A switch to feeding on cycads generates parallel accelerated evolution of toxin tolerance in two clades of Eumaeus caterpillars (Lepidoptera: Lycaenidae).

We assembled a complete reference genome of Eumaeus atala, an aposematic cycad-eating hairstreak butterfly that suffered near extinction in the United States in the last century. Based on an analysis of genomic sequences of Eumaeus and 19 representative genera, the closest relatives of Eumaeus are Theorema and Mithras We report natural history information for Eumaeus, Theorema, and Mithras Using genomic sequences for each species of Eumaeus, Theorema, and Mithras (and three outgroups), we trace the evolution of cycad feeding, coloration, gregarious behavior, and other traits. The switch to feeding on cycads and to conspicuous coloration was accompanied by little genomic change. Soon after its origin, Eumaeus split into two fast evolving lineages, instead of forming a clump of close relatives in the phylogenetic tree. Significant overlap of the fast evolving proteins in both clades indicates parallel evolution. The functions of the fast evolving proteins suggest that the caterpillars developed tolerance to cycad toxins with a range of mechanisms including autophagy of damaged cells, removal of cell debris by macrophages, and more active cell proliferation.

Genomics-based higher classification of the species-rich Hairstreaks (Lepidoptera: Lycaenidae: Eumaeini).

We propose a higher classification of the lycaenid hairstreak tribe Eumaeini - one of the youngest and most species-rich butterfly tribes - based on autosome, Lepidopteran Z sex chromosome, and mitochondrial protein-coding genes. The subtribe Neolycaenina Korb is a synonym of Callophryidina Tutt, and subtribe Tmolusina Bálint is a synonym of Strephonotina K. Johnson, Austin, Le Crom, & Salazar. Proposed names are Rhammina Prieto & Busby, new subtribe; Timaetina Busby & Prieto, new subtribe; Atlidina Martins & Duarte, new subtribe; Evenina Faynel & Grishin, new subtribe; Jantheclina Robbins & Faynel, new subtribe; Paiwarriina Lamas & Robbins, new subtribe; Cupatheclina Lamas & Grishin, new subtribe; Parrhasiina Busby & Robbins, new subtribe; Ipideclina Martins & Grishin, new subtribe; and Trichonidina Duarte & Faynel, new subtribe. Phylogenetic results from the autosome and Z sex chromosome analyses are similar. Future analyses of datasets with hundreds of terminal taxa may be more practical time-wise by focussing on the smaller number of sex chromosome sequences (2.6% of nuclear protein-coding sequences). The phylogenetic classification and biological summaries for each subtribe suggest that a variety of factors affected Eumaeini diversification. About a dozen kinds of male secondary sexual organs with frequent evolutionary gains and losses occur in Atlidina, Evenina, and Jantheclina (141 species combined). Females have been shown to use these organs to discriminate between conspecific and non-conspecific males, facilitating sympatry among close relatives. Eumaeina, Rhammina, and Timaetina (140 species combined) are overwhelmingly montane with some evidence for a higher incidence of sympatric diversification. Seven Neotropical lineages in five subtribes invaded the temperate parts of the Nearctic Region with a diversification increase in the Callophryidina (262 species). North American Satyrium and Callophrys then invaded the Palearctic at least once each, with a major species-richness increase in Satyrium. The evolution of litter feeding detritivores within Calycopidina (172 species) resulted in an increase in diversification rate compared with its flower-feeding sister lineage. Atlidina, Strephonotina, Parrhasiina, and Strymonina (562 species combined) each contain a mixture of genera that specialize on one or two caterpillar food plant families and genera that are polyphagous. These would be appropriate subtribes to assess how the breadth of caterpillar food plants and the frequency of host shifts affected diversification.

Two new Thereus species from Peru, with notes on ecological niche partitioning (Lepidoptera: Lycaenidae: Theclinae).

Male hairstreaks (Theclinae) are often territorial, known for occupying their observation posts at a specific time during the day and at a certain height in the vegetation. The time and height for their perch vary from species to species but remain surprisingly stable year after year in any particular location. From these strategic observation posts males can check the passage of virgin females and chase away competing males. Consequently, these posts act as resources for the males and are not only vigorously defended from conspecific males but also form males of other species. In Peru, males of two closely related species belonging to the genus Thereus Hübner were observed in situ, defending their territories. One of them, Thereus tierralinda Faynel & Fåhraeus, sp. nov., is described here based on five male specimens from Cosñipata valley, Cusco, southern Peru. Further, it is compared to Thereus columbicola (Strand) and other close species in the Thereus eryssus species group. Observations over several years of T. columbicola and this new species show that they have different spatial and temporal characteristics for their perches, presumably to avoid competition with one another. Additionally, as a result of the partial revision of the group, a second new species, Thereus vicens Faynel & Fåhraeus, sp. nov., is also described based on one male and two females from Tingo María, Huánuco, central Peru. We also note that French Guiana is a new country record for Thereus eryssus (Herbst).

Integrative description of two new species and two new subspecies of Lamprospilus Geyer (Lepidoptera: Lycaenidae).

Lamprospilus stegmaier Prieto & Faynel sp. nov. is described from five specimens collected in sympatry with L. nicetus C. Felder & R. Felder on the northern central range of the Colombian Andes. Lamprospilus bicolor Faynel & Prieto sp. nov. is described from specimens collected in Colombia, Peru and Bolivia. Lamprospilus bicolor mirador Faynel & Prieto ssp. nov. is described from specimens collected in Peru and Bolivia. Lamprospilus decorata valluna Prieto & Faynel ssp. nov. is described from specimens collected in Western Colombia. Male and female phenotypes are associated, and we present morphological and molecular diagnostic characters for the new species. We also evaluate the congruence between a priori morphology-based species identifications and Molecular Operational Taxonomic Units (MOTUs) delimitations based on Barcode Index Numbers (BINs) in the genus. DNA barcodes are in perfect agreement with morphology in 64% of the species. Interspecific distances were found to range from 3.22% to 8.42% (average 5.48%), whilst their mean intraspecific variation ranges from 0.0% to 4.30% (average 1.78%). Based on DNA barcodes evidence of topotypic individuals, the study of original descriptions and illustrations of the holotypes, we consider here that Lamprospilus occidentalis Johnson & Salazar 2004 is a new junior subjective synonym of L. draudti Lathy, 1932.

Congruence between morphology-based species and Barcode Index Numbers (BINs) in Neotropical Eumaeini (Lycaenidae).

BACKGROUND: With about 1,000 species in the Neotropics, the Eumaeini (Theclinae) are one of the most diverse butterfly tribes. Correct morphology-based identifications are challenging in many genera due to relatively little interspecific differences in wing patterns. Geographic infraspecific variation is sometimes more substantial than variation between species. In this paper we present a large DNA barcode dataset of South American Lycaenidae. We analyze how well DNA barcode BINs match morphologically delimited species. METHODS: We compare morphology-based species identifications with the clustering of molecular operational taxonomic units (MOTUs) delimitated by the RESL algorithm in BOLD, which assigns Barcode Index Numbers (BINs). We examine intra- and interspecific divergences for genera represented by at least four morphospecies. We discuss the existence of local barcode gaps in a genus by genus analysis. We also note differences in the percentage of species with barcode gaps in groups of lowland and high mountain genera. RESULTS: We identified 2,213 specimens and obtained 1,839 sequences of 512 species in 90 genera. Overall, the mean intraspecific divergence value of CO1 sequences was 1.20%, while the mean interspecific divergence between nearest congeneric neighbors was 4.89%, demonstrating the presence of a barcode gap. However, the gap seemed to disappear from the entire set when comparing the maximum intraspecific distance (8.40%) with the minimum interspecific distance (0.40%). Clear barcode gaps are present in many genera but absent in others. From the set of specimens that yielded COI fragment lengths of at least 650 bp, 75% of the a priori morphology-based identifications were unambiguously assigned to a single Barcode Index Number (BIN). However, after a taxonomic a posteriori review, the percentage of matched identifications rose to 85%. BIN splitting was observed for 17% of the species and BIN sharing for 9%. We found that genera that contain primarily lowland species show higher percentages of local barcode gaps and congruence between BINs and morphology than genera that contain exclusively high montane species. The divergence values to the nearest neighbors were significantly lower in high Andean species while the intra-specific divergence values were significantly lower in the lowland species. These results raise questions regarding the causes of observed low inter and high intraspecific genetic variation. We discuss incomplete lineage sorting and hybridization as most likely causes of this phenomenon, as the montane species concerned are relatively young and hybridization is probable. The release of our data set represents an essential baseline for a reference library for biological assessment studies of butterflies in mega diverse countries using modern high-throughput technologies an highlights the necessity of taxonomic revisions for various genera combining both molecular and morphological data.

Cunoniaceae, nuevo registro de familia de plantas hospederas para los géneros altoandinos Penaincisalia y Micandra (Lycaenidae: Theclinae: Eumaeini) / Cunoniaceae, new family food plant record for the high altitude Andean genera Penaincisalia and Micandra (Lycaenidae: Theclinae: Eumaeini)

Resumen Orugas de dos especies altoandinas de Eumaeini (Lycaenidae: Theclinae) fueron halladas consumiendo hojas de Weinmannia crassifolia Ruiz & Pav., 1802 (Cunoniaceae). La colecta se realizó usando la técnica de colecta por golpeo. Esta familia de plantas es registrada por primera vez como planta alimenticia de los Eumaeini. Aunque no pudimos completar la crianza, las especies de Lycaenidae fueron identificadas mediante sus códigos de barra de ADN. Las orugas de Penaincisalia purpurea (K. Johnson, 1992) y Micandra dignota (Draudt, 1919) se describen e ilustran por primera vez. También se presentan datos sobre su ecología y relaciones con especies similares. Una tercera especie, registrada en estadio pupal en la misma planta de W. crassifolia fue un Nymphalidae, Perisama diotima (Hewitson, 1852).

Abstract Caterpillars of two Andean species of Eumaeini (Lycaenidae: Theclinae) were found eating leaves of Weinmannia crassifolia Ruiz & Pav., 1802 (Cunoniaceae). They were collected by "beating" the leaves. This plant family is reported for the first time as a food plant for Eumaeini. Although it was not possible to rear the caterpillars to the adult stage, we identified them using DNA barcodes. The caterpillars of Penaincisalia purpurea (K. Johnson, 1992) and Micandra dignota (Draudt, 1919) are described and illustrated for the first time. Data on their ecology and relationships with similar species are also presented. A third species collected on the same plant of W. crassifolia in the pupal stage was a Nymphalidae, Perisama diotima (Hewitson, 1852).

Crianza de Leptotes callanga (Lepidoptera : Lycaeni- dae : Polyommatinae) sobre Genista monspessulana (Fabaceae introducida de origen mediterráneo) en el Sur de Perú / Leptotes callanga (Lepidoptera : Lycaenidae : Polyommatinae) breeding on Genista monspessulana (Fabaceae introdu- ced of Mediterranean origin) in South Perú

Resumen La estación científica de Wayqecha se sitúa en el departamento de Cusco, en el sudeste de Perú, a una altitud de 2900 m (13°10.506'S - 71°35.210'W). En esta estación, encontramos una oruga de Leptotes callanga (Dyar, 1913) (Lepidoptera : Lycaenidae : Polyommatinae), alimentándose de flores de Genista monspessulana (L.) L.A.S. Johnson (Fabaceae), una planta no nativa, de origen mediterráneo. La crianza permitió obtener una hembra adulta de L. callanga. La oruga en el último estadio y la crisálida se describen e ilustran. Se comenta la adaptación de esta especie a una nueva planta.

Abstract The Wayqecha scientific station is located in the department of Cusco, in southeas- tern Peru, at an altitude of 2900 m (13°10.506'S - 71°35.210'W). In this station, we found a caterpillar of Leptotes callanga (Dyar, 1913) (Lepidoptera : Lycaenidae : Polyommatinae), consuming flowers of Genista monspessulana (L.) L.A.S. Johnson (Fabaceae), a non- native plant of Mediterranean origin. The breeding allowed to obtain a female L. callanga. Last instar larvae and pupa are described and illustrated. Leptotes callanga adaptation to this new plant is commented.

Crianza de Michaelus phoenissa (Lepidoptera: Lycaenidae) sobre Senna alata (Fabaceae) en Perú / Breeding of Michaelus phoenissa (Lepidoptera : Lycaenidae) on Senna alata (Fabaceae) in Peru

En la estación científica de Villa Carmen, situada cerca de Pillcopata, Cusco, Perú (S 12°53.687' - W 71°24.232', 533 m), encontramos orugas de Michaelus phoenissa (Hewitson, 1867) (Lycaenidae), consumiendo flores de Senna alata (L.) Roxb. (Fabaceae). La mayoría de las orugas se encontraban en el último estadio (entre 1 y 1.5 cm de longitud), no visibles a primera vista, ya que se alojaban en las flores no abiertas. La crianza permitió obtener cuatro adultos, dos hembras y dos machos y ningún parasitoide. Se compara las plantas hospederas de M. phoenissa con otras especies hospederas del genero de Michaelus Nicolay, 1979.

In Villa Carmen, a scientific station near Pillcopata, Cusco, Peru (S 12°53.687' - W 71°24.232', 533 m), we found caterpillars of Michaelus phoenissa (Hewitson, 1867) (Lycaenidae) consuming flowers of Senna alata (L.) Roxb. (Fabaceae). The caterpillars were mostly last instars (between 1 and 1.5 cm in length). They were not visible at first sight because they lodged in unopened flowers. We reared two female and two male adults. None of the immature stages was parasitized. We compare the food plants of M. phoenissa with those of others species of Michaelus Nicolay, 1979.

Review of the species level taxonomy of the neotropical butterfly genus Oenomaus (Lycaenidae, Theclinae, Eumaeini).

Seven new species of the Neotropical hairstreak genus Oenomaus are described: Oenomaus mancha Busby & Faynel, sp. n. (type locality Ecuador); Oenomaus gwenish Robbins & Faynel, sp. n. (type locality Panama); Oenomaus lea Faynel & Robbins, sp. n. (type locality Ecuador); Oenomaus myrteana Busby, Robbins & Faynel, sp. n. (type locality Ecuador); Oenomaus mentirosa Faynel & Robbins, sp. n. (type locality Peru); Oenomaus andi Busby & Faynel, sp. n. (type locality Ecuador) and Oenomaus moseri Robbins & Faynel, sp. n. (type locality Brazil, Santa Catarina). For each new Oenomaus species, we present diagnostic characters and notes on its habitat and biology. We illustrate adults, genitalia, and distribution. New distributional and biological data are presented for 21 previously described Oenomaus species. Oenomaus melleus guyanensis Faynel, 2008 is treated as a new synonym of Oenomaus melleus melleus (Druce, 1907). Females are described and associated with males for ten species using a variety of factors, including mitochondrial COI DNA "barcode" sequences. We summarize the reasons why the number of recognized Oenomaus species has grown in the past decade from one species to 28 species. Finally, we overview the habitats that Oenomaus species occupy and note that the agricultural pest on Annonaceae, Oenomaus ortygnus, is the only Oenomaus species that regularly occurs in greatly disturbed habitats.