Two Complete Mitochondrial Genomes of Mileewinae (Hemiptera: Cicadellidae) and a Phylogenetic Analysis

SIMPLE SUMMARY: Mileewinae is a small subfamily of Cicadellidae containing about 160 described species, extensively distributed in the Oriental, Ethiopian and Neotropical regions. Some species are potential pests in agriculture and forestry. The classification of this group has been unstable over the past few decades. Currently, some controversies remain on the monophyly of Mileewinae and phylogenetic relationships of Mileewinae with other subfamilies. To provide further evidence toward answering these questions, two newly completed mitochondrial genomes of Mileewinae species (Mileewa rufivena and Ujna puerana) have been sequenced and analyzed. Results show these two mitochondrial genomes have quite similar structures and features. In phylogenetic analyses, Mileewinae formed a monophyletic group in Cicadellidae in all trees derived from maximum likelihood (ML) and Bayesian inference (BI) methods. In addition, Mileewinae has a closer phylogenetic relationship with Typhlocybinae compared to the Cicadellinae. ABSTRACT: More studies are using mitochondrial genomes of insects to explore the sequence variability, evolutionary traits, monophyly of groups and phylogenetic relationships. Controversies remain on the classification of the Mileewinae and the phylogenetic relationships between Mileewinae and other subfamilies remain ambiguous. In this study, we present two newly completed mitogenomes of Mileewinae (Mileewa rufivena Cai and Kuoh 1997 and Ujna puerana Yang and Meng 2010) and conduct comparative mitogenomic analyses based on several different factors. These species have quite similar features, including their nucleotide content, codon usage of protein genes and the secondary structure of tRNA. Gene arrangement is identical and conserved, the same as the putative ancestral pattern of insects. All protein-coding genes of U. puerana began with the start codon ATN, while 5 Mileewa species had the abnormal initiation codon TTG in ND5 and ATP8. Moreover, M. rufivena had an intergenic spacer of 17 bp that could not be found in other mileewine species. Phylogenetic analysis based on three datasets (PCG123, PCG12 and AA) with two methods (maximum likelihood and Bayesian inference) recovered the Mileewinae as a monophyletic group with strong support values. All results in our study indicate that Mileewinae has a closer phylogenetic relationship to Typhlocybinae compared to Cicadellinae. Additionally, six species within Mileewini revealed the relationship (U. puerana + (M. ponta + (M. rufivena + M. alara) + (M. albovittata + M. margheritae))) in most of our phylogenetic trees. These results contribute to the study of the taxonomic status and phylogenetic relationships of Mileewinae.