Molecular characterization of Aedes, Culex, Anopheles, and Armigeres vector mosquitoes inferred by mitochondrial cytochrome oxidase I gene sequence analysis

The current epidemics of vector-borne diseases in tropical countries, especially in India, have extended the need for a comprehensive understanding of the distribution of epidemiologically important vector mosquitoes. The advanced developments in genomic research including phylogenetic studies will empower the molecular studies that are vital to determine the genetic divergence and evolutionary history of mosquito vectors. The use of the cytochrome oxidase c subunit I (COI) gene-based approaches to outline the interrelationship of vector mosquitoes from different genera could also elucidate the obscurity that has risen from improper taxonomical classification. The mosquitoes were collected from Nelliyampathy, Kerala, India and identified using systematic keys and catalogues. The genomic DNA of the mosquitoes was extracted using NucleoSpin® Tissue Kit and PCR amplification of the mitochondrial COI gene was accomplished by using the following primers: Forward 5′- GGTCAACAAATCATAAAGATATTGG-3′ and reverse 5′- TAAACTTCAGGGTGACCAAAAAATCA-3′. The genetic divergence was assessed by means of the NJ-K2P method. The neighbor-joining tree was constructed using the MEGA7 software. The diversity of mosquitoes was estimated using the Shannon index through SPSS and Venn diagram plotter. The COI gene-based mitochondrial DNA analysis revealed distinct clustering of individual mosquito species within every genus together with strong bootstrap support. In total, our investigation productively identified and confirmed the interrelationship between the COI gene sequences of ten epidemiologically important mosquito species. This study also discusses the substantial evidence of mosquito species complex formation. Our findings offer a base for future research which is essential for the better understanding of mosquito phylogeny and shed light upon, a novel vector control strategy.