硅调控特定基因提高水稻修复UV-B伤害的分子机制研究

项目名称： 硅调控特定基因提高水稻修复UV-B伤害的分子机制研究

项目编号： No.31300336

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 生物科学

项目作者： 方长旬

作者单位： 福建农林大学

项目金额： 24万元

中文摘要： 氯氟烃等有害气体的大量排放严重破坏了大气臭氧层，由此导致地表紫外线UV-B(280-315nm)辐射增强，使得水稻等作物的DNA受损，生长受抑制，生物量减少，严重影响其生长发育。前期研究表明硅营养能够增强水稻光裂解酶(PL)基因表达进而提高修复UV-B伤害的能力，但水稻中具有4个PL基因，其中哪个基因在此过程中更为高效还未知。据此，本研究拟通过qRT-PCR检测明确高效参与硅提高水稻修复UV-B伤害的PL基因，克隆其启动子，采用固相DNAseI足迹法获得该PL基因启动子的转录因子，并利用EMSA及Western blot技术验证转录因子与启动子的有效互作。在此基础上，应用RNAi及Overexpression技术，验证转录因子及该PL基因对提高水稻修复UV-B伤害的作用，从而揭示硅营养通过特定PL基因提高水稻修复UV-B伤害的分子机制，为利用此技术提高水稻抗UV-B辐射提供理论依据。

中文关键词： 水稻；紫外线B辐射；硅；光修复；转录调控

英文摘要： Depletion of the stratospheric ozone (03) layer due to the emission of chlorofluorocarbons and other trace gases has resulted in increased solar ultraviolet-B (UV-B: 280-315 nm) radiation on the earth's surface, which would increase DNA damage,and reduce aboveground biomass and plant height. These effects of UV-B radiation take serious influence on plant growth and development. The previous studies showed that silicon (Si) contributed to enhance gene expression of photolyase (PL), which can repair UV-B damage on rice plant, however, there are 4 PL genes in rice genome, and which PL gene presents high effciency on this process is still unknown. In order to find the specific PL gene and explore the underlying mechanism,the present research will use qRT-PCR to determinate the differential expression of PL genes, which aims to obtain the specific PL gene with high efficiency on repairing UV-B damage on rice.The promoter of the specific PL gene will be amplified to screen their interacted transcription factors by using Solid-phase DNAse I footprinting method, and then EMSA and western-blot technologies will be used to confirm the positive interactions between gene and transcription factors. Further,both RNAi and overexpression technologies will also be performed to investigate the positive function of the transcripti

英文关键词： Rice；UV-B radiation；Silicon；Photorepair；Transcriptional regulation