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Regeneration of normal and fertile plants that express octopine synthase, from tobacco crown galls after deletion of tumour-controlling functions

Abstract

Crown gall, a neoplasmic transformation of plant cells, is caused by transfer and integration into nuclear plant DNA of T-DNA, a segment of the Ti plasmid of Agrobacterium tumefaciens1–8. In plant cells, this T-DNA is expressed9–12 and determines the morphogenetic growth properties of crown gall cells12–16 and the synthesis of opines17–21. The T-DNA in octopine crown galls is composed of either one or two fragments derived from a continuous Ti sequence. Thus far, all octopine tumour lines contain the TL fragment (14 kilobases (kb)), whereas TR-DNA (6 kb) can be present as an additional sequence22–24. Foreign DNA, inserted in the T region of a Ti plasmid, is co-transferred to the plant genome17,18,21. However, an important requirement for the use of T-DNA as vectors for plant genetic engineering is the regeneration of normal plants which stably maintain, express and sexually transmit the transferred DNA. We report here the isolation, from tumour tissues induced by mutant Ti plasmids, of morphologically normal plants that produce octopine synthase in all tissues. This enzymatic activity is inherited as a dominant mendelian marker25. These regenerated plants retained and expressed only the octopine synthase gene of the T-DNA.

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References

  1. Chilton, M.-D. et al. Cell 11, 263–271 (1977).

    Article  CAS  Google Scholar 

  2. De Beuckeleer, M. et al. in Proc. IVth int. Conf. Plant Pathogenic Bacteria (ed. Station de Phytologie végétale et phytobactériologie) 115–126 (INRA, Angers, 1978).

    Google Scholar 

  3. Schell, J. et al. Proc. R. Soc. B204, 251–266 (1979).

    ADS  CAS  Google Scholar 

  4. Zambryski, P. et al. Science 209, 1385–1391 (1980).

    Article  ADS  CAS  Google Scholar 

  5. Yadav, N. S., Postle, K., Saiki, R. K., Thomashow, M. F. & Chilton, M.-D. Nature 287, 458–461 (1980).

    Article  ADS  CAS  Google Scholar 

  6. Thomashow, M. F. et al. Proc. natn. Acad. Sci. U.S.A. 77, 6448–6452 (1980).

    Article  ADS  CAS  Google Scholar 

  7. Willmitzer, L., De Beuckeleer, M., Lemmers, M., Van Montagu, M. & Schell, J. Nature 287, 359–361 (1980).

    Article  ADS  CAS  Google Scholar 

  8. Chilton, M.-D., Saiki, R. K., Yadav, N., Gordon, M. P. & Quetier, F. Proc. natn. Acad. Sci. U.S.A. 77, 4060–4064 (1980).

    Article  ADS  CAS  Google Scholar 

  9. Drummond, M. H., Gordon, M. P., Nester, E. W. & Chilton, M.-D. Nature 269, 535–536 (1977).

    Article  ADS  CAS  Google Scholar 

  10. Gelvin, S. B., Thomashow, M. F., McPherson, J. C., Gordon, M. P. & Nester, E. W. Proc. natn. Acad. Sci. U.S.A. 79, 76–80 (1982).

    Article  ADS  CAS  Google Scholar 

  11. Willmitzer, L. et al. Molec. gen. Genet. 182, 255–262 (1981).

    Article  CAS  Google Scholar 

  12. Willmitzer, L., Simons, G. & Schell, J. EMBO J. 1, 139–146 (1982).

    Article  CAS  Google Scholar 

  13. Garfinkel, D. J. & Nester, E. W. J. Bact. 144, 732–743 (1980).

    CAS  PubMed  Google Scholar 

  14. Ooms, G., Hooykaas, P. J., Moleman, G. & Schilperoort, R. A. Gene 14, 33–50 (1981).

    Article  CAS  Google Scholar 

  15. De Greve, H., Decraemer, H., Seurinck, J., Van Montagu, M. & Schell, J. Plasmid 6, 235–248 (1981).

    Article  CAS  Google Scholar 

  16. Leemans, J. et al. EMBO J. 1, 147–152 (1982).

    Article  CAS  Google Scholar 

  17. Garfinkel, D. J. et al. Cell 27, 143–153 (1981).

    Article  CAS  Google Scholar 

  18. Hernalsteens, J. P. et al. Nature 287, 654–656 (1980).

    Article  ADS  CAS  Google Scholar 

  19. Holsters, M. et al. Plasmid 3, 212–230 (1980).

    Article  CAS  Google Scholar 

  20. Schröder, J., Schröder, G., Huisman, H., Schilperoort, R. A. & Schell, J. FEBS Lett. 129, 166–168 (1981).

    Article  Google Scholar 

  21. Leemans, J., et al. J. molec. appl. Genet. 1, 149–164 (1981).

    CAS  Google Scholar 

  22. Thomashow, M. F., Nutter, R., Montoya, A. L., Gordon, M. P. & Nester, E. W. Cell 19, 729–739 (1980).

    Article  CAS  Google Scholar 

  23. De Beuckeleer, M., et al. Molec gen. Genet. 183, 283–288 (1981).

    Article  CAS  Google Scholar 

  24. Merlo, D. J. et al. Molec. gen. Genet. 177, 637–643 (1980).

    Article  CAS  Google Scholar 

  25. Otten, L. et al. Molec. gen. Genet. 183, 209–213 (1981).

    Article  CAS  Google Scholar 

  26. Murashige, T. & Skoog, S. Physiologia pl. 15, 473–497 (1962).

    Article  CAS  Google Scholar 

  27. Braun, A. C. & Wood, H. N. Proc. natn. Acad. Sci. U.S.A. 73, 496–500 (1976).

    Article  ADS  CAS  Google Scholar 

  28. Wood, H. N., Binns, A. N. & Braun, A. C. Differentiation 11, 175–180 (1978).

    Article  CAS  Google Scholar 

  29. Binns, A. N., Wood, H. M. & Braun, A. C. Differentiation 19, 97–102 (1981).

    Article  CAS  Google Scholar 

  30. Wullems, G. J., Molendijk, L., Ooms, G. & Schilperoort, R. A. Cell 24, 719–727 (1981).

    Article  CAS  Google Scholar 

  31. Otten, L. A. & Schilperoort, R. A. Biochim. biophys. Acta 527, 497–500 (1978).

    Article  CAS  Google Scholar 

  32. De Greve, H., Dhaese, P., Seurinck, J., Van Montagu, M. & Schell, J. J. molec. appl. Genet. 1, 499–512 (1982).

    CAS  Google Scholar 

  33. Yang, F. & Simpson, R. B. Proc. natn. Acad. Sci. U.S.A. 78, 4151–4155 (1981).

    Article  ADS  CAS  Google Scholar 

  34. De Vos, G., De Beuckeleer, M., Van Montagu, M. & Schell, J. Plasmid 6, 249–253 (1981).

    Article  CAS  Google Scholar 

  35. Wieslander, L. Analyt. Biochem. 98, 305–309 (1979).

    Article  CAS  Google Scholar 

  36. Maniatis, T. et al. Cell 15, 687–701 (1978).

    Article  CAS  Google Scholar 

  37. Holsters, M., Villarroel, R., Van Montagu, M. & Schell, J. Molec. gen. Genet. 185, 283–289 (1982).

    Article  CAS  Google Scholar 

  38. Hohn, B. Meth. Enzym. 68, 299–309 (1979).

    Article  CAS  Google Scholar 

  39. Denhardt, D. T. Biochem. biophys. Res. Commun. 23, 641–646 (1966).

    Article  CAS  Google Scholar 

  40. Southern, E. M. J. molec. Biol. 98, 503–518 (1975).

    Article  CAS  Google Scholar 

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Greve, H., Leemans, J., Hernalsteens, JP. et al. Regeneration of normal and fertile plants that express octopine synthase, from tobacco crown galls after deletion of tumour-controlling functions. Nature 300, 752–755 (1982). https://doi.org/10.1038/300752a0

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