TRV病毒介导的基因沉默体系在新疆陆地棉和亚洲棉中的建立

Establishment of TRV-Mediated Virus-Induced Gene Silencing in Xinjiang Upland Cotton and Asiatic Cotton

为研究烟草脆裂病毒(Tobacco rattle virus,TRV)介导的基因沉默(Virus-induced gene silencing,VIGS)技术应用的广谱性,以陆地棉(Gossypium hirsutum)CLA1为标记基因,通过构建基因沉默载体,在新陆早33和亚洲棉(Gossypium arboreum)建立了TRV-VIGS体系。半定量PCR分析表明,与对照相比,TRV侵染的新陆早33和亚洲棉中CLA1基因的表达均被有效抑制,真叶表现出明显的白化症状。同时在45份新陆中系列棉花材料中重复该体系,并且通过控制培养温度来检验温度对VIGS的影响,结果表明,当昼/夜温度分别为23℃/21℃和32℃/28℃时,不同品种的新陆中系列棉花均可以维持不同程度的白化表型。证明,TRV介导的VIGS可以用于室内研究新陆中系列棉花和亚洲棉基因功能。

To examine the application of a tobacco rattle vires （TRV）-mediated gene silencing （VIGS） technology in cotton varieties of the Xinluzhong series, we established a TRV-VIGS system for Xinluzao 33 and Gossypium arboreum using upland cotton GhCLA1 as a marker gene for silencing. Semi-quantitative real-time PCR analysis showed that GhCLA1 could be effectively silenced in Xinluzao 33 and G. arboreum. This system was further tested in 45 cotton varieties of the Xinluzhong series simultaneously. The results showed that after 2 weeks of inoculation, Xinluzao 33 and G. arboreum could maintain an albino phenotype to varying degrees at temperatures of 23℃/21℃ and 32℃/28℃ （day/night）, respectively. Our study suggests that the TRV-VIGS system can be widely applied in different cotton varieties of the Xinluzhong series and in Gossypium arboreum to help functionally characterize genes in the laboratory.