茶树低温胁迫相关micRNA分离鉴定及与其靶标基因调控互作机制研究

Since low temperature is one of the major factors affecting tea quality and production, underlying the mechanism of resisting to low temperature in tea plant (Camellia sinensis L.) warrants serious attention. Based on our solid stress-response research which has been developed in the past few years, this project aims to study the expression and regulation of related microRNA and function of their target genes under cold stress in tea plant. Candidate microRNA response to cold stress will be identified with deep sequencing of microRNA and microarray technology. By degradation sequencing, target genes will be precisely sequenced. Action mode of microRNA with significant differential expression and the target genes will be revealed by real-time quantitative PCR. The functions of the target genes will be analyzed by functional complementary experiment using Arabidopsis mutants. This project will be of critical importance to understand the cold stress related microRNA in tea plant as well as expression and regulation mechanism of their target gene, and provide possible basis for the further study of microRNAs in regulating cold resistance in tea plant.

低温是影响茶叶产量和品质的重要因素之一， 茶树抗寒机制的研究意义重大。为促进茶树抗寒分子生物学研究发展，本课题组前期已通过高通量microRNA测序及微阵列定制microRNA芯片杂交技术，获得与茶树低温胁迫响应相关的microRNA。为进一步揭示microRNA与其靶基因之间调控互作机制及其在茶树抗寒性状过程中的重要功能，本项目拟在已有的研究基础上，将利用降解组测序技术，对靶基因进行准确预测；选取显著差异表达的microRNA及其对应的靶基因分别设计引物进行real-time荧光定量PCR实验，分析两者的作用机理；并利用拟南芥突变体基因功能互补实验来研究靶基因的主要功能特点及其作用时空和方式。本课题对认识茶树低温胁迫相关microRNA及其靶基因之间互作调控机制有重要重要意义，是进一步利用miRNA调控茶树抗寒性，选育优良茶树抗寒品种的科学基础。

低温是影响茶叶产量和品质的重要因素之一， 茶树抗寒机制的研究意义重大。为促进茶树抗寒分子生物学研究发展，本课题组已通过高通量microRNA测序及微阵列定制microRNA芯片杂交技术，获得与茶树低温胁迫响应相关的microRNA。为进一步揭示microRNA与其靶基因之间调控互作机制及其在茶树抗寒性状过程中的重要功能，本项目在已有的研究基础上，利用降解组测序技术，对靶基因进行准确预测；选取显著差异表达的microRNA及其对应的靶基因分别设计引物进行real-time荧光定量PCR实验，分析两者的作用机理；并利用拟南芥突变体基因功能互补实验来研究靶基因的主要功能特点及其作用时空和方式。本课题对认识茶树低温胁迫相关microRNA及其靶基因之间互作调控机制有重要意义，是进一步利用miRNA调控茶树抗寒性，选育优良茶树抗寒品种的科学基础。