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The cyclic electron pathways around photosystem I in Chlamydomonas reinhardtii as determined in vivo by photoacoustic measurements of energy storage

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Abstract

The photoacoustic technique was used to measure energy storage by cyclic electron transfer around photosystem I in intact Chlamydomonas reinhardtii cells illuminated with far-red light (>715 nm). The in-vivo cyclic pathway was characterized by investigating the effects of various chemicals on energy storage. Participation of plastoquinone and ferredoxin in the cyclic electron flow was confirmed by the complete suppression of energy storage in the presence of the plastoquinol antagonist 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (DBMIB) and the ferredoxin inhibitors/competitors methylviologen, phenylmercuric acetate and p-benzoquinone. Two alternative electron cycles are demonstrated to operate in vivo. One cycle is sensitive to antimycin A, myxothiazol and 2-(n-heptyl)-4-hydroxyquinoline N-oxide (HQNO) and is catalyzed by ferredoxin which reduces plastoquinone through a route involving cytochrome b 6 and its protonmotive Q-cycle. The other cycle is unaffected by the above-mentioned inhibitors but is sensitive to N-ethylmaleimide (NEM), an inhibitor of the ferredoxin-NADP reductase, and 2′-monophosphoadenosine-5′-diphosphoribose (PADR), an analogue of NADP, showing that the electron recycling was mediated by NADPH. Possibly, electrons enter the plastoquinone pool through the action of a NAD(P)H dehydrogenase, which is insensitive to classical inhibitors of the mitochondrial NADH dehydrogenase. Loss of energy storage by photosystem-I-driven cyclic electron transfer in farred light was observed only when antimycin A, myxothiazol or HQNO was used in combination with NEM or PADR. Analysis of the light-intensity dependence and the rate of in-vivo cyclic electron transfer in the presence of various inhibitors indicates that the NADPH-dependent electron-cycle is the preferential cyclic pathway in Chlamydomonas cells illuminated with far-red light.

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Abbreviations

Amax :

maximal photothermal signal

Cyt:

cytochrome

DBMIB:

2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone

DCMU (diuron):

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

ES:

photochemical energy storage

FNR:

ferredoxin NADP+ reductase

HQNO:

2-(n-heptyl)-4-hydroxyquinoline N-oxide

NEM:

N-ethylmaleimide

P700 :

reaction-center pigment of PSI

PADR:

2′-monophosphoadenosine-5′-diphosphoribose

pBQ:

p-benzoquinone

PMA:

phenylmercuric acetate

References

  • Arnon, D.I., Chain, R.K. (1979) Regulatory electron transport pathways in cyclic photophosphorylation. FEBS Lett. 102, 133–138

    Google Scholar 

  • Asada, K., Heber, U., Schreiber, U. (1992) Pool size of electrons that can be donated to P700+, as determined in intact leaves: Donation to P700+ from stromal components via the intersystem chain. Plant Cell Physiol. 33, 927–932

    Google Scholar 

  • Avron, M. (1981) Photosynthetic electron transport and photophosphorylation. In: The biochemistry of plants, vol. 8, pp. 163–191, Hatch, M.D., Boardman, N.K., eds. Academic Press, New York

    Google Scholar 

  • Bendall, D.S. (1982) Photosynthetic cytochromes of oxygenic organisms. Biochim. Biophys. Acta 683, 119–151

    Google Scholar 

  • Bennoun, P. (1982) Evidence for a respiratory chain in the chloroplast. Proc. Natl. Acad. Sci. USA 79, 4352–4356

    Google Scholar 

  • Berger, S., Ellersiek, U., Westhoff, P., Steinmüller, K. (1993) Studies on the expression of NDH-H, a subunit of the NAD(P)H-plastoquinone-oxidoreductase of higher-plant chloroplasts. Planta 190, 25–31

    Google Scholar 

  • Bookjans, G., Böger, P. (1979) Complex-forming properties of butanedione-modified ferredoxin-NADP+ reductase with NADP+ and ferredoxin. Arch. Biochem. Biophys. 194, 387–393

    Google Scholar 

  • Brown, L.E., Sprecher, S.L., Keller, L.R. (1991) Introduction of exogenous DNA into Chlamydomonas reinhardtii by electroporation. Mol. Cell. Biol. 11, 2328–2332

    Google Scholar 

  • Bulls, G., Horwitz, B., Malkin, S., Cahen, D. (1982) Photoacoustic measurements of photosynthetic activities in whole leaves: photochemistry and gas exchange. Biochim. Biophys. Acta 679, 452–465

    Google Scholar 

  • Cahen, D. (1981) Photoacoustic cell for reflection and transmission measurements. Rev. Sci. Instrum. 52, 1306–1310

    Google Scholar 

  • Canaani, O., Schuster, G., Ohad, I. (1989) Photoinhibition in Chlamycomonas reinhardtii: Effect of state transition, intersystem energy distribution and photosystem I cyclic electron flow. Photosynth. Res. 20, 129–146

    Google Scholar 

  • Carpentier, R., Leblanc, R.M., Mimeault, M. (1990) On the nature of the photosynthetic energy storage monitored by photoacoustic spectroscopy. Photosynth. Res. 23, 313–318

    Google Scholar 

  • Cha, Y., Mauzerall, D.C. (1992) Energy storage of linear and cyclic electron flows in photosynthesis. Plant Physiol. 100, 1869–1877

    Google Scholar 

  • Chain, R.K., Arnon, D.I. (1977) Quantum efficiency of photosynthetic energy conversion. Proc. Natl. Acad. Sci. USA 74, 3377–3381

    Google Scholar 

  • Cleland, R.E., Bendall, D.S. (1992) Photosystem I cyclic electron transport: measurement of ferredoxin-plastoquinone reductase activity. Photosynth. Res. 34, 409–418

    Google Scholar 

  • Crofts, A.R., Wraight, C.A. (1983) The electrochemical domain of photosynthesis. Biochim. Biophys. Acta 726, 149–185

    Google Scholar 

  • Delieu, T., Walker, D.A. (1972) An improved cathode for the measurement of photosynthetic oxygen evolution by isolated chloroplasts. New Phytol. 71, 201–225

    Google Scholar 

  • dePamphilis, C.W., Palmer, J.D. (1990) Loss of photosynthetic and chlororespiratory genes from the plastid genome of a parasitic flowering plant. Nature 348, 337–339

    Google Scholar 

  • Diner, B., Mauzerall, D. (1973) Feedback controlling oxygen production in a cross-reaction between two photosystems in photosynthesis. Biochim. Biophys. Acta 305, 329–352

    Google Scholar 

  • Fork, D.C., Herbert, S.K. (1993) The application of photoacoustic techniques to studies of photosynthesis. Photochem. Photobiol. 57, 207–220

    Google Scholar 

  • Forti, G., Rosa, L. (1971) Cyclic photophosphorylation. In: Proceedings of the IInd international congress of photosynthetsis research, Stresa, pp. 1261–1270, Forti, G., Avron, M., Melandri, A., eds. Junk, The Hague

    Google Scholar 

  • Forti, G., Zanetti, G. (1969) The electron pathway of cyclic photophosphorylation. In: Progress in photosynthesis research, pp. 1213–1216, Metzner, H., ed. Loupp, Tübingen

    Google Scholar 

  • Greenbaum, N.L., Mauzerall, D.C. (1991) Effect of irradiance level on distribution of chlorophylls between PSII and PSI as determined from optical cross-sections. Biochim. Biophys. Acta 1057, 195–207

    Google Scholar 

  • Hauska, G., Hurt, E., Gabellini, N., Lockau, W. (1983) Comparative aspects of quinol-cytochrome c/plastocyanin oxidoreductases. Biochim. Biophys. Acta 726, 97–133

    Google Scholar 

  • Havaux, M. (1992) Photoacoustic measurements of cyclic electron flow around photosystem I in leaves adapted to light-states 1 and 2. Plant Cell Physiol. 33, 799–803

    Google Scholar 

  • Havaux, M., Lorrain, L., Leblanc, R.M. (1989) In vivo measurements of spectroscopic and photochemical properties of intact leaves using the ‘mirage effect’. FEBS Lett 250, 395–399

    Google Scholar 

  • Havaux, M., Greppin, H., Strasser, R.J. (1991) Functioning of photosystems I and II in pea leaves exposed to heat stress in the presence or absence of light. Planta 186, 88–98

    Google Scholar 

  • Heber, U., Egneus, H., Hanck, U., Jensen, M., Köster, S. (1978) Regulation of photosynthetic electron transport and photophosphorylation in intact chloroplasts and leaves of Spinacia oleracea L. Planta 143, 41–49

    Google Scholar 

  • Herbert, S.K., Fork, D.C., Malkin, S. (1990) Photoacoustic measurements in vivo of energy storage by cyclic electron flow in algae and higher plants. Plant Physiol. 94, 926–934

    Google Scholar 

  • Hind, G., Crowther, D., Shahak, Y., Slovacek, R.E. (1981) The function and mechanism of cyclic electron transport. In: Photosynthesis II. Electron transport and photophosphorylation, pp. 87–97, Akoyunoglou, G., ed. Balaban International Science Services, Philadelphia

    Google Scholar 

  • Honeycutt, R.C., Krogmann, D.W. (1972) Inhibition of chloroplast reactions with phenylmercuric acetate. Plant Physiol. 49, 376–380

    Google Scholar 

  • Hormann, H., Neubauer, C., Asada, K., Schreiber, U. (1993) Intact chloroplasts display pH 5 optimum of O2-reduction in the absence of methyl viologen: Indirect evidence for a regulatory role of superoxide protonation. Photosynth. Res. 37, 69–80

    Google Scholar 

  • Hosler, J.P., Yocum, C.F. (1985) Evidence for two cyclic photophosphorylation reactions concurrent with ferredoxin-catalyzed non-cyclic electron transport. Biochim. Biophys. Acta 808, 21–31

    Google Scholar 

  • Hosler, J.P., Yocum, C.F. (1987) Regulation of cyclic photophosphorylation during ferredoxin-mediated electron transport. Plant Physiol. 83, 965–969

    Google Scholar 

  • Inoue, Y., Ogawa, T., Shibata, K. (1973) Light-induced spectral changes of P700 in the 800-nm region in Anacystis and spinach lamellae. Biochim. Biophys. Acta 305, 483–487

    Google Scholar 

  • Jeanjean, R., Mathijs, H.C.P., Onana, B., Havaux, M., Joset, F. (1993) Exposure of the cyanobacterium Synechocystis PCC6803 to salt stress induces concerted changes in respiration and photosynthesis. Plant Cell Physiol. 34, 1073–1079

    Google Scholar 

  • Katona, E., Neimanis, S., Schönknecht, G., Heber, U. (1992) Photosystem I-dependent cyclic electron transport is important in controlling photosystem II activity in leaves under conditions of water stress. Photosynth. Res. 34, 449–464

    Google Scholar 

  • Knaff, D.B., Smith, J.M., Chain, R.K. (1980) Circular dichroism studies of ferredoxin: protein complexes. Arch. Biochem. Biophys. 199, 177–122

    Google Scholar 

  • Lam, E., Malkin, R. (1982) Ferredoxin-mediated reduction of cytochrome b-563 in a chloroplast cytochrome b-563/f complex. FEBS Lett. 141, 98–101

    Google Scholar 

  • Leegood, R.C., Crowther, D., Walker, D.A., Hind, G. (1983) Energetics of photosynthesis activities in Zea mays. I. Studies of the flash-induced electrochromic shift and fluorescence induction in bundle sheath cells. Biochim. Biophys. Acta 722, 116–126

    Google Scholar 

  • Malkin, S., Cahen, D. (1979) Photoacoustic spectroscopy and radiant energy conversion: theory of the effect with special emphasis on photosynthesis. Photochem. Photobiol. 29, 803–813

    Google Scholar 

  • Matsubayashi, T., Wakasugi, T., Shinozaki, K., Yamaguchi-Shinozaki, K., Zaita, N., Hidaka, T., Meng, B.Y., Ohto, C., Tanaka, M., Kato, A., Maruyama, T., Sugiura, M. (1987) Six chloroplast genes (ndhA-F) homologous to human mitochondrial genes encoding components of the respiratory chain NADH dehydrogenase are actively expressed: Determination of the splice sites in ndhA and ndhB pre-mRNAs. Mol. Gen. Genet. 210, 385–393

    Google Scholar 

  • Maxwell, P.C., Biggins, J. (1976) Role of cyclic electron transport in photosynthesis as measured by the photoinduced turnover of P700 in vivo. Biochem. 15, 3975–3981

    Google Scholar 

  • Mehler, A.H. (1951) Studies on reactions of illuminated chloroplasts. I. Mechanism of the reduction of oxygen and other Hill reagents. Arch. Biochem. Biophys. 33, 65–77

    Google Scholar 

  • Mi, H., Endo, T., Schreiber, U., Ogawa, T., Asada, K. (1992) Electron donation from cyclic and respiratory flows to the photosynthetic intersystem chain is mediated by pyridine nucleotide dehydrogenase in the cyanobacterium Synechocystis PCC 6803. Plant Cell Physiol. 33, 1233–1237

    Google Scholar 

  • Morehouse, K.M., Mason, R.P. (1988) The transition metal-mediated formation of the hydroxyl free radical during the reduction of molecular oxygen by ferredoxin-ferredoxin: NADP+ oxidoreductase. J. Biol. Chem. 263, 1204–1211

    Google Scholar 

  • Moss, D.A., Bendall, D.S. (1984) Cyclic electron transport in chloroplasts. The Q-cycle and the site of action of antimycin. Biochim. Biophys. Acta 767, 389–395

    Google Scholar 

  • Oettmeier, W., Masson, K., Soll, M. (1992) The acridones, new inhibitors of mitochondrial NADH: ubiquinone oxidoreductase (complex I). Biochim. Biophys. Acta 1099, 262–266

    Google Scholar 

  • Ogawa, T. (1991) A gene homologous to the subunit-2 gene of NADH dehydrogenase is essential to inorganic carbon transport of Synechocystis PCC6803. Proc. Natl. Acad. Sci. USA 88, 4275–4279

    Google Scholar 

  • Ogawa, T., Miyano, A., Inoue, Y. (1985) Photosystem-I-driven inorganic carbon transport in the cyanobacterium, Anacystis nidulans. Biochim. Biophys. Acta 808, 77–84

    Google Scholar 

  • Ohyama, K., Kohchi, T., Sano, T., Yamada, Y. (1988) Newly identified groups of genes in chloroplasts. Trends Biochem. Sci. 13, 19–22

    Google Scholar 

  • O'Keefe, D.P. (1983) Sites of cytochrome b-563 reduction, and the mode of action of DNP-INT and DBMIB in the chloroplast cytochrome b-563/f complex. FEBS Lett. 162, 349–354

    Google Scholar 

  • Peltier, G., Schmidt, G.W. (1991) Chlororespiration: an adaptation to nitrogen deficiency in Chlamydomonas reinhardtii. Proc. Natl. Acad. Sci. USA 88, 4791–4795

    Google Scholar 

  • Peltier, G., Thibault, P. (1983) Ammonia exchange and photorespiration in Chlamydomonas. Plant Physiol. 71, 888–892

    Google Scholar 

  • Ravenel, J., Peltier, G. (1991) Inhibition of chlororespiration by myxothiazol and antimycin A in Chlamydomonas reinhardtii. Photosynth. Res. 28, 141–148

    Google Scholar 

  • Rosencwaig, A. (1980) Photoacoustic spectroscopy. Annu. Rev. Biophys. Bioenerg. 9, 31–54

    Google Scholar 

  • Schreiber, U., Klughammer, C., Neubauer, C. (1988) Measuring P700 absorbance changes around 830 nm with a new type of pulse modulation system. Z. Naturforsch. 43c, 686–698

    Google Scholar 

  • Schürmann, P., Buchanan, B.B., Arnon, D.I. (1971) Role of cyclic photophosphorylation in photosynthetic carbon dioxide assimilation by isolated chloroplasts. Biochim. Biophys. Acta 267, 111–124

    Google Scholar 

  • Shahak, Y., Crowther, D., Hind, G. (1981) The involvement of ferredoxin-NADP+ reductase in cyclic electron transport in chloroplasts. Biochim. Biophys. Acta 636, 234–243

    Google Scholar 

  • Singer, T.P., Ramsay, R.P. (1992) NADH-ubiquinone oxidoreductase. In: Molecular mechanisms in bioenergetics, pp. 145–162, Ernster, L., ed. Elsevier, New York

    Google Scholar 

  • Slovacek, R.E., Crowther, D., Hind, G. (1980) Relative activities of linear and cyclic electron flows during chloroplast CO2-fixation. Biochim. Biophys. Acta 592, 495–505

    Google Scholar 

  • Tagawa, K., Tsujimoto, H.Y., Arnon, D.I. (1963) Role of chloroplast ferredoxin in the energy conversion process of photosynthesis. Proc. Natl. Acad. Sci. USA 49, 567–572

    Google Scholar 

  • Telfer, A., Barber, J. (1981) Redox reactions of cytochromes b-563 and f during cyclic electron flow induced by continuous illumination. In: Photosynthesis II. Electron transport and photophosphorylation, pp. 559–568, Akoyunoglou, G., ed. Balaban International Sciences Services, Philadelphia

    Google Scholar 

  • Trebst, A. (1974) Energy conservation in photosynthetic electron transport of chloroplasts. Annu. Rev. Plant Physiol. 25, 423–458

    Google Scholar 

  • Verméglio, A., Ravenel, J., Peltier, G. (1990) Chlororespiration: a respiratory activity in the thylakoid membrane of microalgae and higher plants. In: Recent advances in experimental phycology, vol. 7, pp. 188–205, Wiessner, W., Robinson, D.G., Starr, R.C., eds. Springer, Berlin

    Google Scholar 

  • Zanetti, G. (1976) A lysyl residue at the NADP binding site of ferredoxin-NADP reductase. Biochim. Biophys. Acta 445, 14–24

    Google Scholar 

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Authors and Affiliations

  1. Département de Physiologie Végétale et Ecosystèmes, CEA-Sciences du Vivant, Centre d'Etudes de Cadarache, F-13108, Saint-Paul-lez-Durance, France

    Jacques Ravenel, Gilles Peltier & Michel Havaux

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  1. Jacques Ravenel
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  2. Gilles Peltier
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  3. Michel Havaux
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We are very grateful to Dr. M.-H. Montane (Cadarache, Saint-Paul-lez-Durance, France) for her advice in the electroporation experiments.

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Ravenel, J., Peltier, G. & Havaux, M. The cyclic electron pathways around photosystem I in Chlamydomonas reinhardtii as determined in vivo by photoacoustic measurements of energy storage. Planta 193, 251–259 (1994). https://doi.org/10.1007/BF00192538

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