Plant Physiol. Drug Metab Dispos
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Plant Physiology 67:470-473 (1981)
© 1981 American Society of Plant Biologists

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Slow Passive Diffusion of Orthophosphate between Intact Isolated Chloroplasts and Suspending Medium 1

Gilles Mourioux and Roland Douce

Physiologie Cellulaire Végétale, Laboratoire de Biologie Végétale, Département de Recherche Fondamentale, Centre d'Etudes Nucléaires et Université Scientifique et Médicale de Grenoble, 85X 38041 Grenoble Cedex, France

Isolated spinach chloroplasts purified by isopycnic centrifugation in density gradients of Percoll were found to be highly intact, to be devoid of extrachloroplastic contaminations, and to retain a high rate of CO2-dependent O2 evolution.

When suspended in a medium which avoided rupture of the envelope, intact purified chloroplasts progressively lost their phosphate content by passive diffusion. This led to a slow decrease in the uptake of labeled 3-phosphoglyceric acid or orthophosphate (Pi) and in the rate of CO2-dependent O2 evolution by isolated chloroplasts. Under these conditions, there was a good correlation between the rate of CO2-dependent O2 evolution and the concentration of Pi in the stroma space. Addition of Pi to the suspending medium at a final concentration of 10 millimolar, which counterpoised the slow efflux of Pi from the chloroplasts, slowed considerably the decrease in the rate of CO2-dependent O2 evolution.

1 This work was supported in part by grants from the Centre National de la Recherche Scientifique (ERA 847: "Interactions plastescytoplasme-mitochondries") and the Délégation Générale à la Recherche Scientifique et Technique (contrat MB/P210).




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