Six species of Nymphalidae butterflies in habiting in Mukurthi National Park (The Nilgiri Biosphere Reserve, Western Ghats, India) were identified through DNA barcoding based on 650bp sequences of the mitochondrial COI gene. The sequences of these native species were compared with data bases available for these six species in India and other countries to improve the understanding of inter and intra species divergence. Neighbour-Joining cluster analysis with one out group, Pachilopta hector, and three different haplotypes revealed pair-wise distance comparison with the nearest neighboring species in BOLD database showed overall nucleotide divergent between 0.01-0.19%. The mean inter and intra specific nucleotide divergence of these six species was calculated to be 0.00827. As per the 10X rule, the threshold value was 0.08%. Therefore, no deep intra species nucleotide divergence was observed and no significant barcoding gap existed between the selected species. In this study, the native species in each haplotype was found to be very close relative to non-native species. The overlapping of the DNA sequences occurs due to less variation.
