Potato Virus X-Induced Gene Silencing in Leaves and Tubers of Potato

Abstract

Virus induced gene silencing (VIGS) is increasingly used to generate transient loss-of-function assays and has potential as a powerful reverse-genetics tool in functional genomic programs as a more rapid alternative to stable transformation. A previously described potato virus X (PVX) VIGS vector has been shown to trigger silencing in the permissive host Nicotiana benthamiana. This paper demonstrates that a PVX-based VIGS vector is also effective in triggering a VIGS response in both diploid and cultivated tetraploid Solanum species. We show that systemic silencing of a phytoene desaturase gene is observed and maintained throughout the foliar tissues of potato plants and was also observed in tubers. Here we report that VIGS can be triggered and sustained on in vitro micropropagated tetraploid potato for several cycles and on in vitro generated microtubers. This approach will facilitate large-scale functional analysis of potato expressed sequence tags and provide a noninvasive reverse-genetic approach to study mechanisms involved in tuber and microtuber development.

The Transcriptional Activator Pti4 Is Required for the Recruitment of a Repressosome Nucleated by Repressor SEBF at the PotatoPR-10aGene

“…Potato plants were propagated in vitro as described (Faivre-Rampant et al, 2004), but with some modifications. Five stem pieces per Magenta box were cultivated in 100 mL of MS medium (Murashige and Skoog, 1962) supplemented with 0.4 mg/mL thiamine, 0.5 mg/mL pyridoxine, 0.5 mg/mL nicotinic acid, 100 mg/mL myo-inositol, 2 mg/mL Gly, 30 g/L sucrose, 1 mg/L indolebutyric acid, 0.2 mg/L kinetin, 12 mM AgNO 3 , and 96 mM Na 2 S 2 O 3 .…”

Virus-induced Gene Silencing in Apricot (Prunus armeniaca L.) and Japanese Apricot (P. mume Siebold ^|^amp; Zucc.) with the Apple Latent Spherical Virus Vector System

“…2), a 4.5-kb fragment containing ALSV RNA2 expression cassette tial to silence multicopy genes or genes of different plant species with similar genetic backgrounds (Purkayastha and Dasgupta, 2009). Because of these advantages, several types of viruses have been modified into VIGS vectors and used for the silencing of endogenous genes in herbaceous model plants and crops such as Arabidopsis thaliana, Nicotiana benthamiana, Solanum species, legume species, rice (Oryza sativa L.), wheat (Triticum aestivum L.), barley (Hordeum vulgare L.), maize (Zea mays L.), cotton (Gossypium hirsutum L.), and strawberry (Fragaria × ananassa) (Bennypaul et al, 2012;Brigneti et al, 2004;Chai et al, 2011;Constantin et al, 2004;Ding et al, 2006;Faivre-Rampant et al, 2004;Fu et al, 2005;Holzberg et al, 2002;Turnage et al, 2002;Tuttle et al, 2008;Yan et al, 2012;Zhang and Ghabrial, 2006). In contrast, only a few studies on virus vectors for VIGS in fruit tree species had been reported until recently, mainly because of the lack of efficient viral vectors.…”

Efficient Virus-Induced Gene Silencing in Roots Using a Modified Tobacco Rattle Virus Vector

“…It is believed that VIGS efficiency relies mainly on the capacity of the virus to invade the host and replicate to a sufficient level in targeted tissues (Faivre-Rampant et al, 2004). This suggests that viral tropism would reflect the VIGS response and that it should be stronger in the tissue where viral replication is more favored.…”

“…Indeed, there are numerous examples of endogenous genes that have been successfully silenced in foliar tissues (e.g. phytoene desaturase [pds; Ratcliff et al, 2001] or cellulose synthase [Burton et al, 2000]), flower primordia (FLO/LFY; Ratcliff et al, 2001), meristematic tissue (proliferating cell nuclear antigen; Peele et al, 2001), and potato tubers (pds; Faivre-Rampant et al, 2004). However, root functions have remained an elusive target for many virus-based vectors due to the inefficient invasion of root systems by many viruses.…”

“…These include RNA viruses, such as tobacco mosaic virus (Kumagai et al, 1995;Lacomme et al, 2003), potato virus X (PVX; English et al, 1996;Angell and Baulcombe, 1997;Lu et al, 2003), tobacco rattle virus (TRV; Ratcliff et al, 2001;Liu et al, 2002b), and DNA viruses such as tomato golden mosaic virus (Peele et al, 2001). Recent developments have extended the use of such vectors, originally developed in N. benthamiana, to model plants such as Arabidopsis (Arabidopsis thaliana; Turnage et al, 2002) and crops such as tomato (Lycopersicon esculentum; Liu et al, 2002b), potato (Solanum tuberosum; Faivre-Rampant et al, 2004), and barley (Holzberg et al, 2002;Lacomme et al, 2003). This range of virus-host systems has demonstrated the potential for the functional characterization of endogenous genes in a variety of organs and tissues.…”