Light-induced changes in chloroplasts of detached leaves of E. densa were investigated either by fixing leaves in 6% glutaraldehyde or by snap-freezing leaves and isolating chloroplasts after freeze-substitution in acetone. Individual chloroplasts were examined by electron probe analysis.Upon illumination (450 lux green light) chloroplasts in detached leaves flattened, with a reduction in volume of about 30% within the first second. During this initial phase of chloroplast contraction the calculated osmotic potential of chloroplasts fell from about -17 bars in the dark to -21 bars after 1 s of illumination, suggesting that the rapid contraction was not the result of an osmotic mechanism of water efflux brought about by an efflux of chloride, potassium, sodium, or calcium ions from chloroplasts. During the next hour of illumination chloroplast volume was reduced by only a further 5%, but the chloroplasts lost approximately 50% of their original ion content. The calculated osmotic potential of the chloroplasts rose to approximately -11 bars.During the first hour of illumination the average net efflux of ions (± standard error of the mean) across the chloroplast envelope was 32·6 ± 3·5 for chloride, 8·7 ± 0·8 for potassium, and 21·24 ± 1·4 p-equiv.cm-2 s-1 for sodium. The estimated efflux for calcium was approximately 8 p-equiv. cm-2 S-1. Typical concentrations of chloride, potassium, and sodium respectively in chloroplasts were 0·53, 0'24, and 0·31 equiv/l of chloroplast volume in dark-treated leaves and 0·32, 0·20, and 0·17 equiv/l in light-treated leaves. The chloride content of chloroplasts represents at least 20% of the total chloride content of dark-treated leaves. In the light the corresponding proportion is approximately 8%. The effects of CO2, ammonia, brucine, ouabain, and 3-p-chlorophenyl-1,I-dimethylurea on the in vivo ion and volume relations of E. densa chloroplasts are described.
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