The invention discloses bacillus subtilis, a biological prevention and control preparation thereof and an application of the biological prevention and control preparation, and belongs to the field of biological plant disease and pest prevention and control. The stain is bacillus subtilis XLBS-01, and has the preservation number of CGMCC No. 6692. Tests show that the bacillus subtilis disclosed by the invention has the characteristics of high growing speed, large sporulation quantity, wide action spectrum, strong stress resistance, capability of realizing rapid and large-quantity colonization at plant roots, and the like, and therefore, the bacillus subtilis has good application prospects. The biological prevention and control preparation prepared from the bacillus subtilis can not only effectively prevent and control plant soil-borne diseases, but also can effectively increase the crop output, thereby being a biological prevention and control preparation with vast application prospects. The microbial preparation can be used as a biopesticide or a biofertilizer for preventing and controlling various soil-borne plant diseases and also preventing and controlling plant leaf diseases, such as hot pepper anthracnose, cucumber target spot, grape downy mildew, rose powdery mildew and the like.