Abstract: In this study, the inhibitory effects of Bacillus subtilis strain BS-1 and the sterile fermentation broth on Botryosphaeria dothidea and Diaporthe nolilis（the main pathogens of ‘Xuxiang’ kiwifruit soft rot） were investigated, as well as the preservation effects of shelf life. Results showed that the Bacillus subtilis strain BS-1 and the sterile fermentation broth could significantly inhibit the growth in vitro and fruit pathogenicity of B. dothidea and D. nobilis. The FJY showed good antibacterial ability in the temperature range of 37～100 ℃, ultraviolet irradiation time range of 0.5～2 h, and pH range of 3～5 and 8～10. Besides, BS-1 and FJY treatments could significantly delay the softening of fruit, improve the fragility and tightness, retard the rise of soluble solids content and the decrease of titratable acids content, and reduce the weight loss rate and rot rate, thereby improving the postharvest qualities of kiwifruit. The FJY treatment presented superior preservation effects than BS-1. This study would provide a theoretical basis for the biological control of kiwifruit soft rot and a new idea for preservation of postharvest kiwifruit.