NADPH oxidase-mediated reactive oxygen species production: subcellular localization and reassessment of its role in plant defense.
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.
Lherminier
Jeannine
J
UMR PME INRA 1088/CNRS 5184/Université de Bourgogne, Dijon-cedex, France.
Elmayan
Taline
T
Fromentin
Jérôme
J
Elaraqui
Khadija Tantaoui
KT
Vesa
Simona
S
Morel
Johanne
J
Verrier
Jean-Louis
JL
Cailleteau
Bernard
B
Blein
Jean-Pierre
JP
Simon-Plas
Françoise
F
D002478:Cells, Cultured; D016373:DNA, Antisense; D005656:Fungal Proteins / Q000494:pharmacology; D006861:Hydrogen Peroxide / Q000032:analysis / Q000378:metabolism; D046529:Microscopy, Electron, Transmission; D010088:Oxidoreductases / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D018515:Plant Leaves / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D010940:Plant Proteins / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D030821:Plants, Genetically Modified / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000032:analysis / Q000378:metabolism*; D014026:Nicotiana / Q000187:drug effects / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure
D016373:DNA, Antisense; D005656:Fungal Proteins; D010940:Plant Proteins; D017382:Reactive Oxygen Species; D006861:Hydrogen Peroxide; C484277:NtrbohD protein, tobacco; D010088:Oxidoreductases