Abstract
Two different preparations of photosystem II (PSII) (BBY-type membrane fragments and PSII core complexes) were isolated from 14-day-old pea seedlings (Pisum sativum L.) and used for spectral and kinetic study of photobleaching of chlorophyll (Chl) and amino acids under photoinhibitory conditions. A short-term (2–4 min) illumination of PSII preparations with high-intensity red light (λ > 610 nm, 800 W/m2) resulted in irreversible photobleaching of Chl at 672 and 682 nm under conditions of both acceptor- and donor-side photoinhibition. At longer illumination exposures (> 10 min) the photobleaching maximum at 682 nm was predominant. The calculated kinetic constants for Chl photobleaching in both absorption bands at temperatures of 20 and 4°C had similar values under different photoinhibitory conditions. The shape of action spectrum for Chl photooxidation indicates that photoinhibition of PSII was sensitized by two spectral forms of Chl with absorption maxima at 670 and 680 nm. The photobleaching of amino acids in PSII membrane fragments was only observed during acceptor-side photoinhibition and displayed the photobleaching peaks at 220 and 274 nm. The photogeneration of superoxide anion radical during donor-side photoinhibition was 4–6 times larger than during acceptor-side photoinhibition. Nevertheless, the kinetics of Chl and amino acid photobleaching in PSII preparations showed no appreciable differences. The activation energies for Chl photooxidation were estimated around 3.5 and 9 kcal/mol during acceptor- and donor-side photoinhibition, respectively, providing evidence for the involvement of biochemical stages in PSII photoinhibition. Based on the data obtained, it is proposed that the antenna Chl, rather than Chl of the reaction center, is the sensitizer for both acceptor- and donor-side photoinhibition of PSII in vitro.
Similar content being viewed by others
Abbreviations
- Car:
-
carotenoids
- Chl:
-
chlorophyll
- OEC:
-
oxygen-evolving complex
- Pheo:
-
pheophytin
- RC:
-
reaction center
- ROS:
-
reactive oxygen species
- k OX :
-
rate constant of Chl photooxidation
References
Powels, S.B., Photoinhibition of Photosynthesis Induced by Visible Light, Annu. Rev. Plant Physiol., 1985, vol. 35, pp. 15–44.
Aro, E.-M., Virgin, I., and Andersson, B., Photoinhibition of Photosystem II. Inactivation, Protein Damage and Turnover, Biochem. Biophys. Acta, 1993, vol. 1143, pp. 113–134.
Yamamoto, Y., Quality Control of Photosystem II, Plant Cell Physiol., 2001, vol. 42, pp. 121–128.
Klimov, V.V., Klevanik, A.V., Shuvalov, V.A., and Krasnovsky, A.A., Reduction of Pheophytin in the Primary Light Reaction of Photosystem II, FEBS Lett., 1977, vol. 82, pp. 183–186.
Vass, I., Styring, S., Hundall, T., Koivuniemi, A., Aro, E.-M., and Andersson, B., Reversible and Irreversible Intermediates during Photoinhibition of Photosystem II: Stable Reduced QA Species Promote Chlorophyll Triplet Formation, Proc. Natl. Acad. Sci. USA, 1992, vol. 89, pp. 1408–1412.
Zhang, L. and Aro, E.-M., Synthesis, Membrane Insertion and Assembly of the Chloroplast-Encoded D1 Protein into Photosystem II, FEBS Lett., 2002, vol. 512, pp. 13–18.
Mishra, N.P., Francko, C., von Gorkom, H.J., and Ghanotakis, D.F., Destructive Role of Singlet Oxygen during Aerobic Illumination of the Photosystem II Core Complex, Biochim. Biophys. Acta, 1994, vol. 1186, pp. 81–90.
Hideg, E., Spetea, C., and Vass, I., Singlet Oxygen and Free Radical Production during Acceptor- and Donor-Side-Induced Photoinhibition. Studies with Spin Trapping EPR Spectroscopy, Biochim. Biophys. Acta, 1994, vol. 1186, pp. 143–152.
Chen, G.X., Blubaugh, D.J., Homann, P.H., Goldbeck, J.H., and Cheniae, G.M., Superoxide Contributes to the Rapid Inactivation of Specific Secondary Donors of Photosystem Reaction Center during Photodamage of Manganese-Depleted PS II Membranes, Biochemistry, 1995, vol. 34, pp. 2317–2332.
Mizusawa, N., Tomo, T., Satoh, K., and Miyao, M., Degradation of the D1 Protein of Photosystem II under Illumination In Vivo: Two Different Pathways Involving Cleavage or Intermolecular Cross-Linking, Biochemistry, 2003, vol. 42, pp. 10 034–10 044.
Lupinkova, L. and Komenda, J., Oxidative Modifications of the Photosystem II D1-Protein by Reactive Oxygen Species: From Isolated Protein to Cyanobacterial Cells, Photochem. Photobiol., 2004, vol. 79, pp. 152–162.
Anderson, J.M., Park, Y.-I., and Chow, W.S., Unifying Model for the Photoinactivation of Photosystem II In Vivo under Steady-State Photosynthesis, Photosynth. Res., 1998, vol. 56, pp. 1–13.
Noguchi, T., Dual Role of Triplet Localization on the Accessory Chlorophyll in the Photosystem II Reaction Center: Photoprotection and Photodamage of the D1 Protein, Plant Cell Physiol., 2002, vol. 43, pp. 1112–1116.
Santabarbara, S., Cazzalini, I., Rivadossi, A., Garlaschi, F.M., Zucchelli, G., and Jenings, R.C., Photoinhibition In Vivo and In Vitro Involves Weakly Coupled Chlorophyll-Protein Complexes, Photochem. Photobiol., 2002, vol. 75, pp. 613–618.
Santabarbara, S., Neverov, K.V., Garlaschi, F.M., Zucchelli, G., and Jennings, R.C., Involvement of Uncoupled Antenna Chlorophylls in Photoinhibition in Thylakoids, FEBS Lett., 2001, vol. 491, pp. 109–113.
Neverov, K.V. and Krasnovsky, A.A., Jr., Phosphorescence Analysis of Chlorophyll Triplet State Formation in Preparations of Photosystem II, Biofizika, 2004, vol. 49, pp. 493–498.
Tjus, S.E., Scheller, H.V., Andersson, B., and Moller, B.L., Active Oxygen Produced during Selective Excitation of Photosystem I Is Damaging Not Only to Photosystem I, but Also to Photosystem II, Plant Physiol., 2001, vol. 125, pp. 2007–2015.
Strizh, I.G., Lysenko, G.G., and Neverov, K.V., Photoreduction of Molecular Oxygen in Preparations of Photosystem II under Photoinhibitory Conditions, Russ. J. Plant Physiol., 2005, vol. 52, pp. 717–723.
Berthold, D.A., Babcock, G.T., and Yocum, C.F., A High Resolved Oxygen-Evolving Photosystem II Preparation from Spinach Thylakoid Membranes, FEBS Lett., 1981, vol. 134, pp. 231–234.
Ghanotakis, D.F., Demetriou, D.M., and Yocum, C.F., Isolation and Characterization of an Oxygen Evolving Photosystem II Reaction Center Core Preparation and a 28 kDa Chl-a-Binding Protein, Biochim. Biophys. Acta, 1987, vol. 891, pp. 5–21.
Arnon, D.I., Copper Enzymes in Isolated Chloroplasts. Polyphenoloxidase in Beta vulgaris, Plant Physiol., 1949, vol. 24, pp. 1–15.
Klimov, V., Ananyev, G., Zastryzhnaya, O., Wydrzynski, T., and Renger, G., Photoproduction of Hydrogen Peroxide in Photosystem II Membrane Fragments: A Comparison of Four Signals, Photosynth. Res., 1993, vol. 38, pp. 409–416.
De Las Rivas, J., Telfer, A., and Barber, J., Two Coupled β-Carotene Molecules Protect P680 from Photodamage in Isolated Photosystem II Reaction Centres, Biochim. Biophys. Acta, 1993, vol. 1142, pp. 155–164.
Egorov, S.Yu., Krasnovsky, A.A., Jr., and Kulakovskaya, L.I., Investigations of Chloroplast Photodestruction Mechanism: Chlorophyll Triplet State Participation, Sov. Plant Physiol., 1985, vol. 32, pp. 668–673.
Krasnovsky, A.A., Jr., Singlet Molecular Oxygen: Mechanisms of Its Production and Deactivation in Photosynthetic Systems, Biofizika, 1994, vol. 39, pp. 236–250.
Krasnovsky, A.A., Jr., Singlet Oxygen in Photosynthetic Organisms, Zh. Vses. Khim. O-va im. D.I. Mendeleeva, 1986, vol. 31, pp. 562–568.
Neverov, K.V., Shalygo, N.V., Averina, N.G., and Krasnovsky, A.A., Jr., Pigment Triplet States in Green Leaves of Plants Treated with Metal-Chelating Agents, Russ. J. Plant Physiol., 1996, vol. 43, pp. 54–63.
Macpherson, A.N., Telfer, A., Barber, J., and Triscott, T.G., Direct Detection of Singlet Oxygen from Isolated Photosystem II Reaction Centres, Biochim. Biophys. Acta, 1993, vol. 1143, pp. 301–309.
Miller, R.W. and Macdowall, F.D.H., The Tiron Free Radical as a Sensitive Indicator of Chloroplastic Photoautoxidation, Biochim. Biophys. Acta, 1975, vol. 387, pp. 178–187.
Finkelstein, A.V. and Ptitsyn, O.B., Fizika belka (Physics of Proteins), Moscow: Knizhn. Dom Univ., 2002.
Baier, M. and Dietz, K.-J., Chloroplast as Source and Target of Cellular Redox Regulation: A Discussion on Chloroplast Redox Signals in the Context of Plant Physiology, J. Exp. Bot., 2005, vol. 416, pp. 1449–1462.
Author information
Authors and Affiliations
Additional information
Original Russian Text © I.G. Strizh, K.V. Neverov, 2007, published in Fiziologiya Rastenii, 2007, Vol. 54, No. 4, pp. 499–510.
Rights and permissions
About this article
Cite this article
Strizh, I.G., Neverov, K.V. Photoinhibition of photosystem II in vitro: Spectral and kinetic analyses. Russ J Plant Physiol 54, 439–449 (2007). https://doi.org/10.1134/S1021443707040024
Received:
Issue Date:
DOI: https://doi.org/10.1134/S1021443707040024