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Chlorophyll fluorescence at 77 K in intact leaves: Characterization of a technique to eliminate artifacts related to self-absorption

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Abstract

Due to the optical density of photosynthetic tissues the spectral characteristics of fluorescence emitted at 77 K directly from frozen plant material are distorted by differential re-absorption of the emitted light: the emission band related to PSII can be lowered by more than 80%, relative to the PSI band and the profile of the excitation spectra becomes flattened. It is demonstrated that such distortion cannot be neglected as its extent varies from sample to sample. A technique is introduced to eliminate sample artifacts related to self-absorption: subcellular small particles are prepared from rapidly cooled leaves and then ‘diluted’ without re-thawing at a concentration corresponding to about 5 μg chlorophyll·cm−3 into a matrix consisting of ice and quartz particles. The photochemical pigment apparatus is expected to remain fixed in the in vivo state. Different kinds of plant material is used and it is demonstrated how this preparative approach allows to study the in vivo distribution of energy between the two photosystems from pure 77 K spectrofluorimetry, even when the optical properties of whole leaves or thalli normally would exclude quantitative analysis.

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References

  1. Arnon DJ (1949) Copper enzymes in isolated chloroplasts. Plant Physiol 24:1–15

    Google Scholar 

  2. Baker NR and Strasser RJ (1982) Development and spectral characteristics of excitation energy transfer between the photosystems during chloroplast biogenesis. Photobiochem Photobiophys 4:265–273

    Google Scholar 

  3. Butler WL (1964) Absorption spectroscopy in vivo: theory and application. Ann Rev Plant Physiol 15:451–470

    Google Scholar 

  4. Butler WL (1977) Chlorophyll fluorescence: A probe for electron transfer. In Trebst A and Avron M eds, Encyclopedia of Plant Physiology, New Serie Vol 5, pp 149–167, Heidelberg: Springer

    Google Scholar 

  5. Butler WL and Strasser RJ (1977) Does the rate of cooling affect fluorescence properties of chloroplasts at −196°C? Biochim Biophys Acta 462:283–289

    Google Scholar 

  6. French CS (1971) The distribution and action in photosynthesis of several forms of chlorophyll. Proc Natl Acad Sci U.S.A. 68:2893–2897

    Google Scholar 

  7. Goedheer JC (1969) Energy transfer from carotenoids to chlorophyll in blue-green, red and green algae and greening bean leaves. Biochim Biophys Acata 172:152–265

    Google Scholar 

  8. Govindjee and Yang L (1966) Structure of the red fluorescence band in chloroplasts. J Gen Physiol 49:763–780

    Google Scholar 

  9. Harnischfeger G (1977) The use of fluorescence emission at 77°K in the analysis of the photosynthetic apparatus of higher plants and algae. Advan Bot Res 5:1–52

    Google Scholar 

  10. Hodges M and Barber J (1983) Photosynthesis adaptation of pea plants grown at different light intensities: state 1-state 2 transition and associated chlorophyll fluorescence changes. Planta 157:166–175

    Google Scholar 

  11. Krause GH, Briantais J-M and Vernotte C (1983) Characterization of chlorophyll fluorescence quenching in the chloroplasts by fluorescence spectroscopy at 77 K I. ΔpH-dependent quenching. Biochim Biophys Acta 723:169–175

    Google Scholar 

  12. Krause GH and Weis E (1984) Chlorophyll fluorescence as a tool in plant physiology II. Interpretation of fluorescence signals. Photosynth Res (in press)

  13. Kitajiama M and Butler WL (1975) Excitation spectra for photosystem I and photosystem II in chloroplasts and the spectral characteristics of the distribution of quanta between the two photosystems. Biochim Biophys Acta 408:297–305

    Google Scholar 

  14. Lichtenthaler HK, Buschmann C, Döll M, Fietz HJ, Bach T, Kozel U, Meier D and Rahmsdorf U (1971) Photosynthetic activity, chloroplast ultrastructure and leaf characteristics of high-light and low-light plants and of sun and shade leaves. Photosynth Res 2:115–141

    Google Scholar 

  15. Murata N, Nishimura M and Takamiya A (1966) Fluorescence of chlorophyll in photosynthetic systems. III. Emission and action spectra of fluorescence — three emission bands of chlorophyll a and the energy transfer between the two pigment systems. Biochim Biophys Acta 126:234–243

    Google Scholar 

  16. Öquist G and Forc DC (1982) Effects of desiccation on the excitation energy distribution from phycoerytrin to the two photosystems in the red alga Porphyra perforata. Physiol Plant 56:56–62

    Google Scholar 

  17. Öquist G and Fork DC (1982) Effects of desiccation on the 77°K fluorescence properties of the liverwort Porella navicularis and the isolated lichen green alga Trebouxia pyriformis. Physiol Plant 56:63–68

    Google Scholar 

  18. Powels SB and Björkman O (1982) Photoinhibition of photosynthesis: effect on chlorophyll fluorescence at 77K in intact leaves and in chloroplasts of Nerium oleander. Planta 165:79–107

    Google Scholar 

  19. Satoh and Fork DC (1983) Changes in the distribution of light energy between the two photosystems in spinach leaves. Photosyn Res 4:71–79

    Google Scholar 

  20. Satoh K and Fork DC (1983) A new mechanism for adaptation to changes in light intensity and quality in the red alga, Porphyra perforata I. Relation to state 1-state 2 transitions. Biochim Biophys Acta 722:190–196

    Google Scholar 

  21. Strasser RJ and Butler WL (1977) Energy transfer and the distribution of excitation energy in the photosynthetic apparatus of spinach chloroplasts. Biochim Biophys Acta 460:230–238

    Google Scholar 

  22. Strasser RJ and Butler WL (1977) The yield of energy transfer and spectral distribution of excitation energy in the photochemical apparatus in flashed bean leaves. Biochim Biophys Acta 462:295–306

    Google Scholar 

  23. Szalay L, Török M and Govindjee (1967) Effect of secondary fluorescence in the emission spectrum and quantum yield of fluorescence in chlorophyll a solution and algal suspensions. Acta Biochem Biophys Acad Sci Hung 2:425–432

    Google Scholar 

  24. Weis E (1984) Short term acclimation of spinach to high temperatures. Effect on chlorophyll fluorescence at 293 and 77 Kelvin in intact leaves. Plant Physiol 74:402–407

    Google Scholar 

  25. Weis E (1984) Temperature-induced changes in the distribution of excitation energy between photosystem I and photosystem II in spinach leaves. In Sybesma C (ed.) Advances in Photosynthesis Research, Vol. 3, pp 291–294, The Hague: M. Nijhoff/Dr W. Junk Publishers

    Google Scholar 

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  1. Botanisches Institut, Universität Düsseldorf, D-4000, Düsseldorf, Fed. Rep. Germany

    Engelbert Weis

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  1. Engelbert Weis
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Weis, E. Chlorophyll fluorescence at 77 K in intact leaves: Characterization of a technique to eliminate artifacts related to self-absorption. Photosynth Res 6, 73–86 (1985). https://doi.org/10.1007/BF00029047

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  • DOI: https://doi.org/10.1007/BF00029047

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