Abstract: This study aims to screen and isolate Bacillus strains with strong inhibitory effect on coniella diplodiella, and to detect the antifungal activity and identify the stain. Bacillus strains were isolated from soil by dilution-plate method. The Bacillus strain GSBM05 with better antimicrobial activity was studied by the plate confrontation method and mycelial growth rate method. The results showed that forty Bacillus strains were isolated from soil samples. One strain, GSBM05, with potent antimicrobial activity was obtained after primary and secondary screening. The inhibition rates of the 20-fold-diluted aseptic fermentation broth of GSBM05 on spore germination and mycelial growth were both above 90%, and control efficacies of different components of the fermentation broth of GSBM05 on detached grape fruits and leaves were over 80%. The results showed that GSBM05 had obvious antifungal activity against eleven pathogenic fungi, including coniella diplodiella. Based on the morphological observation, physiological and biochemical characteristics, and the sequence analysis of 16S rDNA, strain GSBM05 was confirm to be a kind of Bacillus methylotrophicus. The experimental results demonstrated that Bacillus methylotrophicus strain GSBM05, with its excellent antifungal activity and broad antifungal spectrum, can provide new resources for the biological control of grape white rot.