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Relationships among violaxanthin deepoxidation, thylakoid membrane conformation, and nonphotochemical chlorophyll fluorescence quenching in leaves of cotton (Gossypium hirsutum L.)

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Abstract

The kinetics and temperature dependencies of development and relaxation of light-induced absorbance changes caused by deepoxidation of violaxanthin to antheraxanthin and zeaxanthin (ΔZ; peak at 506 nm) and by light scattering (ΔS; peak around 540 nm) as well as of nonphotochemical quenching of chlorophyll fluorescence (NPQ) were followed in cotton leaves. Measurements were made in the absence and the presence of dithiothreitol (DTT), an inhibitor of violaxanthin deepoxidase. The amount of NPQ was calculated from the Stern-Volmer equation. A procedure was developed to correct gross AS (ΔSg) for absorbance changes around 540 nm that are due to a spectral overlap with ΔZ. This protocol isolated the component which is caused by light-scattering changes alone (ΔSn). In control leaves, the kinetics and temperature dependence of the initial rate of rise in ΔSn that takes place upon illumination, closely matched that of ΔZ. Application of DTT to leaves, containing little zeaxanthin or antheraxanthin, strongly inhibited both ΔSn and NPQ, but DTT had no inhibitory effect in leaves in which these xanthophylls had already been preformed, showing that the effect of DTT on ΔAn and NPQ results solely from the inhibition of violaxanthin deepoxidation. The rates and maximum extents of ΔSn and NPQ therefore depend on the amount of zeaxanthin (and/or antheraxanthin) present in the leaf. In contrast to the situation during induction, relaxation of ΔZ upon darkening was much slower than the relaxation of ΔSn and NPQ. The relaxation of ΔSn and NPQ showed quantitatively similar kinetics and temperature dependencies (Q10=2.4). These results are consistent with the following hypotheses: The increase in lumen-proton concentration resulting from thylakoid membrane energization causes deepoxidation of violaxanthin to antheraxanthin and zeaxanthin. The presence of these xanthophylls is not sufficient to cause ΔSn or NPQ but, together with an increased lumen-proton concentration, these xanthophylls cause a conformational change, reflected by ΔSn. The conformational change facilititates nonradiative energy dissipation, thereby causing NPQ. Membrane energization is prerequisite to conformational changes in the thylakoid membrane and resultant nonradiative energy dissipation but the capacity for such changes in intact leaves is quite limited unless zeaxanthin (and/or antheraxanthin) is present in the membrane. The sustained ΔSn and NPQ levels that remain after darkening may be attributable to a sustained high lumen-proton concentration.

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Abbreviations

A:

antheraxanthin

DTT:

dithiothreitol

F, Fm :

chlorophyll fluorescence yield at actual, full closure of the PSII centers

NPQ:

nonphotochemical chlorophyll fluorescence quenching

PFD:

photon flux density

PSII:

photosystem II

V:

violaxanthin

Z:

zeaxanthin

ΔSn, ΔZ:

spectral absorbance change caused by light-scattering, violaxanthin deepoxidation

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Author notes

  1. Wolfgang Bilger

    Present address: Lehrstuhl für Botanik II der Universität, Mittlerer Dallenbergweg 64, D-97082, Würzburg, FRG

Authors and Affiliations

  1. Department of Plant Biology, Carnegie Institution of Washington, 290 Panama Street, 94305, Stanford, CA, USA

    Wolfgang Bilger & Olle Björkman

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  1. Wolfgang Bilger
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  2. Olle Björkman
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Additional information

We thank Connie Shih for skillful assistance in growing the plants, and for conducting HPLC analyses. A Carnegie Institution Fellowship and a Feodor-Lynen-Fellowship by the Alexander von Humboldt-Foundation to W. B. is gratefully acknowledged. This work was supported in part by Grant No. 89-37-280-4902 of the Competitive Grants Program of the U.S. Department of Agriculture to O.B. This is C. I. W. — D. P. B. Publication No. 1094.

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Bilger, W., Björkman, O. Relationships among violaxanthin deepoxidation, thylakoid membrane conformation, and nonphotochemical chlorophyll fluorescence quenching in leaves of cotton (Gossypium hirsutum L.). Planta 193, 238–246 (1994). https://doi.org/10.1007/BF00192536

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