First published online November 18, 2005; 10.1104/pp.105.070763
Plant Physiology 139:1806-1820 (2005)
© 2005 American Society of Plant Biologists
ENVIRONMENTAL STRESS AND ADAPTATION TO STRESS
Characterization of Transformed Arabidopsis with Altered Alternative Oxidase Levels and Analysis of Effects on Reactive Oxygen Species in Tissue1,[W]
Ann L. Umbach*,
Fabio Fiorani2 and
James N. Siedow
Developmental, Cell, and Molecular Biology Group, Biology Department, Duke University, Durham, North Carolina 27708–1000
The alternative oxidase (AOX) of plant mitochondria transfers electrons from the ubiquinone pool to oxygen without energy conservation. AOX can use reductant in excess of cytochrome pathway capacity, preventing reactive oxygen species (ROS) formation from an over-reduced ubiquinone pool, and thus may be involved in acclimation to oxidative stresses. The AOX connection with mitochondrial ROS has been investigated only in isolated mitochondria and suspension culture cells. To study ROS and AOX in whole plants, transformed lines of Arabidopsis (Arabidopsis thaliana) were generated: AtAOX1a overexpressors, AtAOX1a anti-sense plants, and overexpressors of a mutated, constitutively active AtAOX1a. In the presence of KCN, leaf tissue of either mutant or wild-type AOX overexpressors showed no increase in oxidative damage, whereas anti-sense lines had levels of damage greater than those observed for untransformed leaves. Similarly, ROS production increased markedly in anti-sense and untransformed, but not overexpressor, roots with KCN treatment. Thus, AOX functions in leaves and roots, as in suspension cells, to ameliorate ROS production when the cytochrome pathway is chemically inhibited. However, in contrast with suspension culture cells, no changes in leaf transcript levels of selected electron transport components or oxidative stress-related enzymes were detected under nonlimiting growth conditions, regardless of transformation type. Further, a microarray study using an anti-sense line showed AOX influences outside mitochondria, particularly in chloroplasts and on several carbon metabolism pathways. These results illustrate the value of expanding AOX transformant studies to whole tissues.
1 This work was supported by the National Science Foundation (grant no. MCB–0091080 to J.N.S. and A.L.U.).
2 Present address: VIB-Ghent University, Plant Systems Biology, Technologiepark 927, B–9052 Ghent, Belgium.
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: Ann L. Umbach (umbacha{at}duke.edu).
[W] The online version of this article contains Web-only data.
Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.105.070763.
* Corresponding author; e-mail umbacha{at}duke.edu; fax 919–613–8177.
Received August 30, 2005;
returned for revision October 18, 2005;
accepted October 19, 2005.
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