Phylogeny, divergence times, and biogeography of the subfamily Tagiadinae (Lepidoptera: Hesperiidae) based on molecular data and morphological characters

The skipper subfamily Tagiadinae has been a key group in taxonomic debates and phylogenetic inconsistencies due to limited taxon sampling and morphological evidence. In this study, we aimed to resolve intra-subfamilial relationships within Tagiadinae using 3 datasets: (i) a molecular dataset (3 genes COI, COII, EF-1 alpha) including 92 species from 30 genera across all the 3 tribes of Tagiadinae, (ii) a morphological dataset (62 morphological or biological traits) comprising 50 species from 20 genera, and (iii) the combined dataset, representing the intersection of the first 2, containing 46 species from 20 genera. Both the molecular tree and the combined molecular-morphological tree supported the monophyly of Tagiadinae and its internal tribes, and the tribe Netrocorynini was consistently recovered as the sister group of the other Tagiadinae species. In addition, divergence time estimation suggested a crown age of approximately 37 million years for Tagiadinae, with the 2 tribes Celaenorrhinini and Tagiadini having diverged approximately 36 million years ago. The morphological and molecular evidence presented in this study contributes to a more robust understanding of the evolutionary framework of Tagiadinae. Our findings challenge some previous phylogenetic hypotheses regarding the basal position of Netrocorynini within Tagiadinae and provide valuable insights into the taxonomy and phylogeny of skipper butterflies.