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Published on February 24, 2006; 10.1104/pp.105.074690

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Received November 24, 2005
Returned for revision December 16, 2005
Accepted February 10, 2006

Adducts of oxylipin electrophiles to glutathione reflect a 13 specificity of the downstream lipoxygenase pathway in the tobacco hypersensitive response

Céline Davoine , Olivier Falletti , Thierry Douki , Gilles Iacazio , Najla Ennar , Jean-Luc Montillet , and Christian Triantaphylidès *

Laboratoire de Radiobiologie Végétale, DSV-Département d'Ecophysiologie Végétale et de Microbiologie, CEA-Cadarache, 13108 Saint-Paul Lez Durance Cedex, France
Laboratoire "Lésions des Acides Nucléiques", Service de Chimie Inorganique et Biologique, DSM-Département de Recherche Fondamentale sur la Matière Condensée, UMR-E 3 CEA-UJF, CEA-Grenoble, 38054 Grenoble Cedex 9, France
Laboratoire de Bioinorganique Structurale, Case 432, UMR CNRS 6517 Chimie, Biologie et Radicaux libres, Faculté des Sciences de St Jérôme, 13397 Marseille Cedex 20, France

* Corresponding author; email: ctriantaphylid{at}cea.fr.

The response to Reactive Electrophile Species (RES) is now considered as part of the plant response to pathogen and insect attacks. Thanks to a previous established HPLC tandem mass spectrometry methodology, we have investigated the production of oxylipin RES adducts to GSH during the Hypersensitive Response (HR) of plants. We have observed that RES conjugation to glutathione (GSH) in tobacco leaves is facile and non-specific. In cryptogein-elicited tobacco leaves, we show that the oxylipin RES adducts to GSH are produced in correlation with GSH consumption, increase in glutathione-S-transferase activity and the appearance of the cell death symptoms. In this model, the adducts arise mainly from the downstream 13 lipoxygenase (LOX) metabolism, although the induced 9 LOX pathway leads massively to the accumulation of upstream metabolites. The main adducts were obtained from 2-hexenal and 12-oxo-phytodienoic acid (12-OPDA). They accumulate transiently as 1-hexanol-3-GSH, a reduced adduct, and 12-OPDA-GSH, respectively. RES conjugation does not initiate cell death but explains part of the GSH depletion which accompanies HR cell death. The nature of these GSH conjugates shows the key role played by the 13 LOX pathway in RES signaling in the tobacco HR.




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