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Genetic transformation of Medicago truncatula using Agrobacterium with genetically modified Ri and disarmed Ti plasmids

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Summary

Fertile transgenic plants of the annual pasture legume Medicago truncatula were obtained by Agrobacterium-mediated transformation, utilising a disarmed Ti plasmid and a binary vector containing the kanamycin resistance gene under the control of the cauliflower mosaic virus 35S promoter. Factors contributing to the result included an improved plant regeneration protocol and the use of explants from a plant identified as possessing high regeneration capability from tissue culture. Genes present on the T-DNA of the Ri plasmid had a negative effect on somatic embryogenesis. Only tissue inoculated with Agrobacterium strains containing a disarmed Ti plasmid lacking the T-DNA region or a Ri plasmid with an inactivated rol A gene regenerated transgenic plants. Fertile transgenic plants were only obtained with disarmed A. tumefaciens, and the introduced NPT II gene was transmitted to R1 progeny.

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Abbreviations

BAP:

6-benzylaminopurine

NAA:

1-naphthaleneacetic acid

NPT:

neomycin phosphotransferase

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Author notes

  1. M. R. Thomas

    Present address: Division of Horticulture, CSIRO, GPO Box 350, 5001, Adelaide, SA, Australia

Authors and Affiliations

  1. Department of Biological Sciences, The University of Newcastle, 2308, New South Wales, Australia

    M. R. Thomas, R. J. Rose & K. E. Nolan

Authors
  1. M. R. Thomas
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  2. R. J. Rose
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  3. K. E. Nolan
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Additional information

Communicated by G. C. Phillips

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Thomas, M.R., Rose, R.J. & Nolan, K.E. Genetic transformation of Medicago truncatula using Agrobacterium with genetically modified Ri and disarmed Ti plasmids. Plant Cell Reports 11, 113–117 (1992). https://doi.org/10.1007/BF00232161

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  • DOI: https://doi.org/10.1007/BF00232161

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