title: An Arabidopsis and tomato mesophyll protoplast system for fast identification of early MAMP-triggered immunity-suppressing effectors.
abstract: Transient expression in plant mesophyll protoplasts allows rapid characterisation of gene functions in vivo in a simplified and synchronized manner without bias due to the use of bacteria-based gene or protein delivery systems. It offers the possibility to test whether microbial effectors can subvert early events of plant immune signaling that are activated upon recognition of Microbe-Associated Molecular Patterns (MAMPs), the so-called MAMP-triggered immunity (MTI). Here, we describe the isolation and transfection with effector genes of Arabidopsis thaliana and Solanum lycopersicum mesophyll protoplasts, the use of a non-invasive luciferase reporter assay and a simple method to detect activated Mitogen-Activated Protein Kinases (MAPKs) to identify and study, in a medium-throughput manner, new effectors suppressing early signal transduction events of MTI.
authors.affiliation: Department of Plant Biochemistry, Centre for Plant Molecular Biology, Eberhard Karls University, Auf der Morgenstelle 32, 72076, Tübingen, Germany.
authors.forename: Malou
authors.initials: M
authors.lastname: Fraiture
authors.forename: Xiangzi
authors.initials: X
authors.lastname: Zheng
authors.forename: Frédéric
authors.initials: F
authors.lastname: Brunner
mesh_terms: D017360:Arabidopsis / Q000201:enzymology / Q000254:growth & development / Q000276:immunology*; D001426:Bacterial Proteins / Q000378:metabolism*; D001681:Biological Assay / Q000379:methods*; D016923:Cell Death; D004789:Enzyme Activation; D057075:Enzyme Assays; D017930:Genes, Reporter; D005966:Glucuronidase / Q000378:metabolism; D015151:Immunoblotting; D008156:Luciferases / Q000378:metabolism; D018551:Solanum lycopersicum / Q000254:growth & development / Q000276:immunology*; D058503:Mesophyll Cells / Q000166:cytology*; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D057865:Plant Immunity; D011092:Polyethylene Glycols / Q000737:chemistry; D011523:Protoplasts / Q000166:cytology / Q000378:metabolism*; D051192:Receptors, Pattern Recognition / Q000378:metabolism*; D014162:Transfection
chemical_list: D001426:Bacterial Proteins; D051192:Receptors, Pattern Recognition; D011092:Polyethylene Glycols; D008156:Luciferases; D020928:Mitogen-Activated Protein Kinases; D005966:Glucuronidase