Bacterially expressed dsRNA protects maize against SCMV infection

Plant Cell Rep. 2010 Nov;29(11):1261-8. doi: 10.1007/s00299-010-0911-z. Epub 2010 Aug 24.

Abstract

RNA interference (RNAi) is a sequence-specific, posttranscriptional gene silencing (PTGS) process in plants that is mediated by dsRNA homologous to the silenced gene(s). In this study, we report an efficient method to produce dsRNA using a bacterial expression system. Two fragments of the Sugarcane Mosaic Virus (SCMV) CP (coat protein) gene were amplified by RT-PCR, and cloned into the inverted-repeat cloning vector pUCCRNAi. The two recombinant plasmids were transformed individually into E. coli HT115, an RNase-III deficient strain, and dsRNA was induced by isopropyl-β-D: -thiogalactopyranoside (IPTG). The crude extracts of E. coli HT115 containing large amounts of dsRNA were applied to plants as a spray and the experiment confirmed a preventative efficacy. Our findings demonstrated that spraying crude dsRNA-containing extracts inhibited SCMV infection, and the dsRNA derived from an upstream region (CP1) was more effective than was dsRNA derived from a downstream region (CP2) of the SCMV CP gene. The results provide a valuable tool for plant viral control using dsRNA and the PTGS approach.

MeSH terms

  • Capsid Proteins / genetics
  • Escherichia coli / genetics
  • Isopropyl Thiogalactoside / pharmacology
  • Mosaic Viruses / genetics
  • Mosaic Viruses / pathogenicity*
  • Plant Diseases / prevention & control*
  • Plant Diseases / virology
  • Plasmids
  • RNA Interference*
  • RNA, Double-Stranded / pharmacology*
  • Zea mays / virology*

Substances

  • Capsid Proteins
  • RNA, Double-Stranded
  • Isopropyl Thiogalactoside