The Complete Mitochondrial Genomes of Four Species in the Subfamily Limenitidinae (Lepidoptera, Nymphalidae) and a Phylogenetic Analysis

SIMPLE SUMMARY: As it is currently delineated, the subfamily Limenitidinae (Lepidoptera: Nymphalidae) is comprised of 50 genera with approximately 1100 species. The classification of this subfamily has always been unstable. There are tribes and genera whose status is doubtful. Their phylogenetic relationships are far from being clarified and the monophyly of some of them is under question. To provide further insight into the relationships among included tribes, four newly-completed mitochondrial genomes of Limenitidinae (Neptis thisbe, Athyma zeroca, and Aldania raddei) have been sequenced and analyzed. Results indicate that the gene orientation and arrangement are similar to typical mitogenomes in Lepidoptera. The inferred phylogenetic analysis shows that tribe levels are well-supported monophyletic groups. Taken together, this work will provide a well-resolved framework for future study of this subfamily. ABSTRACT: The complete mitogenomes of four species, Neptis thisbe, Neptis obscurior, Athyma zeroca, and Aldania raddei, were sequenced with sizes ranging from 15,172 bp (N. obscurior) to 16,348 bp (Al. raddei). All four mitogenomes display similar nucleotide content and codon usage of protein-coding genes (PCGs). Typical cloverleaf secondary structures are identified in 21 tRNA genes, while trnS1 (AGN) lacks the dihydrouridine (DHC) arm. The gene orientation and arrangement of the four mitogenomes are similar to that of other typical mitogenomes of Lepidoptera. The Ka/Ks ratio of 13 PCGs among 58 Limenitidinae species reveals that cox1 had the slowest evolutionary rate, while atp8 and nad6 exhibited a higher evolutionary rate. The phylogenetic analysis reveals that tribe-levels are well-supported monophyletic groups. Additionally, Maximum Likelihood analysis recovered the relationship (Parthenini + ((Chalingini + (Cymothoini + Neptini)) + (Adoliadini + Limenitidini))). However, a Bayesian analysis based on the same dataset recovered the relationship (Parthenini + (Adoliadini + ((Cymothoini + Neptini) + (Chalingini + Limenitidini)))). These results will offer valuable data for the future study of the phylogenetic relationships for Limenitidinae.