摘要
【目的】利用高速逆流色谱从油松毛虫病原真菌布氏白僵菌CGMCCNo.2382菌株代谢浓缩液的乙酸乙酯粗提物中分离纯化具有杀虫活性的小分子毒素物质。【方法】用乙酸乙酯对白僵菌发酵液萃取,利用高速逆流色谱对粗提物进行分离,其两相溶剂系统为:正己烷-乙酸乙酯-甲醇-水(3.5﹕5﹕3.5﹕5);利用GC/MS对分离物进行检测;并用饲喂、接触和注射法在油松毛虫幼虫上作毒性试验。【结果】从乙酸乙酯的萃取物中分离纯化得到2-香豆满酮、间甲基苯甲酸甲酯和对甲基苯甲酸甲酯3种毒素物质,GC检测其纯度分别为81%、89%和80%;毒性测定结果表明,间甲基苯甲酸甲酯对油松毛虫幼虫的死亡率分别是饲喂法34.44%、接触法35.56%、注射法87.78%,而对甲基苯甲酸甲酯对油松毛虫幼虫的死亡率分别是37.78%、38.89%、91.11%。【结论】间甲基苯甲酸甲酯和对甲基苯甲酸甲酯为布氏白僵菌菌株(CGMCCNo.2382)的代谢毒素。
【Objective】 High-speed counter-current chromatography （HSCCC） was applied to the isolation and purification of small molecule toxins from metabolites of Beauveria brongniartii （CGMCC No.2382）, the pathogenic fungus infecting Dendrolimus tabulaeformis. 【Method】 Ethyl acetate was used to extract toxin components from row extraction of secondary metabolites. The extraction was isolated and purified by HSCCC with two-phase solvent system composed of hexane-ethyl acetate-methanol-water （3.5﹕5﹕3.5﹕5, V/V）, and the compounds were identified by GC/MS. The toxicity bioassay was conducted by breeding, epidermis contact and injection. 【Result】 Three compounds, 2-coumaranone,3-methyl-benzoic acid methyl ester and 4-methyl-benzoic acid methyl ester, were obtained with 81%, 89% and 80% in purity by GC method, respectively, from secondary metabolites. The bioassay showed that the toxin 3-methyl-benzoic acid methyl ester resulted the mortality of 34.44% in breeding, 35.56% in epidermis contact and 87.78% in injection. While the toxin 4-methyl-benzoic acid methyl ester caused the mortalities of 37.78%, 38.89% and 91.11%, respectively. 【Conclusion】 3-methyl-benzoic acid methyl ester and 4-methyl-benzoic acid methyl ester are active toxins of Beauveria brongniartii （CGMCC No.2382） to the host insect.
出处
《中国农业科学》
CAS
CSCD
北大核心
2010年第15期3125-3133,共9页
Scientia Agricultura Sinica
基金
山西省科技攻关项目(2007032009)
关键词
布氏白僵菌
毒素
高速逆流色谱
油松毛虫
Beauveria brongniartii
toxin
high-speed counter-current chromatography
Dendrolimus tabulaeformis
