Virus-induced gene silencing (VIGS) is a technology that exploits an RNA-mediated antiviral defense mechanism. In plants infected with unmodified viruses the mechanism is specifically targeted against the viral genome. However, with virus vectors carrying inserts derived from host genes the process can be additionally targeted against the corresponding mRNAs. VIGS has been used widely in plants for analysis of gene function and has been adapted for high-throughput functional genomics. Until now most applications of VIGS have been in Nicotiana benthamiana. However, new vector systems and methods are being developed that could be used in other plants, including Arabidopsis. Here we discuss practical and theoretical issues that are specific to VIGS rather than other gene "knock down" or "knockout" approaches to gene function. We also describe currently used protocols that have allowed us to apply VIGS to the identification of genes required for disease resistance in plants. These methods and the underlying general principles also apply when VIGS is used in the analysis of other aspects of plant biology.
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"In this article, we report the use of a virus-induced gene-silencing (VIGS)-based, fast-forward genetics approach (Baulcombe, 1999; Lu et al., 2003b; del Pozo et al., 2004) to identify plant genes that play a role in nonhost disease resistance. One of the genes identified through the screen encodes the photorespiratory enzyme , GOX. "
"Virus-induced gene silencing (VIGS), a technique developed for gene transcript suppression, has been used for the characterization of the function of plant genes and avoids the disadvantages of regenerating plants from difficult species ( Burch-Smith et al. 2004, Hileman et al. 2005, Wege et al. 2007, Senthil-Kumar and Mysore 2011, Ramegowda et al. 2014). Virus-induced gene silencing has been widely used for functional gene analysis in species like Nicotiana benthamiana (Solanaceae) ( Ratcliff et al. 2001, Lu et al. 2002), A. thaliana (Brassicaceae) ( Burch-Smith et al. 2006, Wang et al. 2006), tomato (Solanaceae) ( Liu et al. 2002a, Ekengren et al. 2003), barley (Gramineae) ( Holzberg et al. 2002), Papaver somniferum (Papaveraceae) ( Hileman et al. 2005), Phalaenopsis amabilis (Orchidaceae) ( Lu et al. 2007), Aquilegia vulgaris and Thalictrum sp. (Ranunculaceae) ( Gould and Kramer 2007, Kramer et al. 2007, Di Stilio et al. 2010), Spinacea oleracea (Amaranthaceae ) ( Golenberg et al. 2009), Jatropha curcas (Euphorbiaceae ) ( Ye et al. 2009), Zingiber officinale (Zingiberaceae) ( Renner et al. 2009), and Malus domestica and Pyrus pyrifolia (Rosaceae) ( Sasaki et al. 2011). "
"These are detected by the plant host and trigger PTGS, which involves the generation of small interfering RNAs (siRNAs) that are then loaded into the RNA-induced silencing complex containing at least one Argonaute endonuclease to guide the cleavage of complementary viral RNA (Baulcombe, 2004). In VIGS, a fragment of a plant gene is inserted into a virus vector to form a recombinant virus that, upon infection of a plant host, induces PTGS targeting both the virus RNA and homologous endogenous plant RNA sequences for degradation (Lu et al., 2003). Only one plant virus, Barley stripe mosaic virus (BSMV), which has a tri-partite RNA genome comprising RNAa, RNAb and RNAc, has so far been developed into a VIGS vector for wheat (Scofield et al., 2005). "