水稻盐敏感突变基因rss2的精细定位与克隆

A rice mutant exhibiting high salt sensitivity in seed germination, seedling, and booting stages has been isolated and identified, and named rss2（rice salt sensitive 2）. The rss2 locus was mapped primarily in our previous work. In this project, we will define and clone this putative gene by map-based cloning methods. The gene function in salt tolerance will be further confirmed by transgenic complementation, RNAi, and overexpression of RSS2. Using GUS and real-time PCR, RSS2 expression in different tissues will be determined both qualitatively and quantitatively with and without salt treatment. RSS2 localization in cell will be investigated by tagging green fluorescence protein (GFP). Furthermore, the gene will be primarily characterized using bioinformatic and molecular approaches. The aim of this project is to ascertain the molecular mechanisms of the RSS2 gene in regulation of intracellular Na+ levels and salt tolerance in rice.

我们从EMS 诱导的水稻突变群体中，筛选、鉴定到了一个在种子萌发、幼苗、孕穗等时期均表现对盐胁迫敏感的突变体rss2（rice salt sensitive 2），并进行了初步的基因定位。本项目拟将RSS2基因定位到更小的染色体区段内，并通过图位克隆方法将其分离。利用转基因功能互补验证、RNA干扰、过表达等手段，明确RSS2基因与水稻耐盐性的关系。采用GUS-RSS2、定量PCR等技术，定性、定量研究RSS2在组织中的表达，及其与耐盐性的关系。利用GFP-RSS2,对其亚细胞定位进行分析。进一步利用生物信息学和分子生物学手段，初步鉴定RSS2的生物学特性及功能，为揭示该基因在调控水稻细胞内Na+水平及耐盐的分子机制奠定基础。