title: An efficient Agrobacterium-mediated transient transformation of Arabidopsis.
abstract: Agrobacterium tumefaciens-mediated transient transformation has been a useful procedure for characterization of proteins and their functions in plants, including analysis of protein-protein interactions. Agrobacterium-mediated transient transformation of Nicotiana benthamiana by leaf infiltration has been widely used due to its ease and high efficiency. However, in Arabidopsis this procedure has been challenging. Previous studies suggested that this difficulty was caused by plant immune responses triggered by perception of Agrobacterium. Here, we report a simple and robust method for Agrobacterium-mediated transient transformation in Arabidopsis. AvrPto is an effector protein from the bacterial plant pathogen Pseudomonas syringae that suppresses plant immunity by interfering with plant immune receptors. We used transgenic Arabidopsis plants that conditionally express AvrPto under the control of a dexamethasone (DEX)-inducible promoter. When the transgenic plants were pretreated with DEX prior to infection with Agrobacterium carrying a β-glucuronidase (GUS, uidA) gene with an artificial intron and driven by the CaMV 35S promoter, transient GUS expression was dramatically enhanced compared to that in mock-pretreated plants. This transient expression system was successfully applied to analysis of the subcellular localization of a cyan fluorescent protein (CFP) fusion and a protein-protein interaction in Arabidopsis. Our findings enable efficient use of Agrobacterium-mediated transient transformation in Arabidopsis thaliana.
authors: Department of Plant Biology, Microbial and Plant Genomics Institute, University of Minnesota, St Paul, MN 55108, USA.
authors: Kenichi
authors: K
authors: Tsuda
authors: Yiping
authors: Y
authors: Qi
authors: Le V
authors: le V
authors: Nguyen
authors: Gerit
authors: G
authors: Bethke
authors: Yayoi
authors: Y
authors: Tsuda
authors: Jane
authors: J
authors: Glazebrook
authors: Fumiaki
authors: F
authors: Katagiri
mesh_terms: D016960:Agrobacterium tumefaciens / Q000502:physiology*; D017360:Arabidopsis / Q000187:drug effects / Q000235:genetics* / Q000382:microbiology / Q000502:physiology; D001426:Bacterial Proteins / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics; D003907:Dexamethasone / Q000494:pharmacology; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins; D018515:Plant Leaves / Q000187:drug effects / Q000235:genetics / Q000382:microbiology / Q000502:physiology; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic / Q000235:genetics; D025941:Protein Interaction Mapping; D044224:Pseudomonas syringae / Q000235:genetics; D014170:Transformation, Genetic / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; C485182:Cyan Fluorescent Protein; D004269:DNA, Bacterial; C081695:avrPto protein, Pseudomonas syringae; D049452:Green Fluorescent Proteins; D003907:Dexamethasone; D005966:Glucuronidase