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Plant and Cell Physiology Advance Access originally published online on December 2, 2008
Plant and Cell Physiology 2009 50(1):26-36; doi:10.1093/pcp/pcn184
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© The Author 2008. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved.
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and the Japanese Society of Plant Physiologists are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions@oxfordjournals.org

Special Issue - Regular Paper

Boron Nutrition of Tobacco BY-2 Cells. V. Oxidative Damage is the Major Cause of Cell Death Induced by Boron Deprivation

Taichi Koshiba, Masaru Kobayashi* and Toru Matoh

Laboratory of Plant Nutrition, Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto, 606-8502 Japan

*Corresponding author: E-mail, masaru{at}kais.kyoto-u.ac.jp; Fax, +81-75-753-6128.


  Abstract

Boron (B) is an essential micronutrient for vascular plants. However, it remains unclear how B deficiency leads to various metabolic disorders and cell death. To understand this mechanism, we analyzed the physiological changes in suspension-cultured tobacco (Nicotiana tabacum) BY-2 cells upon B deprivation. When 3-day-old cells were transferred to B-free medium, cell death was detectable as early as 12 h after treatment. The B-deprived cells accumulated more reactive oxygen species and lipid peroxides than control cells, and showed a slight but significant decrease in the cellular ascorbate pool. Supplementing the media with lipophilic antioxidants effectively suppressed the death of B-deprived cells, suggesting that the oxidative damage is the immediate and major cause of cell death under B deficiency. Dead cells in B-free culture exhibited a characteristic morphology with a shrunken cytoplasm, which is often seen in cells undergoing programmed cell death (PCD). However, they did not display other hallmarks of PCD such as internucleosomal DNA fragmentation, decreased ascorbate peroxidase expression and protection from death by cycloheximide. These results suggest that the death of tobacco cells induced by B deprivation is not likely to be a typical PCD.

Keywords: Boron deficiency - Cell death - Necrosis - Oxidative damage - Tobacco

Abbreviations: ASC, ascorbate; B, boron; BHA, butylated hydroxyanisole; CHX, cycloheximide; DHA, dehydroas-corbate; DHE, dihydroethidium; FDA, fluorescein diacetate; GSH, reduced glutathione; PCD, programmed cell death; PI, propidium iodide; RG-II, rhamnogalacturonan II; ROS, reactive oxygen species; RT–PCR, reverse transcription–PCR; TBARS, thiobarbituric acid reactive substances; TFA, trifluoroacetic acid.

(Received October 31, 2008; Accepted November 26, 2008)
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