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. 1979 Jul;139(1):280–286. doi: 10.1128/jb.139.1.280-286.1979

R-plasmid-mediated chromosomal gene transfer in Agrobacterium tumefaciens.

S E Hamada, J P Luckey, S K Farrand
PMCID: PMC216856  PMID: 457601

Abstract

Although several techniques are available for transferring the Ti plasmids from one strain of agrobacterium tumefaciens to another, there are no reproducible methods for analysis of chromosomal markers in this phytopathogen. The R plasmid, R68.45, is known to show chromosomal mobilizing ability in several bacterial genera including the closely related Rhizobia. R68.45 was transferred into the prototrophic A. tumefaciens strain 15955. Ten kanamycin-resistant transconjugant clones were tested for chromosomal mobilizing ability by mating with strain SA10, rifampin- and streptomycin-resistant histidine auxotroph of strain 15955. Of the 10 donor clones, 2 showed high chromosomal mobilizing ability. Between 1,000 and 2,000 His+ colony-forming units per ml were obtained, a value 10 to 20 times greater than can be accounted for by spontaneous reversion. Sequential recloning and matings resulted in the isolation of relatively stable donor cultures. Chromosome gene transfer is dependent upon the presence in the donor of R68.45. Donors lacking an R plasmid or harboring the closely related plasmid RP4 failed to yield His+ transconjugants. With strain SA11, a methionine auxotroph of strain SA10, coinheritance of histidine and methionine independence could be demonstrated. Approximately half of the transconjugants also inherited R68.45. These results indicate that A. tumefaciens 15955 is capable of undergoing host chromosomal genetic exchange.

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Selected References

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

  1. Beringer J. E., Hopwood D. A. Chromosomal recombination and mapping in Rhizobium leguminosarum. Nature. 1976 Nov 18;264(5583):291–293. doi: 10.1038/264291a0. [DOI] [PubMed] [Google Scholar]
  2. Chilton M. D., Currier T. C., Farrand S. K., Bendich A. J., Gordon M. P., Nester E. W. Agrobacterium tumefaciens DNA and PS8 bacteriophage DNA not detected in crown gall tumors. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3672–3676. doi: 10.1073/pnas.71.9.3672. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chilton M. D., Drummond M. H., Merio D. J., Sciaky D., Montoya A. L., Gordon M. P., Nester E. W. Stable incorporation of plasmid DNA into higher plant cells: the molecular basis of crown gall tumorigenesis. Cell. 1977 Jun;11(2):263–271. doi: 10.1016/0092-8674(77)90043-5. [DOI] [PubMed] [Google Scholar]
  4. Chilton M. D., Farrand S. K., Levin R., Nester E. W. RP4 promotion of transfer of a large Agrobacterium plasmid which confers virulence. Genetics. 1976 Aug;83(4):609–618. doi: 10.1093/genetics/83.4.609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Genetello C., Van Larebeke N., Holsters M., De Picker A., Van Montagu M., Schell J. Ti plasmids of Agrobacterium as conjugative plasmids. Nature. 1977 Feb 10;265(5594):561–563. doi: 10.1038/265561a0. [DOI] [PubMed] [Google Scholar]
  7. Gibbins A. M., Gregory K. F. Relatedness among Rhizobium and Agrobacterium species determined by three methods of nucleic acid hybridization. J Bacteriol. 1972 Jul;111(1):129–141. doi: 10.1128/jb.111.1.129-141.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Haas D., Holloway B. W. R factor variants with enhanced sex factor activity in Pseudomonas aeruginosa. Mol Gen Genet. 1976 Mar 30;144(3):243–251. doi: 10.1007/BF00341722. [DOI] [PubMed] [Google Scholar]
  9. Holsters M., de Waele D., Depicker A., Messens E., van Montagu M., Schell J. Transfection and transformation of Agrobacterium tumefaciens. Mol Gen Genet. 1978 Jul 11;163(2):181–187. doi: 10.1007/BF00267408. [DOI] [PubMed] [Google Scholar]
  10. Jacob A. E., Cresswell J. M., Hedges R. W., Coetzee J. N., Beringer J. E. Properties of plasmids constructed by the in vitro insertion of DNA from Rhizobium leguminosarum or Proteus mirabilis into RP4. Mol Gen Genet. 1976 Sep 23;147(3):315–323. doi: 10.1007/BF00582883. [DOI] [PubMed] [Google Scholar]
  11. KLEIN D. T., KLEIN R. M. Quantitative aspects of transformation of virulence in Agrobacterium tumefaciens. J Bacteriol. 1956 Sep;72(3):308–313. doi: 10.1128/jb.72.3.308-313.1956. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Levin R. A., Farrand S. K., Gordon M. P., Nester E. W. Conjugation in Agrobacterium tumefaciens in the absence of plant tissue. J Bacteriol. 1976 Sep;127(3):1331–1336. doi: 10.1128/jb.127.3.1331-1336.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Meyers J. A., Sanchez D., Elwell L. P., Falkow S. Simple agarose gel electrophoretic method for the identification and characterization of plasmid deoxyribonucleic acid. J Bacteriol. 1976 Sep;127(3):1529–1537. doi: 10.1128/jb.127.3.1529-1537.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Milani V. J., Heberlein G. T. Transfection in Agrobacterium tumefaciens. J Virol. 1972 Jul;10(1):17–22. doi: 10.1128/jvi.10.1.17-22.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Sciaky D., Montoya A. L., Chilton M. D. Fingerprints of Agrobacterium Ti plasmids. Plasmid. 1978 Feb;1(2):238–253. doi: 10.1016/0147-619x(78)90042-2. [DOI] [PubMed] [Google Scholar]
  16. Sistrom W. R. Transfer of chromosomal genes mediated by plasmid r68.45 in Rhodopseudomonas sphaeroides. J Bacteriol. 1977 Aug;131(2):526–532. doi: 10.1128/jb.131.2.526-532.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Smith E. F., Townsend C. O. A PLANT-TUMOR OF BACTERIAL ORIGIN. Science. 1907 Apr 26;25(643):671–673. doi: 10.1126/science.25.643.671. [DOI] [PubMed] [Google Scholar]
  18. Towner K. J., Vivian A. RP4-mediated conjugation in Acinetobacter calcoaceticus. J Gen Microbiol. 1976 Apr;93(2):355–360. doi: 10.1099/00221287-93-2-355. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Zaenen I., Van Larebeke N., Van Montagu M., Schell J. Supercoiled circular DNA in crown-gall inducing Agrobacterium strains. J Mol Biol. 1974 Jun 15;86(1):109–127. doi: 10.1016/s0022-2836(74)80011-2. [DOI] [PubMed] [Google Scholar]

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