Abstract
As a promising high-throughput reverse genetic tool in plants, virus-induced gene silencing (VIGS) has already begun to fulfill some of this promise in diverse aspects. However, review of the technological advancements about widely used VIGS system, tobacco rattle virus (TRV)-mediated gene silencing, needs timely updates. Hence, this article mainly reviews viral vector construction, inoculation method advances, important influential factors, and summarizes the recent applications in diverse plant species, thus providing a better understanding and advice for functional gene analysis related to crop improvements.
Highlights
Virus-induced gene silencing (VIGS) is a high-throughput reverse genetics technique that exploits an RNA-mediated antiviral defense mechanism [post-transcriptional gene silencing (PTGS)] for functional gene analysis (Ratcliff et al, 1997; Sunilkumar et al, 2006)
We firstly summarize the development history of the vector construction and inoculation methodology, and focus on the significant efficiency influential factors in virus-induced gene silencing (VIGS) application, as well as the adoptable range of plant species of tobacco rattle virus (TRV)-VIGS is updated here
In the past two decades, many VIGS vectors have been developed (Supplementary Table 1), and TRV was preferentially used for VIGS assays in most dicots and some monocots, due to the high susceptibility of a wide range of hosts with mild viral symptoms after infection (Chen et al, 2005; Zhang et al, 2017; Xie et al, 2019)
Summary
Virus-induced gene silencing (VIGS) is a high-throughput reverse genetics technique that exploits an RNA-mediated antiviral defense mechanism [post-transcriptional gene silencing (PTGS)] for functional gene analysis (Ratcliff et al, 1997; Sunilkumar et al, 2006). In the following year, Liu et al (2002a,b) developed the most commonly used TRV vector (TRV2-MCS, pYL156; Figure 2C) by using the duplicated CaMV 35S promoter (2 × 35S), instead of a single 35S, and adding a self-cleaving ribozyme (Rz, to increase virus infectivity) before the nopaline synthase terminator (NOSt) on the transferred Agrobacterium tumefaciens T-DNA of plant binary transformation vectors, which had a high silencing efficiency (90–97.9%) They successfully studied the function of tobacco RAR1 and other genes against TMV resistance (Liu et al, 2002a,b; Tian et al, 2014). More positive controls should be found and determined for some specific regions in VIGS system, such as root or other organs
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