Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1989 Jul;171(7):3696–3703. doi: 10.1128/jb.171.7.3696-3703.1989

Opines stimulate induction of the vir genes of the Agrobacterium tumefaciens Ti plasmid.

K Veluthambi 1, M Krishnan 1, J H Gould 1, R H Smith 1, S B Gelvin 1
PMCID: PMC210113  PMID: 2738020

Abstract

Upon incubation of Agrobacterium tumefaciens A348 with acetosyringone, the vir genes encoded by the Ti (tumor-inducing) plasmid are induced. The addition of certain opines, including octopine, nopaline, leucinopine, and succinamopine, enhanced this induction 2- to 10-fold. The compounds mannopine, acetopine, arginine, pyruvate, and leucine did not stimulate the induction of the vir genes to such an extent. The enhancement of vir gene induction by opines depended on acetosyringone and the genes virA and virG. Opines stimulated the activity of the vir genes, the double-stranded cleavage of the T (transferred)-DNA at the border repeat sequences, and the production of T-strands by the bacterium. The transformation efficiency of cotton shoot tips was markedly increased by the addition of acetosyringone and nopaline at the time of infection.

Full text

PDF
3696

Images in this article

3700Image
on p.3700

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Albright L. M., Yanofsky M. F., Leroux B., Ma D. Q., Nester E. W. Processing of the T-DNA of Agrobacterium tumefaciens generates border nicks and linear, single-stranded T-DNA. J Bacteriol. 1987 Mar;169(3):1046–1055. doi: 10.1128/jb.169.3.1046-1055.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bevan M. W., Chilton M. D. T-DNA of the Agrobacterium Ti and Ri plasmids. Annu Rev Genet. 1982;16:357–384. doi: 10.1146/annurev.ge.16.120182.002041. [DOI] [PubMed] [Google Scholar]
  3. Bolton G. W., Nester E. W., Gordon M. P. Plant phenolic compounds induce expression of the Agrobacterium tumefaciens loci needed for virulence. Science. 1986 May 23;232(4753):983–985. doi: 10.1126/science.3085219. [DOI] [PubMed] [Google Scholar]
  4. Bomhoff G., Klapwijk P. M., Kester H. C., Schilperoort R. A., Hernalsteens J. P., Schell J. Octopine and nopaline synthesis and breakdown genetically controlled by a plasmid of Agrobacterium tumefaciens. Mol Gen Genet. 1976 May 7;145(2):177–181. doi: 10.1007/BF00269591. [DOI] [PubMed] [Google Scholar]
  5. Chilton W. S., Chilton M. D. Mannityl opine analogs allow isolation of catabolic pathway regulatory mutants. J Bacteriol. 1984 May;158(2):650–658. doi: 10.1128/jb.158.2.650-658.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Christie P. J., Ward J. E., Winans S. C., Nester E. W. The Agrobacterium tumefaciens virE2 gene product is a single-stranded-DNA-binding protein that associates with T-DNA. J Bacteriol. 1988 Jun;170(6):2659–2667. doi: 10.1128/jb.170.6.2659-2667.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Christou P., Platt S. G., Ackerman M. C. Opine synthesis in wild-type plant tissue. Plant Physiol. 1986 Sep;82(1):218–221. doi: 10.1104/pp.82.1.218. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Citovsky V., DE Vos G., Zambryski P. Single-Stranded DNA Binding Protein Encoded by the virE Locus of Agrobacterium tumefaciens. Science. 1988 Apr 22;240(4851):501–504. doi: 10.1126/science.240.4851.501. [DOI] [PubMed] [Google Scholar]
  9. Close T. J., Rogowsky P. M., Kado C. I., Winans S. C., Yanofsky M. F., Nester E. W. Dual control of Agrobacterium tumefaciens Ti plasmid virulence genes. J Bacteriol. 1987 Nov;169(11):5113–5118. doi: 10.1128/jb.169.11.5113-5118.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Close T. J., Tait R. C., Kado C. I. Regulation of Ti plasmid virulence genes by a chromosomal locus of Agrobacterium tumefaciens. J Bacteriol. 1985 Nov;164(2):774–781. doi: 10.1128/jb.164.2.774-781.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Das A. Agrobacterium tumefaciens virE operon encodes a single-stranded DNA-binding protein. Proc Natl Acad Sci U S A. 1988 May;85(9):2909–2913. doi: 10.1073/pnas.85.9.2909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Ditta G., Stanfield S., Corbin D., Helinski D. R. Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7347–7351. doi: 10.1073/pnas.77.12.7347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gietl C., Koukolíková-Nicola Z., Hohn B. Mobilization of T-DNA from Agrobacterium to plant cells involves a protein that binds single-stranded DNA. Proc Natl Acad Sci U S A. 1987 Dec;84(24):9006–9010. doi: 10.1073/pnas.84.24.9006. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jayaswal R. K., Veluthambi K., Gelvin S. B., Slightom J. L. Double-stranded cleavage of T-DNA and generation of single-stranded T-DNA molecules in Escherichia coli by a virD-encoded border-specific endonuclease from Agrobacterium tumefaciens. J Bacteriol. 1987 Nov;169(11):5035–5045. doi: 10.1128/jb.169.11.5035-5045.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jensen R. E., Zdybak W. T., Yasuda K., Chilton W. S. A useful synthesis of nopaline, a crown gall tumor metabolite. Biochem Biophys Res Commun. 1977 Apr 25;75(4):1066–1070. doi: 10.1016/0006-291x(77)91490-5. [DOI] [PubMed] [Google Scholar]
  16. Kerr A., Manigault P., Tempé J. Transfer of virulence in vivo and in vitro in Agrobacterium. Nature. 1977 Feb 10;265(5594):560–561. doi: 10.1038/265560a0. [DOI] [PubMed] [Google Scholar]
  17. Knauf V. C., Nester E. W. Wide host range cloning vectors: a cosmid clone bank of an Agrobacterium Ti plasmid. Plasmid. 1982 Jul;8(1):45–54. doi: 10.1016/0147-619x(82)90040-3. [DOI] [PubMed] [Google Scholar]
  18. Leroux B., Yanofsky M. F., Winans S. C., Ward J. E., Ziegler S. F., Nester E. W. Characterization of the virA locus of Agrobacterium tumefaciens: a transcriptional regulator and host range determinant. EMBO J. 1987 Apr;6(4):849–856. doi: 10.1002/j.1460-2075.1987.tb04830.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lippincott J. A., Lippincott B. B., Chang C. C. Promotion of crown-gall tumor growth by lysopine, octopine, nopaline, and carnosine. Plant Physiol. 1972 Feb;49(2):131–137. doi: 10.1104/pp.49.2.131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lippincott J. A., Lippincott B. B. Lysopine and Octopine Promote Crown-Gall Tumor Growth in vivo. Science. 1970 Oct 9;170(3954):176–177. doi: 10.1126/science.170.3954.176. [DOI] [PubMed] [Google Scholar]
  21. Melchers L. S., Thompson D. V., Idler K. B., Schilperoort R. A., Hooykaas P. J. 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. Nucleic Acids Res. 1986 Dec 22;14(24):9933–9942. doi: 10.1093/nar/14.24.9933. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Montoya A. L., Chilton M. D., Gordon M. P., Sciaky D., Nester E. W. Octopine and nopaline metabolism in Agrobacterium tumefaciens and crown gall tumor cells: role of plasmid genes. J Bacteriol. 1977 Jan;129(1):101–107. doi: 10.1128/jb.129.1.101-107.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ooms G., Hooykaas P. J., Van Veen R. J., Van Beelen P., Regensburg-Tuïnk T. J., Schilperoort R. A. Octopine Ti-plasmid deletion mutants of agrobacterium tumefaciens with emphasis on the right side of the T-region. Plasmid. 1982 Jan;7(1):15–29. doi: 10.1016/0147-619x(82)90023-3. [DOI] [PubMed] [Google Scholar]
  24. Stachel S. E., An G., Flores C., Nester E. W. A Tn3 lacZ transposon for the random generation of beta-galactosidase gene fusions: application to the analysis of gene expression in Agrobacterium. EMBO J. 1985 Apr;4(4):891–898. doi: 10.1002/j.1460-2075.1985.tb03715.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Stachel S. E., Nester E. W. The genetic and transcriptional organization of the vir region of the A6 Ti plasmid of Agrobacterium tumefaciens. EMBO J. 1986 Jul;5(7):1445–1454. doi: 10.1002/j.1460-2075.1986.tb04381.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Stachel S. E., Nester E. W., Zambryski P. C. A plant cell factor induces Agrobacterium tumefaciens vir gene expression. Proc Natl Acad Sci U S A. 1986 Jan;83(2):379–383. doi: 10.1073/pnas.83.2.379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Stachel S. E., Timmerman B., Zambryski P. 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. 1987 Apr;6(4):857–863. doi: 10.1002/j.1460-2075.1987.tb04831.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Stachel S. E., Zambryski P. C. Agrobacterium tumefaciens and the susceptible plant cell: a novel adaptation of extracellular recognition and DNA conjugation. Cell. 1986 Oct 24;47(2):155–157. doi: 10.1016/0092-8674(86)90437-x. [DOI] [PubMed] [Google Scholar]
  29. Stachel S. E., Zambryski P. C. virA and virG control the plant-induced activation of the T-DNA transfer process of A. tumefaciens. Cell. 1986 Aug 1;46(3):325–333. doi: 10.1016/0092-8674(86)90653-7. [DOI] [PubMed] [Google Scholar]
  30. Tait R. C., Kado C. I. Regulation of the virC and virD promoters of pTiC58 by the ros chromosomal mutation of Agrobacterium tumefaciens. Mol Microbiol. 1988 May;2(3):385–392. doi: 10.1111/j.1365-2958.1988.tb00043.x. [DOI] [PubMed] [Google Scholar]
  31. Veluthambi K., Jayaswal R. K., Gelvin S. B. Virulence genes A, G, and D mediate the double-stranded border cleavage of T-DNA from the Agrobacterium Ti plasmid. Proc Natl Acad Sci U S A. 1987 Apr;84(7):1881–1885. doi: 10.1073/pnas.84.7.1881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Veluthambi K., Ream W., Gelvin S. B. Virulence genes, borders, and overdrive generate single-stranded T-DNA molecules from the A6 Ti plasmid of Agrobacterium tumefaciens. J Bacteriol. 1988 Apr;170(4):1523–1532. doi: 10.1128/jb.170.4.1523-1532.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Wang K., Stachel S. E., Timmerman B., VAN Montagu M., Zambryski P. C. Site-Specific Nick in the T-DNA Border Sequence as a Result of Agrobacterium vir Gene Expression. Science. 1987 Jan 30;235(4788):587–591. doi: 10.1126/science.235.4788.587. [DOI] [PubMed] [Google Scholar]
  34. Winans S. C., Ebert P. R., Stachel S. E., Gordon M. P., Nester E. W. A gene essential for Agrobacterium virulence is homologous to a family of positive regulatory loci. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8278–8282. doi: 10.1073/pnas.83.21.8278. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Yamamoto A., Iwahashi M., Yanofsky M. F., Nester E. W., Takebe I., Machida Y. The promoter proximal region in the virD locus of Agrobacterium tumefaciens is necessary for the plant-inducible circularization of T-DNA. Mol Gen Genet. 1987 Jan;206(1):174–177. doi: 10.1007/BF00326554. [DOI] [PubMed] [Google Scholar]
  36. Yanofsky M. F., Porter S. G., Young C., Albright L. M., Gordon M. P., Nester E. W. The virD operon of Agrobacterium tumefaciens encodes a site-specific endonuclease. Cell. 1986 Nov 7;47(3):471–477. doi: 10.1016/0092-8674(86)90604-5. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES