乙烯响应因子ERF调控番茄不规则裂果的机理解析

裂果严重影响番茄果实品质，制约番茄产业发展。但番茄裂果机理研究滞后。前期研究发现，乙烯响应因子（ERF）是调控番茄不规则裂果的关键基因之一，但ERF响应裂果的分子机理及调控网络尚未明晰。为此，申请者拟克隆调控番茄裂果关键ERF的基因全长并进行启动子分析，通过酵母单杂交筛选其上游调控因子，采用荧光素酶瞬时表达、EMSA等技术进行验证，明确ERF上游的调控因子并对其功能进行验证；通过表达特性、亚细胞定位明确ERF的基本特性，并利用超表达、基因沉默技术获得ERF的转基因番茄植株，进一步对番茄转基因株系及野生型进行转录组分析，明确ERF调控番茄裂果的机理及下游调控网络。结合申请者前期对耐裂果及易裂果材料的转录、翻译组测序和BSA-seq分析，揭示ERF调控番茄裂果的分子机理和调控网络，明确其关键调控单元。该研究一方面可丰富裂果的理论基础，另一方面可为番茄耐裂果性状的遗传改良提供有价值的基因资源。

Cracking seriously influences fruit quality and will then restrict the development of the tomato industry. However, the molecular mechanism of tomato cracking is lagging behind its development. Previous studies have shown that ethylene-responsive factor(ERF) was key factor related to fruit irregular cracking, but its molecular mechanism and regulatory network are still unclear. In this research, we will clone the key ERF that influences fruit irregular cracking followed by promoter analysis. Yeast one-hybrid will then be carried out to identify its upstream regulator. And in vivo luciferase transient expression and in vitro EMSA assay will be used to confirm its regulatory roles. Next, the key genes' expression profiles, subcellular localization, and generating its overexpression or gene silencing techniques will be adopted to verify the gene function. And transcriptional sequencing will be carried out with transgenic and wild-type plants to explore the downstream regulatory network. Combined with our previous transcriptome sequencing, ribo-seq and BSA-seq analysis, the current project will provide clues for illustrating the molecular mechanisms of ERF, regulatory network and key regulatory unit of fruit cracking, which can further our knowledge on fruit cracking. On the other hand, potential elite genes that can be used for cracking-oriented genetic engineering of tomato will be exploited.