Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1985 Jul;163(1):341–348. doi: 10.1128/jb.163.1.341-348.1985

Limited-host-range plasmid of Agrobacterium tumefaciens: molecular and genetic analyses of transferred DNA.

M Yanofsky, A Montoya, V Knauf, B Lowe, M Gordon, E Nester
PMCID: PMC219119  PMID: 4008445

Abstract

A tumor-inducing (Ti) plasmid from a strain of Agrobacterium tumefaciens that induces tumors on only a limited range of plants was characterized and compared with the Ti plasmids from strains that induce tumors on a wide range of plants. Whereas all wide-host-range Ti plasmids characterized to date contain closely linked oncogenic loci within a single transferred DNA (T-DNA) region, homology to these loci is divided into two widely separated T-DNA regions on the limited-host-range plasmid. These two plasmid regions, TA-DNA and TB-DNA, are separated by approximately 25 kilobases of DNA which is not maintained in the tumor. The TA-DNA region resembles a deleted form of the wide-host-range TL-DNA and contains a region homologous to the cytokinin biosynthetic gene. However, a region homologous to the two auxin biosynthetic loci of the wide-host-range plasmid mapped within the TB-DNA region. These latter genes play an important role in tumor formation because mutations in these loci result in a loss of virulence on Nicotiana plants. Furthermore, the TB-DNA region alone conferred tumorigenicity onto strains with an intact set of vir genes. Our results suggest that factors within both the T-DNA and the vir regions contribute to the expression of host range in Agrobacterium species. There was a tremendous variation among plants in susceptibility to tumor formation by various A. tumefaciens strains. This variation occurred not only among different plant species, but also among different varieties of plants within the same genus.

Full text

PDF
341

Images in this article

342Image
on p.342
343Image
on p.343
344Image
on p.344
346Image
on p.346
346Image
on p.346

Selected References

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

  1. Akiyoshi D. E., Morris R. O., Hinz R., Mischke B. S., Kosuge T., Garfinkel D. J., Gordon M. P., Nester E. W. Cytokinin/auxin balance in crown gall tumors is regulated by specific loci in the T-DNA. Proc Natl Acad Sci U S A. 1983 Jan;80(2):407–411. doi: 10.1073/pnas.80.2.407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barry G. F., Rogers S. G., Fraley R. T., Brand L. Identification of a cloned cytokinin biosynthetic gene. Proc Natl Acad Sci U S A. 1984 Aug;81(15):4776–4780. doi: 10.1073/pnas.81.15.4776. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Buchholz W. G., Thomashow M. F. Comparison of T-DNA oncogene complements of Agrobacterium tumefaciens tumor-inducing plasmids with limited and wide host ranges. J Bacteriol. 1984 Oct;160(1):319–326. doi: 10.1128/jb.160.1.319-326.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Buchholz W. G., Thomashow M. F. Host range encoded by the Agrobacterium tumefaciens tumor-inducing plasmid pTiAg63 can be expanded by modification of its T-DNA oncogene complement. J Bacteriol. 1984 Oct;160(1):327–332. doi: 10.1128/jb.160.1.327-332.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Currier T. C., Nester E. W. Isolation of covalently closed circular DNA of high molecular weight from bacteria. Anal Biochem. 1976 Dec;76(2):431–441. doi: 10.1016/0003-2697(76)90338-9. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Garfinkel D. J., Nester E. W. Agrobacterium tumefaciens mutants affected in crown gall tumorigenesis and octopine catabolism. J Bacteriol. 1980 Nov;144(2):732–743. doi: 10.1128/jb.144.2.732-743.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Garfinkel D. J., Simpson R. B., Ream L. W., White F. F., Gordon M. P., Nester E. W. Genetic analysis of crown gall: fine structure map of the T-DNA by site-directed mutagenesis. Cell. 1981 Nov;27(1 Pt 2):143–153. doi: 10.1016/0092-8674(81)90368-8. [DOI] [PubMed] [Google Scholar]
  11. Gelvin S. B., Thomashow M. F., McPherson J. C., Gordon M. P., Nester E. W. Sizes and map positions of several plasmid-DNA-encoded transcripts in octopine-type crown gall tumors. Proc Natl Acad Sci U S A. 1982 Jan;79(1):76–80. doi: 10.1073/pnas.79.1.76. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hoekema A., de Pater B. S., Fellinger A. J., Hooykaas P. J., Schilperoort R. A. The limited host range of an Agrobacterium tumefaciens strain extended by a cytokinin gene from a wide host range T-region. EMBO J. 1984 Dec 20;3(13):3043–3047. doi: 10.1002/j.1460-2075.1984.tb02255.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hooykaas P. J., Hofker M., den Dulk-Ras H., Schilperoort R. A. A comparison of virulence determinants in an octopine Ti plasmid, a nopaline Ti plasmid, and an Ri plasmid by complementation analysis of Agrobacterium tumefaciens mutants. Plasmid. 1984 May;11(3):195–205. doi: 10.1016/0147-619x(84)90026-x. [DOI] [PubMed] [Google Scholar]
  14. Johnson R., Guderian R. H., Eden F., Chilton M. D., Gordon M. P., Nester E. W. Detection and quantitation of octopine in normal plant tissue and in crown gall tumors. Proc Natl Acad Sci U S A. 1974 Feb;71(2):536–539. doi: 10.1073/pnas.71.2.536. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Joos H., Inzé D., Caplan A., Sormann M., Van Montagu M., Schell J. Genetic analysis of T-DNA transcripts in nopaline crown galls. Cell. 1983 Apr;32(4):1057–1067. doi: 10.1016/0092-8674(83)90290-8. [DOI] [PubMed] [Google Scholar]
  16. Kao J. C., Perry K. L., Kado C. I. Indoleacetic acid complementation and its relation to host range specifying genes on the Ti plasmid of Agrobacterium tumefaciens. Mol Gen Genet. 1982;188(3):425–432. doi: 10.1007/BF00330044. [DOI] [PubMed] [Google Scholar]
  17. Klee H., Montoya A., Horodyski F., Lichtenstein C., Garfinkel D., Fuller S., Flores C., Peschon J., Nester E., Gordon M. Nucleotide sequence of the tms genes of the pTiA6NC octopine Ti plasmid: two gene products involved in plant tumorigenesis. Proc Natl Acad Sci U S A. 1984 Mar;81(6):1728–1732. doi: 10.1073/pnas.81.6.1728. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Knauf V. C., Panagopoulos C. G., Nester E. W. Comparison of Ti plasmids from three different biotypes of Agrobacterium tumefaciens isolated from grapevines. J Bacteriol. 1983 Mar;153(3):1535–1542. doi: 10.1128/jb.153.3.1535-1542.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Knauf V., Yanofsky M., Montoya A., Nester E. Physical and functional map of an Agrobacterium tumefaciens tumor-inducing plasmid that confers a narrow host range. J Bacteriol. 1984 Nov;160(2):564–568. doi: 10.1128/jb.160.2.564-568.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Koekman B. P., Ooms G., Klapwijk P. M., Schilperoort R. A. Genetic map of an octopine TI-plasmid. Plasmid. 1979 Jul;2(3):347–357. doi: 10.1016/0147-619x(79)90018-0. [DOI] [PubMed] [Google Scholar]
  21. Leemans J., Deblaere R., Willmitzer L., De Greve H., Hernalsteens J. P., Van Montagu M., Schell J. Genetic Identification of functions of TL-DNA transcripts in octopine crown galls. EMBO J. 1982;1(1):147–152. doi: 10.1002/j.1460-2075.1982.tb01138.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lichtenstein C., Klee H., Montoya A., Garfinkel D., Fuller S., Flores C., Nester E., Gordon M. Nucleotide sequence and transcript mapping of the tmr gene of the pTiA6NC octopine Ti-plasmid: a bacterial gene involved in plant tumorigenesis. J Mol Appl Genet. 1984;2(4):354–362. [PubMed] [Google Scholar]
  23. Loper J. E., Kado C. I. Host range conferred by the virulence-specifying plasmid of Agrobacterium tumefaciens. J Bacteriol. 1979 Aug;139(2):591–596. doi: 10.1128/jb.139.2.591-596.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lundquist R. C., Close T. J., Kado C. I. Genetic complementation of Agrobacterium tumefaciens Ti plasmid mutants in the virulence region. Mol Gen Genet. 1984;193(1):1–7. doi: 10.1007/BF00327406. [DOI] [PubMed] [Google Scholar]
  25. Maniatis T., Jeffrey A., Kleid D. G. Nucleotide sequence of the rightward operator of phage lambda. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1184–1188. doi: 10.1073/pnas.72.3.1184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Ooms G., Hooykaas P. J., Moolenaar G., Schilperoort R. A. Grown gall plant tumors of abnormal morphology, induced by Agrobacterium tumefaciens carrying mutated octopine Ti plasmids; analysis of T-DNA functions. Gene. 1981 Jun-Jul;14(1-2):33–50. doi: 10.1016/0378-1119(81)90146-3. [DOI] [PubMed] [Google Scholar]
  27. Ooms G., Klapwijk P. M., Poulis J. A., Schilperoort R. A. Characterization of Tn904 insertions in octopine Ti plasmid mutants of Agrobacterium tumefaciens. J Bacteriol. 1980 Oct;144(1):82–91. doi: 10.1128/jb.144.1.82-91.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Ream L. W., Gordon M. P., Nester E. W. Multiple mutations in the T region of the Agrobacterium tumefaciens tumor-inducing plasmid. Proc Natl Acad Sci U S A. 1983 Mar;80(6):1660–1664. doi: 10.1073/pnas.80.6.1660. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ruvkun G. B., Ausubel F. M. A general method for site-directed mutagenesis in prokaryotes. Nature. 1981 Jan 1;289(5793):85–88. doi: 10.1038/289085a0. [DOI] [PubMed] [Google Scholar]
  30. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  31. Thomashow M. F., Knauf V. C., Nester E. W. Relationship between the limited and wide host range octopine-type Ti plasmids of Agrobacterium tumefaciens. J Bacteriol. 1981 May;146(2):484–493. doi: 10.1128/jb.146.2.484-493.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Thomashow M. F., Nutter R., Montoya A. L., Gordon M. P., Nester E. W. Integration and organization of Ti plasmid sequences in crown gall tumors. Cell. 1980 Mar;19(3):729–739. doi: 10.1016/s0092-8674(80)80049-3. [DOI] [PubMed] [Google Scholar]
  33. Watson B., Currier T. C., Gordon M. P., Chilton M. D., Nester E. W. Plasmid required for virulence of Agrobacterium tumefaciens. J Bacteriol. 1975 Jul;123(1):255–264. doi: 10.1128/jb.123.1.255-264.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. White F. F., Nester E. W. Hairy root: plasmid encodes virulence traits in Agrobacterium rhizogenes. J Bacteriol. 1980 Mar;141(3):1134–1141. doi: 10.1128/jb.141.3.1134-1141.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Willmitzer L., Otten L., Simons G., Schmalenbach W., Schröder J., Schröder G., Van Montagu M., de Vos G., Schell J. Nuclear and polysomal transcripts of T-DNA in octopine crown gall suspension and callus cultures. Mol Gen Genet. 1981;182(2):255–262. doi: 10.1007/BF00269667. [DOI] [PubMed] [Google Scholar]

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

RESOURCES