Skip to main content

Advertisement

SpringerLink
Log in

A Highly Selectable and Highly Transferable Ti Plasmid to Study Conjugal Host Range and Ti Plasmid Dissemination in Complex Ecosystems

  • Published:
Microbial Ecology Aims and scope Submit manuscript

Abstract

A conjugal donor system, ST2, was constructed to study the conjugal dissemination of a Ti plasmid to wild-type recipient bacteria in vitro and in situ. The system consisted of a polyauxotrophic derivative of C58 harboring a hyperconjugative and highly selectable Ti plasmid, pSTiEGK, which was constructed by inserting a multiple antibiotic resistance cassette in the traM-mcpA region of pTiC58ΔaccR. ST2 transfers pSTiEGK constitutively at frequencies up to 10−1 to plasmidless Agrobacterium recipients. The host range of pSTiEGK includes all the known genomic species of Agrobacterium, indigenous soil agrobacteria and some Rhizobium and Phyllobacterium spp. All transconjugants became pathogenic upon acquisition of the Ti plasmid and were also able to transfer pSTiEGK by conjugation. This host range was indistinguishable from that of its wild-type parent pTiC58, and therefore pSTiEGK constitute a valid proxy to study the dissemination of Ti plasmids directly in the environment. Transconjugants can be selected on a combination of four antibiotics, which efficiently prevents the growth of the indigenous microbiota present in complex environments. The transfer of pSTiEGK to members of the genus Agrobacterium was affected primarily by the plasmid content of the recipient strain (103- to 105-fold reduction), e.g., the presence of incompatible plasmids. As a consequence, a species should be considered permissive to Ti transfer whenever one permissive isolate is found.

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

Figure 1

Similar content being viewed by others

References

  1. S Beck von Bodman GT Hayman SK Farrand (1992) ArticleTitleOpine catabolism and conjugal transfer of the nopaline Ti plasmid pTiC58 are coordinately regulated by a single repressor. Proc Natl Acad Sci USA 89 643–647 Occurrence Handle1:STN:280:By2C3MrntFM%3D Occurrence Handle1731335

    CAS  PubMed  Google Scholar 

  2. H Bouzar JB Jones (2001) ArticleTitle Agrobacterium larrymoorei sp. nov., a pathogen isolated from aerial tumours of Ficus benjamina. Int J Syst Evol Microbiol 51 1023–1026 Occurrence Handle1:CAS:528:DC%2BD3MXks1Khsrg%3D Occurrence Handle11411669

    CAS  PubMed  Google Scholar 

  3. DM Cook PL Li F Ruchaud S Padden SK Farrand (1997) ArticleTitleTi plasmid conjugation is independent of vir: Reconstitution of the tra functions from pTiC58 as a binary system. J Bacteriol 179 1291–1297 Occurrence Handle1:CAS:528:DyaK2sXhtFOrtL4%3D Occurrence Handle9023214

    CAS  PubMed  Google Scholar 

  4. Y Dessaux A Petit J Tempé (1992) Opines in Agrobacterium biology. DPS Verma (Eds) Molecular Signals in Plant–Microbe Communications CRC Press Boca Raton, FL 109–136

    Google Scholar 

  5. C Fuqua M Burbea SW Winans (1995) ArticleTitleActivity of the Agrobacterium Ti plasmid conjugal transfer regulator traR is inhibited by the product of the traM gene. J Bacteriol 177 1367–1373 Occurrence Handle1:CAS:528:DyaK2MXktVSnurY%3D Occurrence Handle7868612

    CAS  PubMed  Google Scholar 

  6. C Genetello N Van Larebeke M Holsters A De Picker M Van Montagu J Schell (1977) ArticleTitleTi plasmids of Agrobacterium tumefaciem as conjugative plasmids. Nature 265 561–563 Occurrence Handle1:STN:280:CSiC3c%2FktlY%3D Occurrence Handle834309

    CAS  PubMed  Google Scholar 

  7. GL Grundmann A Dechesne F Bartoli JP Flandrois JL Chasse R Kizungu (2001) ArticleTitleSpatial modeling of nitrifier microhabitats in soil. Soil Sci Soc Am J 65 1709–1716 Occurrence Handle1:CAS:528:DC%2BD38Xht1SmurY%3D

    CAS  Google Scholar 

  8. M Holsters B Silva F van Vliet JP Hernalsteens C Genetello M van Montagu J Schell (1978) ArticleTitle In vivo transfer of the Ti-plasmid of Agrobacterium tumefaciens to Escherichia coli. Mol Gen Genet 163 335–338 Occurrence Handle1:STN:280:CSeB2cnmvFQ%3D Occurrence Handle150536

    CAS  PubMed  Google Scholar 

  9. Hooykaas, PJJ (1979) The role of plasmid determinated functions in the interaction of Rhizobiaceae with plant cells. A genetic approach. PhD thesis, University of Leiden, Leiden, The Netherlands

  10. PJ Hooykaas AGM Beijersbergen (1994) ArticleTitleThe virulence system of Agrobacterium tumefaciens. Ann Rev 32 157–179 Occurrence Handle10.1146/annurev.py.32.090194.001105 Occurrence Handle1:CAS:528:DyaK2cXmtVSntbs%3D

    Article  CAS  Google Scholar 

  11. PJJ Hooykaas H den Dulk-Ras G Ooms RA Schilperoort (1980) ArticleTitleInteractions between octopine and nopaline plasmids in Agrobacterium tumefaciens. J Bacteriol 143 1295–1306 Occurrence Handle1:CAS:528:DyaL3cXmtVSru78%3D Occurrence Handle7410319

    CAS  PubMed  Google Scholar 

  12. PJJ Hooykaas H den Dulk-Ras RA Schilperoort (1980) ArticleTitleMolecular mechanism of Ti-plasmid mobilization by R-plasmid isolation of Ti-plasmid with transposon insertions in Agrobacterium tumefaciens. Plasmid 4 65–74

    Google Scholar 

  13. PJJ Hooykaas PM Klapwijk MP Nuti RA Schilperoort A Rorsch (1977) ArticleTitleTransfer of the Agrobacterium tumefaciens Ti-plasmid to avirulent agrobacteria and to Rhizobium explanta. J Gen Microbiol 98 477–484

    Google Scholar 

  14. I Hwang DM Cook SK Farrand (1995) ArticleTitleA new regulatory element modulates homoserine lactone-mediated autoinduction of Ti plasmid conjugal transfer. J Bacteriol 177 449–458 Occurrence Handle1:CAS:528:DyaK2MXjtFOqsb4%3D Occurrence Handle7814335

    CAS  PubMed  Google Scholar 

  15. A Kerr (1969) ArticleTitleTransfer of virulence between isolates of Agrobacterium. Nature 223 1175–1176

    Google Scholar 

  16. A Kerr P Manigault J Tempé (1977) ArticleTitleTransfer of virulence in vivo and in vitro in Agrobacterium. Nature 265 560–561 Occurrence Handle1:STN:280:CSiC3c%2FktlU%3D Occurrence Handle834308

    CAS  PubMed  Google Scholar 

  17. PM Klapwijk RA Schilperoort (1979) ArticleTitleNegative control of octopine degradation and transfer genes of octopine Ti plasmids in Agrobacterium tumefaciens. J Bacteriol 139 424–431 Occurrence Handle1:CAS:528:DyaE1MXltlKqu7c%3D Occurrence Handle457610

    CAS  PubMed  Google Scholar 

  18. T Komari W Halperin EW Nester (1986) ArticleTitlePhysical and functional map of supervirulent Agrobacterium tumefaciens tumor-inducing plasmid pTiBo542. J Bacteriol 166 88–94 Occurrence Handle1:CAS:528:DyaL28XhvVSkurc%3D Occurrence Handle3957875

    CAS  PubMed  Google Scholar 

  19. Z Krimi A Petit C Mougel Y Dessaux X Nesme (2002) ArticleTitleSeasonal fluctuations and long-term persistence of pathogenic populations of Agrobacterium spp. in soils. Appl Environ Microbiol 68 3358–3365 Occurrence Handle10.1128/AEM.68.7.3358-3365.2002 Occurrence Handle1:CAS:528:DC%2BD38Xlt1Snt7o%3D Occurrence Handle12089015

    Article  CAS  PubMed  Google Scholar 

  20. BL Kittell DR Helinski (1993) Plasmid incompatibility and replication control. DB Clewell (Eds) Bacterial Conjugation Plenum Press New York 223–242

    Google Scholar 

  21. PL Li DM Everhart SK Farrand (1998) ArticleTitleGenetic and sequence analysis of the pTiC58 trb locus, encoding a mating-pair formation system related to members of the type IV secretion family. J Bacteriol 180 6164–6172 Occurrence Handle1:CAS:528:DyaK1cXnslyisrs%3D Occurrence Handle9829924

    CAS  PubMed  Google Scholar 

  22. M Mersereau GJ Pazour A Das (1990) ArticleTitleEfficient transformation of Agrobacterium tumefaciens by electroporation. Gene 90 149–151 Occurrence Handle10.1016/0378-1119(90)90452-W Occurrence Handle1:CAS:528:DyaK3cXlt1OlsLo%3D Occurrence Handle2165971

    Article  CAS  PubMed  Google Scholar 

  23. HN Miller JW Miller JL Crane (1975) ArticleTitleRelative susceptibility of Chrysanthemum morifolium cultivars to Agrobacterium tumefaciens. Plant Dis Rep 59 576–581

    Google Scholar 

  24. C Mougel B Cournoyer X Nesme (2001) ArticleTitleNovel tellurite-amended media and specific chromosomal and Ti plasmid probes for direct analysis of soil populations of Agrobacterium biovar 1 and 2. Appl Environ Microbiol 67 65–74 Occurrence Handle10.1128/AEM.67.1.65-74.2001 Occurrence Handle1:CAS:528:DC%2BD3MXjtVWhsw%3D%3D Occurrence Handle11133429

    Article  CAS  PubMed  Google Scholar 

  25. C Mougel J Thioulouse G Perrière X Nesme (2002) ArticleTitleA mathematical method for determining genome divergence and species delineation using AFLP. Int J Syst Evol Microbiol 52 573–586 Occurrence Handle1:CAS:528:DC%2BD38XivV2rurY%3D Occurrence Handle11931171

    CAS  PubMed  Google Scholar 

  26. DA Morrison MC Trombe MK Hayden GA Waszak JD Chen (1984) ArticleTitleIsolation of transformation-deficient Streptococcus pneumoniae mutants defective in control of competence using insertion–duplication mutagenesis with the erythromycin resistance determinant of pAMb1. J Bacteriol 159 870–876 Occurrence Handle1:CAS:528:DyaL2cXmt1elt7g%3D Occurrence Handle6090396

    CAS  PubMed  Google Scholar 

  27. J Murillo H Shen D Gerhold A Sharma DA Cooksey N Keen (1994) ArticleTitleCharacterization of pPT23B the plasmid involved in syringolide production by Pseudomonas syringae pv. tomato PT23. Plasmid 34 275–287 Occurrence Handle10.1006/plas.1994.1029

    Article  Google Scholar 

  28. X Nesme C Picard P Simonet (1995) Specific DNA sequences for detection of soil bacteria. JT Trevor JD van Elsas (Eds) Nucleic Acids in the Environment Springer-Verlag Berlin 111–139

    Google Scholar 

  29. P Oger KS Kim RL Sackett KR Piper SK Farrand (1998) ArticleTitleOctopine-type Ti plasmids code for a mannopine-inducible dominant-negative allele of traR, the quorum-sensing activator that regulates Ti plasmid conjugal transfer. Mol Microbiol 27 277–288 Occurrence Handle10.1046/j.1365-2958.1998.00671.x Occurrence Handle1:CAS:528:DyaK1cXotF2ktg%3D%3D Occurrence Handle9484884

    Article  CAS  PubMed  Google Scholar 

  30. K Ophel A Kerr (1990) ArticleTitle Agrobacterium vitis, sp. nov. for strains of Agrobacterium biovar 3 from grapevine. Int J Syst Bacteriol 40 236–241 Occurrence Handle1:CAS:528:DyaK3cXls1yrurg%3D

    CAS  Google Scholar 

  31. A Petit C David GA Dahl JG Ellis P Guyon F Casse-Delbart J Tempé (1983) ArticleTitleFurther extension of the opine concept: plasmids in Agrobacterium rhizogenes cooperate for opine degradation. Mol Gen Genet 190 204–214 Occurrence Handle1:CAS:528:DyaL3sXltFKqs7o%3D

    CAS  Google Scholar 

  32. A Petit J Tempé (1978) ArticleTitleIsolation of Agrobacterium Ti-plasmid regulatory mutants. Mol Gen Genet 167 147–155 Occurrence Handle1:CAS:528:DyaE1MXpvF2isw%3D%3D

    CAS  Google Scholar 

  33. A Petit J Tempé M Holsters J van Montagu J Schell (1978) ArticleTitleSubstrate induction of conjugative activity of Agrobacterium tumefaciens Ti plasmids. Nature 271 570–572 Occurrence Handle1:CAS:528:DyaE1cXhs1Kmt74%3D

    CAS  Google Scholar 

  34. C Ponsonnet X Nesme (1994) ArticleTitleIdentification of Agrobacterium strains by PCR-RFLP analysis of pTi and chromosomal regions. Arch Microbiol 161 300–309 Occurrence Handle10.1007/s002030050058 Occurrence Handle1:CAS:528:DyaK2cXkt1yjsbo%3D Occurrence Handle7911654

    Article  CAS  PubMed  Google Scholar 

  35. MY Popoff K Kersters M Kiredjian I Miras C Coynault (1984) ArticleTitlePosition taxonomique de souches d’Agrobacterium d’origine hospitalière. Ann Microbiol, Inst Pasteur 135 427–442

    Google Scholar 

  36. C Rosenberg T Huguet (1984) ArticleTitleThe pAtC58 plasmid of Agrobacterium tumefaciens is not essential for tumor induction. Mol Gen Genet 196 533–536 Occurrence Handle1:CAS:528:DyaL2cXmt1elsL0%3D

    CAS  Google Scholar 

  37. J Sambrook EF Fritsch T Maniatis (1989) Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press Cold Spring Harbor, NY

    Google Scholar 

  38. E Stackebrandt W Frederiksen GM Garrity P Grimont P Kämpfer MCJ Maiden X Nesme R Rosselló-Mora J Swings HG Trüper L Vauterin AC Ward WB Whitman (2002) ArticleTitleReport of the Ad Hoc Committee for the Re-Evaluation of the Species Definition in Bacteriology. Int J Syst Evol Microbiol 52 1043–1047

    Google Scholar 

  39. J Tempé P Guyon DA Tepfer A Petit (1979) The role of opines in the ecology of the Ti plasmids of Agrobacterium. KN Timmis A Pühler (Eds) Plasmids of Medical, Environmental and Commercial Importance Elsevier/North Holland Biomedical Press, Amsterdam 353–365

    Google Scholar 

  40. N Van Larebeke G Engler M Holsters S van den Elsacker I Zaenen RA Schilperoort J Schell (1974) ArticleTitleLarge plasmid in Agrobacterium tumefaciens essential for crown gall-inducing ability. Nature 252 169–170 Occurrence Handle1:STN:280:CSqD383hslE%3D Occurrence Handle4419109

    CAS  PubMed  Google Scholar 

  41. RJM Van Veen H den Dulk-Ras T Bisseling RA Schilperoot PJJ Hooykaas (1988) ArticleTitleCrown gall tumor and root nodule formation by the bacterium Phyllobacterium myrsinacearum after the introduction of an Agrobacterium Ti plasmid or a Rhizobium Sym plasmid. Mol Plant–Microbe Interact 6 231–234

    Google Scholar 

  42. V Vaudequin-Dransart A Petit C Poncet C Ponsonnet X Nesme JB Jones H Bouzar WS Chilton Y Dessaux (1995) ArticleTitleNovel Ti plasmids in Agrobacterium strains isolated from Ficus and Chrysanthemum tumors and their opine-related molecules. Mol Plant–Microbe Interact 8 311–321 Occurrence Handle1:CAS:528:DyaK2MXls1Oku70%3D

    CAS  Google Scholar 

  43. J Vogel P Normand J Thioulouse X Nesme G Grundmann (2003) ArticleTitleRelationship between spatial and genetic distance in Agrobacterium spp. in 1 cubic centimeter of soil. Appl Environ Microbiol 69 1482–1487 Occurrence Handle10.1128/AEM.69.3.1482-1487.2003 Occurrence Handle1:CAS:528:DC%2BD3sXitlClsrk%3D Occurrence Handle12620832

    Article  CAS  PubMed  Google Scholar 

  44. R Wheatcroft DG McRae RW Miller (1990) ArticleTitleChanges in the Rhizobium meliloti genome and the ability to detect supercoiled plasmids during bacteroid development. Mol Plant–Microbe Interact 3 9–17 Occurrence Handle1:CAS:528:DyaK38XhtVSmu7k%3D

    CAS  Google Scholar 

  45. FF White EW Nester (1980) ArticleTitleHairy root: Plasmid encodes virulence traits in Agrobacterium rhizogenes. J Bacteriol 141 1134–1141 Occurrence Handle1:CAS:528:DyaL3cXhvFChu7k%3D Occurrence Handle6245060

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

The authors thank P. Hooykaas and Y. Dessaux for many of the strains used in this study. Thanks are also adressed to Marie-André Poirier for her skillful technical assistance. This work and K.G. were supported by grant ERB1C18CT970198 from the European Union program INCO-DC to X.N. S.T.-C. received a fellowship from French Research Ministry (MENSR). Portion of this work by P.O. and S.T.-C. were supported by C-FAR grant 99I-059-4 from the State of Illinois and by grant R01-GM52465 from the NIH to S.K.F.

Author information

Authors and Affiliations

  1. Ecologie Microbienne, Université Claude Bernard—Lyon 1, USC CNRS 5557, LRC INRA 1193, 16 rue Raphaël Dubois, F-69622 Villeurbanne cedex, France

    S. Teyssier-Cuvelle, C. Mougel, K. Groud & X. Nesme

  2. Department of Crop Sciences, University of Illinois at Urbana–Champaign, Urbana, IL 61801, USA

    P. Oger & S.K. Farrand

  3. Department of Microbiology, University of Illinois at Urbana–Champaign, Urbana, IL 61801, USA

    S.K. Farrand

Authors
  1. S. Teyssier-Cuvelle
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. P. Oger
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. C. Mougel
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. K. Groud
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. S.K. Farrand
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. X. Nesme
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to X. Nesme.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Teyssier-Cuvelle, S., Oger, P., Mougel, C. et al. A Highly Selectable and Highly Transferable Ti Plasmid to Study Conjugal Host Range and Ti Plasmid Dissemination in Complex Ecosystems . Microb Ecol 48, 10–18 (2004). https://doi.org/10.1007/s00248-003-2023-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00248-003-2023-6

Keywords

Search

Navigation