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Correction of nitrate reductase defect in auxotrophic plant cells through protoplast-mediated intergeneric gene transfers

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Summary

Nitrate reductase-deficient cells of Nicotiana tabacum cv Gatersleben (coded cnx-68) lacking active molybdenum-cofactor were corrected by introducing the genes from Physalis minima and Datura innoxia into NR- genomes. In these itergeneric reconstruction experiments, X-irradiated inactive mesophyll protoplasts of Physalis and Datura were fused separately with the cultured cell protoplasts of cnx-68 Nicotiana. A total of 45 cell colonies, 37 transformed by Physalis and 8 by Datura, were selected from about 1.7×103 heteroplasmic fusion products. The selection of transformants was made by their ability to grow on a medium containing nitrate as the sole nitrogen source. Some of these transformants were further characterized with respect to nitrate reductase, xanthine dehydrogenase and glutamate dehydrogenase activities, chlorate sensitivity, and chlorophyll synthesis. The restoration of nitrate reductase and xanthine dehydrogenase activities confirm the presence of an active form of the molybdenum-cofactor by the expression of introduced genes of Physalis and Datura into the genome of cnx-68 Nicotiana. Such stable transformations via fusion of normal and highly irradiated protoplasts may have a considerable application in higher plants for introducing desirable characters from diverse genomes.

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Authors and Affiliations

  1. Max-Planck-Institut für Züchtungsforschung (Erwin-Baur-Institut), D-5000, Köln 30, Federal Republic of Germany

    Prem P. Gupta, Manju Gupta & Otto Schieder

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  1. Prem P. Gupta
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  2. Manju Gupta
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  3. Otto Schieder
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Communicated by G. Melchers

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Gupta, P.P., Gupta, M. & Schieder, O. Correction of nitrate reductase defect in auxotrophic plant cells through protoplast-mediated intergeneric gene transfers. Mol Gen Genet 188, 378–383 (1982). https://doi.org/10.1007/BF00330037

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  • DOI: https://doi.org/10.1007/BF00330037

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