The de novo genome of the Black-necked Snakefly (Venustoraphidia nigricollis Albarda, 1891): A resource to study the evolution of living fossils.

Snakeflies (Raphidioptera) are the smallest order of holometabolous insects that have kept their distinct and name-giving appearance since the Mesozoic, probably since the Jurassic, and possibly even since their emergence in the Carboniferous, more than 300 million years ago. Despite their interesting nature and numerous publications on their morphology, taxonomy, systematics, and biogeography, snakeflies have never received much attention from the general public, and only a few studies were devoted to their molecular biology. Due to this lack of molecular data, it is therefore unknown, if the conserved morphological nature of these living fossils translates to conserved genomic structures. Here, we present the first genome of the species and of the entire order of Raphidioptera. The final genome assembly has a total length of 669 Mbp and reached a high continuity with an N50 of 5.07 Mbp. Further quality controls also indicate a high completeness and no meaningful contamination. The newly generated data was used in a large-scaled phylogenetic analysis of snakeflies using shared orthologous sequences. Quartet score and gene concordance analyses revealed high amounts of conflicting signals within this group that might speak for substantial incomplete lineage sorting and introgression after their presumed re-radiation after the asteroid impact 66 million years ago. Overall, this reference genome will be a door-opening dataset for many future research applications, and we demonstrated its utility in a phylogenetic analysis that provides new insights into the evolution of this group of living fossils.

Unraveling the evolutionary history of the snakefly family Inocelliidae (Insecta: Raphidioptera) through integrative phylogenetics.

Inocelliidae is one of the two extant families of the holometabolan order Raphidioptera (snakeflies), with the modern fauna represented by seven genera and 44 species. The evolutionary history of the family is little-known. Here we present the first phylogenetic and biogeographical analyses based on a worldwide sampling of taxa and datasets combined with morphological characters and mitochondrial genomes, aiming to investigate the intergeneric phylogeny and historical biogeography of Inocelliidae. The phylogenetic inference from the combined analysis of morphological and molecular data recovered the sister-group relationship between a clade of (Negha + Indianoinocellia) + Sininocellia and a clade of Fibla + the Inocellia clade (interiorly nested by Amurinocellia and Parainocellia). Amurinocellia stat.r. and Parainocellia stat.r. et emend.n. are relegated to subgeneric status within Inocellia, whereas a newly erected subgenus of Inocellia, Epinocellia subgen.n., accommodates the former Parainocellia burmana (U. Aspöck and H. Aspöck, 1968) plus a new species Inocellia (Epinocellia) weii sp.n. Further, the Inocellia crassicornis group constitutes the nominotypical subgenus Inocellia stat.n., but the Inocellia fulvostigmata group is paraphyletic. Diversification within Inocelliidae is distinguished by an Eocene divergence leading to extant genera and a Miocene radiation of species. A biogeographical scenario depicts how the diverse inocelliid fauna from East Asia could have originated from western North America via dispersal across the Beringia during the early Tertiary, and how the Miocene ancestors of Inocellia could have accomplished long-distance dispersals via the Tibet-Himalayan corridor or eastern Palaearctic to western Palaearctic. Our results shed new light specifically on the evolution of Inocelliidae and, in general, the Raphidioptera.

Similar pattern, different paths: tracing the biogeographical history of Megaloptera (Insecta: Neuropterida) using mitochondrial phylogenomics.

The sequential breakup of the supercontinent Pangaea since the Middle Jurassic is one of the crucial factors that has driven the biogeographical patterns of terrestrial biotas. Despite decades of effort searching for concordant patterns between diversification and continental fragmentation among taxonomic groups, increasing evidence has revealed more complex and idiosyncratic scenarios resulting from a mixture of vicariance, dispersal and extinction. Aquatic insects with discreet ecological requirements, low vagility and disjunct distributions represent a valuable model for testing biogeographical hypotheses by reconstructing their distribution patterns and temporal divergences. Insects of the order Megaloptera have exclusively aquatic larvae, their adults have low vagility, and the group has a highly disjunct geographical distribution. Here we present a comprehensive phylogeny of Megaloptera based on a large-scale mitochondrial genome sequencing of 99 species representing >90% of the world genera from all major biogeographical regions. Molecular dating suggests that the deep divergence within Megaloptera pre-dates the breakup of Pangaea. Subsequently, the intergeneric divergences within Corydalinae (dobsonflies), Chauliodinae (fishflies) and Sialidae (alderflies) might have been driven by both vicariance and dispersal correlated with the shifting continent during the Cretaceous, but with strikingly different and incongruent biogeographical signals. The austral distribution of many corydalids appears to be a result of colonization from Eurasia through southward dispersal across Europe and Africa during the Cretaceous, whereas a nearly contemporaneous dispersal via northward rafting of Gondwanan landmasses may account for the colonization of extant Eurasian alderflies from the south.

Mining the Species Diversity of Lacewings: New Species of the Pleasing Lacewing Genus Dilar Rambur, 1838 (Neuroptera, Dilaridae) from the Oriental Region.

The species diversity of insects is extraordinarily rich, but still has been insufficiently explored or underestimated particularly for uncommon groups. The pleasing lacewings (Dilaridae) are a little known family of Neuroptera with distinct sexually dimorphic antennae. The species diversity of pleasing lacewings was recently found to be severely underestimated and requires a comprehensive investigation, as well as systematic reviews. Here, we report on 12 new species of the pleasing lacewing genus Dilar Rambur, 1838, from the Oriental region, namely D. forcipatus sp. nov. and D. laoticus sp. nov. from Laos (new country record of Dilar); D. malickyi sp. nov., D. phraenus sp. nov. and D. rauschorum sp. nov. from northern Thailand; D. striatus sp. nov. from northern Vietnam; D. cangyuanensis sp. nov., D. daweishanensis sp. nov., D. nujianganus sp. nov., D. weibaoshanensis sp. nov., D. yucheni sp. nov., and D. zhangweiae sp. nov. from Yunnan and Tibet, both in southwestern China. The new species of Dilar display several types of wing marking patterns, and the morphology of the male genitalia is highly diverse. A comprehensive examination of the species diversity and distribution of Dilar concluded that Yunnan (southwestern China) represents a biogeographic region with high endemism and the richest species diversity. The potential correlation between vertical distribution and geographical latitude in Dilar was also analyzed.

Comparative morphology of extant raptorial Mantispoidea (Neuroptera: Mantispidae, Rhachiberothidae) suggests a non-monophyletic Mantispidae and a single origin of the raptorial condition within the superfamily.

Adult external morphology of the extant raptorial Mantispoidea (Insecta: Neuroptera: Mantispidae and Rhachiberothidae) is compared emphasizing the morphology of the subfamily Symphrasinae as a key group to understand the phylogenetic relationships among the members of the superfamily. Plega dactylota Rehn, 1939 is thoroughly characterized in order to exemplify the morphology of the Symphrasinae. Additionally, following a review of the literature and examination of comparative material of Dilaridae, Berothidae, Rhachiberothidae and all Mantispidae subfamilies, a new interpretation of the components of the raptorial apparatus (i.e., head, prothorax, grasping forelegs, as well as integumentary specializations) is presented. Also, wing venation for these groups is reinterpreted, and new homology hypotheses for wing venation are proposed based on tracheation and comparative analyses. Given the high morphological divergence on the genital sclerites within the Mantispoidea, plus the confusing previous usage of neutral terminology and terms referring to appendages across taxonomic and morphological studies, we attempt to standardize, simplify, and situate terminology in an evolutionary context under the "gonocoxite concept" (multi-coxopod hypothesis). The remarkable morphological similarity of the genital sclerites of Symphrasinae and Rhachiberothidae (sensu U. Aspöck Mansell 1994) with the Nallachinae (Dilaridae) was taken as a starting point to understand the morphology of other Mantispidae subfamilies. Based on these morphological comparisons, we provide a revised phylogenetic analysis of Mantispoidea. This new phylogenetic analysis supports a sister group relationship between the family Rhachiberothidae, comprising Rhachiberothinae and Symphrasinae, and the family Mantispidae, including the subfamily Mantispinae and its sister taxa Drepanicinae and Calomantispinae, which may represent a single subfamily. Based on these analyses, raptorial condition probably evolved a single time in these insects and subsequently became diversified in the two sister clades of the raptorial Mantispoidea.

New beaded lacewings (Insecta: Neuroptera: Berothidae) from Indochina.

The family Berothidae (beaded lacewings) is poorly known from the Indochina Peninsula. Previously, three genera, Berotha Walker, 1860, Isoscelipteron Costa, 1863, and Lekrugeria Navás, 1929, and four species were recorded from Myanmar, Thailand, and Vietnam. Here, we record the genus Asadeteva U. Aspöck H. Aspöck, 1981, in Indochina for the first time and provide a description of a new species, namely Asadeteva acutata sp. nov., from Laos and Thailand. We also describe a new species of the genus Berotha Walker, 1860, namely Berotha incurvata sp. nov. from Laos and provide the first description of the male of Lekrugeria nepalica U. Aspöck H. Aspöck, 1986, based on material from northern Myanmar.

Correction to: An integrative phylogenomic approach to elucidate the evolutionary history and divergence times of Neuropterida (Insecta: Holometabola).

An amendment to this paper has been published and can be accessed via the original article.

Rhachiella malawica gen. nov., spec. nov. from Malawi-another beauty of the Afrotropics (Neuroptera: Rhachiberothidae).

A new species and a new genus of Rhachiberothidae, Rhachiella malawica gen. nov., spec. nov., are described from Malawi. The new species is characterized by a flat vertex, a long penisfilum in the male, and by a bifurcate pseudohypocauda in the female. This combination of characters requires the description of a new genus, which is the sister taxon of Mucroberotha Tjeder, 1959. This is the first record of Rhachiberothidae in Malawi. The distributions of all 14 species of Rhachiberothidae so far known are shown in three maps.

An integrative phylogenomic approach to elucidate the evolutionary history and divergence times of Neuropterida (Insecta: Holometabola).

BACKGROUND: The latest advancements in DNA sequencing technologies have facilitated the resolution of the phylogeny of insects, yet parts of the tree of Holometabola remain unresolved. The phylogeny of Neuropterida has been extensively studied, but no strong consensus exists concerning the phylogenetic relationships within the order Neuroptera. Here, we assembled a novel transcriptomic dataset to address previously unresolved issues in the phylogeny of Neuropterida and to infer divergence times within the group. We tested the robustness of our phylogenetic estimates by comparing summary coalescent and concatenation-based phylogenetic approaches and by employing different quartet-based measures of phylogenomic incongruence, combined with data permutations. RESULTS: Our results suggest that the order Raphidioptera is sister to Neuroptera + Megaloptera. Coniopterygidae is inferred as sister to all remaining neuropteran families suggesting that larval cryptonephry could be a ground plan feature of Neuroptera. A clade that includes Nevrorthidae, Osmylidae, and Sisyridae (i.e. Osmyloidea) is inferred as sister to all other Neuroptera except Coniopterygidae, and Dilaridae is placed as sister to all remaining neuropteran families. Ithonidae is inferred as the sister group of monophyletic Myrmeleontiformia. The phylogenetic affinities of Chrysopidae and Hemerobiidae were dependent on the data type analyzed, and quartet-based analyses showed only weak support for the placement of Hemerobiidae as sister to Ithonidae + Myrmeleontiformia. Our molecular dating analyses suggest that most families of Neuropterida started to diversify in the Jurassic and our ancestral character state reconstructions suggest a primarily terrestrial environment of the larvae of Neuropterida and Neuroptera. CONCLUSION: Our extensive phylogenomic analyses consolidate several key aspects in the backbone phylogeny of Neuropterida, such as the basal placement of Coniopterygidae within Neuroptera and the monophyly of Osmyloidea. Furthermore, they provide new insights into the timing of diversification of Neuropterida. Despite the vast amount of analyzed molecular data, we found that certain nodes in the tree of Neuroptera are not robustly resolved. Therefore, we emphasize the importance of integrating the results of morphological analyses with those of sequence-based phylogenomics. We also suggest that comparative analyses of genomic meta-characters should be incorporated into future phylogenomic studies of Neuropterida.

First description of male genital sclerites and associated musculature for two members of Coniopterygidae (Insecta: Neuropterida: Neuroptera) based on X-ray microCT imaging.

Coniopterygidae are the dwarfs among the Neuroptera. Despite their miniaturisation, the males are equipped with genital sclerites that are excessively heterogeneous. They function in copulation and sperm transfer and have been widely utilized for species identification, as well being considered of high phylogenetic relevance. The present study is the first to describe the musculature associated with the genital sclerites of two species of Coniopterygidae, Helicoconis lutea (Wallengren, 1871) (Aleuropteryginae), and Coniopteryx pygmaea (Enderlein, 1906) (Coniopteryginae) based on X-ray microCT imaging. We found six pairs of muscles associated with the genital sclerites in H. lutea and seven in C. pygmaea. The images depict other internal organs of the posterior abdominal segments, such as gonads and alimentary canal. In both investigated species, the internal sclerites support the ductus ejaculatorius, which - surprisingly - turned out to be a landmark for the identification of closely adjacent internal sclerites and associated musculature. The interpretation of these sclerites as gonocoxites and gonapophyses of the tenth segment (traditionally denoted as parameres and penis) could be corroborated. Thus it is no longer tenable to assert that possession of a "penis" is exclusive to Coniopterygidae, since this sclerite is part of the ground pattern in Neuroptera. Interactions of genital sclerites and corresponding musculature during copulation are discussed.

The giant "penis" of a newly described species in the subgenus Scotoconiopteryx from Peru (Neuroptera: Coniopterygidae: Coniopteryginae: Coniopteryx).

The coniopterygid fauna of the Neotropical region is poorly known. The most recent summary was provided by Martins (2019) who noted that 14 species were known from Peru. The subgenus Coniopteryx (Scotoconiopteryx) is endemic to the Neotropical region and comprises 36 species (Meinander 1972; Meinander Penny 1982; Sziráki 2011; Martins Amorim 2016, Sarmiento-Cordero Contreras-Ramos 2019).

A review of the pleasing lacewing genus Dilar Rambur (Neuroptera, Dilaridae) from South Asia.

The pleasing lacewing genus Dilar Rambur, 1838, is poorly known from South Asia, previously only three species were accounted. Here, we present a revision of Dilar from South Asia, with descriptions of five new species, namely Dilar austroindicus sp. nov., Dilar biprojectus sp. nov., Dilar clavatus sp. nov., Dilar miralobatus sp. nov., and Dilar truncatus sp. nov. In addition, Dilar nietneri Hagen, 1858, from Sri Lanka is re-described. A key to males of Dilar from South Asia is provided.

A review of the pleasing lacewing genus Dilar Rambur (Neuroptera, Dilaridae) from Central Asia.

The pleasing lacewing genus Dilar Rambur, 1838, is the only known representative of the family Dilaridae from Central Asia. Here we present a review of the five species of Dilar from Central Asia, Dilar hornei McLachlan, 1869, Dilar caesarulus H. Aspöck U. Aspöck, 1967, Dilar kirgisus H. Aspöck U. Aspöck, 1967, Dilar vartianorum H. Aspöck U. Aspöck, 1967, and Dilar dochaner H. Aspöck U. Aspöck, 1968. Remarkable intraspecific variations in male gonocoxite 10 are found in D. kirgisus and D. vartianorum. Re-descriptions of all the species are provided. Dilar indicus Monserrat, 1989, and Dilar similis Monserrat, 1989, are synonymized with D. vartianorum. A key to the species of Dilar from Central Asia is provided.

A review of the beaded lacewings (Neuroptera: Berothidae) from China.

The lacewing family Berothidae (beaded lacewings) of China is a poorly known group. It consists of two genera and 11 previously described species. In this paper, we provide a revision of the beaded lacewings from China. The genus Berotha Walker, 1860, in China contains seven species including one new species: Berotha guangdongana sp. nov. The second genus, Isoscelipteron Costa, 1863, comprises six species from China, including one new species, namely Isoscelipteron acuticaudatum sp. nov. Descriptions and re-descriptions of the species are provided as well as an identification key to the species of Berothidae from China.

Discovery of a new species of Inocelliidae (Insecta: Raphidioptera) in an altitude of nearly 3500 m in China.

A new snakefly species of the family Inocelliidae is described from China: Inocellia occidentalis sp. nov. The new species belongs to the Inocellia crassicornis species group.

Taxonomic notes on Cretarophalis patrickmuelleri Wichard, 2017 (Insecta: Neuroptera: Nevrorthidae) from the mid-Cretaceous of Myanmar, and its phylogenetic significance.

Nevrorthidae is a family of Neuroptera with low species diversity, disjunct geographic distributions, and a controversial phylogenetic status. Previous fossil records of the family are from the Eocene except for the recently described species Cretarophalis patrickmuelleri Wichard, 2017 from mid-Cretaceous Burmese amber, following earlier records of an undescribed species. However, such a significant finding of this family from Mesozoic was originally presented only with a preliminary description. Here we re-describe the species based on exquisitely preserved materials from mid-Cretaceous Burmese amber that provides for a much more detailed description, in particular of the hindwing, female genitalia, and larval head. Furthermore, distinctive morphological characters of C. patrickmuelleri with significant phylogenetic relevance are discussed. The general morphology, particularly that of the larva, has changed little between the Mesozoic and Cenozoic, and it reflects an evolutionary stasis that might correspond to their aquatic larval life-style. The present finding also supports the relictual nature of the modern nevrorthids.

New species of the snakefly genus Mongoloraphidia (Raphidioptera: Raphidiidae) from China.

Two new snakefly species of the family Raphidiidae are described from China: Mongoloraphidia lini sp. nov. and Mongoloraphidia triangulata sp. nov. Biogeographical considerations on secondary refugial centers of the large Sino-Tibetan primary center are presented.

Mitochondrial phylogenomics illuminates the evolutionary history of Neuropterida.

Neuroptera (lacewings) and allied orders Megaloptera (dobsonflies, alderflies) and Raphidioptera (snakeflies) are predatory insects and together make up the clade Neuropterida. The higher-level relationships within Neuropterida have historically been widely disputed with multiple competing hypotheses. Moreover, the evolution of important biological innovations among various Neuropterida families, such as the origin, timing and direction of transitions between aquatic and terrestrial habitats of larvae, remains poorly understood. To investigate the origin and diversification of lacewings and their allies, we undertook phylogenetic analyses of mitochondrial genomes of all families of Neuropterida using Bayesian inference, maximum likelihood and maximum parsimony methods. We present a robust, fully resolved phylogeny and divergence time estimation for Neuropterida with strong statistical support for almost all nodes. Mitochondrial sequence data are typified by significant compositional heterogeneity across lineages, and parsimony and models assuming homogeneous rates did not recover Neuroptera as monophyletic. Only a model accounting for compositional heterogeneity (i.e. CAT-GTR) recovered all orders of Neuropterida as monophyletic. Significant findings of the mitogenomic phylogeny include recovering Raphidioptera as sister to Megaloptera plus Neuroptera. The sister family of all other lacewings are the dusty-wings (Coniopterygidae), rather than Nevrorthidae. Nevrorthidae are instead returned to their traditional position as the sister group of the spongilla-flies (Sisyridae) and closely related to Osmylidae. Our divergence time analysis indicates that the Mesozoic was indeed a 'golden age' for lacewings, with most families of Neuropterida diverging during the Triassic and Jurassic and all extant families present by the Early Cretaceous. Based on ancestral character state reconstructions of larval habitat we evaluate competing hypotheses regarding the life style of early neuropteridan larvae as either aquatic or terrestrial.

Head anatomy of adult Coniopteryx pygmaea: Effects of miniaturization and the systematic position of Coniopterygidae (Insecta: Neuroptera).

External and internal head structures of adult Coniopteryx pygmaea Enderlein, 1906, one of the smallest known lacewings, are described in detail for the first time. Possible effects of miniaturization and two hypotheses on the phylogenetic position of Coniopterygidae are evaluated and compared with data from literature. Several convergent modifications in C. pygmaea and other miniaturized insect species are outlined, e.g., a relative increase in the size of the brain, simplification of the tracheal system with respect to the number of tracheae, and reduction of the number of ommatidia and diameter of the facets. Further, the ocular ridge is bell-shaped and countersunk into the head capsule. The cuticle is weakly sclerotized and equipped with wax glands which are unique in Neuroptera. The total number of muscles is not affected by miniaturization. The phylogenetic analysis yields Coniopterygidae as sistergroup to the dilarid clade based on one larval character, the shape of the stylets. The enforced basal position of Coniopterygidae is supported by one disputable synapomorphy of the remaining Neuroptera, the presence of paraglossae in adults.