Skip to main content
SpringerLink
Log in

Photoinhibition in Euglena gracilis: Involvement of reactive oxygen species

  • Published:
Planta Aims and scope Submit manuscript

Abstract

Photoinhibition of isolated Euglena gracilis thylakoids was characterised by a drastic decline in PSII photochemistry, chlorophyll-a fluorescence and an enhanced degradation of the 32-kDa protein. The process of protein degradation, as shown by studies of [14C] atrazine binding, was clearly slower than the other events. The activity of PSI was not affected. Decrease of electron-transport activity and loss of herbicide binding were prevented in the presence of various antioxidants and enzymes which protect against free radicals; however, the protection was not total. The strongest effect was observed by addition of dimethylsulfoxide, a potent hydroxyl-radical (OH*) quencher. Furthermore, combinations of various protective substances were even more effective in reducing photoinhibition. Different reactive oxygen species, including H2O2, superoxide radicals and OH* radicals were obviously involved in photoinhibition. These results were confirmed by the addition of potential OH*-radical-generating substances. Simultaneous enhancement of OH*-radical formation and photoinhibitory damage were observed in these cases. The involvement of this highly toxic species could be shown directly by a colorimetric test, thus enabling its light-mediated formation during photoinhibition to be quantified for the first time. In all, the data indicate that a site in PSII is the origin of radical formation involved in photoinhibition and that H2O2 is an important precursor in the formation of hydroxyl-radicals.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

Chl:

chlorophyll

DCMU:

3-(3,4-dichlorophenyl)-1,1-dimethylurea

DCPIP:

2,6-dichlorphenolindophenol

DMSO:

dimethyl sulfoxide

FM :

maximum fluorescence

FV :

variable fluorescence

Fecy:

ferricyanide

MSA:

methane sulfinic acid

MV:

1,1′ dimethyl-4,4′ bipyridylium dichloride

OH* :

hydroxyl radical

PBQ:

p-phenylbenzoquinone

PDA:

p-phenylenediamine

PPFD:

photosynthetic photon flux density

SOD:

superoxide dismutase

References

  • Arnon, D.J. (1949) Copper enzymes in isolated chloroplasts. Poly-oxidases in Beta vulgaris. Plant Physiol. 24, 1–15

    Google Scholar 

  • Arntz, B., Trebst, A. (1986) On the role of the QB protein of PSII in photoinhibition. FEBS Lett. 194, 43–49

    Google Scholar 

  • Asada, K., Takahashi, M. (1987) Production and scavenging of active oxygen in photosynthesis. In: Topics in photosynthesis, vol. 9, pp. 227–288, Kyle, D.J., Osmond, C.B., Arntzen, C.J., eds. Elsevier, Amsterdam

    Google Scholar 

  • Asada, K., Kiso, K., Yoshikawa, K. (1974) Univalent reduction of molecular oxygen by spinach chloroplasts on illumination. J. Biol. Chem. 249, 2175–2181

    Google Scholar 

  • Babbs, C.F., Pham, J.A., Coolbaugh, R.C. (1989) Lethal hydroxyl radical production in paraquat-treated plants. Plant. Physiol. 90, 1267–1270

    Google Scholar 

  • Barenyi, B., Krause, G. (1985) Inhibition of photosynthetic reactions by light. Planta 163, 218–226

    Google Scholar 

  • Blubaugh, D.J., Cheniae, G.M. (1990) Kinetics of photoinhibition in hydroxylamine-extracted photosystem II membranes: relevance to photoactivation and sites of electron donation. Biochemistry 29, 5109–5118

    Google Scholar 

  • Bradley, R.L., Long, K.M., Frasch, W.D. (1991) The involvement of photosystem II-generated H2O2 in photoinhibition. FEBS Lett. 286, 209–213

    Google Scholar 

  • Cadenas, E. (1989) Biochemistry of oxygen toxicity. Ann. Rev. Biochem. 58, 79–110

    Google Scholar 

  • Callahan, F.E., Becker, D.W., Cheniae, G.M. (1986) Studies on the photoactivation of the water-oxidizing enzyme. II. Characterization of weak light photoinhibition of PSII and its light-induced recovery. Plant Physiol. 82, 261–269

    Google Scholar 

  • Callahan, F.E., Ghirardi, M.L., Sopory, S.K., Mehta, A.M., Edelman, M., Mattoo, A.K. (1990) A novel metabolic form of the 32 kDa-D1 protein in the grana-localized reaction center of photosystem II. J. Biol. Chem 265, 15357–15360

    Google Scholar 

  • Chow, W.S., Osmond, C.B., Huang, L.K. (1989) Photosystem II function and herbicide binding sites during photoinhibition of spinach chloroplasts in-vivo and in-vitro. Photosynthesis Res. 21, 17–26

    Google Scholar 

  • Cleland, R.E. (1988) Molecular events of photoinhibitory inactivation in the reaction centre of photosystem II. Aust. J. Plant Physiol. 15, 135–150

    Google Scholar 

  • Cleland, R.E., Melis, A., Neale, P.J. (1986) Mechanism of photoinhibition: photochemical reaction centres inactivation in system II of chloroplasts. Photosynth. Res. 9, 79–88

    Google Scholar 

  • Cornic, C., Miginiac-Maslow, M. (1985) Photoinhibition of photosynthesis in broken chloroplasts as a function of electron transfer rates during light treatment. Plant Physiol. 78, 724–729

    Google Scholar 

  • Czapski, G. (1984) Reaction of OH. Methods Enzymol. 105, 209–219

    Google Scholar 

  • Demeter, S., Neale, P.J., Melis, A. (1987) Photoinhibition: impairment of primary charge separation between P-680 and pheophytin in photosystem II of chloroplasts. FEBS Lett. 214, 370–374

    Google Scholar 

  • Demmig-Adams, B. (1990) Carotenoids and photoprotection in plants: A role for the xanthophyll zeaxanthin. Biochim. Biophys. Acta 1020, 1–24

    Google Scholar 

  • Elstner, E.F., Frommeyer, D. (1978) Production of hydrogen peroxide by photosystem II of spinach chloroplast lamellae. FEBS Lett. 86, 143–146

    Google Scholar 

  • Foote, S., Shook, F.C., Abakerci, R.B. (1984) Characterization of singlet oxygen. Methods Enzymol. 105, 36–47

    Google Scholar 

  • Hundal, T., Aro, E.-M., Carlberg, L, Andersson, B. (1990) Restoration of light induced photosystem II inhibition without de novo protein synthesis. FEBS Lett. 267, 203–206

    Google Scholar 

  • Jansen, M.A.K., Shaaltiel, Y., Kazzes, D., Canaani, O., Malkin, S., Gressel, J. (1989) Increased tolerance to photoinhibitory light in paraquat-resistant Conyza-bonariensis. Measured by photoacoustic spectroscopy and 14(CO2)-fixation. Plant Physiol. 91, 1174–1178

    Google Scholar 

  • Jung, J., Kim, H.S. (1990) The chromophores as endogenous sensitizers involved in the photogeneration of singlet oxygen in spinach thylakoids. Photochem. Photobiol. 52, 1003–1009

    Google Scholar 

  • Kettunen, R., Tyystjärvi, E., Aro, E.-M. (1991) D1 protein degradation during photoinhibition of intact leaves. A modification of the D1 protein precedes degradation. FEBS Lett. 290, 153–156

    Google Scholar 

  • Krause, G.H., Cornic, G. (1987) CO2 and O2 interactions in photoinhibition. In: Topics in photosynthesis, vol. 9, pp. 169–196, Kyle, D.J., Osmond, C.B., Arntzen, C.J., eds. Elsevier, Amsterdam

    Google Scholar 

  • Krause, G.H., Kirk, M., Heber, U., Osmond, C.B. (1978) O2 dependent inhibition of photosynthetic capacity in intact isolated chloroplasts and isolated cells from spinach leaves illuminated in the absence of CO2. Planta 142, 229–233

    Google Scholar 

  • Krause, G.H., Somersalo, S., Zumbusch, E., Weyers, B., Laasch, H. (1990) On the mechanism of photoinhibition in chloroplasts. Relationship between changes in fluorescence and activity of photosystem II. J. Plant Physiol. 136, 472–479

    Google Scholar 

  • Kuznicki, L., Mikolajczyk, E. Walne, P.L. (1990) Photobehavior of euglenoid flagellates: theoretical and evolutionary perspectives. Crit. Rev. Plant Sci. 9, 343–369

    Google Scholar 

  • Kyle, D.J., Ohad, I., Arntzen, C.J. (1984) Membrane protein damage and repair: selective loss of a quinone-protein function in chloroplast membranes. Proc. Natl. Acad. Sci. USA 81, 4070–4074

    Google Scholar 

  • Kyle, D.J. (1987) The biochemical basis for photoinhibition of photosystem II. In: Topics in photosynthesis, vol. 9, pp. 197–226, Kyle, D.J., Osmond, C.B., Arntzen, C.J., eds. Elsevier, Amsterdam

    Google Scholar 

  • Leatherbarrow, R.L. (1987) Enzfitter — a non-linear regression data analysis programe for the IBM PC, A manual, Elsevier, Amsterdam

    Google Scholar 

  • Le Gouallec, J.-L., Cornic, G. (1988) Photoinhibition of photosynthesis in Elatostema repens. Plant. Physiol. Biochem. 26, 705–712

    Google Scholar 

  • Lichtenthaler, H.K. (1987) Chlorophylls and carotenoids: pigments of photosynthetic biomembranes. Methods Enzymol. 148, 350–382

    Google Scholar 

  • Mattoo, A.K., Hoffmann-Falk, I.H., Marder, J.B., 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, 1380–1384

    Google Scholar 

  • Nedbal, L., Setlikova, E., Masojidek, J., Setlik, I. (1986) The nature of photoinhibition in isolated thylakoids. Biochim. Biophys. Acta 848, 108–119

    Google Scholar 

  • Nedbal, L., Masojidek, J., Komenda, J., Prasil, O., Setlik, I. (1990) Three types of photosystem II photoinactivation. 2. Slow processes. Photosynthesis Res. 24, 89–97

    Google Scholar 

  • Ohad, I., Kyle, D.J., Arntzen, C.J. (1984) Membrane protein damage and repair: removal and replacement of inactivated 32-kilodalton polypeptide in chloroplast membranes. J. Cell Biol. 99, 481–485

    Google Scholar 

  • Ohad, I., Koike, H., Shochat, S., Inoue, Y. (1988) Changes in the properties of reaction center II during the initial stages of photoinhibition as revealed by thermoluminescence measurements. Biochim. Biophys. Acta 933, 288–298

    Google Scholar 

  • Ohad, I., Adir, A., Koike, H., Kyle, D.J., Inoue, Y. (1990) Mechanism of photoinhibition in vivo. A reversible light-induced conformational change of reaction center II is related to an irreversible modification of the D1 protein. J. Biol. Chem. 265, 1972–1979

    Google Scholar 

  • Price, C.A., Reardon, E.M. (1982) Isolation of chloroplast for protein synthesis from spinach and Euglena gracilis by centrifugation in silica gels. In: Methods in chloroplast molecular biology, pp. 189–209, Edelman, M., Hallick, R.B., Chua, N.H., eds. Elsevier, Amsterdam

    Google Scholar 

  • Richter, M., Rühle, W., Wild, A. (1990) Studies on the mechanism of photosystem II photoinhibition. II. The involvement of toxic oxygen species. Photosynt. Res 24, 237–243

    Google Scholar 

  • Robinson, J.M. (1988) Does O2 photoreduction occur within chloroplasts in vivo. Physiol. Plant. 72, 666–680

    Google Scholar 

  • Schuster, G., Timberg, R., Ohad, I. (1988) Turnover of thylakoid photosystem II proteins during photoinhibition of Chlamydomonas reinhardtii. Eur. J. Biochem. 177, 403–410

    Google Scholar 

  • Shigeoka, S., Yasumoto, R., Onishi, T., Nakano, Y., Kitaoka, S. (1987) Properties of monodehydroascorbate reductase and dehydroascorbate reductase and their participation in the regeneration of ascorbate in Eurlena gracilis. J. Gen. Microbiol. 133, 227–232

    Google Scholar 

  • Sopory, S.K., Greenberg, B.M., Mehta, R.A., Edelman, M., Mattoo, A.K. (1990) Free-radical scavengers inhibit light-dependent degradation of 32 kDa photosystem II reaction center protein. Z. Naturforsch. 45C, 412–417

    Google Scholar 

  • Theg, S.M., Filar, L.J., Dilley R.A. (1986) Photoinactivation of chloroplasts already inhibited on the oxidizing side of photosystem II. Biochim. Biophys. Acta 849, 104–111

    Google Scholar 

  • Tischer, W., Strotmann, H. (1977) Relationship between inhibitor binding by chloroplasts and inhibition of photosynthetic electron transport. Biochim. Biophys. Acta 460, 113–125

    Google Scholar 

  • Van Wijk, K.J., Krause, G.H. (1991) Oxygen dependence of photoinhibition at low temperature in intact protoplasts of Valerianella locusta L. Planta 186, 135–142

    Google Scholar 

  • Virgin, I., Styring, S., Andersson, B. (1988) Photosystem II disorganization and manganese release after photoinhibition of isolated spinach thylakoid membranes. FEBS Lett. 233, 408–412

    Google Scholar 

  • Wu, J., Neimanis, S., Heber, U. (1991) Photorespiration is more effective than the Mehler reaction in protecting the photosynthetic apparatus against photoinhibition. Bot. Acta 104, 283–291

    Google Scholar 

  • Wydrzynski, T., Ångström, J., Vänngård, T. (1989) H2O2 formation by photosystem II. Biochim, Biophys. Acta 973, 23–28

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Martin Luther Universität Halle-Wittenberg, Institut für Pflanzen- und Zellphysiologie, Am Kirchtor 1, D-06108, Halle/S, Germany

    H. Tschiersch & E. Ohmann

Authors
  1. H. Tschiersch
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. E. Ohmann
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

This reasearch was supported by a grant from the Volkswagen-Stiftung.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tschiersch, H., Ohmann, E. Photoinhibition in Euglena gracilis: Involvement of reactive oxygen species. Planta 191, 316–323 (1993). https://doi.org/10.1007/BF00195688

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00195688

Key words

Search

Navigation