VIGS – genomics goes functional

Becker, Annette; Lange, Matthias

doi:10.1016/j.tplants.2009.09.002

Cited by 197 publications

(130 citation statements)

References 26 publications

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“…So far, VIGS has been applied to more than 30 plant species (Becker and Lange, 2010), indicating that it has become a powerful tool for functional gene analysis in plants. More than 30 VIGS vectors have been developed so far, including tobacco mosaic virus (Kumagai et al, 1995), potato virus X (Ruiz et al, 1998), tobacco rattle virus (TRV) (Ratcliff et al, 2001), barley stripe mosaic virus (Holzberg et al, 2002), and brome mosaic virus (Ding et al, 2006).…”

Section: Introductionmentioning

confidence: 99%

Optimization of virus-induced gene silencing in pepper (Capsicum annuum L.)

Wang¹,

Li²,

Gong³

et al. 2013

Genet. Mol. Res.

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ABSTRACT. Virus-induced gene silencing is currently a powerful tool for the study of gene function in plants. Here, we optimized the protocol for virus-induced gene silencing, and investigated factors that affect the efficiency of tobacco rattle virus-induced gene silencing in pepper plants. Consequently, an optimal protocol was obtained by the syringe-infiltration method in the leaves of pepper plants. The protocol involves 2-leaf stage plants, preparing the Agrobacterium inoculum at a final OD 600 of 1.0 and then growing the inoculated plants at 22°C. Using this protocol, we achieved high efficiency in silencing CaPDS in different cultivars of pepper plants. We further used the CaPOD gene to illustrate the general reliability of this optimized protocol. Viral symptoms were observed on the leaves of inoculated plants of the Early Calwonder cultivar 25 days post-inoculation, indicating that this protocol can also be used to silence other genes in pepper plants. Real-time polymerase chain reaction analyses revealed that the expression levels of CaPDS and CaPOD were dramatically reduced in inoculated leaves compared to 2493 ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 12 (3): 2492-2506 (2013 Virus-induced gene silencing in pepper control plants. These results demonstrate that the optimized protocol can be applied to functional genomic studies in pepper to investigate genes involved in a wide range of biological processes.

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Section: Introductionmentioning

confidence: 99%

Optimization of virus-induced gene silencing in pepper (Capsicum annuum L.)

Wang¹,

Li²,

Gong³

et al. 2013

Genet. Mol. Res.

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“…In plants, PTGS can be accomplished by creating stable transgene expression hairpin RNA molecules or dsRNA. In the recent decade, virus-induced gene silencing (VIGS), a transient reverse genetics tool for characterizing gene functions, has been widely used in many plants (Kumagai et al, 1995;Burch-Smith et al, 2004;Ding et al, 2006;Becker and Lange, 2010). When a virus infects a plant tissue and spreads systemically throughout the tissue, the endogenous gene transcripts, which are homologous to the fragment inserted into viral vector, are degraded by PTGS.…”

Section: Introductionmentioning

confidence: 99%

“…It can also knockdown either a single copy gene or a gene family, and can be used for studying some genes whose traditional knockout is embryo-lethal or sterile (BurchSmith et al, 2004;Becker and Lange, 2010). Many viruses or virus satellites have been modified into gene silencing vectors.…”

Section: Introductionmentioning

confidence: 99%

Development and application of an efficient virus-induced gene silencing system in Nicotiana tabacum using geminivirus alphasatellite

Huang

Zhang

et al. 2011

J. Zhejiang Univ. Sci. B

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Virus-induced gene silencing (VIGS) is a recently developed technique for characterizing the function of plant genes by gene transcript suppression and is increasingly used to generate transient loss-of-function assays. Here we report that the 2mDNA1, a geminivirus satellite vector, can induce efficient gene silencing in Nicotiana tabacum with Tobacco curly shoot virus. We have successfully silenced the β-glucuronidase (GUS) gene in GUS transgenic N. tabacum plants and the sulphur desaturase (Su) gene in five different N. tabacum cultivars. These pronounced and severe silencing phenotypes are persistent and ubiquitous. Once initiated in seedlings, the silencing phenotype lasted for the entire life span of the plants and silencing could be induced in a variety of tissues and organs including leaf, shoot, stem, root, and flower, and achieved at any growth stage. This system works well between 18-32 °C. We also silenced the NtEDS1 gene and demonstrated that NtEDS1 is essential for N gene mediated resistance against Tobacco mosaic virus in N. tabacum. The above results indicate that this system has great potential as a versatile VIGS system for routine functional analysis of genes in N. tabacum.

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“…Such plant virus-based technology can be applied to facilitate or impede gene expression, resulting in gain-or loss-offunction phenotypes. Although virus-based technology was initially exploited for the purpose of high-level production of foreign proteins, such as recombinant subunit vaccines and pharmaceutical proteins for molecular pharming (Scholthof et al, 1996;Porta and Lomonossoff, 2002), plant RNA and DNA virus-based techniques such as small interfering RNA-mediated virus-induced posttranscriptional gene silencing (VIGS) has been extensively utilized to silence genes for functional genomic studies in dicots and monocots, including plants and crops recalcitrant to classical forward or reverse genetic manipulation (Lindbo et al, 1993;Kumagai et al, 1995;Ruiz et al, 1998;Liu et al, 2002Liu et al, , 2016Becker and Lange, 2010;Senthil-Kumar and Mysore, 2011;Qin et al, 2015). Various virus-based technologies have been developed such as VIGS, microRNA-based VIGS (Tang et al, 2010;Chen et al, 2015aChen et al, , 2015c, virus-based microRNA silencing (Sha et al, 2014), and virus-induced transcriptional gene silencing (Kanazawa et al, 2011;Chen et al, 2015b).…”

mentioning

confidence: 99%

A Virus-Induced Assay for Functional Dissection and Analysis of Monocot and Dicot Flowering Time Genes

Cheng

Chen²,

Shen³

et al. 2017

Plant Physiol.

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Virus-induced flowering (VIF) uses virus vectors to express Flowering Locus T (FT) to induce flowering in plants. This approach has recently attracted wide interest for its practical applications in accelerating breeding in crops and woody fruit trees. However, the insight into VIF and its potential as a powerful tool for dissecting florigenic proteins remained to be elucidated. Here, we describe the mechanism and further applications of Potato virus X (PVX)-based VIF in the short-day Nicotiana tabacum cultivar Maryland Mammoth. Ectopic delivery of Arabidopsis (Arabidopsis thaliana) AtFT by PVX/AtFT did not induce the expression of the endogenous FT ortholog NtFT4; however, it was sufficient to trigger flowering in Maryland Mammoth plants grown under noninductive long-day conditions. Infected tobacco plants developed no systemic symptoms, and the PVX-based VIF did not cause transgenerational flowering. We showed that the PVX-based VIF is a much more rapid method to examine the impacts of single amino acid mutations on AtFT for floral induction than making individual transgenic Arabidopsis lines for each mutation. We also used the PVX-based VIF to demonstrate that adding a His-or FLAG-tag to the N or C terminus of AtFT could affect its florigenic activity and that this system can be applied to assay the function of FT genes from heterologous species, including tomato (Solanum lycopersicum) SFT and rice (Oryza sativa) Hd3a. Thus, the PVX-based VIF represents a simple and efficient system to identify individual amino acids that are essential for FT-mediated floral induction and to test the ability of mono-and dicotyledonous FT genes and FT fusion proteins to induce flowering.Modified plant viruses have emerged as powerful tools for dissecting gene function in plants. Such plant virus-based technology can be applied to facilitate or impede gene expression, resulting in gain-or loss-offunction phenotypes. Although virus-based technology was initially exploited for the purpose of high-level production of foreign proteins, such as recombinant subunit vaccines and pharmaceutical proteins for molecular pharming (Scholthof et al., 1996;Porta and Lomonossoff, 2002), plant RNA and DNA virus-based techniques such as small interfering RNA-mediated virus-induced posttranscriptional gene silencing (VIGS) has been extensively utilized to silence genes for functional genomic studies in dicots and monocots, including plants and crops recalcitrant to classical forward or reverse genetic manipulation (Lindbo et al

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VIGS – genomics goes functional

Cited by 197 publications

References 26 publications

Optimization of virus-induced gene silencing in pepper (Capsicum annuum L.)

Optimization of virus-induced gene silencing in pepper (Capsicum annuum L.)

Development and application of an efficient virus-induced gene silencing system in Nicotiana tabacum using geminivirus alphasatellite

A Virus-Induced Assay for Functional Dissection and Analysis of Monocot and Dicot Flowering Time Genes

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