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AOBPreview published online on July 18, 2006

Annals of Botany, doi:10.1093/aob/mcl162
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© The Author 2006. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org
Received April 6, 2006
Revised May 10, 2006
Accepted June 1, 2006

Article

Evaluation of Metabolic Alteration in Transgenic Rice Overexpressing Dihydroflavonol-4-reductase

HIDEYUKI TAKAHASHI 1, MITSUNORI HAYASHI 1, FUMIYUKI GOTO 2, SHIGERU SATO 3, TOMOYOSHI SOGA 3, TAKAAKI NISHIOKA 3, MASARU TOMITA 3, MAKI KAWAI-YAMADA 1, and HIROFUMI UCHIMIYA 4 *

1 Institute of Molecular and Cellular Biosciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-0032, Japan
2 Plant Molecular Biology Group, Laboratory of Environmental Science, Central Research Institute Electric Power Industry, 1646 Abiko, Chiba 270-1194, Japan
3 Institute for Advanced Biosciences, University of Keio, Tsuruoka, Yamagata 997-0017, Japan
4 Institute of Molecular and Cellular Biosciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-0032, Japan; Iwate Biotechnology Research Center, Kitakami, Iwate 024-0003, Japan

* To whom correspondence should be addressed.
HIROFUMI UCHIMIYA, E-mail: uchimiya{at}iam.u-tokyo.ac.jp


  Abstract

Background and Aims Previous studies have shown that transgenic rice plants overexpressing YK1, which possesses dihydroflavonol-4-reductase (DFR) activity, showed biotic and abiotic stress tolerance. High throughput profiles of metabolites have also been shown in such transgenic plants by Fourier transform ion cyclotron mass spectrometry. In this study, capillary electrophoresis mass spectrometry analysis (CE/MS) was employed to identify precise metabolites such as organic acids, amino acids and sugars.

Methods Using CE/MS, we analysed several metabolites of glycolysis, the tricarboxylic acid (TCA) cycle and the pentose phosphate pathway. In addition, the concentrations of sugars and ion were quantified.

Key Results In YK1 (DFR)-overexpressing plants, the concentrations of cis-aconitate, isocitrate and 2-oxoglutarate were higher in leaves, whereas those of fructose-1, 6-bisphosphate and glyceraldehyde-3-phosphate were lower in roots. In seeds, the amounts of free amino acids and metals were altered, whereas sugars in seeds were kept constant. In YK1 calli, an approx. 3-fold increase in glutathione was observed, whereas the activities of glutathione peroxidase and glutathione reductase were concomitantly increased.

Conclusions The overexpression of YK1 (DFR) was associated with slight changes in the amounts of several metabolites analysed in whole plants, whilst glutathione derivatives were substantially increased in suspension-cultured cells.

Keywords: Metabolome, dihydroflavonol-4-reductase, capillary electrophoresis, mass spectrometry, rice, Oryza sativa.
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