茄青枯拉尔氏菌Rs-T02编码精氨酸脱亚胺酶基因的功能分析

项目名称： 茄青枯拉尔氏菌Rs-T02编码精氨酸脱亚胺酶基因的功能分析

项目编号： No.31460459

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 黎起秦

作者单位： 广西大学

项目金额： 50万元

中文摘要： 由Ralstonia solanacearum引起的植物青枯病是一种难以防治的土传病害，虽然植物抑菌物质对研究现代农药具有巨大启发价值，但筛选到的高效抑菌物质种类少且研究不够深入。我们的前期工作已证实3,4,5-三羟基苯甲酸甲酯（MG）具有开发为杀菌剂的潜力，青枯病菌受MG作用后产生的精氨酸脱亚胺酶（ADI）是影响MG抑制病菌的关键酶之一。本项目拟在获得病菌编码ADI的基因（arcA）序列基础上，构建重组表达载体，并转入大肠杆菌中表达，利用基因工程手段进行arcA基因缺失突变体的构建以及功能互补，分析arcA基因缺失突变体和功能互补菌株的表型，并对MG作用下的arcA基因缺失突变体蛋白质组学进行研究。研究结果将初步明确茄青枯拉尔氏菌MG-Rs-T02 新增蛋白精氨酸脱亚胺酶的功能，进一步揭示MG对R. solanacearum的抑菌机制，为植物青枯病新型高效特异杀菌剂的开发提供基础。

中文关键词： 茄青枯拉尔氏菌；精氨酸脱亚胺酶；arcA基因；功能分析；3；4；5-三羟基苯甲酸

英文摘要： Plant bacterial wilt, caused by Ralstonia solanacearum, is one of the most devastating soil-borne diseases distributed throughout the world. Plant antibacterial substances provide important inspirations to study modern pesticides. In the screening of antibacterial substances using R. solanacearum as a target bacterium, we found that methyl gallate (MG) from Toxicodendron sylvestre could significantly inhibit the pathogen growth in vitro and in planta and had great potential as a bactericide.Arginine deiminase(ADI）was a extra protein of R. solanacearum upon MG treatment . It was possibly deduced that ADI was closely related to the antibacterial mechanisms of MG against R. solanacearum. In this project, a recombinant ADI (rADI) will be cloned by PCR and be expressed in Eseheriehia eoli. A arcA deletion mutant ΔarcA will be constructed by homologue recombination technology. The function of arcA gene encoding ADI in R. solanacearum will be elucidated by analyzing the phenotype of △arcA and complemented strain and studing the proteomics of arcA deletion mutant ΔarcA. Moreover, the studies can hopefully delineate the pharmacological action site of MG and to provide theoretical basis for developing novel and efficient bacteriacide.

英文关键词： Ralstonia solanacearum;arginine deminase;arcA gene;functional analysis;methyl gallate