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Elevated ΔpH restores rapidly reversible photoprotective energy dissipation in Arabidopsis chloroplasts deficient in lutein and xanthophyll cycle activity

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Abstract

The xanthophylls of the light-harvesting complexes of photosystem II (LHCII), zeaxanthin, and lutein are thought to be essential for non-photochemical quenching (NPQ). NPQ is a process of photoprotective energy dissipation in photosystem II (PSII). The major rapidly reversible component of NPQ, qE, is activated by the transmembrane proton gradient, and involves the quenching of antenna chlorophyll excited states by the xanthophylls lutein and zeaxanthin. Using diaminodurene (DAD), a mediator of cyclic electron flow around photosystem I, to enhance ΔpH we demonstrate that qE can still be formed in the absence of lutein and light-induced formation of zeaxanthin in chloroplasts derived from the normally qE-deficient lut2npq1 mutant of Arabidopsis. The qE induced by high ΔpH in lut2npq1 chloroplasts quenched the level of fluorescence when all PSII reaction centers were in the open state (F o state), protected PSII reaction centers from photoinhibition, was sensitive to the uncoupler nigericin, and was accompanied by absorption changes in the 410–565 nm region. Titrations show the ΔpH threshold for activation of qE in lut2npq1 chloroplasts lies outside the normal physiological range and is highly cooperative. Comparison of quenching in isolated trimeric (LHCII) and monomeric (CP26) light-harvesting complexes from lut2npq1 plants revealed a similarly shifted pH dependency compared with wild-type LHCII. The implications for the roles of lutein and zeaxanthin as direct quenchers of excitation energy are discussed. Furthermore, we argue that the control over the proton-antenna association constant, pK, occurs via influence of xanthophyll structure on the interconnected phenomena of light-harvesting antenna reorganization/aggregation and hydrophobicity.

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Abbreviations

PSI:

Photosystem I

PSII:

Photosystem II

LHCII:

Light-harvesting antenna complex of photosystem II

CP26:

Minor PSII light-harvesting complex

ΔpH:

Proton gradient across the thylakoid membrane

NPQ:

Nonphotochemical chlorophyll fluorescence quenching

qE:

ΔpH-dependent portion of NPQ

(z):

Wild-type zeaxanthin containing chloroplasts

(v):

Wild-type chloroplasts containing no zeaxanthin

pK :

Proton antenna association constant

LEDs:

Light-emitting diodes

F o :

Maximum chlorophyll fluorescence in when all PSII reaction centers are open, 9-aa, 9-aminoacridine

DAD:

Diaminodurene

CEF:

Cyclic electron flow

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Acknowledgments

This work was supported by research and equipment grants to A.V.R. from the Royal Society and UK Biotechnology and Biological Sciences Research Council and Engineering and Physical Sciences Research Council. The authors wish to thank Krishna Niyogi (University of California, Berkeley, USA) for providing the seeds of the Arabidopsis mutant used in this study.

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  1. School of Biological and Chemical Sciences, Queen Mary University of London, Mile End Road, Fogg Building, London, E1 4NS, UK

    Matthew P. Johnson, Ahmad Zia & Alexander V. Ruban

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  1. Matthew P. Johnson
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  2. Ahmad Zia
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  3. Alexander V. Ruban
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Correspondence to Alexander V. Ruban.

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Johnson, M.P., Zia, A. & Ruban, A.V. Elevated ΔpH restores rapidly reversible photoprotective energy dissipation in Arabidopsis chloroplasts deficient in lutein and xanthophyll cycle activity. Planta 235, 193–204 (2012). https://doi.org/10.1007/s00425-011-1502-0

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