Virus-Induced Gene Silencing in Poaceae Using a Foxtail Mosaic Virus Vector

Methods Mol Biol. 2020:2172:15-25. doi: 10.1007/978-1-0716-0751-0_2.

Abstract

Virus-induced gene silencing (VIGS) is a powerful tool for rapidly knocking down the expression of plant genes to elucidate functional genomics. We have established a VIGS vector for monocot plants derived from Foxtail mosaic virus (FoMV), a positive-sense single-stranded RNA virus. For silencing a targeted gene, plant gene fragment was inserted into the vector between open reading frame 4 (ORF4) and ORF5 under the control of a duplicated coat protein promoter. Plants of different monocot species were infected by mechanical inoculation with sap from FoMV derivative-infected Chenopodium quinoa leaves. Gene silencing was typically observed within 2-3 weeks after inoculation. In this chapter, we describe the detailed protocol for silencing a target gene in various Poaceae plants by using FoMV-based vectors.

Keywords: FoMV-based VIGS vector; Foxtail mosaic virus (FoMV); Monocot; Poaceae; Posttranscriptional gene silencing (PTGS); Virus-induced gene silencing (VIGS).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Gene Expression Regulation, Plant / genetics
  • Gene Expression Regulation, Plant / physiology
  • Gene Silencing / physiology
  • Open Reading Frames / genetics*
  • Plant Proteins / genetics
  • Plant Proteins / metabolism*
  • Plants, Genetically Modified / genetics
  • Plants, Genetically Modified / metabolism*
  • Poaceae / genetics*
  • Poaceae / virology*
  • Potexvirus / genetics*

Substances

  • Plant Proteins

Supplementary concepts

  • Foxtail mosaic virus