A genetic atlas for the butterflies of continental Canada and United States.

Multi-locus genetic data for phylogeographic studies is generally limited in geographic and taxonomic scope as most studies only examine a few related species. The strong adoption of DNA barcoding has generated large datasets of mtDNA COI sequences. This work examines the butterfly fauna of Canada and United States based on 13,236 COI barcode records derived from 619 species. It compiles i) geographic maps depicting the spatial distribution of haplotypes, ii) haplotype networks (minimum spanning trees), and iii) standard indices of genetic diversity such as nucleotide diversity (π), haplotype richness (H), and a measure of spatial genetic structure (GST). High intraspecific genetic diversity and marked spatial structure were observed in the northwestern and southern North America, as well as in proximity to mountain chains. While species generally displayed concordance between genetic diversity and spatial structure, some revealed incongruence between these two metrics. Interestingly, most species falling in this category shared their barcode sequences with one at least other species. Aside from revealing large-scale phylogeographic patterns and shedding light on the processes underlying these patterns, this work also exposed cases of potential synonymy and hybridization.

Additional taxonomic refinements suggested by genomic analysis of butterflies.

Comparative analyses of genomic data reveal further insights into the phylogeny and taxonomic classification of butterflies presented here. As a result, 2 new subgenera and 2 new species of Hesperiidae are described: Borna Grishin, subgen. n. (type species Godmania borincona Watson, 1937) and Lilla Grishin, subgen. n. (type species Choranthus lilliae Bell, 1931) of Choranthus Scudder, 1872, Cecropterus (Murgaria) markwalkeri Grishin, sp. n. (type locality in Mexico: Sonora), and Hedone yunga Grishin, sp. n. (type locality in Bolivia: Yungas, La Paz). The lectotype is designated for Aethilla toxeus Plötz, 1882. The type locality of Dion uza (Hewitson, 1877) is likely in southern Brazil. A number of taxonomic changes are proposed. The following taxa are subgenera, not genera: Plebulina Nabokov, 1945 of Icaricia Nabokov, 1945; Sinia Forster, 1940 of Glaucopsyche Scudder, 1872; Pseudophilotes Beuret, 1958 of Palaeophilotes Forster, 1938; and Agraulis Boisduval & Le Conte, [1835] of Dione Hübner, [1819]. Asbolis Mabille, 1904 is a subgenus of Choranthus Scudder, 1872 rather than its synonym. The following are species, not subspecies or synonyms: Glaucopsyche algirica (Heyne, 1895) (not Glaucopsyche melanops (Boisduval, 1829)), Chlosyne flavula (W. Barnes & McDunnough, 1918) (not Chlosyne palla (Boisduval, 1852)), Cercyonis hypoleuca Hawks & J. Emmel, 1998 (not Cercyonis sthenele (Boisduval, 1852)), Cecropterus coyote (Skinner, 1892) and Cecropterus nigrociliata (Mabille & Boullet, 1912) (not Aethilla toxeus Plötz, 1882), Aguna malia Evans, 1952 (not Aguna megaeles (Mabille, 1888)), Polygonus arizonensis (Skinner, 1911), Polygonus histrio Röber, 1925, Polygonus pallida Röber, 1925, and Polygonus hagar Evans, 1952 (not Polygonus leo (Gmelin, [1790])), Viola kuma (Bell, 1942), comb. nov. (not Pachyneuria helena (Hayward, 1939)), Tamela maura (Snellen, 1886) (not Tamela othonias (Hewitson, 1878)), Tamela diocles (Moore, [1866]) (not Tamela nigrita (Latreille, [1824])), Vinius phellus (Mabille, 1883) (not Vinius exilis (Plötz, 1883)), Vinius sophistes (Dyar, 1918) (not Vinius tryhana (Kaye, 1914)), and Rhinthon andricus (Mabille, 1895) and Rhinthon aqua (Evans, 1955) (not Rhinthon braesia (Hewitson, 1867)). The following are new and revised species-subspecies combinations: Cercyonis sthenele damei W. Barnes & Benjamin, 1926 (not Cercyonis meadii (W. H. Edwards, 1872)) and Chlosyne flavula blackmorei Pelham, 2008 and Chlosyne flavula calydon (W. Holland, 1931) (not Chlosyne palla). The following are valid subspecies resurrected from synonymy in new and reinstated species-subspecies combinations: Chlosyne palla pola (Boisduval, 1869) (not Chlosyne gabbii gabbii (Behr, 1863)) and Cercyonis meadii mexicana R. Chermock, 1949 (not Cercyonis sthenele damei W. Barnes & Benjamin, 1926, comb. rev.). The following are new junior subjective synonyms: Aethilla toxeus Plötz, 1882 of Cecropterus albociliatus (Mabille, 1877) and Viola dagamba Steinhauser, 1989 of Viola kuma (Bell, 1942), comb. nov., stat. rest. Leucochitonea janice Ehrmann, 1907 is a junior subjective synonym of Heliopetes alana (Reakirt, 1868) and not of Heliopetes petrus (Hübner, [1819]). The holotype of Hermeuptychia sinuosa Grishin, 2021 is illustrated after being spread.

Taxonomic discoveries enabled by genomic analysis of butterflies.

The comparative genomics of butterflies yields additional insights into their phylogeny and classification that are compiled here. As a result, 3 genera, 5 subgenera, 5 species, and 3 subspecies are proposed as new, i.e., in Hesperiidae: Antina Grishin, gen. n. (type species Antigonus minor O. Mielke, 1980), Pompe Grishin and Lamas, gen. n. (type species Lerema postpuncta Draudt, 1923), and Curva Grishin, gen. n. (type species Moeris hyagnis Godman, 1900); in Lycaenidae: Fussia Grishin, subgen. n. (type species Polyommatus standfussi Grum-Grshimailo, 1891) and Pava Grishin, subgen. n. (type species Thecla panava Westwood, 1852); in Hesperiidae: Monoca Grishin, subgen. n. (type species Tagiades monophthalma Plötz, 1884), Putuma Grishin, subgen. n. (type species Tisias putumayo Constantino and Salazar, 2013), and Rayia Grishin, subgen. n. (type species Mastor perigenes Godman, 1900); Cissia wahala Grishin, sp. n. (Nymphalidae; type locality in Mexico: Oaxaca); in Hesperiidae: Hedone mira Grishin and Lamas, sp. n. (type locality in Peru: Apurímac), Vidius pompeoides Grishin, sp. n. (type locality in Brazil: Amazonas), Parphorus hermieri Grishin, sp. n. (Hesperiidae; type locality in Brazil: Rondônia), and Zenis par Grishin, sp. n. (Hesperiidae; type locality in Peru: Cuzco); in Pieridae: Glutophrissa drusilla noroesta Grishin, ssp. n. (type locality in USA: Texas, Cameron Co.) and Pieris marginalis siblanca Grishin, ssp. n. (type locality in USA: New Mexico, Lincoln Co.), and Argynnis cybele neomexicana Grishin, ssp. n. (Nymphalidae; type locality in USA: New Mexico, Sandoval Co.). Acidalia leto valesinoides-alba Reuss, [1926] and Acidalia nokomis valesinoides-alba Reuss, [1926] are unavailable names. Neotypes are designated for Mylothris margarita Hübner, [1825] (type locality in Brazil) and Papilio coras Cramer, 1775 (type locality becomes USA: Pennsylvania, Montgomery Co., Flourtown). Mylothris margarita Hübner, [1825] becomes a junior objective synonym of Pieris ilaire Godart, 1819, currently a junior subjective synonym of Glutophrissa drusilla (Cramer, 1777). Lectotypes are designated for Hesperia ceramica Plötz, 1886 (type locality in Indonesia: Seram Island), Pamphila trebius Mabille, 1891 (type locality Colombia: Bogota), Methionopsis modestus Godman, 1901 and Papias microsema Godman, 1900 (type locality in Mexico: Tabasco), Hesperia fusca Grote & Robinson, 1867 (type locality in USA: Georgia), Goniloba corusca Herrich-Schäffer, 1869, and Goniloba devanes Herrich-Schäffer, 1869; the type localities of the last two species, together with Pamphila stigma Skinner, 1896 and Carystus (Argon) lota (Hewitson, 1877), are deduced to be in South America. Type locality of Junonia pacoma Grishin, 2020 is in Sinaloa, not Sonora (Mexico). Abdomen is excluded from the holotype of Staphylus ascalon (Staudinger, 1876). Furthermore, a number of taxonomic changes are proposed. Alciphronia Koçak, 1992 is treated as a subgenus, not a synonym of Heodes Dalman, 1816. The following genera are treated as subgenera: Lafron Grishin, 2020 of Lycaena [Fabricius], 1807, Aremfoxia Real, 1971 of Epityches D'Almeida, 1938, Placidina D'Almeida, 1928 of Pagyris Boisduval, 1870, and Methionopsis Godman, 1901 of Mnasinous Godman, 1900. Polites (Polites) coras (Cramer, 1775) is not a nomen dubium but a valid species. The following are species-level taxa (not subspecies or synonyms of taxa given in parenthesis): Lycaena pseudophlaeas (Lucas, 1866) and Lycaena hypophlaeas (Boisduval, 1852) (not Lycaena phlaeas (Linnaeus, 1761), Satyrium dryope (W. H. Edwards, 1870) (not Satyrium sylvinus (Boisduval, 1852)), Apodemia cleis (W. H. Edwards, 1882) (not Apodemia zela (Butler, 1870)), Epityches thyridiana (Haensch, 1909), comb. nov. (not Epityches ferra Haensch, 1909, comb. nov.), Argynnis bischoffii W. H. Edwards, 1870 (not Argynnis mormonia Boisduval, 1869), Argynnis leto Behr, 1862 (not Argynnis cybele (Fabricius, 1775)), Boloria myrina (Cramer, 1777) (not Boloria selene ([Denis & Schiffermüller], 1775)), Phyciodes jalapeno J. Scott, 1998 (not Phyciodes phaon (W. H. Edwards, 1864)), Phyciodes incognitus Gatrelle, 2004 and Phyciodes diminutor J. Scott, 1998 (not Phyciodes cocyta (Cramer, 1777)), Phyciodes orantain J. Scott, 1998 (not Phyciodes tharos (Drury, 1773)), Phyciodes anasazi J. Scott, 1994 (not Phyciodes batesii (Reakirt, [1866])), Cercyonis silvestris (W. H. Edwards, 1861) (not Cercyonis sthenele (Boisduval, 1852)), Paramacera allyni L. Miller, 1972 and Paramacera rubrosuffusa L. Miller, 1972 (not Paramacera xicaque (Reakirt, [1867])), Cissia cheneyorum (R. Chermock, 1949), Cissia pseudocleophes (L. Miller, 1976), and Cissia anabelae (L. Miller, 1976) (not Cissia rubricata (W. H. Edwards, 1871)), Tarsoctenus gaudialis (Hewitson, 1876) (not Tarsoctenus corytus (Cramer, 1777)), Nisoniades inca (Lindsey, 1925) (not Nisoniades mimas (Cramer, 1775), Xenophanes ruatanensis Godman & Salvin, 1895 (not Xenophanes tryxus (Stoll, 1780)), Lotongus shigeoi Treadaway & Nuyda, 1994, Lotongus balta Evans, 1949, Lotongus zalates (Mabille, 1893), and Lotongus taprobanus (Plötz, 1885) (not Lotongus calathus (Hewitson, 1876)), Oxynthes martius (Mabille, 1889) (not Oxynthes corusca (Herrich-Schäffer, 1869)), Notamblyscirtes durango J. Scott, 2017 (not Notamblyscirtes simius W. H. Edwards, 1881), Hedone praeceps Scudder, 1872, Hedone catilina (Plötz, 1886), and Hedone calla (Evans, 1955) (not Hedone vibex (Geyer, 1832)), Atalopedes huron (W. H. Edwards, 1863) (not Atalopedes campestris (Boisduval, 1852)), Papias microsema Godman, 1900 (not Mnasinous phaeomelas (Hübner, [1829]), comb. nov.), Papias unicolor (Hayward, 1938) and Papias monus Bell, 1942 (not Papias phainis Godman, 1900), Nastra leuconoides (Lindsey, 1925) (not Nastra leucone (Godman, 1900)), Nastra fusca (Grote & Robinson, 1867) (not Nastra lherminier (Latreille, [1824])), Zenis hemizona (Dyar, 1918) and Zenis janka Evans, 1955 (not Zenis jebus (Plötz, 1882)), Carystus (Argon) argus Möschler, 1879 (not Carystus (Argon) lota Hewitson, 1877), and Lycas devanes (Herrich-Schäffer, 1869) (not Lycas argentea (Hewitson, 1866)). Borbo impar ceramica (Plötz, 1886), comb. nov. is not a synonym of Pelopidas agna larika (Pagenstecher, 1884) but a valid subspecies. Parnassius smintheus behrii W. H. Edwards, 1870 and Cercyonis silvestris incognita J. Emmel, T. Emmel & Mattoon, 2012 are subspecies, not species. The following are junior subjective synonyms: Shijimiaeoides Beuret, 1958 of Glaucopsyche Scudder, 1872, Micropsyche Mattoni, 1981 of Turanana Bethune-Baker, 1916, Cyclyrius Butler, 1897 of Leptotes Scudder, 1876, Mesenopsis Godman & Salvin, 1886 of Xynias Hewitson, 1874, Carystus tetragraphus Mabille, 1891 of Lotongus calathus parthenope (Plötz, 1886), Parnara bipunctata Elwes & J. Edwards, 1897 of Borbo impar ceramica (Plötz, 1886), Hesperia peckius W. Kirby, 1837 of Polites (Polites) coras (Cramer, 1775), and Lerodea neamathla Skinner & R. Williams, 1923 of Nastra fusca (Grote & Robinson, 1867). The following transfers are proposed: of species between genera (i.e., revised genus-species combinations): Nervia niveostriga (Trimen, 1864) (not Kedestes Watson, 1893), Leona lota Evans, 1937 (not Lennia Grishin, 2022), Leona pruna (Evans, 1937) and Leona reali (Berger, 1962) (not Pteroteinon Watson, 1893), Mnasinous phaeomelas (Hübner, [1829]) (not Papias Godman, 1900), Saturnus jaguar (Steinhauser, 2008) (not Parphorus Godman, 1900), Parphorus harpe (Steinhauser, 2008) (not Saturnus Evans, 1955), Parphorus kadeni (Evans, 1955) (not Lento Evans, 1955), and Calpodes chocoensis (Salazar & Constantino, 2013) (not Megaleas Godman, 1901); of subspecies between species (i.e., revised species-subspecies combinations): Melitaea sterope W. H. Edwards, 1870 of Chlosyne palla (Boisduval, 1852) (not Chlosyne acastus (W. H. Edwards, 1874)) and Panoquina ocola distipuncta Johnson & Matusik, 1988 of Panoquina lucas (Fabricius, 1793); and junior subjective synonym transferred between species: Rhinthon zaba Strand, 1921 of Conga chydaea (A. Butler, 1877), not Cynea cynea (Hewitson, 1876), Pamphila stigma Skinner, 1896 of Hedone catilina (Plötz, 1886), not Hedone praeceps Scudder, 1872, and Pamphila ortygia Möschler, 1883 of Panoquina hecebolus (Scudder, 1872), not Panoquina ocola (W. H. Edwards, 1863). Proposed taxonomic changes result in additional revised species-subspecies combinations: Lycaena pseudophlaeas abbottii (Holland, 1892), Satyrium dryope putnami (Hy. Edwards, 1877), Satyrium dryope megapallidum Austin, 1998, Satyrium dryope itys (W. H. Edwards, 1882), Satyrium dryope desertorum (F. Grinnell, 1917), Argynnis bischoffi opis W. H. Edwards, 1874, Argynnis bischoffi washingtonia W. Barnes & McDunnough, 1913, Argynnis bischoffi erinna W. H. Edwards, 1883, Argynnis bischoffi kimimela Marrone, Spomer & J. Scott, 2008, Argynnis bischoffi eurynome W. H. Edwards, 1872, Argynnis bischoffi artonis W. H. Edwards, 1881, Argynnis bischoffi luski W. Barnes & McDunnough, 1913, Argynnis leto letona (dos Passos & Grey, 1945), Argynnis leto pugetensis (F. Chermock & Frechin, 1947), Argynnis leto eileenae (J. Emmel, T. Emmel & Mattoon, 1998), Boloria myrina nebraskensis (W. Holland, 1928), Boloria myrina sabulocollis Kohler, 1977, Boloria myrina tollandensis (W. Barnes & Benjamin, 1925), Boloria myrina albequina (W. Holland, 1928), Boloria myrina atrocostalis (Huard, 1927), Boloria myrina terraenovae (W. Holland, 1928), Phyciodes anasazi apsaalooke J. Scott, 1994, Polites coras surllano J. Scott, 2006, and Curva darienensis (Gaviria, Siewert, Mielke & Casagrande, 2018). Specimen curated as the holotype of Acidalia leto valesinoides-alba Reuss, [1926] is Argynnis leto letona (dos Passos & Grey, 1945) (not A. leto leto Behr, 1862) from USA: Utah, Provo. A synonymic list of available genus-group names for Lycaeninae [Leach],[1815] is given. Unless stated otherwise, all subgenera, species, subspecies and synonyms of mentioned genera and species are transferred with their parent taxa, and others remain as previously classified.

DNA barcodes reveal different speciation scenarios in the four North American Anthocharis Boisduval, Rambur, [Duménil] & Graslin, [1833] (Lepidoptera: Pieridae: Pierinae: Anthocharidini) species groups.

The mitochondrial DNA COI barcode segment sequenced from American Anthocharis specimens across their distribution ranges partitions them into four well-separated species groups and reveals different levels of differentiation within these groups. The lanceolata group experienced the deepest divergence. About 2.7% barcode difference separates the two species: A. lanceolata Lucas, 1852 including A. lanceolata australis (F. Grinnell, 1908), from A. desertolimbus J. Emmel, T. Emmel & Mattoon, 1998. The sara group consists of three species distinctly defined by more than 2% sequence divergence: A. sara Lucas, 1852, A. julia W. H. Edwards, 1872, and A. thoosa (Scudder, 1878). Our treatment is fully consistent with morphological evidence largely based on the characters of fifth instar larvae and pupal cone curvature (Stout, 2005, 2018). In barcodes, it is not possible to see evidence of introgression or hybridization between the three species, and identification by morphology of immature stages always agrees with DNA barcode identification. Interestingly, A. thoosa exhibited the largest intraspecific divergence in DNA barcodes, and several of its metapopulations are identifiable by haplotypes. The cethura group is characterized by the smallest divergence and is best considered as a single species variable in expression of yellow coloration: A cethura C. Felder & R. Felder, 1865. Notably, the most sexually dimorphic subspecies A. cethura morrisoni W. H. Edwards, 1881 is the most distinct by the barcodes. Finally, the midea group barcodes do not always separate A. midea (Hübner, [1809]) and A. limonea (A. Butler, 1871) and we observe gradual accumulation of differences from north (northeastern USA) to south (Hidalgo, Mexico). This barcode gradient suggests a recent origin of the two midea group species and provides another example of vicariant sister species well defined by morphology, ecology and geography, but not necessarily by DNA barcodes.

A DNA barcode library for the butterflies of North America.

Although the butterflies of North America have received considerable taxonomic attention, overlooked species and instances of hybridization continue to be revealed. The present study assembles a DNA barcode reference library for this fauna to identify groups whose patterns of sequence variation suggest the need for further taxonomic study. Based on 14,626 records from 814 species, DNA barcodes were obtained for 96% of the fauna. The maximum intraspecific distance averaged 1/4 the minimum distance to the nearest neighbor, producing a barcode gap in 76% of the species. Most species (80%) were monophyletic, the others were para- or polyphyletic. Although 15% of currently recognized species shared barcodes, the incidence of such taxa was far higher in regions exposed to Pleistocene glaciations than in those that were ice-free. Nearly 10% of species displayed high intraspecific variation (>2.5%), suggesting the need for further investigation to assess potential cryptic diversity. Aside from aiding the identification of all life stages of North American butterflies, the reference library has provided new perspectives on the incidence of both cryptic and potentially over-split species, setting the stage for future studies that can further explore the evolutionary dynamics of this group.

Genomics-guided refinement of butterfly taxonomy.

Continuing with comparative genomic exploration of worldwide butterfly fauna, we use all protein-coding genes as they are retrieved from the whole genome shotgun sequences for phylogeny construction. Analysis of these genome-scale phylogenies projected onto the taxonomic classification and the knowledge about butterfly phenotypes suggests further refinements of butterfly taxonomy that are presented here. As a general rule, we assign most prominent clades of similar genetic differentiation to the same taxonomic rank, and use criteria based on relative population diversification and the extent of gene exchange for species delimitation. As a result, 7 tribes, 4 subtribes, 14 genera, and 9 subgenera are proposed as new, i.e., in subfamily Pierinae Swainson, 1820: Calopierini Grishin, trib. n. (type genus Calopieris Aurivillius, 1898); in subfamily Riodininae Grote, 1895: Callistiumini Grishin, trib. n. (type genus Callistium Stichel, 1911); in subfamily Nymphalinae Rafinesque, 1815: Pycinini Grishin, trib. n. (type genus Pycina Doubleday 1849), Rhinopalpini Grishin, trib. n. (type genus Rhinopalpa C. & R. Felder 1860), Kallimoidini Grishin, trib. n. (type genus Kallimoides Shirôzu & Nakanishi 1984), Vanessulini Grishin, trib. n. (type genus Vanessula Dewitz 1887), and Doleschalliaini Grishin, trib. n. (type genus Doleschallia C. & R. Felder 1860); in tribe Mesosemiini Bates, 1859: Eunogyrina Grishin, subtrib. n. (type genus Eunogyra Westwood, 1851); in tribe Satyrini Boisduval, 1833: Callerebiina Grishin, subtrib. n. (type genus Callerebia Butler, 1867), Gyrocheilina Grishin, subtrib. n. (type genus Gyrocheilus Butler, 1867), and Calistina Grishin, subtrib. n. (type genus Calisto Hübner, [1823]); in subfamily Euselasiinae Kirby, 1871: Pelolasia Grishin, gen. n. (type species Eurygona pelor Hewitson, [1853]), Myselasia Grishin, gen. n. (type species Eurygona mys Herrich-Schäffer, [1853]), Eurylasia Grishin, gen. n. (type species Eurygona euryone Hewitson, 1856), Maculasia Grishin, gen. n. (type species Euselasia albomaculiga Callaghan, 1999), and Eugelasia Grishin, gen. n. (type species Eurygona eugeon Hewitson, 1856); in subtribe Mesosemiina Bates, 1859: Ectosemia Grishin, gen. n. (type species Papilio eumene Cramer, 1776) and Endosemia Grishin, gen. n. (type species Papilio ulrica Cramer, 1777); in tribe Symmachiini Reuter, 1896: Tigria Grishin, gen. n. (type species Mesene xypete Hewitson, 1870) and Asymma Grishin, gen. n. (type species Symmachia virgatula Stichel, 1910); in tribe Riodinini Grote, 1895: Putridivora Grishin, gen. n. (type species Charis argyrea Bates, 1868), Chadia Grishin, gen. n. (type species Charis cadytis Hewitson, 1866), Inkana Grishin, gen. n. (type species Charis incoides Schaus, 1902), and Oco Grishin, gen. n. (type species Symmachia ocellata Hewitson, 1867); in subtribe Zabuellina Seraphim, Freitas & Kaminski, 2018: Teenie Grishin, gen. n. (type species Calydna tinea Bates, 1868); Boreographium Grishin, subgen. n. (type species Papilio marcellus Cramer, 1777, parent genus Eurytides Hübner, [1821]), Esperourus Grishin, subgen. n. (type species Papilio esperanza Beutelspacher, 1975, parent genus Pterourus Scopoli, 1777), Hyppasonia Grishin, subgen. n. (type species Papilio hyppason Cramer, 1775, parent genus Heraclides Hübner, [1819]), Sisymbria Grishin, subgen. n. (type species Pieris sisymbrii Boisduval, 1852, parent genus Pontia [Fabricius], 1807), Greenie Grishin, subgen. n. (type species Thecla sheridonii [sic] Edwards, 1877, parent genus Callophrys Billberg, 1820), Magda Grishin, subgen. n. (type species Erebia magdalena Strecker, 1880, parent genus Erebia Dalman, 1816), and in genus Eresia Boisduval, 1836: Notilia Grishin, subgen. n. (type species Eresia orthia Hewitson, 1864), Levinata Grishin, subgen. n. (type species Eresia levina Hewitson, 1872), and Ithra Grishin, subgen. n. (type species Phyciodes ithra Kirby, 1900). Furthermore, we resurrect 6 genera, change the rank of 36 currently used genera to subgenus, synonymize 3 subtribes, 42 genera or subgenera, assign 3 genera to tribes and subtribes, and transfer 34 additional species to genera different from those these taxa are presently assigned to, present evidence to support 7 taxa as species instead of subspecies, and 1 taxon as a subspecies instead of species. Namely, the following taxa are valid genera: Terias Swainson, 1821 (not in Eurema Hübner, [1819]), Erythia Hübner, [1819] and Marmessus Hübner, [1819] (not in Euselasia Hübner, [1819]), Eucorna Strand, 1932 (not in Voltinia Stichel, 1910), Cremna Doubleday, 1847 (not in Napaea Hübner, [1819]), and Hallonympha Penz & DeVries, 2006 (not in Zabuella Stichel, 1911). The following taxa are best treated as subgenera: Zegris Boisduval, 1836 of Anthocharis Boisduval, Rambur, [Duménil] & Graslin, [1833]; Baltia Moore, 1878 and Pontieuchloia Verity, 1929 of Pontia [Fabricius], 1807; Phrissura Butler, 1870 of Appias Hübner, [1819]; Saletara Distant, 1885 of Catophaga Hübner, 1819; Leodonta Butler, 1870 of Pereute Herrich-Schäffer, 1867; Takashia M. Okano & T. Okano, 1985 of Polycaena Staudinger, 1886; Corrachia Schaus, 1913 of Styx Staudinger, 1876; Ionotus Hall, 2005 and Voltinia Stichel, 1910 of Cremna Doubleday, 1847; Hermathena Hewitson, 1874 of Ithomiola C. & R. Felder, 1865; Lucillella Strand, 1932 of Esthemopsis C. & R. Felder, 1865; Mesenopsis Godman & Salvin, 1886 and Xenandra C. & R. Felder, 1865 of Symmachia Hübner, [1819]; Pirascca J. Hall & Willmott, 1996 of Pterographium Stichel, 1910; Imelda Hewitson, 1870 of Echenais Hübner, [1819]; Calicosama J. Hall & Harvey, 2001 of Behemothia Hall, 2000; Polygrapha Staudinger, 1887 and Fountainea Rydon, 1971 of Anaea Hübner, [1819]; Siderone Hübner, [1823] and Phantos Dias, 2018 of Zaretis Hübner, [1819]; Harsiesis Fruhstorfer, 1911 of Platypthima Rothschild & Jordan, 1905; Vila Kirby, 1871 of Biblis Fabricius, 1807; Diaethria Billberg, 1820 and Perisama Doubleday, 1849 of Callicore Hübner, [1819]; Antigonis C. Felder, 1861 of Haematera Doubleday, 1849; Asterope Hübner, [1819], Nica Hübner, [1826], Peria Kirby, 1871, and Callicorina Smart, 1976 of Temenis Hübner, [1819]; Anthanassa Scudder, 1875, Castilia Higgins, 1981, Telenassa Higgins, 1981, Dagon Higgins, 1981, and Janatella Higgins, 1981 of Eresia Boisduval, 1836; and Wallengrenia Berg, 1897 of Polites Scudder, 1872. The following taxa are junior subjective synonyms: Maniolina Grote, 1897 of Erebiina Tutt, 1896; Melanargiina Wheeler, 1903 of Satyrina Boisduval, 1833; Phyciodina Higgins, 1981 of Melitaeina Herrich-Schäffer, 1843; Cunizza Grote, 1900 of Hesperocharis C. Felder, 1862; Reliquia Ackery, 1975 of Pontia [Fabricius], 1807; Tatochila A. Butler, 1870, Piercolias Staudinger, 1894, Hypsochila Ureta, 1955, Theochila W. D. Field, 1958, Pierphulia W. D. Field, 1958, and Infraphulia W. D. Field, 1958 of Phulia Herrich-Schäffer, 1867; Mesapia Gray, 1856 of Aporia Hübner, [1819]; Catasticta Butler, 1870 of Archonias Hübner, 1827; Sandia Clench & P. Ehrlich, 1960 andXamia Clench, 1961 of Incisalia Scudder, 1872; Hades Westwood, 1851 of Methone Doubleday, 1847; Semomesia Westwood, 1851, Mesophthalma Westwood, 1851, Perophthalma Westwood, 1851 and Leucochimona Stichel, 1909 of Mesosemia Hübner, [1819], Xynias Hewitson, 1874 of Mesenopsis Godman & Salvin, 1886; Stichelia J. Zikán, 1949 of Symmachia Hübner, [1819]; Chimastrum Godman & Salvin, 1886 of Mesene Doubleday, 1847; Alethea Nielsen & Salazar, [2018] of Pirascca J. Hall & Willmott, 1996; Panaropsis J. Hall, 2002 of Pterographium Stichel, 1910; Comphotis Stichel, 1910 of Phaenochitonia Stichel, 1910; Colaciticus Stichel, 1910 of Baeotis Hübner, [1819]; Nahida Kirby, 1871 of Ithomeis Bates, 1862; Machaya Hall & Willmott, 1995 of Pachythone Bates, 1868; Percnodaimon Butler, 1876 and Erebiola Fereday, 1879 of Argyrophenga Doubleday, 1845; Hestinalis Bryk, 1938 of Mimathyma Moore, 1896; Catacore Dillon, 1948 of Diaethria Billberg, 1820; Mesotaenia Kirby, 1871 and Orophila Staudinger, 1886 of Perisama Doubleday, 1849; Paulogramma Dillon, 1948 of Catagramma Boisduval, 1836; Panacea Godman & Salvin, 1883 of Batesia C. Felder & R. Felder, 1862; Napeocles Bates, 1864 of Siproeta Hübner, [1823]; Texola Higgins, 1959 and Dymasia Higgins, 1960 of Microtia H. Bates, 1864; Tisona Higgins, 1981 of Ortilia Higgins, 1981; Abananote Potts, 1943 and Altinote Potts, 1943 of Actinote Hübner, [1819]; Episcada Godman & Salvin, 1879 of Ceratinia Hübner, 1816; and Appia Evans, 1955 of Pompeius Evans, 1955. The following genera are placed in taxonomic hierarchy: Prestonia Schaus, 1920 belongs to Euremini Grote, 1898; Petrocerus Callaghan, 1979 belongs to Theopina Clench, 1955; and Paralasa Moore, 1893 belongs to Ypthimina Reuter, 1896. The following taxa are distinct species rather than subspecies (of species shown in parenthesis): Pyrisitia westwoodii (Boisduval, 1836) (not Pyrisitia dina (Poey, 1832)), Biblis aganisa Boisduval, 1836 (not Biblis hyperia (Cramer, 1779)), Phystis variegata (Röber, 1913) and Phystis pratti (A. Hall, 1935) (not Phystis simois (Hewitson, 1864)), Phocides batabano (Lucas, 1857) and Phocides bicolora (Boddaert, 1783) (not Phocides pigmalion (Cramer, 1779)), Lobotractus mysie (Dyar, 1904) (not Lobotractus valeriana (Plötz, 1881)). Nahida coenoides (Hewitson, 1870) is conspecific with Ithomeis aurantiaca H. Bates, 1862. Additional new and revised combinations are: Teriocolias deva (E. Doubleday, 1847), Teriocolias reticulata (A. Butler, 1871), Hesperocharis leucothea (Molina, 1782), Methone euploea (Hewitson, [1855]), Methone eucerus (Hewitson, 1872), Methone hypophaea (Godman & Salvin, 1878), Methone eubule (R. Felder, 1869), Methone onorata (Hewitson, 1869), Methone authe (Godman, 1903), Methone dolichos (Staudinger, [1887]), Methone baucis (Stichel, 1919), Methone eucrates (Hewitson, 1872), Napaeadanforthi A. Warren & Opler, 1999, Napaea dramba (J. Hall, Robbins & Harvey, 2004), Napaea sanarita (Schaus, 1902), Napaea agroeca Stichel, 1910, Napaea tumbesia J. Hall & Lamas, 2001, Napaea umbra (Boisduval, 1870), Napaea phryxe (C. & R. Felder, 1865), Napaea cebrenia (Hewitson, [1873]), Napaea loxicha (R.G. Maza & J. Maza, 2016), Napaea maya (J. Maza & Lamas, 2016), Napaea necaxa (R.G. Maza & J. Maza, 2018), Napaea totonaca (R.G. Maza & J. Maza, 2016), Mesene aeolia (Bates, 1868), Pterographium hypochloris (Bates, 1868), Phaenochitonia florus (Fabricius, 1793), Ourocnemis carausius (Westwood, 1851), Ourocnemis principalis (Hopffer, 1874), Ourocnemis renaldus (Stoll, 1790), and Ourocnemis aerosus (Stichel, 1924), Hallonympha maculosa (Bates, 1868), Exoplisia aphanis (Stichel, 1910), Phystis fontus (A. Hall, 1928), Phocides batabano okeechobee (Worthington, 1881), and Phocides batabano batabanoides (W. Holland, 1902). Finally, we confirm the combination Zabuella castanea (Prittwitz, 1865) and find Pyrgus centaureae dzekh Gorbunov, 2007 as a new subspecies for North America.

Genomic evidence suggests further changes of butterfly names.

Further genomic sequencing of butterflies by our research group expanding the coverage of species and specimens from different localities, coupled with genome-scale phylogenetic analysis and complemented by phenotypic considerations, suggests a number of changes to the names of butterflies, mostly those recorded from the United States and Canada. Here, we present evidence to support these changes. The changes are intended to make butterfly classification more internally consistent at the genus, subgenus and species levels. I.e., considering all available evidence, we attempt to assign similar taxonomic ranks to the clades of comparable genetic differentiation, which on average is correlated with the age of phylogenetic groups estimated from trees. For species, we use criteria devised by genomic analysis of the genetic differentiation across suture zones and comparison of sympatric populations of closely related species. As a result, we resurrect 4 genera and 1 subgenus from subgeneric status or synonymy, change the rank of 8 currently used genera to subgenus, synonymize 7 genus-group names, summarize evidence to support 19 taxa as species instead of subspecies and 1 taxon as subspecies instead of species, along with a number of additional changes. One new genus and one new subspecies are described. Namely, the following taxa are treated as genera Tharsalea Scudder, 1876, Helleia Verity, 1943, Apangea Zhdanko, 1995, and Boldenaria Zhdanko, 1995. Tetracharis Grote, 1898 is a valid subgenus (not a synonym of Anthocharis Boisduval, Rambur, [Duménil] & Graslin, [1833]) that consists of Anthocharis cethura C. Felder & R. Felder, 1865 (Müller, 1764), Anthocharis midea (Hübner, [1809]), and Anthocharis limonea (A. Butler, 1871). The following are subgenera: Speyeria Scudder, 1872 of Argynnis Fabricius, 1807; Aglais Dalman, 1816 and Polygonia Hübner, [1819] of Nymphalis Kluk, 1780; Palaeonympha Butler, 1871 of Megisto Hübner, [1819]; Hyponephele Muschamp, 1915 of Cercyonis Scudder, 1875; Pyronia Hübner, [1819] and Aphantopus Wallengren, 1853 of Maniola Schrank, 1801 and Pseudonymphidia Callaghan, 1985 of Pachythone. Lafron Grishin, gen. n. (type species Papilio orus Stoll, [1780], parent subfamily Lycaeninae [Leach], [1815]) is described. Dipsas japonica Murray, 1875 is fixed as the type species of Neozephyrus Sibatani & Ito, 1942. The following taxa are junior subjective synonyms: Falcapica Klots, 1930 of Tetracharis Grote, 1898; Habrodais Scudder, 1876, Favonius Sibatani & Ito, 1942, Neozephyrus Sibatani & Ito, 1942, Quercusia Verity, 1943, Chrysozephyrus Shirôzu & Yamamoto, 1956, and Sibataniozephyrus Inomata, 1986 of Hypaurotis Scudder, 1876; Plesioarida Trujano & García, 2018 of Roeberella Strand, 1932; Papilio temenes Godart, 1819 (lectotype designated herein) of Heraclides aristodemus (Esper, 1794), Speyeria hydaspe conquista dos Passos & Grey, 1945 of Argynnis hesperis tetonia (dos Passos & Grey, 1945), and Erycides imbreus Plötz, 1879 of Phocides polybius polybius (Fabricius, 1793). The following are revised genus-species combinations: Pachythone lencates (Hewitson, 1875) Pachythone flocculus (Brévignon & Gallard, 1993), Pachythone floccus (Brévignon, 2013), Pachythone heberti (P. Jauffret & J. Jauffret, 2007), Pachythone marajoara (P. Jauffret & J. Jauffret, 2007) and Cissia cleophes (Godman & Salvin, 1889). The following species are transferred between subgenera: Anthocharis lanceolata Lucas, 1852 belongs to Anthocharis Boisduval, Rambur, [Duménil] & Graslin, [1833] instead of Paramidea Kuznetsov, 1929 and Danaus eresimus (Cramer, 1777) belongs to Danaus Kluk, 1780, and not to Anosia Hübner, 1816. The following taxa are distinct species rather than subspecies (of species shown in parenthesis): Heraclides ponceana (Schaus, 1911) (not Heraclides aristodemus (Esper, 1794)), Colias elis Strecker, 1885 (not Colias meadii W. H. Edwards, 1871), Argynnis irene Boisduval, 1869 and Argynnis nausicaa W. H. Edwards, 1874 (not Argynnis hesperis W. H. Edwards, 1864), Coenonympha california Westwood, [1851] (not Coenonympha tullia (Müller, 1764)), Dione incarnata N. Riley, 1926 (not Dione vanillae (Linnaeus, 1758)), Chlosyne coronado (M. Smith & Brock, 1988) (not Chlosyne fulvia (W. H. Edwards, 1879)), Chlosyne chinatiensis (Tinkham, 1944) (not Chlosyne theona (Ménétriés, 1855)), Phocides lilea (Reakirt, [1867]) (not Phocides polybius (Fabricius, 1793)), Cecropterus nevada (Scudder, 1872) and Cecropterus dobra (Evans, 1952) (not Cecropterus mexicana (Herrich-Schäffer, 1869)), Telegonus anausis Godman & Salvin, 1896, (not Telegonus anaphus (Cramer, 1777)), Epargyreus huachuca Dixon, 1955 (not Epargyreus clarus (Cramer, 1775)), Nisoniades bromias (Godman & Salvin, 1894) (not Nisoniades rubescens (Möschler, 1877)), Pholisora crestar J. Scott & Davenport, 2017 (not Pholisora catullus (Fabricius, 1793)), Carterocephalus mandan (W. H. Edwards, 1863) and Carterocephalus skada (W. H. Edwards, 1870) (not Carterocephalus palaemon (Pallas, 1771)), Amblyscirtes arizonae H. Freeman, 1993 (not Amblyscirtes elissa Godman, 1900), and Megathymus violae D. Stallings & Turner, 1956 (not Megathymus ursus Poling, 1902). Resulting from these changes, the following are revised species-subspecies combinations: Heraclides ponceana bjorndalae (Clench, 1979), Heraclides ponceana majasi L. Miller, 1987, Argynnis irene dodgei Gunder, 1931, Argynnis irene cottlei J. A. Comstock, 1925, Argynnis irene hanseni (J. Emmel, T. Emmel & Mattoon, 1998), Argynnis nausicaa elko (Austin, 1984), Argynnis nausicaa greyi (Moeck, 1950), Argynnis nausicaa viola (dos Passos & Grey, 1945), Argynnis nausicaa tetonia (dos Passos & Grey, 1945), Argynnis nausicaa chitone W. H. Edwards, 1879, Argynnis nausicaa schellbachi (Garth, 1949), Argynnis nausicaa electa W. H. Edwards, 1878, Argynnis nausicaa dorothea (Moeck, 1947), and Argynnis nausicaa capitanensis (R. Holland, 1988), Argynnis zerene atossa W. H. Edwards, 1890, Dione incarnata nigrior (Michener, 1942), Chlosyne coronado pariaensis (M. Smith & Brock, 1988), Cecropterus nevada aemilea (Skinner, 1893), Cecropterus nevada blanca (J. Scott, 1981), Telegonus anausis annetta (Evans, 1952), Telegonus anausis anoma (Evans, 1952), Telegonus anausis aniza (Evans, 1952), Epargyreus huachuca profugus Austin, 1998, Carterocephalus mandan mesapano (Scudder, 1868) and Carterocephalus skada magnus Mattoon & Tilden, 1998. American Coenonympha subspecies placed under C. tullia other than Coenonympha tullia kodiak W. H. Edwards, 1869, Coenonympha tullia mixturata Alpheraky, 1897 and Coenonympha tullia yukonensis W. Holland, 1900 belong to C. california. Heraclides ponceana latefasciatus Grishin, ssp. n. is described from Cuba. Argynnis coronis carolae dos Passos & Grey, 1942 is considered a subspecies-level taxon. Unless stated otherwise, all subgenera, species, subspecies and synonyms of mentioned genera and species are transferred together with their parent taxa, and others remain as previously classified.

Changes to North American butterfly names.

We obtained and analyzed whole genome shotgun sequences of all 845 species of butterflies recorded from Canada and the United States. Genome-scale phylogenetic trees constructed from the data reveal several non-monophyletic genera and suggest improved classification of species included in these genera. Here, these changes are formalized and 2 subgenera are described: Amblyteria Grishin, subgen. n. (type species Goniloba exoteria Herrich-Schäffer, 1869, parent genus Amblyscirtes Scudder, 1872), and Coa Grishin, subgen. n. (type species Hesperia baracoa Lucas, 1857, parent genus Polites Scudder, 1872). Furthermore, we resurrect 3 genera and 2 subgenera from synonymy, change the rank of 6 currently used genera to subgenus, synonymize 2 genera, transfer 3 (2 resurrected) subgenera and 11 additional species to different genera than those these taxa were assigned to, and raise one name from synonym to species rank. Namely, Hedone Scudder, 1872 and Limochores Scudder, 1872 are valid genera and not synonyms of Polites Scudder, 1872; Pendantus K. Johnson & Kroenlein, 1993 is a valid genus and not a synonym of Electrostrymon Clench, 1961; and Sphaenogona Butler, 1870 and Lucidia Lacordaire, 1833 are valid subgenera of Abaeis Hübner, [1819] (new placement) and not synonyms of Eurema Hübner, [1819]. The following taxa are best treated as subgenera: Mimoides Brown, 1991 of Eurytides Hübner, [1821] (sensu lato); Philotiella Mattoni, [1978] of Euphilotes Mattoni, [1978]; Neominois Scudder, 1875 of Oeneis Hübner, [1819]; Agraulis Boisduval & Le Conte, [1835] of Dione Hübner, [1819]; Copaeodes Speyer, 1877 of Oarisma Scudder, 1872; and Problema Skinner & R. Williams, 1924 of Atrytone Scudder, 1872. Phaeostrymon Clench, 1961 and Saliana Evans, 1955 are junior subjective synonyms of Satyrium Scudder, 1876 and Calpodes Hübner, [1819], respectively. The entire subgenus Erynnides Burns, 1964 is transferred from Erynnis Schrank, 1801 to Gesta Evans, 1953. New genus-species combinations resulting from transfer of species between genera are: Nastra perigenes (Godman, 1900) (not Vidius Evans, 1955); Troyus fantasos (Cramer, 1780), Troyus onaca (Evans, 1955), Troyus aurelius (Plötz, 1882), Troyus marcus (Fabricius, 1787), Troyus diversa (Herrich-Schäffer, 1869), and Troyus drova (Evans, 1955) (not Vettius Godman, 1901); Oligoria percosius (Godman, 1900), Oligoria rindgei (H. Freeman, 1969), Oligoria lucifer (Hübner, [1831]), and Oligoria mustea (H. Freeman, 1979) (not Decinea Evans, 1955). Urbanus alva Evans, 1952 is a valid species and not a synonym of Urbanus belli (Hayward, 1935), new status.

LepNet: The Lepidoptera of North America Network.

The Lepidoptera of North America Network, or LepNet, is a digitization effort recently launched to mobilize biodiversity data from 3 million specimens of butterflies and moths in United States natural history collections (http://www.lep-net.org/). LepNet was initially conceived as a North American effort but the project seeks collaborations with museums and other organizations worldwide. The overall goal is to transform Lepidoptera specimen data into readily available digital formats to foster global research in taxonomy, ecology and evolutionary biology.

When COI barcodes deceive: complete genomes reveal introgression in hairstreaks.

Two species of hairstreak butterflies from the genus Calycopis are known in the United States: C. cecrops and C. isobeon Analysis of mitochondrial COI barcodes of Calycopis revealed cecrops-like specimens from the eastern US with atypical barcodes that were 2.6% different from either USA species, but similar to Central American Calycopis species. To address the possibility that the specimens with atypical barcodes represent an undescribed cryptic species, we sequenced complete genomes of 27 Calycopis specimens of four species: C. cecrops, C. isobeon, C. quintana and C. bactra Some of these specimens were collected up to 60 years ago and preserved dry in museum collections, but nonetheless produced genomes as complete as fresh samples. Phylogenetic trees reconstructed using the whole mitochondrial and nuclear genomes were incongruent. While USA Calycopis with atypical barcodes grouped with Central American species C. quintana by mitochondria, nuclear genome trees placed them within typical USA C. cecrops in agreement with morphology, suggesting mitochondrial introgression. Nuclear genomes also show introgression, especially between C. cecrops and C. isobeon About 2.3% of each C. cecrops genome has probably (p-value < 0.01, FDR < 0.1) introgressed from C. isobeon and about 3.4% of each C. isobeon genome may have come from C. cecrops. The introgressed regions are enriched in genes encoding transmembrane proteins, mitochondria-targeting proteins and components of the larval cuticle. This study provides the first example of mitochondrial introgression in Lepidoptera supported by complete genome sequencing. Our results caution about relying solely on COI barcodes and mitochondrial DNA for species identification or discovery.

Genetic evaluation of the evolutionary distinctness of a federally endangered butterfly, Lange's Metalmark.

BACKGROUND: The Mormon Metalmark (Apodemia mormo) species complex occurs as isolated and phenotypically variable colonies in dryland areas across western North America. Lange's Metalmark, A. m. langei, one of the 17 subspecies taxonomically recognized in the complex, is federally listed under the U.S. Endangered Species Act of 1973. Metalmark taxa have traditionally been described based on phenotypic and ecological characteristics, and it is unknown how well this nomenclature reflects their genetic and evolutionary distinctiveness. Genetic variation in six microsatellite loci and mitochondrial cytochrome oxidase subunit I sequence was used to assess the population structure of the A. mormo species complex across 69 localities, and to evaluate A. m. langei's qualifications as an Evolutionarily Significant Unit. RESULTS: We discovered substantial genetic divergence within the species complex, especially across the Continental Divide, with population genetic structure corresponding more closely with geographic proximity and local isolation than with taxonomic divisions originally based on wing color and pattern characters. Lange's Metalmark was as genetically divergent as several other locally isolated populations in California, and even the unique phenotype that warranted subspecific and conservation status is reminiscent of the morphological variation found in some other populations. CONCLUSIONS: This study is the first genetic treatment of the A. mormo complex across western North America and potentially provides a foundation for reassessing the taxonomy of the group. Furthermore, these results illustrate the utility of molecular markers to aid in demarcation of biological units below the species level. From a conservation point of view, Apodemia mormo langei's diagnostic taxonomic characteristics may, by themselves, not support its evolutionary significance, which has implications for its formal listing as an Endangered Species.