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Originally published In Press as doi:10.1074/jbc.M304987200 on July 28, 2003

J. Biol. Chem., Vol. 278, Issue 40, 38921-38925, October 3, 2003
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The Water-Water Cycle Is Essential for Chloroplast Protection in the Absence of Stress*,

Ludmila Rizhsky {ddagger}, Hongjian Liang § and Ron Mittler § ¶

From the {ddagger}Department of Biology, Technion-Israel Institute of Technology, Technion City, Haifa 32000, Israel and the §Department of Botany, Plant Sciences Institute, Iowa State University, Ames, Iowa 50011

Maintaining electron flow through the photosynthetic apparatus, even in the absence of a sufficient amount of NADP+ as an electron acceptor, is essential for chloroplast protection from photooxidative stress. At least two different pathways are thought to participate in this process, i.e. cyclic electron flow and the water-water cycle. Although the function of the water-water cycle was inferred from a number of biochemical and physiological studies, genetic evidence for the function of this cycle is very limited. Here we show that knockdown Arabidopsis plants with suppressed expression of the key water-water cycle enzyme, thylakoid-attached copper/zinc superoxide dismutase (KD-SOD), are suppressed in their growth and development. Chloroplast size, chlorophyll content, and photosynthetic activity were also reduced in KD-SOD plants. Microarray analysis of KD-SOD plants, grown under controlled conditions, revealed changes in transcript expression consistent with an acclimation response to light stress. Although a number of transcripts involved in the defense of plants from oxidative stress were induced in KD-SOD plants, and seedlings of KD-SOD plants were more tolerant to oxidative stress, these mechanisms were unable to compensate for the suppression of the water-water cycle in mature leaves. Thus, the localization of copper/zinc superoxide dismutase at the vicinity of photosystem I may be essential for its function. Our studies provide genetic evidence for the importance of the water-water cycle in protecting the photosynthetic apparatus of higher plants from photooxidative damage.


Received for publication, May 13, 2003 , and in revised form, July 21, 2003.

* This work was supported by funding from the Plant Sciences Institute at Iowa State University, the Biotechnology Council of Iowa State University, the College of Liberal Arts and Sciences at Iowa State University, the Israeli Academy of Science, and the Fund for the Promotion of Research at the Technion. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains supplementary Figs. 1 and 2 and supplementary Tables I, II, and III.

To whom correspondence should be addressed: Dept. of Biochemistry, University of Nevada, Reno, NV 89557. Tel.: 775-784-6031; Fax: 775-784-1419; E-mail: ronm{at}unr.edu.


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