Light-dependent pH changes in leaves of C 3 plants: I. Recording pH changes in various cellular compartments by fluorescent probes

Chloroplasts, mesophyll protoplasts, cytoplasts, vacuoplasts, vacuoles and leaves were stained with pH-indicating fluorescent dyes of differing pK values. Fluorescence microscopy was used to obtain information on the intracellular and intercellular distribution of the probes. The kinetics of blue or green fluorescence emitted from chloroplasts, protoplasts, cytoplasts and leaves was measured during illumination with red light. The intensity of light used for fluorescence excitation was so low that it had little effect on photosynthesis. In leaves, fluorescence signals emitted from chloroplasts were small and usually insignificant compared to signals originating from the cytosol. Both indicated light-dependent alkalization and reversal of alkalization on darkening. Vacuolar signals were opposite in sign to cytosolic signals. They indicated acidification of the vacuole in the light-dark transient and reversal of this effect on darkening.