Abstract Butterflies have been of great interest to naturalists for centuries, and the study of butterflies has been an integral part of ecology and evolution ever since Darwin proposed his theory of natural selection in 1859. There are > 18 000 butterfly species worldwide, showing great diversity in morphological traits and ecological niches. Compared with butterfly diversity, however, patterns of genome size variation in butterflies remain poorly understood, especially in a phylogenetic context. Here, we sequenced and assembled the mitogenomes of 68 butterflies and measured the genome sizes ( C ‐values) of 67 of them. We also assembled 10 mitogenomes using reads from GenBank. Among the assembled 78 mitogenomes, those from 59 species, 23 genera and one subfamily are reported for the first time. Combining with published data of mitogenomes and genome size, we explored the patterns in genome size variation for 106 butterfly species in a phylogenetic context based on analyses of mitogenomes from 264 species covering six families. Our results show that the genome size of butterflies has a 6.4‐fold variation ranging from 0.203 pg (199 Mb) (Nymphalidae: Heliconius xanthocles ) to 1.287 pg (1253 Mb) (Papilionidae: Parnassius orleans ). Within families, the largest variation was found in Papilionidae (5.9‐fold: 0.22–1.29 pg), followed by Nymphalidae (4.8‐fold: 0.2–0.95 pg), Pieridae (4.4‐fold: 0.22–0.97 pg), Hesperiidae (2.2‐fold: 0.3–0.66 pg), Lycaenidae (2.6‐fold: 0.39–1.02 pg) and Rioidinidae (1.8‐fold: 0.48–0.87 pg). Our data also suggest that butterflies have an ancestral genome size of c. 0.5 pg, and some ancestral genome size increase or decrease events along different subfamilies or tribes produce the diversity of genome size variation in diverse butterflies. Our data provide novel insights into patterns of genome size variation in butterflies and are an important reference for future genome sequencing programmes.

Abstract For regulatory purposes, the name ‘Asian gypsy moth’ refers to a group of closely related Asian Lymantria species and subspecies whose female moths display flight capability, a trait believed to confer enhanced invasiveness relative to the European gypsy moth, Lymantria dispar dispar , whose females are flightless. Lymantria albescens and Lymantria postalba are Asian gypsy moths occurring in the southern Ryukyu Islands and in the northern Ryukyu and adjacent Kyushu and Shikoku Islands of Japan, respectively. Although once considered subspecies of L. dispar , their status as distinct species, relative to the latter, is now well established. While postalba was subsequently considered a subspecies of L. albescens , largely on the basis of differences in forewing ground colour in males, both taxa were later given distinct species status by Pogue & Schaefer (2007) following their revision of the genus Lymantria . Here, we re‐examined the validity of this revised status through the sequencing of a large portion of the mitochondrial genome ( c . 60%) and multiple nuclear marker genes [elongation factor 1‐alpha (Ef‐1α), wingless (Wgl), internal transcribed spacer 2 (ITS‐2), ribosomal protein S5 (RpS5)] in representative specimens of both taxa and other Lymantria species, including L. monacha , L. xylina , L. mathura and members of the L. dispar + L. umbrosa clade. A comparison of the number of substitutions in these genomic regions among the taxa we considered showed lower or equivalent variation between L. albescens and L. postalba compared with subspecies of L. dispar , for mitochondrial and nuclear sequences, respectively. This finding was reflected in the maximum likelihood trees generated independently for mitochondrial and nuclear data, where L. albescens and L. postalba formed, in both analyses, a short‐branch sister clade basal to the L. dispar + L. umbrosa clade. We further sequenced three markers [cytochrome c oxydase 1 (COI), EF‐1α, Wgl] in multiple L. albescens – L. postalba specimens collected along a south‐to‐north transect across the Ryukyu Arc and observed no clear distinction among the sampled specimens as a function of taxonomic designation. We conclude that L. albescens and L. postalba form a single species, with postalba representing a darker‐winged morph along an apparent south‐to‐north wing colour cline. Accordingly, L. postalba is relegated to synonymy under L. albescens ( syn.n. ).

Abstract Zoraptera is one of the most enigmatic and least understood orders in insects. Based on a wide taxon sampling from all continents where the group is known, we applied a phylogenetic approach using multiple DNA sequences to elucidate species‐level relationships. The resulting phylogeny shows that Zoraptera is divided into three major clades, and that two comprise species distributed on different continents. The monophyly of these clades is at least partly supported by shared derived morphological features. The divergence age estimation and ancestral distribution area reconstruction suggest an ancient origin and early radiation initiated in the Permian. Plate tectonics theory suggests that the present distribution of Zoraptera was mainly established by vicariance, rather than dispersal. The three major clades probably originated on the Pangaea supercontinent, or alternatively on the linked Gondwana and Laurasia supercontinents. Their ancient origin explains previously found conspicuous interspecific variation of the genital apparatus, sperm structure and mating behaviour, in striking contrast to a highly conserved general body morphology. We compiled data of available reproductive features and reconstructed the character evolution. Our analyses revealed repeated acquisitions and/or losses of a hyperelongated intromittent organ, mating hooks and tergal protuberances.

Abstract The phylogenetic interrelationships among four hexapod lineages (Protura, Collembola, Diplura and Insecta) are pivotal to understanding the origin of insects and the early diversification of Hexapoda, but they have been difficult to clarify based on the available data. In this study, we identified 91 conserved microRNA (miRNA) families from 36 panarthropod taxa, including seven newly sequenced non‐insect hexapods. We found major clade differentiation accompanied by the origin of novel miRNA families, and most miRNA clusters are conserved with a high degree of microsynteny. Importantly, we were able to identify two miRNA families unique to Hexapoda, and four miRNA families and a miRNA cluster that exist exclusively in Diplura and Insecta, suggesting a close relationship between Diplura and Insecta as well as the monophyly of Hexapoda. Combined with a phylogenetic analysis based on the presence/absence matrix of miRNA families, our study demonstrates the effectiveness of miRNA in resolving deep phylogenetic problems.

Abstract Triassic fossils are rare but crucial for understanding the early evolution of large insect clades including beetles (Coleoptera). Their phylogenetic assignment is problematic because of fragmentary preservation, yet crucial for the correct use of the information they provide. Here an analysis is presented of the phylogenetic position of Leehermania prorova , the Late Triassic compressed fossil which was described and hitherto widely used as the oldest representative of Staphylinidae (rove beetles) in the suborder Polyphaga. By contrast with the intuitive character assessment made in the original description, a phylogenetic analysis of Leehermania is performed using an extensive morphological matrix of extant Coleoptera provided by the Beetle Tree of Life project, constrained in view of the latest relevant molecular phylogenies. As a result, Leehermania is identified as an extinct lineage within the beetle suborder Myxophaga, closest to the modern family Hydroscaphidae. Excluding Leehermania from Staphylinidae and placing it in Myxophaga amends erroneous assumptions about early diversification of rove beetles and enhances our views of the evolutionary history of Coleoptera.

Abstract The high‐level classification of Chrysomelidae (leaf beetles) currently recognizes 12 or 13 well‐established subfamilies, but the phylogenetic relationships among them remain ambiguous. Full mitochondrial genomes were newly generated for 27 taxa and combined with existing GenBank data to provide a dataset of 108 mitochondrial genomes covering all subfamilies. Phylogenetic analysis under maximum likelihood and Bayesian inference recovered the monophyly of all subfamilies, except that Timarcha was split from Chrysomelinae in some analyses. Three previously recognized major clades of Chrysomelidae were broadly supported: the ‘chrysomeline’ clade consisting of (Chrysomelinae (Galerucinae + Alticinae)); the ‘sagrine’ clade with internal relationships of ((Bruchinae + Sagrinae) + (Criocerinae + Donaciinae)), and the ‘eumolpine’ clade comprising (Spilopyrinae (Cassidinae (Eumolpinae (Cryptocephalinae + Lamprosomatinae)))). Relationships among these clades differed between data treatments and phylogenetic algorithms, and were complicated by two additional deep lineages, Timarcha and Synetinae. Various topological tests favoured the PhyloBayes software as the preferred inference method, resulting in the arrangement of (chrysomelines (eumolpines + sagrines)), with Timarcha placed as sister to the chrysomeline clade and Synetinae as a deep lineage splitting near the base. Whereas mitogenomes provide a solid framework for the phylogeny of Chrysomelidae, the basal relationships do not agree with the topology of existing molecular studies and remain one of the most difficult problems of Chrysomelidae phylogenetics.

Abstract The hemipteran suborder Auchenorrhyncha is a highly diverse, ecologically and agriculturally important group of primarily phytophagous insects which has been a source of phylogenetic contention for many years. Here, we have used transcriptome sequencing to assemble 2139 orthologues from 84 auchenorrhynchan species representing 27 families; this is the largest and most taxonomically comprehensive phylogenetic dataset for this group to date. We used both maximum likelihood and multispecies coalescent analyses to reconstruct the evolutionary history in this group using amino acid, nucleotide, and degeneracy‐coded nucleotide orthologue data. Although many relationships at the superfamily level were consistent between analyses, several differing, highly supported topologies were recovered using different datasets and reconstruction methods, most notably the differential placement of Cercopoidea as sister to either Cicadoidea or Membracoidea. To further interrogate the recovered topologies, we explored the contribution of genes as partitioned by third‐codon‐position guanine‐cytosine (GC) content and heterogeneity. We found consistent support for several relationships, including Cercopoidea + Cicadoidea, most often in genes that would be expected to be enriched for the true species tree if recombination‐based dynamics in GC content have contributed to the observed GC heterogeneity. Our results provide a generally well‐supported framework for future studies of auchenorrhynchan phylogeny and suggest that transcriptome sequencing is likely to be a fruitful source of phylogenetic data for resolving its clades. However, we caution that future work should account for the potential effects of GC content heterogeneity on relationships recovered in this group.

Abstract In this study we analysed for the first time the phylogenetic relationships among the European Lethrus species based on mitochondrial (COI, 16S) and nuclear (H3, wingless) markers. The phylogenetic inference based on these genes indicated that the European Lethrus species form a monophyletic group with two main lineages. One of them is distributed in the central part of the Balkan Peninsula (North Macedonia, northwestern Greece and southwestern Bulgaria) and includes five species ( L. perun , L. raymondi , L. strymonensis , L. halkidikiensis and L. elephas ). Three species of the other clade ( L. schaumii , L. ares and L. schneideri ) occur in the eastern part of the Balkan Peninsula (southeastern Bulgaria and northeastern Greece) while its fourth member, L. apterus , could colonize much larger areas in central and eastern Europe. Despite its large geographic distribution, L. apterus proved to be a single species. By contrast, we found strong intraspecific differentiation in both L. schaumii and L. halkidikiensis in spite of their restricted distribution. Surprisingly, two highly divergent mitogroups were also detected within a single L. schneideri population. As the microsatellite markers did not show any differentiation between them, this finding is likely to be accounted for by a unidirectional hybridization event. These findings highlight that phylogenetic studies or species descriptions should not be based on mitochondrial markers alone. Our study clearly demonstrates that allopatric speciation is the most important speciation mechanism in the European species of the genus Lethrus , although we also hypothesize that other well‐known factors, such as hybridization or the glacial‐interglacial dynamics, could significantly affect the speciation patterns of certain lineages.