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First published online April 22, 2009; 10.1104/pp.109.138230 Plant Physiology 150:801-814 (2009) © 2009 American Society of Plant Biologists Reactive Oxygen Species Are Involved in Brassinosteroid-Induced Stress Tolerance in Cucumber1,[W]Department of Horticulture, Zhejiang University, Hangzhou 310029, China (X.-J.X., Y.-J.W., Y.-H.Z., Y.T., W.-H.M., K.S., J.-Q.Y.); Key Laboratory of Horticultural Plant Growth, Development, and Biotechnology, Agricultural Ministry of China, Hangzhou 310029, China (Y.-H.Z., J.-Q.Y.); Department of Applied Biological Chemistry, University of Tokyo, Bunkyo Ku, Tokyo 1138657, Japan (T.A.); and Department of Botany and Plant Pathology, Purdue University, West Lafayette, Indiana 47907–2054 (Z.C.)
Brassinosteroids (BRs) induce plant tolerance to a wide spectrum of stresses. To study how BR induces stress tolerance, we manipulated the BR levels in cucumber (Cucumis sativus) through a chemical genetics approach and found that BR levels were positively correlated with the tolerance to photo-oxidative and cold stresses and resistance to Cucumber mosaic virus. We also showed that BR treatment enhanced NADPH oxidase activity and elevated H2O2 levels in apoplast. H2O2 levels were elevated as early as 3 h and returned to basal levels 3 d after BR treatment. BR-induced H2O2 accumulation was accompanied by increased tolerance to oxidative stress. Inhibition of NADPH oxidase and chemical scavenging of H2O2 reduced BR-induced oxidative and cold tolerance and defense gene expression. BR treatment induced expression of both regulatory genes, such as RBOH, MAPK1, and MAPK3, and genes involved in defense and antioxidant responses. These results strongly suggest that elevated H2O2 levels resulting from enhanced NADPH oxidase activity are involved in the BR-induced stress tolerance.
1 This work was supported by the National Basic Research Program of China (grant no. 2009CB119000), the National Natural Science Foundation of China (grant nos. 3050344 and 30671428), and the Program for Promotion of Basic Research Activities for Innovative Bioscience (PROBRAIN). 2 These authors contributed equally to the article. The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Jing-Quan Yu (jqyu{at}zju.edu.cn). [W] The online version of this article contains Web-only data. www.plantphysiol.org/cgi/doi/10.1104/pp.109.138230 * Corresponding author; e-mail jqyu{at}zju.edu.cn. Received March 8, 2009; accepted April 20, 2009; published April 22, 2009.
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