Use of mt COI sequences for the identification of subterranean termite species from Punjab, Pakistan

Correct species identification of termites is vital for their management and control. Species level identification becomes a difficult task due to biology, caste system and sexual dimorphism in subterranean termites. Worker and soldier caste termites were collected from various localities of Punjab and species were identified using morphometric characters of the soldier caste. Mitochondrial, COI-5ʹ (barcode region) and COI-3ʹ genes were sequenced from the identified soldier specimens and unidentified worker specimens to determine their molecular identities and phylogenetic relations. Both the DNA barcodes and COI-3ʹ sequences discriminated the analyzed termites to the species level. The cluster analysis showed species level relationships between different termite species. Disparity Index Test of species based on barcode data showed significant differences among majority of the species pairs. Nucleotide similarity analysis of the species showed that none of the species had a similarity of more than 98% and in most cases similarity was less than 89%. Analysis of COI-3ʹ sequences of seven termite species when performed along with species sequences reported in the GenBank placed the species of each genus together in a single cluster. In conclusion, barcode and COI-3ʹ regions revealed nucleotide differences among termite species which can be used to discriminate among termite species.