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NADPH Oxidase-Mediated Reactive Oxygen Species Production: Subcellular Localization and Reassessment of Its Role in Plant Defense

July 2009 , Volume 22 , Number  7
Pages  868 - 881

Jeannine Lherminier,1 Taline Elmayan,1 Jérôme Fromentin,1 Khadija Tantaoui Elaraqui,1 Simona Vesa,1 Johanne Morel,1 Jean-Louis Verrier,2 Bernard Cailleteau,2 Jean-Pierre Blein,1 and Françoise Simon-Plas1

1UMR PME INRA 1088/CNRS 5184/Université de Bourgogne, 17 Rue Sully, BP 86510 21065, Dijon-cedex, France; 2ALTADIS-Institut du Tabac 769 route de Sainte-Alvère, 24100 Bergerac, France

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Accepted 24 February 2009.

Chemiluminescence detection of reactive oxygen species (ROS) triggered in tobacco BY-2 cells by the fungal elicitor cryptogein was previously demonstrated to be abolished in cells transformed with an antisense construct of the plasma membrane NADPH oxidase, NtrbohD. Here, using electron microscopy, it has been confirmed that the first hydrogen peroxide production occurring a few minutes after challenge of tobacco cells with cryptogein is plasma membrane located and NtrbohD mediated. Furthermore, the presence of NtrbohD in detergent-resistant membrane fractions could be associated with the presence of NtrbohD-mediated hydrogen peroxide patches along the plasma membrane. Comparison of the subcellular localization of ROS in wild-type tobacco and in plants transformed with antisense constructs of NtrbohD revealed that this enzyme is also responsible for the hydrogen peroxide production occurring at the plasma membrane after infiltration of tobacco leaves with cryptogein. Finally, the reactivity of wild-type and transformed plants to the elicitor and their resistance against the pathogenic oomycete Phytophthora parasitica were examined. NtrbohD-mediated hydrogen peroxide production does not seem determinant for either hypersensitive response development or the establishment of acquired resistance but it is most likely involved in the signaling pathways associated with the protection of the plant cell.

© 2009 The American Phytopathological Society