Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research
  • Published:

Survival, Colonization and Dispersal of Genetically Modified Pseudomonas fluorescens SBW25 in the Phytosphere of Field Grown Sugar Beet

Bio/Technology volume 13, pages 1493–1497 (1995)Cite this article

Abstract

A genetically modified microorganism (GMM), Pseudomonas fluorescens SBW25 (lacZY and kanr-xylE), derived from a strain taken from the leaves of mature sugar beet (Beta vulgaris var. Amethyst), was released as a seed inoculum (ca 1×107 cfu/seed) to field-grown sugar beet in 1993 and 1994. The inoculum successfully colonized the roots and leaves of developing plants and survived in the phytosphere throughout the growing season, (270 days post release). Variation in the population dynamics of the GMM colonizing the immature leaf habitat was observed over two consecutive seasons. In 1993, the GMM represented a minor component of the pseudomonad community in emerging seedlings (<6%), (4×104 cfu g−1, coefficient of variation, CV=7%) whereas, in 1994, plants of similar size were colonized by significantly greater numbers of the GMM (1.1 × 106 cfu g−1, CV=10%) (81% of the total pseudomon-ads) reaching maximum densities 102 days after release. Limited dispersal of the GMM from sugar beet to other plants species was recorded. Although GMMs could not be detected in soil (<1 cfu g−1) or on overwintering plant tissue (<20 cfu g−1) GMMs persisted and colonised plants grown the following season. No transfer of the marker genes to other indigenous microorganisms was recorded.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Similar content being viewed by others

References

  1. Wilson, M. and Lindow, S.E. 1993. Release of recombinant microorganisms. Annu. Rev. Microbiol. 47: 913–944.

    Article  CAS  PubMed  Google Scholar 

  2. Cory, J.S., Hirst, M.L., Williams, T., Hails, R.S., Goulson, D., Green, B.M., Carty, T.M., Possee, R.D., Cayley, J. and Bishop, D.H.L. 1994. Field trial of a genetically improved baculovirus insecticide. Nature 370: 138–140.

    Article  Google Scholar 

  3. Lindow, S.E., Knudsen, G.R., Seidler, R.J., Walter, M.V., Lambou, V.W., Amy, P.S., Schmedding Prince, V. and Hern, S. 1988. Aerial dispersal and epiphytic survival of Pseudomonas syringae. Appl. Eviron. Microbiol. 54: 1557–1563.

    CAS  Google Scholar 

  4. Drahos, D.J. 1991. Field testing of genetically engineered microorganisms. Advances in Biotechnology 9: 157–177.

    Article  CAS  Google Scholar 

  5. Tiedje, J.M., Colwell, R.K., Grossman, Y.L., Hodgson, R.E., Lenski, R.E., Mack, R.N. and Regal, P.J. 1989. The planned introduction of genetically modified organisms: ecological considerations and recommendations. Ecology 70: 298–315.

    Article  Google Scholar 

  6. Drahos, D.J., Barry, G.F., Hemmin, B.C., Brandt, E.J., Kline, E.L., Skipper, H.D., Kluepfel, D.A., Gooden, D.T. and Hughes, T.A. 1992. Spread and survival of genetically marked bacteria in soil, p. 147–159. In: Release of genetically engineered and other microorganisms, Fry, J. C. and Day, M. J. (Eds.). Cambridge University Press.

    Google Scholar 

  7. Kluepfel, D.A., Kline, E.L., Skipper, H.D., Hughes, T.A., Gooden, D.T., Barry, G.F., Hemmings, B.C. and Brandt, E.J. 1991. The release and tracking of genetically engineered bacteria in the environment. Phytopathology 81: 348–352.

    Google Scholar 

  8. Ryder, M. 1994. Key issues in the deliberate release of genetically manipulated bacteria. FEMS Microbiol. Ecol. 15: 139–146.

    Article  CAS  Google Scholar 

  9. Parke, J.L., Zablotowicz, R.M. and Rand, R.E. 1992. Tracking a lacZY marked strain of Pseudomonas fluorescens in a Wisconsin field trial. Phytopathology 82: 1177–1178.

    Google Scholar 

  10. Lindemann, J. and Suslow, T.V. 1987. Competition between ice nucleation active wild type and ice nucleation deficient deletion mutant strains of Pseudomonas syringae and Pseudomonas fluorescens biovar I and biological control of frost injury on strawberry blossoms. Phytopathology 77: 882–886.

    Article  Google Scholar 

  11. Shaw, J.J., Dane, F., Geiger, D. and Kloepper, J.W. 1992. Use of bioluminescence for detection of genetically engineered microorganisms released into the environment. Appl. Environ. Microbiol. 58: 267–273.

    CAS  PubMed  PubMed Central  Google Scholar 

  12. Hirano, S.S. and Upper, C.D. 1993. Dynamics, Spread, and persistence of a single genotype of Pseudomonas syringae relative to those of its conspecifics on populations on snap bean leaflets. Appl. Eviron. Microbiol. 59: 1082–1091.

    CAS  Google Scholar 

  13. Thompson, I.P., Bailey, M.J., Fenlon, J.S., Fermor, T.R., Lilley, A.K., Lynch, J.M., McCormack, P.J., McQuilken, M., Purdy, K.J., Rainey, P.B. and Whipps, J.M. 1993. Quantitative and qualitative seasonal changes in the microbial community from the phyllosphere of sugar beet (Beta vulgaris). Plant and Soil 150: 177–191.

    Article  Google Scholar 

  14. Thompson, I.P., Bailey, M.J., Ellis, R.J. and Purdy, K.J. 1993. Subgrouping of bacterial populations by cellular fatty acid composition. FEMS Microbiol. Ecol. 102: 75–84.

    Article  CAS  Google Scholar 

  15. Thompson, I.P., Bailey, M.J., Ellis, R.J., Lilley, A.K., McCormack, P.J., Purdy, K.J. and Rainey, P.B. 1995. Short term community dynamics in the phyllosphere Microbiology of field grown sugar beet. FEMS Microbiol. Ecol. 16: 205–212.

    Article  CAS  Google Scholar 

  16. Rainey, P.B., Bailey, M.J. and Thompson, I.P. 1994 Phenotypic and geno-typic diversity of fluorescent pseudomonads isolated from field grown sugar beet. Microbiology 140: 2315–2331.

    Article  CAS  PubMed  Google Scholar 

  17. Bailey, M.J., Lilley, A.K., Thompson, I.P., Rainey, P.B. and Ellis, R.J. 1995. Site directed chromosomal marking of a fluorescent pseudomonad isolated from the phytosphere of sugar beet; stability and potential for marker gene transfer. Molec. Ecol. In press.

  18. Drahos, D.J., Hemming, B.C. and McPherson, S. 1986. Tracking recombinant organisms in the environment: beta-galactosidase as a selectable, non-antibiotic marker for fluorescent pseudomonads. Bio/Technology 4: 439–443.

    CAS  Google Scholar 

  19. Barry, G.F. 1988. A broad host range shuttle system for gene insertion into the chromosome of Gram-negative bacteria. Gene 71: 75–84.

    Article  CAS  PubMed  Google Scholar 

  20. Rainey, P.B. and Bailey, M.J. 1995. Physical and genetic map of the Pseudomonas fluorescens SBW25 chromosome. Molec. Microbiol. In press.

  21. De Leij, F.A.A.M., Bailey, M.J., Lynch, J.M. and Whipps, J.M. 1993. A simple most probable number technique for the sensitive recovery of a genetically engineered Pseudomonas aureofaciens from soil. Lett. Appl. Microbiol. 16: 307–310.

    Article  Google Scholar 

  22. Bramwell, P.A., Barallon, R.V., Rogers, H.J. and Bailey, M.J. 1995. Extraction and PCR amplification of DNA from the rhizoplane, p. 36–55. In: Molecular Microbial Ecology Manual. Akkermans, A. D. L., Van Elsas, J. D. and De Bruijn, F. J. (Eds.). Kluwer Academic Publishers.

    Google Scholar 

  23. Thompson, I.P., Ellis, R.J. and Bailey, M.J. 1995. Autecology of a genetically modified fluorescent pseudomonad on sugar beet. FEMS Microbiol. Ecol. 17: 1–14.

    Article  CAS  Google Scholar 

  24. Ellis, R.J., Thompson, I.P. and Bailey, M.J. 1995. Metabolic profiling as a means of characterising plant-associated microbial communities. FEMS Microbiol. Ecol. 16: 9–18.

    Article  CAS  Google Scholar 

  25. De Leij, F.A.A.M., Sutton, E.J., Whipps, J.M. and Lynch, J.M. 1994. Spread and survival of a genetically modified Pseudomonas aureofaciens in the phytosphere of wheat and in soil. Appl. Soil. Ecol. 1: 207–218.

    Article  Google Scholar 

  26. Anon. 1995. Risk assessment and genetically modified microorganisms: Final phylloplane report. Research report no. 5, Department of the Environment, London.

  27. Anon. 1995. Risk assessment and genetically modified microorganisms: Final rhizosphere report. Research report no. 6, Department of the Environment, London.

  28. Anon. 1995. Risk assessment and genetically modified microorganisms: Final release report. Research report no. 7, Department of the Environment, London.

  29. De Leij, F.A.A.M., Sutton, E.J., Whipps, J.M., Fenlon, J.S. and Lynch, J.M. 1995. Field release of a genetically modified Pseudomonas fluorescens on wheat: establishment, survival and dissemination. Bio/Technology 13: xxx–xxx.

    Google Scholar 

  30. O'Sullivan, D.B. and O'Gara, F. 1992. Traits of fluorescent Pseudomonas spp. involved in suppression of plant root pathogens. Microbiol. Rev. 56: 662–676.

    CAS  PubMed  PubMed Central  Google Scholar 

  31. Lenski, R.E. 1991. Quantifying fitness and gene stability in micro-organisms, p. 173–192. In: Assessing Ecological Risks of Biotechnology. Grinzburg, L. R. (Ed.). Butterworth-Heinemann, Boston.

    Chapter  Google Scholar 

  32. Hirano, S.S., Nordheim, E.V., Arny, D.C. and Upper, C.D. 1982. Lognormal distribution of epiphytic bacterial populations on leaf surfaces. Appl. Environ. Microbiol. 44: 695–700.

    CAS  PubMed  PubMed Central  Google Scholar 

  33. Lilley, A.K., Fry, J.C., Day, M.J. and Bailey, M.J. 1994. In situ transfer of an exogenously isolated plasmid between indigenous donor and recipient Pseudomonad spp in sugar beet rhizosphere. Microbiology 140: 27–33.

    Article  CAS  Google Scholar 

  34. Piper, K.R., Beck von Bodman, S. and Farrand, S.K. 1993. Conjugation of Agrobacterium tumefaciens regulates Ti plasmid transfer by autoinduction. Nature 362: 448–450.

    Article  CAS  PubMed  Google Scholar 

  35. Kidambi, S.P., Ripp, S. and Miller, R.V. 1994. Evidence for phage-mediated gene transfer among Pseudomonas aeruginosa strains on the phylloplane. Appl. Environ. Microbiol. 60: 496–500.

    CAS  PubMed  PubMed Central  Google Scholar 

  36. Wilson, M. and Lindow, S.E. 1994. Inoculum density dependent mortality and colonisation of the phyllosphere by Pseudomonas syringae. Appl. Environ. Microbiol. 60: 2232–2237.

    CAS  PubMed  PubMed Central  Google Scholar 

  37. Wilson, M. and Lindow, S.E. 1993. Effect of phenotypic plasticity on epiphytic survival and colonisation by Pseudomonas syringae. Appl. Environ. Micro. 59: 410–416.

    CAS  Google Scholar 

  38. Rainey, P.B., Moxon, E.R. and Thompson, I.P. 1994. Intraclonal polymorphism in bacteria, p. 263–293. In: Advances in Microbial Ecology. Jones, J. G. (Ed.). Plenum Press, London

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Natural Environment Research Council, Institute of Virology and Environmental Microbiology, Mansfield Road, Oxford, OX1 3SR, UK

    Ian P. Thompson, Andrew K. Lilley, Richard J. Ellis, Penny A. Bramwell & Mark J. Bailey

Authors
  1. Ian P. Thompson
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Andrew K. Lilley
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Richard J. Ellis
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Penny A. Bramwell
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Mark J. Bailey
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Mark J. Bailey.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Thompson, I., Lilley, A., Ellis, R. et al. Survival, Colonization and Dispersal of Genetically Modified Pseudomonas fluorescens SBW25 in the Phytosphere of Field Grown Sugar Beet. Nat Biotechnol 13, 1493–1497 (1995). https://doi.org/10.1038/nbt1295-1493

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt1295-1493

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing