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First published online December 17, 2004; 10.1105/tpc.104.026971

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The Plant Cell 17:268-281 (2005)
© 2005 American Society of Plant Biologists

Cytosolic Ascorbate Peroxidase 1 Is a Central Component of the Reactive Oxygen Gene Network of Arabidopsis{boxw}

Sholpan Davletovaa, Ludmila Rizhskyb, Hongjian Liangb, Zhong Shengqiangb, David J. Oliverb, Jesse Coutua, Vladimir Shulaevc, Karen Schlauchd and Ron Mittlera,1

a Department of Biochemistry and Molecular Biology, University of Nevada, Reno, Nevada 89557
b Department of Genetics and Developmental and Cell Biology, Iowa State University, Ames, Iowa 50011
c Virginia Bioinformatics Institute, Blacksburg, Virginia 24061
d Center for Biomedical Genomics and Informatics, George Mason University, Manassas, Virginia 20110

1 To whom correspondence should be addressed. E-mail ronm{at}unr.edu; fax 775-784-6911.

Reactive oxygen species (ROS), such as O2– and H2O2, play a key role in plant metabolism, cellular signaling, and defense. In leaf cells, the chloroplast is considered to be a focal point of ROS metabolism. It is a major producer of O2– and H2O2 during photosynthesis, and it contains a large array of ROS-scavenging mechanisms that have been extensively studied. By contrast, the function of the cytosolic ROS-scavenging mechanisms of leaf cells is largely unknown. In this study, we demonstrate that in the absence of the cytosolic H2O2-scavenging enzyme ascorbate peroxidase 1 (APX1), the entire chloroplastic H2O2-scavenging system of Arabidopsis thaliana collapses, H2O2 levels increase, and protein oxidation occurs. We further identify specific proteins oxidized in APX1-deficient plants and characterize the signaling events that ensue in knockout-Apx1 plants in response to a moderate level of light stress. Using a dominant-negative approach, we demonstrate that heat shock transcription factors play a central role in the early sensing of H2O2 stress in plants. Using knockout plants for the NADPH oxidase D protein (knockout-RbohD), we demonstrate that RbohD might be required for ROS signal amplification during light stress. Our study points to a key role for the cytosol in protecting the chloroplast during light stress and provides evidence for cross-compartment protection of thylakoid and stromal/mitochondrial APXs by cytosolic APX1.




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