Post-transcriptional Gene Silencing Using Virus-Induced Gene Silencing to Study Plant Gametogenesis in Tomato

Zhengming Wang; Xiao Yu Wang; Claudia Martinho; David C Baulcombe

doi:10.1007/978-1-0716-2253-7_15

Post-transcriptional Gene Silencing Using Virus-Induced Gene Silencing to Study Plant Gametogenesis in Tomato

Zhengming Wang (Lead / Corresponding author), Xiao Yu Wang, Claudia Martinho, David C Baulcombe

Plant Sciences

Research output: Chapter in Book/Report/Conference proceeding › Chapter (peer-reviewed) › peer-review

3 Citations (Scopus)

Abstract

Loss-of-function analyses are essential to dissect the complex nature of biological processes, including gametogenesis. Virus-induced gene silencing (VIGS) has been widely used in crop species as an amenable and rapid way to generate gene knockdowns. As a transient assay, VIGS circumvents the generation of stable transgenic lines through laborious and time-consuming tissue culture techniques. VIGS involves inoculating plants during early development with genetically manipulated viral constructs carrying an endogenous gene target sequence. The viral infection triggers the host plant gene silencing machinery to process the viral genomic RNA into small RNAs (sRNAs) including the gene complementary region. The sRNAs with complementary sequences to the endogenous gene mediate posttranscriptional gene silencing of the targeted gene. Here, we provide a simple and reproducible VIGS protocol employing the tobacco rattle virus (TRV) in tomato (Solanum lycopersicum cv. M82). As it is stable at later developmental stages this approach is suitable for many traits in tomato including gametogenesis and it can be adapted to other crop species.

Original language	English
Title of host publication	Plant Gametogenesis
Subtitle of host publication	Methods and Protocols
Editors	Christophe Lambing
Place of Publication	New York
Publisher	Humana New York, NY
Chapter	15
Pages	201-212
Number of pages	12
Edition	1
ISBN (Electronic)	9781071622537
ISBN (Print)	9781071622520
DOIs	https://doi.org/10.1007/978-1-0716-2253-7_15
Publication status	Published - 2022

Publication series

Name	Methods in Molecular Biology (Clifton, N.J.)
Publisher	Springer Verlag
Volume	2484
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Keywords

Gametogenesis, Plant
Gene Expression Regulation, Plant
Gene Silencing
Genetic Vectors
Solanum lycopersicum/genetics
Plant Viruses/genetics
RNA Interference
RNA, Viral
Nicotiana/genetics

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10.1007/978-1-0716-2253-7_15

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Wang, Z., Wang, X. Y., Martinho, C., & Baulcombe, D. C. (2022). Post-transcriptional Gene Silencing Using Virus-Induced Gene Silencing to Study Plant Gametogenesis in Tomato. In C. Lambing (Ed.), Plant Gametogenesis: Methods and Protocols (1 ed., pp. 201-212). (Methods in Molecular Biology (Clifton, N.J.); Vol. 2484). Humana New York, NY. https://doi.org/10.1007/978-1-0716-2253-7_15

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title = "Post-transcriptional Gene Silencing Using Virus-Induced Gene Silencing to Study Plant Gametogenesis in Tomato",

abstract = "Loss-of-function analyses are essential to dissect the complex nature of biological processes, including gametogenesis. Virus-induced gene silencing (VIGS) has been widely used in crop species as an amenable and rapid way to generate gene knockdowns. As a transient assay, VIGS circumvents the generation of stable transgenic lines through laborious and time-consuming tissue culture techniques. VIGS involves inoculating plants during early development with genetically manipulated viral constructs carrying an endogenous gene target sequence. The viral infection triggers the host plant gene silencing machinery to process the viral genomic RNA into small RNAs (sRNAs) including the gene complementary region. The sRNAs with complementary sequences to the endogenous gene mediate posttranscriptional gene silencing of the targeted gene. Here, we provide a simple and reproducible VIGS protocol employing the tobacco rattle virus (TRV) in tomato (Solanum lycopersicum cv. M82). As it is stable at later developmental stages this approach is suitable for many traits in tomato including gametogenesis and it can be adapted to other crop species.",

keywords = "Gametogenesis, Plant, Gene Expression Regulation, Plant, Gene Silencing, Genetic Vectors, Solanum lycopersicum/genetics, Plant Viruses/genetics, RNA Interference, RNA, Viral, Nicotiana/genetics",

author = "Zhengming Wang and Wang, {Xiao Yu} and Claudia Martinho and Baulcombe, {David C}",

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isbn = "9781071622520",

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Wang, Z, Wang, XY, Martinho, C & Baulcombe, DC 2022, Post-transcriptional Gene Silencing Using Virus-Induced Gene Silencing to Study Plant Gametogenesis in Tomato. in C Lambing (ed.), Plant Gametogenesis: Methods and Protocols. 1 edn, Methods in Molecular Biology (Clifton, N.J.), vol. 2484, Humana New York, NY, New York, pp. 201-212. https://doi.org/10.1007/978-1-0716-2253-7_15

Post-transcriptional Gene Silencing Using Virus-Induced Gene Silencing to Study Plant Gametogenesis in Tomato. / Wang, Zhengming (Lead / Corresponding author); Wang, Xiao Yu; Martinho, Claudia et al.
Plant Gametogenesis: Methods and Protocols. ed. / Christophe Lambing. 1. ed. New York: Humana New York, NY, 2022. p. 201-212 (Methods in Molecular Biology (Clifton, N.J.); Vol. 2484).

Research output: Chapter in Book/Report/Conference proceeding › Chapter (peer-reviewed) › peer-review

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PY - 2022

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N2 - Loss-of-function analyses are essential to dissect the complex nature of biological processes, including gametogenesis. Virus-induced gene silencing (VIGS) has been widely used in crop species as an amenable and rapid way to generate gene knockdowns. As a transient assay, VIGS circumvents the generation of stable transgenic lines through laborious and time-consuming tissue culture techniques. VIGS involves inoculating plants during early development with genetically manipulated viral constructs carrying an endogenous gene target sequence. The viral infection triggers the host plant gene silencing machinery to process the viral genomic RNA into small RNAs (sRNAs) including the gene complementary region. The sRNAs with complementary sequences to the endogenous gene mediate posttranscriptional gene silencing of the targeted gene. Here, we provide a simple and reproducible VIGS protocol employing the tobacco rattle virus (TRV) in tomato (Solanum lycopersicum cv. M82). As it is stable at later developmental stages this approach is suitable for many traits in tomato including gametogenesis and it can be adapted to other crop species.

AB - Loss-of-function analyses are essential to dissect the complex nature of biological processes, including gametogenesis. Virus-induced gene silencing (VIGS) has been widely used in crop species as an amenable and rapid way to generate gene knockdowns. As a transient assay, VIGS circumvents the generation of stable transgenic lines through laborious and time-consuming tissue culture techniques. VIGS involves inoculating plants during early development with genetically manipulated viral constructs carrying an endogenous gene target sequence. The viral infection triggers the host plant gene silencing machinery to process the viral genomic RNA into small RNAs (sRNAs) including the gene complementary region. The sRNAs with complementary sequences to the endogenous gene mediate posttranscriptional gene silencing of the targeted gene. Here, we provide a simple and reproducible VIGS protocol employing the tobacco rattle virus (TRV) in tomato (Solanum lycopersicum cv. M82). As it is stable at later developmental stages this approach is suitable for many traits in tomato including gametogenesis and it can be adapted to other crop species.

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KW - Gene Expression Regulation, Plant

KW - Gene Silencing

KW - Genetic Vectors

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KW - Plant Viruses/genetics

KW - RNA Interference

KW - RNA, Viral

KW - Nicotiana/genetics

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M3 - Chapter (peer-reviewed)

C2 - 35461454

SN - 9781071622520

T3 - Methods in Molecular Biology (Clifton, N.J.)

SP - 201

EP - 212

BT - Plant Gametogenesis

A2 - Lambing, Christophe

PB - Humana New York, NY

CY - New York

ER -

Wang Z, Wang XY, Martinho C, Baulcombe DC. Post-transcriptional Gene Silencing Using Virus-Induced Gene Silencing to Study Plant Gametogenesis in Tomato. In Lambing C, editor, Plant Gametogenesis: Methods and Protocols. 1 ed. New York: Humana New York, NY. 2022. p. 201-212. (Methods in Molecular Biology (Clifton, N.J.)). Epub 2022 Apr 24. doi: 10.1007/978-1-0716-2253-7_15

Post-transcriptional Gene Silencing Using Virus-Induced Gene Silencing to Study Plant Gametogenesis in Tomato

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