Wild pomegranate ( Punica granatum L.) is one of the most important medicinal wild fruit crop and it is highly pollinated by wild insect pollinators. So, study analyze various pollinating insects of Punica granatum L. by using molecular method and a total of 24 species of flower visitors were collected from Punica granatum from different areas of Himachal Pradesh. Using mtDNA marker, the cytochrome oxidase subunit sequence I (COI), about 17 pollinator insect species were characterized and identified on the basis of COI sequence. BLAST analysis showed the 98 to 100% similarity with the existing genebank sequences. Nucleotide composition revealed the average AT content was significantly higher by 72.56% than the GC content of 27.47%. Phylogenetic analysis of 17 different species of insect pollinators of Punica granatum revealed the close phylogenetic relation between hymenopteran insect species in one clade, whereas another clade showing relationship between Lepidoptera, Coleopteran and Diptera. The study conclude that molecular identification using species identification markers in the mitochondrial region of COI play a critical role in study of genetic diversity, genetic relatedness and phylogenetic analysis at species level and find more strategies to conserve these pollinators for further research.