Efficiency of VIGS and gene expression in a novel bipartite potexvirus vector delivery system as a function of strength of TGB1 silencing suppression

H. S. Lim, A. M. Vaira, L. L. Domier, Sung Chul Lee, Hong Gi Kim, J. Hammond

Research output: Contribution to journalArticlepeer-review

Abstract

We have developed plant virus-based vectors for virus-induced gene silencing (VIGS) and protein expression, based on Alternanthera mosaic virus (AltMV), for infection of a wide range of host plants including Nicotiana benthamiana and Arabidopsis thaliana by either mechanical inoculation of in vitro transcripts or via agroinfiltration. In vivo transcripts produced by co-agroinfiltration of bacteriophage T7 RNA polymerase resulted in T7-driven AltMV infection from a binary vector in the absence of the Cauliflower mosaic virus 35S promoter. An artificial bipartite viral vector delivery system was created by separating the AltMV RNA-dependent RNA polymerase and Triple Gene Block (TGB)123-Coat protein (CP) coding regions into two constructs each bearing the AltMV 5′ and 3′ non-coding regions, which recombined in planta to generate a full-length AltMV genome. Substitution of TGB1 L(88)P, and equivalent changes in other potexvirus TGB1 proteins, affected RNA silencing suppression efficacy and suitability of the vectors from protein expression to VIGS.

Original languageEnglish (US)
Pages (from-to)149-163
Number of pages15
JournalVirology
Volume402
Issue number1
DOIs
StatePublished - Jun 2010

Keywords

  • Agroinfiltration
  • Alternanthera mosaic virus
  • Bipartite launch system
  • In vivo transcription
  • Plant viral vector
  • Potexvirus
  • VIGS

ASJC Scopus subject areas

  • Virology

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