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Metabolism and Enzymology

Photosynthesis by Intact Isolated Chloroplasts on Solid Support

Institute of Botany and Botanical Garden, Faculty of Science, University of Belgrade, 11000 Belgrade, Yugoslavia, Research Institute for Photosynthesis, University of Sheffield, Sheffield S10 2TN, United Kingdom

A new approach to measurements of photosynthesis by isolated chloroplasts has been devised. Intact isolated chloroplasts were trapped in the cavities of membrane filters. The thin layers of chloroplasts so obtained were assayed for O₂ evolution and CO₂ assimilation in leaf-chambers. Photosynthetic gas exchange could be demonstrated to take place either in a closed or a flow-through system. The chloroplasts were morphologically intact as shown by light or scanning electron microscopy and displayed stable rates of photosynthesis in the presence of phosphate and alkaline phosphatase. The methods described open the way to in vitro measurement of photosynthesis, by chloroplasts under conditions more closely resembling those in leaves.