Four complete mitochondrial genomes of the subgenus (Diptera, Tipulidae, ) and implications for the higher phylogeny of the family Tipulidae.

The complete mitochondrial genomes of Tipula (Pterelachisus) cinereocincta mesacantha Alexander, 1934, T. (P.) legalis Alexander, 1933, T. (P.) varipennis Meigen, 1818, and T. (P.) yasumatsuana Alexander, 1954 are reported, three of them being sequenced for the first time. The mitochondrial genome lengths of the four species are 15,907 bp, 15,625 bp, 15,772 bp, and 15,735 bp, respectively. All genomes exhibit a high AT base composition, with A + T content of 76.7%, 75.0%, 77.8%, and 75.4%, respectively. The newly reported mitogenomes herein show a general similarity in overall structure, gene order, base composition, and nucleotide content to those of the previously studied species within the family Tipulidae. Phylogenetic analyses were conducted to investigate the relationships within Tipulidae, using both Maximum Likelihood and Bayesian Inference approaches. The results show that the four target species of the subgenus T. (Pterelachisus) basically form a monophyletic group within Tipulidae, clustering with species of the subgenera T. (Lunatipula), T. (Vestiplex), and T. (Formotipula); however, the genus is not monophyletic. Moreover, neither the tipulid subfamily Tipulinae nor the family Limoniidae is supported to be a monophyletic group. The monophyly of the family Tipulidae, and the sister relationship between Tipulidae and Cylindrotomidae are reconfirmed. These research findings could contribute to deep insights into the systematic and evolutionary patterns of crane flies.