Molecular Identification and Phylogenetic Analysis of the Indian Leaf-Nosed Bat (Hipposideros speoris) from Tamil Nadu, India
Abstract
Accurate species identification is essential for understanding biodiversity, evolutionary relationships, and conservation of bats. However, morphological similarities among closely related species often limit traditional taxonomic approaches. The present study employed mitochondrial cytochrome c oxidase subunit I (COI) gene sequencing to investigate the molecular identity and phylogenetic relationship of the Indian leaf-nosed bat, Hipposideros speoris, from Tirunelveli District, Tamil Nadu, India. Genomic DNA was extracted from wing membrane tissue samples using the phenol–chloroform method, and a partial COI gene fragment of approximately 650 bp was amplified using universal Folmer primers. The amplified products were sequenced and compared with reference sequences available in the NCBI GenBank database. Phylogenetic analysis was performed using the Neighbour-Joining method based on Kimura 2-Parameter genetic distances with 1,000 bootstrap replications. The obtained COI sequence was successfully identified as H. speoris and clustered within the genus Hipposideros with strong bootstrap support. Genetic distance analysis revealed a divergence of 0.1638 between the present study sequence and the reference H. speoris sequence, while interspecific divergence with other Hipposideros species ranged from 0.1463 to 0.1891. The highest divergence was observed with the outgroup species Taphozous melanopogon (0.2477). The findings demonstrate the effectiveness of COI-based DNA barcoding for molecular identification and phylogenetic assessment of bats and provide valuable insights into the genetic diversity and evolutionary relationships of H. speoris.
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