Abstract: To explore biocontrol resources against tobacco bacteria wilt, a plate dilution method was used to isolate endophytic bacteria from healthy tobacco plants and rhizosphere bacteria from rhizosphere soils in four tobacco-growing areas of Qiannan in Guizhou Province. The results showed that a total of 103 endophytic bacteria and 73 rhizosphere bacteria were obtained. Endophytic bacteria LSN02 and rhizosphere bacteria LLGJ04 against tobacco bacteria wilt were screened by inhibition zone method, and their inhibition zone diameter were 29.96 and 38.65 mm, respectively. Phylogenetic analysis showed that LSN02 and LLGJ04 belonged to Bacillus sp. and existed chemotaxis. In pot experiments, LSN02 and LLGJ04 significantly enhanced the resistance of tobacco plants against bacterial wilt. The rate of tobacco plants infected by tobacco bacteria wilt significantly decreased under LSN02 treatment at a medium (1×10⁸ cfu/mL) or high (2×10⁸ cfu/mL) concentration.