Plant Physiology 92:293-301 (1990)
© 1990 American Society of Plant Biologists
Membranes and Bioenergetics
Inhibition of Zeaxanthin Formation and of Rapid Changes in Radiationless Energy Dissipation by Dithiothreitol in Spinach Leaves and Chloroplasts 1
Barbara Demmig-Adams2,
William W. Adams, III2,
Ulrich Heber,
Spidola Neimanis,
Klaus Winter,
Almuth Krüger,
Franz-Christian Czygan,
Wolfgang Bilger and
Olle Björkman
Institut für Botanik and Pharmazeutische Biologie Universität Würzburg, Mittlerer Dallenbergweg 64, 8700 Würzburg, Federal Republic of Germany,
Department of Plant Biology Carnegie Institution of Washington, Stanford, California 94305
Dithiothreitol, which completely inhibits the de-epoxidation of violaxanthin to zeaxanthin, was used to obtain evidence for a causal relationship between zeaxanthin and the dissipation of excess excitation energy in the photochemical apparatus in Spinicia oleracea L. In both leaves and chloroplasts, inhibition of zeaxanthin formation by dithiothreitol was accompanied by inhibition of a component of nonphotochemical fluorescence quenching. This component was characterized by a quenching of instantaneous fluorescence (Fo) and a linear relationship between the calculated rate constant for radiationless energy dissipation in the antenna chlorophyll and the zeaxanthin content. In leaves, this zeaxanthin-associated quenching, which relaxed within a few minutes upon darkening, was the major component of nonphotochemical fluorescence quenching determined in the light, i.e. it represented the `high-energy-state' quenching. In isolated chloroplasts, the zeaxanthin-associated quenching was a smaller component of total nonphotochemical quenching and there was a second, rapidly reversible high-energy-state component of fluorescence quenching which occurred in the absence of zeaxanthin and was not accompanied by Fo quenching. Leaves, but not chloroplasts, were capable of maintaining the electron acceptor, Q, of photosystem II in a low reduction state up to high degrees of excessive light and thus high degrees of nonphotochemical fluorescence quenching. When ascorbate, which serves as the reductant for violaxanthin de-epoxidation, was added to chloroplast suspensions, zeaxanthin formation at low photon flux densities was stimulated and the relationship between nonphotochemical fluorescence quenching and the reduction state in chloroplasts then became more similar to that found in leaves. We conclude that the inhibition of zeaxanthin-associated fluorescence quenching by dithiothreitol provides further evidence that there exists a close relationship between zeaxanthin and potentially photoprotective dissipation of excess excitation energy in the antenna chlorophyll.
2 Present address: Department of Environmental, Population, and Organismic Biology, University of Colorado, Boulder, Colorado 80309-0334.
1 Supported by the Deutsche Forschungsgemeinschaft (Forschergruppe Ökophysiologie and Sonderforschungsbereich 251) and by the Fonds der Chemischen Industrie. W. W. A. gratefully acknowledges the support of a fellowship from the Alexander von Humboldt Foundation.
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