title: A simple agroinfiltration method for transient gene expression in plant leaf discs.
abstract: In the present study, we developed a simple transient gene expression system based on Agrobacterium-mediated transformation. Vacuum infiltration was applied to leaf discs from Nicotiana benthamiana plants with Agrobacterium suspension solution under conventional vacuum conditions in a needleless plastic syringe. Model proteins, green fluorescent protein, β-glucuronidase, mouse granulocyte-macrophage colony-stimulating factor, and human fibroblast growth factor 1 were successfully expressed in leaf discs within 4 days after infiltration. In addition, the functional evaluation of viral RNA silencing suppressors, Artichoke mottled crinkle virus p19 protein, was also performed. Using this method, the contamination and diffusion of genetically modified bacterium to the environment and important transgenic plants were prevented. This method can be conducted without specialized apparatuses or large amounts of Agrobacterium suspension solutions; thus, the simultaneous evaluation of multiple vectors will be easily possible.
authors.affiliation: Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 2-17-2-1 Tsukisamu-Higashi, Toyohira-ku, Sapporo 062-8517, Japan. Electronic address: matsuo-kouki@aist.go.jp.
authors.forename: Kouki
authors.initials: K
authors.lastname: Matsuo
authors.affiliation: Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 2-17-2-1 Tsukisamu-Higashi, Toyohira-ku, Sapporo 062-8517, Japan.
authors.forename: Noriho
authors.initials: N
authors.lastname: Fukuzawa
authors.affiliation: Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 2-17-2-1 Tsukisamu-Higashi, Toyohira-ku, Sapporo 062-8517, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Matsumura
mesh_terms: D060054:Agrobacterium / Q000235:genetics; D000818:Animals; D016220:Fibroblast Growth Factor 1 / Q000235:genetics; D018506:Gene Expression Regulation, Plant / Q000235:genetics*; D005966:Glucuronidase / Q000235:genetics; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D051379:Mice; D018515:Plant Leaves / Q000235:genetics* / Q000382:microbiology; D010942:Plant Viruses / Q000235:genetics; D030821:Plants, Genetically Modified / Q000235:genetics / Q000382:microbiology; D034622:RNA Interference; D014026:Nicotiana / Q000235:genetics / Q000382:microbiology; D014618:Vacuum; D014764:Viral Proteins / Q000235:genetics
chemical_list: D014764:Viral Proteins; D016220:Fibroblast Growth Factor 1; D049452:Green Fluorescent Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D005966:Glucuronidase
keywords: Agroinfiltration; Leaf disc; Nicotiana benthamiana; Transient gene expression; Vacuum infiltration