Advertisement

Environmental Science and Pollution Research

, Volume 16, Issue 7, pp 830–843 | Cite as

Bacteria associated with oak and ash on a TCE-contaminated site: characterization of isolates with potential to avoid evapotranspiration of TCE

  • Nele Weyens
  • Safiyh Taghavi
  • Tanja Barac
  • Daniel van der Lelie
  • Jana Boulet
  • Tom Artois
  • Robert Carleer
  • Jaco Vangronsveld
COST ACTION 859 • PHYTOREMEDIATION • RESEARCH ARTICLE

Abstract

Background, aim, and scope

Along transects under a mixed woodland of English Oak (Quercus robur) and Common Ash (Fraxinus excelsior) growing on a trichloroethylene (TCE)-contaminated groundwater plume, sharp decreases in TCE concentrations were observed, while transects outside the planted area did not show this remarkable decrease. This suggested a possibly active role of the trees and their associated bacteria in the remediation process. Therefore, the cultivable bacterial communities associated with both tree species growing on this TCE-contaminated groundwater plume were investigated in order to assess the possibilities and practical aspects of using these common native tree species and their associated bacteria for phytoremediation. In this study, only the cultivable bacteria were characterized because the final aim was to isolate TCE-degrading, heavy metal resistant bacteria that might be used as traceable inocula to enhance bioremediation.

Materials and methods

Cultivable bacteria isolated from bulk soil, rhizosphere, root, stem, and leaf were genotypically characterized by amplified rDNA restriction analysis (ARDRA) of their 16S rRNA gene and identified by 16S rRNA gene sequencing. Bacteria that displayed distinct ARDRA patterns were screened for heavy metal resistance, as well as TCE tolerance and degradation, as preparation for possible future in situ inoculation experiments. Furthermore, in situ evapotranspiration measurements were performed to investigate if the degradation capacity of the associated bacteria is enough to prevent TCE evapotranspiration to the air.

Results and discussion

Between both tree species, the associated populations of cultivable bacteria clearly differed in composition. In English Oak, more species-specific, most likely obligate endophytes were found. The majority of the isolated bacteria showed increased tolerance to TCE, and TCE degradation capacity was observed in some of the strains. However, in situ evapotranspiration measurements revealed that a significant amount of TCE and its metabolites was evaporating through the leaves to the atmosphere.

Conclusions and perspectives

The characterization of the isolates obtained in this study shows that the bacterial community associated with Oak and Ash on a TCE-contaminated site, was strongly enriched with TCE-tolerant strains. However, this was not sufficient to degrade all TCE before it reaches the leaves. A possible strategy to overcome this evapotranspiration to the atmosphere is to enrich the plant-associated TCE-degrading bacteria by in situ inoculation with endophytic strains capable of degrading TCE.

Keywords

Chlorinated solvents Fraxinus excelsior Phytoremediation Plant-associated bacteria Quercus robur TCE Evapotranspiration Endophytes 

Notes

Acknowledgements

This research was funded by the Institute for the Promotion of Innovation through Science and Technology in Flanders (IWT-Vlaanderen) for N.W. and by the Fund for Scientific Research Flanders (FWO-Vlaanderen), Ph.D. grant for J.B. and postdoc grant for T.B. This work was also supported by the UHasselt Methusalem project 08M03VGRJ. D.v.d.L. and S.T. are supported by the US Department of Energy, Office of Science, BER, project number KP1102010 under contract DE-AC02-98CH10886, and by Laboratory Directed Research and Development funds (LDRD05-063) at the Brookhaven National Laboratory under contract with the U.S. Department of Energy. We thank A. Wijgaerts and C. Put for their help with the isolation and J. Put, J. Czech, and R. Carleer for GC analysis.

References

  1. Arshad M, Frankenberger WT (1991) Microbial production of plant hormones. Kluwer Academic Publishers, Dordrecht, the NetherlandsGoogle Scholar
  2. Barac T, Taghavi S, Borremans B, Provoost A, Oeyen L, Colpaert JV, Vangronsveld J, van der Lelie D (2004) Engineered endophytic bacteria improve phytoremediation of water-soluble, volatile, organic pollutants. Nat Biotechnol 22:583–588CrossRefGoogle Scholar
  3. Barac T, Weyens N, Oeyen L, Taghavi S, van der Lelie D, Dubin D, Spliet M, Vangronsveld J (2009) Field note: hydraulic containment of a BTEX plume using poplar trees. Int J Phytorem 11:416–424CrossRefGoogle Scholar
  4. Bron S, Venema G (1972) Ultraviolet inactivation and excision-repair in Bacillus subtilis. I. Construction and characterization of a transformable eightfold auxotrophic strain and two ultraviolet-sensitive derivatives. Mut Res 15:1–10Google Scholar
  5. Burken JG, Schnoor JL (1999) Distribution and volatilization of organic compounds following uptake by hybrid Poplar trees. Int J Phytorem 1:139–151CrossRefGoogle Scholar
  6. Burken JG, Ma X (2006) Phytoremediation of volatile organic compounds. In: Mackova M, Dowling DN, Macek T (eds), Phytoremediation and Rhizoremediation Theoretical Background. pp 199–216Google Scholar
  7. Cankar K, Kraigher H, Ravnikar M, Rupnik M (2005) Bacterial endophytes from seeds of Norway spruce (Picea abies L. Karts). FEMS Microbiol Lett 244:341–345CrossRefGoogle Scholar
  8. Coombs JT, Franco CMM (2003) Isolation and identification of actinobacteria from surface-sterilized wheat roots. Appl Environ Microbiol 69:5603–5608CrossRefGoogle Scholar
  9. Devers M, Henry S, Hartmann A, Martin-Laurent F (2005) Horizontal gene transfer of atrazine-degrading genes (atz) from Agrobacterium tumefaciens St96-4pADPA:Tn5 to bacteria of maize-cultivated soil. Pest Manag Sci 61:870–880CrossRefGoogle Scholar
  10. Döbereiner J, Urquiaga S, Boddey RM (1995) Alternatives for nitrogen nutrition of crops in tropical agriculture. Fert Res 42:339–346CrossRefGoogle Scholar
  11. Dong Q, Springael D, Schoeters J, Nuyts G, Mergeay M, Diels L (1998) Horizontal transfer of bacterial heavy metal resistance genes and its applications in activated sludge systems. Water Sci Technol 37:465–468CrossRefGoogle Scholar
  12. Doucette WJ, Bugbee B, Hayhurst S, Plaehn WA, Downey DC, Taffinder SA, Edwards R (1998) Phytoremediation of dissolved phase trichloroethylene using mature vegetation. In: Wickramanayake GB, Hinchee HE (eds) Bioremediation and Phytoremediation: Chlorinated and Recalcitrant Compounds. Batelle Press, Columbus, USA, pp 251–256Google Scholar
  13. Ferro AM, Rieder JP, Kennedy J, Kjelgren R (1997) Phytoremediation of groundwater using Poplar trees. In: Thibault CA (ed) Phytoremediation. Inc, International Business Communications, pp 202–212Google Scholar
  14. Germaine K, Keogh E, Borremans B, van der Lelie D, Barac T, Oeyen L, Vangronsveld J, Porteus Moore F, Moore ERB, Campbel CD, Ryan D, Dowling D (2004) Colonisation of Poplar trees by gfp expressing endophytes. FEMS Microbiol Ecol 48:109–118CrossRefGoogle Scholar
  15. Glick BR (2004) Bacterial ACC deaminase and the alleviation of plant stress. Adv Appl Microbiol 56:291–312CrossRefGoogle Scholar
  16. Glick BR, Jacobson CB, Schwarze MK, Pasternak JJ (1994) 1-Aminocyclopropane-1-carboxylic acid deaminase mutants of the plant growth promoting rhizobacterium Pseudomonas putida GR 12–2 do not stimulate canola root elongation. Can J Microbiol 40:911–915CrossRefGoogle Scholar
  17. Glick BR, Penrose DM, Li J (1998) A model for the lowering of plant ethylene concentrations by plant growth promoting bacteria. J Theor Biol 190:63–68CrossRefGoogle Scholar
  18. Hebbar KP, Davey AG, Merrin J, Dart PJ (1992a) Rhizobacteria of maize antagonistic to Fusarium moniliforme, a soil-borne fungal pathogen: colonization of rhizosphere and roots. Soil Biol Biochem 24:989–997CrossRefGoogle Scholar
  19. Hebbar KP, Davey AG, Merrin J, McLoughlin TJ, Dart PJ (1992b) Pseudomonas cepacia, a potential suppressor of maize soil-borne diseases-seed inoculation and maize root colonization. Soil Biol Biochem 24:999–1007CrossRefGoogle Scholar
  20. Kuklinsky-Sobral J, Araujo WL, Mendes R, Geraldi IO, Pizzirani-Kleiner AA, Azevedo JL (2004) Isolation and characterization of soybean-associated bacteria and their potential for plant growth promotion. Environ Microbiol 6:1244–51CrossRefGoogle Scholar
  21. Lodewyckx C, Vangronsveld J, Porteous F, Moore ERB, Taghavi S, Mergeay M, van der Lelie D (2002) Endophytic bacteria and their potential applications. Critical Rev Plant Sci 21:583–606CrossRefGoogle Scholar
  22. Ma X, Burken JG (2003) TCE diffusion to the atmosphere in phytoremediation applications. Environ Sci Technol 37:2534–2539CrossRefGoogle Scholar
  23. Mars AE, Houwing J, Dolfing J, Janssen DB (1996) Degradation of toluene and trichloroethylene by Burkholderia cepacia G4 in growth-limited fed-batch culture. Appl Environ Microbiol 62:886–891Google Scholar
  24. Mastretta C, Barac T, Vangronsveld J, Newman L, Taghavi S, van der Lelie D (2006) Endophytic bacteria and their potential application to improve the phytoremediation of contaminated environments. Biotech Gen Eng Rev 23:175–207Google Scholar
  25. Mergeay M, Nies D, Schlegel HG, Gerits J, Charles P, Van Gijsegem F (1985) Alcaligenes eutrophus CH34 is a facultative chemolithotroph with plasmid-bound resistance to heavy metals. J Bacteriol 162:328–334Google Scholar
  26. Newman LA, Reynolds CM (2005) Bacteria and phytoremediation: new uses for endophytic bacteria in plants. Trends Biotechnol 23:6–8CrossRefGoogle Scholar
  27. Porteous Moore F, Barac T, Borremans B, Oeyen L, Vangronsveld J, van der Lelie D, Campbell CD, Moore ERB (2006) Endophytic bacterial diversity in Poplar trees growing on a BTEX-contaminated site: the characterisation of isolates with potential to enhance phytoremediation. Sys App Micro 29:539–556CrossRefGoogle Scholar
  28. Ronchel MC, Ramos-Diaz MA, Ramos JL (2000) Retrotransfer of DNA in the rhizosphere. Environ Microbiol 2:319–323CrossRefGoogle Scholar
  29. Ryoo D, Shim H, Canada K, Barbieri P, Wood TK (2000) Aerobic degradation of tetrachloroethylene by toluene-o-xylene monooxygenase of Pseudomonas stutzeri OX1. Nat Biotechnol 18:775–778CrossRefGoogle Scholar
  30. Ryoo D, Shim H, Arenghi FLG, Barbieri P, Wood TK (2001) Tetrachloroethylene, trichloroethylene, and chlorinated phenols induce toluene-o-monooxygenase activity in Pseudomonas stutzeri OX1. Appl Microbiol Biotechnol 56:545–549CrossRefGoogle Scholar
  31. Ryu CM, Farag MA, Hu CH, Reddy MS, Wei HX, Pare PW, Kloepper JW (2003) Bacterial volatiles promote growth in Arabidopsis. Proc Natl Acad Sci USA 100:4927–4932CrossRefGoogle Scholar
  32. Schatz A, Bovell C (1952) Growth and hydrogenase activity of a new bacterium Hydrogenomonas facilis. J Bacteriol 63:87–98Google Scholar
  33. Schnoor LJ, Licht AL, McCutchon CS, Wolfe NL, Carreira HL (1995) Phytoremediation of organic and nutrient contaminants. Environ Sci Technol 29:318A–323ACrossRefGoogle Scholar
  34. Schwitzguébel JP, van der Lelie D, Glass DJ, Vangonsveld J, Baker AJM (2002) Phytoremediation: European and American trends, successes, obstacles and needs. J Soils Sediments 2:91–99CrossRefGoogle Scholar
  35. Shim H, Chauhan S, Ryoo D, Bowers K, Thomas SM, Canada KA, Burken J, Wood TK (2000) Rhizosphere competitiveness of trichloroethylene-degrading, poplar colonizing recombinant bacteria. Appl Environ Microbiol 66:4673–4678CrossRefGoogle Scholar
  36. Shim H, Ryoo D, Barbieri P, Wood TK (2001) Aerobic degradation of mixtures of tetrachloroethylene, trichloroethylene, dichloroethylenes, and vinyl chloride by toluene-o-xylene monooxygenase of Pseudomonas stutzeri OX1. Appl Microbiol Biotechnol 56:265–269CrossRefGoogle Scholar
  37. Siciliano SD, Fortin N, Mihoc A, Wisse G, Labelle S, Beaumier D, Ouellette D, Roy R, Whyte LG, Banks MK, Schwab P, Lee K, Greer CW (2001) Selection of specific endophytic bacterial genotypes by plants in response to soil contamination. Appl Environ Microbiol 67:2469–2475CrossRefGoogle Scholar
  38. Swofford DL (2003) PAUP*—Phylogenetic Analysis Using Parsimony (*and Other Methods), Ver. 4. [Computer Software and Manual.]. Sinauer Associates, Sunderland, MAGoogle Scholar
  39. Taghavi S, Barac T, Greenberg B, Borremans B, Vangronsveld J, van der Lelie D (2005) Horizontal gene transfer to endogenous endophytic bacteria from Poplar improves phytoremediation of toluene. Appl Environ Microbiol 71:8500–8505CrossRefGoogle Scholar
  40. Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The CLUSTAL_X windows interface: flexible strategies for multiple sequences aided by quality analysis tools. Nucleic Acid Res 25:4876–4882CrossRefGoogle Scholar
  41. Top EM, Springael D, Boon N (2002) Catabolic mobile genetic elements and their potential use in bioaugmentation of polluted soils and waters. FEMS Microbiol Ecol 42:199–208CrossRefGoogle Scholar
  42. Trapp S, Zambrano KC, Kusk KC, Karlson U (2000) A phytotoxicity test using transpiration of willows. Arch Env Contam Toxicol 39:154–160CrossRefGoogle Scholar
  43. Triplett EW (1996) Diazotrophic endophytes: progress and prospects for nitrogen fixation in monocots. Plant Soil 186:29–38CrossRefGoogle Scholar
  44. van der Lelie D, Schwitzguébel JP, Vangronsveld J, Baker AJM (2001) Assessing phytoremediation’s progress in the United States and Europe. Environ Sci Technol 35:446A–452ACrossRefGoogle Scholar
  45. Van Elsas JD, Gardener BB, Wolters AC, Smit E (1998) Isolation, characterization, and transfer of cryptic gene-mobilizing plasmids in the wheat rhizosphere. Appl Environ Microbiol 64:880–889Google Scholar
  46. Yee DC, Maynard JA, Wood TK (1998) Rhizoremediation of trichloroethylene by a recombinant, root-colonizing Pseudomonas fluorescens strain expressing toluene ortho-monooxygenase constitutively. Appl Environ Microbiol 64:112–118Google Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Nele Weyens
    • 1
  • Safiyh Taghavi
    • 4
  • Tanja Barac
    • 1
  • Daniel van der Lelie
    • 4
  • Jana Boulet
    • 1
  • Tom Artois
    • 2
  • Robert Carleer
    • 3
  • Jaco Vangronsveld
    • 1
  1. 1.Environmental BiologyHasselt UniversityDiepenbeekBelgium
  2. 2.Biodiversity, Phylogeny and Population StudiesHasselt UniversityDiepenbeekBelgium
  3. 3.Applied ChemistryHasselt UniversityDiepenbeekBelgium
  4. 4.Biology DepartmentBrookhaven National Laboratory (BNL)UptonUSA

Personalised recommendations