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Characterization of a nonphotochemical quenching-deficient Arabidopsis mutant possessing an intact PsbS protein, xanthophyll cycle and lumen acidification

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Abstract

Arabidopsis thaliana plants grown from ethyl methane sulfonate-treated seeds were screened for so-called que mutants, which are affected in non-photochemical energy quenching. Based on video imaging of chlorophyll fluorescence an energy dissipation mutant, que1, was identified, isolated and characterized. Similar to the npq mutants, the que1 mutant showed a drastically reduced capacity for pH-dependent energy dissipation, qE, but without affecting the Δ pH-dependent conformational changes at 535 nm (ΔA 535), which have been supposed to be obligatorily correlated with qE and to reflect pH-regulated binding of zeaxanthin to the PsbS protein. Western blot and DNA sequence analysis revealed that neither a reduced expression of the PsbS protein nor a mutation in the PsbS gene was responsible for the missing qE in que1. Measurements of 9-aminoacridine fluorescence quenching showed that the acidification of the thylakoid lumen was also not affected in the mutant. Furthermore, que1 was able to convert violaxanthin to zeaxanthin. However, unusual characteristics of zeaxanthin formation in the mutant pointed at an altered availability of violaxanthin for de-epoxidation. This was further accompanied by a decrease of the photochemical quenching of chlorophyll fluorescence (qP), an increase of the portion of oxidized P700 and a reduction of the electron transport rate. These characteristics indicate changes in the organization of the thylakoid membrane that affect linear electron transport (but not lumen acidification) and the formation of energy dissipation in photosystem II. Preliminary genetic analysis revealed that the phenotype of que1 is related to two different mutations, mapped to the lower arms of chromosomes 1 and 4.

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Abbreviations

Ax:

Antheraxanthin

Chl:

Chlorophyll

F V/F M :

Maximum quantum yield of photosystem II

LHC:

Light-harvesting complex

NPQ:

Non-photochemical quenching of chlorophyll fluorescence

PS I:

Photosystem I

PSII:

Photosystem II

PFD:

Photon flux density

qE:

Energy-dependent quenching of chlorophyll fluorescence

qI:

Photoinhibitory quenching of chlorophyll fluorescence

qP:

Photochemical quenching of chlorophyll fluorescence

Vx:

Violaxanthin

VxDE:

Violaxanthin deepoxidase

Zx:

Zeaxanthin

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Acknowledgements

We thank Dr. C. Funk (Stockholm, Sweden) for kindly providing us with antibodies against the PsbS protein and Dr. U. Heber (Würzburg, Germany) for his help in the measurements of spectral absorbance changes. This work was supported by the Deutsche Forschungsgemeinschaft (grant no. Ja 665/3)

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  1. Institute of Plant Biochemistry, Heinrich-Heine-University Düsseldorf, Universitätsstraße 1, 40225, Dusseldorf, Germany

    Ljudmila Kalituho, Thomas Graßes, Maria Graf, Jennifer Rech & Peter Jahns

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  1. Ljudmila Kalituho
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  2. Thomas Graßes
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  3. Maria Graf
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  4. Jennifer Rech
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  5. Peter Jahns
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Correspondence to Peter Jahns.

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Kalituho, L., Graßes, T., Graf, M. et al. Characterization of a nonphotochemical quenching-deficient Arabidopsis mutant possessing an intact PsbS protein, xanthophyll cycle and lumen acidification. Planta 223, 532–541 (2006). https://doi.org/10.1007/s00425-005-0093-z

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  • DOI: https://doi.org/10.1007/s00425-005-0093-z

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