Mind the Outgroup and Bare Branches in Total-Evidence Dating: a Case Study of Pimpliform Darwin Wasps (Hymenoptera, Ichneumonidae).

Taxon sampling is a central aspect of phylogenetic study design, but it has received limited attention in the context of total-evidence dating, a widely used dating approach that directly integrates molecular and morphological information from extant and fossil taxa. We here assess the impact of commonly employed outgroup sampling schemes and missing morphological data in extant taxa on age estimates in a total-evidence dating analysis under the uniform tree prior. Our study group is Pimpliformes, a highly diverse, rapidly radiating group of parasitoid wasps of the family Ichneumonidae. We analyze a data set comprising 201 extant and 79 fossil taxa, including the oldest fossils of the family from the Early Cretaceous and the first unequivocal representatives of extant subfamilies from the mid-Paleogene. Based on newly compiled molecular data from ten nuclear genes and a morphological matrix that includes 222 characters, we show that age estimates become both older and less precise with the inclusion of more distant and more poorly sampled outgroups. These outgroups not only lack morphological and temporal information but also sit on long terminal branches and considerably increase the evolutionary rate heterogeneity. In addition, we discover an artifact that might be detrimental for total-evidence dating: "bare-branch attraction," namely high attachment probabilities of certain fossils to terminal branches for which morphological data are missing. Using computer simulations, we confirm the generality of this phenomenon and show that a large phylogenetic distance to any of the extant taxa, rather than just older age, increases the risk of a fossil being misplaced due to bare-branch attraction. After restricting outgroup sampling and adding morphological data for the previously attracting, bare branches, we recover a Jurassic origin for Pimpliformes and Ichneumonidae. This first age estimate for the group not only suggests an older origin than previously thought but also that diversification of the crown group happened well before the Cretaceous-Paleogene boundary. Our case study demonstrates that in order to obtain robust age estimates, total-evidence dating studies need to be based on a thorough and balanced sampling of both extant and fossil taxa, with the aim of minimizing evolutionary rate heterogeneity and missing morphological information. [Bare-branch attraction; ichneumonids; fossils; morphological matrix; phylogeny; RoguePlots.].

Phylogenomics of Ichneumonoidea (Hymenoptera) and implications for evolution of mode of parasitism and viral endogenization.

Ichneumonoidea is one of the most diverse lineages of animals on the planet with >48,000 described species and many more undescribed. Parasitoid wasps of this superfamily are mostly beneficial insects that attack and kill other arthropods and are important for understanding diversification and the evolution of life history strategies related to parasitoidism. Further, some lineages of parasitoids within Ichneumonoidea have acquired endogenous virus elements (EVEs) that are permanently a part of the wasp's genome and benefit the wasp through host immune disruption and behavioral control. Unfortunately, understanding the evolution of viral acquisition, parasitism strategies, diversification, and host immune disruption mechanisms, is deeply limited by the lack of a robust phylogenetic framework for Ichneumonoidea. Here we design probes targeting 541 genes across 91 taxa to test phylogenetic relationships, the evolution of parasitoid strategies, and the utility of probes to capture polydnavirus genes across a diverse array of taxa. Phylogenetic relationships among Ichneumonoidea were largely well resolved with most higher-level relationships maximally supported. We noted codon use biases between the outgroups, Braconidae, and Ichneumonidae and within Pimplinae, which were largely solved through analyses of amino acids rather than nucleotide data. These biases may impact phylogenetic reconstruction and caution for outgroup selection is recommended. Ancestral state reconstructions were variable for Braconidae across analyses, but consistent for reconstruction of idiobiosis/koinobiosis in Ichneumonidae. The data suggest many transitions between parasitoid life history traits across the whole superfamily. The two subfamilies within Ichneumonidae that have polydnaviruses are supported as distantly related, providing strong evidence for two independent acquisitions of ichnoviruses. Polydnavirus capture using our designed probes was only partially successful and suggests that more targeted approaches would be needed for this strategy to be effective for surveying taxa for these viral genes. In total, these data provide a robust framework for the evolution of Ichneumonoidea.

The potential of the solitary parasitoid Microctonus brassicae for the biological control of the adult cabbage stem flea beetle, Psylliodes chrysocephala.

The cabbage stem flea beetle (CSFB), Psylliodes chrysocephala L. (Coleoptera: Chrysomelidae), is a major pest of oilseed rape, Brassica napus L. (Brassicaceae), within the UK and continental Europe. Following the withdrawal of many broad-spectrum pesticides, most importantly neonicotinoids, and with increased incidence of pyrethroid resistance, few chemical control options remain, resulting in the need for alternative pest management strategies. We identified the parasitoid wasp Microctonus brassicae (Haeselbarth) (Hymenoptera: Braconidae) within CSFB collected from three independent sites in Norfolk, UK. Parasitism of adult CSFB was confirmed, and wasp oviposition behaviour was described. Moreover, we show that within captive colonies parasitism rates are sufficient to generate significant biological control of CSFB populations. A sequence of the M. brassicae mitochondrial cytochrome oxidase 1 (MT-CO1) gene was generated for rapid future identification. Moroccan specimens of Microctonus aethiopoides (Loan), possessing 90% sequence similarity, were the closest identified sequenced species. This study represents the first description published in English of this parasitoid of the adult cabbage stem flea beetle.

Twenty seven new species of Orthocentrus (Hymenoptera: Ichneumonidae; Orthocentrinae) with a key to the Neotropical species of the genus.

We describe 27 new species of parasitoid wasps of the genus Orthocentrus (Ichneumonidae: Orthocentrinae) from the Neotropical region, where previously only one species of the genus (O. insularis Ashmead) was known, and provide a key to all described Orthocentrus species of the region. Based on previous studies and additional material that we have seen from the region, describing these species is only an initial attempt to document the Neotropical orthocentrine fauna.

First record of the genus Aplomerus Provancher, 1886 (Hymenoptera: Ichneumonidae: Xoridinae) from the Oriental region, with descriptions of two new species.

The genus Aplomerus Provancher, 1886 (Hymenoptera, Ichneumonidae) is a small genus of the subfamily Xoridinae. Two new species from Asia, A. orientalis Varga & Reshchikov sp. n. from Thailand and A. phamae Broad sp. n. from Vietnam, are described and illustrated. An identification key for Asian species is provided. These are the first records of the genus from the Oriental region and increases the known number of Aplomerus species to eight, with a disjunct distribution of North America, Japan and south-east Asia.

The neotropical species of Xanthopimpla Saussure (Hymenoptera: Ichneumonidae: Pimplinae).

Xanthopimpla Saussure, 1892 is one of the largest and best studied genera of the family Ichneumonidae. It is most species rich in the Oriental and Afrotropical regions with only a few species occurring in Central and South America. The present study reviews the Neotropical species of the genus including descriptions of four new species from Amazonia and Northeast South America. We define a new species group: the amazonica species-group, to accommodate the following five species: X. amazonica Gómez, Sääksjärvi & Veijalainen, X. guianensis Gómez & Sääksjärvi sp. n., X. jussilai Veijalainen, Sääksjärvi & Broad, X. pucallpensis Gómez & Sääksjärvi sp. n. and X. vidali Gómez sp. n. The aurita species-group, which had hitherto been regarded as the only species-group in the Neotropical region, is currently represented by five species: X. allpahuaya Gómez & Sääksjärvi sp. n., X. aurita Krieger, X. craspedoptera Krieger, X. rhabdomera Townes and X. spiloptera Krieger. The Andean species X. peruana Krieger is established as an unplaced species outside of the amazonica and aurita species-groups. A key to Neotropical species-groups and species of Xanthopimpla is provided. Xanthopimpla aurita is recorded for the first time from Ecuador and Colombia and its extensive distribution is discussed. Xanthopimpla amazonica, X. craspedoptera and X. jussilai are recorded for the first time from Brazil; X. amazonica is recorded for the first time from French Guiana; X. spiloptera is recorded for the first time from French Guiana and Peru, and X. rhabdomera is recorded for the first time from Peru. 

The genome sequence of a braconid wasp, Aleiodes leptofemur van Achterberg & Shaw, 2016.

We present a genome assembly from an individual male Aleiodes leptofemur (braconid wasp; Arthropoda; Insecta; Hymenoptera; Braconidae). The genome sequence spans 271.20 megabases. Most of the assembly is scaffolded into 15 chromosomal pseudomolecules. The mitochondrial genome has also been assembled and is 32.28 kilobases in length.

The genome sequence of the Dark Crimson Underwing moth, Catocala sponsa Linnaeus, 1767.

We present a genome assembly from an individual female Catocala sponsa (the Dark Crimson Underwing; Arthropoda; Insecta; Lepidoptera; Erebidae). The genome sequence spans 803.70 megabases. Most of the assembly is scaffolded into 32 chromosomal pseudomolecules, including the Z and W sex chromosomes. The mitochondrial genome has also been assembled and is 15.57 kilobases in length. Gene annotation of this assembly on Ensembl identified 13,493 protein-coding genes.

The genome sequence of an ichneumonid wasp, Exephanes ischioxanthus (Gravenhorst, 1829).

We present a genome assembly from an individual male Exephanes ischioxanthus (an ichneumonid wasp; Arthropoda; Insecta; Hymenoptera; Ichneumonidae). The genome sequence is 284.0 megabases in span. Most of the assembly is scaffolded into 12 chromosomal pseudomolecules. The mitochondrial genome has also been assembled and is 19.43 kilobases in length.

The genome sequence of the Marbled Minor moth, Oligia strigilis (Linnaeus, 1758).

We present a genome assembly from an individual male Oligia strigilis (Marbled Minor; Arthropoda; Insecta; Lepidoptera; Noctuidae). The genome sequence is 626.1 megabases in span. Most of the assembly is scaffolded into 31 chromosomal pseudomolecules, including the Z sex chromosome. The mitochondrial genome has also been assembled and is 15.35 kilobases in length.

The genome sequence of an ichneumonid wasp, Hyposoter dolosus (Gravenhorst, 1829).

We present a genome assembly from an individual male Hyposoter dolosus (ichneumonid wasp; Arthropoda; Insecta; Hymenoptera; Ichneumonidae). The genome sequence spans 222.70 megabases. Most of the assembly is scaffolded into 12 chromosomal pseudomolecules. The mitochondrial genome has also been assembled and is 27.89 kilobases in length.

The genome sequence of the moss carder bee, Bombus muscorum (Linnaeus, 1758).

We present a genome assembly from an individual female Bombus muscorum (the moss carder bee; Arthropoda; Insecta; Hymenoptera; Apidae). The genome sequence spans 317.70 megabases. Most of the assembly is scaffolded into 17 chromosomal pseudomolecules. The mitochondrial genome has also been assembled and is 21.15 kilobases in length. Gene annotation of this assembly on Ensembl identified 11,668 protein-coding genes.

The genome sequence of a braconid wasp, Aleiodes testaceus (Telenga, 1941).

We present a genome assembly from an individual female Aleiodes testaceus (braconid wasp; Arthropoda; Insecta; Hymenoptera; Braconidae). The genome sequence spans 110.70 megabases. Most of the assembly is scaffolded into 19 chromosomal pseudomolecules. The mitochondrial genome has also been assembled and is 28.0 kilobases in length. Gene annotation of this assembly on Ensembl identified 10,520 protein-coding genes.

The genome sequence of an ichneumonid wasp, Netelia fuscicornis (Holmgren, 1860).

We present a genome assembly from an individual female Netelia fuscicornis (an ichneumonid wasp; Arthropoda; Insecta; Hymenoptera; Ichneumonidae). The genome sequence is 324.7 megabases in span. Most of the assembly is scaffolded into six chromosomal pseudomolecules. The mitochondrial genome has also been assembled and is 17.7 kilobases in length.

The genome sequence of an ichneumonid wasp, Cylloceria caligata (Gravenhorst, 1829).

We present a genome assembly from an individual male Cylloceria caligata (an ichneumonid wasp; Arthropoda; Insecta; Hymenoptera; Ichneumonidae). The genome sequence spans 596.20 megabases. Most of the assembly is scaffolded into 9 chromosomal pseudomolecules. The mitochondrial genome has also been assembled and is 33.21 kilobases in length.

The genome sequence of an ichneumonid wasp, Rhimphoctona ( Xylophylax) megacephalus (Gravenhorst, 1829).

We present a genome assembly from an individual female Rhimphoctona megacephalus (ichneumonid wasp; Arthropoda; Insecta; Hymenoptera; Ichneumonidae). The genome sequence spans 406.00 megabases. Most of the assembly is scaffolded into 11 chromosomal pseudomolecules. The mitochondrial genome has also been assembled and is 28.53 kilobases in length.

The genome sequence of a green lacewing, Nineta flava (Scopoli, 1763).

We present a genome assembly from an individual Nineta flava (green lacewing; Arthropoda; Insecta; Neuroptera; Chrysopidae). The genome sequence spans 732.30 megabases. Most of the assembly is scaffolded into 7 chromosomal pseudomolecules. The mitochondrial genome has also been assembled and is 16.16 kilobases in length.

The genome sequence of an ichneumonid wasp, Oxytorus armatus Thomson, 1883.

We present a genome assembly from an individual male Oxytorus armatus (an ichneumonid wasp; Arthropoda; Insecta; Hymenoptera; Ichneumonidae). The genome sequence is 367.8 megabases in span. Most of the assembly is scaffolded into 13 chromosomal pseudomolecules. The mitochondrial genome has also been assembled and is 56.22 kilobases in length.

New Species of Grotea (Hymenoptera, Ichneumonidae, Labeninae) from Ecuador, with New Records and a Key to the Neotropical Species.

Here we describe two new Grotea species from Ecuador, G. akakana Mazón & Bordera sp. nov., and G. romeri Mazón sp. nov., as well as the male of G. cundinamarquesa Herrera-Flórez 2018. G. akakana sp. nov. is characterized by the combination of a postgenal process long, a 45-flagellomeres antenna without a white band and a mesopleuron black with two yellow spots separated by a red one. On the other hand, G. romeri sp. nov. is characterized by the combination of a postgenal process very short, a 36-flagellomeres antenna without a white band, a propodeum with a long and narrow area lateralis, uninterrupted yellow-colored orbits and a mesopleuron black with a yellow spot in the middle. The species G. santandereana Herrera-Flórez 2018 and G. surinamese Herrera-Flórez 2019 are recorded from Ecuador for the first time. This brings the total of described Grotea species to 31, all from the New World, with 27 of these exclusively Neotropical. A key for the identification of Neotropical species is included.

The genome sequence of the chalcid wasp, Chalcis sispes Linnaeus, 1761.

We present a genome assembly from an individual female Chalcis sispes (chalcid wasp; Arthropoda; Insecta; Hymenoptera; Chalcididae). The genome sequence is 412.4 megabases in span. Most of the assembly is scaffolded into 6 chromosomal pseudomolecules. The mitochondrial genome has also been assembled and is 15.9 kilobases in length.