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J. Biol. Chem., Vol. 282, Issue 29, 21160-21168, July 20, 2007

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Nitric Oxide Is Critical for Inducing Phosphatidic Acid Accumulation in Xylanase-elicited Tomato Cells^*

From the Instituto de Investigaciones Biológicas, Universidad Nacional de Mar del Plata, CC 1245, 7600 Mar del Plata, Argentina

Nitric Oxide (NO) is a second messenger related to development and (a)biotic stress responses in plants. We have studied the role of NO in signaling during plant defense responses upon xylanase elicitation. Treatment of tomato cell cultures with the fungal elicitor xylanase resulted in a rapid and dose-dependent NO accumulation. We have demonstrated that NO is required for the production of the lipid second messenger phosphatidic acid (PA) via the activation of the phospholipase C (PLC) and diacylglycerol kinase (DGK) pathway. Defense-related responses downstream of PA were studied. PA and, correspondingly, xylanase were shown to induce reactive oxygen species production. Scavenging of NO or inhibition of either the PLC or the DGK enzyme diminished xylanase-induced reactive oxygen species production. Xylanase-induced PLD1 and PR1 mRNA levels decreased when NO or PA production were compromised. Finally, we have shown that NO and PA are involved in the induction of cell death by xylanase. Treatment with NO scavenger cPTIO, PLC inhibitor U73122 [GenBank] , or DGK inhibitor R59022 [GenBank] diminished xylanase-induced cell death. On the basis of biochemical and pharmacological experimental results, we have shown that PLC/DGK-derived PA represents a novel downstream component of NO signaling cascade during plant defense.

Received for publication, February 8, 2007 , and in revised form, May 4, 2007.

^* This work was financially supported by Universidad Nacional de Mar del Plata (to A. M. L. and L. L.), Consejo Nacional de Investigaciones Científicas y Técnicas (to A. M. L. and L. L.), Fundación Antorchas (to A. M. L. and L. L.), Third World Academy of Sciences (to A. M. L.), Dutch Technology Foundation (STW; to A. t. H.) and Agencia Nacional de Promoción Científica y Tecnológica (to A. M. L. and L. L.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains supplemental Figs. S1 and S2.

¹ To whom correspondence should be addressed: Instituto de Investigaciones Biológicas, Universidad Nacional de Mar del Plata, CC 1245, 7600 Mar del Plata, Argentina. Tel.: 54-223-4753030; Fax: 54-223-4753150; E-mail: amlaxalt{at}mdp.edu.ar.

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