Bemisia tabaci has been well documented as one of the most economically important emergent plant virus vectors, through serious feeding damage to its broad range of plant hosts to important agricultural crops. Among the crops cotton is an important cash crop of Pakistan. Pakistan is the fourth largest cotton producer country in the world. It holds importance to run the huge textile industry in Pakistan. Production of good quality cotton is affected by Cotton Leaf Curl disease in cotton producing areas of Pakistan, B. tabaci is an efficient vector of CLCuV, its ability to transmit disease varies with its genetic composition that may be determined by geography and climatic conditions of that location. There is an urgent need to unfold the secrets behind its diversity, but efforts to explore the genetic status of this insect were insufficient and inconsistent and multiple approaches are available to identify the species/biotype status of B. tabaci further add confusion to the situation. To evaluate the biotype of B. tabaci specific to this region was important. The purpose of this study was to provide information about the biotype and secondary endosymbiont distribution for B. tabaci populations in the Punjab. There are two parts of this study, the first part deals with the exploration of biotypes, the genetic and haplotype diversity among the COI sequences of B. tabaci of Punjab. The second part deals with the identification of 16S rDNA sequences of endosymbionts of B. tabaci, and its phylogenetic analysis. The geographical localization and variation in endosymbiont populations across the region were identified using a sequence-driven analysis of the population genetics of the secondary endosymbiont. Live field specimens were collected from 39 different locations among 16 districts of Punjab and preserved in Absolute ethanol for genetic studies. Previously established procedures were used to extract and purify total insect DNA from 3 individual whiteflies for each location. PCR was run with two sets of primers designed on two loci for the highly conserved whitefly Mitochondrial Cytochrome Oxidase subunit I (COI) gene for biotyping. The Bayesian phylogeny of COI sequences revealed the B. tabaci populations of Punjab belongs to Asia I genetic group. Out of all the 7 genetic clades of B. tabaci, it was observed that sequences of COI of this study grouped with the sequences from the neighbouring countries like China, India and Nepal in Asia I clade. Specimens were subjected to PCR amplification using the general 16S rDNA primers for the Portiera, Arsenophonus, Rickettsia, Cardinium and Wolbachia to amplify endosymbiont DNA. The ML trees of endosymbionts of B. tabaci showed that the endosymbionts strains detected in this study grouped with the previously reported endosymbionts and these endosymbionts found to be the other strains of known endosymbionts of B. tabaci from other parts of the world. Samples were sequenced using the Sanger method and the data analyzed to correlate the presence, prevalence and geographical distribution of endosymbionts in B. tabaci. Phylogenies were constructed to track evolutionary differences amongst the endosymbionts and insects and how they have influenced the evolution of the regional populations. Samples were characterized by differences in the genomes and endosymbionts distribution among the population of B. tabaci from Punjab.