Bacillus subtilis RB14-C, which is a streptomycin resistant mutant of B. subtilis RB14 isolated from compost, produces an antifungal peptide, iturin A, and was evaluated for its suppressive ability against damping-off of tomato seedlings and Phomopsis root rot of cucumber. In damping-off disease of tomato seedlings caused by Rhizoctonia solani, RB14-C cell suspension treatment did not suppress the disease occurrence whereas germinated seed treatment of the RB14-C cells reduced the occurrence of the damping-off. In both treatments, iturin A could not be detected from the treated soil. In the Phomopsis root rot of cucumber, root immersion treatment of cucumber seedlings with RB14-C cell suspension at the time of transplanting effectively suppressed the root rot, resulting in growth recovery 50 days after the treatment even though the initial growth was retarded due to the Phomopsis infection. These results suggest that germinated seed treatment of RB14-C cells and root immersion treatment with RB14-C cell suspension can be applied as promising biological control practices of the damping-off of tomato seedlings and the cucumber Phomopsis root rot, respectively.