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

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

2 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 languageEnglish
Title of host publicationPlant Gametogenesis
Subtitle of host publicationMethods and Protocols
EditorsChristophe Lambing
Place of PublicationNew York
PublisherHumana New York, NY
Chapter15
Pages201-212
Number of pages12
Edition1
ISBN (Electronic)9781071622537
ISBN (Print)9781071622520
DOIs
Publication statusPublished - 2022

Publication series

NameMethods in Molecular Biology (Clifton, N.J.)
PublisherSpringer Verlag
Volume2484
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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