R-type anion channel activation is an essential step for ROS-dependent innate immune response in Arabidopsis suspension cells
Jean Colcombet B D , Yves Mathieu A D , Remi Peyronnet A , Nicolas Agier C , Françoise Lelièvre A , Hélène Barbier-Brygoo A and Jean-Marie Frachisse A EA Institut des Sciences du Végétal, CNRS UPR 2355, 22 Avenue de la Terrasse, 91198 Gif sur Yvette, France.
B Present address: Unité de Recherche en Génomique Végétale, 2 rue Gaston Crémieux, 91057 Evry, France.
C Present address: CNRS-CGM, 14 Avenue de la Terrasse, 91198 Gif sur Yvette, France.
D These authors contributed equally to the work.
E Corresponding author. Email: frachisse@isv.cnrs-gif.fr
Functional Plant Biology 36(9) 832-843 https://doi.org/10.1071/FP09096
Submitted: 1 May 2009 Accepted: 23 July 2009 Published: 3 September 2009
Abstract
Plants are constantly exposed to environmental biotic and abiotic stresses. Plants cells perceive these factors and trigger early responses followed by delayed and complex adaptation processes. Using cell suspensions of Arabidopsis thaliana (L.) as a cellular model, we investigated the role of plasma membrane anion channels in Reactive Oxygen Species (ROS) production and in cell death which occurs during non-host pathogen infection. Protoplasts derived from Arabidopsis suspension cells display two anion currents with characteristics very similar to those of the slow nitrate-permeable (S-type) and rapid sulfate-permeable (R-type) channels previously characterised in hypocotyl cells and other cell types. Using seven inhibitors, we showed that the R-type channel and ROS formation in cell cultures present similar pharmacological profiles. The efficiency of anion channel blockers to inhibit ROS production was independent of the nature of the triggering signal (osmotic stress or general elicitors of plant defence), indicating that the R-type channel represents a crossroad in the signalling pathways leading to ROS production. In a second step, we show that treatment with R-type channel blockers accelerates cell death triggered by the non-specific plant pathogen Xanthomonas campestris. Finally, we discuss the hypothesis that the R-type channel is involved in innate immune response allowing cell defence via antibacterial ROS production.
Additional keywords: activated oxygen species, AOS, Arabidopsis thaliana, cell suspensions, defence, immunity, innate immunity, PAMPs, pathogen-associated molecular patterns, pharmacology, plasma membrane, Reactive Oxygen Species, sulphate, transduction.
Acknowledgements
The authors thank their colleagues, S. Thomine, J. Guern, C. Lauriere, F. Bouteau and E. Diatloff for their advice and A. Kiwi Winger for MS reading. We thank Thomas Boller and Georg Felix (Botanisches Institut der Universitat Basel, Switzerland) for the kind gift of flagellin 22. Dominique Roby (Laboratoire des Interactions Plantes Microorganismes, CNRS/INRA, France) is acknowledged for providing Xanthomonas campestris strain. We thank Hoffmann-La Roche SA (Roche, Basel, Switzerland) for providing us with mibefradil. This work was funded by the French Centre National de la Recherche Scientifique and by a European grant from a Research Training Network (NICIP CT-2002–000245).
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