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Evidence for a significant contribution by peroxidase-mediated O2 uptake to root respiration of Brachypodium pinnatum

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Abstract

This study describes the O2 uptake characteristics of intact roots of Brachypodium pinnatum. In the presence of 25 mM salicylhydroxamic acid (SHAM), concentrations of KCN below 3.5 νM had no effect on the rate of root respiration, whereas in the absence of 25 mM SHAM a significant inhibition of approx. 18% was observed. This indicates that an O2-consuming reaction, not associated with the cytochrome pathway, the alternative pathway or the “residual component”, operates in the absence of any inhibitors in roots of B. pinnatum. We demonstrate here that this fourth O2-consuming reaction is mediated by a peroxidase. A peroxidase which catalyzed O2 reduction in the presence of NADH was readily washed from the roots of B. pinnatum. This peroxidase was stimulated by 5 mM SHAM, whereas ascorbic acid, catalase, catechol, gentisic acid, low concentrations potassium cyanide (3.5 μM), sodium azide, sodium sulfide, superoxide dismutase and high concentrations SHAM (25 mM) inhibited this reaction. Except for high concentrations of SHAM and concentrations of KCN higher than approx. 3.5 μM, these effectors could not be used to inhibit the peroxidase-mediated O2 uptake in intact roots of B. pinnatum. Concentrations of SHAM below 10 mM stimulated O2 uptake up to 15% of the control rate, depending on concentration, whereas 25 mM SHAM inhibited O2 uptake by 35%. The stimulation at low concentrations resulted from a SHAM-stimulated peroxidase activity, whereas 25 mM SHAM completely inhibited both the peroxidase-mediated O2 uptake and the activity of the alternative pathway. A method is presented for determining the relative contributions of each of the four O2-consuming reactions, i.e. the cytochrome pathway, the alternative pathway, the “residual component” and the peroxidase-mediated O2 uptake. The peroxidase-mediated O2 uptake contributed 21% to the total rate of oxygen uptake in roots of B. pinnatum, the cytochrome pathway contributed 41%, the alternative pathway 14% and the “residual component” 24%.

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Abbreviations

SHAM:

salicylhydroxamic acid

SOD:

superoxide dismutase

Valt :

capacity of the alternative pathway

vcyt :

cytochrome-mediated respiration

vpod :

peroxidase-mediated consumption of O2 in the absence of SHAM and KCN

vres :

residual respiration in the presence of SHAM and KCN

vt :

total rate of O2 consumption in the absence of inhibitors

ϱ:

fraction of the capacity of the alternative pathway that is actually engaged

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We wish to thank our colleagues Edwin Kraus and Hendrik Poorter for their critical reading of the manuscript. The contribution of A. van der Werf was financially supported by the Foundation for Fundamental Biological Research (BION), which is subsidized by the Netherlands Organization for Scientific Research (NWO).

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van der Werf, A., Raaimakers, D., Poot, P. et al. Evidence for a significant contribution by peroxidase-mediated O2 uptake to root respiration of Brachypodium pinnatum . Planta 183, 347–352 (1991). https://doi.org/10.1007/BF00197732

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  • DOI: https://doi.org/10.1007/BF00197732

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