棉花黄萎病是危害棉花的严重病害，我国棉花黄萎病的病原菌为大丽轮枝菌（Verticillium dahliae）。从拮抗菌中提取抑菌物质是防治黄萎病的生物防治手段之一。 从新疆棉田土壤中分离出来的枯草芽孢杆菌J-15对大丽轮枝菌具有拮抗作用。本实验采用枯草芽孢杆菌J-15作为实验材料，探究提取活性物质的分离纯化过程，为活性物质的鉴定奠定基础。 J-15用牛肉膏蛋白胨培养基发酵，当培养基pH=7时，能产生最好的抑菌效果。将J-15以1:100的比例扩大培养，12500r/min离心15min去除菌体，获得无菌发酵液。调节无菌发酵液pH为2，过夜静置后13000r/min离心20min，弃去上清液，收集沉淀。使用不同的有机溶剂抽提沉淀中的活性物质，显示80%丙酮的效果最好。 经旋转蒸发除去溶液中的丙酮后，加用水溶解。在水溶液中加入一倍氯仿剧烈震荡，12000r/min离心10min，取上清，加入一倍水饱和的正丁醇，离心后得到溶于正丁醇的活性物质粗提液。正丁醇粗提液静置后分为上清和沉淀。将正丁醇溶液上清和沉淀进行硅胶柱层析和薄层层析（Thin layer chromatography ，TLC）分析。结果表明在正丁醇上清液中，有两种环脂肽存在；在正丁醇的沉淀中，有一种环脂肽存在。

Cotton Verticillium wilt is a serious disease for cotton. The pathogen of it in China is Verticillium dahliae. Extracting bacteriostatic substances from antagonistic bacteria is one of the biological control methods for controlling Verticillium wilt. Bacillus subtilis J-15 isolated from cotton soil in Xinjiang has an antagonistic effect on Verticillium dahliae. In this experiment, Bacillus subtilis J-15 was used as experimental material to explore the separation and purification process of extracting active substances, which laid a foundation for the identification of active substances. J-15 is fermented with beef extract peptone medium, which produces the best bacteriostatic effect when the medium pH=7. The J-15 was expanded in a ratio of 1:100, and the cells were removed by centrifugation at 12500 r/min for 15 min to obtain a sterile fermentation broth. The pH of the sterile fermentation broth was adjusted to 2, centrifuged at 13000 r/min for 20 min after standing overnight. Then the supernatant was discarded to collect the precipitate. Extraction of the active material in the precipitate using different organic solvent, the study showed that 80% acetone worked best. After rotary evaporation to remove acetone from the solution, it was dissolved in water. The aqueous solution was vortexed by adding chloroform, centrifuged at 12000 r/min for 10 min, and the supernatant was taken. One-times of water-saturated n-butanol was added to the supernatant, and after centrifugation, a crude extract of active substance dissolved in n-butanol was obtained. The crude n-butanol extract was separated into supernatant and precipitate after standing. The n-butanol solution supernatant and precipitate were subjected to silica gel column chromatography and thin layer chromatography (TLC) analysis. The resμLts showed that there were two cyclolipopeptides present in the n-butanol supernatant; in the precipitation of n-butanol, a cyclolipopeptide was present.