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PLANT PHYSIOLOGY , Vol 108, Issue 1 277-283, Copyright © 1995 by American Society of Plant Biologists
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WHOLE PLANT, ENVIRONMENTAL, AND STRESS PHYSIOLOGY |
Measurements of in Vivo Ubiquinone Reduction Levels in Plant Cells
A. M. Wagner and M. J. Wagner
Department of Physiology and Biochemistry of Plants, Vrije Universiteit, De Boelelaan 1087, 1081 HV Amsterdam, The Netherlands
A method is described for the determination of in vivo ubiquinone (UQ)
reduction levels in nongreen tissues by extraction and subsequent detection
of ubiquinone-10 and ubiquinol-10 with high-performance liquid
chromatography. In Petunia hybrida cell suspensions UQ reduction remained
at a stable level of about 60%, despite the changing conditions during the
batch culture (from excess sugar to starvation) and the concomitant
variations in respiration. Also, in the presence of uncoupler, which causes
a large increase in respiration via both the cytochrome pathway and the
alternative pathway, UQ reduction levels stayed at 60%. In mitochondria
isolated from these cells, activity of the alternative pathway was only
observed at UQ reduction levels higher than 80%. It is proposed that in
vivo the relationship between UQ reduction and the activity of the
alternative oxidase is modulated by mechanisms such as thiol modifications
and accumulation of organic acids. Accordingly, pyruvate concentration in
P. hybrida cells increased in the presence of uncoupler.
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