The Chalcidoidea bush of life: evolutionary history of a massive radiation of minute wasps.

Chalcidoidea are mostly parasitoid wasps that include as many as 500 000 estimated species. Capturing phylogenetic signal from such a massive radiation can be daunting. Chalcidoidea is an excellent example of a hyperdiverse group that has remained recalcitrant to phylogenetic resolution. We combined 1007 exons obtained with Anchored Hybrid Enrichment with 1048 ultra-conserved elements (UCEs) for 433 taxa including all extant families, >95% of all subfamilies, and 356 genera chosen to represent the vast diversity of the superfamily. Going back and forth between the molecular results and our collective knowledge of morphology and biology, we detected bias in the analyses that was driven by the saturation of nucleotide data. Our final results are based on a concatenated analysis of the least saturated exons and UCE datasets (2054 loci, 284 106 sites). Our analyses support an expected sister relationship with Mymarommatoidea. Seven previously recognized families were not monophyletic, so support for a new classification is discussed. Natural history in some cases would appear to be more informative than morphology, as illustrated by the elucidation of a clade of plant gall associates and a clade of taxa with planidial first-instar larvae. The phylogeny suggests a transition from smaller soft-bodied wasps to larger and more heavily sclerotized wasps, with egg parasitism as potentially ancestral for the entire superfamily. Deep divergences in Chalcidoidea coincide with an increase in insect families in the fossil record, and an early shift to phytophagy corresponds with the beginning of the "Angiosperm Terrestrial Revolution". Our dating analyses suggest a middle Jurassic origin of 174 Ma (167.3-180.5 Ma) and a crown age of 162.2 Ma (153.9-169.8 Ma) for Chalcidoidea. During the Cretaceous, Chalcidoidea may have undergone a rapid radiation in southern Gondwana with subsequent dispersals to the Northern Hemisphere. This scenario is discussed with regard to knowledge about the host taxa of chalcid wasps, their fossil record and Earth's palaeogeographic history.

A new genus of Chalcidoidea (Hymenoptera) from Chile with challenging taxonomic position.

An enigmatic chalcid wasp (Hymenoptera: Chalcidoidea) from Chile is described as a new genus and species, Lautaroderus malalcahuello gen. et sp. n. The taxon is classified in the family Eulophidae based on possession of a short, straight calcar and 4-segmented tarsi of female. However, a densely setose basal cell of fore wing, X-shaped pattern of facial grooves, and an incurved anterior margin of the clypeus are similar to representatives of the subfamily Tetracampinae of Tetracampidae. Although L. malalcahuello does not fit readily in the current concept of Eulophidae, it is assigned to this family though not further placed to subfamily or tribe. The proposed classification and possible relationships of the new taxon are discussed.

Afronympha, a new genus of Entedoninae (Hymenoptera: Eulophidae) from the rainforests of Africa.

Afronympha gen. nov. is described for the single known species A. eminpashai sp. nov. collected in the Democratic Republic of the Congo and Uganda. This new genus is similar to some extent to Asecodes, but differs from this and other genera of Entedoninae in having a produced and medially notched anterior margin of the clypeus, and robust mandibles. Similarities and differences between Afronympha and other genera are discussed. The type species, A. eminpashai sp. nov., was collected in degraded mid-altitude rainforest in Kibale National Park, western Uganda and disturbed areas of lowland rainforest in the Ituri region of eastern Democratic Republic of the Congo, suggesting its ecological plasticity under circumstances of habitat transformation.

Review of the species of Pediobius (Hymenoptera, Eulophidae) having extreme dorsal setation and description of a new species from East Africa.

The Afrotropical species of the genus Pediobius (Hymenoptera, Eulophidae) characterized by extreme setation on the dorsal mesosoma are reviewed and grouped into a newly proposed setigerus group. This group is characterized by the broad attachment of the third funicular and first claval segments, so that the funicle is functionally 2-segmented with a 3-segmented clava. Four species are recognized for the group: P. setigerus Kerrich (from Ghana, Ivory Coast, Republic of Guinea, the Democratic Republic of the Congo, Uganda), P. multisetis Bouek (from Ivory Coast), P. sp. indet. aff. multisetis (from Uganda), and P. erinaceus Gumovsky sp. n. (from Uganda). This last species is associated with a leaf miner, and is easily recognizable in having numerous strong bristles on the lateral areas of the mesoscutum, along the notauli, and anterior margin of the mesoscutum, a smooth mesoscutellum with only one pair of thin setae, and the pronotal collar with only 6 thin setae. By contrast, in P. setigerus and P. multisetis the mesoscutellum is sculptured and bears a row of about 20 strong bristles on each side, and the pronotal collar also bears about 20 strong bristles. The differences between P. setigerus and P. multisetis, as well as issues on their biology and type material, are discussed.

Optimized DNA extraction and library preparation for minute arthropods: Application to target enrichment in chalcid wasps used for biocontrol.

Target enrichment is increasingly used for genotyping of plant and animal species or to better understand the evolutionary history of important lineages through the inference of statistically robust phylogenies. Limitations to routine target enrichment are both the complexity of current protocols and low input DNA quantity. Thus, working with tiny organisms such as microarthropods can be challenging. Here, we propose easy to set up optimizations for DNA extraction and library preparation prior to target enrichment. Prepared libraries were used to capture 1,432 ultraconserved elements (UCEs) from microhymenoptera (Chalcidoidea), which are among the tiniest insects on Earth and the most commercialized worldwide for biological control purposes. Results show no correlation between input DNA quantities (1.8-250 ng, 0.4 ng with an extra whole genome amplification step) and the number of sequenced UCEs on an Illumina MiSeq. Phylogenetic inferences highlight the potential of UCEs to solve relationships within the families of chalcid wasps, which has not been achieved so far. The protocol (library preparation + target enrichment) allows processing 96 specimens in five working days, by a single person, without requiring the use of expensive robotic molecular biology platforms, which could help to generalize the use of target enrichment for minute specimens.

New enigmatic species of the genus Pediobius (Hymenoptera, Eulophidae) from Afrotropics, with notes on related genera.

Three new species groups and seven species of the genus Pediobius Walker are described from the Afrotropical realm: the marjoriae group, with P. marjoriae Kerrich (described from Uganda), P. rohombaya Gumovsky sp. n. (from the Central African Republic (CAR), the Democratic Republic of the Congo (DRC), Gabon, Uganda) and P. orungu Gumovsky sp. n. (from Gabon); the afroteres group with P. afroteres Gumovsky sp. n. (from South Africa, Zimbabwe, Malawi, Kenya, Uganda, DRC, Cameroon) and P. kafroteres Gumovsky sp. n. (from Cameroon); the askari group with P. askari Gumovsky sp. n. (from South Africa, Zimbabwe, Kenya, Tanzania, DRC, Ivory Coast) and P. maleficus Gumovsky sp. n. (from Kenya, DRC, CAR, Cameroon, Gabon); and P. nganga Gumovsky sp. n. (from DRC) (not assigned to any group). All these species are characterized by the relatively wide and robust head with elongate and/or narrowed lower face and the antennae attached near or below the lower eye margins. Morphological features of the species, as well as their habitat distribution, are discussed. The comparison of the new species with similar taxa of Entedoninae suggested the following synonymies: Rhynchentedon Girault and Pediobomyia Girault under Pediobius (syn. n.); Bomyiabius frontus Narendran, Pediobomyia budaicus Narendran and Pediobomyia lankicus Narendran under Pediobomyia darwini Girault (syn. n.). The following new combinations are proposed: Pediobius maximus (Girault), P. achterbergi (Gumovsky), P. narendrani (Gumovsky), P. brevicaulis (Hansson), P. canaliculatus (Hansson) and P. darwini (Girault) (comb. n.). Ant parasitoids P. marjoriae and Myrmokata diparoides Boucek are recorded from DRC for the first time.

Review of Afrotropical species of Goetheana Girault (Hymenoptera: Eulophidae), with description of a new species.

Three species of Goetheana Girault (Hymenoptera: Eulophidae, Entedoninae) are recorded from the Afrotropical biogeographic realm: G. shakespearei Girault (cosmopolitan), G. incerta Annecke (Namibia, South Africa, Swaziland, Uganda, Senegal) and G. kobzari Gumovsky sp. n. (South Africa, Uganda, Central African Republic). Goetheana incerta is re-described based on its type series, which is discussed. Males of G. shakespearei and G. incerta are easily separated by antennal scape structure, but females barely differ in morphology. New geographical records and a discussion of morphology of Goetheana are also provided.

A review of genera and described Afrotropical species of Tetracampinae (Hymenoptera: Tetracampidae), with description of a new genus from East Central Africa.

The genera and described Afrotropical species of Tetracampinae Förster (Hymenoptera: Chalcidoidea, Tetracampidae) are reviewed. Despite the vague nature of some generic concepts, eight genera are recognized for the subfamily, i.e. Cassidocida Crawford, Diplesiostigma Girault, Dipriocampe Boucek, Epiclerus Haliday, Foersterella Dalla Torre, Niticampe Boucek, Tetracampe Förster, and the newly described Kilomotoia Gumovsky gen. n. The latter genus is established for Kilomotoia kitoko Gumovsky sp. n., collected from abandoned cultivated areas in the humid forest zone of North East Democratic Republic of Congo (Ituri forest, Ituri Province, environs of Mongbwalu village) and Central Uganda (Mabira forest, environs of Najembe village). Kilomotoia is distinguished from other tetracampine genera by the possession of a flattened dorsum, large abrupt pronotum, relatively long and laterally depressed gaster having gastral tergites II-V with medially emarginate posterior margins, and the hypopygium extending to the apex of gaster as sharply pointed mucro. The four hitherto described Afrotropical tetracampine species are reviewed based on available museum material, with Cassidocida africana Ferrière transferred to Foersterella as F. africana (Ferrière) comb. n., E. agromyzae (Risbec) and E. plectroniae (Risbec) confirmed as species of Epiclerus (as transferred earlier from Foersterella by Boucek 1958 and Boucek & Askew 1968), and Cassidocida orthopterae Risbec transferred to Closterocerus (Eulophidae, Entedoninae) as C. orthopterae (Risbec) comb. n. New diagnostic characters are proposed for the subfamily and some genera, and a comparative character table provided for tetracampine genera.

Discovery of a new species of Caloptilia (Lepidoptera: Gracillariidae) from east and central Africa with its suggested associated host (Gentianales: Rubiaceae) and natural enemies (Hymenoptera: Eulophidae).

A new species of the leaf-mining moth genus Caloptilia (Gracillariidae), C. mwamba sp. nov., suggested to be associated with Cremaspora triflora (Thonn.) K.Schum. (Rubiaceae) is described from east and central Africa. The taxonomic relationships of the new species with its congeners from the Oriental and the Palaearctic regions are discussed. Newly obtained taxonomic and biological data are linked with the DNA barcode workbench in BOLD, providing the molecular, machine-readable identification tag of the new species. New distribution and morphological data for two parasitoid species, Afrotroppopsis risbeci Gumovsky, 2007 and Zaommomentedon newbyi (Kerrich, 1969) (Eulophidae), which were found to be associated with C. mwamba sp. nov., are presented.

Re-description and first host and biology records of Entedon magnificus (Girault & Dodd) (Hymenoptera, Eulophidae), a natural enemy of Gonipterus weevils (Coleoptera, Curculionidae), a pest of Eucalyptus trees.

Entedon magnificus (Girault & Dodd) (Hymenoptera: Eulophidae, Entedoninae) is recorded as a gregarious larval endoparasitoid of Gonipterus weevils (Coleoptera: Curculionidae), significant pests of Eucalyptus trees. Entedon magnificus is re-described and illustrated based on females and males from Australia and Tasmania.

A new species of Trisecodes from the Afrotropics (Hymenoptera, Eulophidae) .

The first Afrotropical species of Trisecodes Delvare & LaSalle, 2000, T. africanum Gumovsky, sp. n., is described from localities in Uganda, Guinea and Cameroon. The new species differs from the type species of the genus, T. agromyzae, which was described from the Neotropical region in Belize, mostly by having longitudinally strigate sculpture on the mesosoma dorsally and by the fore wing being somewhat darkened in its basal three-fifths. A possible Gondwanan origin of the genus is postulated. 

A phylogenetic analysis of the megadiverse Chalcidoidea (Hymenoptera).

Chalcidoidea (Hymenoptera) is extremely diverse with an estimated 500 000 species. We present the first phylogenetic analysis of the superfamily based on both morphological and molecular data. A web-based, systematics workbench mx was used to score 945 character states illustrated by 648 figures for 233 morphological characters for a total of 66 645 observations for 300 taxa. The matrix covers 22 chalcidoid families recognized herein and includes 268 genera within 78 of 83 subfamilies. Morphological data were analysed alone and in combination with molecular data from ribosomal 18S (2105 bp) and 28S D2-D5 expansion regions (1812 bp). Analyses were analysed alone and in combined datasets using implied-weights parsimony and likelihood. Proposed changes in higher classification resulting from the analyses include: (i) recognition of Eriaporidae, revised status; (ii) recognition of Cynipencyrtidae, revised status; (iii) recognition of Azotidae, revised status; (iv) inclusion of Sycophaginae in Agaonidae, revised status; (v) reclassification of Aphelinidae to include Aphelininae, Calesinae, Coccophaginae, Eretmocerinae and Eriaphytinae; (vi) inclusion of Cratominae and Panstenoninae within Pteromalinae (Pteromalidae), new synonymy; (vii) inclusion of Epichrysomallinae in Pteromalidae, revised status. At a higher level, Chalcidoidea was monophyletic, with Mymaridae the sister group of Rotoitidae plus the remaining Chalcidoidea. A eulophid lineage was recovered that included Aphelinidae, Azotidae, Eulophidae, Signiphoridae, Tetracampidae and Trichogrammatidae. Eucharitidae and Perilampidae were monophyletic if Eutrichosomatinae (Pteromalidae) was included, and Eupelmidae was monophyletic if Oodera (Pteromalidae: Cleonyminae) was included. Likelihood recovered a clade of Eupelmidae + (Tanaostigmatidae + (Cynipencyrtus + Encyrtidae). Support for other lineages and their impact on the classification of Chalcidoidea is discussed. Several life-history traits are mapped onto the new phylogeny.

Comparative cytogenetic study on two species of the genus Entedon Dalman, 1820 (Hymenoptera, Eulophidae) using DNA-binding fluorochromes and molecular and immunofluorescent markers.

Karyotypes of Entedon cionobius Thomson, 1878 and Entedon cioni Thomson, 1878 (Hymenoptera: Eulophidae) were studied using DNA-binding ligands with different base specificity (propidium iodide, chromomycin A3, methyl green and DAPI; all these ligands, except for the last one, were used for the first time in parasitic wasps), C-banding, fluorescence in situ hybridization (FISH) with a 45S rDNA probe and 5-methylcytosine immunodetection. Female karyotypes of both species contain five pairs of relatively large metacentric chromosomes and a pair of smaller acrocentric chromosomes (2n = 12). As in many other Hymenoptera, males of both Entedon Dalman, 1820 species have haploid chromosome sets (n = 6). Fluorochrome staining revealed chromosome-specific banding patterns that were similar between the different fluorochromes, except for the CMA3- and PI-positive and DAPI-negative band in the pericentromeric regions of the long arms of both acrocentric chromosomes. The obtained banding patterns were virtually identical in both species and allowed for the identification of each individual chromosome. C-banding revealed a pattern similar to DAPI staining, although centromeric and telomeric regions were stained more intensively using the former technique. FISH detected a single rDNA site in the same position on the acrocentric chromosomes as the bright CMA3-positive band. Immunodetection of 5-methylcytosine that was performed for the first time in the order Hymenoptera revealed 5-methylcytosine-rich sites in the telomeric, centromeric and certain interstitial regions of most of the chromosomes.

A new genus of Entedoninae (Hymenoptera: Eulophidae) from Brazil / Um novo gênero de Entedoninae (Hymenoptera: Eulophidae) no Brasil

Podkova, um novo gênero, descrito do Brasil, para acomodar sua espécie-tipo, P. hirsuta sp. n. O gênero têm uma aparência peculiar, e suas principais características são um corpo pequeno e compacto densamente piloso na porção dorsal da cabeça e mesosoma, incluindo a parte média do propódeo; cabeça com a carena frontal em forma de ferradura, delimitando a cavidade facial larga com as antenas curtas; e a forma da carena occipital estendendo-se posteriormente para baixo, ao longo das margens posteriores dos olhos. Por essas características Podkova assemelha-se ao gênero Alachua Schauff & Boucek, mas diferindo deste por possuir uma forte carena frontal e o prepecto fortemente sobrepsassado pela margemanterior do mesosterno. Possíveis afinidades com outros gêneros de Entedoninae são discutidas.

A new genus, Podkova, is described from Brazil to accommodate its type-species P. hirsuta, sp. n. The genus has a peculiar appearance, the main features of which are the small compact body with coarsely pilose dorsal parts of the head and mesosoma, including median part of propodeum, head with sharp frontal carina of horse-shoe shape, delimiting broad facial cavity with rather short antennae, and a sharp occipital carina stretching posteriorly down along hind margins of eyes. By these features, Podkova resembles the genus Alachua Schauff and Boucek, but differs distinctly by having strong frontal carina on the head and the prepectus hardly overlapped by the anterior margin ofmesosternum. Possible affinities to some other entedonine genera are discussed.