 266 morphospecies from three sample sites in the Podocarpus National Park, we employed statistical parsimony analysis, distance-based clustering, GMYC- and PTP-modelling to delimit species-like units and compared them to morphology-based (parataxonomic) species identifications. The four different approaches for DNA-based species delimitation revealed highly similar numbers of molecular operational taxonomic units (MOTUs) (n = 284–289). Estimated total species richness was considerably higher than the sampled amount, 414 for morphospecies (Chao2) and 469–481 for the different MOTU types. Assemblages at different elevational levels (1000 vs. 2000 m) had similar species numbers but a very distinct species composition for all delimitation methods. Most species were found only at one elevation while this turnover pattern was even more pronounced for DNA-based delimitation.

Conclusions/Significance

Given the high congruence of DNA-based delimitation results, probably due to the sampling structure, our study suggests that when applied to species communities on a regionally limited level with high amount of rare species (i.e. ~50% singletons), the choice of species delimitation method can be of minor relevance for assessing species numbers and turnover in tropical insect communities. Therefore, DNA-based species delimitation is confirmed as a valuable tool for evaluating biodiversity of hyperdiverse insect communities, especially when exact taxonomic identifications are missing.     

Antennal sensilla of the click beetle,Melanotus villosus (Geoffroy) (Coleoptera: Elateridae)

The typology, number and placement of antennal sensilla of the click beetle Melanotus villosus (Geoffroy) (Coleoptera: Elateridae) were studied using scanning electron microscopy. On both the males and females the antennae are made up of the scape, pedicel and nine flagellomeres. Two types of basiconic sensilla, three types of trichoid sensilla, one type of styloconic sensilla, one type of chetoid sensilla, dome-shaped sensilla, grooved pegs, and Böhm sensilla all appear on the antennae of the beetles of both sexes, with the exception of trichoid sensilla type II, whose large number (average of 1635 hairs per antenna) was found only in male beetles. Sensilla trichodea type II evidently respond to the sex pheromone produced by the female beetle. Unlike the other two click beetles, studied up till now, Agriotes obscurus and Limonius aeruginosus, the trichoid and basiconic sensilla of M. villosus, whose proven or assumed function is olfactory, are located predominantly on the flagellomeres ventral extensions. It is assumed that the placement of the olfactory sensilla, mainly on the ventral side of M. villosuss antennae, and their more or less even distribution on the flagellomeres, can be seen as morphological adaptation of this species of insect, whose specific behavioural reaction of olfactory searching is flying, both before and after contact with an odour plume.     

DNA Barcoding Reveals Cryptic Diversity within Commercially Exploited Indo-Malay Carangidae (Teleosteii: Perciformes)

Background

DNA barcodes, typically focusing on the cytochrome oxidase I gene (COI) in many animals, have been used widely as a species-identification tool. The ability of DNA barcoding to distinguish species from a range of taxa and to reveal cryptic species has been well documented. Despite the wealth of DNA barcode data for fish from many temperate regions, there are relatively few available from the Southeast Asian region. Here, we target the marine fish Family Carangidae, one of the most commercially-important families from the Indo-Malay Archipelago (IMA), to produce an initial reference DNA barcode library.

Methodology/Principal Findings

Here, a 652 bp region of COI was sequenced for 723 individuals from 36 putative species of Family Carangidae distributed within IMA waters. Within the newly-generated dataset, three described species exhibited conspecific divergences up to ten times greater (4.32–4.82%) than mean estimates (0.24–0.39%), indicating a discrepancy with assigned morphological taxonomic identification, and the existence of cryptic species. Variability of the mitochondrial DNA COI region was compared within and among species to evaluate the COI region''s suitability for species identification. The trend in range of mean K2P distances observed was generally in accordance with expectations based on taxonomic hierarchy: 0% to 4.82% between individuals within species, 0% to 16.4% between species within genera, and 8.64% to 25.39% between genera within families. The average Kimura 2-parameter (K2P) distance between individuals, between species within genera, and between genera within family were 0.37%, 10.53% and 16.56%, respectively. All described species formed monophyletic clusters in the Neighbour-joining phylogenetic tree, although three species representing complexes of six potential cryptic species were detected in Indo-Malay Carangidae; Atule mate, Selar crumenophthalmus and Seriolina nigrofasciata.

Conclusion/Significance

This study confirms that COI is an effective tool for species identification of Carangidae from the IMA. There were moderate levels of cryptic diversity among putative species within the central IMA. However, to explain the hypothesis of species richness in the IMA, it is necessary to sample the whole family across their broad geographic range. Such insights are helpful not only to document mechanisms driving diversification and recruitment in Carangidae, but also to provide a scientific framework for management strategies and conservation of commercially-important fisheries resources.     

Food habits of pasture wireworm,Conoderus exsul (Coleoptera: Elateridae)

Abstract

The food preferences and rates of growth of the omnivorous pasture wireworm, Conoderus exsul (Sharp) were studied. Larvae of soil-living insects were preferred to germinating maize seed, and wireworm growth rate was greater on insect compared with plant food. In glasshouse pot trials, wireworms increased their attack rate as the density of grass grub (Costelytra zealandica (White)) or white-fringed weevil (Graphthognathus leucoloma Boheman) larvae was increased. In field experiments that compared grass grub numbers in cages accessible to wireworms or protected from predators, predation varied with season from a low of 15% in late summer up to 82% in late autumn. White-fringed weevil larval mortality was increased by addition of wireworms in two field experiments, but not affected in a third trial where initial white-fringed weevil density was low. Grass grub mortality was increased by the presence of wireworms in field plots. Adult C. exsul were observed feeding on sweetcorn pollen.     

DNA barcoding identification and genetic diversity of bamboo shoot wireworms (Coleoptera: Elateridae) in South China

Wireworms are recognized as economically important oligophagous pests of Phyllostachys bamboo shoot, which have caused a huge economic loss in South China. Studies of bamboo shoot wireworm control and management were seriously hampered by the lack of reliable identification of larvae. DNA barcoding has been proved to be an important and useful tool in species identification of morphologically cryptic insects. Accurate knowledge of damage-causing species and the phylogenic structure of elaterids will provide insight into their sustainable and effective management. Here, we use interspecific variation in COI and EF1-a as a robust method of identification and consider the intra- and interspecific genetic variation of bamboo wireworms. This approach has revealed that three Melanotus larvae occurred in Phyllostachys forest. The dominance of one species in different sampling location suggests there maybe underlying difference in habit and host plant preference. These data provide a starting point for determining the distribution of wireworm species feeding on bamboo shoots in South China. The inclusions would further aid identification of wireworm species and relationships between certain wireworm species with specific environments.     

Wireworm (Coleoptera: Elateridae) identity, monitoring and damage in sugarcane

Abstract Wireworms (Coleoptera: Elateridae) damage the eyes and young shoots of germinating cane setts in many growing regions. Insecticides can be applied at planting to avoid damage. However, there are no guidelines for deciding on whether treatment is warranted and a sampling method to detect wireworms before planting has not been developed. Baits of rolled oats were tested as a method for detecting wireworms in preplant fallows. In 1995 at Bundaberg, the reduction in crop establishment when treatment was withheld was positively correlated with the catch of wireworms at baits before planting. However, no similar relationship was seen in 1996 or at Mackay in either 1995 or 1996: severe damage was recorded on one farm at Mackay where no wireworms had been found. A sampling plan was developed to estimate wireworm numbers at baits with a fixed level of precision. Wireworms collected at the baits and in samples from established canefields in Queensland and New South Wales included five named and 21 unnamed species from five genera, the most abundant being Agrypnus , Conoderus and Heteroderes . Possible barriers to the adoption of baiting as a decision-support tool are discussed. Development of a reliable monitoring system would require considerably more research, which could not be justified at present.     

On Findings of Three Western Palaearctic Species of the Weevil Subfamily Entiminae (Coleoptera,Curculionidae) in Yakutia

Entomological Review - Three common Western Palaearctic species of the weevil subfamily Entiminae are recorded from Yakutia for the first time: Otiorhynchus velutinus Germ., O. raucus (F.) and...     

New click beetles (Coleoptera: Elateridae) from the cretaceous of Russia and Kazakhstan

A new tribe, two new genera, and seven new species of click beetles are described: Pollostelaterini, trib. nov., Pollostelater baissensis, gen. et sp. nov. from the Lower Cretaceous of Transbaikalia (Baisa locality), five new species in the genus Cryptocoelus Dolin et Nel C. sinitshenkovae, sp. nov. (Romanovka locality), C. shcherbakovi, sp. nov., C. baissensis, sp. nov., C. lukashevichae, sp. nov., C. dolini, sp. nov. (all from Baisa locality), and Turonelater giganteus, gen. et sp. nov. from the Turonian of Southern Kazakhstan (Kzyl-Dzhar locality).     

Click Beetles (Coleoptera,Elateridae) in Agrolandscapes of Northwestern Russia

Entomological Review - A total of 44 species of click beetles (Coleoptera, Elateridae) were recorded in the agricultural landscapes of Northwestern Russia; 23 species were found in various crop...     

地黄属植物的DNA条形码研究

地黄属(Rehmannia)为玄参科(Scrophulariaceae)药用植物,广泛分布于中国中东部及北部地区。由于地黄属植物经历了快速成种,导致其属内物种间形态性状差异较小,运用传统的形态学分类方法已难以准确地鉴定物种,近年来迅速发展起来的DNA条形码技术为快速、准确地鉴别物种提供了新思路。本研究选用3个叶绿体DNA非编码区片段(trn L-trn F、trn M-trn V和trn S-trn G)及核基因ITS片段,运用PWG-distance和TreeBuilding两种方法对地黄属5个物种75个个体进行了DNA条形码分析。结果表明:单个叶绿体DNA片段或核基因ITS片段对地黄属物种的鉴别率较低(0%~20%),组合的叶绿体DNA片段分辨能力虽然高于单个DNA片段,但并不能将地黄属5个物种完全区分开;trn S-trn G+ITS片段组合的分辨率可达100%,能够将地黄属5个物种准确区分,与所有叶绿体DNA片段和核基因ITS片段组合(trn L-trn F+trn M-trn V+trn S-trn G+ITS)的辨别率相同,因此推荐trn S-trn G+ITS作为地黄属植物的标准条形码。此外,利用DNA条形码鉴别物种时,可采用叶绿体DNA片段和核DNA片段组合的方法来提高物种鉴定的成功率。