植物青枯菌aac 基因克隆及猝灭群体感应信号功能的研究 - 中国农业科学.pdf

中国农业科学 2008,41(9):2651-2656 Scientia Agricultura Sinica doi: 10.3864/j.issn.0578-1752.2008.09.011 植物青枯菌 aac 基因克隆及猝灭群体感应信号功能的研究 张 争，徐 进，许景升，何礼远，冯 洁 （中国农业科学院植物保护研究所植物病虫害生物学国家重点实验室，北京 100094 ） 摘要：【目的】研究植物青枯菌（Ralstonia solanacearum ）aac 基因编码的蛋白是否具有降解细菌群体感应 信号分子的功能。【方法】PCR扩增获得青枯菌GMI 1000菌株的aac 基因，将 aac 基因克隆到pET-5a 原核表达载 体上，在大肠杆菌中表达出 AAC 融合蛋白，将 AAC 融合蛋白与胡萝卜软腐欧氏杆菌（Erwinia carotovora sp.carotovora ）混合后，共接种于马铃薯块茎，研究细菌致病力的变化。【结果】克隆了全长的aac 基因，完成 了aac 基因原核表达载体的构建，研究了 AAC 融合蛋白对细菌致病力的影响。【结论】青枯菌aac 基因编码的蛋 白具有减弱胡萝卜软腐欧氏致病力的功能，为开发新的控害策略提供了依据。 关键词：植物青枯菌；aac 基因；克隆；群体感应信号；猝灭 Cloning of aac Gene of Ralstonia solanacearum and Function Research on Quenching Quorum-Sensing Signal ZHANG Zheng, XU Jin, XU Jing-sheng, HE Li-yuan, FENG Jie (State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100094) Abstract: 【Objective 】To determine whether protein product encoded by aac gene of Ralstonia solanacearum possesses a function of quenching bacteria quorum-sensing signal. 【Method 】 aac gene was cloned by PCR amplification, and constructed into pET-5a expressing vector. AAC fusion protein was obtained in E. coli system. To determine the effect of AAC fusion protein on bacterial pathogenicity, the mixture of Erwinia carotovora sp. carotovora and AAC fusion protein were inoculated into potato tubers. 【Result 】 The whole aac gene was cloned, and constructed into expressing vector. The function of AAC fusion protein was analyzed. 【Conclusion 】 The result showed that AAC fusion protein possesses capacity of decreasing pathogenicity of Erwinia carotovora sp. carotovora, which provides scientific evidence for exp