Abstract: Random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) was used to survey the B biotype and other biotypes of Bemisia tabaci in Jordan. Whiteflies were collected from cultivated plants, wild plants and weeds from (9) different localities and on (12) distinct crops with total (123) whitefly insect. RAPD analyses using three selected 10-mer primers reliably identified B. tabaci biotypes. The total number of clear bands obtained from it was 29 DNA bands. Cluster analysis demonstrated that, in general, biotype B individuals which are scattered independently in the localities according to the host plant and coexists with cultivated plants. While an intermediate whitefly populations BA having biotype A a distinguish band only with OPR-04 but according to Jaccard similarity they have higher genetic distance with biotype B and this samples where restricted to wild plants and weeds. And finally the biotype A was restricted to the isolated area comported to other biotype. The percentage of Whitefly biotype A, B and BA in the sample was approximately 12.5, 75 and 12.5% Jordan Valley and Upland of tested samples.