Gh_D11G050000参与棉花黄萎病抗性的功能分析
Functional analysis of Gh_D11G050000involved in cotton resistance to Verticillium wilt
黄俊森 1王星星 2杨力 2裴小雨 2贺昆仑 2刘艳改 2张飞 2张先亮 2王军 3马雄风 2杨代刚 2高俊山3
作者信息
- 1. 安徽农业大学生命科学学院,合肥 230000;中国农业科学院棉花研究所/棉花生物育种与综合利用全国重点实验室/农业农村部棉花生物学与遗传育种重点实验室,河南 安阳 455000
- 2. 中国农业科学院棉花研究所/棉花生物育种与综合利用全国重点实验室/农业农村部棉花生物学与遗传育种重点实验室,河南 安阳 455000
- 3. 安徽农业大学生命科学学院,合肥 230000
- 折叠
摘要
[目的]克隆Gh_D11G050000基因,解析其在棉花黄萎病抗性反应中的功能和作用机制,为开展棉花抗病分子育种提供理论依据.[方法]利用生物信息学方法分析Gh_D11G050000基因的序列特征和系统进化关系.通过实时荧光定量聚合酶链式反应(quantitative real-time polymerase chain reaction,qRT-PCR)分析该基因的表达模式以及大丽轮枝菌(Verticillium dahliae)侵染后的表达量变化.利用病毒诱导的基因沉默(virus-induced gene silencing,VIGS)技术初步验证该基因的功能.通过转录组测序分析和相关基因的表达量测定初步解析其抗病机制.[结果]Gh_D11G050000 与Gh_A11G049600 和Gr_11G034620 蛋白的亲缘关系最近.qRT-PCR分析发现,Gh_D11G050000在棉花根部的表达量最高;该基因的表达量在大丽轮枝菌侵染后显著增加.Gh_D11G050000沉默植株对大丽轮枝菌的抗性减弱,具体表现为茎秆维管束褐变加重,有病菌繁殖的茎段数量明显增加,病株率和病情指数显著升高.转录组及qRT-PCR分析表明,Gh_D11G050000沉默棉株中茉莉酸、乙烯信号通路和木质素合成途径中相关基因的转录水平降低.[结论]Gh_D11G050000通过影响木质素合成基因、茉莉酸和乙烯信号通路相关基因的表达正调控棉花对黄萎病的抗性.
Abstract
[Objective]This study aims to provide a theoretical basis for molecular breeding of disease-resistant cotton by cloning Gh_D11G050000 gene,and to analyze its function and mechanisms in cotton resistance to Verticillium wilt.[Methods]Bioinformatic method was utilized to analyze the sequence characteristics and phylogenetic relationship of Gh_D11G050000.Quantitative real-time polymerase chain reaction(qRT-PCR)was employed to investigate the expression pattern of this gene,and it's expression level changes after infected by Verticillium dahliae.The function of this gene was validated using virus-induced gene silencing(VIGS)technique.Transcriptome sequencing analysis and the detection of relative expression levels of related genes were conducted to explore the mechanisms of disease resistance.[Results]Gh_D11G050000 exhibited close relationship with Gh_A11G049600 and Gr_11G034620 protein.qRT-PCR analysis revealed that Gh_D11G050000 was highly expressed in cotton roots,and the expression level of Gh_D11G05000 was significantly increased after V.dahliae infection.Gh_D11G050000 silenced cotton plants showed reduced resistance against V.dahliae,characterized by aggravated browning of stem vascular bundles,increased the number of stem segments with bacterial propagation,as well as significantly increased the rate of diseased plants and disease index.Transcriptome analysis combined with qRT-PCR demonstrated that Gh_D11G050000 silenced cotton plants showed decreased transcription levels for several genes in jasmonic acid(JA)signaling pathway,ethylene(ET)signaling pathway,and lignin synthesis pathway.[Conclusion]Gh_D11G050000 positively regulates cotton resistance to Verticillium wilt by influencing the expression of several genes involved in lignin synthesis,JA,and ET signaling pathways.
关键词
陆地棉/黄萎病/Gh_D11G050000/抗病基因/病毒诱导的基因沉默/茉莉酸/乙烯/木质素Key words
upland cotton/Verticillium wilt/Gh_D11G050000/resistance gene/virus-induced gene silencing/jasmonic acid/ethylene/lignin引用本文复制引用
基金项目
棉花生物育种与综合利用全国重点实验室自主课题(CB2021E02)
新疆维吾尔自治区重点研发任务专项(2022B02052-2)
新疆维吾尔自治区昌吉回族自治州科技支撑产业高质量发展专项(2021Z01-01)
国家棉花产业技术体系中早熟品种改良岗位科学家(CARS-15-07)
新疆维吾尔自治区棉花产业技术体系资源研究与育种(早熟棉)岗位专家()
出版年
2024
国家科技期刊平台
NSTL
万方数据