Abstract
Anopheles mosquitoes serve as a significant vector for the transmission of malaria and arboviruses holding great medical importance, but their phylogenetic relationships have not yet been clearly elucidated. This study was the first to sequence and analyze the mitochondrial genomic features of An. hyrcanus and An. messeae, and to incorporate them into a phylogenetic analysis with 102 other species within the genus Anopheles. Both genomes contain 37 genes, including 22 tRNAs, 13 protein-coding genes and 2 rRNAs, as well as one control region. The genome characteristics, codon usage, and tRNA secondary structure of the two species align with those observed in mitochondrial genomes of other mosquito species. Phylogenetic analysis revealed that the relationship among six subgenera was Lophopodomyia + ((Kerteszia + Stethomyia) + ((Cellia + Anopheles) + Nyssorhynchus)), providing strong support for their monophyly. Nevertheless, the phylogenetic relationships within subgenus Cellia were inconsistent with previous findings. The findings of this study will enrich the biological information repository for Anopheles mosquitoes, lay the foundation for further research on systematic classification, and hold significant implications for guiding the research, prevention and control of mosquito-borne diseases.