Bacillus subtilis S1-0210 was selected as a biological agent against Botrytis cinerea in strawberry. The isolate inhibited mycelial growth of B. cinerea in vitro tests. A wettable powder formulation of B. subtilis S1-0210 significantly reduced infection rates with lower than 5%, compared with higher than 70% of infection rates in untreated control. The formulation showed 85 to 89% control efficacies of gray mold incidences on fruits of strawberry in pots. Pre-treatment of the agent was more effective in controlling gray mold on fruits and leaves than post-treatment at the early stage of disease development. The formulation also showed 70% control efficacy of gray mold incidence on fruits of strawberry in a field trial. The results indicate that B. subtilis S1- 0210 in the wettable powder formulation may be a potential biocontrol agent to control gray mold on strawberry.