中文摘要
根系是植物吸收水分的主要器官,根系基因的克隆与分析是揭示根系发育和抗旱机制的主要途径。我们图位克隆了控制旱稻根粗和根长的QTL qBRT9a,遗传转化和关联分析证明了目的基因OsbHLH120控制根系发育。基于前期基础,本项目拟利用已获得的qBRT9a导入系、超表达和RNAi转基因材料等,采用根系X-光扫描成像、体视镜和超薄树脂切片观察根系构型及根尖各区域细胞数目和长度;通过亚细胞定位、GUS染色、qRT-PCR、ChIP-Seq、表达谱芯片、酵母双杂交与GST pull-down实验,分析OsbHLH120表达部位、对土壤干旱和激素的响应、调控的下游基因及互作蛋白,进而探明OsbHLH120参与根系发育的细胞学机制和分子调控网络,为解析作物根系发育、抗旱机制和培育节水抗旱稻奠定基础。
英文摘要
Roots are the principal plant organs for nutrient and water uptake. A strong root system would enable a plant to extract more water and nutrients from the soil, which is important for good plant growth and increased yield under soil-related stresses. Thus, it is necessary to determine the main genetic determinants of the strong root system and to transfer these genes into lowland rice to improve their drought resistance. A novel QTL, qBRT9a, for root thickness and root length was cloned by our research group. A basic helix-loop-helix (bHLH) transcription factor, LOC_Os09g28210 (named as OsbHLH120) identified as a candidate gene regulates root growth confirmed by genetic transformation and correlation analysis. In this research, using over-expression and RNAi transgenic material, and qBRT9a introgression line, root architecture and the number and the length of cells in different regions of root tip will be investigated through root X-optical scanning imaging, stereoscope, and resin sections. The expression pattern, downstream genes, and protein interaction of OsbHLH120 will be researched using subcellular localization, GUS staining, qRT-PCR, ChIP-Seq, expression microarray, and GST pull-down. Then, cytological evidence and molecular pathway of that qRT9 promotes root growth and the value of qBRT9a in field breeding will be discussed. These results will provide a starting point for functional characterisation of the mechanism of root growth, drought resistance and molecular-assisted selection for improving drought resistance.
