A DNA barcode library of Austrian geometridae (Lepidoptera) reveals high potential for DNA-based species identification.

Situated in the Eastern section of the European Alps, Austria encompasses a great diversity of different habitat types, ranging from alpine to lowland Pannonian ecosystems, and a correspondingly high level of species diversity, some of which has been addressed in various DNA barcoding projects. Here, we report a DNA barcode library of all the 476 species of Geometridae (Lepidoptera) that have been recorded in Austria. As far as possible, species were sampled from different Austrian regions in order to capture intraspecific genetic variation. In total, 2500 DNA barcode sequences, representing 438 species, were generated in this study. For complete coverage of Austrian geometrid species in the subsequent analyses, the dataset was supplemented with DNA barcodes from specimens of non-Austrian origin. Species delimitations by ASAP, BIN and bPTP methods yielded 465, 510 and 948 molecular operational taxonomic units, respectively. Congruency of BIN and ASAP partitions with morphospecies assignments was reasonably high (85% of morphospecies in unique partitions), whereas bPTP appeared to overestimate the number of taxonomic units. The study furthermore identified taxonomically relevant cases of morphospecies splitting and sharing in the molecular partitions. We conclude that DNA barcoding and sequence analysis revealed a high potential for accurate DNA-based identification of the Austrian Geometridae species. Additionally, the study provides an updated checklist of the geometrid moths of Austria.

Patterns of speciation in a parapatric pair of Saturnia moths as revealed by target capture.

The focus of this study has been to understand the evolutionary relationships and taxonomy of a widely distributed parapatric species pair of wild silk moths in Europe: Saturnia pavonia and Saturnia pavoniella (Lepidoptera: Saturniidae). To address species delimitation in these parapatric taxa, target enrichment and mtDNA sequencing was employed alongside phylogenetic, admixture, introgression, and species delimitation analyses. The dataset included individuals from both species close to and farther away from the contact zone as well as two hybrids generated in the lab. Nuclear markers strongly supported both S. pavonia and S. pavoniella as two distinct species, with hybrids forming a sister group to S. pavoniella. However, the Maximum Likelihood (ML) tree generated from mtDNA sequencing data presented a different picture, showing both taxa to be phylogenetically intermixed. This inconsistency is likely attributable to mitonuclear discordance, which can arise from biological factors (e.g., introgressive hybridization and/or incomplete lineage sorting). Our analyses indicate that past introgressions have taken place, but that there is no evidence to suggest an ongoing admixture between the two species, demonstrating that the taxa have reached full postzygotic reproductive isolation and hence represent two distinct biological species. Finally, we discuss our results from an evolutionary point of view taking into consideration the past climatic oscillations that have likely shaped the present dynamics between the two species. Overall, our study demonstrates the effectiveness of the target enrichment approach in resolving shallow phylogenetic relationships under complex evolutionary circumstances and that this approach is useful in establishing robust and well-informed taxonomic delimitations involving parapatric taxa.

Surprising discovery of an enigmatic geometrid in Croatia: Mirlatiaarcuata, gen. nov., sp. nov. (Lepidoptera, Geometridae).

A new monotypic genus of Geometridae, Mirlatiagen. nov., and a new species, M.arcuatasp. nov., are described from Croatia. Based on external and genitalia characters, the new genus is tentatively placed in the subfamily Larentiinae. However, the new genus takes a highly isolated position by having unique characters of the tympanum and showing an unusually long pectination of female antennae. Genetic analysis of a fragmented DNA barcode (mtDNA; cytochrome c oxidase 1) did not result in a clear assignation to any geometrid subfamily or tribe. Adults, male and female genitalia, and habitat photos of the type locality of the new species are illustrated.

Thiotrichasumpichi sp. nov. - a new species of Thiotrichinae (Lepidoptera, Gelechiidae) from south-eastern Europe.

Thiotrichasumpichisp. nov. is described from Greece and Croatia. The systematic position of the new species within Thiotricha is discussed based on external and genitalia characters and from DNA barcodes of the mitochondrial COI gene (cytochrome c oxidase 1). Adults, details of external morphology, and male and female genitalia of the new species are illustrated.

A new species of Neurothaumasia Le Marchand (Lepidoptera, Tineidae) from Crete, Greece.

A new species of fungus moths (Tineidae), Neurothaumasia cretica sp. nov., is described from specimens collected on Crete isl. (Greece). It differs from congeneric taxa by the characteristic black and white forewing pattern which is only shared with N. fasciata Petersen, 1959 from the Middle East, and the widespread western Palaearctic N. ankerella (Mann, 1867). However, the new species differs strongly from the former by several characters of male and female genitalia, and from the latter species particularly from external appearance and by the highly divergent DNA barcode (cytochrome c-oxidase subunit 1) (unknown for N. fasciata). Adult and genitalia of N. cretica sp. nov. and the only similar European species N. ankerella are shown for comparison. Finally, a complete checklist of the genus is added.

A surprising new species of Exapate Hübner, 1825 (Lepidoptera, Tortricidae) from Armenia.

A new species of cold-season moths, Exapate aidasi sp. nov., is described from male specimens collected in Armenia. This is also the first record of the genus Exapate in Armenia and south of the Caucasus. The new species differs from the two previously known congeneric taxa both in phenotypic characteristics and in the structure of the male genitalia. In addition, DNA barcodes (cytochrome c-oxidase subunit 1) are clearly divergent from those of other species. Adult and male genitalia of all three species of Exapate are shown for comparison.

Klimeschiopsis terroris auctt. from Spaina further case of cryptic diversity in European Lepidoptera (Lepidoptera, Gelechiidae, Gelechiinae).

A new species of Gnorimoschemini (Gelechiidae), Klimeschiopsis arnoldfransorum sp. nov., is described from specimens collected in Spain. The species is most closely related to Klimeschiopsis terroris (Hartig, 1938), with which it was hitherto mixed, but differs particularly by its small size with reduced yellowish-white forewing markings, the characters of the male and female genitalia, and the highly divergent DNA barcode (cytochrome c-oxidase subunit 1). Adult and genitalia of both sexes are figured. Finally, a checklist of the genus is provided.

Eucosma subvittana (Staudinger 1892) stat. rev., a Mediterranean species resurrected by DNA barcodes and morphology (Lepidoptera, Tortricidae).

Eucosma subvittana is resurrected from synonymy with E. cana (Haworth, 1811) and redescribed from extensive material collected in Greece (Crete) and Tunisia. It is distinguished from the similar E. cana by constant differences in wing pattern and the female genitalia, whereas the male genitalia are inseparable. A lectotype of E. subvittana is designated in order to fix the identity of this species. Adults and genitalia of both species are figured extensively. Furthermore, the DNA barcodes (cytochrome c-oxidase subunit 1) of both species are clearly divergent. In comparison, several cryptic taxa of related species-groups are genetically inseparable.

Sattleria enrosadira sp. nov., a new cryptic, high alpine species from Northern Italy revealed by DNA barcodes and morphology (Lepidoptera, Gelechiidae).

Sattleria karsholti Huemer Hebert, 2011 is re-examined based on recently collected material, museum vouchers, and DNA barcodes. Adult and male genitalia morphology and molecular data of the COI barcode region support the existence of two allopatric species, S. karsholti, from the type locality of Pizo Arera in the Orobian Alps (Bergamo Province, Italy), and S. enrosadira sp. nov. from Monte Baldo, Adamello and the Brenta group (Trento and Verona Provinces, Italy). Adults and male genitalia of both species are figured; the females of both species are unknown.

Integrative taxonomy reveals overlooked cryptic diversity in the conifer feeding Batrachedrapinicolella (Zeller, 1839) (Lepidoptera, Batrachedridae).

During efforts to generate DNA barcodes for all European Lepidoptera, Batrachedrapinicolella (Zeller, 1839) was found to comprise two genetically distinct clusters. Morphological investigation and results from two nuclear markers and ddRAD sequencing furthermore support the existence of two distinct taxa which we treat as two separate species, B.pinicolella and B.confusella sp. nov. A lectotype for B.pinicolella is designated. Available data indicate that the biology of both species also differs, with Piceaabies (L.) Karsten as a proved host-plant for B.pinicolella and Pinussylvestris L. for B.confusella sp. nov. Both species are mainly distributed on the European continent with B.pinicolella occurring in boreal parts of North and Central Europe and introduced to Canada, reflecting a boreo-montane distribution pattern. Batrachedraconfusella sp. nov. is more widely distributed in temperate Northern and Central Europe.

New species and new records of the genus Filatima Busck, 1939 (Lepidoptera, Gelechiidae) from Central Asia.

Four new species of Filatima Busck, 1939 are described from Central Asia: Filatimaarmata sp. nov. (Iran), F.subarmata sp. nov. (Pakistan, Iran), F.afghana sp. nov. (Afghanistan), and F.karii sp. nov. (Tajikistan). The hitherto unknown female of Filatimamulticornuta Bidzilya & Nupponen, 2018 is described. Recorded to occur for the first time are Filatimatextorella (Chrétien, 1908) from North Macedonia and Turkey, F.pallipalpella (Snellen, 1884) from Kyrgyzstan, and Filatimazagulajevi Anikin & Piskunov, 1996 from Kazakhstan. Filatimafontisella Lvovsky & Piskunov, 1989 is removed from the list of Russian Gelechiidae due to re-identification of the only record as F.multicornuta. An annotated checklist of Palaearctic Filatima species is provided.

Underestimated cryptic diversity in the Caryocolumtricolorella species complex (Lepidoptera, Gelechiidae).

The taxonomy of the Caryocolumtricolorella species complex, an informal subsection of the diverse Caryocoluminteralbicella species group, is revised and four species are separated from DNA barcodes of the mitochondrial COI (cytochrome c oxidase subunit 1) gene and adult morphology: C.tricolorella (Haworth, 1812), C.fibigerium Huemer, 1988, C.herwigvanstaai sp. nov., and C.olekarsholti sp. nov. These species show a vicariant distribution pattern, with C.tricolorella widely distributed in Central and Northern Europe, C.fibigerium restricted to the Iberian Peninsula and southern France, C.herwigvanstaai sp. nov. to the Italian Peninsula, and C.olekarsholti sp. nov. to the Balkans. All species are described in detail, and the adults and genitalia of both sexes are illustrated.

Taxonomy and faunistics of the genus Scrobipalpa Janse, 1951 (Lepidoptera, Gelechiidae) in southern Siberia.

Thirteen new species of Scrobipalpa Janse, 1951 are described: S. nupponeni sp. nov. (Russia: Buryatia, Altai), S. rutjani sp. nov. (Kyrgyzstan; Russia: Novosibirsk reg., Tuva), S. tannuolella sp. nov. (Russia: Tuva), S. kullbergi sp. nov. (Russia: Tuva), S. lobata sp. nov. (Russia: southern Ural, Altai, Tuva, Buryatia), S. inexplictata sp. nov. (Russia: Altai; Armenia), S. altubura sp. nov. (Russia: Altai, Tuva, Buryatia), S. krasnogorka sp. nov. (Russia: Altai), S. buryatica sp. nov. (Russia: Buryatia), S. albiflava sp. nov. (Russia: Altai, Tuva), S. ravida sp. nov. (Russia: Buryatia, Zabaikalskiy krai), S. ochronerva sp. nov. (Russia: Buryatia) and S. ochraeata sp. nov. (Russia: Zabaikalskiy krai). Four new generic combinations are proposed: Scrobipalpa saxatilis (Povolný, 1996) comb. nov.; Scrobipalpa truncata (Povolný, 2002) comb. nov.; Scrobipalpa superba (Povolný, 1996) comb. nov. and Scrobipalpa cinerea (Povolný, 1996) comb. nov. Five new synonyms are established: S. punctata (Povolný, 1996) syn. nov. of S. zaitzevi Piskunov, 1990; S. abstrusa Huemer & Karsholt, 2010 syn. nov. of S. truncata (Povolný, 2002); S. pinosa (Povolný, 2001) syn. nov. of S. ferruginosa (Povolný, 2001); S. kyrana (Povolný, 2001) syn. nov. of S. similis Povolný, 1973 and S. zouhari Povolný, 1984 syn. nov. of S. rebeli (Preissecker, 1915). Scrobipalpa sibirica Bidzilya, 2009 sp. rev., stat. nov. is taken out from synonymy with S. occulta (Povolný, 2002) and S. japonica Povolný, 1977 sp. rev., stat. nov. is taken out from synonymy with S. rebeli (Preissecker, 1914). Six species are recorded from Russia for the first time: Scrobipalpa superstes Povolný, 1977; S. punctulata Li & Bidzilya, 2019; S. strictella Bidzilya & Li, 2010; S. inferna Povolný, 1973; S. mongolica Povolný, 1969 and S. mixta Huemer & Karsholt, 2010. Scrobipalpa notata (Povolný, 2001) is new for Armenia, S. frugifera Povolný, 1969 and S. mixta are new for Kazakhstan, S. acuta (Povolný, 2001) and S. ferruginosa are new for Mongolia, S. tenebrata (Povolný, 2001) is new for China, S. zaitzevi is new for Tajikistan and Afghanistan, S. truncata is new for Kyrgyzstan and Mongolia, S. marmorella Povolný, 1969 and S. candicans (Povolný, 1996) are new for Tajikistan. The male genitalia of S. ferruginosa, and the female genitalia of S. bidzilyai (Povolný, 2001), S. chitensis (Povolný, 2001), S. tenebrata, S. cinerosella Bidzilya, 2009 and S. japonica are described for the first time. Improved diagnosis and additions to descriptions are provided for some little known and poorly diagnosed species. Adults and genitalia are illustrated for all new species and for most of the species which represent new faunistic records.

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.

Gelechiaomelkoi sp. nov. - a new species from the Russian Altai Mountains related to the Nearctic Gelechiamandella Busck, 1904 (Lepidoptera, Gelechiidae), with a synopsis of Gelechia from the Altai Republic of Russia.

Gelechiaomelkoi sp. nov. is described from the Ukok plateau and South Chuisky ridge in the Altai Mountains of Russia. The adult of the new species, including its male genitalia, is illustrated and compared with species most similar in morphology and DNA barcodes-G.sororculella (Hübner, 1817) and G.jakovlevi Krulikovsky, 1905 from the Palaearctic region, as well as G.mandella Busck, 1904 from Canada. This last species is redescribed based on adult specimens, including the genitalia of both sexes, and a lectotype is designated. Gelechiasirotina Omelko, 1986 is recorded from the Altai Republic for the first time. An updated list of six species of Gelechia from the Altai Mountains of Russia is given. Dorsal habitus photographs of all species are provided. The male genitalia of the lectotype of G.jakovlevi is illustrated for the first time.

High resolution DNA barcode library for European butterflies reveals continental patterns of mitochondrial genetic diversity.

The study of global biodiversity will greatly benefit from access to comprehensive DNA barcode libraries at continental scale, but such datasets are still very rare. Here, we assemble the first high-resolution reference library for European butterflies that provides 97% taxon coverage (459 species) and 22,306 COI sequences. We estimate that we captured 62% of the total haplotype diversity and show that most species possess a few very common haplotypes and many rare ones. Specimens in the dataset have an average 95.3% probability of being correctly identified. Mitochondrial diversity displayed elevated haplotype richness in southern European refugia, establishing the generality of this key biogeographic pattern for an entire taxonomic group. Fifteen percent of the species are involved in barcode sharing, but two thirds of these cases may reflect the need for further taxonomic research. This dataset provides a unique resource for conservation and for studying evolutionary processes, cryptic species, phylogeography, and ecology.

Megacraspedus laseni sp. nov. (Lepidoptera: Gelechiidae) from the Dolomites of north-eastern Italy.

Megacraspedus laseni sp. nov. is described from Dolomiti Bellunesi (Veneto Region, Prov. Belluno, Italy). The habitus of the adult and male genitalia are described and illustrated whereas the female sex remains unknown. The new species belongs to the Megacraspedus pentheres species group and is closely related to the southern alpine M. eburnellus Huemer Karsholt, 2001 from which it differs in morphological characters and in DNA barcode sequence. Megacraspedus carolustertius Gastón Vives, 2020 is synonymized with M. quadristictus Lhomme, 1946, syn. nov.

Identity of some recently described Lepidoptera from France-re-assessed with DNA barcodes and morphology.

Seventy-three species of Lepidoptera described from France since 2000, particularly by Jacques Nel and Thierry Varenne, are re-assessed from largely unpublished molecular data. We tried to obtain DNA barcode sequences from 62 holotypes, supplemented by paratypes of eight species and on one case by non-type material, whereas one previously synonymized species was not sequenced. Altogether we obtained 78 DNA barcode sequences for 65 nominal taxa while sequencing failed for six holotypes. An integrative analysis from molecular data and morphology supports the validity of the majority of species but also resulted in the re-assessment of several taxa. The following 13 new synonymies are established: Stigmella cyrneorolandi Nel Varenne, 2013 syn. nov. of Stigmella rolandi van Nieukerken, 1990; Stigmella thibaulti Varenne Nel, 2019 syn. nov. of Stigmella nivenburgensis (Preissecker, 1942) (Nepticulidae); Nemapogon peslieri Varenne Nel, 2017 syn. nov. of Nemapogon inexpectata Varenne Nel, 2017 (Tineidae); Phyllonorycter acericorsica Varenne Nel, 2015 syn. nov. of Phyllonorycter ochreojunctella (Klimesch, 1942) (Gracillariidae); Ancylis paraobtusana Varenne, Nel, Peslier, 2020 syn. nov. of Ancylis comptana (Frölich, 1828) (Tortricidae); Celypha paludicolella Varenne Nel, 2017 syn. nov. of Celypha doubledayana (Barrett, 1872) (Tortricidae); Cydia oxytropidana Nel Varenne, 2016 syn. nov. of Cydia oxytropidis (Martini, 1912) (Tortricidae); Sorhagenia orocorsa Varenne Nel, 2016 syn. nov. of Sorhagenia janiszewskae Riedl, 1962 (Cosmopterigidae); Chionodes cerdanica Peslier, Nel Varenne, 2020 syn. nov. of Chionodes distinctella (Zeller, 1839) (Gelechiidae); Elachista bidentata Varenne Nel, 2019 syn. nov. of Elachista orstadii Palm, 1943; Elachista karsticola Varenne Nel, 2018 syn. nov. of Elachista maculosella Chrétien, 1896 (Elachistidae); Scythris chablaisensis Delmas, 2018 syn. nov. of Scythris laminella ([Denis Schiffermüller], 1775) (Scythrididae); Epermenia pumila (Buvat Nel, 2000) syn. nov. of Epermenia profugella (Stainton, 1856) (Epermeniidae). Finally, the status of some taxa still remains unclear due to the lack of DNA barcodes of closely related species and the absence of convincing diagnostic characters in morphology.