MADS-RIN下游基因的鉴定及功能分析

果实是植物特有的发育器官，在种子成熟和传播过程中发挥重要作用。成熟衰老是果实发育的重要阶段，也是果实品质形成和保持的关键时期，研究果实成熟衰老的调控机制具有重要的理论意义和应用价值。近年来，转录因子研究为人们认识果实成熟衰老机制开辟了新视野，逐渐成为国际研究热点。其中，MADS-box转录因子家族成员RIN最受关注。申请人在前期工作中，以番茄果实为材料，通过蛋白质组学技术结合染色质免疫共沉淀等方法，鉴定出RIN调控的5个新靶标基因，并首次证实RIN直接调节芳香物质代谢途径。本项目拟在已有的研究基础上，利用RNA-Seq技术分析rin突变株和野生型番茄果实在成熟衰老过程中的基因表达差异，挖掘RIN调控的下游基因，特别是转录因子，并通过分子生物学、生物化学、细胞生物学和植物生理学等实验方法，深入分析RIN调控的下游转录因子在果实成熟衰老中的重要作用，为进一步利用基因工程技术来控制果实成熟衰老。

Fruits are developmental structures that are unique to flowering plant and play a central role in seed maturation and dispersal. Ripening and senescence is a physiological feature of fruit. It is very important to explore the mechanisms regulating fruit ripening and senescence both in theoretical basis and practical implications. With many regulatory genes being identified in fruit, fruit-specific control of ripening and senescence by transcription factors has become an important research focus. Among them, transcription factor RIN has received more attention. In our previous study, using tomato as a model system, we have found five novel RIN-regulated direct target genes through the methods of proteomics-based chromatin immunoprecipitation, and firstly proved that aromatic metabolic pathway are directly regulated by RIN. Based on the previous work, this project is planed to use RNA-Seq technology to compare the gene expression of wild type and rin mutant for mining the downstream regulatory genes of RIN, especially transcription factors. Furthermore, we perform the methods of molecular biology, biochemistry, cell biology and plant physiology to explore the important roles of these transcription factors in fruit ripening and senescence, providing theory basis for engineering technology to delay fruit ripening and senescence.