大豆生育期基因E1的作用机理研究

项目名称： 大豆生育期基因E1的作用机理研究

项目编号： No.31271742

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 农业科学

项目作者： 夏正俊

作者单位： 中国科学院东北地理与农业生态研究所

项目金额： 86万元

中文摘要： 大豆生育期基因E1是控制开花期与成熟期的主效基因，且与光周期反应密切相关。申请者已成功克隆出位于着丝点附近的E1基因，初步明确其为豆科植物特有的转录因子。本项目将集中解析尚未开展研究的E1基因的作用机理,在分子水平上明确E1与成花素GmFTs间的互作关系。基于E1含有核定位信号，首先明确E1的突变体对核定位的影响，进而分析其为转录激活或转录抑制作用。应用染色质免疫沉淀-高通量测序技术，在全基因组范围内解析E1基因的靶体位点；同时利用化学诱导型启动子XVE 系统来鉴定E1直接调控的下游基因。以GmFTs基因为重点,应用ChIP-qPCR、酵母单杂交、凝胶阻滞等技术来检验E1与靶体位点间互作关系。利用苜蓿突变体库存中E1及靶体位点的同源序列突变体进一步完成功能验证。在分子水平上明确E1调控下游基因及GmFTs的机理，对于揭示大豆开花基因的调控网络及开展生育期的分子育种具有重要理论与现实意义。

中文关键词： 大豆；开花期；E1；转录因子；作用机理

英文摘要： The maturity locus E1 is the major gene underlying the flowering time and maturity in soybean, also involved in the photoperiodic response. Using positional cloning approach, the applicant and his colleagues have successfully identified molecular identity for the maturity E1 locus, which resides in the pericentromeric region and is difficult to identify. Previous studies revealed that this gene is a novel type of legume-specific transcription factor (TF). This research project will focus on the functional mechanism of the E1 gene with an emphasis on how E1 regulates GmFTs, the components of florigens, directly or indirectly. Given E1 gene has a nuclear localization signal (NLS) and DNA binding sites, we will first investigate whether the mutation in the E1 gene has any impact on subcellular distribution of the E1 protein, then to detect whether the E1 gene carries transcriptional activation or repression activity. Importantly, we use the chromatin immunoprecipitation-sequecing (ChIP-seq) to survey the genome-wide binding sites of the E1 protein. Also we use the XVE inducible expression system to investigate the direct target gene(s) of the E1 protein. The functional relationship between E1 and GmFTs (GmFT2a/GmFT5a) as well as other newly identified target sites will be validated by chromatin immunoprecipitation

英文关键词： Soybean；Flowering time；E1；Functional mechanism；Transcription factor