Skip to main content

Advertisement

SpringerLink
Log in

Cadmium chelation by frustulins: a novel metal tolerance mechanism in Nitzschia palea (Kützing) W. Smith

  • Published:
Ecotoxicology Aims and scope Submit manuscript

Abstract

The ubiquity of diatom distribution, species richness, short generation time, and specific sensitivity to several environmental stressors such as metals, make diatoms particularly useful for scientific studies. Anthropogenic activities have increased the concentration of metals in air, soil and water. Due to their toxicity and persistent character, the effects of metals on organisms have been extensively studied. In this work, the association of cadmium to different extracellular molecules of Nitzschia palea cells was investigated. Cells were grown in the absence and presence (0.2 mg l−1) of cadmium in Chu no. 10 medium. Extracellular polysaccharides were extracted, and subsamples were used for polysaccharide and Cd determination. The frustules were broken mechanically under liquid nitrogen and the intracellular and frustule fractions separated. Frustulins, a protein family found on the outmost frustule layer, constituting a protection coating to environmental stress, were extracted. In each fraction proteins were quantified by the BCA method and separated by gel electrophoresis (SDS-PAGE). Cadmium associated to each fraction was quantified by Inductively Coupled Plasma Mass Spectrometry (ICP-MS) analysis. Exposure of Nitzschia palea to cadmium decreased extracellular polysaccharides by 52.8 % and increased 6 times the amount of frustulins. Cadmium was mostly retained extracellularly: 85.4 % was bound to the frustulin fraction, and 11.1 % to polysaccharides. The ability of Nitzschia palea to increase the production of frustulins due to the presence of Cd, the extracellular location of this frustulin coating and the ability of these proteins to bind Cd, suggests a new cellular defense mechanism to metals unknown until now.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Alverson A (2007) Strong purifying selection in the silicon transporters of marine and freshwater diatoms. Limnol Oceanogr 52(4):1420–1429

    Article  CAS  Google Scholar 

  • Anderson M, Gorley R, Clarke K (2008) PERMANOVA+ for PRIMER: guide to software and statistical methods. Edited by University of Auckland and PRIMER-E, Plymouth

  • Armbrust E, Berges A, Bowler C, Green B, Martinez D et al (2004) The genome of the diatom Thalassiosira pseudonana: ecology, evolution, and metabolism. Science 306:79–86

    Article  CAS  Google Scholar 

  • Bowler C, Allen A, Badger J, Grimwood J, Jabbari K et al (2008) The Phaeodactylum genome reveals the evolutionary history of diatom genomes. Nature 456:239–244

    Article  CAS  Google Scholar 

  • Branco D, Lima A, Almeida S, Figueira E (2010) Sensitivity of biochemical markers to evaluate cadmium stress in the freshwater diatom Nitzschia palea (Kützing) W. Smith. Aquat Toxicol 99(2):109–117

    Article  CAS  Google Scholar 

  • Chu S (1942) The influence of the mineral composition of the medium on the growth of planktonic algae. 1. Methods and culture media. J Ecol 31:109–148

    Google Scholar 

  • Clarke K, Gorley R (2006) PRIMER v6: user manual/tutorial. PRIMER-E, Plymouth, p 190

    Google Scholar 

  • De Filippis L, Pallaghy C (1994) Heavy metals: sources and biological effects. In: Rai LC, Gaur JP, Soeder CJ (eds) Algae and water pollution. E. Schweizerbart’sche Verlagsbuchhandlung, Stuttgart, pp 31–77

    Google Scholar 

  • Desai S, Verlecar XN, Nagarajappa, Goswami U (2006) Genotoxicity of cadmium in marine diatom Chaetoceros tenuissimus using the alkaline Comet assay. Ecotoxicology 15:359–363

    Article  CAS  Google Scholar 

  • Dobson S (1992) EHC-Environmental Health Criteria Monographs. Copyright International Programme on Chemical Safety (IPCS). Institute of Terrestrial Ecology, United Kingdom World Health Organization Geneva

  • Dubois M, Gilles K, Hamilton J, Reabers P, Smith F (1956) Colorimetric method for the determination of sugars and related substances. Anal Chem 28:350–356

    Article  CAS  Google Scholar 

  • Falasco E, Bona F, Badino G, Hoffmann L, Ector L (2009) Diatom teratological forms and environmental alterations: a review. Hydrobiologia 623:1–35

    Article  CAS  Google Scholar 

  • Figueira E, Lima A, Pereira A (2005) Monitoring glutathione levels as a marker for cadmium stress in Rhizobium leguminosarum biovar viciae. Can J Microbiol 51:7–14

    Article  CAS  Google Scholar 

  • Fisher N (1981) On the selection for heavy metal tolerance in diatoms from the Derwent Estuary, Tasmania. Austr J Mar Freshw Res 32:555–561

    Article  CAS  Google Scholar 

  • Fisher N (1982) Bioaccumulation of technetium by marine phytoplankton. Environ Sci Technol 16:579–581

    Article  CAS  Google Scholar 

  • Fisher N, Fabris J (1982) Complexation of Cu, Zn and Cd by metabolites excreted from marine diatoms. Mar Chem 11:245–255

    Article  CAS  Google Scholar 

  • Fisher N, Frood D (1980) Heavy metals and marine diatoms: influence of dissolved organic compounds on toxicity and selection for metal tolerance among four species. Mar Biol 59:85–93

    Article  CAS  Google Scholar 

  • Fisher N, Jones G (1981) Heavy metals and marine phytoplankton: correlation of toxicity and sulfhydryl-binding. J Phycol 17:108–111

    Article  CAS  Google Scholar 

  • Fisher N, Burns K, Cherry R, Heyraud M (1983) Accumulation and cellular distribution of 241Am, 210Po, and 210Pb in two marine algae. Mar Ecol Prog Ser 11:233–237

    Article  CAS  Google Scholar 

  • Fisher N, Bohé J, Teyssie L (1984) Accumulation and toxicity of Cd, Zn, Ag, and Hg in four marine phytoplankters. Mar Ecol Prog Ser 18:201–213

    Article  CAS  Google Scholar 

  • Friberg L, Nordberg G, Vouk V (1986) Handbook of the toxicology of metals, vol II. Elsevier, Amsterdam, pp 130–184

    Google Scholar 

  • Frigeri L, Radabaugh T, Haynes P, Hildebrand M (2006) Identification of proteins from a cell wall fraction of the diatom Thalassiosira pseudonana: insights into silica structure formation. Mol Cell Proteomics 5:182–193

    CAS  Google Scholar 

  • Hames B (1981) An introduction to polyacrylamide gel electrophoresis. In: Hames BD, Rickwood D (eds) Gel electrophoresis of proteins: a practical approach. IRL Press, Washington, DC, pp 1–91

  • Hildebrand M, Wetherbee R (2003) Components and control of silicification in diatoms. In: Müller WEG (ed) Silicon biomineralization. Biology, biochemistry, molecular biology, biotechnology (Progress in molecular and subcellular biology, vol 33). Springer, New York, pp 11–58

  • Jin X, Yang X, Islam E, Liu D, Mahmooda Q (2008) Effects of cadmium on ultrastructure and antioxidative defense system in hyperaccumulator and non hyperaccumulator ecotypes of Sedum alfredii Hance. J Hazard Mater 156:387–397

    Article  CAS  Google Scholar 

  • Kröger N, Poulsen N (2008) Diatoms—from cell wall biogenesis to nanotechnology. Annu Rev Genet 42:83–107

    Article  Google Scholar 

  • Kröger N, Bergsdorf C, Sumper M (1994) A new calcium-binding glycoprotein family constitutes a major diatom cell wall component. EMBO J 13:4676–4683

    Google Scholar 

  • Kröger N, Bergsdorf C, Sumper M (1996) Frustulins: domain conservation in a protein family associated with diatom cell walls. Eur J Biochem 239:259–264

    Article  Google Scholar 

  • Kröger N, Lehmann G, Rachel R, Sumper M (1997) Characterization of a 200-kDa diatom protein that is specifically associated with a silica-based substructure of the cell wall. Eur J Biochem 250:99–105

    Article  Google Scholar 

  • Laemmli U (1970) Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature 227:680–685

    Article  CAS  Google Scholar 

  • Lane T, Saito M, George G, Pickering I, Prince R, Morel F (2005) Isolation and Preliminary characterization of a cadmium carbonic anhydrase from a marine diatom. Nature 435:42–142

    Article  CAS  Google Scholar 

  • Lehninger A (1970) Biochemistry. Worth, New York, p 833

    Google Scholar 

  • Malik A (2004) Metal bioremediation through growing cells. Environ Int 30:261–278

    Article  CAS  Google Scholar 

  • Morelli E, Scarano G (2004) Copper-induced changes of non-protein thiols and antioxidant enzymes in the marine microalga Phaeodactylum tricornutum. Plant Sci 167:289–296

    Article  CAS  Google Scholar 

  • Morin S (2003) Amélioration des techniques de bioindication diatomique et d’analyse des données, appliquées à la révélation des effets des pollutions à toxiques. ENITA de Bordeaux: 70 pp. + annexes

  • Pistocchi R, Guerrini F, Balboni V, Boni L (1997) Copper toxicity and carbohydrate production in the microalgae Cylindrotheca fusiformis and Gymnodinium sp. Eur J Phycol 32:125–132

    Google Scholar 

  • Pistocchi R, Mormile A, Guerrini F, Isani G, Boni L (2000) Increased production of extra- and intracellular metal-ligands in phytoplankton exposed to copper and cadmium. J Appl Phycol 12:469–477

    Article  CAS  Google Scholar 

  • Price N, Morel F (1990) Cadmium and cobalt substitution for zinc in a marine diatom. Nature 344:658–660

    Article  CAS  Google Scholar 

  • Qiu R, Zhao X, Tang Y, Yu F, Hu P (2008) Antioxidative response to Cd in a newly discovered cadmium hyperaccumulator, Arabis paniculata F. Chemosphere 74:6–12

    Article  CAS  Google Scholar 

  • Staats N, De Winder Ben, Stal L, Mur L (1999) Isolation and characterization of extracellular polysaccharides from the epipelic diatoms Cylindrotheca closterium and Navicula salinarum. Eur J Phycol 34(2):161–169

    Article  Google Scholar 

  • Sunda W (1989) Trace metal interactions with marine phytoplankton. Biol Oceanogr 6:411–442

    Google Scholar 

  • Torres E, Cid A, Herrero C, Abalde J (2000) Effect of cadmium on growth, ATP content, carbon fixation and ultrastructure in the marine diatom Phaeodactylum tricornutum Bohlin. Water Air Soil Pollut 117:1–14

    Article  CAS  Google Scholar 

  • Tripathi B, Mehta S, Amar A, Gaur J (2006) Oxidative stress in Scenedesmus sp. during short- and long-term exposure to Cu2+ and Zn2+. Chemosphere 62:538–544

    Article  CAS  Google Scholar 

  • Van de Poll W, Vrieling E, Gieskes W (1999) Location and expression of frustulins in the pennate diatoms Cylindrotheca fusiformis, Navicula pelliculosa, and Navicula salinarum (Bacillariophyceae). J Phycol 35:1044–1053

    Article  Google Scholar 

  • Wenzl S, Hett R, Richthammer P, Sumper M (2008) Silacidins: highly acidic phosphopeptides from diatom shells assist in silica precipitation in vitro. Angew Chem Int Ed 120:1729–1732

    Article  Google Scholar 

  • Wong Po Foo C, Huang J, Kaplan D (2004) Lessons from seashells: silica mineralization via protein templating. Trends Biotechnol 22:577–585

    Article  Google Scholar 

Download references

Acknowledgments

We would like to thank Ana Luís for collecting N. palea used in the present study, Diana Branco for helping amongst other tasks, in the growth, harvest and counting of diatom cells, and to Ana Lima for helping with the optimization of the SDS-PAGE gels.

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. Biology Department, and CBC (Centro de Biologia Celular), University of Aveiro, Campus Universitário de Santiago, 3810-193, Aveiro, Portugal

    José Santos & Etelvina Figueira

  2. Biology Department, and GeoBioTec (GeoBioSciences, GeoTechnologies and GeoEngineering Research Center), University of Aveiro, Campus Universitário de Santiago, 3810-193, Aveiro, Portugal

    Salomé F. P. Almeida

Authors
  1. José Santos
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Salomé F. P. Almeida
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Etelvina Figueira
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Etelvina Figueira.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Santos, J., Almeida, S.F.P. & Figueira, E. Cadmium chelation by frustulins: a novel metal tolerance mechanism in Nitzschia palea (Kützing) W. Smith. Ecotoxicology 22, 166–173 (2013). https://doi.org/10.1007/s10646-012-1013-1

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10646-012-1013-1

Keywords

Search

Navigation