[Objective] To provide biocontrol bacteria to control various plant fungal diseases, this study aimed to explore the antagonistic compounds from Bacillus methylotrophicus against Cercospora zeae-maydisTehon et Daniels, Alternaria alternate and Botrytis cinerea. [Methods] Plate method was used for prescreening, and the cup and saucer method was applied for rescreening. The antagonistic strains were identified by cell morphology and 16S rRNA gene. Antagonistic compounds were analyzed by thin-layer chromatography and their coding genes were amplified by the polymerase chain reaction. Corn field trial of biocontrol was used to assess the control effect of antagonistic strain on three pathogenic microorganisms. [Results] Bacillus methylotrophic B-1841 inhibited A. alternata, C. zeae-maydis and B. cinerea with 65.95%, 71.04% and 46.69%, respectively. The antagonistic compounds were identified as iturin-like lipopeptide. The corn field trial showed that strain B-1841 had significant control effects on diseases infected by C. zeae-maydis, A. alternate and B. cinerea, with relative control effects of 69.89%, 60.25% and 45.21%, respectively. [Conclusion] B. methylotrophic B-1841 has a potential application prospect in the prevention and treatment of fungal diseases of crops.