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Isolation of a recombination deficient Agrobacterium tumefaciens mutant

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Summary

The isolation of a recombination deficient (Rec-) strain of Agrobacterium tumefaciens is described. Strain LBA 4011 was mutagenized with nitrosoguanidine and after segregation 18,000 colonies were replica plated and UV irradiated. Twentytwo UV sensitive strains were isolated and tested for methylmethanesulphonate (MMS) sensitivity. Six of these strains were more MMS-sensitive than LBA 4011. A Ti plasmid that was genetically marked with Tn 1 (CbR) was introduced in these strains and the rescue of the CbR marker during superinfection with an incompatible cointegrate plasmid Ti::R 702 was determined. One strain exhibited a large reduction in rescue frequency. It is concluded that the latter strain was recombination deficient. This property did not influence the induction of plant tumours.

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References

  • Bomhoff, G.H., Klapwijk, P.M., Kester, H.C.M., 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. 145, 177–181 (1977)

    Google Scholar 

  • Chilton, M.-D., Drummond, M.H., Merlo, D.J., Sciaky, D., Montoya, A.L., Gordon, M.D., Nester, E.W.: Stable incorporation of plasmid DNA into higher plant cells: The molecular basis of crown gall tumorigenesis. Cell 11, 263–271 (1977)

    Google Scholar 

  • Clark, A.J.: Recombination deficient mutants of E. coli and other bacteria. Annu. Rev. Genet. 7, 67–86 (1973)

    Google Scholar 

  • Datta, N., Hedges, R.W.: Host range of R-factors. J. Gen. Microbiol. 70, 453–460 (1972)

    Google Scholar 

  • Drummond, M.H., Gordon, M.P., Nester, E.W., Chilton, M.-D.: Foreign DNA of bacterial plasmid origin is transcribed in crown gall tumours. Nature 269, 535–536 (1977)

    Google Scholar 

  • Genetello, C., van Larebeke, N., Holsters, M., de Picker, A., van Montagu, M., Schell, J.: Ti plasmids of Agrobacterium as conjugative plasmids. Nautre 265, 561–563 (1977)

    Google Scholar 

  • Hedges, R.W., Jacob, A.E.: Transposition of ampicillin resistance from RP4 to other replicons. Mol. Gen. Genet. 132, 31–40 (1974)

    Google Scholar 

  • Hooykaas, P.J.J., Roobol, C., Schilperoort, R.A.: Transfer of TI plasmids in Agrobacterium tumefaciens. J. Gen. Microbiol 110, 99–109 (1979)

    Google Scholar 

  • Kerr, A., Manigault, P., Tempé, J.: Transfer of virulence in vivo and in vitro in Agrobacterium. Nature 265, 560–561 (1977)

    Google Scholar 

  • Klapwijk, P.M., Hooykaas, P.J.J., Kester, H.C.M., Schilperoort, R.A., Rörsch, A.: Isolation and characterization of Agrobacterium tumefaciens mutants affected in the utilization of octopine, octopinic acid and lysopine. J. Gen. Microbiol. 96, 155–163 (1976)

    Google Scholar 

  • Klapwijk, P.M., de Jonge, A.J.R., Schilperoort, R.A., Rörsch, A.: An enrichment technique for auxotrophs of Agrobacterium tumefaciens using a combination of carbenicillin and lysozyme. J. Gen. Microbiol. 91, 177–182 (1975)

    Google Scholar 

  • Klapwijk, P.M., Scheulderman, T., Schilperoort, R.A.: Co-ordinated regulation of octopine degradation and conjugative transfer of Ti plasmids in Agrobacterium tumefaciens: Evidence for a common regulatory gene and separate operons. J. Bacteriol. 136, 775–785 (1978)

    Google Scholar 

  • Kleckner, N.: Translocatable elements in procaryotes. Cell 11, 11–23 (1977)

    Google Scholar 

  • Larebeke, N. van, Engler, G., Holsters, M., van den Elsacker, S., Zaenen, I., Schilperoort, R.A., Schell, J.: Large plasmid in Agrobacterium tumefaciens essential for crown gall inducing ability. Nature 252, 169–170 (1974)

    Google Scholar 

  • Miller, J.H.: Experiments in microbial genetics. New York: Cold Spring Harbor Laboratory 1972

    Google Scholar 

  • 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. 129, 101–107 (1977)

    Google Scholar 

  • Petit, A., Tempé, J.: Isolation of Agrobacterium Ti-plasmid regulatory mutants. Mol. Gen. Genet. 167, 147–156 (1978)

    Google Scholar 

  • Petit, A., Tempé, J., Kerr, A., Holsters, M., van Montagu, M., Schell, J.: Substrate induction of conjugative activity of Agrobacterium tumefaciens Ti plasmids. Nature 271, 570–571 (1978)

    Google Scholar 

  • Vervliet, G., Holsters, M., Teuchy, H., van Montagu, M., Schell, J.: Characterization of different plaque forming and defective temperate phages in Agrobacterium tumefaciens. J. Gen. Virol. 26, 33–48 (1975)

    Google Scholar 

  • Watson, B., Currier, T.C., Gordon, M.P., Chilton, M.-D., Nester, E.W.: Plasmid required for Virulence of Agrobacterium tumefaciens. J. Bacteriol. 123, 255–264 (1975)

    Google Scholar 

  • Zaenen, I., van, Larebeke, N., Teuchy, H., van Montagu, M., Schell, J.: Supercoiled circular DNA in crown gall inducing Agrobacterium strains. J. Mol. Biol. 86, 109–127 (1974)

    Google Scholar 

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Communicated by J. Schell

This paper forms part of a Ph.D. Thesis submitted at Leiden University by the first author

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Klapwijk, P.M., van Beelen, P. & Schilperoort, R.A. Isolation of a recombination deficient Agrobacterium tumefaciens mutant. Molec. gen. Genet. 173, 171–175 (1979). https://doi.org/10.1007/BF00330307

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  • DOI: https://doi.org/10.1007/BF00330307

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