DNA barcoding and phylogenetic analysis of the subfamily Epilachninae (Coleoptera: Coccinellidae) from the Malakand Region, Pakistan

Kausar Saeed, Munawar Saleem Ahmad, Khushi Muhammad, Qaiser Zaman, Azam Jan Afridi, Imad Ali Khan, Ubaid Ur Rehman, Usama Nawab, Falak Naz, Farrah Zaidi

Abstract


The taxonomic identification of the family Coccinellidae, including the Ladybird beetles, remains ambiguous due to considerable morphological variability. In recent years, molecular phylogenetic analysis and DNA barcoding have played a crucial role in validating species taxonomy. This study, therefore, aimed to investigate the DNA barcoding and phylogenetic relationships of the subfamily Epilachninae in the Malakand region of Khyber Pakhtunkhwa, Pakistan. The survey was conducted from April 2022 to April 2023. Morphological identification was carried out using various standard taxonomic keys. DNA extraction and PCR amplification were performed using the universal primers LCO1490 and HCO2198, followed by Sanger sequencing. Phylogenetic analysis was conducted using various bioinformatics tools. A total of 510 specimens belonging to the subfamily Epilachninae were collected, comprising four species: Henosepilachna vigintioctopunctata (Fabricius, 1755), H.septima (Dieke, 1947), Afidentula manderstjernae (Mulsant, 1853), and Subcoccinella vigintiquatuorpunctata, representing three genera within the subfamily. This study underscores the significance of molecular phylogenetics in resolving the taxonomy and elucidating the evolutionary relationships of Epilachninae species in the Malakand region.


Keywords


Family Coccinellidae, Subfamily Epilachninae, DNA barcoding, Molecular Phylogenetic analysis, Malakand, Pakistan

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References


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DOI: https://doi.org/10.33804/pp.009.03.5672

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