非标记定量蛋白质组学在烟草青枯菌致病性与研究中应用_宋浩.pdfVIP

分子植物育种，2010 年，第8 卷，第3 期，第595-604 页 Molecular Plant Breeding, 2010, Vol.8, No.3, 595-604 技术改进 Upgraded Technology 非标记定量蛋白质组学在烟草青枯菌致病性研究中的应用 1* 2* 2 2 1 2** 宋浩 沙伟 丁伟 刘翀 周燕 陈薇 1 湖南中烟工业有限责任公司, 长沙, 410014; 2 上海中科新生命生物科技有限公司, 上海, 200233 * 同等贡献作者 ** 通讯作者, wchen@ 摘 要 烟草青枯病是由茄科劳尔氏菌(Ralstonia solanacearum) 引起的一种毁灭性的烟草种植灾害，可造成 烟草的巨大减产。本实验室应用蛋白质组学中的Shotgun 技术、非标记质谱定量方法和MA plot 等对致病和 非致病青枯菌蛋白质进行了生物信息学分析，从蛋白质组水平研究烟草青枯菌的蛋白质结构和功能，为了 解其致病机理和防治提供新的方向。本文采用Shotgun 技术在非致病性和致病性菌株中分别找到 1 628 和 1 346 个蛋白质，其中287 个为非致病性菌株中特有，15 个为致病性菌株中特有。对于两种菌株共有的1341 个蛋白质用MA plot 方法分析得到了163 个显著性量变差异，并采用GO 富集分析，显示它们主要定位于细 胞外(intracellular part)或与糖代谢相关，为进一步揭示青枯菌侵染烟草的生物学机制提供了有力的支持。 关键词 非标记蛋白质组学, 烟草青枯菌, 致病性 The Application of Laber-free Quantification Proteomics in Tobacco Ralsto－ nia solanacearum Pathogenicity Research Song Hao 1* Sha Wei 2* Ding Wei 2 Liu Chong 2 Zhou Yan 1 Chen Wei 2** 1 China Tobacco Hunan Industry Co. Ltd., Changsha, 410014; 2 Shanghai Applied Protein Techology Co. Ltd., Shanghai, 200233 * The authors who contribute equally ** Corresponding author, wchen@ DOI: 10.3969/mpb.008.000595 Abstract Ralstonia solanacearum (synonym pseudomonas solanacearum) is a widely spread soli-borne plant pathogen which can cause “bacterial wilt”, a major bacterial plant diseases that greatly impact the production of to- bacco. In this experiment, we discuss the structure and function of Ralstonia solanacearum on protein level by em- ploying Shotgun method, label-free mass spectrometry relative quantitative strategy and MA-Plot, that make a new basis for its pathogenicity and bio-protection research. As our result, 1 628 and 1 346 proteins were identified by the shotgun method in avirulent and virulent strains, respectively. 287 proteins are unique in avirulent stains and 15 proteins in virulent strains. For the common 1 341 proteins in both strains, we finally identified 163 differential expressed proteins through MA-Pl