Photosynthesis Research

, Volume 94, Issue 2–3, pp 217–224 | Cite as

Application of low temperatures during photoinhibition allows characterization of individual steps in photodamage and the repair of photosystem II

  • Prasanna Mohanty
  • Suleyman I. Allakhverdiev
  • Norio MurataEmail author
Research Article


Recent investigations of photoinhibition have revealed that photodamage to photosystem II (PSII) involves two temporally separated steps: the first is the inactivation of the oxygen-evolving complex by light that has been absorbed by the manganese cluster and the second is the impairment of the photochemical reaction center by light that has been absorbed by chlorophyll. Our studies of photoinhibition in Synechocystis sp. PCC 6803 at various temperatures demonstrated that the first step in photodamage is not completed at low temperatures, such as 10°C. Further investigations suggested that an intermediate state, which is stabilized at low temperatures, might exist at the first stage of photodamage. The repair of PSII involves many steps, including degradation and removal of the D1 protein, synthesis de novo of the precursor to the D1 protein, assembly of the PSII complex, and processing of the precursor to the D1 protein. Detailed analysis of photodamage and repair at various temperatures has demonstrated that, among these steps, only the synthesis of the precursor to D1 appears to proceed at low temperatures. Investigations of photoinhibition at low temperatures have also indicated that prolonged exposure of cyanobacterial cells or plant leaves to strong light diminishes their ability to repair PSII. Such non-repairable photoinhibition is caused by inhibition of the processing of the precursor to the D1 protein after prolonged illumination with strong light at low temperatures.


D1 protein Low temperature Photodamage Photoinhibition Photosynthesis Photosystem II Processing Repair 



Dichlorophenol indophenol


Diphenyl carbazide


Oxygen-evolving complex


Precursor to the D1 protein


Reaction center


Reactive oxygen species



The authors acknowledge Dr. J. S. S. Prakash, Hyderabad University, and Dr. Shunichi Takahashi, the Australian National University, for their kind help during the preparation of the manuscript. This work was supported, in part, by the Cooperative Research Program on Stress-Tolerant Plants of the National Institute for Basic Biology, Japan, and by grants from the Russian Foundation for Basic Research (no. 05-04-49672) and from the Molecular and Cell Biology Programs of the Russian Academy of Sciences (to S.I.A.). P.M. thanks the Indian National Science Academy, New Delhi, for his assignment as an honorary scientist at RPRC Bhubaneswar.


  1. Adir N, Zer H, Shochat S, Ohad I (2003) Photoinhibition: a historical perspective. Photosynth Res 76:343–370PubMedCrossRefGoogle Scholar
  2. Allakhverdiev SI, Murata N (2004) Environmental stress inhibits the synthesis de novo of proteins involved in the photodamage-repair cycle of photosystem II in Synechocystis sp. PCC 6803. Biochim Biophys Acta 1657:23–32PubMedCrossRefGoogle Scholar
  3. Allakhverdiev SI, Nishiyama Y, Miyairi S, Yamamoto H, Inagaki N, Kanesaki Y, Murata N (2002) Salt stress inhibits the repair of photodamaged photosystem II by suppressing the transcription and translation of psbA genes in Synechocystis. Plant Physiol 130:1443–1453PubMedCrossRefGoogle Scholar
  4. Allakhverdiev SI, Mohanty P, Murata N (2003) Dissection of photodamage at low temperature and repair in darkness suggests the existence of an intermediate form of photodamaged photosystem II. Biochemistry 42:14277–14283PubMedCrossRefGoogle Scholar
  5. Allakhverdiev SI, Nishiyama Y, Takahashi S, Miyairi S, Suzuki I, Murata N (2005a) Systematic analysis of the relation of electron transport and ATP synthesis to the photodamage and repair of photosystem II in Synechocystis. Plant Physiol 137:263–273PubMedCrossRefGoogle Scholar
  6. Allakhverdiev SI, Tsvetkova N, Mohanty P, Szalontai, Moon BY, Debreczeny M, Murata N (2005b) Irreversible photoinhibition of photosystem II is caused by exposure of Synechocystis cells to strong light for a prolonged period. Biochim Biophys Acta 1708:342–351PubMedCrossRefGoogle Scholar
  7. Andersson B, Aro E-M (2001) Photodamage and D1 protein turnover in photosystem II. In: Aro E-M, Andersson B (eds) Regulation of photosynthesis. Kluwer Academic Publishers, Dordrecht, The Netherlands, pp 377–393Google Scholar
  8. Aro E-M, Virgin I, Andersson B (1993) Photoinhibition of photosystem II: inactivation, protein damage and turnover. Biochim Biophys Acta 1143:113–134PubMedCrossRefGoogle Scholar
  9. Baffert C, Collomb MN, Deronzier A, Pecaut J, Limburg J, Crabtree RH, Brudvig G (2002) Two new terpyridine dimanganese complexes: a manganese(III,III) complex with a single unsupported oxo bridge and a manganese(III,IV) complex with a dioxo bridge. Synthesis, structure, and redox properties. Inorg Chem 41:1404–1411PubMedCrossRefGoogle Scholar
  10. Barber J, Andersson B (1992) Too much of a good thing: light can be bad for photosynthesis. Trends Biochem Sci 17:61–66PubMedCrossRefGoogle Scholar
  11. Barker M, de Vries R, Nield J, Komenda J, Nixon PJ (2006) The Deg proteases protect Synechocystis sp. PCC 6803 during heat and light stresses but are not essential for removal of damaged D1 protein during the photosystem two repair cycle. J Biol Chem 281:30347–30355PubMedCrossRefGoogle Scholar
  12. Carrell TG, Bourles E, Lin M, Dismukes GC (2003) Transition from hydrogen atom to hydride abstraction by Mn4O4(O2PPh2)6 versus [Mn4O4(O2PPh2)6]+: O–H bond dissociation energies and the formation of Mn4O3(OH)(O2PPh2)6. Inorg Chem 42:2849–2858PubMedCrossRefGoogle Scholar
  13. Chow WS, Lee HY, He J, Hindrickson L, Hong YN, Matsubara S (2005) Photoinhibition in leaves. Photosynth Res 84:35–41PubMedCrossRefGoogle Scholar
  14. Constant S, Eisenberg-Domovitch Y, Ohad I, Kirilovsky D (2000) Recovery of photosystem II activity in photoinhibited Synechocystis cells: light-dependent translation activity is required besides light-independent synthesis of the D1 protein. Biochemistry 79:2032–2041CrossRefGoogle Scholar
  15. Erickson JM (1998) Assembly of photosystem II. In: Rochaix J-D, Goldschmidt-Clermont M, Merchant S (eds) The molecular biology of chloroplasts and mitochondria in Chlamydomonas. Kluwer Academic Publishers, Dordrecht, The Netherlands, pp 255–285Google Scholar
  16. Giacometti GM, Barbato R, Chiaramonte S, Friso G, Rigoni F (1996) Effects of ultraviolet-B radiation on photosystem II of the cyanobacterium Synechocystis sp. PCC 6803. Eur J Biochem 242:799–806PubMedCrossRefGoogle Scholar
  17. Hakala M, Tuominen I, Keränen M, Tyystjärvi T, Tyystjärvi E (2005) Evidence for the role of the oxygen-evolving manganese complex in photoinhibition of photosystem II. Biochim Biophys Acta 1706:68–80PubMedCrossRefGoogle Scholar
  18. Hideg E, Murata N (1997) The irreversible photoinhibition of the photosystem II complex in leaves of Vicia faba under strong light. Plant Sci 130:151–158CrossRefGoogle Scholar
  19. Hideg E, Spetea C, Vass I (1994) Singlet oxygen and free radical production during acceptor- and donor-side-induced photoinhibition. Studies with spin trapping EPR spectroscopy. Biochim Biophys Acta 1186:143–152CrossRefGoogle Scholar
  20. Huesgen PF, Schuhmann H, Adamska I (2006) Photodamaged D1 protein is degraded in Arabidopsis mutants lacking the Deg2 protease. FEBS Lett 580:6929–6932PubMedCrossRefGoogle Scholar
  21. Hundal T, Aro E-M, Carlberg I, Andersson B (1990) Restoration of light-induced photosystem II inhibition without de novo protein synthesis. FEBS Lett 267:203–206PubMedCrossRefGoogle Scholar
  22. Inagaki N, Yamamoto Y, Satoh K (2001) A sequential two-step proteolytic process in the carboxy-terminal truncation of precursor D1 protein in Synechocystis 6803. FEBS Lett 509:197–201PubMedCrossRefGoogle Scholar
  23. Jegerschöld C, Virgin I, Styring S (1990) Light-dependent degradation of the D1 protein in photosystem II is accelerated after inhibition of the water-splitting reaction. Biochemistry 29:6179–6186PubMedCrossRefGoogle Scholar
  24. Jones LW, Kok B (1966) Photoinhibition of chloroplast reactions. I. Kinetics and action spectra. Plant Physiol 41:1037–1043PubMedCrossRefGoogle Scholar
  25. Jung J, Kim HS (1990) The chromophores as endogenous sensitizers involved in the photogeneration of singlet oxygen in spinach thylakoids. Photochem Photobiol 52:1003–1009Google Scholar
  26. Kanervo E, Aro E-M, Murata N (1995) Low unsaturation level of thylakoid membrane lipids limits turnover of the Dl protein of photosystem II at high irradiance. FEBS Lett 364:239–242PubMedCrossRefGoogle Scholar
  27. Kanervo E, Tasaka Y, Murata N, Aro A-M (1997) Membrane lipid unsaturation modulates processing of the photosystem II reaction-center protein D1 at low temperature. Plant Physiol 114:841–849PubMedCrossRefGoogle Scholar
  28. Keren N, Ohad I (1998) State transition and photoinhibition. In: Rochaix J-D, Goldschmidt-Clermont M, Merchant S (eds) The molecular biology of chloroplasts and mitochondria in Chlamydomonas. Kluwer Academic Publishers, Dordrecht, The Netherlands, pp 570–596Google Scholar
  29. Kirilovsky DL, Vernotte C, Etienne AL (1990) Protection from photoinhibition by low temperature in Synechocystis 6714 and in Chlamydomonas reinhardtii: detection of an intermediary state. Biochemistry 29:8100–8106PubMedCrossRefGoogle Scholar
  30. Kuvikova S, Tichy M, Komenda J (2005) A role of the C terminal extension of photosystem II D1 protein is sensitive of the cyanobacterium Synechocystis PCC 6803 to photoinhibition. Photochem Photobiol Sci 4:1044–1048PubMedCrossRefGoogle Scholar
  31. Kyle DJ, Ohad I, Arntzen CJ (1984) Membrane protein damage and repair: selective loss of a quinone-protein function in chloroplast membranes. Proc Natl Acad Sci USA 81:4070–4074PubMedCrossRefGoogle Scholar
  32. Lee HY, Hong YN, Chow WS (2001) Photoinactivation of photosystem II complexes and photoprotection by non-functional neighbors in Capsicum annuum L. leaves. Planta 212:332–342PubMedCrossRefGoogle Scholar
  33. Mattoo AK, Hoffman-Falk H, Marder JB, Edelman M (1984) Regulation of protein metabolism: coupling of photosynthetic electron transport to in vivo degradation of the rapidly metabolized 32-kilodalton protein of the chloroplast membranes. Proc Natl Acad Sci USA 81:1380–1384PubMedCrossRefGoogle Scholar
  34. Mattoo AK, Marder JB, Edelman M (1989) Dynamics of the photosystem II reaction center. Cell 56:241–246PubMedCrossRefGoogle Scholar
  35. Melis A (1999) Photosystem-II damage and repair cycle in chloroplasts: what modulates the rate of photodamage in vivo? Trends Plant Sci 4:130–135PubMedCrossRefGoogle Scholar
  36. Miyao M, Ikeuchi M, Yamamoto N, Ono T (1995) Specific degradation of the D1 protein of photosystem II by treatment with hydrogen peroxide in darkness: implication for the mechanism of degradation of the D1 protein under illumination. Biochemistry 34:10019–10026PubMedCrossRefGoogle Scholar
  37. Murata N, Takahashi S, Nishiyama Y, Allakhverdiev SI (2007) Photoinhibition of photosystem II under environmental stress. Biochim Biophys Acta (in press)Google Scholar
  38. Nishiyama Y, Yamamoto H, Allakhverdiev SI, Inaba M, Yokota A, Murata N (2001) Oxidative stress inhibits the repair of photodamage to the photosynthetic machinery. EMBO J 20:5587–5594PubMedCrossRefGoogle Scholar
  39. Nishiyama Y, Allakhverdiev SI, Yamamoto H, Hayashi H, Murata N (2004) Singlet oxygen inhibits the repair of photosystem II by suppressing the translation elongation of the D1 protein in Synechocystis sp. PCC 6803. Biochemistry 43:11321–11330PubMedCrossRefGoogle Scholar
  40. Nishiyama Y, Allakhverdiev SI, Murata N (2005) Inhibition of the repair of photosystem II by oxidative stress in cyanobacteria. Photosynth Res 84:1–7PubMedCrossRefGoogle Scholar
  41. Nishiyama Y, Allakhverdiev SI, Murata N (2006) A new paradigm for the action of reactive oxygen species in the photoinhibition of photosystem II. Biochim Biophys Acta 1757:742–749PubMedCrossRefGoogle Scholar
  42. Nixon P, Barker M, Boehm M, De Vries R, Komenda J (2005) FtsH-mediated repair of the photosystem II complex in response to light stress. J Exp Bot 56:357–363PubMedCrossRefGoogle Scholar
  43. Ohad I, Kyle DJ, Arntzen CJ (1984) Membrane protein damage and repair: removal and replacement of inactivated 32-kilodalton polypeptide in chloroplast membranes. J Cell Biol 99:481–485PubMedCrossRefGoogle Scholar
  44. Ohad I, Kyle DJ, Hirschberg J (1985) Light-dependent degradation of the QB-protein in isolated pea thylakoids. EMBO J 4:1655–1659PubMedGoogle Scholar
  45. Ohnishi N, Allakhverdiev SI, Takahashi S, Higashi S, Watanabe M, Nishiyama Y, Murata N (2005) Two-step mechanism of photodamage to photosystem II: step 1 occurs at the oxygen-evolving complex and step 2 occurs at the photochemical reaction center. Biochemistry 44:8494–8499PubMedCrossRefGoogle Scholar
  46. Öquist G, Huner NPA (2003) Photosynthesis of overwintering evergreen plants. Annu Rev Plant Biol 54:329–355PubMedCrossRefGoogle Scholar
  47. Ossenbuhi F, Inaba-Sulpice M, Meurer J, Soll J, Eichacker LA (2006) The Synechocystis PCC 6803 Oxa1 homolog is essential for integration of reaction center protein pD1. Plant Cell 18:2236–2246CrossRefGoogle Scholar
  48. Renger G, Volker M, Eckert HJ, Fromme R, Hohm-Veit S, Graber P (1989) On the mechanism of photosystem II deterioration by UV-B irradiation. Photochem Photobiol 49:97–105CrossRefGoogle Scholar
  49. Sippola K, Kanervo E, Murata N, Aro E-M (1998) A genetically engineered increase in fatty acid unsaturation in Synechococcus sp. PCC 7942 allows exchange of D1 protein forms and sustenance of photosystem II activity at low temperature. Eur J Biochem 251:641–648PubMedCrossRefGoogle Scholar
  50. Sundby C, Schioett T (1992) Characterization of the reversible state of photoinhibition occurring in vitro under anaerobic conditions. Photosynth Res 33:195–202CrossRefGoogle Scholar
  51. Szalontai B, Nishiyama Y, Gombos Z, Murata N (2000) Membrane dynamics as seen by Fourier transform infrared spectroscopy in a cyanobacterium, Synechocystis PCC 6803. The effects of lipid unsaturation and the protein-to-lipid ratio. Biochim Biophys Acta 1509:409–419PubMedCrossRefGoogle Scholar
  52. Tasaka Y, Gombos Z, Nishiyama Y, Mohanty P, Ohba T, Ohki K, Murata N (1996) Targeted mutagenesis of acyl-lipid desaturases in Synechocystis: evidence for the important roles of polyunsaturated membrane lipids in growth, respiration and photosynthesis. EMBO J 15:6416–6425PubMedGoogle Scholar
  53. Telfer A, Bishop SM, Phillips D, Barber J (1994) The isolated photosynthetic reaction center of PSII as a sensitiser for the formation of singlet oxygen; detection and quantum yield determination using a chemical trapping technique. J Biol Chem 269:13244–13253PubMedGoogle Scholar
  54. Tyystjärvi E, Aro E-M (1996) The rate constant of photoinhibition, measured in lincomycin-treated leaves, is directly proportional to light intensity. Proc Natl Acad Sci USA 93:2213–2218PubMedCrossRefGoogle Scholar
  55. Tyystjärvi T, Tuominen I, Herranen M, Aro E-M, Tyystjärvi E (2002) Action spectrum of psbA gene transcription is similar to that of photoinhibition in Synechocystis PCC 6803. FEBS Lett 516:167–171PubMedCrossRefGoogle Scholar
  56. Vass I, Styring S, Hundal T, Koivuniemi A, Aro E-M, Andersson B (1992) The reversible and irreversible intermediates during photoinhibition of photosystem II: stable reduced QA species promote chlorophyll triplet formation. Proc Natl Acad Sci USA 89:1408–1412PubMedCrossRefGoogle Scholar
  57. Zsiros O, Allakhverdiev SI, Higashi S, Watanabe M, Nishiyama Y, Murata N (2006) Very strong UV-A light temporally separates the photoinhibition of photosystem II into light-induced inactivation and repair. Biochim Biophys Acta 1757:123–129PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • Prasanna Mohanty
    • 1
    • 2
  • Suleyman I. Allakhverdiev
    • 3
  • Norio Murata
    • 4
    Email author
  1. 1.Jawaharlal Nehru UniversityNew DelhiIndia
  2. 2.Regional Plant Resource CenterNayapalli, BhubaneswarIndia
  3. 3.Institute of Basic Biological ProblemsRussian Academy of SciencesPushchino, Moscow RegionRussia
  4. 4.National Institute for Basic BiologyOkazakiJapan

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