Plant Viruses as Virus Induced Gene Silencing (VIGS) Vectors

2019 ◽  
pp. 517-526
Author(s):  
Sunny Dhir ◽  
Ashish Srivastava ◽  
Nobiyuki Yoshikawa ◽  
S. M. Paul Khurana
Keyword(s):  
Gene Silencing ◽  
Plant Viruses ◽  
Virus Induced Gene Silencing

scholarly journals Efficient Infection of Nicotiana benthamiana by Tomato bushy stunt virus Is Facilitated by the Coat Protein and Maintained by p19 Through Suppression of Gene Silencing

2002 ◽  
Vol 15 (3) ◽  
pp. 193-202 ◽  
Author(s):  
Feng Qu ◽  
T. Jack Morris
Keyword(s):  
Coat Protein ◽  
Gene Silencing ◽  
Vascular System ◽  
Systemic Infection ◽  
Red Clover ◽  
Plant Viruses ◽  
Tomato Bushy Stunt Virus ◽  
Careful Examination ◽  
Virus Induced Gene Silencing ◽  
Systemic Spread

Tomato bushy stunt virus (TBSV) is one of few RNA plant viruses capable of moving systemically in some hosts in the absence of coat protein (CP). TBSV also encodes another protein (p19) that is not required for systemic movement but functions as a symptom determinant in Nicotiana ben-thamiana. Here, the role of both CP and p19 in the systemic spread has been reevaluated by utilizing transgenic N. ben-thamiana plants expressing the movement protein (MP) of Red clover necrotic mosaic virus and chimeric TBSV mutants that express CP of Turnip crinkle virus. Through careful examination of the infection phenotype of a series of mutants with changes in the CP and p19 genes, we demonstrate that both of these genes are required for efficient systemic invasion of TBSV in N. benthamiana. The CP likely enables efficient viral unloading from the vascular system in the form of assembled virions, whereas p19 enhances systemic infection by suppressing the virus-induced gene silencing.

VIGS technology: an attractive tool for functional genomics studies in legumes

2013 ◽  
Vol 40 (12) ◽  
pp. 1234 ◽  
Author(s):  
Stéphanie Pflieger ◽  
Manon M. S. Richard ◽  
Sophie Blanchet ◽  
Chouaib Meziadi ◽  
Valérie Geffroy
Keyword(s):  
Gene Silencing ◽  
Functional Genomics ◽  
Target Gene ◽  
Reverse Genetics ◽  
Plant Viruses ◽  
Transcriptional Gene Silencing ◽  
Full Potential ◽  
Legume Species ◽  
Virus Induced Gene Silencing ◽  
Apple Latent Spherical Virus

Legume species are among the most important crops worldwide. In recent years, six legume genomes have been completely sequenced, and there is now an urgent need for reverse-genetics tools to validate genes affecting yield and product quality. As most legumes are recalcitrant to stable genetic transformation, virus-induced gene silencing (VIGS) appears to be a powerful alternative technology for determining the function of unknown genes. VIGS technology is based on the property of plant viruses to trigger a defence mechanism related to post-transcriptional gene silencing (PTGS). Infection by a recombinant virus carrying a fragment of a plant target gene will induce homology-dependent silencing of the endogenous target gene. Several VIGS systems have been developed for legume species since 2004, including those based on Bean pod mottle virus, Pea early browning virus, and Apple latent spherical virus, and used in reverse-genetics studies of a wide variety of plant biological processes. In this work, we give an overview of the VIGS systems available for legumes, and present their successful applications in functional genomics studies. We also discuss the limitations of these VIGS systems and the future challenges to be faced in order to use VIGS to its full potential in legume species.

Virus-induced Gene Silencing (VIGS) in Barley Seedling Leaves

BIO-PROTOCOL ◽  
2015 ◽  
Vol 5 (12) ◽  
Author(s):  
Lokanadha Gunupuru ◽  
Shahin Ali ◽  
Fiona Doohan ◽  
Steven Scofield
Keyword(s):  
Gene Silencing ◽  
Virus Induced Gene Silencing ◽  
Barley Seedling

scholarly journals Cucumber mosaic virus-induced gene silencing in banana

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Yuh Tzean ◽  
Ming-Chi Lee ◽  
Hsiao-Hsuan Jan ◽  
Yi-Shu Chiu ◽  
Tsui-Chin Tu ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Virus Induced Gene Silencing

scholarly journals A cassava common mosaic virus vector for virus-induced gene silencing in cassava

Plant Methods ◽  
2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Decai Tuo ◽  
Peng Zhou ◽  
Pu Yan ◽  
Hongguang Cui ◽  
Yang Liu ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Mosaic Virus ◽  
Gene Function ◽  
Viral Vector ◽  
Function Analysis ◽  
Systemic Infection ◽  
Cdna Clones ◽  
Virus Induced Gene Silencing ◽  
Efficient Gene ◽  
Gene Function Analysis

Abstract Background Cassava is an important crop for food security and industry in the least-developed and developing countries. The completion of the cassava genome sequence and identification of large numbers of candidate genes by next-generation sequencing provide extensive resources for cassava molecular breeding and increase the need for rapid and efficient gene function analysis systems in cassava. Several plant virus-induced gene silencing (VIGS) systems have been developed as reverse genetic tools for rapid gene function analysis in cassava. However, these VIGS vectors could cause severe viral symptoms or inefficient gene silencing. Results In this study, we constructed agroinfection-compatible infectious cDNA clones of cassava common mosaic virus isolate CM (CsCMV-CM, genus Potexvirus, family Alphaflexiviridae) that causes systemic infection with mild symptoms in cassava. CsCMV-CM was then modified to a viral vector carrying the Nimble cloning frame, which facilitates the rapid and high-throughput cloning of silencing fragments into the viral genome. The CsCMV-based vector successfully silenced phytoene desaturase (PDS) and magnesium chelatase subunit I (ChlI) in different cassava varieties and Nicotiana benthamiana. The silencing of the ChlI gene could persist for more than two months. Conclusions This CsCMV-based VIGS system provides a new tool for rapid and efficient gene function studies in cassava.

scholarly journals Virus-induced gene silencing as a tool for analysis of gene functions in plants

Uirusu ◽  
2010 ◽  
Vol 60 (2) ◽  
pp. 155-162 ◽  
Author(s):  
Noriko YAMAGISHI ◽  
Nobuyuki YOSHIKAWA
Keyword(s):  
Gene Silencing ◽  
Virus Induced Gene Silencing ◽  
Gene Functions

scholarly journals Application of virus-induced gene silencing for identification of FHB resistant genes

2019 ◽  
Vol 18 (10) ◽  
pp. 2183-2192 ◽  
Author(s):  
Yan-hui FAN ◽  
Bing-qian HOU ◽  
Pei-sen SU ◽  
Hong-yan WU ◽  
Gui-ping WANG ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Virus Induced Gene Silencing ◽  
Resistant Genes

scholarly journals Simultaneous silencing of two different Arabidopsis genes with a novel virus-induced gene silencing vector

Plant Methods ◽  
2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Kunxin Wu ◽  
Yadan Wu ◽  
Chunwei Zhang ◽  
Yan Fu ◽  
Zhixin Liu ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Double Mutant ◽  
Target Genes ◽  
Rna Binding ◽  
Protein Product ◽  
Antiviral Defense ◽  
Virus Induced Gene Silencing ◽  
Argonaute 2 ◽  
Plant Genes ◽  
Silencing Pathways

Abstract Background Virus-induced gene silencing (VIGS) is a useful tool for functional characterizations of plant genes. However, the penetrance of VIGS varies depending on the genes to be silenced, and has to be evaluated by examining the transcript levels of target genes. Results In this report, we report the development of a novel VIGS vector that permits a preliminary assessment of the silencing penetrance. This new vector is based on an attenuated variant of Turnip crinkle virus (TCV) known as CPB that can be readily used in Arabidopsis thaliana to interrogate genes of this model plant. A CPB derivative, designated CPB1B, was produced by inserting a 46 nucleotide section of the Arabidopsis PHYTOENE DESATURASE (PDS) gene into CPB, in antisense orientation. CPB1B induced robust PDS silencing, causing easily visible photobleaching in systemically infected Arabidopsis leaves. More importantly, CPB1B can accommodate additional inserts, derived from other Arabidopsis genes, causing the silencing of two or more genes simultaneously. With photobleaching as a visual marker, we adopted the CPB1B vector to validate the involvement of DICER-LIKE 4 (DCL4) in antiviral defense against TCV. We further revealed the involvement of ARGONAUTE 2 (AGO2) in PDS silencing and antiviral defense against TCV in dcl2drb4 double mutant plants. These results demonstrated that DOUBLE-STRANDED RNA-BINDING PROTEIN 4 (DRB4), whose protein product (DRB4) commonly partners with DCL4 in the antiviral silencing pathway, was dispensable for PDS silencing induced by CPB1B derivative in dcl2drb4 double mutant plants. Conclusions The CPB1B-based vector developed in this work is a valuable tool with visualizable indicator of the silencing penetrance for interrogating Arabidopsis genes, especially those involved in the RNA silencing pathways.

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