First COI barcode record and phylogenetic assessment of Pramadea lunalis (Guenée, 1854) from Kerala, Western Ghats, India

Abstract

DNA barcode data remain limited for many Indian Spilomelinae, making reliable species identification and regional comparisons difficult. Pramadea lunalis (Guenée, 1854), originally described from India, is often reported as Syllepte lunalis in older literature and public databases. This study generated and evaluated COI barcode data for P. lunalis from Kerala, India, in the Western Ghats. Two male specimens were identified using external morphology and examination of the male genitalia, with reference to published taxonomic descriptions. Genomic DNA was extracted from ethanol-preserved leg tissue. The mitochondrial cytochrome c oxidase subunit I (COI) region was amplified using the Folmer primers LCO1490 and HCO2198. The sequences were analyzed using BLASTn, pairwise genetic distances, and Neighbor-Joining (NJ) and Maximum Likelihood (ML) methods. The two Kerala sequences were deposited in GenBank under accession numbers PX071900 and PZ343591. Both showed 95.7% identity with available Southeast Asian S. lunalis sequences, with a genetic divergence of approximately 4.3–4.5%. The Kerala specimens formed a strongly supported clade in both NJ and ML analyses and were recovered as the sister group of the Southeast Asian S. lunalis sequences. The observed divergence may reflect geographic differentiation or lineage-level variation, although inconsistencies in database nomenclature may also influence sequence comparisons. This study provides the first COI barcode records of P. lunalis from India and expands the available molecular reference data for the species. Broader geographic sampling, detailed morphological comparisons, and additional nuclear markers are needed to clarify the taxonomic significance of the observed divergence.