Skip to main content
Log in

Ti plasmid containing Rhizobium meliloti are non-tumorigenic on plants, despite proper virulence gene induction and T-strand formation

  • Original Papers
  • Published:
Archives of Microbiology Aims and scope Submit manuscript

Abstract

We examined the expression of the vir genes of the Agrobacterium tumefaciens Ti plasmid in Rhizobium meliloti, which remains non-tumorigenic on plants after introduction of a Ti- or Ri-plasmid. Both the levels of virulence (vir) gene expression, induced by the plant phenolic compound acetosyringone, and of subsequent T-strand formation were comparable to what is observed in Agrobacterium. In contrast to the situation in Agrobacterium, though, vir induction in R. meliloti did not require a low pH (5.3) of the induction medium and the optimum temperature for induction in R. meliloti was significantly lower than in Agrobacterium. At 37°C no induction of the vir genes was found both in Agrobacterium and R. meliloti. We postulate that the lack of tumorigenicity of Ti carrying R. meliloti strains is due either to a lack of proper attachment of the bacteria to plant cells, or to an improper assembly of a virB-determined essential structure in the cell wall of R. meliloti.

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

  • Alt-MoerbeJ, NeddermannP, LintigJvon, WeilerEW, SchröderJ (1988) Temperature sensitive step in Ti plasmid vir-region induction and correlation with cytokinin secretion by Agrobacterium. Mol Gen Genet 213: 1–8

    Google Scholar 

  • BeringerJE, BeynonJL (1978) Transfer of the drug-resistance transposon Tn5 to Rhizobium. Nature 276: 633–634

    Google Scholar 

  • Caetano AnollesG, FavelukesG (1986) Host-symbiont specificity expressed during early adsorption of Rhizobium meliloti to the root surface of alfalfa. Appl Environm Microbiol 52: 377–383

    Google Scholar 

  • DouglasC, HarperinW, NesterEW (1982) Agrobacterium tumefaciens mutants affected in attachment to plant cells. J Bacteriol 152: 1265–1275

    Google Scholar 

  • DraperJ, MackensieIA, DaveyMR, FreemanJP (1983) Attachment of Agrobacterium tumefaciens to mechanically isolated Asparagus cells. Plant Science Lett 29: 227–236

    Google Scholar 

  • GietlC, Koukolikova-NicolaZ, HohnB (1987) Mobilization of T-DNA from Agrobacterium to plant cells involves a protein that binds single-stranded DNA. Proc Natl Acad Sci USA 84: 9006–9010

    Google Scholar 

  • HooykaasPJJ, SchilperoortRA (1984) The molecular genetics of crown-gall tumorigenesis. In: ScandaliosHG (ed) Molecular genetics of plants (Advances in genetics vol 22). Academic Press, Orlando, USA, pp 209–283

    Google Scholar 

  • HooykaasPJJ, KlapwijkPM, NutiMP, SchilperoortRA, RörschA (1977) Transfer of the Agrobacterium tumefaciens Ti plasmid to avirulent agrobacteria and to Rhizobium ex planta. J Gen Microbiol 98: 477–484

    Google Scholar 

  • HooykaasPJJ, RoobolC, SchilperoortRA (1979) Regulation of the transfer of Ti plasmids of Agrobacterium tumefaciens. J Gen Microbiol 110: 99–109

    Google Scholar 

  • HooykaasPJJ, SnijdewindtFGM, SchilperoortRA (1982) Identification of the Sym plasmid of Rhizobium leguminosarum strain 1001 and its transfer to and expression in other rhizobia and Agrobacterium tumefaciens. Plasmid 8: 73–82

    Google Scholar 

  • KadoCI, LiuST (1981) Rapid procedure for detection and isolation of large and small plasmids. J Bacteriol 145: 1365–1373

    Google Scholar 

  • Koukolikova-NicolaZ, AlbrightL, HohnB (1987) The mechanism of T-DNA transfer from Agrobacterium tumefaciens to plant cells. In: HohnT, SchellJ (eds) Plant DNA infectious agents. Springer, Wien, pp 109–148

    Google Scholar 

  • KrensFA, MolendijkL, WullemsGJ, SchilperoortRA (1985) The role of bacterial attachment in the transformation of cell-wall-regenerating tobacco protoplasts by Agrobacterium tumefaciens. Planta 166: 300–308

    Google Scholar 

  • LafreniereC, BordeleauLM, AmargerN, AntainH (1984) Effect of heterologous bacteria and combined nitrogen on the adsorption of Rhizobium meliloti to lucerne seedling roots. Plant Soil 82: 223–229

    Google Scholar 

  • LerouxB, YanofskyMF, WinansSC, WardJE, ZieglerSF, NesterEW (1987) Characterization of the virA locus of Agrobacterium tumefaciens: a transcriptional regulator and host-range determinant. EMBO J 6: 849–856

    Google Scholar 

  • MatthysseAG (1986) Initial interactions of Agrobacterium tumefaciens with plant host cells. CRC Crit Rev in Microbiol 13: 281–307

    Google Scholar 

  • MatthysseAG (1987) Characterization of non-attaching mutants of Agrobacterium tumefaciens. J Bacteriol 169: 313–323

    Google Scholar 

  • Melchers LS, Hooykaas PJJ (1987) Virulence of Agrobacterium. In: Oxford surveys of plant molecular and cell biology, vol 4, pp 167–220

  • MelchersLS, ThompsonDV, IdlerKB, SchilperoortRA, HooykaasPJJ (1986) Nucleotide sequence of the virulence gene virG of the Agrobacterium tumefaciens octopine Ti plasmid: significant homology between virG and the regulatory genes ompR, phoB and dye of E. coli. Nucl Acids Res 14: 9933–9942

    Google Scholar 

  • MelchersLS, ThompsonDV, IdlerKB, NeuteboomSTC, deMaagdRA, SchilperoortRA, HooykaasPJJ (1987) Molecular characterization of the virulence gene virA of the Agrobacterium tumefaciens octopine Ti plasmid. Plant Mol Biol 11: 227–237

    Google Scholar 

  • Melchers LS, Regenburg-Tuïnk AJG, Schilperoort RA, Hooykaas PJJ (1989) Specificity of signal molecules in the activation of Agrobacterium virulence gene expression. Mol Microbiol (in press)

  • MemelinkJ, dePaterBS, HogeJHC, SchilperoortRA (1987) T-DNA hormone biosynthetic genes: phytohormones and gene expression in plants. Devel Genet 8: 321–337

    Google Scholar 

  • MillerJH (1972) In: Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y., pp 352–355

    Google Scholar 

  • OomsG, HooykaasPJJ, MoolenaarG, SchilperoortRA (1981) Crown gall plant tumors of abnormal morphology, induced by Agrobacterium tumefaciens carrying mutated octopine Ti plasmids; analysis of T-DNA functions. Gene 14: 33–50

    Google Scholar 

  • VanSchieBJ, DijkenvanJP, KuenenJG (1984) Non-coordinated synthesis of glucose dehydrogenase and its prostethic group PQQ in Acinetobacter and Pseudomonas species. FEMS Microbiol Lett 24: 133–138

    Google Scholar 

  • Smit G (1988) Adhesins from Rhizobiaceae and their role in plant-bacterium interactions. PhD Thesis, Leiden University

  • StachelSE, MessensE, VanMontaguM, ZambryskiP (1985) Identification of the signal molecules produced by wounded plant cells that activate T-DNA transfer in Agrobacterium tumefaciens. Nature 318: 624–629

    Google Scholar 

  • StachelSE, TimmermanB, ZambryskiP (1986) Generation of single-stranded T-DNA molecules during the initial stages of T-DNA transfer from Agrobacterium tumefaciens to plant cells. Nature 322: 706–712

    Google Scholar 

  • StachelSE, TimmermanB, ZambryskiP (1987) Activation of Agrobacterium tumefaciens vir gene expression generates multiple single stranded T-strand molecules from the pTiA6 T-region: requirement for 5′ virD gene products. EMBO J 6: 857–863

    Google Scholar 

  • ThomashowMF, KarlinseyJE, MarksJR, HurlbertRE (1987) Identification of a new virulence locus in Agrobacterium tumefaciens that affects polysaccharide composition and plant cell attachment. J Bacteriol 169: 3209–3216

    Google Scholar 

  • ThompsonDV, MelchersLS, IdlerKB, SchilperpoortRA, hooykaasPJJ (1988) Analysis of the complete nucleotide sequence of the Agrobacterium tumefaciens virB operon. Nucl Acids Res 16: 4621–4636

    Google Scholar 

  • VanHaarenMJJ, SedeeNJA, SchilperoortRA, HooykaasPJJ (1987) Overdrive is a T-region transfer enhancer which stimulates T-strand production in Agrobacterium tumefaciens. Nucl Acids Res 15: 8983–8997

    Google Scholar 

  • VeenRJMvan, HooykaasPJJ, SchilperoortRA (1988a) Mechanisms of tumorigenesis by Agrobacterium tumefaciens. In: KeenNT, WallingU (eds) Physiology and biochemistry of plant microbial interactions. Waverly Press Inc, Baltimore, USA, pp 19–30

    Google Scholar 

  • VeenRJMvan, Dulk-RasHden, BisselingT, SchilperoortRA, HooykaasPJJ (1988b) Crown gall tumor and root nodule formation by the bacterium Phyllobacterium myrsinacearum after the introduction of an Agrobacterium Ti plasmid or a Rhizobium Sym plasmid. Molec. Plant-Microbe Interactions 1: 231–234

    Google Scholar 

  • WardJE, AkiyoshiDE, RegiersD, DattaA, GordonM, NesterEW (1988) Characterization of the virB operon from an Agrobacterium tumefaciens Ti plasmid. J Biol Chem 263: 5804–5814

    Google Scholar 

  • WinansSC, EbertPR, StachelSE, GordonMP, NesterEW (1986) A gene essential for Agrobacterium virulence is homologous to a family of positive regulatory loci. Proc Natl Acad Sci USA 83: 8278–8282

    Google Scholar 

  • YanofskyMF, PorterSG, YoungC, AlbrightLM, GordonMP, NesterEW (1986) The virD operon of Agrobacterium tumefaciens encodes a site-specific endonuclease. Cell 47: 471–477

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

van Veen, R.J.M., den Dulk-Ras, H., Schilperoort, R.A. et al. Ti plasmid containing Rhizobium meliloti are non-tumorigenic on plants, despite proper virulence gene induction and T-strand formation. Arch. Microbiol. 153, 85–89 (1989). https://doi.org/10.1007/BF00277546

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00277546

Key words

Navigation