Mitogenome architecture supports the non-monophyly of the cosmopolitan parasitoid wasp subfamily Doryctinae (Hymenoptera: Braconidae) recovered by nuclear and mitochondrial phylogenomics.

Mitochondrial DNA gene organisation is an important source of phylogenetic information for various metazoan taxa at different evolutionary timescales, though this has not been broadly tested for all insect groups nor within a phylogenetic context. The cosmopolitan subfamily Doryctinae is a highly diverse group of braconid wasps mainly represented by ectoparasitoids of xylophagous beetle larvae. Previous molecular studies based on Sanger and genome-wide (ultraconserved elements, UCE; and mitochondrial genomes) sequence data have recovered a non-monophyletic Doryctinae, though the relationships involved have always been weakly supported. We characterised doryctine mitogenomes and conducted separate phylogenetic analyses based on mitogenome and UCE sequence data of ~100 representative doryctine genera to assess the monophyly and higher-level classification of the subfamily. We identified rearrangements of mitochondrial transfer RNAs (tRNAs) that support a non-monophyletic Doryctinae consisting of two separate non-related clades with strong geographic structure ('New World' and 'Old World' clades). This geographic structure was also consistently supported by the phylogenetic analyses preformed with mitogenome and UCE sequence data. These results highlight the utility of the mitogenome gene rearrangements as a potential source of phylogenetic information at different evolutionary timescales.

Evaluating the use of common grasses by the wheat stem sawfly (Hymenoptera: Cephidae) and its native parasitoids in rangeland and Conservation Reserve Program grasslands.

The wheat stem sawfly, Cephus cinctus Norton (Hymenoptera: Cephidae), is a major pest of wheat (Triticum aestivum L., Poales: Poaceae) across the northern Great Plains of North America. Cephus cinctus has a wide host range, attacking numerous wild grasses and cultivated cereals in crop and grassland habitats, where it is, in turn, attacked by 2 native braconid parasitoids. Quantitative assessments of C. cinctus infestation and parasitism levels in different grass species across the full spectrum of available hosts are important in assessing the extent to which grasslands, or specific constituent grass species, may be reservoirs of pests or parasitoids moving into wheat. We quantified infestation and parasitism levels in over 25,000 stems collected from 17 grass species and wheat spanning 35 sites in central Montana, United States, over 2 yr. Infestation levels in 5 grass species, primarily wheatgrasses, were high (38%-65%) and similar to the levels observed in wheat (55%). In contrast, the majority of grass species (12 of 17) had significantly lower levels of infestation (<10%), suggesting that most grasses are not important reservoirs of C. cinctus. Parasitism levels in highly infested wheatgrasses were, on average, 3 times higher than those in cultivated wheat, suggesting that these grasses could provide important conservation habitat for parasitoids. Future work examining the relative performance of pests and parasitoids in these grasses will be important in gauging their relative value as plant materials to bolster parasitoid conservation in reseeded grassland habitats.

Key innovations and the diversification of Hymenoptera.

The order Hymenoptera (wasps, ants, sawflies, and bees) represents one of the most diverse animal lineages, but whether specific key innovations have contributed to its diversification is still unknown. We assembled the largest time-calibrated phylogeny of Hymenoptera to date and investigated the origin and possible correlation of particular morphological and behavioral innovations with diversification in the order: the wasp waist of Apocrita; the stinger of Aculeata; parasitoidism, a specialized form of carnivory; and secondary phytophagy, a reversal to plant-feeding. Here, we show that parasitoidism has been the dominant strategy since the Late Triassic in Hymenoptera, but was not an immediate driver of diversification. Instead, transitions to secondary phytophagy (from parasitoidism) had a major influence on diversification rate in Hymenoptera. Support for the stinger and the wasp waist as key innovations remains equivocal, but these traits may have laid the anatomical and behavioral foundations for adaptations more directly associated with diversification.

Mitochondrial phylogenomics and mitogenome organization in the parasitoid wasp family Braconidae (Hymenoptera: Ichneumonoidea).

BACKGROUND: Mitochondrial (mt) nucleotide sequence data has been by far the most common tool employed to investigate evolutionary relationships. While often considered to be more useful for shallow evolutionary scales, mt genomes have been increasingly shown also to contain valuable phylogenetic information about deep relationships. Further, mt genome organization provides another important source of phylogenetic information and gene reorganizations which are known to be relatively frequent within the insect order Hymenoptera. Here we used a dense taxon sampling comprising 148 mt genomes (132 newly generated) collectively representing members of most of the currently recognised subfamilies of the parasitoid wasp family Braconidae, which is one of the largest radiations of hymenopterans. We employed this data to investigate the evolutionary relationships within the family and to assess the phylogenetic informativeness of previously known and newly discovered mt gene rearrangements. RESULTS: Most subfamilial relationships and their composition obtained were similar to those recovered in a previous phylogenomic study, such as the restoration of Trachypetinae and the recognition of Apozyginae and Proteropinae as valid braconid subfamilies. We confirmed and detected phylogenetic signal in previously known as well as novel mt gene rearrangements, including mt rearrangements within the cyclostome subfamilies Doryctinae and Rogadinae. CONCLUSIONS: Our results showed that both the mt genome DNA sequence data and gene organization contain valuable phylogenetic signal to elucidate the evolution within Braconidae at different taxonomic levels. This study serves as a basis for further investigation of mt gene rearrangements at different taxonomic scales within the family.

Phylogenomics of braconid wasps (Hymenoptera, Braconidae) sheds light on classification and the evolution of parasitoid life history traits.

The parasitoid lifestyle is largely regarded as a key innovation that contributed to the evolutionary success and extreme species richness of the order Hymenoptera. Understanding the phylogenetic history of hyperdiverse parasitoid groups is a fundamental step in elucidating the evolution of biological traits linked to parasitoidism. We used a genomic-scale dataset based on ultra-conserved elements and the most comprehensive taxon sampling to date to estimate the evolutionary relationships of Braconidae, the second largest family of Hymenoptera. Based on our results, we propose Braconidae to comprise 41 extant subfamilies, confirmed a number of subfamilial placements and proposed subfamily-level taxonomic changes, notably the restoration of Trachypetinae stat. rev. and Masoninae stat. rev. as subfamilies of Braconidae, confirmation that Apozyx penyai Mason belongs in Braconidae placed in the subfamily Apozyginae and the recognition of Ichneutinae sensu stricto and Proteropinae as non-cyclostome subfamilies robustly supported in a phylogenetic context. The correlation between koinobiosis with endoparasitoidism and idiobiosis with ectoparasitoidism, long thought to be an important aspect in parasitoid life history, was formally tested and confirmed in a phylogenetic framework. Using ancestral reconstruction methods based on both parsimony and maximum likelihood, we suggest that the ancestor of the braconoid complex was a koinobiont endoparasitoid, as was that of the cyclostome sensu lato clade. Our results also provide strong evidence for one transition from endo- to ectoparasitoidism and three reversals back to endoparasitoidism within the cyclostome sensu stricto lineage. Transitions of koino- and idiobiosis were identical to those inferred for endo- versus ectoparasitoidism, except with one additional reversal back to koinobiosis in the small subfamily Rhysipolinae.

A Coordinated Sampling and Identification Methodology for Larval Parasitoids of Spotted-Wing Drosophila.

We provide recommendations for sampling and identification of introduced larval parasitoids of spotted-wing drosophila, Drosophila suzukii (Matsumura) (Diptera: Drosophilidae). These parasitoids are either under consideration for importation (aka classical) biological control introductions, or their adventive (presumed to have been accidentally introduced) populations have recently been discovered in North America and Europe. Within the context of the ecology of D. suzukii and its parasitoids, we discuss advantages and disadvantages of estimating larval parasitism levels using different methods, including naturally collected fruit samples and sentinel baits. For most situations, we recommend repeated sampling of naturally occurring fruit rather than using sentinel baits to monitor seasonal dynamics of host plant-Drosophila-parasitoid associations. We describe how to separate Drosophilidae puparia from host fruit material in order to accurately estimate parasitism levels and establish host-parasitoid associations. We provide instructions for identification of emerging parasitoids and include a key to the common families of parasitoids of D. suzukii. We anticipate that the guidelines for methodology and interpretation of results that we provide here will form the basis for a large, multi-research team sampling effort in the coming years to characterize the biological control and nontarget impacts of accidentally and intentionally introduced larval parasitoids of D. suzukii in several regions of the world.

DROP: Molecular voucher database for identification of Drosophila parasitoids.

Molecular identification is increasingly used to speed up biodiversity surveys and laboratory experiments. However, many groups of organisms cannot be reliably identified using standard databases such as GenBank or BOLD due to lack of sequenced voucher specimens identified by experts. Sometimes a large number of sequences are available, but with too many errors to allow identification. Here, we address this problem for parasitoids of Drosophila by introducing a curated open-access molecular reference database, DROP (Drosophila parasitoids). Identifying Drosophila parasitoids is challenging and poses a major impediment to realize the full potential of this model system in studies ranging from molecular mechanisms to food webs, and in biological control of Drosophila suzukii. In DROP, genetic data are linked to voucher specimens and, where possible, the voucher specimens are identified by taxonomists and vetted through direct comparison with primary type material. To initiate DROP, we curated 154 laboratory strains, 856 vouchers, 554 DNA sequences, 16 genomes, 14 transcriptomes, and six proteomes drawn from a total of 183 operational taxonomic units (OTUs): 114 described Drosophila parasitoid species and 69 provisional species. We found species richness of Drosophila parasitoids to be heavily underestimated and provide an updated taxonomic catalogue for the community. DROP offers accurate molecular identification and improves cross-referencing between individual studies that we hope will catalyse research on this diverse and fascinating model system. Our effort should also serve as an example for researchers facing similar molecular identification problems in other groups of organisms.

Deployment of Aggregation-Sex Pheromones of Longhorned Beetles (Coleoptera: Cerambycidae) Facilitates the Discovery and Identification of their Parasitoids.

Longhorned beetles (Coleoptera: Cerambycidae) include many species that are among the most damaging pests of managed and natural forest ecosystems worldwide. Many species of cerambycids use volatile chemical signals (i.e., pheromones) to locate mates. Pheromones are often used by natural enemies, including parasitoids, to locate hosts and therefore can be useful tools for identifying host-parasitoid relationships. In two field experiments, we baited linear transects of sticky traps with pheromones of cerambycid beetles in the subfamily Cerambycinae. Enantiomeric mixtures of four linear alkanes or four linear alkanes and a ketol were tested separately to evaluate their attractiveness to hymenopteran parasitoids. We hypothesized that parasitoids would be attracted to these pheromones. Significant treatment effects were found for 10 species of parasitoids. Notably, Wroughtonia ligator (Say) (Hymenoptera: Braconidae) was attracted to syn-hexanediols, the pheromone constituents of its host, Neoclytus acuminatus acuminatus (F.) (Coleoptera: Cerambycidae). Location and time of sampling also significantly affected responses for multiple species of parasitoids. These findings contribute to the basic understanding of cues that parasitoids use to locate hosts and suggest that pheromones can be used to hypothesize host relationships between some species of cerambycids and their parasitoids. Future work should evaluate response by known species of parasitoids to the complete blends of pheromones used by the cerambycids they attack, as well as other odors that are associated with host trees of cerambycids.

Cerambycid Communities and Their Associated Hymenopteran Parasitoids From Major Hardwood Trees in Delaware: Implications for Biocontrol of Invasive Longhorned Beetles.

Cerambycidae provide important ecological services in forests yet cause economic damage when they infest living trees. Parasitoids can regulate woodborer populations, providing considerable control of pest cerambycids. Identifying parasitoids of native cerambycids may be useful in managing cerambycid outbreaks and aid in new-association biocontrol of exotic invasive cerambycids. We investigated Cerambycidae and associated hymenopteran parasitoid communities infesting Acer rubrum, Pinus virginiana, and Carya tomentosa from a forest in Delaware from 2005 to 2012. Cerambycid abundance, diversity, and richness, as well as parasitoid abundance, were measured by collecting trees in different conditions: felled, girdled, and naturally infested. Effect of edge or interior red maple on cerambycid abundance, diversity, and richness was examined. Over 14,500 cerambycids of 56 species and 38 genera were collected during the 7-yr period. Eleven species represented 95% of all cerambycids collected. Treatment only affected red maple, showing increased cerambycid richness and diversity from naturally infested trees. Cerambycid richness and diversity were two times greater on hickory than other species when combining girdled and felled treatments. Over 19,000 parasitic Hymenoptera of 12 families emerged from woodborer-infested wood with >70% of individuals belonging to Braconidae. Thirteen known species, and two unknown species, of Braconidae were identified from a subsample of 495 specimens; Ontsira mellipes (Ashmead) (Hymenoptera: Braconidae) and Rhoptrocentrus piceus Marshall (Hymenoptera: Braconidae) were the most abundant. This study provides fundamental information on native parasitoids associated with Cerambycidae, including cerambycid larval host associations. Parasitoids identified herein should be investigated for potential adaptation to invasive Cerambycidae to benefit invasive woodborer management.

Two new species of Coelinius Nees (Hymenoptera, Braconidae, Alysiinae) from Mexico.

Two new species, Coelinius carmenae Figueroa, Sánchez, and Kula, n. sp. and Coelinius danielae Figueroa, Sánchez, and Kula, n. sp., from Mexico are described and illustrated. These are the first records of Coelinius sensu Wharton (1994) reported from Mexico. Four Nearctic species, Coelinius mahackemoi (Viereck), stat. rev., Coelinius meromyzae Forbes, stat. rev., Coelinius nigripes Ashmead, stat. rev., and Coelinius occom (Viereck), stat. rev. are returned to Coelinius Nees from Coelinidea Viereck. Coelinius longulus Ashmead, stat. rev. is returned from Lepton Zetterstedt, 1838 nec Turton, 1822 to Coelinius. Coelinius linearis (Provancher), n. comb. is transferred from Coelinidea to Coelinius.

Two new species of Anisocyrta Foerster (Hymenoptera: Braconidae: Alysiinae) from China.

Two new species of the genus Anisocyrta Foerster, 1863, from China are described and illustrated: Anisocyrta xiaoliyaoae Yao, sp. n. and Anisocyrta cvanachterbergi Yao, sp. n. Anisocyrta is reported from China for the first time. A key to the Palearctic species of the genus Anisocyrta is provided based on the keys of van Achterberg (1986) and Belokobylskij (1998). Type specimens of the two new species are deposited in the Beneficial Insects Institute Collection, Fujian Agriculture and Forestry University, Fuzhou, China (BIIC).

Phylogenomic Insights into the Evolution of Stinging Wasps and the Origins of Ants and Bees.

The stinging wasps (Hymenoptera: Aculeata) are an extremely diverse lineage of hymenopteran insects, encompassing over 70,000 described species and a diversity of life history traits, including ectoparasitism, cleptoparasitism, predation, pollen feeding (bees [Anthophila] and Masarinae), and eusociality (social vespid wasps, ants, and some bees) [1]. The most well-studied lineages of Aculeata are the ants, which are ecologically dominant in most terrestrial ecosystems [2], and the bees, the most important lineage of angiosperm-pollinating insects [3]. Establishing the phylogenetic affinities of ants and bees helps us understand and reconstruct patterns of social evolution as well as fully appreciate the biological implications of the switch from carnivory to pollen feeding (pollenivory). Despite recent advancements in aculeate phylogeny [4-11], considerable uncertainty remains regarding higher-level relationships within Aculeata, including the phylogenetic affinities of ants and bees [5-7]. We used ultraconserved element (UCE) phylogenomics [7, 12] to resolve relationships among stinging-wasp families, gathering sequence data from >800 UCE loci and 187 samples, including 30 out of 31 aculeate families. We analyzed the 187-taxon dataset using multiple analytical approaches, and we evaluated several alternative taxon sets. We also tested alternative hypotheses for the phylogenetic positions of ants and bees. Our results present a highly supported phylogeny of the stinging wasps. Most importantly, we find unequivocal evidence that ants are the sister group to bees+apoid wasps (Apoidea) and that bees are nested within a paraphyletic Crabronidae. We also demonstrate that taxon choice can fundamentally impact tree topology and clade support in phylogenomic inference.

A new species of Acanthocaudus Smith (Braconidae: Aphidiinae), with a key to species and new host and distribution records for aphidiines associated with Silphium perfoliatum L. (Asterales: Asteraceae).

A new species, Acanthocaudus bicolor Kula (Braconidae: Aphidiinae), from the Nearctic Region is described and differentiated from all other species of Acanthocaudus Smith. Acanthocaudus schlingeri Muesebeck, 1958 is synonymized with Acanthocaudus tissoti (Smith, 1944). A key to the species of Acanthocaudus is provided. The following aphidiines are reported as parasitoids of aphids on Silphium perfoliatum L. (cup plant) for the first time: Aphidius (Aphidius) ohioensis ex Uroleucon (Uroleucon) cf. rudbeckiae (Fitch), Aphidius (Aphidius) polygonaphis (Fitch) ex Uroleucon sp., Praon pequodorum Viereck ex undetermined aphids, and Praon simulans (Provancher) ex undetermined aphids. Aphidius (A.) ohioensis and P. simulans are reported from Indiana and South Dakota for the first time. Acanthocaudus tissoti and P. pequodorum are reported from Indiana for the first time.

Four new species of Tanycarpa (Hymenoptera, Braconidae, Alysiinae) from the Palaearctic Region and new records of species from China.

Four new species of Tanycarpa (Hymenoptera, Braconidae, Alysiinae), T. gymnonotum Yao sp. n., T. similis Yao sp. n., T. areolata Yao sp. n., and T. lineata Yao sp. n., are described from the Palaearctic Region of China, and T. chors Belokobylskij is newly recorded from China. Significant range extensions are given for T. bicolor (Nees von Esenbeck), T. gracilicornis (Nees von Esenbeck), and T. mitis Stelfox. A key to the Palaearctic species of Tanycarpa is provided.

A new species in the newly recorded genus Trachyusa (Hymenoptera: Braconidae: Alysiinae) from China.

Trachyusa whartoni Yao sp. nov. from Dalian, Liaoning, China is described and illustrated. Trachyusa is a newly recorded genus in China. A key to the species of Trachyusa is provided based on the keys of Belokobylskij (1998), Papp (1967), and van Achterberg and O'Connor (1990). Type specimens of the new species are deposited in the Beneficial Insects Institute, Fujian Agriculture and Forestry University, Fuzhou, China.

A new species and new distribution records for Braconidae from Mountain Lake Biological Station in southwestern Virginia and a redescription of Pentapleura foveolata Viereck.

One new species of Alysiinae, Coelinius wrayi Kula, is described. Pentapleurafoveolata Viereck, also in Alysiinae and previously known only from a male collected at the type locality in Connecticut, is redescribed based on six females and two males collected at Mountain Lake Biological Station (MLBS) in Virginia. Morphological variation for Alysia (Alysia) salebrosa Wharton is discussed given variation observed in specimens from MLBS. Sixty-two species of Braconidae collected at MLBS in August of 2009 are listed along with 12 species reported previously from MLBS. Host use for the 62 species is discussed; 31 are in Alysiinae or Opiinae, subfamilies that exclusively contain parasitoids of cyclorrhaphous flies. Fifteen species are reported from Virginia for the first time. Problematic couplets in a key to species of Spathius Nees for North America (Marsh & Strazanac 2009) are discussed using specimens from MLBS.