Efficiency of VIGS and gene expression in a novel bipartite potexvirus vector delivery system as a function of strength of TGB1 silencing suppression.
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.
Lim
Hyoun-Sub
HS
USDA-ARS, Plant Sciences Institute, Molecular Plant Pathology Laboratory, 10300 Baltimore Avenue, Beltsville, MD 20705, USA. Hyoun-Sub.Lim@ars.usda.gov
Vaira
Anna Maria
AM
Domier
Leslie L
LL
Lee
Sung Chul
SC
Kim
Hong Gi
HG
Hammond
John
J
D019943:Amino Acid Substitution / Q000235:genetics; D017360:Arabidopsis / Q000821:virology*; D017123:Bacteriophage T7 / Q000235:genetics; D017796:Caulimovirus / Q000235:genetics; D015870:Gene Expression; D020868:Gene Silencing; D005814:Genes, Viral; D005822:Genetic Vectors; D020125:Mutation, Missense; D017863:Potexvirus / Q000235:genetics* / Q000276:immunology; D011401:Promoter Regions, Genetic; D016601:RNA-Binding Proteins / Q000235:genetics*; D014026:Nicotiana / Q000821:virology*
D016601:RNA-Binding Proteins