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Transformation of Brassica napus L. using Agrobacterium tumefaciens: developmentally regulated expression of a reintroduced napin gene

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Summary

Genetically transformed plants of Brassica napus L. (oilseed rape) were obtained from hypocotyl expiants using Agrobacterium tumefaciens vectors. Hypocotyl explants were inoculated with disarmed or oncogenic A. tumefaciens strains, EHA101 and A281, and then cultured on media containing kanamycin. The A. tumefaciens strains harbored a binary vector, which contained a neomycin phosphotransferase II (NPTII) gene driven by the 35S promoter of cauliflower mosaic virus and an engineered napin (seed storage protein) gene with its own promoter (300 nucleotides 5′ to the start of translation). Transformation of B. napus plants was confirmed by detection of NPT II enzyme activity, Southern blot analysis and inheritance of the kanamycin-resistance trait (NPT II gene) in the progeny. Expression of the engineered napin gene in embryos but not in leaves of transgenic plants was observed by Northern analysis. These data demonstrate that morphologically normal, fertile transgenic B. napus plants can be obtained using Agrobacterium as a gene vector and that developmentally regulated expression of reintroduced genes can be achieved.

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Author notes

  1. M. M. Moloney

    Present address: Department of Biological Sciences, University of Calgary, T2N 1N4, Calgary, Alberta, Canada

Authors and Affiliations

  1. Calgene Inc., 1920 Fifth Street, 95616, Davis, CA, USA

    S. E. Radke, B. M. Andrews, M. M. Moloney, J. C. Kridl & V. C. Knauf

  2. Department of Biology, Indiana University, 47405, Bloomington, IN, USA

    M. L. Crouch

Authors
  1. S. E. Radke
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  2. B. M. Andrews
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  3. M. M. Moloney
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  4. M. L. Crouch
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  5. J. C. Kridl
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  6. V. C. Knauf
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Communicated by I. Potrykus

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Radke, S.E., Andrews, B.M., Moloney, M.M. et al. Transformation of Brassica napus L. using Agrobacterium tumefaciens: developmentally regulated expression of a reintroduced napin gene. Theoret. Appl. Genetics 75, 685–694 (1988). https://doi.org/10.1007/BF00265588

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  • DOI: https://doi.org/10.1007/BF00265588

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