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Plant and Cell Physiology Advance Access originally published online on September 30, 2006
Plant and Cell Physiology 2006 47(11):1509-1519; doi:10.1093/pcp/pcl016
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© The Author 2006. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Changes in Plant Mitochondrial Electron Transport Alter Cellular Levels of Reactive Oxygen Species and Susceptibility to Cell Death Signaling Molecules

Sasan Amirsadeghi1, Christine A. Robson1, Allison E. McDonald and Greg C. Vanlerberghe*

Department of Life Sciences and Department of Cell and Systems Biology, University of Toronto Scarborough, 1265 Military Trail, Toronto, ON, Canada M1C1A4

* Corresponding author: E-mail, gregv{at}utsc.utoronto.ca; Fax, +1-416-287-7642.


  Abstract

Transgenic tobacco (Nicotiana tabacum) lacking mitochondrial alternative oxidase (AOX) have been compared with wild-type (Wt) tobacco using two different systems, either suspension cell cultures or leaves. In both systems, a lack of AOX was accompanied by an increase in some anti-oxidant defenses, consistent with the hypothesis that a lack of AOX increases the mitochondrial generation of reactive oxygen species (ROS). In most cases, this increase in anti-oxidant defenses could more than offset the presumed increased rate of ROS generation, resulting paradoxically in a lower steady-state level of ROS than was found in Wt leaves or suspension cells. We also found that the amount of cell death induced by salicylic acid or nitric oxide correlated strongly with the level of ROS (irrespective of the level of AOX), while death induced by azide was dependent upon the presence or absence of AOX. These results suggest that susceptibility to cell death by signaling molecules (salicylic acid and nitric oxide) is dependent upon the steady-state cellular level of ROS and that AOX levels clearly contribute to this steady state, perhaps by influencing the rate of mitochondrial-generated ROS and hence the cellular level of anti-oxidant defenses.

Keywords: Alternative oxidase - Anti-oxidant defenses - Cell death - Mitochondrial electron transport - Reactive oxygen species - Transgenic tobacco

Abbreviations: AA, antimycin A; AOX, alternative oxidase; APx, ascorbate peroxidase; CAT, catalase; CuZnSOD, copper zinc superoxide dismutase; cyt, cytochrome; DAB, 3,3'-diaminobenzidine; DCFH-DA, 2'7'-dichlorodihydrofluorescin diacetate; ETC, electron transport chain; FeSOD, iron superoxide dismutase; GPx, glutathione peroxidase; H2O2, hydrogen peroxide; mROS, mitochondrial reactive oxygen species; MnSOD, manganese superoxide dismutase; NBT, nitrotetrazolium blue chloride; N3, azide; NO, nitric oxide; PCD, programmed cell death; PTOX, plastid terminal oxidase; ROS, reactive oxygen species; RT–PCR, reverse transcription–PCR; SA, salicylic acid; SNP, sodium nitroprusside; Wt, wild-type.

1 These authors contributed equally to this work.

(Received July 11, 2006; Accepted September 23, 2006)
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