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The South American leafminer, Tuta absoluta is an exotic devastative pest on solanaceous vegetables, including tomatoes, which leads to a cent per cent economic loss in India. The molecular markers assist in assessing gene flow, migratory frequencies, and genetic variety, as well as helping to evaluate the genetic makeup and diversification of an exotic species population to indigenous ones. With this, the present study aimed to investigate the genetic divergence of T. absoluta in different districts of Tamil Nadu, India. The study depicted the examination of genetic divergence of T. absoluta by aiding amplified region of mitochondrial DNA encoding cytochrome oxidase I (COI) from the T. absoluta samples gathered from Coimbatore, Dharmapuri and Dindigul districts of Tamil Nadu. The findings showed that the phylogenetic tree constructed from all sequences of T. absoluta acquired from the NCBI (National Center for Biotechnology Information) and BOLD (The Barcode of Life Data System) databases exhibited 99 percent identity and aggregated together into a single clade. Hence, the present study revealed the great genetic uniformity in T. absoluta populations in India and corroborates that most of the globe rely on the partial COI gene, evidenced by minimal nucleotide diversity. [ABSTRACT FROM AUTHOR]