短小芽孢杆菌TUBP1抗棉花黄萎病菌活性成分及作用机理研究

项目名称： 短小芽孢杆菌TUBP1抗棉花黄萎病菌活性成分及作用机理研究

项目编号： No.31260013

项目类型： 地区科学基金项目

立项/批准年度： 2013

项目学科： 生物科学

项目作者： 曾红

作者单位： 塔里木大学

项目金额： 48万元

中文摘要： 棉花黄萎病是棉花的"癌症"，利用微生物及其制剂进行棉花黄萎病生物防治是有效途径。本课题以南疆连作15年棉花土壤中分离对棉花黄萎病菌具拮抗作用的短小芽孢杆菌TUBP1菌株为研究对象，通过液体发酵获得发酵产物，并用现代分离技术如HPLC以及1D和2DNMR鉴定技术分离鉴定其发酵液中活性成分；离体活体实验确定活性成分对棉花黄萎病菌抗菌活性；碘化丙啶染色法确定其对棉花黄萎病菌细胞膜的影响；罗丹明染色法测定其作用棉花黄萎病菌后线粒体膜电势、XTT法测定线粒体脱氢酶活性、染色法测定线粒体内活性氧含量；透射电镜观察其作用于棉花黄萎病菌后细胞超微结构的改变，共聚焦显微镜观察DAPI染色后细胞核的形态变化；探讨抗菌活性成分可能的作用机理，为进一步开发防治棉花黄萎病的微生物药物提供理论依据和技术支撑，同时，在保护我国微生物资源、加快微生物资源的有效利用和持续发展、以及促进新疆地方经济的发展方面具有重要意义。

中文关键词： 阿萨尔基亚芽孢杆菌 TUBP1；抗棉花黄萎病菌活性；TUBP1 蛋白；作用机理；

英文摘要： Cotton verticillium wilt is cotton cancers. The biological control, used by microbial agents and microorganism, is the effective way to control cotton verticillium wilt. A antagonistic Bacillus pumilus TUBP1, isolated from 15-years cultured cotton field, is regard as research objectives by our project team. The fermented products will get through the liquid fermentation and it will be subject to isolation and identification research of function components by using mordern seperation methods such as HPLC as well as mordern spectroscopic technology including 1D and 2D NMR. In vitro and in vivo activities of active compounds from Bacillus pumilus TUBP1 against Verticillium dahiae Kleb. Effect of dill active compounds on membrane of Verticillium dahiae Kleb by pyridine iodine staining;We assayed the mitochondrial membrane potential (mtΔψ) using rhodamine 123 and detected the antivities of mitochondrial dehydrogenases by spectrophotometry; We observed the ultrastructure by transmission electron microscopy, studied the nuclear fragmentation by DAPI staining, The mechanism of action of active compunds against Verticillium dahiae Kleb were studied. It is provide the theory basis and the technical support for the further development Bacillus pumilus TUBP1 as microbial medicine to control cotton verticillium wilt. At the

英文关键词： Bacillus axarquiensis TUBP1；Anti-V. dahliae activity；TUBP1 protein；mechanism；