title: Tombusvirus-based vector systems to permit over-expression of genes or that serve as sensors of antiviral RNA silencing in plants.
abstract: A next generation Tomato bushy stunt virus (TBSV) coat protein gene replacement vector system is described that can be applied by either RNA inoculation or through agroinfiltration. A vector expressing GFP rapidly yields high levels of transient gene expression in inoculated leaves of various plant species, as illustrated for Nicotiana benthamiana, cowpea, tomato, pepper, and lettuce. A start-codon mutation to down-regulate the dose of the P19 silencing suppressor reduces GFP accumulation, whereas mutations that result in undetectable levels of P19 trigger rapid silencing of GFP. Compared to existing virus vectors the TBSV system has a unique combination of a very broad host range, rapid and high levels of replication and gene expression, and the ability to regulate its suppressor. These features are attractive for quick transient assays in numerous plant species for over-expression of genes of interest, or as a sensor to monitor the efficacy of antiviral RNA silencing.
authors.affiliation: Department of Plant Pathology & Microbiology, Texas A&M University, College Station, TX 77843, USA.
authors.forename: Malika
authors.initials: M
authors.lastname: Shamekova
authors.affiliation: Department of Plant Pathology & Microbiology, Texas A&M University, College Station, TX 77843, USA.
authors.forename: Maria R
authors.initials: MR
authors.lastname: Mendoza
authors.affiliation: Department of Plant Pathology & Microbiology, Texas A&M University, College Station, TX 77843, USA.
authors.forename: Yi-Cheng
authors.initials: YC
authors.lastname: Hsieh
authors.affiliation: Department of Plant Pathology, Ohio State University, Wooster, OH 44691, USA.
authors.forename: John
authors.initials: J
authors.lastname: Lindbo
authors.affiliation: Department of Plant Pathology & Microbiology, Texas A&M University, College Station, TX 77843, USA.
authors.forename: Rustem T
authors.initials: RT
authors.lastname: Omarov
authors.affiliation: Department of Plant Pathology & Microbiology, Texas A&M University, College Station, TX 77843, USA. Electronic address: herscho@tamu.edu.
authors.forename: Herman B
authors.initials: HB
authors.lastname: Scholthof
mesh_terms: D015870:Gene Expression; D020868:Gene Silencing; D018014:Gene Transfer Techniques / Q000295:instrumentation; D005822:Genetic Vectors / Q000235:genetics* / Q000502:physiology; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics / Q000276:immunology; D010944:Plants / Q000235:genetics* / Q000276:immunology / Q000821:virology*; D030821:Plants, Genetically Modified / Q000235:genetics / Q000276:immunology / Q000821:virology; D017864:Tombusvirus / Q000235:genetics* / Q000502:physiology; D019076:Transgenes
chemical_list: D010940:Plant Proteins; D049452:Green Fluorescent Proteins
keywords: Antiviral RNA silencing; Gene vector; Suppression; TBSV; Virus