【作者】 何礼远； 冯洁；

【Author】 HE Li-yuan , FENG Jie (State Key Laboratory for Bblogy of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100094)

【机构】 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；

【摘要】 应用有关现代分子生物学新技术结合经典植物病理学、病原细菌学实验方法,获得了青枯病原细菌的三种致病相关胞外蛋白编码基因的克隆和亚克隆,进行了该基因所含核苷酸序列测定和同源性分析,同时还获得了控制青枯菌胞外蛋白输出细胞外膜的基因克隆和亚克隆,也进行了其核苷酸序列测定和同源性分析;表明它们与几种动植物病原细菌的分泌基因具较高同源性。成功地获得了马铃薯抗菌蛋白API编码基因的克隆,测定了核苷酸全序列,表明其与磷酸脂酶等抗逆性酶类具有较高同源性,已得到了该基因在大肠杆菌中的表达,成功地构建了该基因的植物表达载体,完成了该基因的外植体转化,首次获得了API编码基因的分子表达和抗青枯病表达的马铃薯植株。初步证明API可与数种马铃薯蛋白相互作用,其中包括叶绿体中的铁硫蛋白和果糖双磷酸醛羧酶。更多还原

【Abstract】 By applying novel molecular bio-techniques combined with classical methods in plant pathology and bacteriology, 3 pathogenesis related proteins from Ralstonia so- lanacearum were identified, the ecoded genes were cloned or subcloned and the nucleotide sequences were clarified.Meanwhile, the gene for export of extracellular proteins of this pathogen was also cloned, subcloned and sequenced. Molecular analysis revealed that these genes were homologous with extracellular proteins secreting genes of several animal and plant bacterial pathogens. An anti - microbial protein (API) encoded gene, named ap1 from the BW resistant potato variety MS - 42.3 was successfully cloned and its nucleotide fragment was completely sequenced. Results of molecular analysis indicated that the API was homologous with some plant drought - resistant enzymes, such as acid phosphotase and phosphoesterase C. The fusion protein encoded by ap1 was well expressed in E.coli.The API gene was successfully integrated into potato susceptle cultivars and efficiently expressed.Evaluation testing of transgenic potato plants indicated a significantly increased resistance to bacterial with caused by R . solanacearum .The API was proven to be able to interact with several other proteins in potato, including cytochrome iron-sulfur protein and fructose 1,6bisposphatealdolase in chloroplasts.更多还原