Abstract: Biocontrol bacteria SF-16 was a selected bacteria strain,research showed that its inhibitory effect on multiple plants.In order to improve biocontrol bacteria SF-16 spore formation rate and quantity,provide theoretical basis for factory production of biological agent,use flask fermentation with a survey on main influencing factors of spore formation.Through single factor experiment and orthogonal experiment,the optimal shaking flask fermentation condition was determined as follow：media composed of 0.5% lactose,2.0% soybean meal,0.30% MnSO4·5H2O.Under this optimum condition,the germination rate achieves 98.3 percentages,Biomass achieves 2×109/mL.