功能丧失分析对于剖析生物过程的复杂性至关重要，包括配子发生。病毒诱导的基因沉默（VIGS）已被广泛用于作物物种，作为产生基因敲除的一种灵活且快速的方法。作为一种瞬时检测，VIGS通过费力且耗时的组织培养技术避免了稳定转基因系的产生。VIGS涉及在早期发育期间用携带内源基因靶序列的遗传操作的病毒构建体接种植物。病毒感染触发宿主植物基因沉默机制以将病毒基因组RNA加工成包括基因互补区的小RNA(sRNA)。具有与内源基因互补序列的sRNA介导靶基因的转录后基因沉默。这里,我们提供了一种简单且可重复的VIGS方案，该方案采用了番茄中的烟草摇铃病毒（TRV）（Solanumlycopersicumcv。M82).由于它在后期发育阶段是稳定的，因此该方法适用于番茄中的许多性状，包括配子发生，并且可以适用于其他作物物种。
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.
