菊花CmTCP7转录因子调控开花的分子机制研究

菊花是我国十大传统名花和世界四大切花之一，花期是其重要观赏性状。目前多采用促成或抑制栽培调控其花期以满足周年供应，存在成本高、品质难以保障等不足。解析菊花开花分子机制是其花期遗传改良的关键。项目组发现TCP家族成员CmTCP7具有调控菊花开花的功能。然而，TCP7调控植物开花分子机制尚未见报道，为此，本申请项目拟借助酵母单杂交发掘CmTCP7上游转录因子，并借助EMSA、菊花原生质体瞬时表达荧光素系统、ChIP技术进行验证；采用酵母双杂交发掘CmTCP7互作蛋白，并借助BiFC、GST Pull-down、CoIP技术进行验证；借助反向遗传学方法对关键上游转录因子、互作蛋白基因进行功能鉴定；对CmTCP7超表达及沉默转基因植株进行转录组分析，解析CmTCP7调控的下游开花路径；从而阐明CmTCP7调控菊花开花分子机制。项目的实施，不仅可揭示菊花开花新分子网络，亦可丰富植物开花分子机理。

Chrysanthemum is one of ten traditional Chinese famous flowers and one of four cut flowers in the world. Flowering time is an important trait of chrysanthemum. To meet the need of year-round production, accelerating or delaying cultivation practices have been employed to modulate the flowering time of chrysanthemum, which increased the cost of produce to great extend, and hampered the quality of the product as well. Molecular mechanisms of flowering time is important for genetic engineering of flowering time of chrysanthemum. We have previously found that CmTCP7 played an important role in regulating flowering time of chrysanthemum, however, the mechanisms of TCP7 in regulating flowering time in plant has not been clarified yet. In present proposal, we aimed to: 1) screen upstream transcription factor of CmTCP7 and validate the binding activity of upstream TF with the promoter of CmTCP7 by Eletrophoretic Mobility Assay (EMSA), Chromatin Immunoprecipitation (ChIP) and Fluorescein transient expression assay using chrysanthemum protoplast; 2) screen the interaction proteins with CmTCP7 by yeast two hybrid, and to validate the interaction via Bimolecular fluorescence complementation (BiFC), GST Pull down and Co-Immunoprecipitation (CoIP), 3) investigate the flowering time gene regulated by CmTCP7 via RNA-Seq. and 4) identify the function of the discovered important upstream TFs and genes encoding interaction proteins via reverse genetic approaches. Based on the above evidences, the regulatory roles of the CmTCP7 in flowering time could be clarified. The complement of the proposal would make a breakthrough not only in the molecular mechanisms of flowering time of chrysanthemum but also would be a good add to those of other plants.