番茄抗病相关的miPEPs挖掘及功能研究

番茄生长发育过程中不可避免地要与病原体进行博弈，掌握并利用其自身潜能使之战胜异己对蔬菜生产至关重要。因此，亟待揭示抗病的机制。非编码RNA强大的调控作用被世人瞩目，但对其可编码小肽进而行使功能这一新兴前沿领域却知之甚少。本项目在较全面地掌握了番茄miR482、miR1916、miR172等的抗病调控作用，且实现了小肽对番茄抗性影响的系统研究工作基础上，拟将生物信息学分析与RACE实验等技术相结合，从野生番茄中挖掘抗病相关miRNAs的初级体（pri-miRNA）编码的小肽（miPEPs）；查明喷施处理对植株的影响；克隆主要基因，分别构建过表达载体和敲除载体，转化番茄创制新种质，检测再生植株的生理生化特性及抗性，明确各miPEP的功能；揭示各miPEP、miRNA及其靶基因间的协同调控关系。为番茄病害的有效治理提供理论和材料基础；也为其他植物的抗性研究及拓展非编码RNA的调控作用提供参考。

Tomato inevitably plays an interaction with pathogens during its growth and development. Mastering and using its own potential to defense is vital to vegetable production. So it is important to reveal the mechanism of disease resistance. Although the powerful regulatory effect of non-coding RNA has been widely documented, a few is known about the role of the non-coding RNA-encoded small peptide with strong regulatory effects as the emerging frontier research. Based on the comprehensive understanding of the regulation roles of disease resistance of tomato miR482, miR1916 and miR172, we discovery that some small peptides can enhance tomato resistance. This project aims to combine bioinformatics analysis, RACE experiments and other technologies to mine the small peptides (miPEPs) encoded by the primary miRNA (pri-miRNA) disease resistance-related in wild tomato and identify the effects of spraying on plants. Key genes are cloned and used to construct the overexpression vectors and then knockout vectors, transform tomato to create new germplasms. To detect the physiological and biochemical characteristics of transgenic tomato plants, analyze the resistance of plants to pathogen, and clarify the roles of each miPEP. Based on the above, the synergistic regulatory relationship between each miPEP, miRNA and its target genes are revealed. It provides a theoretical and material basis for the effective management of tomato diseases. It also provides a reference for the resistance research of other plants and the expansion of the regulatory mechanism of non-coding RNA.