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。
[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.
作者
黄俊森
王星星
杨力
裴小雨
贺昆仑
刘艳改
张飞
张先亮
王军
马雄风
杨代刚
高俊山
Huang Junsen;Wang Xingxing;Yang Li;Pei Xiaoyu;He Kunlun;Liu Yangai;Zhang Fei;Zhang Xianliang;Wang Jun;Ma Xiongfeng;Yang Daigang;Gao Junshan(School of Life Sciences,Anhui Agricultural University,Hefei 230000,China;Institute of Cotton Research of Chinese Academy of Agricultural Sciences/State Key Laboratory of Cotton Bio-breeding and Integrated Utilization/Key Laboratory for Cotton Genetic Improvement,Ministry of Agriculture and Rural Affairs,Anyang,Henan 455000,China)
出处
《棉花学报》
CSCD
北大核心
2024年第3期211-224,共14页
Cotton Science
基金
棉花生物育种与综合利用全国重点实验室自主课题:单细胞多组学解析棉花黄萎病防御机制(CB2021E02)
新疆维吾尔自治区重点研发任务专项:“双30A”早熟机采棉品种选育与配套技术研究(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:多耐受性高品质棉花新品种选育及配套栽培技术示范推广(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。
