Molecular phylogenetic analysis of the subfamily Chilocorinae (Coleoptera: Coccinellidae) from the Malakand region, Pakistan

The subfamily Chilocorinae plays an active role in biological control, being predominantly predatory in nature and distributed worldwide, including in Pakistan. They feed on soft-bodied insects such as scale insects, aphids, leafhoppers, mealybugs, and psyllids. In this research, three species have been molecularly identified for the first time, belonging to three different genera: Brumoides, Exochomus, and Parexochomus. The study was conducted from April 2022 to April 2023, during which a total of 375 samples were collected from the Malakand region of Pakistan, specifically from seven districts. The collected specimens were preserved in insect boxes and tagged. DNA extraction was performed from the legs of ladybird beetles, followed by PCR amplification using the primers LCO1490 and HCO2198. Sanger sequencing was carried out using BioEdit software (version 7.2), and phylogenetic analyses were performed using MEGA 11 software to construct a phylogenetic tree. Three species belonging to three genera of the subfamily Chilocorinae were molecularly identified and phylogenetically analyzed from the Malakand region of Pakistan: Brumoides suturalis (Fabricius, 1789), Exochomus nigripennis (Erichson, 1843), and Parexochomus nigromaculatus (Goeze, 1777). Overall, this study underscores the importance of phylogenetic analysis for the accurate taxonomy of the subfamily Chilocorinae and their relationship with closely related species.