Photosynthesis Research

, Volume 27, Issue 2, pp 97–108 | Cite as

Two sites of photoinhibition of the electron transfer in oxygen evolving and Tris-treated PS II membrane fragments from spinach

  • H. -J. Eckert
  • B. Geiken
  • J. Bernarding
  • A. Napiwotzki
  • H. -J. Eichler
  • G. Renger
Regular Papers


Photoinhibition was analyzed in O2-evolving and in Tris-treated PS II membrane fragments by measuring flash-induced absorption changes at 830 nm reflecting the transient P680+ formation and oxygen evolution. Irradiation by visible light affects the PS II electron transfer at two different sites: a) photoinhibition of site I eliminates the capability to perform a ‘stable’ charge separation between P680+ and QA- within the reaction center (RC) and b) photoinhibition of site II blocks the electron transfer from YZ to P680+. The quantum yield of site I photoinhibition (2–3×10-7 inhibited RC/quantum) is independent of the functional integrity of the water oxidizing system. In contrast, the quantum yield of photoinhibition at site II depends strongly on the oxygen evolution capacity. In O2-evolving samples, the quantum yield of site II photoinhibition is about 10-7 inhibited RC/quantum. After selective elimination of the O2-evolving capacity by Tris-treatment, the quantum yield of photoinhibition at site II depends on the light intensity. At low intensity (<3 W/m2), the quantum yield is 10-4 inhibited RC/quantum (about 1000 times higher than in oxygen evolving samples). Based on these results it is inferred that the dominating deleterious effect of photoinhibition cannot be ascribed to an unique target site or a single mechanism because it depends on different experimental conditions (e.g., light intensity) and the functional status of the PS II complex.

Key words

photoinhibition photosystem II P680+ reduction oxygen evolution 



absorption change at 830 nm


primary electron donor of PS II


photosystem II


2(N-morpholino)ethansulfonic acid


primary and secondary acceptors of PS II






fullwidth at half maximum




photon fluence rate




reaction center


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Allakhverdiev SI, Setlikova E, Klimov VV and Setlik I (1987) In photoinhibited photosystem II particles pheo-phytin photoreduction remains unimpaired. FEBS Lett 226: 186–190CrossRefGoogle Scholar
  2. Arntz B and Trebst A (1986) On the role of the QB-protein of PS II in photoinhibition. FEBS Lett 194: 43–49CrossRefGoogle Scholar
  3. Babcock GT and Sauer K (1975a) A rapid light induced transient in electron paramagnetic resonance signal II activated upon inhibition of photosynthetic oxygen evolution. Biochim Biophys Acta 376: 315–328PubMedGoogle Scholar
  4. Babcock GT and Sauer K (1975b) Two electron donation sites for exogenous reductants in chloroplast photosystem II. Biochim Biophys Acta 396: 48–62PubMedGoogle Scholar
  5. Babcock GT, Blankenship RE and Sauer K (1976) Reaction kinetics for positive charge accumulation on the water side of chloroplast photosystem II. FEBS Lett 61: 286–289CrossRefPubMedGoogle Scholar
  6. Berthold DA, Babcock GT and Yocum CA (1981) A highly resolved, oxygen-evolving photosystem II preparation from spinach thylakoid membranes. FEBS Lett 134: 231–234CrossRefGoogle Scholar
  7. Brettel K and Witt HT (1983) Reaction kinetics of the photo-oxidized chlorophyll a II in chloroplasts measured in the nanosecond range at 837 nm under repetitive flash exitation. Photobiochem Photobiophys 6: 253–260Google Scholar
  8. Callahan FE, Becker DW and Cheniae GM (1986) Studies on the photoactivation of the water-oxidizing enzyme. II. Characterization of weak light photoinhibition of PS II and its light-induced recovery. Plant Physiol 82: 261–269Google Scholar
  9. Cleland RE, Melis A and Neale PJ (1986) Mechanism of photoinhibition: photochemical reaction center inactivation in system II of chloroplasts. Photosynth Res 9: 79–88Google Scholar
  10. Cleland RE, Critchley C and Melis A (1987) Alteration of electron flow around P680: the effect on photoinhibition. In: Biggins J (ed) Progress in Photosynthesis Research, Vol 4, pp 27–30Google Scholar
  11. Conjeaud H and Mathis P (1980) The effect of pH on the reduction kinetics of P-680 in Tris-treated chloroplasts. Biochim Biophys Acta 590: 353–359PubMedGoogle Scholar
  12. Conjeaud H, Mathis P and Paillotin G (1979) Primary and secondary electron donors in photosystem II of chloroplasts. Biochim Biophys Acta 546: 280–291PubMedGoogle Scholar
  13. Critchley C (1988) The molecular mechanism of photoinhibition-facts and fiction. Aust J Plant Physiol 15: 27–41Google Scholar
  14. Dekker JP, Plijter JJ, Ouwehand L and vanGorkom HJ (1984) Kinetics of manganese redox transitions in the oxygen-evolving apparatus of photosynthesis. Biochim Biophys Acta 767: 176–179Google Scholar
  15. Demeter S, Neale P and Melis A (1987) Photoinhibition: impairment of the primar charge separation between P-680 and pheophytin in photosystem II of chloroplasts. FEBS Lett 214: 370–374CrossRefGoogle Scholar
  16. Eckert H-J (1982) Thesis, Technical University BerlinGoogle Scholar
  17. Eckert H-J, Renger G and Witt HT (1984) Reduction kinetics of the photo-oxidized chlorophyll a II +. FEBS Lett 167: 316–320CrossRefGoogle Scholar
  18. Eckert H-J, Renger G, Bernarding J, Faust P, Eichler H-J and Salk J (1987) Examination of fluorescence lifetime and radical-pair decay in photosystem II membrane fragments from spinach. Biochim Biophys Acta 893: 208–218Google Scholar
  19. Eckert H-J, Wydrzynski T and Renger G (1988) The effect of diisothiocyanostilbene-2,2′-disulfonic acid (DIDS), trifluo-perazine and lauroylcholinechloride on P-680+ reduction and oxygen evolution. Biochim Biophys Acta 932: 240–249Google Scholar
  20. Ford RC and Evans MCW (1985) Electron transfer to P-680+ in active and inhibited photosystem II fractions from higher plants. Biochim Biophys Acta 807: 1–9Google Scholar
  21. Geiken B, Eckert H-J, Bernarding J, Napiwotzki A and Renger G (1990) Effects of photoinhibition on the turnover of P680 in oxygen-evolving and Tris-treated PS II-membrane fragments from Spinach. In: Baltscheffsky M (ed) Current Research in Photosynthesis, Vol 2, pp 373–376. StockholmGoogle Scholar
  22. Gläser M, Wolff Ch and Renger G (1976) Indirect evidence for a very fast recovery of chlorophyll-a II in Spinach chloroplasts. Z Naturforsch 31c: 712–721Google Scholar
  23. Greenberg BM, Gaba V, Canaani O, Malkin S, Matoo AK and Edelman M (1989) Separate photosensitizers mediate degradation of the 32 kDa photosystem II reaction center protein in the visible and UV spectral regions. Proc Natl Acad Sci USA 86: 6617–6620PubMedGoogle Scholar
  24. Jones LW and Kok B (1966) Photoinhibition of chloroplast reactions. I. Kinetics and action spectra. Plant Physiol 41: 1037–43Google Scholar
  25. Krause GH, Köster S and Wong SC (1985) Photoinhibition of photosynthesis under unaerobic conditions studied in leaves and chloroplasts of Spinacia oleracea L. Planta (Berl) 165: 430–438Google Scholar
  26. Kyle DJ, Ohad I and Arntzen CJ (1984) Membrane protein damage and repair; selective loss of a quinone-protein function in chloroplast membranes. Proc Natl Acad Sci USA 81: 4070–4074Google Scholar
  27. Ley A and Mauzerall D (1982) Absolute absorption cross-sections for photosystem II and the minimum quantum requirement for photosynthesis in Chlorella vulgaris. Biochim Biophys Acta 680: 95–106Google Scholar
  28. Matoo AK, Hoffman-Falk H, Marder JB and Edelman M (1984) Regulation of protein metabolism: coupling of photosynthetic electron transport to in vivo degradation of the rapidly metabolised 32-kilodalton protein of the chloroplast membranes. Proc Natl Acad Sci USA 81: 1380–4Google Scholar
  29. Mäennpää P, Andersson B and Sundby C (1987) Difference in sensitivity to photoinhibition between photosystem II in the appressed and non-appressed thylakoid region. FEBS Lett 215: 31–36CrossRefGoogle Scholar
  30. Michel H and Deisenhofer J (1988) Relevance of the photosynthetic reaction center from purple bacteria to the structure of photosystem II. Biochemistry 27: 1–7Google Scholar
  31. Nedbal L, Masojidek J, Komenda J, Prasil O and Setlik I (1990) Three types of Photosystem II photoinactivation. 2. Slow processes. Photosynth Res 24: 89–97CrossRefGoogle Scholar
  32. Ohad I, Adir N, Koike H, Kyle D and Inoue Y (1990) Mechanism of photoinhibition in vivo. J Biol Chem 265: 1972–1974PubMedGoogle Scholar
  33. Powles SB (1984) Photoinhibition of photosynthesis induced by visible light. Annu Rev Plant Physiol 35: 15–44Google Scholar
  34. Reisman S and Ohad I (1986) Light-dependent degradation of the thylakoid 32-kDa QB-protein in isolated chloroplast membranes of Chlamydomonas reiinhardtii. Biochim Biophys Acta 849: 51–61Google Scholar
  35. Renger G and Wolff C (1976) The existence of a high photochemical turnover rate at the reaction centers of system II in Tris-washed chloroplasts. Biochim Biophys Acta 423: 610–614PubMedGoogle Scholar
  36. Renger G and Weiss W (1986a) Functional and structural aspects of photosynthetic water oxidation. Biochem Soc Trans 14: 17–20PubMedGoogle Scholar
  37. Renger G, Hagemann R and Fromme R (1986b) The susceptibility of the p-benzoquinone-mediated electron transport and atrazine binding to trypsin and its modification by CaCl2 in thylakoids and PS II membrane fragments. FEBS Lett 203: 210–214CrossRefGoogle Scholar
  38. Renger G, Völker M, Eckert H-J, Fromme R, Hohm-Veit S and Gräber P (1989) On the mechanism of photosystem II deterioration by UV-B irradiation. Photochem Photobiol 49: 97–105CrossRefGoogle Scholar
  39. Styring S, Virgin I, Ehrenberg A and Andersson B (1990) Strong light photoinhibition of electrontransport in photosystem II. Impairment of the function of the first quinone acceptor QA. Biochim Biophys Acta 1015: 269–278Google Scholar
  40. Theg SM, Filar LJ and Dilley RA (1986) Photoinactivation of chloroplasts already inhibited on the oxidizing side of photosystem II. Biochim Biophys Acta 849: 104–111Google Scholar
  41. Thompson LK and Brudvig GW (1988) Cytochrome b-559 may function to protect photosystem II from photoinhibition. Biochemistry 27: 6653–6658PubMedGoogle Scholar
  42. Thompson LK, Miller A-F, dePaula IC and Brudvig GW (1988) Electron donation in photosystem II. Israel J of Chem 28: 121–128Google Scholar
  43. Trebst A (1986) The topology of the plastoquinone and herbicide binding peptides of photosystem II in the thylakoid membrane. Z Naturforsch 41c: 240–245Google Scholar
  44. VanBest I and Mathis P (1978) Kinetics of the reduction of the oxidized primary electron donor of photosystem II in Spinach chloroplasts and in Chorella cells in the microsecond and nanosecond time ranges following flash excitation. Biochim Biophys Acta 503: 178–188PubMedGoogle Scholar
  45. VanMieghem F, Nitschke W, Mathis F and Rutherford AW (1989) The influence of the quinone-iron electron acceptor complex on the reaction centre photochemistry of photosystem II. Biochim Biophys Acta 977: 207–214Google Scholar
  46. Vass I, Mohanty N and Demeter S (1988) Photoinhibition of electron transport activity of photosystem II in isolated thylakoids studied by thermoluminescence and delayed luminescence. Z Naturforsch 43c: 99–103Google Scholar
  47. Völker M, Ono T, Inoue Y and Renger G (1985) Effect of trypsin on PS-II particles. Correlation between Hill-activity, Mn-abundance and peptide pattern. Biochim Biophys Acta 806: 25–34Google Scholar
  48. Völker M, Eckert H-J and Renger G (1987) Effects of trypsin and bivalent cations on P680+-reduction, fluorescence induction and oxygen evolution in photosystem II membrane fragments by trypsin and CaCl2. Biochem Biophys Acta 890: 66–76Google Scholar
  49. Weiss W and Renger G (1984) UV-spectral characterization in Tris-washed chloroplasts of the redox component Dl which functionally connects the reaction center with the water-oxidizing enzyme system Y in photosynthesis. FEBS Lett 169: 219–223CrossRefGoogle Scholar
  50. Weiss W and Renger G (1986) Studies on the nature of the water-oxidizing enzyme. II. On the functional connection between the reaction-center complex and the water-oxidizing enzyme system Y in photosystem II. Biochim Biophys Acta 850: 173–183Google Scholar
  51. Yamashita T and Butler WL (1968) Photooxidation by photosystem II of Tris-washed chloroplasts. Plant Physiol 43: 1978–1984PubMedGoogle Scholar
  52. Zhao J and Brand JJ (1988) Sequential events in the photoinhibition of Synechosystis under sodium stress. Arch Biochem Biophys 264: 657–664PubMedGoogle Scholar

Copyright information

© Kluwer Academic Publishers 1991

Authors and Affiliations

  • H. -J. Eckert
    • 1
  • B. Geiken
    • 1
  • J. Bernarding
    • 2
  • A. Napiwotzki
    • 2
  • H. -J. Eichler
    • 2
  • G. Renger
    • 1
  1. 1.Max-Volmer-Institut für Biophysikalische und Physikalische ChemieTechnische Universität Berlin1 Berlin 12Germany
  2. 2.Optisches InstitutTechnische Universität Berlin1 Berlin 12Germany

Personalised recommendations