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Manipulating cytoplasmic pH under anoxia: A critical test of the role of pH in the switch from aerobic to anaerobic metabolism

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Abstract

Ethanol production by maize (Zea mays L.) root tips, measured by an enzymic assay of the suspending medium, was correlated with changes in the cytoplasmic pH, determined by in-vivo 31P nuclear magnetic resonance (NMR) spectroscopy, following the onset of anoxia. Strong evidence for the role of the cytoplasmic pH in triggering the switch to ethanol production under anoxia was obtained by: (i) varying the pH of the suspending medium between pH 4 and pH 10; and (ii) using the permeant weak base methylamine to combat the acidification of the cytoplasm induced by the anoxic conditions. Experimentally, it proved to be much easier to manipulate the cytoplasmic pH under anoxia after the pH had stabilised, rather than during the initial rapid acidification that occurred following the onset of anoxia, and in the presence of methylamine, it was possible to impose a normal aerobic cytoplasmic pH value on tissue that was metabolising anaerobically. By this means it was possible to demonstrate the reversibility of the pH effect on ethanol production under anoxia and thus to provide good evidence in support of the biochemical pH-stat model of the anoxic response. The NMR measurement of the cytoplasmic pH in the presence of methylamine was achieved by using a manganese pretreatment technique to eliminate interference between the cytoplasmic and vacuolar Pi signals, and it seems likely that the experimental approach used here will have further applications in studies of the metabolic response to anoxia.

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Abbreviations

Caps:

3-(cyclohexylamino)-1-propane sulphonic acid

Mes:

2-(N-morpholino)-ethane sulphonic acid

NMR:

nuclear magnetic resonance

Pi:

inorganic phosphate

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Authors and Affiliations

  1. Department of Plant Sciences, University of Oxford, South Parks Road, OX1 3RB, Oxford, UK

    G. G. Fox, N. R. McCallan & R. G. Ratcliffe

Authors
  1. G. G. Fox
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  2. N. R. McCallan
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  3. R. G. Ratcliffe
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Corresponding author

Correspondence to R. G. Ratcliffe.

Additional information

We acknowledge the financial support of the Agricultural and Food Research Council and G.G.F. acknowledges the receipt of a Research Fellowship from the Royal Commission for the Exhibition of 1851.

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Fox, G.G., McCallan, N.R. & Ratcliffe, R.G. Manipulating cytoplasmic pH under anoxia: A critical test of the role of pH in the switch from aerobic to anaerobic metabolism. Planta 195, 324–330 (1995). https://doi.org/10.1007/BF00202588

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  • DOI: https://doi.org/10.1007/BF00202588

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