The invention relates to a bacillus amyloliquefaciens for inhibiting bird rape plasmodiophora brassicae and separation, application and purposes of active substances thereof. Bacillus amyloliquefaciens HB-26 is preserved in China Center for Type Culture Collection (CCTCC), the preserving number is CCTCCNO: M2011259. A culture method is as follows: inoculating biogas slurry separated bacterial strain in an LB liquid nutrient medium, preserving the biogas slurry separated bacterial strain at 30 DEG C for one night, pouring fermentation liquor at the root of Chinese cabbage and finding the bacterial strains with bacteriostatic activity on plasmodiophora brassicae. The separated active substance is characterized in that the HB-26 is inoculated in the LB liquid nutrient medium, the fermentation liquor is dried by freezing, the frozen strain powder is added with ethyl acetate for extraction, is vibrated, rotated and evaporated, is dissolved by methyl alcohol, and then subjected to HPLC (High Performance Liquid Chromatrography) separation, the components are collected, the mass spectrum identification is conducted, and the component of the active substance can be judged according to the condition of the generation of plasmodiophora brassicae disease of the Chinese cabbage. The molecular weight of the bacteriostatic activity substance BA20 is 1494.0, and the nuclear magnetic resonance hydrogen spectrum shows that the chemical nature of the bacteriostatic activity substance is carbohydrate bacteriocin. The active substance is used for preparing a microbial agent for preventing the plasmodiophora brassicae disease of the Chinese cabbage.