Summary
Nitrate reductase deficient (NR-) cell lines were selected indirectly by their resistance to 40 mM chlorate in protoplast cultures of haploidNicotiana plumbaginifolia. Frequency of the chlorate resistant clones was 5.8×10-5 in non-mutagenized cultures, which could be increased up to 25 times by treatment with N-ethyl-N-nitrosourea (NEU) or gamma irradiation.
Out of 136 chlorate resistant clones 29 were fully deficient in nitrate reductase. The rest of the clones contained decreased or normal levels of NR activity (91 and 16 clones, respectively).
Further characterization was carried out in 9 clones which were fully deficient in NR and in 2 clones containing resisdual (0–5%) NR activity. The clones were tentatively classified as defective in the apoenzyme (7 clones including the 2 with residual NR activity) or the cofactor (4 clones) of NR by the xanthine dehydrogenase activity and in vitro enzyme complementation. The cofactor defectives could be further classified into two groups. In one of these (2 clones) the NR activity could be partially restored by unphysiologically high (0.2–1 mM) molybdate in the culture medium. The other two are new types which have not been described in flowering plants.
Plant regeneration was obtained only in the clones which contained residual NR activity.
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Communicated by G. Melchers
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Márton, L., Dung, T.M., Mendel, R.R. et al. Nitrate reductase deficient cell lines from haploid protoplast cultures ofNicotiana plumbaginifolia . Molec. Gen. Genet. 186, 301–304 (1982). https://doi.org/10.1007/BF00729445
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DOI: https://doi.org/10.1007/BF00729445