The Fabrication of H+-Selective Liquid-Membrane Micro-electrodes for Use in Plant Cells

The hydrogen ion-sensitive liquid-membrane micro-electrode,as described by Amman, Lanter, Steiner, Schulthess, Shijo, andSimon (1981) has been developed further and made applicablefor turgescent plant cells even with tough cell walls, by treatmentwith polyvinylchloride (PVC). Such an electrode is slower (t=5–10s) than the untreated electrode (t=2–6 s), but displays55–59 mV/pH-unit between pH 4·3 and 9·0,and is almost insensitive towards different buffers and K⁺.The electrodes are usable on more than one cell and have stillgood recalibration properties. Testing the electrodes on 11different cell types of Riccia fluitans, Sinapis alba, Zea mays,Avena sativa, Kalanchöe daigremontiana, Lemna gibba andChara corallina, we find the internal pH slightly alkaline (7·1–7·6)in ten cases (exception: old rhizoids of Riccia, pH₁=4·8).From that we conclude that the pH-electrode measures in thecytosol. According to the different plant material, severalprocedures for internal pH-measurements are presented and supportedby data: Application of cyanide and acetic acid causes internalacidification. Light-off transiently alkalinazes, light-on transientlyacidifies the internal pH. The advantages and limitations ofthe method are critically discussed, and it is concluded thatthis electrode is a powerful tool in plant physiology. Key words: Internal pH, pH-sensitive micro-electrode