The Complete Mitochondrial Genome and Phylogenetic Analysis of Rhagastis binoculata (Matsumura, 1909) (Lepidoptera: Sphingidae).

The mitochondrial genome (mitogenome) Rhagastis binoculata (Matsumura, 1909), an endemic moth species in Taiwan, was sequenced and analyzed. The complete circular mitogenome of R. binoculata is 15,303 bp and contains 13 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes, and an AT-rich control region. The mitogenome has an overall nucleotide composition of 41.2% A, 11.9% C, 7.5% G, and 39.4% T, with an AT content of 80.6%. Of the protein-coding genes (PCGs), 12 start with ATG, ATT, and ATC, and COX1 starts with a "CGA" codon. All of the stop codons are "TAA, TAG, or T". Our phylogenetic analysis of 21 species of Sphingidae insects suggests that R. binoculata is clustered with Rhagastis mongoliana, which belongs to the subfamily Macroglossinae.

Phylogenetic structure of moth communities (Geometridae, Lepidoptera) along a complete rainforest elevational gradient in Papua New Guinea.

We use community phylogenetics to elucidate the community assembly mechanisms for Geometridae moths (Lepidoptera) collected along a complete rainforest elevational gradient (200-3700 m a.s.l) on Mount Wilhelm in Papua New Guinea. A constrained phylogeny based on COI barcodes for 604 species was used to analyse 1390 species x elevation occurrences at eight elevational sites separated by 500 m elevation increments. We obtained Nearest Relatedness Index (NRI), Nearest Taxon Index (NTI) and Standardised Effect Size of Faith's Phylogenetic Diversity (SES.PD) and regressed these on temperature, plant species richness and predator abundance as key abiotic and biotic predictors. We also quantified beta diversity in the moth communities between elevations using the Phylogenetic Sorensen index. Overall, geometrid communities exhibited phylogenetic clustering, suggesting environmental filters, particularly at higher elevations at and above 2200 m a.s.l and no evidence of overdispersion. NRI, NTI and SES.PD showed no consistent trends with elevation or the studied biotic and abiotic variables. Change in community structure was driven by turnover of phylogenetic beta-diversity, except for the highest 2700-3200 m elevations, which were characterised by nested subsets of lower elevation communities. Overall, the elevational signal of geometrid phylogeny was weak-moderate. Additional insect community phylogeny studies are needed to understand this pattern.

Phylogenetic relationships and historical biogeography of silkmoths (Lepidoptera: Bombycidae) suggest an origin in Southern Gondwana.

Silkmoths (Bombycidae) have a disjunct distribution predominantly in the Southern Hemisphere and Asia. Here we reconstruct the phylogenetic history of the family to test competing hypotheses on their origin and assess how vicariance and long-distance dispersal shaped their current distribution. We sequenced up to 5,074 base pairs from six loci (COI, EF1-α, wgl, CAD, GAPDH, and RpS5) to infer the historical biogeography of Bombycidae. The multilocus dataset covering 20 genera (80 %) of the family, including 17 genera (94 %) of Bombycinae and 3 genera (43 %) of Epiinae, was used to estimate phylogenetic patterns, divergence times and biogeographic reconstruction. Dating estimates extrapolated from secondary calibration sources indicate the Bombycidae stem-group originated approximately 64 Mya. The subfamilies Epiinae (South America) and Bombycinae (Australia, Asia, East Palaearctic, and Africa) were reciprocally monophyletic, diverging at c. 56 Mya (95 % credibility interval: 66-46 Mya). The 'basal' lineage of Bombycinae - Gastridiota + Elachyophtalma - split from the rest of Bombycinae c. 53 Mya (95 % credibility interval: 63-43 Mya). Gastridiota is a monobasic genus with a relictual distribution in subtropical forests of eastern Australia. The Oriental and African genera comprised a monophyletic group: the Oriental region was inferred to have been colonized from a long-distance dispersal event from Australia to South-East Asia c. 53 Mya or possibly later (c. 36-26 Mya); Africa was subsequently colonized by dispersal from Asia c. 16 Mya (95 % credibility interval: 21-12 Mya). Based on the strongly supported phylogenetic relationships and estimates of divergence times, we conclude that Bombycidae had its origin in the fragment of Southern Gondwana consisting of Australia, Antarctica and South America during the Paleocene. The disjunction between South America (Epiinae) and Australia (Bombycinae) is best explained by vicariance in the Eocene, whereas the disjunct distribution in Asia and Africa is best explained by more recent dispersal events.

Phylogeny and divergence time estimation of Io moths and relatives (Lepidoptera: Saturniidae: Automeris).

The saturniid moth genus Automeris includes 145 described species. Their geographic distribution ranges from the eastern half of North America to as far south as Peru. Automeris moths are cryptically colored, with forewings that resemble dead leaves, and conspicuously colored, elaborate eyespots hidden on their hindwings. Despite their charismatic nature, the evolutionary history and relationships within Automeris and between closely related genera, remain poorly understood. In this study, we present the most comprehensive phylogeny of Automeris to date, including 80 of the 145 described species. We also incorporate two morphologically similar hemileucine genera, Pseudautomeris and Leucanella, as well as a morphologically distinct genus, Molippa. We obtained DNA data from both dry-pinned and ethanol-stored museum specimens and conducted Anchored Hybrid Enrichment (AHE) sequencing to assemble a high-quality dataset for phylogenetic analysis. The resulting phylogeny supports Automeris as a paraphyletic genus, with Leucanella and Pseudautomeris nested within, with the most recent common ancestor dating back to 21 mya. This study lays the foundation for future research on various aspects of Automeris biology, including geographical distribution patterns, potential drivers of speciation, and ecological adaptations such as antipredator defense mechanisms.

Phylogenomic data exploration with increased sampling provides new insights into the higher-level relationships of butterflies and moths (Lepidoptera).

A robust and stable phylogenetic framework is a fundamental goal of evolutionary biology. As the third largest insect order in the world following Coleoptera and Diptera, Lepidoptera (butterflies and moths) play a central role in almost every terrestrial ecosystem as indicators of environmental change and serve as important models for biologists exploring questions related to ecology and evolutionary biology. However, for such a charismatic insect group, the higher-level phylogenetic relationships among its superfamilies are still poorly resolved. Compared to earlier phylogenomic studies, we increased taxon sampling among Lepidoptera (37 superfamilies and 68 families containing 263 taxa) and acquired a series of large amino-acid datasets from 69,680 to 400,330 for phylogenomic reconstructions. Using these datasets, we explored the effect of different taxon sampling with significant increases in the number of included genes on tree topology by considering a series of systematic errors using maximum-likelihood (ML) and Bayesian inference (BI) methods. Moreover, we also tested the effectiveness in topology robustness among the three ML-based models. The results showed that taxon sampling is an important determinant in tree robustness of accurate lepidopteran phylogenetic estimation. Long-branch attraction (LBA) caused by site-wise heterogeneity is a significant source of bias giving rise to unstable positions of ditrysian groups in phylogenomic reconstruction. Phylogenetic inference showed the most comprehensive framework to reveal the relationships among lepidopteran superfamilies, and presented some newly relationships with strong supports (Papilionoidea was sister to Gelechioidea and Immoidea was sister to Galacticoidea, respectively), but limited by taxon sampling, the relationships within the species-rich and relatively rapid radiation Ditrysia and especially Apoditrysia remain poorly resolved, which need to increase taxon sampling for further phylogenomic reconstruction. The present study demonstrates that taxon sampling is an important determinant for an accurate lepidopteran tree of life and provides some essential insights for future lepidopteran phylogenomic studies.

Genome sequencing and assembly of Indian golden silkmoth, Antheraea assamensis Helfer (Saturniidae, Lepidoptera).

Muga silkworm (Antheraea assamensis), one of the economically important wild silkmoths, is unique among saturniid silkmoths. It is confined to the North-eastern part of India. Muga silk has the highest value among the other silks. Unlike other silkmoths, A. assamensis has a low chromosome number (n = 15), and ZZ/ZO sex chromosome system. Here, we report the first high-quality draft genome of A. assamensis, assembled by employing the Illumina and PacBio sequencing platforms. The assembled genome of A. assamensis is 501.18 Mb long, with 2697 scaffolds and an N50 of 683.23 Kb. The genome encompasses 18,385 protein-coding genes, 86.29% of which were functionally annotated. Phylogenetic analysis of A. assamensis revealed its divergence from other Antheraea species approximately 28.7 million years ago. Moreover, an investigation into detoxification-related gene families, CYP450, GST, and ABC-transporter, revealed a significant expansion in A. assamensis as compared to the Bombyx mori. This expansion is comparable to Spodoptera litura, suggesting adaptive responses linked to the polyphagous behavior observed in these insects. This study provides valuable insights into the molecular basis of evolutionary divergence and adaptations in muga silkmoth. The genome assembly reported in this study will significantly help in the functional genomics studies on A. assamensis and other Antheraea species along with comparative genomics analyses of Bombycoidea insects.

Phylogenomics of the geometrid tribe Palyadini (Lepidoptera: Geometridae) reveals contrasting patterns of phylogenetic signal in wing colour characters.

Next generation sequencing techniques currently represent a practical and efficient way to infer robust evolutionary hypotheses. Palyadini is a small Neotropical tribe of geometrid moths composed of six genera that feature strikingly colourful wings. Here, we investigated patterns of evolution and amount of phylogenetic signal contained in various colour characters featured in the wings of members of this tribe by (i) inferring a robust phylogenetic hypothesis using ultraconserved elements (UCEs), and afterwards, (ii) mapping the morphological characters onto the molecular topology under a parsimonious ancestral character optimization. Our matrix, obtained with 60% completeness, includes 754 UCE loci and 73 taxa (64 ingroup, nine outgroup). Maximum likelihood and parsimony generated largely identical topologies with strongly supported nodes, except for one node inside the genus Opisthoxia. According to our topology, most wing colour characters are reconstructed as homoplastic, particularly at the tribe level, but five of the seven provide evidence supporting common ancestry at the genus level. Our results emphasize, once again, that no character system is infallible, and that more research is necessary to take our understanding of the evolution of wing colour in moths to a level comparable with the knowledge we have for butterflies.

Description of Antaeola rhinosa sp. nov. (Lepidoptera: Tortricidae: Olethreutinae) from Thailand.

The genus Antaeola has remained monotypic since its description by Diakonoff in 1973. A new species, Antaeola rhinosa sp. nov., is described and illustrated from Thailand. The new species can be distinguished from A. antaea (Meyrick) by facies and features of the male and female genitalia.

Anurocampa (Lepidoptera: Notodontidae): two new species, systematics and immature stages.

Notodontidae (Lepidoptera, Noctuoidea) comprise over 4,000 described species distributed worldwide, among which nearly half are restricted to the Neotropics. Morphology of adults and immatures of Notodontidae have been broadly investigated and many larval, pupal, and adult characters were found to be synapomorphies of subfamilies and tribes. Despite this, the current classification of Notodontidae remains unsettled as most recent classification systems are contradictory due to reliance on incomplete global sampling and, many taxa, especially in the Neotropics, are still informally classified as incertae sedis. Anurocampa Herrich-Shäffer was recently treated as an incertae sedis genus, and immature and adult characters may provide further evidence for its systematic position among the Notodontidae. With this goal in mind, the present study describes the immature stages of Anurocampa mingens Herrich-Shäffer from Brazil and describes two new species in the genus from Costa Rica based on morphology and mitochondrial DNA: Anurocampa markhastingsi Chacón and St Laurent sp. nov. and Anurocampa abelardochaconi Chacón and St Laurent sp. nov. and discusses the systematic position of Anurocampa.

Letrasilta, a new genus for the Striatella cernyi Volynkin species-group with description of a new species from Xizang, China, and a replacement name for the genus group name Striatella Volynkin & S.-Y. Huang, 2019 (Lepidoptera: Erebidae: Arctiinae: Lithosiini).

The name Striatochrista nom. n. is introduced as replacement for Striatella Volynkin & S.-Y. Huang, 2019. A new genus, Letrasilta S.-Y. Huang & Volynkin gen. n. is erected to include the Striatella cernyi species-group with the new species, L. ratnasambhava S.-Y. Huang, Volynkin & Yin sp. n. from Xizang, southwestern China designated as the type species. Based on the molecular phylogenetic analysis, the new genus is found to be sister to the clade (Aberrasine + ((Indiania + Idopterum) + Striatochrista nom. n.)) but is distinguished from all the relevant genera by the unique genitalia features. Letrasilta cernyi (Volynkin, 2018) comb. n. is also reported from India for the first time. Adults and genitalia of the aforementioned taxa are illustrated. A checklist of the genus Striatochrista is also provided.

Taxonomic review of the Miltochrista hollowai and M. curvifascia species-groups with descriptions of two new species from India (Erebidae: Arctiinae: Lithosiini).

The Miltochrista hollowai (Kirti & Gill, 2009) and M. curvifascia (Hampson, 1891) species-groups are reviewed, and two new species belonging to the M. hollowai species-group i.e., Miltochrista madathumala sp. nov. and M. kumarkaustubhi sp. nov. are described from South India. The female of M. paraarcuata is illustrated and diagnosed for the first time. Illustrations of adults and genitalia as well as diagnoses are provided for all the species considered.

Integrative taxonomic revision of the African taxa of the Racotis Moore, 1887 generic complex (Lepidoptera, Geometridae, Ennominae, Boarmiini).

The Afrotropical taxa of the Racotis s.l. generic complex are revised utilising integrative taxonomical methods. Based on the evaluation of genital morphology and analyses of DNA barcodes, a new genus, Afroracotis gen. n. is established to include the Afrotropical "Racotis" species. The new genus is subdivided into 5 subgenera: Afroracotis subgen. n., Herbuloracotis subgen. n., Rwandaracotis subgen. n., Zebracotis subgen. n. and Sokokeracotis subgen. n.. A new monotypic genus is described to include Boarmia ugandaria Swinhoe, 1904 which was combined earlier with the genus Chorodna and recently with Racotis: Chorocotis gen. n.. Two species formerly assigned to Cleora are moved to Afroracotis: A. albitrigonis (Prout, 1927) comb. n., A. atriclava (Prout, 1926) comb. n.. Two species are transferred from Racotis to Colocleora: C. breijeri (Prout, 1922) comb. n., C. incauta (Prout, 1916) comb. n.. Seventeen new Afroracotis species (A. aliena, A. stadiei, A. violetteae, A. fiebigi, A. turlini, A. dargei, A. longicornuta, A. aristophanousi, A. muscivirens, A. chaineyi, A. lydiae, A. smithi, A. ochsei, A. milesi, A. helicalis, A. takanoi and A. staudei spp. n.) and 5 new subspecies (A. squalida thomensis, A. argillacea morettoi, A. longicornuta congolana, A. longicornuta ugandana and A. lydiae orientalis sspp. n.) are described, totalling 27 species and 8 subspecies contained in the genus Afroracotis. Adults and genitalia of all taxa are illustrated in 210 colour and 129 black and white figures demonstrating the intraspecific variability. The distribution of all taxa is illustrated in 6 dot maps. The results of the genetic analyses are figured in four phylograms.

Description of Sphaleroptera orientana meridionalis subs. n. (Lepidoptera: Tortricidae: Cnephasiini) from the Pale di San Martino Mountain plateau (Dolomites, NE Italy).

On the base of newly collected material, a new subspecies of the alpine endemic moth Sphaleroptera orientana Whitebread, 2006 (Lepidoptera: Tortricidae) from the Pale di San Martino Group (Dolomites, Northern Italy) is described. Morphological characters of the adults and DNA barcode suggests the presence of allopatric populations of S. orientana in the South-eastern Alps, attributable to two distinct subspecies: S. o. suborientana Whitebread, 2006 in the Catinaccio, the Sella, and the Fanes group, and Julian Alps, and S. o. meridionalis subs. nov. known from the Pale of San Martino group. The main biogeographic barrier is constituted by the complex of the Fiemme valleys-Val di Fassa-by the Pordoi Pass and by the Val Cordevole which from west to north to east separate the distribution area of S. o. suorientana from S.o. meridionalis.

New data on the fauna of pyraloid moths (Lepidoptera: Pyraloidea) of Dagestan (Russia).

58 species of pyraloid moths are reported for the East Caucasian region of Russia and the Republic of Dagestan for the first time. Among them, 13 species are new for the fauna of Russia. Additionally, we report first-time records of several pyraloid species from Kazakhstan and also from different regions of Russia: Astrakhan, Volgograd, Voronezh and Orenburg Provinces. Imagoes of four notable species are illustrated.

One new species and two new records of the genus Garudinia Moore, 1882 (Lepidoptera, Erebidae, Arctiinae, Lithosiini) from China.

One new species of the genus Garudinia Moore, 1882, G. ovata sp. nov., is described from China. Two species, G. pseudolatana Holloway, 2001 and G. biplagiata Hampson, 1896, are reported for the first time from China. The female genitalia of G. pseudolatana is described herein for the first time. The new species' imagos and genitalia are illustrated and compared with related species. The distribution map of the genus Garudinia in China is provided.

The known species of the genus Achagua Rindge, 1983, with the description of three new species from the Neotropics (Geometridae: Ennominae).

The previously monotypic geometrid genus Achagua (Ennominae: Nacophorini) is reviewed following the discovery of three undescribed species. New species are as follows: A. cooperae n. sp. described from Costa Rica, A. magna n. sp. described from Ecuador, Peru, and Bolivia; and Achagua velata n. sp. described from French Guiana. Available COI barcode data is briefly reviewed, and adult and genitalic illustrations are provided for each of the four known species.

The first description of the female of Tritonaclia kefersteinii (Butler, 1882)(Lepidoptera: Erebidae: Arctiinae: Syntomini), the type species of its genus.

The previously unknown female of Tritonaclia kefersteinii (Butler, 1882), the type species of the genus Tritonaclia Hamspon, 1898 is described for the first time, based on a specimen from Marojejy National Park (North-east Madagascar). The female genitalia are described in detail and illustrated. The taxonomic affiliation of the newly discovered female and its conspecificity with the male are confirmed by both morphological and molecular characters. For the first time, clear diagnostic characters of the male genitalia are also provided and illustrated. Comments on the known distribution of the species are provided. The new findings allow the genus to be properly characterized as a contribution to the taxonomy of Madagascar Syntomini.

Anti-bat ultrasound production in moths is globally and phylogenetically widespread.

Warning signals are well known in the visual system, but rare in other modalities. Some moths produce ultrasonic sounds to warn bats of noxious taste or to mimic unpalatable models. Here, we report results from a long-term study across the globe, assaying moth response to playback of bat echolocation. We tested 252 genera, spanning most families of large-bodied moths, and document anti-bat ultrasound production in 52 genera, with eight subfamily origins described. Based on acoustic analysis of ultrasonic emissions and palatability experiments with bats, it seems that acoustic warning and mimicry are the raison d'être for sound production in most moths. However, some moths use high-duty-cycle ultrasound capable of jamming bat sonar. In fact, we find preliminary evidence of independent origins of sonar jamming in at least six subfamilies. Palatability data indicate that jamming and warning are not mutually exclusive strategies. To explore the possible organization of anti-bat warning sounds into acoustic mimicry rings, we intensively studied a community of moths in Ecuador and, using machine-learning approaches, found five distinct acoustic clusters. While these data represent an early understanding of acoustic aposematism and mimicry across this megadiverse insect order, it is likely that ultrasonically signaling moths comprise one of the largest mimicry complexes on earth.

A supertree of Northern European macromoths.

Ecological and life-history data on the Northern European macromoth (Lepidoptera: Macroheterocera) fauna is widely available and ideal for use in answering phylogeny-based research questions: for example, in comparative biology. However, phylogenetic information for such studies lags behind. Here, as a synthesis of all currently available phylogenetic information on the group, we produce a supertree of 114 Northern European macromoth genera (in four superfamilies, with Geometroidea considered separately), providing the most complete phylogenetic picture of this fauna available to date. In doing so, we assess those parts of the phylogeny that are well resolved and those that are uncertain. Furthermore, we identify those genera for which phylogenetic information is currently too poor to include in such a supertree, or entirely absent, as targets for future work. As an aid to studies involving these genera, we provide information on their likely positions within the macromoth tree. With phylogenies playing an ever more important role in the field, this supertree should be useful in informing future ecological and evolutionary studies.

Genome-wide analysis of V-ATPase genes in Plutella xylostella (L.) and the potential role of PxVHA-G1 in resistance to Bacillus thuringiensis Cry1Ac toxin.

The rapid development of insecticide resistance has hampered the use of Bacillus thuringiensis (Bt), a widely used bio-pesticide. Plutella xylostella (L.) is a globally distributed lepidopteran pest of cruciferous vegetables and has developed severe field resistance to the Bt toxin. Vacuolar H+-ATPases (VHA) are multi-subunit complexes and participate in multiple physiological processes. However, the characterization and functional studies of VHA genes are lacking in insects. This study performed a genome-wide analysis and identified 35 VHA gene family members divided into 15 subfamilies in P. xylostella. We cloned a V-ATPase subunit G gene, PxVHA-G1, in our previous midgut transcriptome profiles. Quantitative reverse transcriptase-polymerase chain reaction results showed that PxVHA-G1 was upregulated in the Cry1S1000-resistant strain than in the G88-susceptible strain, and its expression profile revealed that the midgut, Malpighian tubules, and larva stages generally showed high expression levels. RNAi-mediated knockdown of the PxVHA-G1 gene increased the susceptibility of P. xylostella (G88 and Cry1S1000) to Cry1Ac toxin. Our study is the first to explore the role of PxVHA-G1 on regulating Cry1Ac toxicity in P. xylostella, thus, providing new insights into the role of VHAs in the development of Cry1Ac resistance and sustainable development of pest management.