不同温度下CsbHLH14选择性剪切调控茶树单萜类香气物质生物合成的分子机理

单萜类化合物是决定茶叶香气品质特征的主要茶树次生代谢产物之一，揭示单萜类的生物合成及调控机理对于提高成品茶的香气品质至关重要。申请人前期研究表明：茶树转录因子CsbHLH14可能受温度影响发生选择性剪切，进而参与单萜类香气物质的调控。本项目拟采用RT-qPCR和GC-MS技术，分析CsbHLH14选择性剪切产生的转录本与单萜类积累以及温度变化的关系；通过在拟南芥中过表达和茶树叶片中抑制表达CsbHLH14剪切体，研究CsbHLH14不同转录本的功能；利用染色质免疫共沉淀分离更多与单萜合成相关的CsbHLH14靶基因。研究结果将有助于解析CsbHLH14在茶树单萜类香气物质生物合成中的功能，揭示基因与环境、基因与基因网络调控茶树单萜类香气物质生物合成的机理，为最终实现茶叶香气品质调控奠定基础。

Monoterpenoids are the most important secondary metabolites in tea plant (Camellia sinensis (L.) O. Kuntze) which determine the characteristics of tea product. Illustrating the biosynthesis and regulation mechanism of monoterpenoids is beneficial for improving the aroma of tea. In our previous study, a temperature-responsive transcription factor CsbHLH14 was found to undergo alternative splicing events, which may regulate the monoterpenoid biosynthesis in tea plants. In this study, the interaction relationships among transcripts produced by alternative splicing, monoterpenoids accumulation and temperature changes will be investigated via RT-qPCR and GC-MS. The function of CsbHLH14 transcripts in regulating monoterpenoids biosynthesis will be validated using over-expression in Arabidopsis thaliana and transient suppression expression in tea leaves. Moreover, the target genes of CsbHLH14 related to monoterpene biosynthesis will be explored by Chromatin Immuno-precipitation. The results will contribute to elucidate the function of CsbHLH14 in regulating monoterpenoids biosynthesis, and will uncover the regulatory mechanisms of gene networks and environments regulating monoterpenoids biosynthesis, which can provide a theoretical foundation for promoting the aroma quality in tea.