光周期介导的开花时间天然变异空心菜种质的分子解析

申请者对发现的短日条件下开花时间存在明显差异的两份空心菜天然种质进行了深入的研究积累。通过图位克隆策略，获得了光周期介导的植物开花时间调控新基因IaSDF1的序列特征。本项目将深入研究IaSDF1调控开花时间的分子机制。将应用RNA聚合酶II（Pol II）抗体结合染色质免疫共沉淀（ChIP）技术分析IaSDF1位点和IaFT2/IaFT3的转录；H3K27me3的抗体结合ChIP技术分析IaFT2/IaFT3的表观调控。通过蛋白标签技术纯化蛋白并结合液相色谱-串联质谱技术（LC-MS/MS）和双分子荧光互补技术（BiFC）鉴定蛋白IaSDF1的互作蛋白。IaSDF1涉及的植物开花时间调控分子机制的深入解析，将拓宽光周期介导的植物开花时间分子机理的认识，并为旋花科植物遗传育种提供重要基因。

Using two natural germplasm of water spinach showing different flowering time under short-day conditions, the new photoperiod-mediated flowering time regulation gene IaSDF1 was obtained by map-based cloning strategy and gene functional analysis. This project will further study the molecular mechanism of IaSDF1 regulating flowering time. Loci IaSDF1 and IaFT2/IaFT3 will be analyzed by RNA polymerase II (Pol II) antibody combined with chromatin immunoprecipitation (ChIP) technique, and the epigenetic regulation of IaFT2/IaFT3 will be analyzed by H3K27me3 antibody combined with ChIP technique. Proteins will be purified by protein labeling technology, then proteins interacting with IaSDFl1 will be identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and bimolecular fluorescence complementation (BiFC). The molecular mechanism of IaSDF1 regulation of flowering time will broaden the understanding of flowering time mediated by photoperiod and provide important genes for the genetic breeding of Convolvulaceae plants.