Advertisement

Photosynthesis Research

, Volume 58, Issue 2, pp 123–133 | Cite as

In situ detection of heavy metal substituted chlorophylls in water plants

  • Hendrik Küpper
  • Frithjof Küpper
  • Martin Spiller
Article

Abstract

The in vivo substitution of magnesium, the central atom of chlorophyll, by heavy metals (mercury, copper, cadmium, nickel, zinc, lead) leads to a breakdown in photosynthesis and is an important damage mechanism in heavy metal-stressed plants. In this study, a number of methods are presented for the efficient in situ detection of this substitution (i.e. in whole plants or in chloroplasts). While macroscopic observations point to the formation of heavy metal chlorophylls at higher concentrations, fluorescence microscopy enables the detection of this reaction at very low substitution rates. Therefore, the course of the reaction can be followed by continuously measuring the fluorescence of whole plants. Furthermore absorbance spectroscopy of whole cells or isolated chloroplasts also enables the in situ detection of heavy metal chlorophylls. These methods provide practicable approaches in detecting the formation of these compounds in situ, avoiding artefacts that might occur using extraction methods based on polar solvents. In addition to the new methods for in situ detection, an extreme heterogeneity in the reaction of cells in the same tissue upon heavy metal stress was observed: while some cells are already disintegrating, others still show normal fluorescence and photosynthetic activity. Measurements of fluorescence kinetics gave a further hint that in high light intensity a substitution of Mg by heavy metals might take place specifically in PS II reaction centres.

antenna pigments copper fluorescence microscopy fluorimetry spectroscopy zinc 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Allen KD and Staehelin LA (1991) Resolution of 16 to 20 chlorophyll–protein complexes using a low ionic strength, green gel system. Anal Biochem 194: 214–222Google Scholar
  2. Atal N, Saradhi PP and Mohanty P (1991) Inhibition of the chloroplast photochemical reactions by treatment of wheat seedlings with low concentrations of cadmium: Analysis of electron transport activities and changes in fluorescence yield. Plant Cell Physiol 32(7): 943–951Google Scholar
  3. Bernier M, Popovic R and Carpentier R (1993) Mercury inhibition at the donor side of photosystem II is reversed by chloride. FEBS Lett 321(1): 19–23Google Scholar
  4. Clijsters H and Van Assche F (1985) Inhibition of photosynthesis by heavy metals. Photosynth Res 7: 31–40Google Scholar
  5. De Filippis LF (1979) The effect of heavy metals on the absorption spectra of Chlorella cells and chlorophyll solutions. Z Pflanzenphysiol 93: 129–37Google Scholar
  6. El-Sheekh MM(1993) Inhibition of Photosystem II in the green alga Scenedesmus obliquus by nickel. Biochem Physiol Pflanz 188: 363–272Google Scholar
  7. Fernandes JC and Henriques FS (1991) Biochemical, physiological and structural effects of excess copper in plants. Bot Rev 57(3): 246–273Google Scholar
  8. Furuya K (1990) Subsurface chlorophyll maximum in the tropical and subtropical western Pacific Ocean: Vertical profiles of phytoplankton biomass and its relationship with chlorophyll a and particulate organic carbon. Mar Biol 107: 529–539Google Scholar
  9. Gaudet T (1963) Marsilea vestita: Conversion of the water form to the land form by darkness and by far-red light. Science 140: 975–976Google Scholar
  10. Georgiou GN, Ahmet MT, Houlton A, Silver J and Cherry RJ (1994) Measurement of the rate of uptake and subcellular localization of porphyrins in cells using fluorescence digital imaging microscopy. Photochem Photobiol 59(4): 419–422Google Scholar
  11. Gross RE, Pugno P and Dugger WM (1970) Observations on the mechanism of copper damage in Chlorella. Plant Physiol 46, 183–185Google Scholar
  12. Hall JA (1991) Long-term preservation of picophytoplankton for counting by fluorescence microscopy. Br Phycol J 26: 169–174Google Scholar
  13. Karukstis KK (1991) Chlorophyll fluorescence as a physiological probe of the photosynthetic apparatus. In: Scheer H (ed) Chlorophylls, pp 770–797. CRC Press, Boca Raton/Ann Arbor/ Boston/LondonGoogle Scholar
  14. Kimimura M and Katoh S (1972) Studies on electron transport associated with Photosystem I: I. Functional site of plastocyanin, inhibitory effects of HgCl2 on electron transport and plastocyanin in chloroplasts. Biochim Biophys Acta 283: 279–292Google Scholar
  15. Kowalewska G and Hoffmann SK (1989) Identification of the copper porphyrin complex formed in cultures of blue-green alga Anabaena variabilis. Acta Physiol Planta 11: 39–50Google Scholar
  16. Krause G and Weis E (1984) Chlorophyll fluorescence as a tool in plant physiology. II: Interpretation of fluorescence signals. Photosynth Res 5: 139–157Google Scholar
  17. Krupa Z, Öquist G and Huner NPA (1993) The effects of cadmium on photosynthesis of Phaseolus vulgaris – fluorescence analysis. Physiol Planta 88: 626–630Google Scholar
  18. Küpper H, Küpper F and Spiller M (1996) Environmental relevance of heavy metal substituted chlorophylls using the example of water plants. J Exp Bot 47(295): 259–266Google Scholar
  19. Lanaras T, Moustakas M, Symeonidis L, Diamantoglou S and Karataglis S (1993) Plant metal content, growth responses and some photosynthetic measurements on field-cultivated wheat growing on ore bodies enriched in Cu. Physiol Planta 88: 307–314Google Scholar
  20. Lehnen LP Jr, Sherman TD, Becerril JM and Duke SO (1990) Tissue and cellular localization of acifluorfen-induced porphyrins in cucumber cotyledons. Pesticide Biochem Physiol 37: 239–248Google Scholar
  21. Lichtenthaler HK and Rinderle U (1988) Role of chlorophyll fluorescence in the detection of stress conditions in plants. CRC Crit Rev Anal Chem 19 Suppl 1: 29–85Google Scholar
  22. Li EH and Miles CD (1975) Effects of cadmium on photoreaction II of chloroplasts. Plant Sci Lett 5: 33–40Google Scholar
  23. Lidon FC, Ramalho JC and Henriques FS (1993) Copper inhibition of rice photosynthesis. J Plant Physiol 142: 12–17Google Scholar
  24. Markert B (ed) (1993) Plants as Biomonitors: Indicators for Heavy Metals in the Terrestrial Environment. VCH Verlagsgesellschaft, WeinheimGoogle Scholar
  25. Ouzounidou G (1993) Changes in variable chlorophyll fluorescence as a result of Cu-treatment: Dose-response relations in Silene and Thlaspi. Photosynthetica 29(3): 455–462Google Scholar
  26. Pirson A (1958) Die mineralische Ernährung der Pflanze. In: Adriani MJ and Baumeister W et al. (eds) Encyclopedia of Plant Physiology/Handbuch der Pflanzenphysiologie, Vol IV. Springer, BerlinGoogle Scholar
  27. Puckett KJ (1976) The effects of heavy metals on some aspects of lichen physiology. Can J Bot 54: 2695–2703Google Scholar
  28. Samuelsson G and Öquist G (1980) Effects of copper chloride on photosynthetic electron transport and chlorophyll–protein complexes of Spinacia oleracea. Plant Cell Physiol 21(3): 445–454Google Scholar
  29. Sandmann G and Böger P (1980) Copper-mediated lipid peroxidation processes in photosynthetic membranes. Plant Physiol 66: 797–800Google Scholar
  30. Sgardelis S, Cook CM, Pantis JD and Lanaras T (1994) Comparison of chlorophyll fluorescence and some heavy metal concentrations in Sonchus spp. and Taraxacum spp. along an urban pollution gradient. Sci Total Environ 158:157–164Google Scholar
  31. Sims JT (1986) Soil pH effects on the distribution and plant availability of manganese, copper, and zinc. Soil Sci Soc Am J 50: 363–373Google Scholar
  32. Tripathy BC, Bhatia B and Mohanty P (1981) Inactivation of chloroplast electron-transport by Ni2+. Biochim Biophys Acta 638: 217–224Google Scholar
  33. Tsuji T, Ohki K and Fujita Y (1986) Determination of photosynthetic pigment composition in an individual phytoplankton cell in seas and lakes using fluorescence microscopy, properties of the fluorescence emitted from picophytoplankton cells. Mar Biol 93: 343–349Google Scholar
  34. Wakao N, Yokoi N, Isoyama N, Hiraishi A, Shimada K, Kobayashi M, Kise H, Iwaki M, Itoh S, Takaichi S and Sakurai Y (1996) Discovery of natural photosynthesis using Zn-containing bacteriochlorophyll in an aerobic bacterium Acidiphilium rubrum. Plant Cell Physiol 37(6): 889–893Google Scholar
  35. Walker D (1988) The Use of the Oxygen Electrode and Fluorescence Probes in Simple Measurements of Photosynthesis. Robert Hill Institute, University of Sheffield, SheffieldGoogle Scholar
  36. Watanabe T, Machida K, Suzuki H, Kobayashi M and Honda K (1985) Photoelectrochemistry of metallochlorophylls. Coordination Chem Rev 64: 207–224Google Scholar
  37. Watanabe T and Kobayashi M (1988) Chlorophylls as functional molecules in photosynthesis. Molecular composition in vivo and physical chemistry in vitro. Special Articles on Coordination Chemistry of Biologically Important Substances 4: 383–395Google Scholar
  38. Wu JT and Lorenzen H (1984) Effect of copper on photosynthesis in synchronous Chlorella cells. Bot Bull Acad Sin 25: 125–132Google Scholar

Copyright information

© Kluwer Academic Publishers 1998

Authors and Affiliations

  • Hendrik Küpper
    • 1
  • Frithjof Küpper
    • 2
  • Martin Spiller
    • 1
  1. 1.Fakultät BiologieUniversität KonstanzKonstanzGermany
  2. 2.Station BiologiqueCNRS UPR 9042RoscoffFrance

Personalised recommendations