番茄长链非编码RNA LeLNR1在抗TYLCV中的作用及抗凋亡机制解析

项目名称： 番茄长链非编码RNA LeLNR1在抗TYLCV中的作用及抗凋亡机制解析

项目编号： No.31471873

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 余文贵

作者单位： 江苏省农业科学院

项目金额： 85万元

中文摘要： 长链非编码RNA（lncRNA）在生物发育以及免疫等过程中发挥重要作用。而番茄lncRNA的研究目前未见报道。本实验室发现一个番茄lncRNA（LeLNR1）在含Ty-2抗性位点的番茄中受番茄黄化曲叶病毒（TYLCV）诱导表达，将LeLNR1瞬时沉默后，番茄出现细胞凋亡和叶片卷曲，并且对TYLCV的抗性显著降低；LeLNR1在烟草中瞬时表达可抑制小鼠BAX蛋白引起的细胞凋亡。本项目拟通过转基因过量表达和RNAi研究LeLNR1在抗TYLCV和生长发育中的作用；通过RNA原位杂交明确其组织定位；将LeLNR1分段缺失明确其抗凋亡的核心区；转录组测序和Small RNA测序预测LeLNR1调控的靶基因，并结合VIGS瞬时沉默验证候选基因；通过RNA-Protein Pull Down分离与LeLNR1结合的蛋白；最终解析LeLNR1在调控番茄抗TYLCV中的作用和抑制细胞凋亡的机制。

中文关键词： 番茄；长链非编码RNA；抗凋亡；黄化曲叶病毒

英文摘要： Long non coding RNA (lncRNA) plays important roles in biological development and immunity, but there is none report about lncRNA in tomato until now. we have identified a tomato lncRNA (LeLNR1) which can be induced in the resistant lines with Ty-2 loci by the infection of yellow leaf curl virus (TYLCV). The plants with LeLNR1 silenced transiently showed apoptosis and leaf curling phenomenon, accompanyed with reduced resistance to TYLCV.Overexpression of LeLNR1 in tobacco can inhibit the apoptosis caused by the mice BAX protein.The research proposed to verify the function of LeLNR1 in the TYLCV resistance and growth development by transgenic research with it's overexpression in susceptible tomato and gene scilenced by RNAi in resistant tomato. RNA in situ hybridization was utilized to assure tissue expression pattern of LeLNR1.The core region determining anti apoptosis was identified by deletion analysis. The target genes of LeLNR1 would be predicated by analyzing transcriptome sequencing and Small RNA sequencing, combined with VIGS methods. The protein interacted with LeLNR1 would be obtained by RNA-Protein Pull Down. The aim is to prove the TYLCV resistance of LeLNR1 and it's mechanism of inhibiting apoptosis.

英文关键词： Tomato;long non coding RNA;anti-apoptosis;TYLCV