Genetic Differentiation and Phylogenetic Analysis of Belenois Butterfly Species (Belenois aurota and Belenois java) (Pieridae) Using DNA Barcoding of the COI Gene from A.P. – India

This study focuses on the molecular identification and genetic differentiation of two widely distributed 2 distinct Belenois butterfly species, Belenois aurota (Fabricius 1973) and Belenois java (Sparman 1968), through DNA barcoding of the mitochondrial COI gene from species collected from Seshachalam Bio reserve, A.P. Eggs were collected from Capparis zeylanica host plants and specimens were identified morphologically before DNA extraction. PCR amplification and sequencing of the COI gene revealed clear genetic distinctions between the two species, with sequence lengths of approximately 701 bp and 703 bp. Phylogenetic analysis and BLAST searches confirmed species identity and highlighted the utility of DNA barcoding in resolving species and assessing biodiversity. The results underscore the importance of molecular techniques in butterfly taxonomy, conservation, and ecological studies, providing a valuable reference for future biodiversity monitoring efforts in the region. The Barcode of Life Database (BOLD, www.barcodinglife.org) was updated with the sequence information, picture, and supporting documentation for every voucher specimen. Sequencing-Systematics of certain tropical Butterflies in and around Southern Andhra Pradesh [KSVGP] is the project file that contains the BOLD's detailed specimen records and sequencing information, including trace files.