Application of DNA barcoding to the identification of Hymenoptera parasitoids from the soybean aphid (Aphis glycines) in China

Aphis glycines Matsumura is an important pest of soybean in Asia and North America. Hymenoptera parasitoids play a key role in the control of the soybean aphid. The correct identification of parasitoids is a critical step that precedes the assessment of their potential biological control agents. Accurate identification of the majority of the species attacking the soybean aphid often requires elaborate specimen preparation and expert taxonomic knowledge. In this study, we facilitated the identification of soybean aphid parasitoids by applying a DNA barcoding approach following a preliminary morphological identification. We generated DNA sequence data from the mitochondrial COI gene and the D2 region of 28S rDNA to assess the genetic variation within and between parasitoid species emerging from the soybean aphid in China. Fifteen Hymenoptera parasitoid species belonging to 10 genera of five families were identified with little intra‐specific variation (0.09% ± 0.06% for 28S and 0.36% ± 0.18% for COI) and large inter‐specific divergence (30.46% ± 3.42% for 28S and 20.4% ± 1.20% for COI).     

基于28S rDNA D2基因片段与形态特征的矛茧蜂亚科系统发育研究（膜翅目：茧蜂科）

本研究选取矛茧蜂亚科Doryctinae（昆虫纲Insecta：膜翅目Hymenoptera：茧蜂科Braconidae）的6族15属18种做内群,茧蜂科其它7亚科11属11种做外群,首次结合同源核糖体28S rDNA D2基因序列片段和100个形态学和解剖学特征对该亚科进行了系统发育学研究。利用“非圆口类"的小腹茧蜂亚科Microgastrinae为根,以PAUP*4.0和MrBayes 3.0B4软件分别应用最大简约法（MP）和贝叶斯法对矛茧蜂亚科的分子数据和分子数据与非分子数据的结合体进行了运算分析;并以PAUP*4.0对矛茧蜂亚科的28S rDNA D2基因序列片段的碱基组成与碱基替代情况进行了分析。结果表明：矛茧蜂亚科的28S rDNA D2基因序列片段的GC含量在39.33%～48.28%之间变动,而对于碱基替代情况来讲,矛茧蜂亚科各成员间序列变异位点上颠换（transversion）大于转换（transition）。不同的分析算法所产生的系统发育树都表明矛茧蜂亚科是一个界限分明的单系群;在矛茧蜂亚科内,除了吉丁茧蜂族Siragrini为单系群外,其他族（矛茧蜂族Doryctini和方头茧蜂族Hecabolini）都是并系群。对于矛茧蜂亚科内各属之间的相互亲缘关系,不同算法所得的系统发育树的拓扑结构不完全一致,表明矛茧蜂亚科内（属及族）的系统发育关系还有待于进一步研究。     

Phylogeny of the Braconinae (Hymenoptera: Braconidae): A new tribal order!

Generic relationships within the parasitoid wasp subfamily Braconinae are assessed based on a molecular phylogenetic analysis of four gene fragments: mitochondrial cytochrome c oxidase subunit I, 16S rDNA, nuclear 28S D2-D3 rDNA and elongation factor 1-alpha. Our results support the recognition of Aphrastobraconini, Braconini and Coeloidini plus three new tribes: Compsobraconini tribus nov ., Tropobraconini tribus nov . and Virgulibraconini tribus nov . The first of these new tribes is restricted to the New World; the second includes the Old World genera Tropobracon Cameron, Trispinaria Quicke and Grangerbracon Samartsev and Belokobylskij and possibly others, whereas the third comprises the Australian genus Virgulibracon Quicke, plus several other described and undescribed Australian genera. Consistent placement of Amyosoma Viereck with members of the Virgulibraconini tribus nov . is discussed, whereas Amyosoma is left currently unplaced. A preliminary key to tribes is presented, and the characters used to differentiate between Aphrastobraconini and Braconini are revised. Megacoeloides Quicke was never recovered with Coeloides, the type genus of Coeloidini, so it is treated as Braconinae incertae sedis. By combining molecular and morphological traits, nearly all valid genera are assigned to tribes, and the possible relationships of the remainder are discussed. Alienoclypeus Shenefelt, 1978 is synonymised with Atanycolus Förster, 1862 (Atanycolus insolitus (Shenefelt) comb. nov .). Several new genera have been revealed and will be described elsewhere.     

Revision of the orchid bee subgenus Euglossella (Hymenoptera, Apidae), Part I, The decorata species group

Euglossella, one of the most distinctive subgenera of orchid bees of the genus Euglossa, is composed of two characteristic assemblages of species, one of them comprising bees bearing the strongly metallic integument trademark of the genus (viridis species group), and the other consisting of bees with a brown integument shaded with metallic iridescence (decorata species group). Here we provide the first of two parts of a revision of Euglossella, providing diagnostic definitions for the subgenus, the decorata species group, and all the species included therein. Six species are included in the decorata group, one new: Euglossa (Euglossella) aurantia, sp. n.; Euglossa (Euglossella) apiformis Schrottky, resurrected status; Euglossa (Euglossella) decorata Smith, revised status; Euglossa (Euglossella) singularis Mocsáry, revised status; Euglossa (Euglossella) cosmodora Hinojosa-Díaz and Engel; and Euglossa (Euglossella) perpulchra Moure and Schlindwein. Euglossa meliponoides Ducke and Euglossa urarina Hinojosa-Díaz and Engel are newly synonymized under Euglossa decorata, Euglossa decorata ruficauda Cockerell is synonymized under Euglossa singularis, and a neotype is designated for Euglossa apiformis.     

Paurodontella parapitica n. sp. (Nematoda: Hexatylina,Sphaerularioidea) from Kermanshah Province,Western Iran

Paurodontella parapitica n. sp., collected from the rhizosphere of an apple tree in Kermanshah province, western Iran, is described. The new species is characterized by a body length of 505 to 723 µm (females) and 480 to 600 µm (males), lip region continuous by depression; 7 to 8 μm broad, 3 to 4 µm high, stylet length 7 to 9 µm or 1 to 1.3 times the lip region diameter, short postuterine sac of 4 to 6 μm long, lateral fields with five to six incisures; outer incisures crenated and inner incisures weakly crenated, excretory pore situated 90 to 100 µm from anterior end; functional males common in the population, with spicules 24 to 26 μm long. Tail of both sexes similar, almost straight and elongate-conoid. The new species resembles in morphology and morphometrics to four known species of the genus, namely P. apitica, P. minuta, P. myceliophaga, and P. sohailai. The results of phylogenetic analyses based on sequences of D2/D3 expansion region of 28S rRNA gene revealed this genus is polyphyletic in four different clades in Tylenchid.     

One new species and one new record species of Cerapteroceroides Ashmead (Hymenoptera: Encyrtidae) from China

Cerapteroceroides acutiscapus sp. nov. is described as new and Cerapteroceroides manaliensis Hayat and Khan, 2015 is first recorded from China.     

A molecular and morphological reassessment of the phylogeny of the subfamily Ophioninae (Hymenoptera: Ichneumonidae)

The phylogeny of the subfamily Ophioninae (Hymenoptera: Ichneumonidae) is investigated using molecular markers and morphological characters. We analysed the mitochondrial DNA CO1 and the nuclear 28S D2–D3 gene fragments for 74 species of Ophioninae from 25 out of the 32 recognized genera, which collectively represent 98% of described species diversity of the subfamily. Molecular markers were analysed separately and combined, with or without the adjunction of a matrix of 62 morphological characters using Bayesian inference. Our results reveal three distinct lineages, each including one of most speciose genera: Ophion, Enicospilus and Thyreodon. The comparison of the molecular data, and combined molecular plus morphological data led to the definition of the three tribes: Ophionini stat. rev. (Ophion + Alophophion + Rhopalophion + Xylophion + Afrophion); Enicospilini stat. rev. (Enicospilus + Laticoleus + Dicamptus + Hellwigiella); and Thyreodonini tribe nov. (Thyreodon + Dictyonotus + Rhynchophion). The possible association of other genera to one or another of these lineages is discussed. Ophion is a polyphyletic assemblage and requires a further revision to define the delimitation with close genera. The enigmatic Old World genus Skiapus is strongly supported as belonging to the Ophioninae, although its placement within the subfamily is ambiguous as a result of its derived genotype and phenotype. Finally, we propose a biogeographical scenario supported by this phylogeny and based on the limited available fossil data.     

Revision of the Paridris nephtaspecies group (Hymenoptera, Platygastroidea, Platygastridae)

The Paridris nephta group is revised (Hymenoptera: Platygastridae). Fifteen species are described, 14 of which are new: Paridris atroxTalamas, sp. n.(Yunnan Province, China), Paridris bununTalamas, sp. n.(Taiwan), Paridris ferusTalamas, sp. n.(Thailand), Paridris kagemonoTalamas, sp. n.(Japan), Paridris minatorTalamas, sp. n.(Laos, Thailand), Paridris mystaxTalamas, sp. n.(Laos, Thailand), Paridris nephta(Kozlov) (Japan, North Korea, South Korea, Far Eastern Russia), Paridris nilakaTalamas, sp. n.(Thailand), Paridris reptilisTalamas, sp. n.(Taiwan), Paridris rugulosusTalamas, sp. n.(Laos, Vietnam), Paridris solarisTalamas, sp. n.(Laos, Thailand, Vietnam), Paridris teresTalamas, sp. n.(Vietnam), Paridris toketokiTalamas, sp. n.(Taiwan), Paridris verrucosusTalamas, sp. n.(Guangdong Province, China), Paridris yakTalamas, sp. n.(Thailand).     

A taxonomic study of Chinese species of the alberti group of Metaphycus (Hymenoptera,Encyrtidae)

Ten alberti-group species of the genus Metaphycus Mercet from China are reviewed. Six species Metaphycus dorsalis sp. n., Metaphycus chinensis sp. n., Metaphycus wui sp. n., Metaphycus stylatus sp. n., Metaphycus fusiscapus sp. n. and Metaphycus fusiformis sp. n. are described as new to science. Four known species from China are redescribed. A key to the females of the Chinese species is given and photomicrographs are provided to illustrate morphological characters of these species. All specimens unless otherwise specified are deposited in the National Zoological Museum of China Institute of Zoology Chinese Academy of Sciences Beijing.     

A taxonomic study of Chinese species of the insidiosus group of Metaphycus (Hymenoptera,Encyrtidae)

In this paper, twelve insidiosus-group species of the genus Metaphycus Mercet from China are reviewed. Five species, M. corniae sp. n., M. cylindricus sp. n., M. deltoideus sp. n., M. transversus sp. n. and M. yaanensis sp. n., are described as new to science. A key to the females of these species is given to facilitate species recognition. Photomicrographs are provided to illustrate morphological characters of these species. All specimens, unless otherwise specified, are deposited in the National Zoological Museum of China, Institute of Zoology, Chinese Academy of Sciences, Beijing.     

Two new species and one new record species of Rhopus (Hymenoptera: Encyrtidae) from China,with a key to Chinese species

Rhopus antennalis sp. nov. and R. qingdaoensis sp. nov. are described as new to science and R. nigroclavatus (Ashmead, 1902) is first recorded from China. A key to all the known Chinese species of the genus also presented.     

A new species of Ptilomymar (Hymenoptera,Mymaridae) and a key to the described species

Ptilomymar dianensis sp. n. (Hymenoptera, Mymaridae) from southwest China is described and illustrated. A key to the six described species is given. The type specimens are deposited in the insect collections of Northeast Forestry University, China.     

A new species of the Aenictus wroughtonii group (Hymenoptera,Formicidae) from South-East China

A new species of army ant from the Aenictus wroughtonii group is described and illustrated based on the worker caste. Aenictus gutianshanensis Staab, sp. n. is known form a single colony collected in the subtropical mixed evergreen broad-leaved forest of the Gutianshan National Nature Reserve, South-East China. The new species is probably most closely related to A. vieti Jaitrong & Yamane, 2010 known from North Vietnam and Taiwan. It is suggested that the abundant records of A. camposi Wheeler & Chapman, 1925 from East and South-East China should be reevaluated, as they are probably A. gutianshanensis or A. vieti and not A. camposi, which is distributed in Sundaland, the Philippines, and the southernmost part of continental South-East Asia.     

Monograph of the Afrotropical species of Scelio Latreille (Hymenoptera,Platygastridae), egg parasitoids of acridid grasshoppers (Orthoptera,Acrididae)

The genus Scelio is a cosmopolitan and speciose group of solitary parasitoids of the eggs of short-horned grasshoppers (Orthoptera: Acrididae). A number of these hosts are important pests, including plague locusts of the genus Schistocerca. Species of Scelio are recognized as potentially important biological control agents, but this possibility has yet to be fully realized, in part because the species-level taxonomy is still incompletely developed. The species of the pulchripennis group have been recently revised. As a continuation of this effort, here we revise the Afrotropical species of Scelio, excluding the pulchripennis species group. Sixty two (62) species are treated, 48 of which are new. Species are classified into the following species groups: ernstii (12 species, 9 new), howardi (23 species, 19 new), ipomeae (6 species, 5 new), irwini (4 species, 3 new), simoni (3 new species) and walkeri (12 species, 9 new). Keys to species groups and to the species within each group are provided. New species described are: S. albatus Yoder, sp. n., S. aphares Yoder, sp. n., S. apospastos Yoder, sp. n., S. ardelio Yoder, sp. n., S. aurantium Yoder, sp. n., S. balo Valerio & Yoder, sp. n., S. bayanga Yoder, sp. n., S. bubulo Yoder, sp. n., S. cano Yoder, sp. n., S. clypeatus Yoder, sp. n., S. concavus Yoder, sp. n., S. copelandi Yoder, sp. n., S. crepo Yoder, sp. n., S. destico Yoder, sp. n., S. dupondi Yoder, sp. n., S. effervesco Yoder, sp. n., S. erugatus Yoder, sp. n., S. exophthalmus Yoder, sp. n., S. fremo Valerio & Yoder, sp. n., S. gemo Yoder, sp. n., S. grunnio Yoder, sp. n., S. harinhalai Yoder, sp. n., S. igland Yoder, sp. n., S. impostor Yoder, sp. n., S. irwini Yoder, sp. n., S. janseni Yoder, sp. n., S. latro Yoder, sp. n., S. memorabilis Yoder, sp. n., S. modulus Yoder, sp. n., S. mutio Yoder, sp. n., S. ntchisii Yoder, sp. n., S. parkeri Yoder, sp. n., S. phaeoprora Yoder, sp. n., S. pilosilatus Yoder, sp. n., S. pipilo Yoder, sp. n., S. quasiclypeatus Yoder, sp. n., S. retifrons Yoder, sp. n., S. ructo Yoder, sp. n., S. scomma Yoder, sp. n., S. simoni Yoder, sp. n., S. simonolus Yoder, sp. n., S. somaliensis Yoder, sp. n., S. susurro Yoder, sp. n., S. tono Yoder, sp. n., S. transtrum Yoder, sp. n., S. tritus Yoder, sp. n., S. ululo Yoder, sp. n., S. vannoorti Valerio & Yoder, sp. n. The following species are redescribed: S. afer Kieffer, S. chapmani Nixon, S. howardi Crawford, S. ipomeae Risbec, stat. n., S. mauritanicus Risbec, S. philippinensis Ashmead, S. remaudierei Ferrière, S. striatus Priesner,S. taylori Nixon, and S. zolotarevskyi Ferrière. The genus Lepidoscelio Kieffer is treated as a junior synonym of Scelio Latreille, syn. n.; its type species, Lepidoscelio fuscipennis Kieffer, 1905 is transferred to Scelio, renamed Scelio obscuripennis Johnson, nom. n. (preoccupied by Scelio fuscipennis Ashmead, 1887), and redescribed. The following additional species are transferred from Lepidoscelio to Scelio: S. cayennensis (Risbec), comb. n., S. insularis Ashmead, rev. comb., S. luteus (Cameron), comb. n., S. thoracicus Ashmead, rev. comb. Lectotypes are designated for S. africanus Risbec, S. ipomeae Risbec, S. mauritanicus Risbec, S. remaudierei Ferrière, S. sudanensis Ferrière, and S. zolotarevskyi Ferrière. Scelio gaudens Nixon is a junior synonym of Scelio striatus Priesner, syn. n.; Scelio africanus Risbec and Scelio clarus Fouts are both junior synonyms of Scelio afer Kieffer, syn. n.; Scelio sudanensis Ferrière and Scelio cheops Nixon are both junior synonyms of Scelio zolotarevskyi Ferrière, syn. n.; Scelio cahirensis Priesner is a junior synonym of Scelio mauritanicus Risbec, syn. n. The name Scelio chapmanni Nixon is an incorrect original spelling, requiring an emendation to S. chapmani. Digital versions of the identification keys are available at http://www.waspweb.org/Platygastroidea/Keys/index.htm     

An integrative approach to characterize cryptic species in the Thoracostoma trachygaster Hope, 1967 complex (Nematoda: Leptosomatidae)

Nematode diversity may seriously be underestimated when taking into account cryptic speciation. Thoracostoma trachygaster is commonly found in kelp holdfasts along the California coastline and was recently shown to consist of at least two distinct molecular clades (I and II). Here, we provide detailed morphological analysis of both clades, based on measurements taken from video vouchers of respectively eight and 16 individuals from the previous study, as well as 80 newly collected specimens from four Californian beaches. The latter were vouchered, measured, and then subjected to molecular analyses of the mitochondrial cytochrome oxidase c subunit I (COI) gene, and the ribosomal D2D3 and internal transcribed spacer (ITS) regions. This integrative approach shows that the three molecular clades are phylogenetically and morphologically distinct species, but a combination of morphological characters is needed to distinguish them. Two new species, Thoracostoma fatimae sp. nov. and Thoracostoma igniferum sp. nov. , are identified and described. The spicule length of T. fatimae sp. nov. is significantly shorter than that of T. trachygaster. Thoracostoma igniferum sp. nov. can be distinguished by the irregular posterior edge of the cephalic capsule and the two internal subdorsal tropis‐like projections in the wall of the cephalic capsule, which are lacking in T. fatimae sp. nov. and T. trachygaster. © 2012 The Linnean Society of London, Zoological Journal of the Linnean Society, 2012, 164 , 18–35.     

Marphysa (Eunicidae,polychaete, Annelida) species of the Sanguinea group from Australia,with comments on pseudo‐cryptic species

The genus Marphysa is widely collected for bait by recreational fishermen in Australia, and yet the number of species remains unresolved. A new species is described together with additional information on a previously described species. This study reveals that while species of Marphysa superficially resemble one another, detailed studies on the distribution of different types of chaetae along the body, together with molecular studies, uncover species which may co‐occur. Correct identification is critical for management of these largely intertidal species. This study also highlights the increasing awareness of the presence of pseudo‐cryptic species in polychaetes.     

Molecular phylogeny of the nematode genus Longidorus (Nematoda: Longidoridae) with description of three new species

The phylum Nematoda includes the genus Longidorus, a remarkable group of invertebrates that are polyphagous root‐ectoparasites of many plants including various agricultural crops and trees. Damage is caused by direct feeding on root cells as well as by transmitting nepoviruses. Species discrimination in Longidorus is complicated by phenotypic plasticity (intraspecific variability and minor interspecific differences) leading to potential misidentification. We conducted nematode surveys in cultivated and natural environments in southern Spain that detected 11 species of Longidorus. We developed a comparative study amongst these related species by considering morphological and morphometric features together with molecular data from nuclear ribosomal RNA genes [D2‐D3 expansion segments of large ribosomal subunit (28S), internal transcribed spacer 1 (ITS1), and partial small ribosomal subunit (18S)]. The results of our molecular and phylogenetic analyses confirmed the morphological hypotheses and allowed the delimitation and discrimination of three new species of the genus, described herein as Longidorus baeticus sp. nov. , Longidorus oleae sp. nov. , and Longidorus andalusicus sp. nov. , and eight known species (Longidorus alvegus, Longidorus crataegi, Longidorus fasciatus, Longidorus intermedius, Longidorus iuglandis, Longidorus magnus, Longidorus rubi, and Longidorus vineacola). Phylogenetic analyses of Longidorus spp. based on the three molecular markers resulted in a general consensus of these species grouping, as lineages were maintained for the majority of species (i.e. species with a conoid‐rounded lip region, amphidial fovea asymmetrically bilobed, female tail bluntly rounded), but not in some others (i.e. positions of L. crataegi, L. intermedius, and L. rubi were quite variable). To date, this is the most complete phylogenetic analysis for Longidorus and Paralongidorus species, with the highest number of species included. No correspondence between phylogenetic trees and morphological characters was found for ribosomal markers, with the exception of amphidial shape. Thus, polyphasic identification, based on integration of molecular analysis with morphology, is a tool beyond doubt in Longidorus identification. © 2013 The Linnean Society of London