Skip to main content

Advertisement

Log in

Phyto/rhizoremediation studies using long-term PCB-contaminated soil

  • COST ACTION 859 • PHYTOREMEDIATION • RESEARCH ARTICLE
  • Published:
Environmental Science and Pollution Research Aims and scope Submit manuscript

Abstract

Purpose

Polychlorinated biphenyls (PCBs) represent a large group of recalcitrant environmental pollutants, differing in the number of chlorine atoms bound to biphenyl ring. Due to their excellent technological properties, PCBs were used as heat-transfer media, for filling transformers and condensers, as paint additives, etc. With increasing knowledge of their toxicity, transfer to food chains and accumulation in living organisms, their production ended in most countries in the 1970s and in 1984 in the former Czechoslovakia. But even a quarter of century after the PCB production ceased, from contaminated areas, the volatile PCBs evaporate and contaminate much larger areas even at very distant parts of the world. For this reason, PCBs still represent a global problem. The main method of PCB removal from contaminated environment is at present the expensive incineration at high temperatures. With the aim of finding effective alternative approaches, we are studying biological methods for PCB removal from the environment. In this paper, we summarise 10 years of studies using long-term PCB-contaminated soil from a dumpsite in South Bohemia, targeted for the use of plants (phytoremediation) and their cooperation with microorganisms in the root zone (rhizoremediation).

Materials and methods

Long-term contaminated soil from Lhenice dumpsite, more than hundred kilograms of homogenised material, was used in microcosms (pots and buckets), and field plots were established at the site. Tested plants include among others tobacco, black nightshade, horseradish, alfalfa and willow. Aseptic plant cell and tissue cultures were from the collection of the IOCB. Microorganisms were our own isolates. The paper summarises experiments done between 1998 and 2008 with real contaminated soil, both vegetated and non-vegetated. PCB analysis was performed by GC-ECD, metabolic products identified mostly using 2D-GC/MS-MS and synthetic standards, whereas molecular methods included quantitative PCR and sequencing.

Results

The soil was used both for preparation of field plots at the site and for greenhouse and laboratory tests in microcosms. The results include analyses of changes in PCB content in untreated and vegetated soil, PCB uptake and distribution in different parts of various plant species, analysis of products formed, identification and characterisation of cultivable and non-cultivable bacteria both in rhizosphere and in bulk soil. Different treatments and amendments were also tested. Experiments in real contaminated soil were accompanied by in vitro experiments using aseptic cultures of plant biomass, genetically modified (GM) plants and bacteria, to allow identification of players responsible for PCB metabolisation in soil. The time-span of the experiments allows extrapolating some of the results and drawing conclusions concerning the effectivity of exploitation of various plant species and treatments to remove PCBs from soils.

Discussion

The approach using plants proved to represent a viable alternative to costly incineration of PCB-contaminated soils. The recent studies using molecular methods show that plants are responsible for the composition of consortia of microorganisms present in their root zone, including those with ability to degrade the chlorinated aromatic compounds.

Conclusions

In addition to uptake, accumulation and partial metabolisation of PCBs by plants, compounds produced by plants allow survival of microorganisms even in poor soils, serve as carbon and energy source, and can even induce the degradation pathways of different xenobiotics. Thus, the choice of proper plant species is crucial for effective cleaning of different polluted sites. Our study shows how the efficiency of PCB removal is dependent on the plant used.

Recommendations and perspectives

The use of plants in biological remediation of different organic xenobiotics proved to be a useful approach. Further improvement can be expected by application of specifically tailored GM plants and use of selective conditions ensuring high remediation potential based on optimal composition of the soil microbial consortia designed for the needs of given site.

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

  • Anonymous (2006) Agency for Toxic Substances and Disease Registry. 2006. CERCLA priority list of hazardous compounds.Washington, DC. http://www.atsdr.cdc.gov/cercla/05list.html

  • Becher D, Specht M, Hammer E, Francke W, Schauer F (2000) Cometabolic degradation of dibenzofuran by biphenyl-cultivated Ralstonia sp. strain SBUG 290. Appl Environ Microbiol 66:4528–4531

    Article  CAS  Google Scholar 

  • Borneman JP, Skroch PW, O’Sullivan PJA, Rumjanek NG, Jansen JL, Nienhuis J, Triplett EW (1996) Molecular microbial diversity of an agricultural soil in Wisconsin. Appl Environ Microbiol 62:1935–1943

    CAS  Google Scholar 

  • Burkhard J, Mackova M, Macek T, Kucerova P, Demnerova K (1997) Analytical procedure for the estimation of PCB transformation by plants. Anal Commun Roy Soc 34:287–290

    Article  CAS  Google Scholar 

  • Cebron AL, Bodrossy N, Stralis-Pavese AC, Singer IP, Thompson JI, Prosser IJ, Murrell JC (2007) Nutrient amendments in soil DNA stable isotope probing experiments reduce the observed methanotroph diversity. Appl Environ Microbiol 73:798–807

    Article  CAS  Google Scholar 

  • Chekol T, Vough LR, Chaney RL (2004) Phytoremediation of polychlorinated biphenyl-contaminated soils: the rhizosphere effect. Environ Int 30:799–804

    Article  CAS  Google Scholar 

  • Chroma L, Mackova M, Kucerova P, in der Wiesche C, Burkhard J, Macek T, Chroma L, Mackova M, Kucerova P, in der Wiesche C, Burkhard J, Macek T (2002a) Enzymes in plant metabolism of PCBs and PAHs. Acta Biotechnologica 22:35–41

    Article  CAS  Google Scholar 

  • Chroma L, Mackova M, Demnerova K, Macek T (2002b) Decolorization of RBBR by plant cells and correlation with the transformation of PCBs. Chemosphere 49:739–748

    Article  CAS  Google Scholar 

  • Cunningham SD, Berti WR (1993) Remediation of contaminated soils with green plants: an overview. In Vitro Cell Dev Biol 29:207–212

    Article  Google Scholar 

  • de Carcer DA, Martin M, Mackova M, Macek T, Karlson U, Rivilla R (2007) The introduction of genetically modified microorganisms designed for rhizoremediation induces changes on native bacteria in the rhizosphere but not in the surrounding soil. ISME J 1:215–223

    Article  CAS  Google Scholar 

  • Demnerova K, Mackova M, Spevakova V, Beranova K, Kochankova L, Lovecka P, Ryslava E, Macek T (2005) Two approaches to biological decontamination of ground-water and soil polluted by aromatics-characterization of microbial populations. Int Microbiol 8:205–211

    CAS  Google Scholar 

  • Doran PM (ed) (1997) Hairy roots: culture and application. Harwood Academic Publishers, London, pp 133–138

    Google Scholar 

  • Doran PM (2009) Application of plant tissue cultures in phytoremediation research: incentives and limitations. Biotechnol Bioeng 103:60–76

    Article  CAS  Google Scholar 

  • Doty SL, James CA, Moore AL, Vajzovic A, Singleton GL, Ma C, Khan Z, Xin G, Kang JW, Park AY et al (2007) Enhanced phytoremediation of volatile environmental pollutants with transgenic trees. Proc Natl Acad Sci USA 104:16816–16821

    Article  CAS  Google Scholar 

  • Dowling DN, Doty SL (2009) Improving phytoremediation through biotechnology. Curr Opin Biotechnol 20:1–3

    Article  CAS  Google Scholar 

  • Erb RW, Wagner-Dobler I (1993) Detection of polychlorinated biphenyl degradation genes in polluted sediments by direct DNA extraction and polymerase chain reaction. Appl Environ Microbiol 59:4065–4073

    CAS  Google Scholar 

  • Fletcher JS, Donnelly PK, Hegde RS (1995) Biostimulation of PCB-degrading bacteria by compounds released from plant roots. In: Hinchee RE, Anderson DB, Hoeppel RE (eds) Bioremediation of recalcitrant organics. Battelle Press, Columbus, pp 131–136

    Google Scholar 

  • Gibson DT, Cruden DL, Haddock JD, Zylstra GJ, Brand JM (1993) Oxidation of polychlorinated biphenyls by Pseudomonas sp. LB400 and Pseudomonas pseudoalcaligenes KF707. J Bacteriol 175:4561–4564

    CAS  Google Scholar 

  • Gichner T, Lovecka P, Kochankova L, Mackova M, Demnerova K (2007) Monitoring toxicity, DNA damage, and somatic mutations in tobacco plants growing in soil heavily polluted with polychlorinated biphenyls. Mutat Res 629:1–6

    CAS  Google Scholar 

  • Griga M, Bjelkova M, Tejklova E (2003) Phytoextraction of heavy metals by fibre crops: Linum usitatissimum L. case study. In: Kalogerakis N, Psillakis E (eds) Proceedings of the 2nd European Bioremediation Conference, Chania, Crete, TU Crete, pp 353–356

  • Hernandez BS, Koh SC, Chial M, Focht DD (1997) Terpene-utilizing isolates and their relevance to enhanced biotransformation of polychlorinated biphenyls in soil. Biodegradation 8:153–158

    Article  CAS  Google Scholar 

  • Holoubek I, Kocan A, Holoubkova I, Kohoutek J, Falandysz J, Roots O, Staffova K (2001) Polychlorinated biphenyls contaminated sites worldwide. The case of the Central and Eastern European countries. In: Robertson LW, Hansen LG (eds) PCBs. Recent advances in environmental, toxicology and health effects. The University Press of Kentucky, Lexington

    Google Scholar 

  • Ionescu M, Beranova K, Dudkova V, Kochankova L, Demnerova K, Macek T, Mackova M (2009) Isolation and characterization of different plant associated bacteria and their potential to degrade polychlorinated biphenyls. Int Biodeterior Biodegrad 63:667–672

    Article  CAS  Google Scholar 

  • Jurcek O, Wimmerova M, Wimmer Z (2008) Selected chiral alcohols: enzymic resolution and reduction of convenient substrates. Coord Chem Rev 252:767–781

    Article  CAS  Google Scholar 

  • Kas J, Burkhard J, Demnerova K, Kostal J, Macek T, Mackova M, Pazlarova J (1997) Perspectives in biodegradation of alkanes and PCBs. Pure Appl Chem 69:2357–2369

    Article  CAS  Google Scholar 

  • Kucerova P, Mackova M, Polachova L, Burkhard J, Demnerova K, Pazlarova J, Macek T (1999) Correlation of PCB transformation by plant tissue cultures with their morphology and peroxidase activity changes. Coll Czech Chem Commun 64:1497–1509

    Article  CAS  Google Scholar 

  • Kucerova P, Mackova M, Chroma L, Burkhard J, Triska J, Demnerova K, Macek T (2000) Metabolism of polychlorinated biphenyls by Solanum nigrum hairy root clone SNC-9O and analysis of transformation products. Plant Soil 225:109–115

    Article  CAS  Google Scholar 

  • Leigh MB, Prouzova P, Mackova M, Macek T, Nagle DP, Fletcher JS (2006) Polychlorinated biphenyl (PCB)-degrading bacteria associated with trees in a PCB-contaminated site. Appl Environ Microbiol 72:2331–2342

    Article  CAS  Google Scholar 

  • Leigh MB, Pellizari VH, Uhlik O, Sutka R, Rodrigues J, Ostrom NE, Zhou J, Tiedje JM (2007) Biphenyl-utilizing bacteria and their functional genes in a pine root zone contaminated with polychlorinated biphenyls (PCBs). ISME J 1:134–148

    Article  CAS  Google Scholar 

  • Liu W, Marsh T, Cheng H, Forney L (1997) Characterization of microbial diversity by determining terminal restriction fragment length polymorphisms of genes encoding 16S rRNA. Appl Environ Microbiol 63:4516–4522

    CAS  Google Scholar 

  • Macek T, Kotrba P, Suchova M, Skacel F, Demnerova K, Ruml T (1994) Accumulation of cadmium by hairy root cultures. Biotechnol Lett 16:621–624

    Article  CAS  Google Scholar 

  • Macek T, Mackova M, Burkhard J, Demnerova K (1998) Introduction of green plants for the control of metals and organics in environmental remediation. In: Holm FW (ed) Effluents from alternative demilitarization technologies. NATO PS Series 1, vol. 12. Kluwer Academic Publishers, Dordrecht, Boston, pp 71–85

    Google Scholar 

  • Macek T, Mackova M, Kas J (2000) Exploitation of plants for the removal of organics in environmental remediation. Biotechnol Advance 18:23–35

    Article  CAS  Google Scholar 

  • Macek T, Kotrba P, Svatos A, Novakova M, Demnerova K, Mackova M (2008) Novel roles for genetically modified plants in environmental protection. Trends Biotechnol 26:146–152

    Article  CAS  Google Scholar 

  • Mackova M, Macek T, Burkhard J, Ocenaskova J, Demnerova K, Pazlarova J (1997a) Biodegradation of polychlorinated biphenyls by plant cells. Int Biodeterior Biodegrad 39:317–325

    Article  CAS  Google Scholar 

  • Mackova M, Macek T, Kucerova P, Burkhard J, Pazlarova J, Demnerova K (1997b) Degradation of polychlorinated biphenyls by hairy root culture of Solanum nigrum. Biotechnol Let 9:787–790

    Article  Google Scholar 

  • Mackova M, Dowling D, Macek T (eds) (2006) Phytoremediation and rhizoremediation. Theoretical background. FOCUS on Biotechnology series, vol. 9A. Springer, Dordrecht, p 300

    Google Scholar 

  • Mackova M, Vrchotova B, Francova K, Sylvestre M, Tomaniova M, Lovecka P, Demnerova K, Macek T (2007) Biotransformation of PCBs by plants and bacteria, consequences of plant–microbe interactions. Eur J Soil Biol 43:233–241

    Article  CAS  Google Scholar 

  • McCutcheon SC, Schnoor JL (eds) (2003) Phytoremediation: transformation and control of contaminants. Wiley, New York

    Google Scholar 

  • McGuiness M, Dowling D (2009) Plant-associated bacterial degradation of toxic organic compounds in soil. Int J Environ Res Public Health. doi:10.3390/ijerph60x000x

    Google Scholar 

  • Narasimhan KC, Basheer VB, Bajic B, Swarup S (2003) Enhancement of plant-microbe interactions using a rhizosphere metabolomics-driven approach and its application in the removal of polychlorinated biphenyls. Plant Physiol 132:146–153

    Article  CAS  Google Scholar 

  • Novakova M, Mackova M, Chrastilova Z, Szekeres M, Demnerova K, Macek T (2009) Cloning the bacterial bphC gene into Nicotiana tabacum to improve the efficiency of PCB phytoremediation. Biotechnol Bioeng 102:29–37

    Article  CAS  Google Scholar 

  • Pavlikova D, Macek T, Mackova M, Pavlik M (2007) Monitoring native vegetation on a dumpsite of PCB-contaminated soil. Int J Phytoremed 9:71–78

    Article  CAS  Google Scholar 

  • Rezek J, Macek T, Mackova M, Triska J (2007) Plant metabolites of polychlorinated biphenyls in hairy root culture of black nightshade Solanum nigrum SNC-9O. Chemosphere 69:1221–1227

    Article  CAS  Google Scholar 

  • Rezek J, Macek T, Mackova M, Triska J, Ruzickova K (2008) Hydroxy-PCBs, methoxy-PCBs and hydroxy-methoxy-PCBs: Metabolites of polychlorinated biphenyls formed in vitro by tobacco cells. Environ Sci Technol 42:5746–5751

    Article  CAS  Google Scholar 

  • Rezek J, in der Wiesche C, Mackova M, Zadrazil F, Macek T (2009) Biodegradation of PAHs in long-term contaminated soil cultivated with European white birch (Betula pendula) and red mulberry (Morus rubra) tree. Int J Phytorem 11:1–16

    Article  CAS  Google Scholar 

  • Ryslava E, Krejcik Z, Macek T, Novakova H, Demnerova K, Mackova M (2003) Study of PCB degradation in real contaminated soil. Fresen Environ Bull 12:296–301

    CAS  Google Scholar 

  • Safe SH (1994) Polychlorinated biphenyls (PCBs): environmental impact, biochemical and toxic responses, and implications for risk assessment. Crit Rev Toxicol 24:87–149

    Article  CAS  Google Scholar 

  • Singer AC (2006) The chemical ecology of pollutant biodegradation. Bioremediation and phytoremediation from mechanistic and ecological perspectives. In: Mackova M, Dowling D, Macek T (eds) Phytoremediation and rhizoremediation. Theoretical background. Focus on biotechnology, 9A. Springer, Dordrecht, pp 5–21

    Chapter  Google Scholar 

  • Singer AC, Crowley DE, Thompson IP (2003) Secondary plant metabolites in phytoremediation and biotransformation. Trends Biotechnol 21:123–130

    Article  CAS  Google Scholar 

  • Soudek P, Podlipna R, Vanek T (1998) Phytoremediation of heavy metals by hairy root culture of Armoracia rusticana. Int J Biodeterior Biodegrad 42:235–236

    Google Scholar 

  • Sylvestre M (1980) Isolation method for bacterial isolates capable of growth on p-chlorobiphenyl. Appl Environ Microbiol 39:1223–1224

    CAS  Google Scholar 

  • Sylvestre M, Macek T, Mackova M (2009) Transgenic plants to improve rhizoremediation of polychlorinated biphenyls (PCBs). Curr Opin Biotechnol 20:242–247

    Article  CAS  Google Scholar 

  • Tillmann S, Strompl C, Timmis KN, Abraham WR (2005) Stable isotope probing reveals the dominant role of Burkholderia species in aerobic degradation of PCBs. FEMS Microbiol Ecol 52:207–217

    Article  CAS  Google Scholar 

  • Tlustos P, Pavlikova D, Szakova J, Fischerova Z, Balik J (2006) Exploitation of fast growing trees in metal remediation. In: Mackova M, Dowling D, Macek T (eds) Phytoremediation and rhizoremediation. Theoretical background. Focus on biotechnology series, vol. 9A. Springer, Dordrecht

    Google Scholar 

  • Uhlik O, Jecna K, Leigh MB, Mackova M, Macek T (2009a) DNA-based stable isotope probing: a link between community structure and function. Sci Total Environ 407:3611–3619

    Article  CAS  Google Scholar 

  • Uhlik O, Jecna K, Mackova M, Vlcek C, Hroudova M, Demnerova K, Paces V, Macek T (2009b) Biphenyl-metabolizing bacteria in the rhizosphere of horseradish and bulk soil contaminated by polychlorinated biphenyls as revealed by stable isotope probing. Appl Environ Microbiol. doi:10.1128/AEM.00466-09

    Google Scholar 

  • Van Aken B (2008) Transgenic plants for phytoremediation: helping nature to clean up environmental pollution. Trends Biotechnol 26:225–227

    Article  CAS  Google Scholar 

  • Villacieros M, Whelan CM, Mackova M, Molgaard J, Sanchez-Contreras M, Lloret J, Aguirre de Carcer D, Bolanos L, Oruezabal RI, Macek T, Karlson U, Dowling DN, Martin M, Rivilla R (2005) PCB rhizoremediation by Pseudomonas fluorescens F113 derivatives using a Sinorhizobium meliloti nod system to drive bph gene expression. Appl Environ Microbiol 71:2687–2694

    Article  CAS  Google Scholar 

  • Weyens N, van der Lelie D, Taghavi S, Vangronsveld J (2009) Phytoremediation: plant–endophyte partnerships take the challenge. Curr Opin Biotechnol 20:248–254

    Article  CAS  Google Scholar 

  • Wilken A, Bock C, Bokern M, Harms H (1995) Metabolism of different PCB congeners in plant cell cultures. Environ Chem Toxicol 14:2017–2022

    Article  CAS  Google Scholar 

  • Zarevucka M, Wimmer Z (2008) Plant products for pharmacology: application of enzymes in their transformations. Int J Mol Sci 9:2447–2473

    Article  CAS  Google Scholar 

  • Zhou J, Bruns MA, Tiedje JM (1996) DNA recovery from soils of diverse composition. Appl Environ Microbiol 62:316–322

    CAS  Google Scholar 

Download references

Acknowledgements

The authors thank the support of projects COST 859, the Grant Agency of the Czech Republic 525/09/1058, and projects funded by Czech Ministry of Education 2B06156, 6046137305, Z40550506. The help of many colleagues and students is highly appreciated. Especially, Mr. Rudolf Kaplanek deserves special thanks for his enthusiasm, patience and permission to access to site.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Tomas Macek.

Additional information

Responsible editors: Peter Schröder and Jean-Paul Schwitzguébel

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mackova, M., Prouzova, P., Stursa, P. et al. Phyto/rhizoremediation studies using long-term PCB-contaminated soil. Environ Sci Pollut Res 16, 817–829 (2009). https://doi.org/10.1007/s11356-009-0240-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11356-009-0240-3

Keywords

Navigation