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Stable integration and expression of β-glucuronidase and NPT II genes in mango somatic embryos

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Summary

Somatic proembryos of mango (Mangifera indica L. cv. Hindi) were co-cultivated withAgrobacterium tumefaciens strain A208 harboring pTiT37-Se::pMON 9749 (9749 ASE). Transformed somatic proembryos capable of growing on selection medium containing 200 μg/ml kanamycin produced the characteristic indigo blue precipitate in the presence of 5-bromo-4-chloro-3-glucuronic acid. These proembryos were chimeral consisting of transformed (blue) and nontransformed (yellow/white) cells. A stepwise selection strategy was found necessary to eliminate chimeras. a) Initial screening at 200 μg/ml kanamycin to enable growth of transformed cells, b) further screening at 400 μg/ml kanamycin to reduce chimeras, and c) recovery of pure transformed clones of proembryos in liquid selection medium with 100 μg/ml kanamycin. The integration of the NPT II and GUS genes into mango genome was confirmed by Southern hybridization.

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Author notes

  1. Helena Mathews

    Present address: ILTAB, Division of Plant Biology, The Scripps Research Institute, 10666 North Torrey Pines Road, MRC7, 92037, La Jolla, CA

Authors and Affiliations

  1. Tropical Research and Education Center, University of Florida, 18905 SW 280 Street, 33031, Homestead, Florida

    Helena Mathews & R. E. Litz

  2. Department of Horticulture, University of Georgia, 30602, Athens, Georgia

    Helena Mathews & H. Y. Wetzstein

  3. Department of Forestry, University of Georgia, 30602, Athens, Georgia

    H. D. Wilde & S. A. Merkle

Authors
  1. Helena Mathews
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  2. R. E. Litz
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  3. H. D. Wilde
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  4. S. A. Merkle
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  5. H. Y. Wetzstein
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Mathews, H., Litz, R.E., Wilde, H.D. et al. Stable integration and expression of β-glucuronidase and NPT II genes in mango somatic embryos. In Vitro Cell.Dev.Biol.–Plant 28, 172–178 (1992). https://doi.org/10.1007/BF02823312

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  • DOI: https://doi.org/10.1007/BF02823312

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