陆地棉抗黄萎病相关基因GhRPL18的分析与功能验证

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• 英文论文题名：Analysis and Functional Verification of the Verticillium Wilt-Related Gene GhRPL18 in Upland Cotton(Gossypium hirsuturm L.)
• 论文作者：龚骞 ; 杨召恩 ; 杨作仁 ; 李付广
• 英文论文作者：Gong Qian ; Yang Zhao'en ; Yang Zuoren ; Li Fuguang ; Institute of Cotton Research of Chinese Academy of Agriculture Sciences/State Key Laboratory of Cotton Biology
• 年：2017
• 作者机构：中国农业科学院棉花研究所/棉花生物学国家重点实验室;
• 论文关键词：陆地棉 ; 大丽轮枝菌 ; RPL18 ; qRT-PCR ; VIGS
• 英文论文关键词：upland cotton ; Verticillium dahliae ; RPL18 ; real-time quantitative reverse-transcription polymerase chain reaction ; virus induced gene silencing
• 会议召开时间：2017-08-07
• 会议录名称：中国农学会棉花分会2017年年会暨第九次会员代表大会论文汇编
• 语种：中文
• 分类号：S435.621.2
• 学会代码：MHXB
• 会议名称：中国农学会棉花分会2017年年会暨第九次会员代表大会
• 会议地点：中国河南郑州
• 主办单位：中国农学会棉花分会
• 学会名称：《棉花学报》编辑部
• 页数：1
• 文件大小：61k
• 原文格式：O
• 会议级别：全国

摘要

基于中植棉2号鉴定出与抗黄萎病相关的核糖体蛋白基因,命名为GhRPL18。A组该基因全长1397 bp,D组中其全长1387 bp;两者的编码序列(CDS)区均为537 bp,编码178个氨基酸。进化树分析结果表明GhRPL,18蛋白与Theobroma cacao(可可)中的RPL18蛋白的相似性最高,Weblogo分析结果表明该蛋白的保守性很强。通过对A组和D组中GhRPL18分析,发现两者相似性极高,且启动子区有与抗病相关的大量顺式作用元件。qRT-PCR结果表明GhRPL18在抗病品种(中植棉2号)中被黄萎病菌诱导表达,而在感病品种(冀棉11)中表达量变化不大。通过病毒诱导基因沉默(VIGS)技术,在中植棉2号中成功沉默基因GhRPL18;利用表型观察、统计病指、大丽轮枝菌定量以及恢复实验,我们发现抗病品种中的GhRPL18被沉默后,其对黄萎病的抗性丧失,这表明GhRPL18在陆地棉抗黄萎病中起到了重要作用。
A new gene,GhRPL18,was identified from cotton cultivar Zhongzhimian 2,which was resistan to Verticilhum wilt.The length of the gene was 1397 bp in group A and 1387 bp in group D.The coding sequence region was 537 bp and encoded 178 amino acids.The results of phylogenetic tree analysis showed that the similarity between GhRPL18 protein and RPL18 protein in Theobroma cacao(cocoa) was the highest.The results of Weblogo analysis showed that the protein was highly conserved.By analyzing GhRPL18 in group A and group D,we found that the similarity between them was high and that the promoter region had a large number of cis-acting elements associated with disease resistance.Furthermore,the result of qRT-PCR showed that GhRPL18 was induced by Verticillium dahliae in the resistant cultivar(Zhongzhimian 2).However,the expression of GhRPL18 in the susceptible variety(Jimian 11) remained constant.We found that the resistant cultivar became susceptible to infection by constructing virus induced gene silencing interference vector to interfere GhRPL18 in resistant cultivar.Furthermore,statistical analysis of disease,quantitative simulation of Verticillium dahliae,and the recovery experiment also confirmed these results.This suggests that GhRPL18 plays an important role in the resistance to the infection of Verticillium dahliae in upland cotton.

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