Post-transcriptional Gene Silencing Using Virus-Induced Gene Silencing to Study Plant Gametogenesis in Tomato.
Post-transcriptional Gene Silencing Using Virus-Induced Gene Silencing to Study Plant Gametogenesis in Tomato.
复制标题
转录后基因沉默利用病毒诱导的基因沉默来研究番茄植物配子发生。
DOI：
10.1007/978-1-0716-2253-7_15
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发表时间：
2022
期刊：
影响因子：
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通讯作者：
Wang Z
中科院分区：
文献类型：
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作者：
Wang Z
功能丧失分析是剖析包括配子发生在内的生物过程的复杂性质所必需的。病毒诱导的基因沉默(VIGS)作为一种顺从的、快速的基因敲除方式在农作物中得到了广泛的应用。作为一种瞬时检测方法，VIGS通过费时费力的组织培养技术避开了稳定转基因品系的产生。VIGS涉及在植物发育的早期用携带内源基因靶标序列的基因操纵的病毒构建物接种植物。病毒感染触发宿主植物的基因沉默机制，将病毒基因组RNA加工成包括基因互补区的小RNA(SRNAs)。与内源基因互补的sRNAs介导靶基因的转录后基因沉默。在这里，我们提供了一种简单且可重现的VIGS方案，该方案利用烟草Rattle病毒(TRV)在番茄(Solanum lycopsicum cv.M82)。由于它在发育后期是稳定的，因此该方法适用于番茄的许多性状，包括配子发生，并且它也可以适应其他作物。
Loss-of-function analyses are essential to dissect the complex nature of biological processes, including gametogenesis. Virus-induced gene silencing (VIGS) has been widely used in crop species as an amenable and rapid way to generate gene knockdowns. As a transient assay, VIGS circumvents the generation of stable transgenic lines through laborious and time-consuming tissue culture techniques. VIGS involves inoculating plants during early development with genetically manipulated viral constructs carrying an endogenous gene target sequence. The viral infection triggers the host plant gene silencing machinery to process the viral genomic RNA into small RNAs (sRNAs) including the gene complementary region. The sRNAs with complementary sequences to the endogenous gene mediate posttranscriptional gene silencing of the targeted gene. Here, we provide a simple and reproducible VIGS protocol employing the tobacco rattle virus (TRV) in tomato (Solanum lycopersicum cv. M82). As it is stable at later developmental stages this approach is suitable for many traits in tomato including gametogenesis and it can be adapted to other crop species.