Skip to main content
Log in

Thiol pools and glutathione redox ratios as possible indicators of copper toxicity in the green macroalgae Enteromorpha spp. from the Scheldt Estuary (SW Netherlands, Belgium) and Thermaikos Gulf (Greece, N Aegean Sea)

  • Published:
Hydrobiologia Aims and scope Submit manuscript

Abstract

Defence mechanisms against Cu toxicity were examined in two dominant Enteromorpha species from two coastal water types. The macroalgae were collected at three locations in the eulittoral of the Scheldt Estuary (Netherlands, Belgium) and the Thermaikos Gulf (Greece). For 10 days E. prolifera (Scheldt) and E. linza (Thermaikos) were incubated in seawater media of different salinities: 6, 9, 23 psu and 25, 30, 35 psu, respectively. In one series, media were enriched with 100 μg Cu l-1; responses were compared with those in controls with no extra Cu added. Enteromorpha, which is frequently used as a monitor species for heavy metal contamination, had relatively high Cu tissue levels (0.5–3.8 μmol Cu gdwt-1). Cu levels in E. prolifera controls (Scheldt) decreased with salinity; this was not the case with Cu levels in E. linza controls (Thermaikos). During the 10-d incubation algal protein contents and tissue Cu were rather stable. In E. linza (Thermaikos) algal protein contents were significantly lower than those of E. prolifera (Scheldt), although there was no indication for nitrogen limitation in E. linza. E. linza also had much lower glutathione pools than E. prolifera. Only under acute Cu stress (metal addition) did E. prolifera synthesise metal-binding thiols (phytochelatins). Phytochelatin pools are not suitable as an indicator of the Cu levels in these algae. The glutathione redox ratio GSH:(GSH + 0.5GSSG) was used as an indicator of (Cu-induced) oxidative stress. In E. prolifera (Scheldt) this ratio decreased with algal Cu content (P <0.05), from ~0.5 to ~0.2. The average glutathione ratios in Enteromorpha from the Scheldt and Thermaikos showed some oxidative stress induction with increasing algal Cu contents, however more clearly if Cu was added. As this redox ratio can also be influenced by environmental factors such as irradiance and desiccation, it may not be useful as an indicator for Cu-induced oxidative stress in situ.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Agrawal, S. B., 1992. Effects of supplemental U.V.-B radiation on photosynthetic pigment, protein and glutathione contents in green algae. Envir. Exp. Bot. 32: 137–143.

    Article  CAS  Google Scholar 

  • Anderson, M. E., 1985. Tissue glutathione. In Greenwald, R. A. (ed.), Handbook of Methods for Oxygen Radical Research. CRC Press, Boca Raton: 317–323.

    Google Scholar 

  • Brown, L. N., M. G. Robinson & B. D. Hall, 1988. Mechanisms for copper tolerance in Amphora coffaeformis - internal and external binding. Mar. Biol. 97: 581–586.

    Article  CAS  Google Scholar 

  • Clayton, J. R., Q. Dortch, S. S. Thoresen & S. I. Ahmed, 1988. Evaluation of methods for the separation and analysis of proteins and free amino acids in phytoplankton samples. J. Plankton Res. 10: 341–359.

    CAS  Google Scholar 

  • Corzo, A. & F. X. Niell, 1991. C/N ratio in response to nitrogen supply and light quality in Ulva rigida C. Agardh (Chlorophyta: Ulvophyceaea). Sci. Mar. 55: 405–411.

    Google Scholar 

  • De Knecht, J. A., P. L. M. Koevoets, J. A. C. Verkleij & W.H.O. Ernst, 1992. Evidence against a role for phytochelatins in naturally selected increased cadmium tolerance in Silene vulgaris (Moench) Garcke. New Phytol. 122: 681–688.

    Article  CAS  Google Scholar 

  • De Knecht, J. A., N. Van Baren, W. M. Ten Bookum, H. W. N. F. Sang, P. L. M. Koevoets, H. Schat & J.A.C. Verkleij, 1995. Synthesis and degradation of phytochelatins in cadmium-sensitive and cadmium-tolerant Silene vulgaris. Plant Sci. 106: 9–18.

    Article  CAS  Google Scholar 

  • De Vos, C. H. R., M. J. Vonk, R. Vooijs & H. Schat, 1992. Glutathione depletion due to copper induced phytochelatin synthesis causes oxidative stress in Silene cucubalis. Plant Physiol. 98: 853–858.

    PubMed  CAS  Google Scholar 

  • Gekeler, W., E. Grill, E. Winnacker & M. H. Zenk, 1988. Algae sequester heavy metals via synthesis of phytochelatin complexes. Arch. Mikrobiol. 150: 197–202.

    CAS  Google Scholar 

  • Gledhill, M., M. Nimmo, S. J. Hill & M. Brown, 1997. The toxicity of copper (II) species to marine algae, with particular reference to macroalgae. J. Phycol. 33: 2–11.

    Article  CAS  Google Scholar 

  • Haritonidis, S., 1978. A survey of the marine algae of Thermaikos Gulf, Thessaloniki, Greece. I. Distribution and seasonal periodicity. Bot. mar. XXI: 527–535.

    Article  Google Scholar 

  • Haritonidis, S. & P. Malea, 1995. Seasonal and local variation of Cr, Ni and Co concentrations in Ulva rigida C. Agardh and Enteromorpha linza (Linneaus) from Thermaikos Gulf, Greece. Envir. Pollut. 89: 319–327.

    Article  CAS  Google Scholar 

  • Haritonidis, S., 1996. Chapter 17: Greece. In Schramm, W. & P.H. Nienhuis (eds), Marine Benthic Vegetation: Recent Changes and the Effects of Eutrophication, Ecological Studies 123. Springer-Verlag, Berlin, Heidelberg: 403–419.

    Google Scholar 

  • Ho, Y. B., 1990. Ulva lactuca as bioindicator of metal contamination in intertidal waters in Hong Kong. Hydrobiologia 203: 73–81.

    Article  CAS  Google Scholar 

  • Koeman, R. P. T., 1985. The taxonomy of Ulva Linnaeus, 1753, and Enteromorpha, Link, 1820 (Chlorophyceae) in the Netherlands. PhD Thesis, University Groningen, Netherlands, Van Tienderen Press: 1–201.

    Google Scholar 

  • Miersch, J., F. Bärlocher, I. Bruns & G. J. Kraus, 1997. Effects of cadmium, copper, and zinc on growth and thiol content of aquatic hyphomycetes. Hydrobiologia 346: 77–84.

    Article  CAS  Google Scholar 

  • Müller M., M. Schirmer & J. Kettler, 1993. Use of Enteromorpha intestinalis (Chlorophyceae) for active biomonitoring of heavy metals in the Weser Estuary. Neth. J. Aquat. Ecol. 27: 189–195.

    Article  Google Scholar 

  • Nassiri, Y., J. L. Mansot, J. Wery, T. Ginsburger-Vogel & J. C. Amiard, 1997. Ultrastructural and electron energy loss spectroscopy studies of sequestration mechanisms of Cd and Cu in the marine diatom Skeletonema costatum. Arch. envir. Contam. Toxicol. 33: 147–155.

    Article  CAS  Google Scholar 

  • Nieuwenhuize, J., Y. Maas & J. J. Middelburg, 1994. Rapid analysis of organic carbon and nitrogen in particulate materials. Mar. Chem. 45: 217–224.

    Article  CAS  Google Scholar 

  • Rajendran, K., P. Sampathkumar, C. Govindasamy, M. Ganesan, R. Kannan & L. Kannan, 1993. Levels of trace metals (Mn, Fe, Cu and Zn) in some Indian seaweeds. Mar. Poll. Bull. 26: 283–285.

    Article  CAS  Google Scholar 

  • Reed R. H. & L. Moffat, 1983. Copper toxicity and copper tolerance in Enteromorpha compressa (L.) Grev. J. exp. mar. Biol. Ecol. 69: 85–103.

    Article  CAS  Google Scholar 

  • Rijstenbil, J. W. & T. C. W. Poortvliet, 1992. Copper and zinc in estuarine water: chemical speciation and bioavailability to the marine planktonic diatom Ditylum brightwellii. Envir. Toxicol. Chem. 11: 1615–1625.

    CAS  Google Scholar 

  • Rijstenbil, J. W, S. Haritonidis, P. Malea, J. van Drie & J. A. Wijnholds, 1993. Interactions of copper with trace metals and thiols in the macro-algae Enteromorpha prolifera (O.F. Müll) J. Ag., grown in water of the Scheldt Estuary (Belgium and S.W. Netherlands). Sci. Tot. Envir. Suppl. 1993: 539–549.

    Google Scholar 

  • Rijstenbil, J. W., J. W. M. Derksen, L. J. A. Gerringa, T. C. W. Poortvliet, A. Sandee, M. van den Berg, J. van Drie & J. A. Wijnholds, 1994. Oxidative stress induced by copper: defense and damage in the marine planktonic diatom Ditylum brightwellii (Grunow) West, grown in continuous cultures with high and low zinc levels. Mar. Biol. 119: 583–590.

    Article  CAS  Google Scholar 

  • Rijstenbil, J. W. & J. A. Wijnholds, 1996. HPLC analysis of nonprotein thiols in planktonic diatoms: pool size, redox state and response to copper and cadmium exposure. Mar. Biol. 127: 45- 54.

    Article  CAS  Google Scholar 

  • Rijstenbil, J. W., F. Dehairs, R. Ehrlich & J. A. Wijnholds, 1998. Effect of the nitrogen status on copper accumulation and pools of metal-binding peptides in the planktonic diatom Thalassiosira pseudonana. Aquat. Toxicol. 42: 187–209.

    Article  CAS  Google Scholar 

  • Robinson, N. J., 1989. Algal metallothioneins: secondary metabolites and proteins. J. appl. Phycol. 1: 5–18.

    Article  CAS  Google Scholar 

  • Say, P. J., I. G. Burrows & B. A. Whitton, 1990. Enteromorpha as a monitor of heavy metals in estuaries. Hydrobiologia 195: 119–126.

    Article  CAS  Google Scholar 

  • Seeliger, U. & C. Cordazzo, 1982. Field and experimental evaluation of Enteromorpha sp. as a quali-quantitative monitoring organism for copper and mercury in estuaries. Envir. Pollut. A 29: 197–206.

    Article  CAS  Google Scholar 

  • Siller-Cepeda, J. H., T. H. H. Chen & L. H. Fuchigami, 1991. High performance liquid chromatography analysis of reduced and oxidized glutathione in woody plant tissues. Plant Cell Physiol. 32: 1179–1185.

    CAS  Google Scholar 

  • Smith, I. K., A. C. Kendall, A. J. Keys, J. C. Turner & P. J. Lea, 1985. The regulation of the biosynthesis of glutathione in leaves of barley (Hordeum vulgare L.). Plant Sci. 41: 11–17.

    Article  CAS  Google Scholar 

  • Stauber, J. L. & T. M. Florence, 1987. Mechanisms of toxicity of ionic copper and copper complexes to algae. Mar. Biol. 94: 511- 519.

    Article  CAS  Google Scholar 

  • Stegenga, H. & I. Mol, 1983. Flora van de Nederlandse Zeewieren. KNNV Series 33. Erla Amsterdam Print: 1–263.

  • Steffens, J. C., D. F. Hunt & B. G. Williams, 1986. Accumulation of non-protein metal-binding polypeptides (gamma-glutamylcysteinyl) n-glycine in selected cadmium-resistent tomato cells. J. Biol. Chem. 261: 13879–13982.

    PubMed  CAS  Google Scholar 

  • Steffens, J. C., 1990. Heavy metal stress and the phytochelatin response. In Alscher, R.G. & J.R. Cumming (eds), Stress Responses in Plants: Adaptation and Acclimation Mechanisms, Vol. 12. Wiley & Liss Inc., New York, 17: 377–394.

    Google Scholar 

  • Strickland, J. D. H. & T. R. Parsons, 1972. A practical handbook of seawater analysis. Fish. Res. Bd Can. Bull. 167: 1–311.

    Google Scholar 

  • Sueur, S., C. M. G. van den Berg & J. P. Riley, 1982. Measurement of the metal complexing ability of exudates of marine macroalgae. Limnol. Oceanogr. 27: 536–543.

    Article  CAS  Google Scholar 

  • Vögeli-Lange, R. & G. J. Wagner, 1990. Subcellular localization of cadmium and cadmium-binding peptides in tobacco leaves. Implication of a transport function for cadmium-binding peptides. Plant Physiol. 92: 1086–1093.

    Article  PubMed  Google Scholar 

  • Zolotukhina, E. Y. & E. E. Gavrilenko, 1990. Binding of copper, cadmium, zinc and manganese in proteins of aquatic macrophytes. Fyziologiya Rastenii 37: 651–658.

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rijstenbil, J., Haritonidis, S., Malea, P. et al. Thiol pools and glutathione redox ratios as possible indicators of copper toxicity in the green macroalgae Enteromorpha spp. from the Scheldt Estuary (SW Netherlands, Belgium) and Thermaikos Gulf (Greece, N Aegean Sea). Hydrobiologia 385, 171–181 (1998). https://doi.org/10.1023/A:1003502428466

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1003502428466

Navigation