Protoplasts as a tool for isolating functional chloroplasts from leaves

Rathnam, C K M and Edwards, G E (1976) Protoplasts as a tool for isolating functional chloroplasts from leaves. Plant and Cell Physiology, 17 (1). pp. 177-186.

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Leaf protoplasts from various grasses can be used for isolating chloroplasts with high photosyndietic activity. The protoplasts were stable for more than 20 hr during which time chloroplasts could be isolated from protoplasts without any loss of original CO2 fixation capacity (100–157 μmoles/mg chl-hr). Using Triticum aestiuum to optimize assay conditions, the pH optimum for CO2 fixation by the chloroplasts isolated from protoplasts was between 8.2 and 8.6. Magnesium (0.75 mM) was required for maximum CO2 fixation by the isolated chloroplasts and sodium ascorbate in the medium allowed a more linear increase in CO2 fixation with time. Based upon 14CO2 fixation and ferricyanide-dependent oxygen evolution as criteria of intactness, chloroplasts from protoplasts exhibited a high degree of intactness compared to those obtained by mechanical grinding. Chloroplasts isolated from grass leaves by mechanical grinding had a relatively low capacity for endogenous CO2 fixation and required addition of ribose-5-phosphate and ADP for maximum activity.

Item Type: Article
Author Affiliation: University of Wisconsin Madison, Wisconsin 53706, U. S. A.
Subjects: Plant Production
Plant Physiology and Biochemistry > Plant Physiology
Divisions: General
Depositing User: Library ICRISAT-InfoSAT
Date Deposited: 23 Feb 2012 13:59
Last Modified: 23 Feb 2012 13:59
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