Phylogenetic relationship and morphological evolution in the subfamily Limenitidinae (Lepidoptera: Nymphalidae)

Abstract

The mitochondrial cytochrome oxidase subunit I (COI) gene and the nuclear elongation factor 1α (EF-1α) gene were sequenced from 29 species of Nymphalidae (Nymphalidae, Lepidoptera). Phylogenetic trees were constructed based on the sequences determined from the 29 species and sequences of other 36 species deposited in GenBank using the neighbor-joining (NJ), maximum likelihood (ML) and Bayesian methods with Libythea celtis (Libytheinae) as the outgroup. Our phylogenetic trees indicated four major clades. Clade A includes three subfamilies: Apaturinae, Nymphalinae, and Limenitidinae, excluding the tribe Limenitidini; Clade B includes the subfamilies Heliconiinae and the tribe Limenitidini; Clade C includes Satyrinae, Calinaginae, Charaxinae and Morphinae; and Clade D includes subfamily Danainae. Our study suggested that the tribes Pseudergolini, Biblidini, Limenitidini and Cyrestidini should be considered as subfamilies and confirmed the interspecific relationships within the subfamily Pseudergolinae, namely Amnosia + (Pseudergolis + (Stibochiona + Dichorragia)). We then mapped three morphological characters (spot of anal angle, eyespots, and process from outer margin of hind wing) onto the phylogenetic tree constructed by ML analysis using the combined sequence data. Based on this the evolutionary patterns of these morphological characters were identified, they indicated that the three characters evolved repeatedly in the family Nymphalidae.