Two biocontrol bacterial strains of BRF-1 and BRF-2 were isolated from soybean rhizosphere with broadly antifungal activity.Both strains can growth under high temperature, and are Gram positive and forming endospores in the cell.BRF-1 has no flagellum, and BRF-2 has flagella and grew around cell.The physiological characteristics of BRF-1 and BRF-2 are different.Identification by using the methods of Biolog and 16S rDNA sequence analysis showed that BRF-1 is Paenibacillus polymyxa and BRF-2 is Bacillus subtilis.BRF-1 has highest cells quantity and antagonistic activities after it have been cultured for 24 hours, and the antagonistic substance was identified as peptide with the characteristics of thermostable, resistance to acid and proteinase-K.The peptide was separated by ammonium sulfate and purified by Sephadex G-50, and acquired short peptide with molecular weight about 35.4 kDa, which has antifungal activity against Rhizoctonia solani.Culture medium of PSS, 2P and PD are beneficial to BRF-1 to product peptide than other medium.For BRF-2, It was found that after 72 hours cultivation, the antagonistic activities and OD600 of BRF-2 reached the highest level, and the antagonistic substance is confirmed as non-protein and hydrosoluble antibiotic with the characteristics of partial sensitive to high temperature, strong acid and alkali.BRF-2 doesnt request special medium for antibiotic produce.Pot and field experiments showed that the suspension or metabolic solution of both strains not only effectively control Fusarium wilt disease of cucumber and tomato and Fusarium root rot disease of soybean, and also significantly promote those seedling growths.In addition, the influences of inoculating two bacterial strains on bacterial and fungal community structures in the rhizosphere soils of cucumber and tomato were estimated by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) method.Results showed that the inoculation with two strains had little effects on the structure of bacterial community of both vegetables, but it had changed the fungal community of tomato and no significant changed the fungal community of cucumber.