Evaluating the DNA barcodes for identification of butterflies from Western Himalayas,Uttarakhand, India

DNA barcodes in species tagging have become a popular tool for taking inventories of species from different groups worldwide. The present study aimed to generate DNA barcodes of butterfly species from the Western Himalayas in Uttarakhand, India. The Indian Western Himalayan region (IWHR) has been explored to a limited extent about butterfly species’ diversity. However, the IWHR is prone to environmental change, and slight variations in climatic conditions can influence species diversity and change butterflies' range. The mitochondrial cytochrome c oxidase I (COI) gene was first used to generate the DNA barcode for butterflies from this region on a broad scale. 28 morphologically identified species, consisting of 102 sequences, were finally grouped into 26 species, with only two species showing ambiguity in species identification. These species had < 3% sequence variations from their neighboring relatives, suggesting cryptic species diversity. Generated sequences were also compared with the GenBank data of conspecific geographical locations, which showed intraspecies variation ranging from 1.3% to 7.3%. It was also noted that butterfly species have both intra and interspecies sequence divergence. In the phylogenetic-based species identification, a total of 28 species belonging to 4 families of butterflies were successfully discriminated, and two species at the genus level, which had high intra-specific divergence (0.025), were considered. However, the high intra-species sequence divergence observed may represent the presence of hidden species.