项目名称: 利用蛋白质组学策略分析星星草根盐胁迫应答氧化还原敏感蛋白质
项目编号: No.31270310
项目类型: 面上项目
立项/批准年度: 2013
项目学科: 生物科学
项目作者: 戴绍军
作者单位: 东北林业大学
项目金额: 78万元
中文摘要: 星星草(P.tenuiflora)是具有很强耐盐能力的禾本科牧草。研究其盐胁迫应答机制对于筛选重要耐盐基因、提高作物耐盐性具有重要意义。星星草根中的抗氧化系统与蛋白质氧化还原状态转换在盐胁迫应答过程中起着关键作用。本研究拟利用新发展起来的Cy3/Cy5荧光标记技术、同位素标记的亲和标签(ICAT)技术和半胱氨酸反应串联质量标签(CysTMT)技术3种彼此互补的氧化还原蛋白质组学研究策略,解析星星草根应答不同类型/强度盐胁迫过程中硫醇氧化还原敏感蛋白质的表达丰度与修饰状态,并将重要蛋白质编码基因转入拟南芥中,在盐和过氧化氢胁迫条件下,利用Cy3/Cy5染色、Western杂交、蛋白质凝胶阻滞等技术验证蛋白质应对氧化胁迫的功能。通过与根氧化胁迫应答相关细胞学与生理学特征整合分析,构建硫醇氧化还原敏感蛋白质参与调控的受不同盐信号诱导的盐胁迫应答网络机制,为深入解析植物耐盐机制提供更多证据与线索。
中文关键词: 星星草;盐碱胁迫;分子机制;蛋白质氧化还原;蛋白质组
英文摘要: Puccinellia tenuiflora is a monocotyledonous halophyte species belonging to the Gramineae and has outstanding nutritional value for livestocks. It can thrive in the saline-alkali soil of the Songnen plain in northeastern China. Some physiological and proteomics mechanisms in leaves of P. tenuiflora in response to salt/alkali treatments have been highlighted in our previous proteomics investigation. However, the salt tolerance mechanism in roots is still unclear so far. In the fine-tuned sophisticate molecular networks for salt tolerance, the anti-oxidative mechanism and the thiol redox-regulated modification of proteins in roots play key roles. Further investigation of the thiol redox-sensitive proteins involved in salt response will provide valuable information for screening of salt-tolerant genes and improving crops tolerance. In this study, we plan to investigate the salt-responsive expression patterns (e.g. abundance and redox state) of thiol redox-sensitive proteins in P. tenuiflora roots using high throughput quantitative redox proteomics approaches. Three complementary labeling strategies will be used for analyzing thiol redox-sensitive proteins in roots. They are Cy3/Cy5 labeling method, isotope coded affinity tag (ICAT) method, and cysteine-reactive tandem mass tag (CysTMT) technology. Based on these m
英文关键词: Puccinellia tenuiflora;Alkali salinity stress;Molecular mechanism;Protein redox;Proteomics