, Volume 191, Issue 3, pp 365–376 | Cite as

Inhibition of photosynthesis, acidification and stimulation of zeaxanthin formation in leaves by sulfur dioxide and reversal of these effects

  • Sonja Veljovic-Jovanovic
  • Wolfgang Bilger
  • Ulrich Heber


Leaves of Pelargonium zonale L. and Spinacia oleracea L. were fumigated with high concentrations of SO2 for very short periods of time with the aim of first producing acute symptoms of damage and then observing repair. The response of different photosynthetic parameters to SO2 was monitored during and after fumigation. The following results were obtained: (1) Inhibition of CO2 assimilation in the light was accompanied by increased reduction of the quinone acceptor, QA, of photosystem II and by increased oxidation of the electrondonor pigment P700 of photosystem I. Increased control of photosystem II activity in the SO2-inhibited state was also indicated by increased light scattering and by increased non-photochemical quenching of chlorophyll fluorescence. Both are indicators of chloroplast energization. Apparently, SO2 did not decrease but rather increased energization of the chloroplast thylakoid system by light. (2) Accumulation of dihydroxyacetone phosphate, fructose-1,6-phosphate and ribulose-1,5-phosphate and a decrease of 3-phosphoglycerate and hexosephosphate indicated that SO2 inhibited enzymes of the Calvin cycle. (3) Stimulated postillumination CO2 evolution suggested that when photosynthesis declined respiration increased to provide energy for repair reactions. (4) Increased leaf absorbance at 505 nm indicated increased stimulation of zeaxanthin formation in thylakoid membranes under the influence of SO2. A similar increase in 505-nm absorbance could be induced by high concentrations of CO2. In darkened leaves, SO2 did not produce changes in 505-nm absorbance. (5) While zeaxanthin formation was stimulated, changes in the fluorescence of the pH-indicating dye pyranine, which had been fed to the leaves, indicated acidification of the cytoplasm of leaf cells by SO2. Maximum acid production by SO2 required light. In contrast, cytoplasmic acidification of leaf cells by CO2 was similar in the light and in the dark. (6) Since zeaxanthin formation is known to depend on the acidification of the thylakoid lumen, SO2-dependent zeaxanthin formation indicated SO2-dependent acidification of the thylakoid lumen as the indirect result of cytoplasmic acidification by SO2. (7) Inhibition of photosynthesis and other effects of SO2 were fully reversible in the light. Detoxification of SO2 and reactivation of the photosynthetic apparatus were slow or absent in the dark. Light had a dual effect on the action of SO2. Transiently, it first increased the extent of inhibition of assimilation, but, finally, it reversed inhibition. Sulfur dioxide was inhibitory as a consequence of the chemical reactivity of its hydration products rather than as a result of cellular acidification by the produced acid. The initial acidification was followed by an appreciable alkalisation demonstrating the action of the pH-stat mechanism. (8) The data are discussed in relation to SO2 toxicity under field conditions when plants are chronically exposed to polluted air.

Key words

Acidification Chlorophyll fluorescence Photosynthesis (SO2 effect) Xanthophyll cycle Spinacia Pelargonium 





dihydroxyacetone phosphate





F, Fm, Fm′, Fo, Fo′

chlorophyll fluorescence levels




primary donor of photosystem I


primary quinone acceptor of photosystem II


photochemical quenching of chlorophyll fluorescence


non-photochemical quenching of chlorophyll fluorescence




Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Anderson, L.E., Duggan, J.X. (1977) Inhibition of light modulation of chloroplast enzyme activity by sulfite. Oecologia 28, 147–151Google Scholar
  2. Arnon, D.I. (1949) Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiol. 24, 1–15Google Scholar
  3. Badger, M.R., Sharkey, T.D., v. Caemmerer, S. (1984) The relationship between steady state gas exchange of bean leaves and the levels of carbon-reduction-cycle intermediates. Planta 160, 305–313Google Scholar
  4. Bradbury, M., Baker, N.R. (1981) Analysis of the slow phases of the in vivo chlorophyll fluorescence induction curve. Changes in the redox state of photosystem II electron acceptors and fluorescence emission from photosystems I and II. Biochim. Biophys. Acta 635, 542–551Google Scholar
  5. Beyschlag, W., Wedler, M., Lange, O.L., Heber, U. (1987) Einfluß einer Magnesiumdüngung auf Photosynthese und Transpiration von Fichten an einem Magnesium-Mangelstandort im Fichtelgebirge. Allg. Forst. Z. 27/28/29, 738–741Google Scholar
  6. Bilger, W., Björkman, O. (1990) Role of the xanthophyll cycle in photoprotection elucidated by measurements of light-induced absorbance changes, fluorescence and photosynthesis in leaves of Hedera canariensis. Photosynth. Res. 25, 173–185Google Scholar
  7. Bilger, W., Heber, U., Schreiber, U. (1988) Kinetic relationships between energy-dependent fluorescence quenching, light scattering, chlorophyll luminescence and proton pumping in intact leaves. Z. Naturforsch. 43c, 877–887Google Scholar
  8. Bilger, W., Björkman, O., Thayer, S.S. (1989) Light-induced spectral absorbance changes in relation to photosynthesis and the epoxidation state of xanthophyll cycle components in cotton leaves. Plant Physiol. 91, 542–551Google Scholar
  9. Cerovic, Z.G., Kalezic, R., Plesnicar, M. (1982) The role of photophosphorylation in SO2 and SO32− inhibition of photosynthesis in isolated chloroplasts. Planta 156, 249–254Google Scholar
  10. Daniell, H., Sarojini, G. (1981) On the possible site of sulfite action in the photosynthetic electron transport chain and the light modulation of enzyme activity. Photobiochem. Photobiophys. 2, 61–68Google Scholar
  11. Demmig, B., Winter, K., Krüger, A., Czygan, F.-C. (1987) Photoinhibition and zeaxanthin formation in intact leaves. A possible role of the xanthophyll cycle in the dissipation of excess light energy. Plant Physiol. 84, 218–224Google Scholar
  12. Dietz, K.-J., Heber, U. (1984) Rate limiting factors in photosyn-thesis. I. Carbon fluxes in the Calvin cycle. Biochim. Biophys. Acta 767, 432–443Google Scholar
  13. Dietz, K.-J., Schreiber, U., Heber, U. (1985) The relationship between the redox state of QA and photosynthesis in leaves at various carbon-dioxide, oxygen and light regimes. Planta 166, 219–226Google Scholar
  14. Dittrich, A.P.M., Pfanz, H., Heber, U. (1992) Oxidation and reduction of SO2 by chloroplasts and formation of sulfite addition compounds. Plant Physiol 98, 738–744Google Scholar
  15. Genty, B., Briantais, J.-M., Baker, N.R. (1989) The relationship between the quantum yield of photosynthetic electron transport and photochemical quenching of chlorophyll fluorescence. Biochim. Biophys. Acta 990, 87–92Google Scholar
  16. Ghisi, R., Dittrich, A.P.M., Heber, U. (1990) Oxidation versus reductive detoxification of SO2 by chloroplasts. Plant Physiol. 92, 846–849Google Scholar
  17. Gilmore, A.M., Yamamoto, H.Y. (1991) Resolution of lutein and zeaxanthin using a non-endcapped lightly carbon-loaded C18 high-performance liquid chromatographic column. J. Chromatogr. 543, 137–145Google Scholar
  18. Hager, A. (1980) The reversible, light induced conversions of xanthophylls in the chloroplasts. In: Pigments in plants, pp. 57–79, Czygan, F.-C., ed. Fischer, StuttgartGoogle Scholar
  19. Heber, U. (1969) Conformational changes of chloroplasts induced by illumination of leaves in vivo. Biochim. Biophys. Acta 180, 302–319Google Scholar
  20. Heber, U., Willenbrink, J. (1964) Sites of synthesis and transport of photosynthetic products within the leaf cell. Biochim. Biophys. Acta 82, 313–324Google Scholar
  21. Kaiser, W.M., Dittrich, A.P.M., Heber, U. (1991) Sulfatakkumulation in Fichtennadeln als Folge von SO2-Belastung. In: Proceedings. 2. Statuseminar der PBWU zum Forschungsschwerpunkt “Waldschäden”. GSF Bericht 26/91, pp. 425–438, GSF Forschungszentrum für Umwelt und Gesundheit GmbH., NeuherbergGoogle Scholar
  22. Klughammer, C., Schreiber, U. (1991) Analysis of light-induced absorbance changes in the near-infrared spectral region. I. Characterization of various components in isolated chloroplasts. Z. Naturforsch. 46c, 233–244Google Scholar
  23. Kobayashi, V., Köster, S., Heber, U. (1982) Light scattering, chlorophyll fluorescence and state of adenylate system in illuminated spinach leaves. Biochim. Biophys. Acta 682, 44–52Google Scholar
  24. Köster, S., Heber, U. (1982) Light scattering and quenching of 9-aminoacridine fluorescence as indicators of the phosphorylation state of the adenylate system in intact chloroplasts. Biochim. Biophys. Acta 680, 88–94Google Scholar
  25. Krause, G.H. (1973) The high-energy state of the thylakoid system as indicated by chlorophyll fluorescence and chloroplast shrinkage. Biochim. Biophys. Acta 292, 715–728Google Scholar
  26. Krause, G.H., Weis, E. (1991) Chlorophyll fluorescence and photosynthesis: the basics. Annu. Rev. Plant Physiol. Plant. Mol. Biol. 42, 313–349Google Scholar
  27. Laisk, A., Oja, V., Kiirats, O., Raschke, K., Heber, U. (1989) The state of the photosynthetic apparatus in leaves as analyzed by rapid gas exchange and optical methods: the pH of the chloroplast stroma, and activation of enzymes in vivo. Planta 177, 350–358Google Scholar
  28. Nobel, P.S. (1983) Biophysical plant physiology and ecology. W.H. Freeman, New YorkGoogle Scholar
  29. Schreiber, U., Schliwa, U., Bilger, W. (1986) Continuous recording of photochemical and non-photochemical fluorescence quenching with a new type of modulation fluorometer. Photosynth. Res. 10, 51–62Google Scholar
  30. 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, 686–698Google Scholar
  31. Siebke, K., Laisk, A., Oja, V., Kiirats, O., Raschke, K., Heber, U. (1990) Control of photosynthesis in leaves as revealed by rapid gas exchange and measurements of the assimilatory force FA. Planta 182, 513–522Google Scholar
  32. Shimazaki, K., Nakamachi, K., Kondo, N., Sugahara, K. (1984) Sulfite inhibition of photosystem II in illuminated spinach leaves. Plant Cell Physiol. 25, 337–341Google Scholar
  33. Slovik, S., Kaiser, W.M., Körner, Ch., Kindermann, G., Heber, U. (1992) Quantifizierung der physiologischen Kausalkette von SO2-Immissionsschäden für Rotfichten (Picea abies (L.) Karst.). II. Ableitung von SO2-Immissionsgrenzwerten für chronische Schäden. Allg. Forst Z. 17, 913–920Google Scholar
  34. Tanaka, K., Otsubo, T., Kondo, N. (1983) Participation of hydrogen peroxide in the inactivation of Calvin cycle enzymes in SO2-fumigated leaves. Plant Cell Physiol. 23, 1009–1018Google Scholar
  35. Tanaka, K., Mitsuhashi, H., Kondo, N., Sugahara, K. (1984) Further evidence for inactivation of fructose-1,6-bisphosphatase at the begining of SO2 fumigation. Increase in fructose-1,6-bisphosphate and decrease in fructose-6-phosphate in SO2-fumigated spinach leaves. Res. Report Natl. Inst. Environ. Stud. Jpn. 65, 213–216Google Scholar
  36. Taylor, G.E. jr, Tingey, D.T. (1983) Sulfur dioxide flux into leaves of Geranium carolinianum L. Plant Physiol. 72, 237–244Google Scholar
  37. Thaler, M., Simonis, W., Schönknecht, G. (1992) Light-dependent changes of the cytoplasmic H+ and Cl- activity in the green alga Eremosphaera viridis. Plant Physiol. 99, 103–110Google Scholar
  38. Vernon, L.P. (1960) Spectrophotometric determination of chlorophylls and phaeophytins in plant extracts. Anal. Chem. 32, 1144–1150Google Scholar
  39. van Kooten, O., Snel, J.F.H. (1990) The use of chlorophyll fluorescence nomenclature in plant stress physiology. Photosynth. Res. 25, 147–150Google Scholar
  40. Wagner, U., Kolbowski, J, Oja, V., Laisk, A., Heber, U. (1990) pH homeostasis of the chloroplast stroma can protect photosynthesis of leaves during the influx of potentially acidic gases. Biochim. Biophys. Acta 1016, 115–120Google Scholar
  41. Winner, W.E., Mooney, H.A., Goldstein, R.A. (1985) Sulfur dioxide and Vegetation, Stanford University Press, StanfordGoogle Scholar
  42. Ziegler, I. (1973) The effect of SO3 on the activity of ribulose-1,5-diphosphate carboxylase in isolated spinach chloroplasts. Planta 103, 155–163Google Scholar
  43. Yin, Z.H., Neimanis, S., Wagner, U., Heber, U. (1990) Light-dependent pH changes in leaves of C3 plants. I. Recording pH changes in different cellular compartments by fluorescent probes. Planta 182, 244–252Google Scholar

Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • Sonja Veljovic-Jovanovic
    • 1
  • Wolfgang Bilger
    • 1
  • Ulrich Heber
    • 1
  1. 1.Julius-von-Sachs Institut für Biowissenschaften, Universität WürzburgWürzburgGermany

Personalised recommendations