COI Barcodes combined with multilocus data for representative Aporia taxa shed light on speciation in the high altitude Irano-Turanian mountain plateaus (Lepidoptera: Pieridae).

Even though the high plateaus of Qinghai-Tibet and Iran share many faunal elements, the historical biogeography of the species present in this area are not very well understood. We present a complete COI barcode library for Aporia Hübner and a first comprehensive phylogeny for the genus including all known species and majority of subspecies using ten available genes (COI-COII, ND1, ND5, Cytb, EF-1a, Wg, 16S, 28S-D2/D3 and 28S-D8). We then focus on A. leucodice (Eversmann, 1843) and related taxa in order to resolve some long-standing taxonomic issues in this species-group. Based on DNA sequence data as well as morphology, we raise Aporia illumina (Grum-Grshimailo 1890) stat. nov. (= pseudoillumina Tshikolovets 2021 syn. nov.) as a distinct species and designate a lectotype; synonymize Aporia leucodice leucodice Eversmann, 1843 (= A. l. morosevitshae Sheljuzhko, 1908 syn. nov.); and describe a new species, Aporia ahura sp. nov., from the Central Alborz Mountains in northern Iran.

DNA barcodes from over-a-century-old type specimens shed light on the taxonomy of a group of rare butterflies (Lepidoptera: Nymphalidae: Calinaginae).

We analyzed COI barcode sequences from 138 over-a-century old specimens of Calinaga including 36 name-bearing type specimens stored at the Natural History Museum London. These new data, combined with previously available RPS5 sequences, divide the Calinaga samples into four well-supported mitochondrial lineages that together with a novel wing-pattern analysis, support the recognition of six species (lhatso, buddha, brahma, aborica, formosana and davidis), with all other names subsumed either as subspecies or synonyms. One new taxon is described, Calinaga aborica naima Vane-Wright, ssp. n.

Phylogenetic and Comparative Genomics Study of Papilionidae Based on Mitochondrial Genomes.

Most species of Papilionidae are large and beautiful ornamental butterflies. They are recognized as model organisms in ecology, evolutionary biology, genetics, and conservation biology but present numerous unresolved phylogenetic problems. Complete mitochondrial genomes (mitogenomes) have been widely used in phylogenetic studies of butterflies, but mitogenome knowledge within the family Papilionidae is limited, and its phylogeny is far from resolved. In this study, we first report the mitogenome of Byasa confusa from the subfamily Papilioninae of Papilionidae. The mitogenome of B. confusa is 15,135 bp in length and contains 13 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes, and an AT-rich control region (CR), closely mirroring the genomic structure observed in related butterfly species. Comparative analysis of 77 Papilionidae mitogenomes shows gene composition and order to be identical to that of an ancestral insect, and the AT bias, Ka/Ks, and relative synonymous codon usage (RSCU) are all consistent with that of other reported butterfly mitogenomes. We conducted phylogenetic analyses using maximum-likelihood (ML) and Bayesian-inference (BI) methods, with 77 Papilionidae species as ingroups and two species of Nymphalidae and Lycaenidae as outgroups. The phylogenetic analysis indicated that B. confusa were clustered within Byasa. The phylogenetic trees show the monophyly of the subfamily Papilioninae and the tribes Leptocircini, Papilionini, and Troidini. The data supported the following relationships in tribe level on Papilioninae: (((Troidini + Papilionini) + Teinopalpini) + Leptocircini). The divergence time analysis suggests that Papilionidae originated in the late Creataceous. Overall, utilizing the largest number of Papilionidae mitogenomes sequenced to date, with the current first exploration in a phylogenetic analysis on Papilionidae (including four subfamilies), this study comprehensively reveals the mitogenome characteristics and mitogenome-based phylogeny, providing information for further studies on the mitogenome, phylogeny, evolution, and taxonomic revision of the Papilionidae family.

Molecular phylogeny of Hesperiidae (Lepidoptera) with an emphasis on Asian and African genera.

Despite considerable research efforts in recent years, the deeper phylogenetic relationships among skipper butterflies (Hesperiidae) remain unresolved. This is primarily because of limited sampling, especially within Asian and African lineages. In this study, we consolidated previous data and extensively sampled Asian and African taxa to elucidate the phylogenetic relationships within Hesperiidae. The molecular dataset comprised sequences from two mitochondrial and two nuclear gene regions from 563 species that represented 353 genera. Our analyses revealed seven subfamilies within Hesperiidae: Coeliadinae, Euschemoninae, Eudaminae, Pyrginae, Heteropterinae, Trapezitinae, and Hesperiinae. The systematics of most tribes and genera aligned with those of prior studies. However, notable differences were observed in several tribes and genera. Overall, the position of taxa assigned to incertae sedis in Hesperiinae is largely clarified in this study. Our results strongly support the monophyly of the tribe Tagiadini (Pyrginae), and the systematics of some genera are clarified with comprehensive discussion. We recognize 15 tribes within the subfamily Hesperiinae. Of these, nine tribes are discussed in detail: Aeromachini, Astictopterini, Erionotini, Unkanini (new status), Ancistroidini, Ismini (confirmed status), Plastingini (new status), Gretnini (confirmed status), and Eetionini (confirmed status). We propose four subtribes within Astictopterini: Hypoleucina subtrib.n., Aclerosina, Cupithina, and Astictopterina. Furthermore, we describe a new genus (Hyarotoidesgen.n.) and reinstate two genera (Zeareinst.stat. and Separeinst.stat.) as valid. Additionally, we propose several new combinations: Zea mythecacomb.n.,Sepa bononiacomb.n. & reinst.stat., and Sepa umbrosacomb.n. This study, with extensive sampling of Asian and African taxa, greatly enhances the understanding of the knowledge of the skipper tree of life.

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.

Hybrid speciation driven by multilocus introgression of ecological traits.

Hybridization allows adaptations to be shared among lineages and may trigger the evolution of new species1,2. However, convincing examples of homoploid hybrid speciation remain rare because it is challenging to demonstrate that hybridization was crucial in generating reproductive isolation3. Here we combine population genomic analysis with quantitative trait locus mapping of species-specific traits to examine a case of hybrid speciation in Heliconius butterflies. We show that Heliconius elevatus is a hybrid species that is sympatric with both parents and has persisted as an independently evolving lineage for at least 180,000 years. This is despite pervasive and ongoing gene flow with one parent, Heliconius pardalinus, which homogenizes 99% of their genomes. The remaining 1% introgressed from the other parent, Heliconius melpomene, and is scattered widely across the H. elevatus genome in islands of divergence from H. pardalinus. These islands contain multiple traits that are under disruptive selection, including colour pattern, wing shape, host plant preference, sex pheromones and mate choice. Collectively, these traits place H. elevatus on its own adaptive peak and permit coexistence with both parents. Our results show that speciation was driven by introgression of ecological traits, and that speciation with gene flow is possible with a multilocus genetic architecture.

Convergent morphology and divergent phenology promote the coexistence of Morpho butterfly species.

The coexistence of closely-related species in sympatry is puzzling because ecological niche proximity imposes strong competition and reproductive interference. A striking example is the widespread wing pattern convergence of several blue-banded Morpho butterfly species with overlapping ranges of distribution. Here we perform a series of field experiments using flying Morpho dummies placed in a natural habitat. We show that similarity in wing colour pattern indeed leads to interspecific territoriality and courtship among sympatric species. In spite of such behavioural interference, demographic inference from genomic data shows that sympatric closely-related Morpho species are genetically isolated. Mark-recapture experiments in the two most closely-related species unravel a strong temporal segregation in patrolling activity of males. Such divergence in phenology reduces the costs of reproductive interference while simultaneously preserving the benefits of convergence in non-reproductive traits in response to common ecological pressures. Henceforth, the evolution of multiple traits may favour species diversification in sympatry by partitioning niche in different dimensions.

Genomics Reveal Admixture and Unexpected Patterns of Diversity in a Parapatric Pair of Butterflies.

We studied the evolutionary relationship of two widely distributed parapatric butterfly species, Melitaea athalia and Melitaea celadussa, using the ddRAD sequencing approach, as well as genital morphology and mtDNA data. M. athalia was retrieved as paraphyletic with respect to M. celadussa. Several cases of mito-nuclear discordance and morpho-genetic mismatch were found in the contact zone. A strongly diverged and marginally sympatric clade of M. athalia from the Balkans was revealed. An in-depth analysis of genomic structure detected high levels of admixture between M. athalia and M. celadussa at the contact zone, though not reaching the Balkan clade. The demographic modelling of populations supported the intermediate genetic make-up of European M. athalia populations with regards to M. celadussa and the Balkan clade. However, the dissimilarity matrix of genotype data (PCoA) suggested the Balkan lineage having a genetic component that is unrelated to the athalia-celadussa group. Although narrowly sympatric, almost no signs of gene flow were found between the main M. athalia group and the Balkan clade. We propose two possible scenarios on the historical evolution of our model taxa and the role of the last glacial maximum in shaping their current distribution. Finally, we discuss the complexities regarding the taxonomic delimitation of parapatric taxa.

A Contribution Towards Resolving the Systematics of the High-Altitude Tropical Andean Satyrine Genus Altopedaliodes Forster, 1964 (Lepidoptera, Nymphalinae: Satyrinae).

The first phylogenetic hypotheses for the high Andean satyrine butterfly genus Altopedaliodes is proposed based on sequence data from mitochondrial (COI and COII) and nuclear (EF-1α) genes. Four species previously included in the genus were found not to be closely related to the clade containing the type species for Altopedaliodes, and these species are therefore removed and placed in the appropriate genus: Pedaliodes cocytia (C. Felder and R. Felder, 1867) n. comb., Pedaliodes nebris Thieme 1905rev. comb., Neopedaliodes tamaensis (Pyrcz and Viloria 2007) n. comb., and Altopedaliodes similis Henao, Páez and Rodríguez-M., 2017; the last name is here synonymized with Pedaliodes nebris (Thieme 1905) n. syn. A taxonomic revision of the twelve species in Altopedaliodes as newly circumscribed is presented and all the currently recognized taxa are revised. We describe two new species, A. llanganati Padrón, Pyrcz and Willmott n. sp. (Ecuador, Napo) and A. pilimbala Pyrcz and Boyer n. sp. (Colombia, Cauca), and three new subspecies, A. reissi papallacta Padrón, Pyrcz and Willmott n. ssp. (Ecuador, Napo), A. reissi dominica Pyrcz and Padrón n. ssp. (Colombia, Valle del Cauca) and A. halli cagnoni Boyer, Pyrcz and Padrón n. ssp. (Ecuador, Morona-Santiago). A new status is proposed for A. nucea Pyrcz and Viloria 1999, n. stat., we combine Altopedaliodes scydmaena (Hayward, 1968) n. comb. and Altopedaliodes belmira (Pyrcz and Rodríguez, 2004), n. comb., n. stat. into the genus and treat the latter taxon as a species, and we newly treat Altopedaliodes scydmaena kruegeri Pyrcz 1999n. stat. as a subspecies.

Toward standards in practices and techniques on ootaxonomy in the Pieridae (Lepidoptera: Papilionoidea).

We present a proposal on the standards used on ootaxonomy practices and techniques in the butterfly family Pieridae Duponchel (Lepidoptera: Papilionoidea) in five stages: 1) getting the specimens, 2) integration into a collection, 3) dissections to recover the exochorion, 4) elaboration of images of it, and 5) the preparation of its description with the necessary diagrams and tabulations. Also, we present the detailed techniques applied in observation and graphic representation, based on the methylene blue staining techniques and those required for the use with the scanning electron microscope (SEM). We compare the result of the standardized techniques with those from macro photography, drawings, and photographs with SEM-all of them found in books with descriptions and images of eggs of the Pieridae. We present a glossary and general aspects of the exochorion in the Pieridae as an Appendix to this article. Standardized techniques show more accurate and extensive character retrieval for systematics. For the scale in which they work, these techniques recovered more information than those present from oviposited eggs, where the exochorionic base is not seen. Also, the descriptions contain detailed data on more structures-which are comparable to each other-than are absent in the references mentioned. We present the recovered characters with the techniques found in the literature as three synthetically supplementary materials.

Alpha-taxonomy and phylogeny of African Junoniini butterflies based on morphological data, with an emphasis on genitalia, and COI barcode (Lepidoptera: Nymphalidae).

The tribe Junoniini is a predominantly Paleotropical group of the cosmopolitan butterfly subfamily Nymphalinae (Nymphalidae), with highest diversity in the Afrotropical region. Its systematics and relationships are not entirely resolved. Question marks remain concerning the validity of some genera; and the apparently close relationship between the Indo-Australian genus Yoma and the Afrotropical Protogoniomorpha, as evidenced by molecular phylogenies, remains a puzzle. Here, we present a cladistic analysis, based on 42 characters of the male and female genitalia of 41 species of Junoniini belonging to six genera, nearly all of them continental Afrotropical, and 3 species of two Indo-Australian genera Yoma and Rhinopalpa. A ML COI-based tree is produced for 36 species of Afrotropical Junoniini and Yoma. The molecular data are consistent with previous studies. However, morphological analysis does not confirm a close relationship between Protogoniomorpha and Yoma. Despite the evolution of a number of modifications, the male genitalia within all genera and species of the Junoniini share a cohesive build plan, in particular a transformed sacculus, from which Yoma is highly divergent. The position of the genus Kamilla, previously synonymized with Junonia, is discussed. Three East African coast taxa, Junonia elgiva stat. reinst., Protogoniomorpha nebulosa stat. reinst. and Salamis amaniensis stat. reinst., and one from central Africa, Precis silvicola stat. reinst. are raised to species level, based on comparative analysis of their male genitalia.

Two new species of Manerebia Staudinger from paramo habitat in the Colombian Eastern Cordillera of the Andes (Lepidoptera: Nymphalidae, Satyrinae, Pronophilina).

Two new species of the genus Manerebia Staudinger, 1897 (Nymphalidae, Satyrinae) are described from paramo habitat on the eastern slopes of the Colombian Eastern Cordillera in the area of Pisba and La Colorada: Manerebia bernito n. sp. and Manerebia clarita n. sp. Both are morphologically most similar to Manerebia levana and Manerebia pervaga, and the possible phylogenetic relationship between them is hypothesized. Their habitat is described and a proposal of their conservation status is made. With the addition of the two new species described in this study, a total of 15 species of Manerebia are reported from Colombia. However, a molecular analysis should be carried out to validate the taxonomic status of several species of Manerebia, in particular having in mind a potentially high cryptic diversity of this genus.

Jamides wananga, a new species from Papua New Guinea and Australia (Lepidoptera: Lycaenidae).

Jamides wananga sp. n. is described and illustrated from Madang Province of Papua New Guinea and Dauan Island in the Torres Strait (Queensland, Australia). The new species is similar in appearance to several other Jamides Hübner species in the bochus-group. DNA sequence data and morphology were used to distinguish the nominotypical subspecies found on the New Guinea mainland from J. w. roxina subsp. n. on Dauan Island. Notes on the habitat and behaviour are also provided.

A brief history of Lepidoptera section contributions to Zootaxa.

As a reviewer, John Brown received the first Lepidoptera manuscript submitted to Zootaxa in 2002. Within a year he was persuaded by a colleague to volunteer as its first Lepidoptera section editor. As submissions increased, he realized that he needed assistance, so in July 2005 he enlisted Robert Robbins (U.S. National Museum of National History), and the two editors split the submissions-Brown covered moths and Robbins butterflies. As submissions continued to grow, Robbins stepped down and Brown was again the sole editor. Owing to the ever-increasing manuscript load, in 2007 Brown submitted a proposal to several colleagues, inviting them to become Lepidoptera section editors, with the concept that the more editors there were, the fewer manuscripts each would have to handle, and their duties would include papers primarily in their area of expertise. The solicitation was successful, with four new subject editors coming on board in 2007: Lawrence Gall for macrolepidoptera families, Michael Toliver for butterflies, Jean-François Landry for microlepidoptera families, and Shen-Horn Yen for Pyraloidea and Zygaenoidea; the last two are still section editors today. Over the next 13 years, numerous editors came and went-turnover in editorship was always viewed as a positive way to involve new scientists and interject fresh ideas. From 2001 to 2020, a cumulative total of 21 scientists have served as Lepidoptera Section editors (Table 1), representing 14 different countries.

One Hundred Years of Solitude: rediscovery of Catasticta lycurgus (Godman amp; Salvin, 1880), a yellow butterfly from the Sierra Nevada de Santa Marta, Colombia (Lepidoptera: Pieridae).

Catasticta lycurgus is a striking endemic butterfly, restricted to high elevation habitats in the Sierra Nevada de Santa Marta, an isolated mountain range separate from the Andes in the north of Colombia. The type, which for almost a hundred years was the only known specimen, was collected in 1878 by Frederick Simons in the vicinity of Atánquez and was sent to the UK to be described by renowned naturalists Godman and Salvin in 1880. In 1972, explorers Adams and Bernard collected a second specimen of C. lycurgus in the locality of San Pedro at 2,900m of elevation in the Sierra Nevada de Santa Marta. These two specimens were the only known ones for many decades until recently, when Colombian entomologists found the species again in San Pedro de la Sierra and later, when a female was discovered in 2013. Here, we report the rediscovery of this rare and charismatic species, with new specimens collected near the type locality, which have not been reported previously. The female of C. lycurgus is described and illustrated for the first time as well the male genitalia of this species. We combine all information available to provide some insights on the systematic relationships of this species within the genus Catasticta Butler, discuss its distribution and provide a preliminary conservation assessment. Despite the newly collected specimens, the species remains very rare in the field and in collections.

Taxonomy of the Neolycaena (Rhymnaria) davidi (Oberthür, 1881) species group, with the description of two new taxa from Russia and China (Lepidoptera, Lycaenidae).

Poorly known taxa of the Neolycaena (Rhymnaria) davidi (Oberthür, 1881) species group are reviewed in the light of modern findings. The taxon kozlovi Zhdanko, 1996 previously considered a subspecies of N. (R.) davidi is recognised here as a separate species, N. (R.) kozlovi Zhdanko, 1996, stat. rev., including the subspecies N. (R.) kozlovi namkhaidorji Churkin, 2004, comb. nov., based on morphological differences, the geographic isolation and ecological specialisation. A new species, N. (R.) markhasiovi sp. n., is described from Xinjiang Uyghur Autonomous Region in China (Eastern Tian Shan, the Karlik Mountains) on the basis of distinct genitalia morphology and features of the external morphology, including colouration of the female abdomen. Populations of the subgenus Rhymnaria from southern Tuva in Russia (the Tannu-Ola Mountains) are described as a separate subspecies, N. (R.) davidi tannuola ssp. n. A checklist of the taxa of the N. (R.) davidi species group including data on their distribution and host plants is presented, as well as a key to the species of the group.

Micropylar and Perimicropylar Regions of the Egg Exochorion in Six Genera of Dismorphiinae (Lepidoptera: Pieridae).

Using scanning electron microscopy (SEM), we examine and compare the micropylar and perimicropylar regions of the egg exochorion in six genera of the subfamily Dismorphiinae (Pieridae): Leptidea Billberg, Enantia Hübner, Pseudopieris Godman Salvin, Lieinix Gray, Dismorphia Hübner, and Moschoneura Butler. We provide the first descriptions of the morphological structures of the chorion in these genera. Leptidea is the most primitive genus, and Dismorphia represents the most diverse. We examined the jethys complex of Enantia to study diversity of characters within the genus. We conclude that the exochorion of Dismorphiinae is the most plesiomorphic in relation to Coliadinae and Pierinae, owing to its simple morphology without a transition zone surrounding the wreath (perimicropylar region). Leptidea (Leptideini) shows the least derived characters of the subfamily, followed by Enantia (Dismorphiini). The latter genus exhibits several typical characters of the tribe that are distinctive from other more specialized genera. Exochorionic similarities among the four species of the jethys complex support their close relationship; E. mazai exhibits the most derived features among the species of the complex. Pseudopieris and Moschoneura exhibit several plesiomorphies, such as a large number of openings (micro-grid), but they also show some derived features, such as three semi-rings in the wreath. Dismorphia and Lieinix exhibit characters in their most derived states, such as a striated micro-grid (most conspicuous in Lieinix) and basal aeropyles (most conspicuous in Dismorphia), a character shared with Moschoneura. The purpose of this work is exploratory and descriptive and would be improved by a larger sample size. This contribution is the third in a series of studies on the chorion of Dismorphiinae eggs.

Description of five new species of Epitola sensu lato (Lepidoptera: Lycaenidae: Poritiinae) from West and Central Africa.

Five new species of Epitola s.l. (Lycaenidae: Poritiinae) are described in the genera Stempfferia Jackson, 1962 and Cephetola Libert, 1999. Stempfferia salonga sp. n. was found in the 'Cuvette Centrale' of the Democratic Republic of Congo and a single specimen was identified also from southern Cameroon. The similar S. buea sp. n. is known only from the unique holotype collected at the foothills of Mount Cameroon, while S. michelliberti sp. n. is distributed in lowland rainforest between southeast Ivory Coast and Guinea. The new species of Cephetola were found only in Liberia. Cephetola praecox sp. n. is known from a single upland forest locality in the Putu Range, whereas C. wologizi sp. n. was collected also in upland forest in the Wologizi Mountains.

Geographical distribution of Emesis/ Fabricius (Lepidoptera: Riodinidae) in Mexico: Updated checklist and temporal patterns.

We present a synthesis of the existing information on the genus Emesis Fabricius in Mexico concerning biogeographical patterns and taxonomical aspects. Emesis is the most diverse genus of Emesidini with 57 species and subspecies, with Mexico as the northern limit of this Neotropical genus. We analyzed 5434 specimens of the Lepidoptera Collection of the MZFC, UNAM and compared them with specimens from collections of Mexico, Central and South America. Taxonomic determination and corroboration were made by analysis of wing patterns and genitalia. Geographic distribution and phenology were obtained from the database MARIPOSA. We present an updated list of Emesis of Mexico, with 17 species and subspecies. For each species, we provide information on phenology, geographic, altitudinal, and vegetation distributions. We discuss taxonomic and undersampling concerns for some species, as well as spatial and temporal patterns with special reference to vegetation types and biogeographic provinces in Mexico.