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CELL BIOLOGY AND SIGNAL TRANSDUCTION

Phospholipase D Activation Is an Early Component of the Salicylic Acid Signaling Pathway in Arabidopsis Cell Suspensions^1,[W],[OA]

Ondej Krinke, Matyá Flemr, Chantal Vergnolle, Sylvie Collin, Jean-Pierre Renou, Ludivine Taconnat, Agnès Yu, Lenka Burketová, Olga Valentová, Alain Zachowski and Eric Ruelland^*

UPMC Univ Paris 06, Unité de Recherche 5, Centre National de la Recherche Scientifique, Equipe d'Accueil Conventionnée 7180, Laboratoire de Physiologie Cellulaire et Moléculaire des Plantes, F–94200 Ivry-sur-Seine, France (O.K., M.F., C.V., S.C., A.Z., E.R.); Institute of Chemical Technology, Department of Biochemistry and Microbiology, Prague, 166 28 Czech Republic (O.K., M.F., O.V.); Unité Mixte de Recherche Institut National de la Recherche Agronomique 1165, Centre National de la Recherche Scientifique 8114, Unité de Recherche en Génomique Végétale, F–91057 Evry, France (J.-P.R., L.T., A.Y.); and Academy of Sciences of the Czech Republic, Institute of Experimental Botany, Prague, 160 00 Czech Republic (L.B.)

Salicylic acid (SA) plays a central role in defense against pathogen attack, as well as in germination, flowering, senescence, and the acquisition of thermotolerance. In this report we investigate the involvement of phospholipase D (PLD) in the SA signaling pathway. In presence of exogenous primary alcohols, the production of phosphatidic acid by PLD is diverted toward the formation of phosphatidylalcohols through a reaction called transphosphatidylation. By in vivo metabolic phospholipid labeling with ³³P_i, PLD activity was found to be induced 45 min after addition of SA. We show that incubation of Arabidopsis (Arabidopsis thaliana) cell suspensions with primary alcohols inhibited the induction of two SA-responsive genes, PATHOGENESIS-RELATED1 and WRKY38, in a dose-dependent manner. This inhibitory effect was more pronounced when the primary alcohols were more hydrophobic. Secondary or tertiary alcohols had no inhibitory effect. These results provide compelling arguments for PLD activity being upstream of the induction of these genes by SA. A subsequent study of n-butanol effects on the SA-responsive transcriptome identified 1,327 genes differentially expressed upon SA treatment. Strikingly, the SA response of 380 of these genes was inhibited by n-butanol but not by tert-butanol. A detailed analysis of the regulation of these genes showed that PLD could act both positively and negatively, either on gene induction or gene repression. The overlap with the previously described phosphatidylinositol-4-kinase pathway is discussed.

The author responsible for the distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Eric Ruelland (eric.ruelland{at}upmc.fr).

^[W] The online version of this article contains Web-only data.

^[OA] Open access articles can be viewed online without a subscription.

www.plantphysiol.org/cgi/doi/10.1104/pp.108.133595

^* Corresponding author; e-mail eric.ruelland{at}upmc.fr.

Received December 11, 2008; accepted March 17, 2009; published March 20, 2009.

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Plant Physiol. 2009 150: 1-2. [Full Text]