[go: up one dir, main page]
Skip to main content

Advertisement

SpringerLink
Log in

Polychlorinated biphenyl (PCB) degradation and persistence of a gfp-marked Ralstonia eutropha H850 in PCB-contaminated soil

  • Original Paper
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

Ralstonia eutropha H850 was labelled chromosomally with a gfp marker gene encoding for the green fluorescent protein, and designated R. eutropha H850g13. Visual observation of green fluorescent cells under an epifluorescence microscope, and PCR amplification products, confirmed that the bacterium was labelled with gfp. Southern blot hybridization products further confirmed the gfp was chromosomally labelled. Using resting cell assays, it was determined that insertion of the gfp gene decreased the microorganisms' ability to degrade biphenyl compared to the parent strain. However, this marker facilitated the identification and monitoring of R. eutropha H850g13 survival in soil microcosm experiments. Survival and polychlorinated biphenyl degradation by R. eutropha H850g13 was analysed in soil microcosms spiked with 2,2′,5,5′-tetrachlorobiphenyl (TeCB). R. eutropha H850g13 was detected by viable plate counts and most-probable-number/PCR after 102 days in TeCB-contaminated soil microcosms, and was likely outcompeted by indigenous soil microorganisms in microcosms amended with oil and Daramend (an organic amendment, http://www.adventusremediation.com). R. eutropha H850g13 did not degrade TeCB in any of the soil microcosms. This research confirmed that gfp was useful as a marker to distinguish R. eutropha H850g13 from indigenous soil microorganisms over a 102 day period and that, under the experimental conditions used, R. eutropha H850g13 did not degrade TeCB.

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

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1.
Fig. 2A–D.
Fig. 3.
Fig. 4.
Fig. 5.

Similar content being viewed by others

Explore related subjects

Discover the latest articles, news and stories from top researchers in related subjects.

References

  • Ahn Y-B, Beaudette LA, Lee H, Trevors JT (2000) Survival of a GFP-labeled polychlorinated biphenyl degrading psychrotolerant Pseudomonas spp. in 4 and 22°C soil microcosms. Microb Ecol 42:614–623

    Article  Google Scholar 

  • Andersen JB, Sternberg C, Poulsen LK, Bjorn SP, Givskov M, Molin S (1998) New unstable variants of green fluorescent protein for studies of transient gene expression in bacteria. Appl Environ Microbiol 64:2240–2246

    CAS  PubMed  Google Scholar 

  • Aronstein BN, Paterek JR, Kelley RL, Rice LE (1995) The effect of chemical pretreatment on the aerobic microbial degradation of PCB congeners in aqueous systems. J Ind Microbiol 15:55–59

    CAS  PubMed  Google Scholar 

  • Ausubel FM, Brent R, Kingston RE, Moore DD, Serdman JG, Smith JA, Struhl K (eds) (1989) Short protocols in molecular biology. Wiley, New York

  • Bedard DL, Unterman R, Bopp LH, Brennan MJ, Haberl ML, Johnson C (1986) Rapid assay for screening and characterizing microorganisms for the ability to degrade polychlorinated biphenyls. Appl Environ Microbiol 51:761–768

    CAS  PubMed  Google Scholar 

  • Bedard DL, Haberl ML, May RJ, Brennan M (1987) Evidence for novel mechanisms of polychlorinated biphenyl metabolism in Alcaligenes eutrophus H850. Appl Environ Microbiol 53:1103–1112

    CAS  PubMed  Google Scholar 

  • Billingsley KA, Backus SM, Juneson C, Ward OP (1997a) Comparison of the degradation patterns of polychlorinated biphenyl congeners in Aroclors by Pseudomonas strain LB400 after growth on various carbon sources. Can J Microbiol 43:1172–1179

    CAS  PubMed  Google Scholar 

  • Billingsley KA, Backus SM, Ward OP (1997b) Studies on the transformation of selected polychlorinated biphenyl congeners by Pseudomonas strain LB400. Can J Microbiol 43:782–788

    CAS  Google Scholar 

  • Boehringer Mannheim (1995) The DIG system user's guide for filter hybridization. Biochemica, Boehringer Mannheim, Germany

  • Brock TD, Madigan MT, Martinko JM, Parker J (1994) Biology of microorganisms. Prentice Hall, New Jersey

  • Cassidy MB, Leung KT, Lee H, Trevors JT (2000) A comparison of enumeration methods for culturable Pseudomonas fluorescens cells marked with green fluorescent protein. J Microbiol Methods 40:135–145

    Article  CAS  PubMed  Google Scholar 

  • Celi L, Gennari M, Schnitzer M, Khan SU (1997). Extractable and nonextractable (bound) residues of acifluorfen in an organic soil. J Agric Food Chem 45:3677–3680

    Article  CAS  Google Scholar 

  • Dyke MI van, Lee H, Trevors JT (1996) Survival of lux AB-marked Alcaligenes eutrophus H850 in PCB-contaminated soil and sediment. J Chem Tech Biotechnol 65:115–122

    Article  Google Scholar 

  • Elsas JD van, Dijkstra AF, Govaert JM, van Veen JA (1986) Survival of Pseudomonas fluorescens and Bacillus subtilis introduced into two soils of different texture in field microplots. FEMS Microbiol Ecol 38:151–160

    Google Scholar 

  • Elsas JD van, Waalwijk C (1991) Methods for the detection of specific bacteria and their genes in soil. Agric Ecosyst Environ 34:97–105

    Google Scholar 

  • England LS, Lee H, Trevors JT (1993) Bacterial survival in soil: effect of clays and protozoa. Soil Biol Biochem 25:525–531

    Article  Google Scholar 

  • Errampalli D, Trevors JT, Lee H, Leung K, Cassidy M, Knoke K, Marwood T, Shaw K, Blears M, Chung E (1997) Bioremediation: a perspective. J Soil Contam 6:207–218

    CAS  Google Scholar 

  • Errampalli D, Okamura H, Lee H, Trevors JT, van Elsas JD (1998) Green fluorescent protein as a marker to monitor survival of phenanthrene-mineralizing Pseudomonas sp. UG14Gr in creosote-contaminated soil. FEMS Microbiol Ecol 26:181–191

    CAS  Google Scholar 

  • Flemming CA, Leung KT, Lee H, Trevors JT (1994) Survival of lux-lac-marked biosurfactant-producing Pseudomonas aeruginosa UG2L in soil monitored by non-selective plating and PCR. Appl Environ Microbiol 60:1606–1613

    CAS  PubMed  Google Scholar 

  • Furukawa K (1994) Genetic systems in soil bacteria for the degradation of polychlorinated biphenyls. In: Chaudry GR (ed) Biological degradation and bioremediation of toxic chemicals. Dioscorides Press, Portland, Ore., pp 33–46

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

    CAS  PubMed  Google Scholar 

  • Haddock JD, Horton JR, Gibson DT (1995) Dihydroxylation and dechlorination of chlorinated biphenyls by purified biphenyl 2,3-dioxygenase from Pseudomonas sp. strain LB400. J Bacteriol 177:20–26

    CAS  PubMed  Google Scholar 

  • Hurley AH, Roscoe ME (1983) Automated statistical analysis of microbial enumeration by dilution series. J Appl Bacteriol 55:159–164

    Google Scholar 

  • Leung KT, Watt A, Lee H, Trevors JT (1997) Quantitative detection of pentachlorophenol-degrading Sphingomonas sp. UG30 in soil by a most-probable-number/polymerase chain reaction protocol. J Microbiol Methods 31:59–66

    Article  Google Scholar 

  • Leveau JHJ, van der Meer JR (1996) The tfdR gene product can successfully take over the role of the insertion element-inactivated TfdT protein as a transcriptional activator of the tfdCDEF gene cluster, which encodes chlorocatechol degradation in Ralstonia eutropha JMP134(pJP4). J Bacteriol 178:6824–6832

    CAS  PubMed  Google Scholar 

  • Middeldorp PJM, Briglia M, Salkinoja-Salonen MS (1990) Biodegradation of pentachlorophenol in natural soil by inoculated Rhodococus chlorophenolicus. Microb Ecol 20:129–139

    Google Scholar 

  • Overbeek LS van, Eberl L, Givskov M, Molin S, van Elsas JD (1995) Survival of, and induced stress resistance in, carbon-starved Pseudomonas fluorescens cells residing in soil. Appl Environ Microbiol 61:4202–4208

    PubMed  Google Scholar 

  • Ripp S, Nivens DE, Werner C, Sayler GS (2000) Bioluminescent most-probable-number monitoring of a genetically engineered bacterium during a long-term contained field release. Appl Microbiol Biotechnol 53:736–741

    Article  CAS  PubMed  Google Scholar 

  • Seech AG, Trevors JT, Bulman TL (1990) Biodegradation of pentachlorophenol in soil: the response to physical, chemical, and biological treatments. Can J Microbiol 37:440–444

    Google Scholar 

  • Seech AG, Cairns JE, Marvan IJ (2000) Composition and method for dehalogenation and degradation of halogenated organic contaminants. US Patent 6,083,394. Grace of Canada, Mississauga, Canada

  • So JS, Chang WS, Leung KT, Lee H, Trevors JT (2000) Simultaneous detection of gfp-marked Moraxella sp. G21r and lux-marked Ralstonia eutropha H850Lr using most-probable-number method. J Microbiol Methods 43:67–72

    Article  CAS  PubMed  Google Scholar 

  • Tsien R (1998) The green fluorescent protein. Annu Rev Biochem 67:509–544

    CAS  PubMed  Google Scholar 

  • Veen JA van, van Overbeek LS, van Elsas JD (1997) Fate and activity of microorganisms introduced into soil. Microbiol Mol Biol Rev 61:121–135

    PubMed  Google Scholar 

  • Yabuuchi E, Kosako Y, Yano I, Hotta H, Nishiuchi Y (1995) Transfer of two Burkholderia and an Alcaligenes species to Ralstonia gen. nov.: proposal of Ralstonia pickettii (Ralston, Palleroni and Doudoroff 1973) comb. nov., Ralstonia solanacearum (Smith 1896) comb. nov. and Ralstonia eutropha (Davis 1969) comb. nov. Microbiol Immunol 39:897–904

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by a Natural Sciences and Engineering Research Strategic and Discovery Grants to J.T. Trevors and H. Lee. We would also like to thank our industrial collaborator Dr. A. Seech, Grace Bioremediation Technologies, Mississauga, Canada, and Dr. S. Molin, Technical University of Denmark, Denmark for providing the pJBA29.

Author information

Authors and Affiliations

  1. Department of Environmental Biology, University of Guelph, Guelph, ON, N1G 2W1, Canada

    A.-M. Irwin Abbey, H. Lee & J. T. Trevors

  2. Wastewater Technology Centre, Environment Canada, Burlington, ON, L7R 4A6, Canada

    L. A. Beaudette

Authors
  1. A.-M. Irwin Abbey
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. L. A. Beaudette
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. H. Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. J. T. Trevors
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to J. T. Trevors.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Irwin Abbey, AM., Beaudette, L.A., Lee, H. et al. Polychlorinated biphenyl (PCB) degradation and persistence of a gfp-marked Ralstonia eutropha H850 in PCB-contaminated soil. Appl Microbiol Biotechnol 63, 222–230 (2003). https://doi.org/10.1007/s00253-003-1380-x

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00253-003-1380-x

Keywords

Search

Navigation