lncRNA与miRNA协同调控番茄晚疫病的分子机制研究

Tomato late blight (LB) is one of frequent and multiple diseases, which causes severe loss of tomato production and the major economic. So it is very urgent and significant for the study on tomato resistance mechanism against pathogen causing LB and exploration of effective control measures. In recent years, we have continuously identified a number of miRNAs and lncRNAs related to tomato LB, studied their function and found the signal pathways associated with them, showing that the relationship between lncRNA and miRNA is promoted or inhibited each other. In this project, we will identify the differentially expressed lncRNAs by constructing a variety of transcriptome libraries, explore their relationship with miRNAs, construct regulated network including lncRNA, mRNA and miRNA, clone the key target genes, and identify the functions by transgenic approach. We will select pathogen-resistance tomato plants by inoculating pathogen into the transgenic plants, while the regulated network will be improved and this will further reveal the mechanism of lncRNA and miRNA synergistic regulation of tomato LB. These will provide not only theoretical basis and technical support for tomato disease-resistant breeding, but also the consultation for the study of other plant non-coding RNAs.

晚疫病是一种常发性、多发性病害，对番茄产量、品质及经济效益影响甚大，掌握其调控的分子机理，寻求有效的治理措施至关重要。近年来，我们接连不断地发现并鉴定了对番茄晚疫病有明显调控作用的一系列miRNA、lncRNA的功能以及它们参与的信号通路；初步预测到部分lncRNA与miRNA存在相互抑制或促进的关系。本项目拟通过全转录组测序，进一步筛选差异表达的lncRNAs；探寻它们与miRNAs的作用效果；构建lncRNA、miRNA、mRNA彼此间的调控网络；克隆网络节点上重要的基因；利用过表达和沉默技术手段获得相应的转化株，接种病原物检测分析各株系的抗性；在验证这些基因功能、选出抗病新种质的同时，完善该网络并揭示lncRNA与miRNA协同调控晚疫病的机制。为番茄抗病、育种提供理论基础和技术支持；也为其他植物非编码RNA的研究提供参考。

晚疫病对番茄的产量、品质及经济效益影响甚大，在世界各地的番茄产区，每年都有不同程度的发生。查明病害的调控机理，寻求有效的防治策略是亟待解决的问题。本项目在挖掘主要调控番茄晚疫病的非编码RNA的同时，探究部分lncRNA、miRNA、mRNA等构成的网络对晚疫病的调控机制以及网络节点上各关键基因的功能。发现了miR1916通过靶向晚疫病抗性蛋白同源物R1B-16、抗病蛋白RPP13、MYB12基因等，导致α-番茄碱和花青素的积累，从而负调控番茄对晚疫病的抗性；揭示了WRKY1通过启动lncRNA33732，诱导RBOH表达，促进H2O2的生成，从而增强番茄抗性的机制；阐明了番茄lncRNA15492通过与miR482a前体及成熟体间的相互作用，调控番茄体内NBS-LRR的稳态，进而影响番茄抗病性的分子机制；发现了番茄抗晚疫病中起负调控作用的lncRNA39896，通过吸附miR166b影响其下游靶基因HD-Zip转录因子的表达，形成lncRNA39896–miR166b–HDZs模块来调控番茄对晚疫病的抗性。该项目取得的成果为揭示番茄抗晚疫病的机制以及植物非编码RNA的调控作用提供了新的理论和思路。在该项目的支持下，发表学术论文35篇，其中2篇入选ESI高被引；申请发明专利4件（已授权1件）；培养了4名博士、6名硕士；参加国内外学术会议18次，8次做了口头报告。