Abstract
BackgroundButterflies (Papilionoidea) are perhaps the most charismatic insect lineage, yet phylogenetic relationships among them remain incompletely studied and controversial. This is especially true for skippers (Hesperiidae), one of the most species-rich and poorly studied butterfly families.MethodsTo infer a robust phylogenomic hypothesis for Hesperiidae, we sequenced nearly 400 loci using Anchored Hybrid Enrichment and sampled all tribes and more than 120 genera of skippers. Molecular datasets were analyzed using maximum-likelihood, parsimony and coalescent multi-species phylogenetic methods.ResultsAll analyses converged on a novel, robust phylogenetic hypothesis for skippers. Different optimality criteria and methodologies recovered almost identical phylogenetic trees with strong nodal support at nearly all nodes and all taxonomic levels. Our results support Coeliadinae as the sister group to the remaining skippers, the monotypic Euschemoninae as the sister group to all other subfamilies but Coeliadinae, and the monophyly of Eudaminae plus Pyrginae. Within Pyrginae, Celaenorrhinini and Tagiadini are sister groups, the Neotropical firetips, Pyrrhopygini, are sister to all other tribes but Celaenorrhinini and Tagiadini. Achlyodini is recovered as the sister group to Carcharodini, and Erynnini as sister group to Pyrgini. Within the grass skippers (Hesperiinae), there is strong support for the monophyly of Aeromachini plus remaining Hesperiinae. The giant skippers (Agathymus and Megathymus) once classified as a subfamily, are recovered as monophyletic with strong support, but are deeply nested within Hesperiinae.ConclusionsAnchored Hybrid Enrichment sequencing resulted in a large amount of data that built the foundation for a new, robust evolutionary tree of skippers. The newly inferred phylogenetic tree resolves long-standing systematic issues and changes our understanding of the skipper tree of life. These resultsenhance understanding of the evolution of one of the most species-rich butterfly families.
Highlights
Butterflies (Papilionoidea) are perhaps the most charismatic insect lineage, yet phylogenetic relationships among them remain incompletely studied and controversial
All extracts are stored at − 80 °C at the McGuire Center for Lepidoptera and Biodiversity (MGCL), Florida Museum of Natural History (Gainesville, USA)
Two species obtained via published transcriptomes had < 300 loci assembled; the skipper Erynnis propertius (Pyrginae, 155 loci; [59]), and the outgroup Macrosoma hedylaria, (Hedylidae, 175 loci; [40])
Summary
Butterflies (Papilionoidea) are perhaps the most charismatic insect lineage, yet phylogenetic relationships among them remain incompletely studied and controversial. This is especially true for skippers (Hesperiidae), one of the most species-rich and poorly studied butterfly families. Using a small number of gene fragments sampled across the genome, phylogenetic relationships of diverse groups have been inferred and their taxonomy revised. Genomic regions that hybridize with probes are enriched with PCR and can be sequenced. AHE can work well for degraded DNA from older specimens [4], permitting the use of specimens with DNA too fragmented for PCR or reduced representation methods requiring restriction enzymatic cleavage. The use of AHE in systematics is still in its infancy, but pioneering studies have shown the astonishing potential of this method to fully resolve deeper parts of the ToL
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