白菜CRFs基因在雌蕊发育中的作用及分子调控机制研究

花分生组织精准的维持和终止是植物雌蕊组织分化和形态建成的关键环节，雌蕊发育在高等植物个体发育和世代交替过程中占有重要地位，对农作物的产量和品质有着根本性的影响。目前我们对CRFs在植物雌蕊发育中的作用及分子机理还知之甚少。本项目拟以白菜‘Bajh97-01B’为材料，在分析BrCRFs的时空表达特征及其对不同细胞分裂素的响应特征的基础上，应用amiRNA方法分析BrCRF1和BrCRF19的生物学功能。同时，采用酵母双杂与BiFC以及GST pull-down与ChIP-seq等技术研究BrCRFs与植物激素信号传导途径相关转录因子间的表达调控关系，阐明BrCRFs在花分生组织的维持和终止过程中的作用。随后，分析miRNA和组蛋白甲基化与BrCRFs间的作用关系，揭示BrCRFs通过转录反馈与表观遗传等途径来调控白菜雌蕊发育的分子机制。研究结果将为实现蔬菜的高效繁育提供研究依据。

Result indicates that the precise maintaining and termination of floral meristem is the critical point for gynoecia tissues differentiation and morphogenesis of gynoecium in plant. Gynoecium development plays an important role in ontogenesis and alternation of generations in higher plant, and has an important effect on crop yield and quality. In this study, the temporal and spatial expression characteristics of CRFs gene and their response to different CK will be studied by qPCR and in situ hybridization (ISH) in Chinese cabbage (cv. Bajh97-01B). Subsequently, the artificial microRNA (amiRNA) method will be employed to analyze the biological functions of BrCRF1 and BrCRF19 from Chinese cabbage, and whether there is functional redundancy will be illuminated. Meanwhile, Yeast two-hybrid, BiFC, GST pull-down and ChIP will be performed to study the regulation and crosstalk between CRFs and transcriptional factors which involved in phytohormone signaling pathway. The function of CRFs will be clarified on the process of the precise maintaining and termination of floral meristem. At last, we aim to analyze the relationship which mediated by CRFs between the miRNA and epigenetic patterns and the termination of floral meristem. Results will be helpful to elucidate the molecular mechanism of CRFs regulation of the pistil development through the transcription feedback and epigenetic regulation in Chinese cabbage. In conclusion, these results of this study will provide insight into the molecular mechanism of sexual reproduction in economic crops. In addition, it will also supply the novel theoretical basics to regulate the fertility and realize the high yield and good quality in seed production in Brassica crops.