This study was carried out to clarify the effects of antagonistic microorganism, Bacillus sp. JC181 isolated from the greenhouse soil grown cucumber plants on the growth inhibition of plant pathogen, fusarium wilt (Fusarium oxysporum) occurred in cucumber plants in greenhouse. Antagonistic bacterial strains were isolated and were investigated into the antifungal activity of the antagonistic microorganism against fusarium wilt. Screened fourteen bacterial strains which strongly inhibited F. oxysporum were isolated from thc greenhouse soil grown cucumber plants, and the best antagonistic bacterial strain designated as JC181, was finally selected. Antagonistic bacterial strain JC181 was identified to be the genus Bacillus sp. based on the morphological and biochemical characterization. Bacillus sp. JC181 showed 58.2% of antifungal activity against the plant pathogen growth of F. oxysporum. By the bacterialization of culture broth and heated filtrates of culture broth, Bacterial strain, Bacillus sp. JC181. showed 91.2% and 260% of antifungal activity against F. oxysporum, respectivrly.