Background: The goal of DNA barcoding is to develop a species-specific sequence library for all eukaryotes. A 650 bp fragment of the cytochrome c oxidase 1 (CO1) gene has been used successfully for species-level identification in several animal groups. It may be difficult in practice, however, to retrieve a 650 bp fragment from archival specimens, (because of DNA degradation) or from environmental samples (where universal primers are needed).

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This study reports DNA barcodes for more than 1300 Lepidoptera species from the eastern half of North America, establishing that 99.3 per cent of these species possess diagnostic barcode sequences. Intraspecific divergences averaged just 0.43 per cent among this assemblage, but most values were lower. The mean was elevated by deep barcode divergences (greater than 2%) in 5.1 per cent of the species, often involving the sympatric occurrence of two barcode clusters. A few of these cases have been analysed in detail, revealing species overlooked by the current taxonomic system. This study also provided a large-scale test of the extent of regional divergence in barcode sequences, indicating that geographical differentiation in the Lepidoptera of eastern North America is small, even when comparisons involve populations as much as 2800 km apart. The present results affirm that a highly effective system for the identification of Lepidoptera in this region can be built with few records per species because of the limited intra-specific variation. As most terrestrial and marine taxa are likely to possess a similar pattern of population structure, an effective DNA-based identification system can be developed with modest effort.

Many cold adapted species occur in both montane settings and in the subarctic. Their disjunct distributions create taxonomic complexity because there is no standardized method to establish whether their allopatric populations represent single or different species. This study employs DNA barcoding to gain new perspectives on the levels and patterns of sequence divergence among populations of 122 arctic-alpine species of Lepidoptera from the Alps, Fennoscandia and North America. It reveals intraspecific variability in the barcode region ranging from 0.00–10.08%. Eleven supposedly different species pairs or groups show close genetic similarity, suggesting possible synonymy in many cases. However, a total of 33 species show evidence of cryptic diversity as evidenced by the presence of lineages with over 2% maximum barcode divergence in Europe, in North America or between the two continents. Our study also reveals cases where taxonomic names have been used inconsistently between regions and exposes misidentifications. Overall, DNA barcodes have great potential to both increase taxonomic resolution and to make decisions concerning the taxonomic status of allopatric populations more objective.

DNA barcoding is an effective approach for species identification and for discovery of new and/or cryptic species. Sanger sequencing technology is the method of choice for obtaining standard 650 bp cytochrome c oxidase subunit I (COI) barcodes. However, DNA degradation/fragmentation makes it difficult to obtain a full-length barcode from old specimens. Mini-barcodes of 130 bp from the standard barcode region have been shown to be effective for accurate identification in many animal groups and may be readily obtained from museum samples. Here we demonstrate the application of an alternative sequencing technology, the four-enzymes single-specimen pyrosequencing, in rapid, cost-effective mini-barcode analysis. We were able to generate sequences of up to 100 bp from mini-barcode fragments of COI in 135 fresh and 50 old Lepidoptera specimens (ranging from 53–97 year-old). The sequences obtained using pyrosequencing were of high quality and we were able to robustly match all the tested pyro-sequenced samples to their respective Sanger-sequenced standard barcode sequences, where available. Simplicity of the protocol and instrumentation coupled with higher speed and lower cost per sequence than Sanger sequencing makes this approach potentially useful in efforts to link standard barcode sequences from unidentified specimens to known museum specimens with only short DNA fragments.

Genera and previously described species of Nearctic Scythrididae are revised for the first time, based on the study of adult structures. About 90 percent of the Nearctic fauna known in collections consists of undescribed species. The supraspecific taxa treated in this work encompass less than half of the Nearctic species diversity. Only six new species are described, all within the largest and structurally most diverse genus. The status of all nominal species is revised. Valid species are redescribed and their features illustrated. General problems in the systematics of the Scythrididae are discussed. A description of adult features of the family Scythrididae is providad. Extra-limital genera are briefly reviewed. A key to the Nearctic genera and informal supraspecific lineages is provided.Six genera, including three new, are treated: Areniscythris Powell, 1976, Arotrura Walsingham, 1888, Asymmetrura gen. nov., Neoscythris gen. nov., Rhamphura gen. nov., and Scythris s. str. Hübner, [1825]. Areniscythris includes a single described species, Areniscythris brachypteris Powell, but is defined more broadly to account for a number of undescribed species. Arotrura is divided into nine informal species groups with the following included species: Arotrura atascosa sp. nov., Arotrura balli sp. nov., Arotrura divaricata (Braun) comb, nov., Arotrura eburnea Walsingham, Arotrura formidabilis sp. nov., Arotrura hymenata sp. nov., Arotrura longissima sp. nov., Arotrura oxyplecta (Meyrick) comb, nov., Arotrura powelli sp. nov., and Arotrura sponsella (Busck) comb. nov. Asymmetrura includes: Asymmetrura albilineata (Walsingham) comb. nov., Asymmetrura graminivorella (Braun) comb. nov., Asymmetrura impositella (Zeller) comb. nov. and type species, Asymmetrura matutella (Clemens) comb, nov., Asymmetrura reducta (Braun) comb, nov., and Asymmetrura scintillifera (Braun) comb. nov. Neoscythris includes: Neoscythris confinis (Braun) comb, nov., Neoscythris euthia (Walsingham) comb. nov., Neoscythris fissirostris (Meyrick) comb. nov. and type species, and Neoscythris planipenella (Chambers) comb. nov. Rhamphura includes: Rhamphura altisierrae (Keifer) comb, nov., Rhamphura ochristriata (Walsingham) comb. nov. and type species, Rhamphura perspicillella (Walsingham) comb. nov., Rhamphura suffusa (Walsingham) comb. nov., and the extra-limital Rhamphura immunis (Meyrick) comb. nov. from Peru. Scythris s. str. includes: Scythris immaculatella (Chambers) rev. stat., Scythris limbella (Fabricius), Scythris mixaula Meyrick, Scythris trivinctella (Zeller), and Scythris ypsilon Braun. A further eight species are phylogenetically distinct from Scythris s. str. but provisionally are only assigned to five informal monophyletic lineages until their cladistic relationships are more firmly established. These are: the Scythris basilaris lineage including Scythris basilaris (Zeller), Scythris eboracensis (Zeller), and Scythris fuscicomella (Clemens); the Scythris interrupta lineage including Scythris interrupta Braun; the Scythris inspersella lineage including Scythris inspersella (Hübner) and Scythris noricella (Zeller); the Scythris anthracina lineage including Scythris anthracina Braun; and the Scythris charon lineage including Scythris charon Meyrick. Three species are incertae sedis: Scythris inornatella (Chambers) comb, nov., Scythrispilosella (Zeller), and Scythris piratica Meyrick.Coleophora albacostella Chambers and Coleophora inornatella Chambers are transferred from the Coleophoridae. Scythris arizoniella (Kearfott) is transferred to the Coleophoridae [Coleophora arizoniella (Kearfott) comb. nov.].The following new synonymy is proposed: Colinita Busck, 1907 = Arotrura Walsingham, 1888; Gelechia aterrimella Walker, 1864 and Scythris epilobiella McDunnough, 1942 = Scythris inspersella [Hübner, (1817)]; Scythris magnatella Busck, 1904 = Scythris noricella (Zeller, 1843); Scythris pacifica McDunnough, 1927 = Scythris immaculatella (Chambers, 1875); Coleophora albacostella Chambers, 1875 and Scythris hemidictyas Meyrick, 1928 = Neoscythris planipenella (Chambers, 1875).A cladistic definition of the family is presented for the first time. The monophyly of the Scythrididae is supported by the following synapomorphies: very narrow ductus bursae, broad ductus seminalis anastomosed with the oviduct and the corpus bursae, lack of signum, unique shape of the apophyses of the metathoracic furca, tarsomeres 1–4 with two subapical spurs, aedeagus ankylosed, and origin of forewing veins R4 and R5 on a common stalk with R4 extended to the costa and R5 to the termen. Relationships of the Scythrididae within the Gelechioidea are discussed. Based on the cladistic analysis of 52 structural characters, phylogenetic relationships of supraspecific taxa are inferred. Two cladograms, one for the genera and one for the species groups of Arotrura, are presented and used in deriving the classification.

BackgroundYponomeutoidea, one of the early-diverging lineages of ditrysian Lepidoptera, comprise about 1,800 species worldwide, including notable pests and insect-plant interaction models. Yponomeutoids were one of the earliest lepidopteran clades to evolve external feeding and to extensively colonize herbaceous angiosperms. Despite the group’s economic importance, and its value for tracing early lepidopteran evolution, the biodiversity and phylogeny of Yponomeutoidea have been relatively little studied.Methodology/Principal FindingsEight nuclear genes (8 kb) were initially sequenced for 86 putative yponomeutoid species, spanning all previously recognized suprageneric groups, and 53 outgroups representing 22 families and 12 superfamilies. Eleven to 19 additional genes, yielding a total of 14.8 to 18.9 kb, were then sampled for a subset of taxa, including 28 yponomeutoids and 43 outgroups. Maximum likelihood analyses were conducted on data sets differing in numbers of genes, matrix completeness, inclusion/weighting of synonymous substitutions, and inclusion/exclusion of “rogue” taxa. Monophyly for Yponomeutoidea was supported very strongly when the 18 “rogue” taxa were excluded, and moderately otherwise. Results from different analyses are highly congruent and relationships within Yponomeutoidea are well supported overall. There is strong support overall for monophyly of families previously recognized on morphological grounds, including Yponomeutidae, Ypsolophidae, Plutellidae, Glyphipterigidae, Argyresthiidae, Attevidae, Praydidae, Heliodinidae, and Bedelliidae. We also assign family rank to Scythropiinae (Scythropiidae stat. rev.), which in our trees are strongly grouped with Bedelliidae, in contrast to all previous proposals. We present a working hypothesis of among-family relationships, and an informal higher classification. Host plant family associations of yponomeutoid subfamilies and families are non-random, but show no trends suggesting parallel phylogenesis. Our analyses suggest that previous characterizations of yponomeutoids as predominantly Holarctic were based on insufficient sampling.Conclusions/SignificanceWe provide the first robust molecular phylogeny for Yponomeutoidea, together with a revised classification and new insights into their life history evolution and biogeography.

The known Lepidoptera (moths and butterflies) of the provinces and territories of Canada are summarised, and current knowledge is compared to the state of knowledge in 1979. A total of 5405 species are known to occur in Canada in 81 families, and a further 50 species have been reported but are unconfirmed. This represents an increase of 1348 species since 1979. The DNA barcodes available for Canadian Lepidoptera are also tabulated, based on a dataset of 148,314 specimens corresponding to 5842 distinct clusters. A further yet-undiscovered 1400 species of Lepidoptera are estimated to occur in Canada. The Gelechioidea are the most poorly known major lineage of Lepidoptera in Canada. Nunavut, Prince Edward Island, and British Columbia are thought to show the greatest deficit in our knowledge of Lepidoptera. The unglaciated portions of the Yukon (Beringia), and the Pacific Maritime, Montane Cordillera, and Western Interior Basin ecozones of British Columbia are also identified as hotbeds of undescribed biodiversity.