@article {22968, title = {A high-quality, long-read genome assembly of the whitelined sphinx moth (Lepidoptera: Sphingidae: Hyles lineata) shows highly conserved melanin synthesis pathway genes}, journal = {G3 Genes|Genomes|Genetics}, volume = {13}, year = {2023}, month = {04/2023}, pages = {jkad090}, chapter = {jkad090}, abstract = {

\"The sphinx moth genus Hyles comprises 29 described species inhabiting all continents except Antarctica. The genus diverged relatively recently (40\–25 MYA), arising in the Americas and rapidly establishing a cosmopolitan distribution. The whitelined sphinx moth, Hyles lineata, represents the oldest extant lineage of this group and is one of the most widespread and abundant sphinx moths in North America. Hyles lineata exhibits the large body size and adept flight control characteristic of the sphinx moth family (Sphingidae), but it is unique in displaying extreme larval color variation and broad host plant use. These traits, in combination with its broad distribution and high relative abundance within its range, have made H. lineata a model organism for studying phenotypic plasticity, plant\–herbivore interactions, physiological ecology, and flight control. Despite being one of the most well-studied sphinx moths, little data exist on genetic variation or regulation of gene expression. Here, we report a high-quality genome showing high contiguity (N50 of 14.2 Mb) and completeness (98.2\% of Lepidoptera BUSCO genes), an important first characterization to facilitate such studies. We also annotate the core melanin synthesis pathway genes and confirm that they have high sequence conservation with other moths and are most similar to those of another, well-characterized sphinx moth, the tobacco hornworm (Manduca sexta).\"

}, keywords = {DNA, GENETICS, GENOME ASSEMBLY, HYLES, MANDUCA, MELANIN SYNTHESIS, RNA, SPHINGIDAE}, doi = {10.1093/g3journal/jkad090}, url = {https://doi.org/10.1093/g3journal/jkad090}, author = {Godfrey, R.K. and Britton, S.E. and Mishra, S. and Goldberg, J.K. and Kawahara, A.Y.} } @article {22562, title = {Hidden phylogenomic signal helps elucidate arsenurine silkmoth phylogeny and the evolution of body size and wing shape trade-offs}, journal = {Systematic Biology}, volume = {71}, year = {2022}, month = {06/2022}, pages = {859{\textendash}874}, chapter = {859}, abstract = {

\"One of the key objectives in biological research is understanding how evolutionary processes have produced Earth\’s diversity. A critical step toward revealing these processes is an investigation of evolutionary tradeoffs\—that is, the opposing pressures of multiple selective forces. For millennia, nocturnal moths have had to balance successful flight, as they search for mates or host plants, with evading bat predators. However, the potential for evolutionary trade-offs between wing shape and body size are poorly understood. In this study, we used phylogenomics and geometric morphometrics to examine the evolution of wing shape in the wild silkmoth subfamily Arsenurinae (Saturniidae) and evaluate potential evolutionary relationships between body size and wing shape. The phylogeny was inferred based on 782 loci from target capture data of 42 arsenurine species representing all 10 recognized genera. After detecting in our data one of the most vexing problems in phylogenetic inference\—a region of a tree that possesses short branches and no \“support\” for relationships (i.e., a polytomy), we looked for hidden phylogenomic signal (i.e., inspecting differing phylogenetic inferences, alternative support values, quartets, and phylogenetic networks) to better illuminate the most probable generic relationships within the subfamily. We found there are putative evolutionary trade-offs between wing shape, body size, and the interaction of fore- and hindwing (HW) shape. Namely, body size tends to decrease with increasing HW length but increases as forewing (FW) shape becomes more complex. Additionally, the type of HW (i.e., tail or no tail) a lineage possesses has a significant effect on the complexity of FW shape. We outline possible selective forces driving the complex HW shapes that make Arsenurinae, and silkmoths as a whole, so charismatic.\"

}, keywords = {ANCHORED HYBRID ENRICHMENT, ARSENURINAE, GEOMETRIC MORPHOMETRICS, PHYLOGENOMICS, SATURNIIDAE, TRADE-OFFS, TRAITS, WING SHAPE}, doi = {10.1093/sysbio/syab090}, author = {Hamilton, C.A. and Winiger, N. and Rubin, J.J. and Breinholt, J. and Rougerie, R. and Kitching, I.J. and Barber, J.R. and Kawahara, A.Y.} } @article {22476, title = {A diversification relay race from Caribbean-Mesoamerica to the Andes: historical biogeography of Xylophanes hawkmoths}, journal = {Proceedings of the Royal Society B}, volume = {289}, year = {2022}, month = {02/2022}, pages = {20212435}, chapter = {20212435}, abstract = {

\"The regions of the Andes and Caribbean-Mesoamerica are both hypothesized to be the cradle for many Neotropical lineages, but few studies have fully investigated the dynamics and interactions between Neotropical bioregions. The NewWorld hawkmoth genus Xylophanes is the most taxonomically diverse genus in the Sphingidae, with the highest endemism and richness in the Andes and Caribbean-Mesoamerica. We integrated phylogenomic and DNA barcode data and generated the first time-calibrated tree for this genus, covering 93.8\% of the species diversity. We used event-based likelihood ancestral area estimation and biogeographic stochastic mapping to examine the speciation and dispersal dynamics of Xylophanes across bioregions. We also used trait-dependent diversification models to compare speciation and extinction rates of lineages associated with different bioregions. Our results indicate that Xylophanes originated in Caribbean-Mesoamerica in the Late Miocene, and immediately diverged into five major clades. The current species diversity and distribution of Xylophanes can be explained by two consecutive phases. In the first phase, the highest Xylophanes speciation and emigration rates occurred in the Caribbean-Mesoamerica, and the highest immigration rates occurred in the Andes, whereas in the second phase the highest immigration rates were found in Amazonia, and the Andes had the highest speciation and emigration rates.\"

}, keywords = {ANCESTRAL AREAS ANALYSIS, ANDES, BARCODING, BioGeoBEARS, BIOGEOGRAPHY, CARIBBEAN, COI, DIVERSIFICATION, DNA, EMIGRATION RATES, EVOLUTION, MAXIMUM LIKELIHOOD, MESOAMERICA, MODELS, PHYLOGENOMICS, PHYLOGENY, SPECIATION, SPHINGIDAE, STOCHASTIC MAPPING, XYLOPHANES}, doi = {10.1098/rspb.2021.2435}, url = {https://doi.org/10.1098/rspb.2021.2435}, author = {Li, X. and Hamilton, C.A. and St Laurent, R.A. and Ballesteros-Mejia, L. and Markee, A. and Haxaire, J. and Rougerie, R. and Kitching, I.J. and Kawahara, A.Y.} } @article {22414, title = {The evolution of two distinct strategies of moth flight}, journal = {Journal of the Royal Society Interface}, volume = {18}, year = {2021}, month = {11/2021}, pages = {20210632}, chapter = {202106v32}, abstract = {

\"Across insects, wing shape and size have undergone dramatic divergence even in closely related sister groups. However, we do not know how morphology changes in tandem with kinematics to support body weight within available power and how the specific force production patterns are linked to differences in behaviour. Hawkmoths and wild silkmoths are diverse sister families with divergent wing morphology. Using three-dimensional kinematics and quasisteady aerodynamic modelling,we compare the aerodynamics and the contributions of wing shape, size and kinematics in 10 moth species. We find that wing movement also diverges between the clades and underlies two distinct strategies for flight.Hawkmoths use wing kinematics, especially high frequencies, to enhance force and wing morphologies that reduce power. Silkmoths use wing morphology to enhance force, and slow, high-amplitude wingstrokes to reduce power. Both strategies converge on similar aerodynamic power and can support similar body weight ranges. However, inter-clade within-wingstroke force profiles are quite different and linked to the hovering flight of hawkmoths and the bobbing flight of silkmoths. These two moth groups fly more like other, distantly related insects than they do each other, demonstrating the diversity of flapping flight evolution and a rich bioinspired design space for robotic flappers.\"

}, keywords = {AERODYNAMICS, EVOLUTION, FLIGHT CONTROL, KINEMATICS, MORPHOLOGY, MORPHOMETRICS, SATURNIIDAE, SPHINGIDAE, WING LOADING, WING SHAPE}, doi = {10.1098/rsif.2021.0632}, url = {https://doi.org/10.1098/rsif.2021.0632}, author = {Aiello, B.R. and Sikandar, U.B. and Minoguchi, H. and Bhinderwala, B. and Hamilton, C.A. and Kawahara, A.Y. and Sponberg, S.} } @unpublished {22378, title = {Anti-bat ultrasound production in moths is globally and phylogenetically widespread}, year = {2021}, month = {09/2021}, pages = {1-31}, type = {bioRxiv preprint}, abstract = {

\"Warning signals are well known in the visual system, but rare in other modalities. Some moths produce ultrasonic sounds to warn bats of noxious taste or to mimic unpalatable models. Here we report results from a long-term study across the globe, assaying moth response to playback of bat echolocation. We tested 252 genera, spanning most families of large-bodied moths, and outline anti-bat ultrasound production in 52 genera, with eight new subfamily origins described. Based on acoustic analysis of ultrasonic emissions and palatability experiments with bats, it seems that acoustic warning and mimicry are the raison d\&$\#$39;etre for sound production in most moths. However, some moths use high-density ultrasound capable of jamming bat sonar. In fact, we find preliminary evidence of independent origins of sonar jamming in at least six subfamilies. Palatability data indicates that jamming and warning are not mutually exclusive strategies. To explore the possible organization of anti-bat warning sounds into acoustic mimicry rings, we intensively studied a community of moths in Ecuador and found five distinct acoustic clusters using machine learning algorithms. While these data represent an early understanding of acoustic aposematism and mimicry across this megadiverse insect order, it is likely that ultrasonically-signaling moths comprise one of the largest mimicry complexes on earth.\"

}, keywords = {ACOUSTICS, APOSEMATISM, BATS, DEFENCE, DIVERSITY, EREBIDAE, EVOLUTION, HEARING, JAMMING, LEPIDOPTERA, MIMICRY, PREDATOR AVOIDANCE, SPHINGIDAE, STRIDULATION, TYMBAL ORGANS, ULTRASONICS}, doi = {10.1101/2021.09.20.460855}, url = {https://doi.org/10.1101/2021.09.20.460855}, author = {Barber, J.R. and Plotkin, D. and Rubin, J.J. and Homziak, N.T. and Leavell, B.C. and Houlihan, P. and Miner, K.A. and Breinholt, J.W. and Quirk-Royal, B. and Padr{\'o}n, P.S. and Nunez, M. and Kawahara, A.Y.} } @article {22377, title = {Adaptive shifts underlie the divergence in wing morphology in bombycoid moths}, journal = {Proceedings of the Royal Society B}, volume = {288}, year = {2021}, month = {08/2021}, pages = {20210677}, chapter = {20210677}, abstract = {

\"The evolution of flapping flight is linked to the prolific success of insects. Across Insecta, wing morphology diversified, strongly impacting aerodynamic performance. In the presence of ecological opportunity, discrete adaptive shifts and early bursts are two processes hypothesized to give rise to exceptional morphological diversification. Here, we use the sister-families Sphingidae and Saturniidae to answer how the evolution of aerodynamically important traits is linked to clade divergence and through what process(es) these traits evolve. Many agile Sphingidae evolved hover-feeding behaviors, while adult Saturniidae lack functional mouth parts and rely on a fixed energy budget as adults. We find that Sphingidae underwent an adaptive shift in wing morphology coincident with life history and behavior divergence, evolving small high aspect-ratio wings advantageous for power reduction that can be moved at high frequencies, beneficial for flight control. In contrast, Saturniidae, which do not feed as adults, evolved large wings and morphology which surprisingly does not reduce aerodynamic power, but could contribute to their erratic flight behavior, aiding in predator avoidance. We suggest that after the evolution of flapping flight, diversification of wing morphology can be potentiated by adaptative shifts, shaping the diversity of wing morphology across insects.\"

}, keywords = {ADAPTATION, AERODYNAMICS, ASPECT RATIO, BOMBYCOIDEA, EVOLUTION, FLIGHT CONTROL, SATURNIIDAE, SPHINGIDAE}, doi = {10.1098/rspb.2021.0677}, url = {https://doi.org/10.1098/rspb.2021.0677}, author = {Aiello, B.R. and Tan, M. and Bin Sikandar, U. and Alvey, A.J. and Bhinderwala B. and Kimball, K.C. and Barber, J.R. and Hamilton, C.A. and Kawahara, A.Y. and Sponberg, S.} } @article {22244, title = {Population differentiation and structural variation in the Manduca sexta genome across the United States}, journal = {G3 Genes|Genomes|Genetics}, volume = {12}, year = {2022}, month = {02/2022}, pages = {jkac047}, chapter = {jkac047}, abstract = {

\"Many species that are extensively studied in the laboratory are less well characterized in their natural habitat, and laboratory strains represent only a small fraction of the variation in a species\’ genome. Here we investigate genomic variation in three natural populations of an agricultural pest and a model insect for many scientific disciplines, the tobacco hornworm (Manduca sexta). We show that hornworms from Arizona, Kansas, and North Carolina are genetically distinct, with Arizona being particularly differentiated from the other two populations. Specifically, two segregating inversions and a potential pseudogene are found only in the Arizona population. One inversion on the Z chromosome may enhance adaptive evolution of the sex chromosome, while the significance of the other, autosomal inversion remains unclear. The pseudogene may be involved in the exploitation of a novel hostplant in Arizona, but functional genetic assays will be required to confirm this hypothesis. Nevertheless, our results reveal undiscovered natural variation and provide useful genomic data for a model insect species.\"

}, keywords = {BIOGEOGRAPHY, CHROMOSOME INVERSION, CHROMOSOMES, DNA, MANDUCA, PHYLOGENOMICS, POPULATION BIOLOGY, POPULATION GENETICS, PSEUDOGENE, SEX CHROMOSOMES, SPHINGIDAE, USA}, doi = {10.1093/g3journal/jkac047}, url = {https://doi.org/10.1093/g3journal/jkac047}, author = {Mongue, A.J. and Kawahara, A.Y.} } @article {22093, title = {Phylogeny of the hawkmoth tribe Ambulycini (Lepidoptera: Sphingidae): mitogenomes from museum specimens resolve major relationships.}, journal = {Insect Systematics and Diversity}, volume = {3}, year = {2019}, month = {12/2019}, pages = {12, 1-8}, chapter = {12}, abstract = {

\"Ambulycini are a cosmopolitan tribe of the moth family Sphingidae, comprised of 10 genera, 3 of which are found in tropical Asia, 4 in the Neotropics, 1 in Africa, 1 in the Middle East, and 1 restricted to the islands of New Caledonia. Recent phylogenetic analyses of the tribe have yielded conflicting results, and some have suggested a close relationship of the monobasic New Caledonian genus Compsulyx Holloway, 1979 to the Neotropical ones, despite being found on opposite sides of the Pacific Ocean. Here, we investigate relationships within the tribe using full mitochondrial genomes, mainly derived from dry-pinned museum collections material. Mitogenomic data were obtained for 19 species representing nine of the 10 Ambulycini genera. Phylogenetic trees are in agreement with a tropical Asian origin for the tribe. Furthermore, results indicate that the Neotropical genus Adhemarius Oiticica Filho, 1939 is paraphyletic and support the notion that Orecta Rothschild \& Jordan 1903 and Trogolegnum Rothschild \& Jordan, 1903 may need to be synonymized. Finally, in our analysis the Neotropical genera do not collectively form a monophyletic group, due to a clade comprising the New Caledonian genus Compsulyx and the African genus Batocnema Rothschild \& Jordan, 1903 being placed as sister to the Neotropical genus Protambulyx Rothschild \& Jordan, 1903. This finding implies a complex biogeographic history and suggests the evolution of the tribe involved at least two long-distance dispersal events.\"

}, keywords = {ADHEMARIUS, AMBULYCINI, BATOCNEMA, BAYESIAN INFERENCE, BIOGEOGRAPHY, COMPSULYX, MAXIMUM LIKELIHOOD, MITOGENOME, NEXTGEN SEQUENCING, PHYLOGENOMICS, PHYLOGENY, PROTAMBULYX, SPHINGIDAE, TROGOLEGNUM}, doi = {10.1093/isd/ixz025}, author = {Timmermans, M.J.T.N. and Daghmoumi, S.M. and Glass, D. and Hamilton, C.A. and Kawahara, A.Y. and Kitching, I.J.} } @article {22026, title = {Phylogenomics resolves major relationships and reveals significant diversification rate shifts in the evolution of silk moths and relatives.}, journal = {BMC Evolutionary Biology}, volume = {19}, year = {2019}, month = {09/2019}, pages = {182 [1-13]}, chapter = {182}, abstract = {

\"Background: Silkmoths and their relatives constitute the ecologically and taxonomically diverse superfamily Bombycoidea, which includes some of the most charismatic species of Lepidoptera. Despite displaying spectacular forms and diverse ecological traits, relatively little attention has been given to understanding their evolution and drivers of their diversity. To begin to address this problem, we created a new Bombycoidea-specific Anchored Hybrid Enrichment (AHE) probe set and sampled up to 571 loci for 117 taxa across all major lineages of the Bombycoidea, with a newly developed DNA extraction protocol that allows Lepidoptera specimens to be readily sequenced from pinned natural history collections.
Results: The well-supported tree was overall consistent with prior morphological and molecular studies, although some taxa were misplaced. The bombycid Arotros Schaus was formally transferred to Apatelodidae. We identified important evolutionary patterns (e.g., morphology, biogeography, and differences in speciation and extinction), and our analysis of diversification rates highlights the stark increases that exist within the Sphingidae (hawkmoths) and Saturniidae (wild silkmoths).
Conclusions: Our study establishes a backbone for future evolutionary, comparative, and taxonomic studies of Bombycoidea. We postulate that the rate shifts identified are due to the well-documented bat-moth \“arms race\”. Our research highlights the flexibility of AHE to generate genomic data from a wide range of museum specimens, both age and preservation method, and will allow researchers to tap into the wealth of biological data residing in natural history collections around the globe.\"

}, keywords = {ANCHORED HYBRID ENRICHMENT, APATELODIDAE, AROTROS, BOMBYCIDAE, BOMBYCOIDEA, DNA, PHYLOGENOMICS, PHYLOGENY, TARGETED SEQUENCE CAPTURE}, doi = {10.1186/s12862-019-1505-1}, url = {https://doi.org/10.1186/s12862-019-1505-1}, author = {Hamilton, C.A. and St Laurent, R.A. and Dexter, K. and Kitching, I.J. and Breinholt, J.W. and Zwick, A. and Timmermans, M.J.T.N. and Barber, J.R. and Kawahara, A.Y.} } @article {21971, title = {Two New Species of Psychocampa and a Possible Case of Visual Mimicry in the Sack-Bearer Moths (Lepidoptera: Mimallonoidea)}, journal = {Insect Systematics and Diversity}, volume = {3}, year = {2019}, month = {04/2019}, pages = {3 [9 pp.]}, chapter = {3}, abstract = {

\"Within the Lepidoptera, diurnal adult behavior is more often associated with butterflies than moths. However, many moth species are also known to have adults that fly during the day, and the lack of diel activity data for moths can be attributed to limited natural history observations. Moths belonging to Mimallonidae Burmeister are one such example of a family of Lepidoptera for which very little is known about adult activity times, with literature reports of only two of the nearly 300 described species known to exhibit diurnal behavior. In the present study, two new species of black and white Psychocampa Grote \& Robinson are described: P. aello St Laurent, sp. n. from Colombia and P. celaeno St Laurent, sp. n. from Panama. We hypothesize that these two species are diurnal due to their rarity in collections and apparent similarity in appearance with diurnal sphingid hawkmoths of the genus Aellopos H\übner (Lepidoptera: Sphingidae) and certain Hymenoptera. Psychocampa funebris (Schaus) is the species most similar to the two new species described here, it is known from Argentina, southeastern Brazil, and Uruguay whereas the new species are from Panama and western Colombia. Psychocampa funebris is known to be diurnal, and therefore, it is likely that the two new, apparently related species described herein, are also diurnal. Considering the hypothesized diurnal behavior of the new species, and their unique coloring and patterning, we believe this to be the first documentation of potential adult mimicry in Mimallonidae.\"

}, keywords = {AELLOPOS, COLOMBIA, MIMALLONIDAE, MIMICRY, PANAMA, PSYCHOCAMPA, SPHINGIDAE}, url = {https://doi.org/10.1093/isd/ixz003}, author = {St. Laurent, R. and Carvalho, A.P.S. and Kawahara, A.Y.} } @article {21830, title = {A global checklist of the Bombycoidea (Insecta: Lepidoptera)}, journal = {Biodiversity Data Journal}, volume = {6}, year = {2018}, month = {02/2018}, pages = {e22236}, chapter = {e22236}, abstract = {

\"Background
Bombycoidea is an ecologically diverse and speciose superfamily of Lepidoptera. The superfamily includes many model organisms, but the taxonomy and classification of the superfamily has remained largely in disarray. Here we present a global checklist of Bombycoidea. Following Zwick (2008) and Zwick et al. (2011), ten families are recognized: Anthelidae, Apatelodidae, Bombycidae, Brahmaeidae, Carthaeidae, Endromidae, Eupterotidae, Phiditiidae, Saturniidae and Sphingidae. The former families Lemoniidae and Mirinidae are included within Brahmaeidae and Endromidae respectively. The former bombycid subfamilies Oberthueriinae and Prismostictinae are also treated as synonyms of Endromidae, and the former bombycine subfamilies Apatelodinae and Phitditiinae are treated as families.


New information
This checklist represents the first effort to synthesize the current taxonomic treatment of the entire superfamily. It includes 12,159 names and references to their authors, and it accounts for the recent burst in species and subspecies descriptions within family Saturniidae (ca. 1,500 within the past 10 years) and to a lesser extent in Sphingidae (ca. 250 species over the same period). The changes to the higher classification of Saturniidae proposed by N\ässig et al. (2015) are rejected as premature and unnecessary. The new tribes, subtribes and genera described by Cooper (2002) are here treated as junior synonyms. We also present a new higher classification of Sphingidae, based on Kawahara et al. (2009), Barber and Kawahara (2013) and a more recent phylogenomic study by Breinholt et al. (2017), as well as a reviewed genus and species level classification, as documented by Kitching (2018).\"

}, keywords = {ANTHELIDAE, APATELODIDAE, BOMBYCIDAE, BRAHMAEIDAE, CARTHAEIDAE, CATALOGUE, CHECKLIST, ENDROMIDAE, EUPTEROTIDAE, HIGHER CLASSIFICATION, ORECTA, PHIDITIIDAE, SATURNIIDAE, SPHINGIDAE}, doi = {10.3897/BDJ.6.e22236}, url = {https://doi.org/10.3897/BDJ.6.e22236}, author = {Kitching, I.J. and Rougerie, R. and Zwick, A. and Hamilton, C.A. and St._Laurent, R. and Naumann, S. and Ballesteros-Mejia, L. and Kawahara, A.Y.} } @article {21819, title = {Functional characterization of the Hyles euphorbiae hawkmoth transcriptome reveals strong expression of phorbol ester detoxification and seasonal cold hardiness genes}, journal = {Frontiers in Zoology}, volume = {15}, year = {2018}, month = {05/2018}, pages = {20}, chapter = {20}, abstract = {

\"Background: The European spurge hawkmoth, Hyles euphorbiae (Lepidoptera, Sphingidae), has been intensively studied as a model organism for insect chemical ecology, cold hardiness and evolution of species delineation. To understand species isolation mechanisms at a molecular level, this study aims at determining genetic factors underlying two adaptive ecological trait candidates, phorbol ester (TPA) detoxification and seasonal cold acclimation.
Method: A draft transcriptome of H. euphorbiae was generated using Illumina sequencing, providing the first genomic resource for the hawkmoth subfamily Macroglossinae. RNA expression levels in tissues of experimental TPA feeding larvae and cooled pupae was compared to levels in control larvae and pupae using 26 bp RNA sequence tag libraries (DeepSuperSAGE). Differential gene expression was assessed by homology searches of the tags in the transcriptome.
Results: In total, 389 and 605 differentially expressed transcripts for detoxification and cold hardiness, respectively, could be identified and annotated with proteins. The majority (22 of 28) of differentially expressed detox transcripts of the four \&$\#$39;drug metabolism\&$\#$39; enzyme groups (cytochrome P450 (CYP), carboxylesterases (CES), glutathione S-transferases (GST) and lipases) are up-regulated. Triacylglycerol lipase was significantly over proportionally annotated among up-regulated detox transcripts. We record several up-regulated lipases, GSTe2, two CESs, CYP9A21, CYP6BD6 and CYP9A17 as candidate genes for further H. euphorbiae TPA detoxification analyses. Differential gene expression of the cold acclimation treatment is marked by metabolic depression with enriched Gene Ontology terms among down-regulated transcripts almost exclusively comprising metabolism, aerobic respiration and dissimilative functions. Down-regulated transcripts include energy expensive respiratory proteins like NADH dehydrogenase, cytochrome oxidase and ATP synthase. Gene expression patterns show shifts in carbohydrate metabolism towards cryoprotectant production. The Glycolysis enzymes, G1Pase, A1e, Gpi and an Akr isoform are up-regulated. Glycerol, an osmolyte which lowers the body liquid supercooling point, appears to be the predominant polyol cryoprotectant in H. euphorbiae diapause pupae. Several protein candidates involved in glucose, glycerol, myo-inositol and potentially sorbitol and trehalose synthesis were identified.
Conclusions: A majority of differently expressed transcripts unique for either detoxification or cold hardiness indicates highly specialized functional adaptation which may have evolved from general cell metabolism and stress response.The transcriptome and extracted candidate biomarkers provide a basis for further gene expression studies of physiological processes and adaptive traits in H. euphorbiae.\"

}, keywords = {12-TETRADECANOYL-PHORBOL-13-ACETATE (TPA), DETOXIFICATION, DIAPAUSE, DITERPENE ESTER, ECOLOGY, FREEZING TOLERANCE, HYLES, PHORBOL ESTER, RNA, SPHINGIDAE, TRAITS, TRANSCRIPTOMICS}, doi = {10.1186/s12983-018-0252-2}, url = {https://doi.org/10.1186/s12983-018-0252-2}, author = {Barth, M.B. and Buchwalder, K. and Kawahara, A.Y. and Zhou, X. and Liu, S. and Krezdorn, N. and Rotter, B. and Horres, R. and Hundsdoerfer, A.K.} } @article {21525, title = {Resolving relationships among the megadiverse butterflies and moths with a novel pipeline for anchored phylogenomics}, journal = {Systematic Biology}, year = {2017}, type = {Early view}, abstract = {

\"The advent of next-generation sequencing technology has allowed for the collection of large portions of the genome for phylogenetic analysis. Hybrid enrichment and transcriptomics are two techniques that leverage next-generation sequencing and have shown much promise. However, methods for processing hybrid enrichment data are still limited. We developed a pipeline for anchored hybrid enrichment (AHE) read assembly, orthology determination, contamination screening, and data processing for sequences flanking the target \“probe\” region. We apply this approach to study the phylogeny of butterflies and moths (Lepidoptera), a megadiverse group of more than 157,000 described species with poorly understood deep-level phylogenetic relationships. We introduce a new, 855 locus AHE kit for Lepidoptera phylogenetics and compare resulting trees to those from transcriptomes. The enrichment kit was designed from existing genomes, transcriptomes, and expressed sequence tags and was used to capture sequence data from 54 species from 23 lepidopteran families. Phylogenies estimated from AHE data were largely congruent with trees generated from transcriptomes, with strong support for relationships at all but the deepest taxonomic levels.We combineAHEand transcriptomic data to generate a new Lepidoptera phylogeny, representing 76 exemplar species in 42 families. The tree provides robust support for many relationships, including those among the seven butterfly families. The addition of AHE data to an existing transcriptomic dataset lowers node support along the Lepidoptera backbone, but firmly places taxa with AHE data on the phylogeny. Combining taxa sequenced for AHE with existing transcriptomes and genomes resulted in a tree with strong support for (Calliduloidea + Gelechioidea + Thyridoidea) + (Papilionoidea + Pyraloidea + Macroheterocera). To examine the efficacy of AHE at a shallowtaxonomic level, phylogenetic analyseswere also conducted on a sister group representing a more recent divergence, the Saturniidae and Sphingidae. These analyses utilized sequences from the probe region and data flanking it, nearly doubled the size of the dataset; resulting trees supported new phylogenetics relationships, especially within the Saturniidae and Sphingidae (e.g., Hemarina derived in the latter). We hope that our data processing pipeline, hybrid enrichment gene set, and approach of combining AHE data with transcriptomes will be useful for the broader systematics community.\"

}, keywords = {ANCHORED HYBRID ENRICHMENT, HEMARINI, HIGHER CLASSIFICATION, LEPIDOPTERA, METHODOLOGY, PHYLOGENOMICS, PHYLOGENY, PIPELINE, SATURNIIDAE, SPHINGIDAE}, doi = {10.1093/sysbio/syx048}, author = {Breinholt, J.W. and Earl, C. and Lemmon, A.R. and Moriarty_Lemmon, E. and Xiao, L. and Kawahara, A.Y.} } @article {21513, title = {A large-scale, higher-Level, molecular phylogenetic study of the insect order Lepidoptera (moths and butterflies)}, journal = {PLoS ONE}, volume = {8}, year = {2013}, month = {03/2013}, pages = {e58568}, chapter = {e58568}, abstract = {

\"Background: Higher-level relationships within the Lepidoptera, and particularly within the species-rich subclade Ditrysia, are
generally not well understood, although recent studies have yielded progress. We present the most comprehensive
molecular analysis of lepidopteran phylogeny to date, focusing on relationships among superfamilies.

Methodology / Principal Findings: 483 taxa spanning 115 of 124 families were sampled for 19 protein-coding nuclear
genes, from which maximum likelihood tree estimates and bootstrap percentages were obtained using GARLI. Assessment
of heuristic search effectiveness showed that better trees and higher bootstrap percentages probably remain to be
discovered even after 1000 or more search replicates, but further search proved impractical even with grid computing.
Other analyses explored the effects of sampling nonsynonymous change only versus partitioned and unpartitioned total
nucleotide change; deletion of rogue taxa; and compositional heterogeneity. Relationships among the non-ditrysian
lineages previously inferred from morphology were largely confirmed, plus some new ones, with strong support. Robust
support was also found for divergences among non-apoditrysian lineages of Ditrysia, but only rarely so within Apoditrysia.
Paraphyly for Tineoidea is strongly supported by analysis of nonsynonymous-only signal; conflicting, strong support for
tineoid monophyly when synonymous signal was added back is shown to result from compositional heterogeneity.

Conclusions / Significance: Support for among-superfamily relationships outside the Apoditrysia is now generally strong.
Comparable support is mostly lacking within Apoditrysia, but dramatically increased bootstrap percentages for some nodes
after rogue taxon removal, and concordance with other evidence, strongly suggest that our picture of apoditrysian
phylogeny is approximately correct. This study highlights the challenge of finding optimal topologies when analyzing
hundreds of taxa. It also shows that some nodes get strong support only when analysis is restricted to nonsynonymous
change, while total change is necessary for strong support of others. Thus, multiple types of analyses will be necessary to
fully resolve lepidopteran phylogeny.\"

}, keywords = {109FIN, 265FIN, 268FIN, 3007FIN, ACC, APODITRYSIA, BARCODING, BOOTSTRAP, CAD, COI, DDC, DNA, ENOLASE, GARLI, HIGHER CLASSIFICATION, LEPIDOPTERA, MACROHETEROCERA, MAXIMUM LIKELIHOOD, MTDNA, PHYLOGENY, ROGUE TAXA, TINEOIDEA}, doi = {10.1371/journal.pone.0058568}, author = {Regier, J.C. and Mitter, C. and Zwick, A. and Bazinet, A.L. and Cumming, M.P. and Kawahara, A.Y. and Sohn, J.-C. and Zwickl, D.J. and Cho, S. and Davis, D.R. and Baixeras, J. and Brown, J. and Parr, C. and Weller, S. and Lees, D.C. and Mitter, K.T.} } @article {21183, title = {Body size affects the evolution of eyespots in caterpillars}, journal = {Proceedings of the National Academy of Sciences}, volume = {112}, year = {2015}, month = {05/2015}, pages = {6664-6669}, chapter = {6664}, abstract = {

\"Many caterpillars have conspicuous eye-like markings, called eyespots. Despite recent work demonstrating the efficacy of eyespots in deterring predator attack, a fundamental question remains: Given their protective benefits, why have eyespots not evolved in more caterpillars? Using a phylogenetically controlled analysis of hawkmoth caterpillars, we show that eyespots are associated with large body size. This relationship could arise because (i) large prey are innately conspicuous; (ii) large prey are more profitable, and thus face stronger selection to evolve such defenses; and/or (iii) eyespots are more effective on large-bodied prey. To evaluate these hypotheses, we exposed small and large caterpillar models with and without eyespots in a 2 \× 2 factorial design to avian predators in the field. Overall, eyespots increased prey mortality, but the effect was particularly marked in small prey, and eyespots decreased mortality of large prey in some microhabitats. We then exposed artificial prey to na\ïve domestic chicks in a laboratory setting following a 2 \× 3 design (small or large size \× no, small, or large eyespots). Predators attacked small prey with eyespots more quickly, but were more wary of large caterpillars with large eyespots than those without eyespots or with small eyespots. Taken together, these data suggest that eyespots are effective deterrents only when both prey and eyespots are large, and that innate aversion toward eyespots is conditional. We conclude that the distribution of eyespots in nature likely results from selection against eyespots in small caterpillars and selection for eyespots in large caterpillars (at least in some microhabitats).\"

}, keywords = {ANOVA, DEFENCE, EVOLUTION, EYESPOT, IMMATURE MORPHOLOGY, MIMICRY, PREDATION, SPHINGIDAE}, doi = {10.1073/pnas.1415121112}, author = {Hossie, T.J. and Skelhorn, J. and Breinholt, J.W. and Kawahara, A.Y. and Sherratt, T.N.} } @article {21182, title = {Quantifying wing shape and size of saturniid moths with geometric morphometrics}, journal = {Journal of the Lepidopterists{\textquoteright} Society}, volume = {70}, year = {2016}, pages = {99-107}, chapter = {99}, abstract = {

\"Butterflies and moths exhibit a spectacular diversity of wing shape and size. The extent of wing variation is particularly evident in wild silk moths (Saturniidae), which have large wing shape and size variation. Some species have jagged wing margins, rounded forewing apical lobes, or narrow hind wings with long tails, while others lack these traits entirely. Surprisingly, very little work has been done to formally quantify wing variation within the family. We analyzed the hind wing shape and size of 76 saturniid species representing 52 genera across five subfamilies using geometric morphometrics. We identified fifteen landmarks that we predict can be applied to families across Lepidoptera. PCA analyses grouped saturniid hind wings into six distinct morphological clusters. These groups did not appear to follow species relatedness\—some phylogenetically and genetically distantly related taxa clustered in the same morphological group. We discuss\  ecological factors that might have led to the extraordinary wing variation within Saturniidae.\"

}, keywords = {GEOMETRIC MORPHOMETRICS, LANDMARKS, MORPHOMETRICS, PRINCIPAL COMPONENTS ANALYSIS, PROCRUSTES ANALYSIS, SATURNIIDAE, WING LENGTH, WING SHAPE}, author = {Zhong, M. and Hill, G.A. and Gomez, J.P. and Plotkin, D. and Barber, J.R. and Kawahara, A.Y.} } @article {21181, title = {Wolbachia in butterflies and moths: geographic structure in infection frequency}, journal = {Frontiers in Zoology}, volume = {12}, year = {2015}, pages = {16}, chapter = {16}, abstract = {

\"Introduction: Butterflies and moths (Lepidoptera) constitute one of the most diverse insect orders, and play an important role in ecosystem function. However, little is known in terms of their bacterial communities. Wolbachia, perhaps the most common and widespread intracellular bacterium on Earth, can manipulate the physiology and reproduction of its hosts, and is transmitted vertically from mother to offspring, or sometimes horizontally between species. While its role in some hosts has been studied extensively, its incidence across Lepidoptera is poorly understood. A recent analysis using a beta-binomial model to infer the between-species distribution of prevalence estimated that approximately 40 \% of arthropod species are infected with Wolbachia, but particular taxonomic groups and ecological niches seem to display substantially higher or lower incidences. In this study, we took an initial step and applied a similar, maximum likelihood approach to 300 species of Lepidoptera (7604 individuals from 660 populations) belonging to 17 families and 10 superfamilies, and sampled from 36 countries, representing all continents excluding Antarctica.

Results: Approximately a quarter to a third of individuals appear to be infected with Wolbachia, and around 80 \% of Lepidoptera species are infected at a non-negligible frequency. This incidence estimate is very high compared to arthropods in general. Wolbachia infection in Lepidoptera is shown to vary between families, but there is no evidence for closely related groups to show similar infection levels. True butterflies (Papilionoidea) are overrepresented in our
data, however, our estimates show this group can be taken as a representative for the other major lepidopteran superfamilies. We also show substantial variation in infection level according to geography \– closer locations tend to show similar infection levels. We further show that variation in geography is due to a latitudinal gradient in Wolbachia infection, with lower frequencies towards higher latitudes.

Conclusions: Our comprehensive survey of Wolbachia infection in Lepidoptera suggests that infection incidence is very high, and provides evidence that climate and geography are strong predictors of infection frequency.\"

}, keywords = {CRAMBIDAE, DNA, DREPANIDAE, EPICOPEIDAE, EREBIDAE, EUPTEROTIDAE, GELECHIIDAE, GEOMETRIDAE, GRACILLARIIDAE, HESPERIIDAE, LASIOCAMPIDAE, LATITUDINAL GRADIENT, LYCAENIDAE, NOCTUIDAE, NYMPHALIDAE, PAPILIONIDAE, PIERIDAE, PLUTELLIDAE, PYRALIDAE, TORTRICIDAE, WOLBACHIA}, doi = {10.1186/s12983-015-0107-z}, author = {Ahmed, M.Z. and Araujo-Jnr, E.V. and Welch, J.J. and Kawahara, A.Y.} } @article {20925, title = {Tempo and mode of antibat ultrasound production and sonar jamming in the diverse hawkmoth radiation}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {112}, year = {2015}, month = {05/2015}, pages = {6407-6412}, chapter = {6407}, keywords = {ACOUSTICS, AMBULYX, ANCESTRAL STATE RECONSTRUCTION, BATS, BAYESIAN INFERENCE, CECHETRA, DEFENCE, DIVERGENCE TIMES, DNA, EPTESICUS, HYLES, JAMMING, MAXIMUM LIKELIHOOD, PHYLOGENY, PREDATION, PREDATOR AVOIDANCE, PSILOGRAMMA, RAXML, SONOGRAM, SPHINGIDAE, STRIDULATION, ULTRASONICS, XYLOPHANES}, url = {https://www.pnas.org/doi/full/10.1073/pnas.1416679112}, author = {Kawahara, A.Y. and Barber, J.R.} } @article {20886, title = {A molecular phylogeny of Eumorpha (Lepidoptera: Sphingidae) and the evolution of anti-predator larval eyespots}, journal = {Systematic Entomology}, volume = {40}, year = {2015}, pages = {401-408}, chapter = {401}, keywords = {BAYESIAN INFERENCE, BEHAVIOUR, BIOGEOGRAPHY, CAD, COI, DEFENCE, DNA, EF-1ALPHA, EUMORPHA, EYESPOT, IMMATURE STAGES, MAXIMUM LIKELIHOOD, MIMICRY, PHYLOGENY, RASP, RAXML, SPHINGIDAE, WINGLESS, WORLDMAP1}, doi = {10.1111/syen.12111}, url = {https://doi.org/10.1111/syen.12111}, author = {Ponce, F.V. and Breinholt, J.W. and Hossie, T. and Barber, J.R. and Janzen, D.H. and Hallwachs, W. and Kawahara, A.Y.} } @article {19854, title = {Phylogenetics and species status of Hawai{\textquoteleft}i{\textquoteright}s endangered Blackburn{\textquoteright}s Sphinx Moth, Manduca blackburni (Lepidoptera: Sphingidae)}, journal = {Pacific Science}, volume = {66}, year = {2012}, month = {2012}, pages = {31-41}, chapter = {31}, keywords = {BARCODING, CAD, COI, EF-1ALPHA, HAWAII, MANDUCA, PHYLOGENY, SPHINGIDAE, USA, WORLDMAP1}, doi = {10.2984/66.1.2}, author = {Rubinoff, D. and San Jose, M. and Kawahara, A.Y.} } @article {19853, title = {Synonomization [sic] of the euphonious Arctonotus Boisduval, 1852 (Sphingidae: Macroglossinae) based on molecular phylogenetic analysis}, journal = {Journal of the Lepidopterists{\textquoteright} Society}, volume = {63}, year = {2009}, month = {2009}, pages = {233-235}, chapter = {233}, keywords = {ARCTONOTUS, CATALOGUE, COI, EF-1ALPHA, PHYLOGENY, PROSERPINUS, SPHINGIDAE, WINGLESS}, url = {https://images.peabody.yale.edu/lepsoc/jls/2000s/2009/2009-63-4-233.pdf}, author = {Rubinoff, D. and Osborne, K.H. and Kawahara, A.Y.} } @article {18694, title = {Evolution of Manduca sexta hornworms and relatives: biogeographical analysis reveals an ancestral diversification in Central America}, journal = {Molecular Phylogenetics and Evolution}, volume = {68}, year = {2013}, month = {2013}, pages = {381-386}, chapter = {381}, keywords = {BAYESIAN INFERENCE, BIOGEOGRAPHY, CAD, COI, DNA, EF-1ALPHA, MANDUCA, MAXIMUM LIKELIHOOD, PEST, RASP, SPHINGIDAE, WINGLESS}, doi = {10.1016/j.ympev.2013.04.017}, url = {https://doi.org/10.1016/j.ympev.2013.04.017}, author = {Kawahara, A.Y. and Breinholt, J.W. and Ponce, F.V. and Haxaire, J. and Lei, X. and Lamarre, G.P.A. and Rubinoff, D. and Kitching, I.J.} } @article {17015, title = {Phylotranscriptomics: saturated third codon positions radically influence the estimation of trees based on next-gen data}, journal = {Genome Biology and Evolution}, volume = {5}, year = {2013}, month = {10/2013}, pages = {2082-2092}, chapter = {2082}, abstract = {

\"Recent advancements in molecular sequencing techniques have led to a surge in the number of phylogenetic studies that incorporate large amounts of genetic data. We test the assumption that analyzing large number of genes will lead to improvements in tree resolution and branch support using moths in the superfamily Bombycoidea, a group with some interfamilial relationships that have been difficult to resolve. Specifically, we use a next-gen data set that included 19 taxa and 938 genes (\∼1.2M bp) to examine how codon position and saturation might influence resolution and node support among three key families. Maximum likelihood, parsimony, and species tree analysis using gene tree parsimony, on different nucleotide and amino acid data sets, resulted in largely congruent topologies with high bootstrap support compared with prior studies that included fewer loci. However, for a few shallow nodes, nucleotide and amino acid data provided high support for conflicting relationships. The third codon position was saturated and phylogenetic analysis of this position alone supported a completely different, potentially misleading sister group relationship. We used the program RADICAL to assess the number of genes needed to fix some of these difficult nodes. One such node originally needed a total of 850 genes but only required 250 when synonymous signal was removed. Our study shows that, in order to effectively use next-gen data to correctly resolve difficult phylogenetic relationships, it is necessary to assess the effects of synonymous substitutions and third codon positions.\"

}, keywords = {ACTIAS, ANTHERAEA, BOMBYCIDAE, BOMBYX, CERATOMIA, DARAPSA, DNA, ENYO, HEMARIS, INCOMPLETE, MANDUCA, MAXIMUM LIKELIHOOD, PARSIMONY, PHYLOGENY, RADICAL, RAXML, SAMIA, SATURATION, SATURNIIDAE, SPHINGIDAE, SYNONYMOUS SUBSTITUTIONS, TRANSCRIPTOMICS}, doi = {10.1093/gbe/evt157}, url = {https://doi.org/10.1093/gbe/evt157}, author = {Breinholt, J.W. and Kawahara, A.Y.} } @article {16773, title = {Hawkmoths produce anti-bat ultrasound}, journal = {Biology Letters}, volume = {9}, year = {2013}, month = {2013}, pages = {20130161}, chapter = {20130161}, keywords = {BATS, DEFENCE, MALE GENITALIA, SPHINGIDAE, STRIDULATION, STRIDULATORY SCALES, ULTRASONICS}, doi = {10.1098/rsbl.2013.0161}, author = {Barber, J.R. and Kawahara, A.Y.} }