番茄果实成熟相关外显子环化RNA（circRNA）的鉴定和初步功能分析

项目名称： 番茄果实成熟相关外显子环化RNA（circRNA）的鉴定和初步功能分析

项目编号： No.31501766

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 农业科学

项目作者： 谭金娟

作者单位： 浙江省农业科学院

项目金额： 21万元

中文摘要： 番茄是重要的蔬菜作物。作为研究果实成熟的模式植物，其果实成熟机理在分子和遗传水平已有深入研究。乙烯调控番茄果实成熟，但乙烯依赖和非依赖途径中，相关基因如何互作并调控成熟的某一具体生理现象仍需进一步阐述。外显子环化RNA（circRNA）是近年来被证明参与基因表达调控的重要非编码RNA成员，其在植物中的研究极少。通过环形RNA高通量测序，我们从番茄果实中检测出数千个潜在circRNA位点，并用分子手段验证了部分位点，包括来源于成熟调控基因（ACO2和ETR4）的6个circRNA。本项目拟开展番茄成熟（乙烯依赖和非依赖途径）相关circRNA的功能研究：（1）进一步挖掘成熟调控和/或乙烯敏感突变体Nr相关的circRNA；（2）分子鉴定一批所发现circRNA；（3）利用过表达和抑制表达，初步解析2-4个乙烯和成熟相关circRNA的功能。本研究将进一步丰富果实成熟和乙烯调控的分子机理。

中文关键词： 番茄；果实；成熟；乙烯；外显子环化RNA

英文摘要： Tomato is an economic important vegetable crop worldwide. As the model system for studying fruit ripening, tomato fruit ripening has been characterized in detail at the molecular and genetic levels. Ethylene regulates tomato fruit ripening, but how ethylene-dependent and -independent genes interact to control specific physiological aspects of ripening remains to be further characterized. Exonic circular RNAs (circRNAs) are a class of newly characterized non-coding RNAs that regulate gene expression in humans and animals, but their function in growth and development of plants are poorly characterized. By using high-throughput circular RNA sequencing and bioinformatic analysis, we have identified thousands of putative circRNA sites from tomato fruits at different ripening stages. In addition, a number of circRNAs were confirmed by sequencing and RNase-R protection assay, including six circRNAs originated from ripening regulator ACO2 and ETR4. In this proposal, molecular and functional characterization of circRNAs involved in ethylene-dependent and –independent tomato fruit ripening will be performed: (1) further identification of putative circRNA sites involved in tomato fruit ripening and/or regulated by Never-ripening mutant (Nr); (2) molecular characterization of a large number of circRNAs associated with fruit ripening and/or Nr mutant; (3) functional analysis of two to four circRNAs originated from ethylene and ripening associated genes by over-expression and suppress-expression in tomato. This study will further our understanding of fruit ripening and ethylene physiology at the molecular levels.

英文关键词： tomato;fruit ;ripening;ethylene;circRNA