To confirm the pathogenicity differentiation of pathogens of tobacco bushy top disease from different geographical regions and tobacco hosts, twelve wild pathogen samples were collected, the pathogens were purified and their pathogenicity was determined in greenhouse with aphid-vector and rubbing transmission method respectively. The disease incidence of pathogen TBTV-7 from Menghai County in Xishuangbanna were 17% and 16% respectively on the 28th day after inoculated by the two methods, and that of other pathogens were over 70%. It indicated the existence of pathogenicity differentiation.