Abstract

BackgroundSequence divergence in mitochondrial cytochrome c oxidase subunit I genes (COI) has been used as an efficient forensic tool in solving wildlife-related problems and also be used in molecular taxonomy for species identification.MethodsThis study presents the DNA barcode sequences of the Blackbuck (Antilope cervicapra) from Haryana, India. A dataset of 43 partial COI sequences of 13 species belonging to 4 genus of Antilopinae sub- family were used for molecular analysis and to construct the phylogenetic tree to elucidate the species diversity among sub-family Anitlopinae. The data comprised of generated sequences of the Blackbuck (n = 22) and with additional COI sequences of the related species (n = 21) showing maximum homology with Blackbuck COI sequence were downloaded from NCBI-GenBank for wide coverage of inter and intra-specific nucleotide diversity.ResultsPhylogenetic analysis demonstrated that the studied COI region provided accurate species clustering showing their importance in wildlife species identification. The intra-specific sequence divergence of all the studied Antilopinae species was observed 0.47% (<1%). Specifically in the studied genus- Antilope, it was observed 0.3%. Whereas the interspecific divergence was in the range of 0.3–12.6% among 13 species from 4 genus. The highest interspecific divergence was observed among Antilope cervicapra and Gazella erlangeri (12.6%).ConclusionThe developed species-specific barcoding (COI) sequence of Indian Blackbuck from Haryana, India, demonstrated their high potential to identify Blackbuck from their sympatric species in wildlife-related crimes and in conservation activities. Published Barcode sequences in the both database serve as a wildlife forensic tool to solve Blackbuck related crimes. This study supports the credibility of DNA Barcoding in forensic investigation and wildlife conservation and also emphasized the need for species barcode database from each country to assist in solving wildlife related offenses.

Highlights

  • Sequence divergence in mitochondrial cytochrome c oxidase subunit I genes (COI) has been used as an efficient forensic tool in solving wildlife-related problems and be used in molecular taxonomy for species identification

  • Primer selection A universal set of Primer was selected for this study from the primer database available in Barcode of Life Database (BOLD) system to target the barcode COI sequence of the Antilope cervicapra

  • The data comprised of generated sequences of the Blackbuck (n = 22) and with additional COI sequences of the related species (n = 21) showing maximum homology were used and downloaded from National Center for Biotechnology Information (NCBI)-GenBank for wide coverage of inter and intra-specific nucleotide diversity (Table 1)

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Summary

Introduction

Sequence divergence in mitochondrial cytochrome c oxidase subunit I genes (COI) has been used as an efficient forensic tool in solving wildlife-related problems and be used in molecular taxonomy for species identification. A DNA barcode is a standardized approach used to identify species from partial sequences of DNA (Hebert and Gregory, 2005; Kumar et al 2012) This technique uses the mitochondrial region of cytochrome C oxidase subunit I (COI) to provide a strong species-level resolution for diverse wildlife (Linacre & Tobe 2011). The DNA barcode region contains highly conserved sites for primer binding, and it can be used to show interspecific variations (Ferri et al 2009; Eaton et al 2010). This method has been attracting international attention due to its significant role in advancing the taxonomy of life forms and their forensic implications

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