title: The Arabidopsis NADPH oxidases RbohD and RbohF display differential expression patterns and contributions during plant immunity.
abstract: Plant NADPH oxidases, also known as respiratory burst oxidase homologues (RBOHs), produce reactive oxygen species (ROS) that perform a wide range of functions. RbohD and RbohF, two of the 10 Rboh genes present in Arabidopsis, are pleiotropic and mediate diverse physiological processes including the response to pathogens. We hypothesized that the spatio-temporal control of RbohD and RbohF gene expression might be critical in determining their multiplicity of functions. Transgenic Arabidopsis plants with RbohD and RbohF promoter fusions to β-glucuronidase and Luciferase reporter genes were generated. Analysis of these plants revealed a differential expression pattern for RbohD and RbohF throughout plant development and during immune responses. RbohD and RbohF gene expression was differentially modulated by pathogen-associated molecular patterns. Histochemical stains and in vivo expression analysis showed a correlation between the level of RbohD and RbohF promoter activity, H2O2 accumulation and the amount of cell death in response to the pathogenic bacterium Pseudomonas syringae pv. tomato DC3000 and the necrotrophic fungus Plectosphaerella cucumerina. A promoter-swap strategy revealed that the promoter region of RbohD was required to drive production of ROS by this gene in response to pathogens. Moreover, RbohD promoter was activated during Arabidopsis interaction with a non-virulent P. cucumerina isolate, and susceptibility tests with the double mutant rbohD rbohF uncovered a new function for these oxidases in basal resistance. Altogether, our results suggest that differential spatio-temporal expression of the Rboh genes contributes to fine-tune RBOH/NADPH oxidase-dependent ROS production and signaling in Arabidopsis immunity.
authors.affiliation: Centro de Biotecnología y Genómica de Plantas (UPM, INIA), Escuela Superior Técnica de Ingenieros Agrónomos, Universidad Politécnica de Madrid, Campus Montegancedo, Autopista M40 Km 38, Pozuelo de Alarcón, 28223, Madrid, Spain miguelangel.torres@upm.es.
authors.forename: Jorge
authors.initials: J
authors.lastname: Morales
authors.affiliation: The Sainsbury Laboratory, Norwich Research Park, Norwich NR4 7UH, UK RIKEN Center for Sustainable Resource Science, Plant Immunity Research Group, Suehiro-cho 1-7-22 Tsurumi-ku, Yokohama 230-0045, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Kadota
authors.affiliation: The Sainsbury Laboratory, Norwich Research Park, Norwich NR4 7UH, UK.
authors.forename: Cyril
authors.initials: C
authors.lastname: Zipfel
authors.affiliation: Centro de Biotecnología y Genómica de Plantas (UPM, INIA), Escuela Superior Técnica de Ingenieros Agrónomos, Universidad Politécnica de Madrid, Campus Montegancedo, Autopista M40 Km 38, Pozuelo de Alarcón, 28223, Madrid, Spain.
authors.forename: Antonio
authors.initials: A
authors.lastname: Molina
authors.affiliation: Centro de Biotecnología y Genómica de Plantas (UPM, INIA), Escuela Superior Técnica de Ingenieros Agrónomos, Universidad Politécnica de Madrid, Campus Montegancedo, Autopista M40 Km 38, Pozuelo de Alarcón, 28223, Madrid, Spain miguelangel.torres@upm.es.
authors.forename: Miguel-Angel
authors.initials: MA
authors.lastname: Torres
mesh_terms: D017360:Arabidopsis / Q000196:embryology / Q000201:enzymology* / Q000235:genetics* / Q000276:immunology; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D001203:Ascomycota / Q000502:physiology; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D017930:Genes, Reporter; D005816:Genetic Complementation Test; D009154:Mutation / Q000235:genetics; D019255:NADPH Oxidases / Q000235:genetics / Q000378:metabolism*; D000069452:Pathogen-Associated Molecular Pattern Molecules / Q000378:metabolism; D057865:Plant Immunity / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D044224:Pseudomonas syringae / Q000502:physiology; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; D019255:NADPH Oxidases; D000069452:Pathogen-Associated Molecular Pattern Molecules; D017382:Reactive Oxygen Species; C568857:AT5G47910 protein, Arabidopsis; C000589475:RBOH F protein, Arabidopsis
keywords: Arabidopsis; NADPH oxidase; RbohD; RbohF, reactive oxygen species.; pathogen