Comparative mitochondrial genomics unveils the phylogenetic relationships of lepidopteran moths

Moths constitute the largest group within Lepidoptera and play a crucial role in natural ecosystems. However, the current high-level classification system of Lepidopteran insects necessitates systematic research supported by comprehensive data. In this study, we determined the complete mitochondrial genomes of 80 lepidopteran moth species, spanning 68 genera across 13 families. Through a synthesis of published data, we conducted a comparative analysis encompassing 211 moth species from 12 superfamilies. Our analysis unveiled a mitochondrial genome length range of 15,027 to 17,049 bp, with an AT content varying between 77.02% and 83.52%. While gene composition and arrangement were largely conserved, we observed tRNA rearrangement (trnS1-trnE inversion) in Zygaenidae and Gelechiidae. Phylogenetic analyses highlighted the monophyly of 9 superfamilies, with exceptions for Tineoidea, Pyraloidea, and Drepanoidea. Divergence time estimations suggest an origin of the 12 Lepidopteran superfamilies in the mid-Cretaceous period, approximately 122.27 million years ago (95% CI: 102.43-153.23 Mya). Furthermore, we propose reclassifications to refine the taxonomic status of certain families. This includes reclassifying Gracillariidae and Thyrididae as independent from the Tineoidea and Pyraloidea superfamilies, respectively, and relocating the family Epicopeiidae to the Geometroidea superfamily from Drepanoidea. Overall, this study contributes valuable insights and resources to the systematic evolution of Lepidoptera.