, Volume 192, Issue 2, pp 261–268 | Cite as

An improved method, using saturating light pulses, for the determination of photosystem I quantum yield via P700+-absorbance changes at 830 nm

  • Christof Klughammer
  • Ulrich Schreiber


An improved method is introduced for the determination of the quantum yield of photosystem I. The new method employs saturating light pulses with steep rise characteristics to distinguish, in a given physiological state, centers with an open acceptor side from centers with a reduced acceptor side. The latter do not contribute to PSI quantum yield (ΦI). Oxidation of P700 is measured by a rapid modulation technique using the absorbance change around 830 nm. The quantum yield ΦI is calculated from the amplitude of the rapid phase of absorbance change (ΔA; 830 nm) upon application of a saturation pulse in a given state, divided by the maximal ΔA (830 nm) which is induced by a saturation pulse with far-red background illumination. Using this technique, ΦI can be determined even under conditions of acceptor-side limitation, as for example in the course of a dark-light induction period or after elimination of CO2 from the gas stream. Thus determined ΦI values display a close-to-linear relationship with those for the quantum yield of PSII (ΦII) calculated from chlorophyll fluorescence parameters. It is concluded that the proposed method may provide new information on the activity of the PSI acceptor side and thus help to separate the effects of acceptorside limitation from those of cyclic PSI, whenever a non-linear relationship between ΦII and the P700-reduction level is observed.

Key words

Chlorophyll fluorescence Light absorbance (830 nm) P700 Photosynthesis Quantum yield 

Abbreviations and Symbols


absorbance change


quantum yield of photosystem I


quantum yield of photosystem II


photosynthetically active radiation


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Asada, K., Heber, U., Schreiber, U. (1993) Electron flow to intersystem chain from stromal components and cyclic electron flow in maize chloroplasts as detected in intact leaves by monitoring P700 and chlorophyll fluorescence. Plant Cell Physiol.34, 39–50Google Scholar
  2. Foyer, C., Furbank, R., Harbinson, J., Horton, P. (1990) The mechanisms contributing to photosynthetic control of electron transport by carbon assimilation in leaves. Photosynth. Res.25, 83–100Google Scholar
  3. Foyer, C.H., Lelandais, M., Harbinson, J. (1992) Control of the quantum efficiencies of photosystem I and II, electron flow, and enzyme activation following dark-to-light transitions in pea leaves. Plant Physiol.99, 979–986Google Scholar
  4. Genty, B., Briantais, J.-M., Baker, N.R. (1989) The relationship between the quantum yield of photosynthetic electron transport and quenching of chlorophyll fluorescence. Biochim. Biophys. Acta990, 87–92Google Scholar
  5. Genty, B., Wonders, J. Baker N.R. (1990) Non-photochemical quenching of Fo in leaves is emission wavelength dependent: consequences for quenching analysis and its interpretation. Photosynth. Res.26, 133–139Google Scholar
  6. Harbinson, J., Foyer, C. (1991) Relationships between the efficiencies of photosystems I and II and stromal redox state in CO2-free air. Evidence for cyclic electron flow in vivo. Plant Physiol.97, 41–49Google Scholar
  7. Harbinson, J., Hedley, C.L. (1989) The kinetics of P700 reduction in leaves: a novel in situ probe of thylakoid functioning. Plant Cell Environ.12, 357–369Google Scholar
  8. Harbinson, J., Woodward, F.I. (1987) The use of light-induced absorbance changes at 830 nm to monitor the oxidation state of P700 in leaves. Plant Cell Environ.10, 131–140Google Scholar
  9. Harbinson, J., Genty, B., Baker, N.R. (1989) Relationship between the quantum efficiencies of photosystems I and II in pea leaves. Plant Physiol.90, 1029–1034Google Scholar
  10. Harbinson, J., Genty, B., Foyer, C.H. (1990) Relationship between photosynthetic electron transport and stromal enzyme activitiy in pea leaves. Toward an understanding of the nature of photosynthetic control. Plant Physiol.94, 545–553Google Scholar
  11. Heber, U., Walker, D. (1992) Concerning a dual function of coupled cyclic electron transport in leaves. Plant Physiol.100, 1621–1626Google Scholar
  12. Heber, U., Schreiber, U., Siebke, K., Dietz, K.-J. (1990) Relationship between light-driven electron transport, carbon reduction and carbon oxidation in photosynthesis. In: Perspectives in biochemical and genetic regulation of photosynthesis, pp. 17–37, Zelitch, J., ed. Wiley-Liss, New YorkGoogle Scholar
  13. Herbert, S.K., Fork, D.C., Malkin, S.H. (1990) Photoacoustic measurements in vivo of energy storage by cyclic electron flow in algae and higher plants. Plant Physiol.94, 926–934Google Scholar
  14. Horton, P. (1989) Interactions between electron transport and carbon assimilation: regulation of light harvesting and photochemistry. In: Photosynthesis, pp. 393–406, Briggs, W.R., ed. Alan R. Liss Inc., New YorkGoogle Scholar
  15. 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
  16. Laisk, A., Oja, V., Heber, U. (1993) Steady-state and induction kinetics of photosynthetic electron transport related to donor side oxidation and acceptor side reduction of photosystem I in sunflower leaves. Photosynthetica, in pressGoogle Scholar
  17. Lechtenberg, D., Voss, B., Weis, E. (1989) Regulation of photosynthesis: Photosynthetic control and thioredoxin-dependent enzyme regulation. In: Current research in photosynthesis, vol. IV, pp. 171–174, Baltscheffsky, M., ed. Kluwer Academic Publishers, Dordrecht, The NetherlandsGoogle Scholar
  18. 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 cyanobacteriumSynechocystis PCC 6803. Plant Cell Physiol.33, 1233–1237Google Scholar
  19. Neubauer, C., Schreiber, U. (1987) The polyphasic rise of chlorophyll fluorescence upon onset of strong continuous illumination: I. Saturation characteristics and partial control by the photosystem II acceptor side. Z. Naturforsch.42c, 1246–1254Google Scholar
  20. Peterson, R.B. (1991) Effects of O2 and CO2 concentrations on quantum yields of photosystem I and II in tobacco leaf tissue. Plant Physiol.97, 1388–1394Google Scholar
  21. Scheibe, R., Stitt, M. (1988) Comparison of NADP-malate dehydrogenase activation, QA reduction and O2 evolution in spinach leaves. Plant Physiol. Biochem.26, 473–482Google Scholar
  22. Schreiber, U. (1986) Detection of rapid induction kinetics with a new type of high-frequency modulated chlorophyll fluorometer. Photosynth. Res.9, 261–272Google Scholar
  23. Schreiber, U., Schliwa, U., Bilger, W. (1986) Continuous recording of photochemical and non-photochemical chlorophyll fluorescence quenching with a new type of modulated fluorometer. Photosynth. Res.10, 51–62Google Scholar
  24. 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
  25. Van Kooten, O., Snel, J.F.H. (1990) The use of chlorophyll fluorescence nomenclature in plant stress physiology. Photosynth. Res.25, 147–150Google Scholar
  26. Walker, D.A. (1992) Excited leaves. New Phytol121, 325–345Google Scholar
  27. Weis, E., Berry, J. (1987) Quantum efficiency of photosystem II in relation to energy-dependent quenching of chlorophyll fluorescence. Biochim. Biophys. Acta894, 198–208Google Scholar
  28. Weis, E., Ball, J.R., Berry, J. (1987) Photosynthetic control of electron transport in leaves ofPhaseolus vulgaris. Evidence for regulation of photosystem 2 by the proton gradient. In: Progress in photosynthesis research, vol. 2, pp. 553–556, Biggins, J., ed. Martinus-Nijhoff Publishers, DordrechtGoogle Scholar
  29. Weis, E., Lechtenberg, D. (1989) Fluorescence analysis during steady-state photosynthesis. Philos. Trans. R. Soc. Lond. B Biol. Sci.323, 253–268Google Scholar

Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • Christof Klughammer
    • 1
  • Ulrich Schreiber
    • 1
  1. 1.Lehrstuhl Botanik I, Universität WürzburgWürzburgGermany

Personalised recommendations