盐胁迫下黄瓜SAMs蛋白的表达定位与功能分析

S-腺苷甲硫氨酸合酶(SAMs)是植物蛋白质组学研究中发现的一个参与众多胁迫响应的重要蛋白。前人及我们前期研究表明，黄瓜SAMs蛋白具有强烈的盐诱导特性，但其具体的功能仍不清楚。本研究准确控制根际生长环境，通过正常营养液和盐胁迫处理黄瓜幼苗，运用荧光定量PCR和蛋白免疫印迹技术分析盐胁迫下黄瓜SAMs基因和蛋白的表达特性；利用荧光免疫组化、原生质体转化和免疫胶体金技术分析SAMs蛋白在组织中和细胞内精确定位；采用免疫共沉淀技术、结合质谱技术分离鉴定与SAMs相互作用的蛋白，并进行互作网络关联分析，探讨黄瓜SAMs在响应盐胁迫中的调控作用；通过构建过表达和反义载体，获得转基因株系，对SAMs的耐盐功能进行验证，进而揭示SAMs蛋白在植物耐盐性中的作用机制。研究结果可丰富植物逆境生理理论，对设施作物抗逆促长优质高效栽培和抗性品种选育均具有重要的实际意义。

S-adenosylmethionine synthase (SAMs) is an important protein that involved in plant stress responses by proteomics studies. Early reports and our previous study showed that SAMs was induced significantly by salt stress, but its detailed function on salt tolerance is unclear. In this study, in order to control the growing environment in the rhizosphere accurately, we will use the nutrient solution cultivation with the normal nutrient solution and salt stress treatment on cucumber seedlings. We will analyze the expression of SAMs gene and protein characteristics by fluorescence quantitative PCR and Western blot technology. Intracellular and tissue of SAMs protein localization will be analyzed by fluorescent immunohistochemistry and protoplast transformation and colloidal gold technical, separation and identification of interacting proteins with the SAMs by using the co-immunoprecipitation and mass spectrometry. Meanwhile we will analyze the associated interaction network, in order to explore the regulation of SAMs on salt stress responses. We will obtain transgenic lines to verify the functions of SAMs under salt stress by building over-expression and anti-sense vector, and further to reveal the mechanisms of SAMs protein on salt tolerance. Our results are beneficial for enriching plant stress physiology theory, and have important practical significance on cucumber cultivation of quality and efficient and resistance breeding.