Skip to main content
SpringerLink
Log in

Mechanisms of crown gall formation: T-DNA transfer fromAgrobacterium tumefaciens to plant cells

  • Invited Article
  • Published:
The botanical magazine = Shokubutsu-gaku-zasshi Aims and scope Submit manuscript

Abstract

Agrobacterium tumefaciens harbouring the Ti plasmid incites crown gall tumor on dicotyledonous species. Upon infection of these plants, T-DNA in the Ti plasmid is transferred by unknown mechanisms to plant cells to be integrated into nuclear DNA. WhenAgrobacterium is incubated with protoplasts or seedlings of dicotyledonous plants, circulation of T-DNA and expression ofvir (virulence) genes on the Ti plasmid are induced. The circularization event is efficiently induced by mesophyll protoplasts of tobacco which are highly competent for transformation by the T-DNA, and is also induced by diffusible phenolic compounds excreted from the protoplasts. The circularization and formation of crown gall both require the expression of thevirD locus, one of the induciblevir genes. These results suggest that the circularization of T-DNA reflects one of steps of the T-DNA transfer during formation of crown gall. In contrast to dicotyledonous plants, monocotyledonous plants are thought to be unresponsive to infection byAgrobacterium. We showed that monocotyledonous plants do not excrete diffusible inducers for the expression ofvir genes, while they contain a novel type of a signal substance(s). This inducer is not detected in the exudates of seedlings of monocotyledonous plants, but is found in the extracts from the seedlings, and also those from the seeds, bran and germ of wheat and oats. This finding suggests that T-DNA processing, and possibly its transfer, should take place whenAgrobacterium invades seedlings and seeds of monocotyledonous plants.

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

Similar content being viewed by others

References

  • Akiyoshi, D.E., O.R. Morris, R. Hinz, B.S. Mischke, T. Kosuge, D.J. Garfinkel, M.P. Gordon andE.W. Nester. 1983. Cytokinin/auxin balance in crown gall tumors is regulated by specific loci in the T-DNA. Proc. Nat. Acad. Sci. USA80: 307–411.

    Article  Google Scholar 

  • Albright, L.M., B. Leroux, M.F. Yanofsky, D. Ma andE.W. Nester. 1987. Processing of the T-DNA ofAgrobacterium tumefaciens generates a single-stranded linear DNA intermediate. J. Bacteriol.169: 1046–1055.

    PubMed  CAS  Google Scholar 

  • Barry, G.F., S.G. Rogers, R.T. Fraley andL. Brand. 1984. Identification of a cloned cytokinin biosynthetic gene. Proc. Nat. Acad. Sci. USA81: 4776–4780.

    Article  PubMed  CAS  Google Scholar 

  • Bevan, M.W. andM.-D. Chilton. 1982. T-DNA of the agrobacterium Ti and Ri plasmids. Ann. Rev. Genet.16: 357–384.

    Article  PubMed  CAS  Google Scholar 

  • Bolton, G.D., D.W. Nester andM.P. Gordon. 1986. Plant phenolic compounds induce expression of theAgrobacterium tumefaciens loci needed for virulence. Science232: 983–985.

    PubMed  CAS  Google Scholar 

  • Braun, A.C. 1958. A physiological basis for autonomous growth of the crown-gall tumor cell. Proc. Nat. Acad. Sci. USA44: 344–349.

    Article  PubMed  CAS  Google Scholar 

  • Chilton, M.-D., M.H. Drummond, D.J. Merlo, D. Sciaky, A.L. Montoya, M.P. Gordon andE.W. Nester. 1977. Stable incorporation of plasmid DNA into higher plant cells: the molecular basis of crown gall tumorigenesis. Cell11: 263–271.

    Article  PubMed  CAS  Google Scholar 

  • Close, T.J., P.M. Rogowsky, C.I. Kado, S.C. Winans, M.F. Yanofsky andE.W. Nester. 1987. Dual control ofAgrobacterium tumefaciens Ti plasmid virulence genes. J. Bacteriol.169: 5113–5118.

    PubMed  CAS  Google Scholar 

  • Douglas, C.J., R.J.R.J. Staneloni, R.A. Rubin andE.W. Nester. 1985. Identification and genetic analysis of anAgrobacterium tumefaciens chromosomal virulence region. J. Bacteriol.161: 850–860.

    PubMed  CAS  Google Scholar 

  • Garfinkel, D.J. andE.W. Nester. 1980.Agrobacterium tumefaciens mutants affected in crown gall tumorigenesis and octopine catabolism. J. Bacteriol.144: 732–743.

    PubMed  CAS  Google Scholar 

  • Graves, A.C.F. andS.L. Goldman. 1986. The transformation ofZea mays seedlings withAgrobacterium tumefaciens. Plant Mol. Biol.7: 43–50.

    Article  CAS  Google Scholar 

  • Grimsly, N., T. Hohn, J.W. Davies andB. Hohn. 1987.Agrobacterium-mediated delivery of infectious maize streak virus into maize plants. Nature325: 177–179.

    Article  Google Scholar 

  • Hazelbauer, G.L. andJ. Adler. 1971. Role of the galactose binding protein in chemotaxis ofEscherichia coli toward galactose. Nature New Bio.230: 101–104.

    CAS  Google Scholar 

  • Hazelbauer, G.L. andS. Harayama. 1983. Sensory transduction in bacterial chemotaxis. Inter. Rev. Cytol.81: 33–70.

    Article  CAS  Google Scholar 

  • Hernalsteens, J.-P., L. Thia-Toong, J. Schell andM. Van Montagu. 1984. AnAgrobacterium-transformed cell culture from the monocotAsparagus officinalis. EMBO J.3: 3039–3041.

    PubMed  CAS  Google Scholar 

  • Herrera-Estrella, A., Z.-M. Chen, M.V. Montagu andK. Wang. 1988. VirD proteins ofAgrobacterium tumefaciens are required for the formation of a covalent DNA-protein complex at the 5′ terminus of T-strand molecules. EMBO J.7: 4055–4062.

    PubMed  CAS  Google Scholar 

  • Hille, J., J. van Kan andR.A. Schilperoort. 1984. Trans-acting virulence function of the octopine Ti plasmid fromAgrobacterium tumefaciens. J. Bacteriol.158: 754–756.

    PubMed  CAS  Google Scholar 

  • Hooykaas, P.J.J., M. Hofker, H. Den Dulk-Ras andR.A. Schilperoot. 1984. A Comparison of Virulence Determinants in an Octopine Ti plasmid, a Nopaline Ti plasmid, and an Ri plasmid by Complementation Analysis ofAgrobacterium tumefaciens Mutants. Plasmid11: 195–205.

    Article  PubMed  CAS  Google Scholar 

  • Hooykaas-van Slogteren, G.M.S., P.J.J. Hooykaas andR.A. Schilperoort. 1984. Expression of Ti plasmid genes in monocotyledonous plants infected withAgrobacterium tumefaciens. Nature311: 763–764.

    Article  CAS  Google Scholar 

  • Ínze, D., A. Follin, M. Van Lijsebettens, C. Simoens, C. Genetello, M. Van Montagu andJ. Schell. 1984. Genetic analysis of the individual T-DNA genes ofAgrobacterium tumefaciens; further evidence that two genes are involved in indole-3-acetic acid synthesis. Mol. Gen. Genet.194: 265–274.

    Article  Google Scholar 

  • Iyer, V.N., H.J. Klee andE.W. Nester. 1982. Units of Genetic Expression in the Virulence region of a Plant Tumor-inducing plasmid ofAgrobacterium tumefaciens. Mol. Gen. Genet.188: 418–424.

    Article  PubMed  CAS  Google Scholar 

  • Jayaswal, R.K., K. Veluthambi, S.B. Gelvin andJ.L. Slightom. 1987. Double-stranded cleavage of T-DNA and generation of single-stranded T-DNA molecules inEscherichia coli by avirD encoded border-specific endonuclease fromAgrobacterium tumefaciens. J. Bacteriol.169: 5035–5045.

    PubMed  CAS  Google Scholar 

  • Joos, H., D. Inze, A. Caplan, M. Sormann, M. Van Montagu andJ. Schell. 1983. Genetic analysis of T-DNA transcripts in nopaline crown galls. Cell32: 1057–1067.

    Article  PubMed  CAS  Google Scholar 

  • Klee, H.J., M.P. Gordon andE.W. Nester. 1982. Complementation analysis ofAgrobacterium tumefaciens Ti plasmid mutations affecting oncogenicity. J. Bacteriol.150: 327–331.

    PubMed  CAS  Google Scholar 

  • —,F.F. White, V.N. Iyer, M.P. Gordon andE.W. Nester. 1983. Mutational analysis of the virulence region of anAgrobacterium tumefaciens Ti plasmid. J. Bacteriol.153: 878–883.

    PubMed  CAS  Google Scholar 

  • Koukolíková-Nicola, Z., R.D. Shillito, B. Horhn, K. Wang, M. Van Montagu andP. Zambryski. 1985. Involvement of circular intermediates in the transfer of T-DNA fromAgrobacterium tumefaciens to plant cells. Nature313: 191–195.

    Article  Google Scholar 

  • Leemans, J., R. Deblaere, L. Wilmizer, H. De Greve, JP. Hernalsteens, M. Van Montagu andJ. Schell. 1982. Genetic identification of functions of TL-DNA transcripts in octopine crown galls. EMBO J.1: 147–152.

    PubMed  CAS  Google Scholar 

  • Leroux, B., M.F. Yanofsky, S.C. Winans, J.E. Ward, S.F. Ziegler andE.W. Nester. 1987. Characterization of thevirA locus ofAgrobacterium tumefaciens: a transcriptional regulator and host range determinant. EMBO J.6: 849–856.

    PubMed  CAS  Google Scholar 

  • Machida, Y., C. Machida, H. Ohtsubo andE. Ohtsubo. 1982. Factors determining frequency of plasmid cointegration mediated by insertion sequence IS1. Proc. Nat. Acad. Sci. USA79: 277–281.

    Article  PubMed  CAS  Google Scholar 

  • —,S. Usami, A. Yamamoto, Y. Niwa andI. Takebe. 1986. Plant-inducible recomdination between the 25 bp border sequences of T-DNA inAgrobacterium tumefaciens. Mol. Gen. Genet.204: 374–382.

    Article  CAS  Google Scholar 

  • Niwa, Y., A. Yamamoto, C. Machida, I. Takebe andY. Machida. 1988. Right-hand border regions of octopine T-DNA are recognized by RNA polymerase ofAgrobacterium as well as by VirD1 and VirD2 proteins. Nucleic Acids Res.16: 7647–7661.

    PubMed  CAS  Google Scholar 

  • Ooms, G., P.M. Klapwijk, J.A. Poulis andR.A. Schilperoot. 1980. Characterization of Tn904 insertion in octopine Ti plasmid mutants ofAgrobacterium tumefaciens. J. Bacteriol.144: 82–91.

    PubMed  CAS  Google Scholar 

  • Peralta, E.G. andL.W. Ream. 1985. T-DNA border sequences required for crown gall tumorigenesis. Proc. Nat. Acad. Sci. USA82: 5112–5116.

    Article  PubMed  CAS  Google Scholar 

  • Porter, S.G., M.F. Yanofsky andE.W. Nester. 1987. Molecular characterization of thevirD operon fromAgrobacterium tumefacies. Nucleic Acids Res.15: 7503–7517.

    PubMed  CAS  Google Scholar 

  • Puvanesarajah, V., F.M. Schell, G. Stacey, C.J. Douglas andE.W. Nester. 1985. Role for 2-linked-β-D-glucan in the virulence ofAgrobacterium tumefaciens. J. Bacteriol.164: 102–106.

    PubMed  CAS  Google Scholar 

  • Ream, L.W., M.P. Gordon andE.W. Nester. 1983. Multiple mutations in the T region of theAgrobacterium tumefaciens tumor-inducing plasmid. Proc. Nat. Acad. USA80: 1660–1664.

    Article  CAS  Google Scholar 

  • Schroder, G., S. Waffenschmidt, E.W. Weiler andJ. Schroder. 1984. The T-region of Ti plasmids codes for an enzyme synthesizing indol-3-acetic acid. Eur. J. Biochem.138: 387–391.

    Article  PubMed  CAS  Google Scholar 

  • Shaw, C.H., M.D. Watson, G.H. Carter andC.H. Shaw. 1984. The right hand copy of the nopaline Ti-plasmid 25 bp repeat is required for tumour formation. Nucleic Acids Res.12: 6031–6041.

    PubMed  CAS  Google Scholar 

  • Simpson, R.B., P.J. O'Hara, W. Kwok, A.L. Montoya, C. Lichtenstein, M.P. Gordon andE.W. Nester. 1982. DNA from the A6S/2 crown gall tumor contains scrambled Ti plasmid sequences near its junctions with plant DNA. Cell29: 1005–1014.

    Article  PubMed  CAS  Google Scholar 

  • Smith, M.M. andR.D. Hartley. 1983. Occurrence and nature of ferulic acid substitution of cell-wall polysaccharides in graminaceous plants. Carbohydrate Res.118: 65–80.

    Article  CAS  Google Scholar 

  • Stachel, S.E., G. An, C. Flores andE.W. Nester. 1985a. A Tn3lacZ transposon for random generation of β-galactosidase gene fusions: application of the analysis of gene expression inAgrobacterium. EMBO J.4: 891–898.

    PubMed  CAS  Google Scholar 

  • —,E. Messens, M. Van Montagu andP.C. Zambryski. 1985b. Identification of the signal molecules produced by wounded plant cells that activate T-DNA transfer inAgrobacterium tumefaciens. Nature318: 624–629

    Article  Google Scholar 

  • — andE.W. Nester. 1986. The genetic and transcriptional organization of thevir region of the A6 Ti plasmid ofAgrobacterium tumefaciens. EMBO J.5: 1445–1454.

    PubMed  CAS  Google Scholar 

  • —— andP.C. Zambryski. 1986a. A Plant cell factor inducesAgrobacterium tumefaciens vir gene expression. Proc. Nat. Acad. Sci. USA83: 379–383.

    Article  PubMed  CAS  Google Scholar 

  • —,B. Timmerman andP.C. Zambryski. 1986b. Generation of single-stranded T-DNA molecules during the initial stages of T-DNA transfer fromAgrobacterium tumefaciens to plant cells. Nature322: 706–712.

    Article  CAS  Google Scholar 

  • Takebe, I. andT. Nagata. 1984. Isolation and culture of protoplasts: Tobacco.In I.K. Vasil, ed., Cell Culture and Somatic Genetics of Plants, pp. 328–339. Academic Press, New York.

    Google Scholar 

  • Thomashow, L.S., S. Reeves andM.F. Thomashow. 1984. Crown gall oncogenesis: Evidence that a T-DNA gene from theAgrobacterium Ti plasmid pTiA6 encodes an enzyme that cetalyzes synthesis of indoleacetic acid. Proc. Nat. Acad. Sci. USA81: 5071–5075.

    Article  PubMed  CAS  Google Scholar 

  • Thomashow, M.F., J. Karlinsey, J.R. Marks andR.E. Hurlbert. 1987. Identification of a new virulence locus inAgrobacterium tumefaciens that affects polysaccharide composition and plant cell attachment. J. Bacteriol.169: 3209–3216.

    PubMed  CAS  Google Scholar 

  • Usami, S., S. Morikawa, I. Takebe andY. Machida. 1987. Absence in monocotyledonous plants of the diffusible plant factors inducing T-DNA circularization andvir gene expression inAgrobacterium. Mol. Gen. Genet.209: 221–226.

    Article  CAS  PubMed  Google Scholar 

  • —,S. Okamoto, S. Takebe andY. Machida. 1988. Factor inducingAgrobacterium tumefaciens vir gene expression is present in monocotyledonous plants. Proc. Nat. Acad. Sci. USA.85: 3748–3752.

    Article  PubMed  CAS  Google Scholar 

  • Veluthambi, K., R.K. Jayaswal andS.B. Gelvin. 1987. The virulence genes A, G and D mediate the double-stranded border cleavage of the T-DNA from theAgrobacterium tumefaciens Ti plasmid. Proc. Nat. Acad. Sci. USA.84: 1881–1885.

    Article  PubMed  CAS  Google Scholar 

  • Wang, K., L. Herrera-Estrella, M. Van Montagu andP. Zambryski. 1984. Right 25 bp terminus sequence of the nopaline T-DNA is essential for and determines direction of DNA transfer fromAgrobacterium to the plant genome. Cell38: 455–462.

    Article  PubMed  CAS  Google Scholar 

  • Ward, E.R. andW. Barnes. 1988. VirD2 protein ofAgrobacterium tumefaciens very tightly linked to the 5′ end of T-DNA strand DNA. Science242: 927–930.

    CAS  Google Scholar 

  • Winans, S.C., P.R. Ebert, S.E. Stachel, M.P. Gordon andE.W. Nester. 1986. A gene essential forAgrobacterium virulence is homologous to a family of positive regulatory loci. Proc. Nat. Acad. Sci. USA.83: 8278–8282.

    Article  PubMed  CAS  Google Scholar 

  • Wullems, G.J., L. Molendijk, G. Ooms andR.A. Schilperoot. 1981. Differential expression of crown gall tumor markers in transformants obtained afterin vitro Agrobacterium tumefaciens-induced transformation of cell wall regenerating protoplasts derived fromNicotiana tabacum. Proc. Nat. Acad. Sci. USA.78: 4344–4348.

    Article  PubMed  CAS  Google Scholar 

  • Yamamoto, A., M. Iwahashi, M.F. Yanofsky, E.W. Nester, I. Takebe andY. Machida. 1987. The promoter proximal region in thevirD locus ofAgrobacterium tumefaciens is necessary for the plant-inducible circularization of T-DNA. Mol. Gen. Genet.206: 174–177.

    Article  PubMed  CAS  Google Scholar 

  • Yanofsky, M.F., S.G. Porter, C. Young, L.M. Albright, M.P. Gordon andE.W. Nester. 1986. ThevirD locus ofAgrobacterium tumefaciens encodes a site-specific endonuclease. Cell47: 471–477.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biology, Faculty of Science, Nagoya University, Chikusa-Ku, 464, Nagoya

    Yasunori Machida, Shigehisa Okamoto, Shogo Matsumoto, Shoji Usami, Akiko Yamamoto, Yasuo Niwa, Soo Doo Jeong, Jun Nagamine, Nobuyoshi Shimoda, Chiyoko Machida & Motoko Iwahashi

Authors
  1. Yasunori Machida
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shigehisa Okamoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Shogo Matsumoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Shoji Usami
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Akiko Yamamoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yasuo Niwa
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Soo Doo Jeong
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Jun Nagamine
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Nobuyoshi Shimoda
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Chiyoko Machida
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Motoko Iwahashi
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Recipient of the Botanical Society Award for Young Scientists, 1987.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Machida, Y., Okamoto, S., Matsumoto, S. et al. Mechanisms of crown gall formation: T-DNA transfer fromAgrobacterium tumefaciens to plant cells. Bot Mag Tokyo 102, 331–350 (1989). https://doi.org/10.1007/BF02488571

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02488571

Key words

Search

Navigation