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Plant and Cell Physiology Advance Access originally published online on August 29, 2008
Plant and Cell Physiology 2008 49(10):1495-1507; doi:10.1093/pcp/pcn126
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© The Author 2008. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Anion Channel Activation and Proton Pumping Inhibition Involved in the Plasma Membrane Depolarization Induced by ABA in Arabidopsis thaliana Suspension Cells are Both ROS Dependent

Jacques Trouverie1,4, Guillaume Vidal1, Zongshen Zhang1,5, Caroline Sirichandra1,6, Karine Madiona1, Zahia Amiar1, Jean-Louis Prioul2, Emmanuelle Jeannette3, Jean-Pierre Rona1,* and Mathias Brault1,6

1 Laboratoire d’Electrophysiologie des Membranes, EA 3514, Université Paris-Diderot, 2 place Jussieu, 75251 Paris cedex 05, France
2 Institut de Biotechnologie des Plantes, Université Paris-Sud, Bâtiment 630, 91405 Orsay cedex, France
3 Physiologie Cellulaire et Moléculaire des Plantes, Université Pierre et Marie Curie, UMR CNRS 7180, Le Raphaël, 3 rue Galilée, 94200, Ivry-sur-Seine, France

*Corresponding author: E-mail, jean-pierre.rona{at}univ-paris-diderot.fr; Fax, +33-1-44-27-78-13.


  Abstract

In Arabidopsis thaliana suspension cells, ABA was previously shown to promote the activation of anion channels and the reduction of proton pumping that both contribute to the plasma membrane depolarization. These two ABA responses were shown to induce two successive [Ca2+]cyt spikes. As reactive oxygen species (ROS) have emerged as components of ABA signaling pathways especially by promoting [Ca2+]cyt variations, we studied whether ROS were involved in the regulation of anion channels and proton pumps activities. Here we demonstrated that ABA induced ROS production which triggered the second of the two [Ca2+]cyt increases observed in response to ABA. Blocking ROS generation using diphenyleneiodonium (DPI) impaired the proton pumping reduction, the anion channel activation and the RD29A gene expression in response to ABA. Furthermore, H2O2 was shown to activate anion channels and to inhibit plasma membrane proton pumping, as did ABA. However, ROS partially mimicked ABA's effects since H2O2 treatment elicited anion channel activation but not the subsequent expression of the RD29A gene as did ABA. This suggests that expression of the RD29A gene in response to ABA results from the activation of multiple concomitant signaling pathways: blocking of one of them would impair gene expression whereas stimulating only one would not. We conclude that ROS are a central messenger of ABA in the signaling pathways leading to the plasma membrane depolarization induced by ABA.

Keywords: ABA - Anion channel - Arabidopsis thaliana - Calcium - Proton pump - ROS

Abbreviations: 9-AC, 9-anthracene carboxylic acid; BL, blue light; CBL, calcineurin B-like calcium sensor; CDPK, calcium-dependent protein kinase; DPI, diphenyleneiodonium; HACC, hyperpolarization-activated Ca2+ channel; ROS, reactive oxygen species.

4Present address: UMR INRA/UCBN 950 Ecophysiologie Végétale, Agronomie (EVA) & Nutrition NCS; ISBIO, Institut de Biologie Fondamentale et Appliquée, Université de CAEN Basse-Normandie, F-14032 Caen Cedex, France.

5Present address: Laboratory of Fermentation Engineering, College of Biology and Food Engineering, Dalian Institute of Light Industry, No. 1 Qinggongyuan, Dalian City 116034, PR China.

6Present address: Institut des Sciences du Végétal, UPR CNRS 2355, 1 avenue de la Terrasse, 91198 Gif-sur-Yvette cedex, France and Université Paris-Diderot, 2 place Jussieu, 75251 Paris cedex 05, France.

(Received June 2, 2008; Accepted August 10, 2008)
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