TIFY 转录因子参与水稻耐寒过程的机理研究

项目名称： TIFY 转录因子参与水稻耐寒过程的机理研究

项目编号： No.31500219

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 生物科学

项目作者： 黄小贞

作者单位： 贵州大学

项目金额： 20万元

中文摘要： 环境低温会严重影响水稻的产量和品质。研究水稻响应低温胁迫的分子机制，对于全面了解水稻的抗寒基础及培育抗寒新品种具有重要意义。为了克隆更多的水稻耐寒基因，我们选择了贵州当地一种耐寒的粳稻品种“公告鬼”(Ggg)作为研究对象，构建了Ggg低温胁迫响应SSH文库，从文库中筛选克隆了一个TIFY类型的转录因子，命名为GCT1 (Gui Chilling- Tolerance 1)。研究发现该基因在粳稻Ggg中受到低温诱导表达，并且GCT1的蛋白序列在粳稻Ggg和籼稻93-11之间存在着一个氨基酸差异。本项目拟通过获得过量表达GCT1和低量表达GCT1的转基因植株，以各项耐寒生理指标和相关基因表达为依据，确定转基因植株的耐寒性，以此对GCT1的功能进行验证；并利用生理生化手段和转录组测序分析技术，探讨GCT1可能的调控水稻低温应答的作用机制，为水稻的抗寒育种提供一定的理论基础。

中文关键词： 水稻；低温；GCT1；TIFY；转录因子；转基因

英文摘要： Low temperature is one of the major environmental factors that influence rice (Oryza sativa L.) growth, development and production. Rice seedlings are particularly sensitive to chilling in early Spring, improvement of chilling tolerance in rice may significantly increase rice production. To identify the genes involved in the increased chilling tolerance found in cultivars from growth regions with low yearly-temperatures, we chose one kind of strong cold-resistance rice, named Gonggaogui (O. sativa ssp. japonica, refered as Ggg) as materials from GuiZhou province. In our previous study, we constructed the cold-resistant gene library through Suppression Subtractive Hybridization (SSH). From the SSH library, we isolated a GCT1 (Gui Chilling-Tolerance 1) gene which maybe involve in the pathway of cold resistance of Ggg. Expression of OsGCT1 was up-regulated by cold temperature, and genomic DNA sequence comparisons between the Ggg and 93-11 (O.sativa ssp. indica) showed that one single-nucleotide mutation results in a change in an encoded amino acid. Here, in order to explore the function of OsGCT1, we will construct GCT1 -overexpression (OE) and RNA interference (Ri) transgenic rice lines in Ggg and 93-11, and investigate their chilling tolerance. Meanwhile, we will analyze the effects of GCT1 on rice yield and quality, to evaluate the potential value of this gene in proving rice chilling-tolerance. In addition, we will check the expression pattern of GCT1 and its protein localization under normal and cold conditions. By means of biochemical approach and transcriptome profiling analyses, we will perform some functional analysis and detect the interaction proteins of GCT1. At last, we try to investigate the role of GCT1 in rice adaptation to chilling temperature. Our studies may reveal a novel pathway that controls cold adaptation in rice and will help to broaden the possibilities for genetically engineering cold-tolerant rice cultivars.

英文关键词： rice (Oryza sativa L.);low temperature;Gui Chilling-Tolerance 1 ;TIFY transcription factors;transgene