Summary
Protoplast-derived colonies of haploid N. plumbaginifolia were selected for by chlorate resistance in media supplemented with casamino acids. Eighty resistant lines were confirmed by a second passage on a higher concentration of chlorate. Frequency of spontaneous mutation ranged from 10−5 to 10−6. Fifty of the resistant lines could be regenerated into plants, and 30 were characterized biochemically. Ninety percent were fully deficient for nitrate reductase activity. The lines were further tested for xanthine dehydrogenase activity and subsequently classified as defective in the apoenzyme (nia type, 26 lines) or the cofactor (cnx type, 4 lines). Two groups had been identified up until now within the cnx type by growth tests on high concentrations of molybdate supplied to the medium. Nitrate reductase deficiency was stably and continously expressed in both variant cell cultures and regenerants. Genetic analysis demonstrated that nitrate reductase deficiency was inherited as a single recessive nuclear gene.
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References
Bourgin JP, Chupeau Y, Missonier C (1979) Plant regeneration from mesophyll protoplasts of several Nicotiana species. Physiol Plant 45:288–292
Bright SWJ, Norbury PB, Wray JL (1982) A conditional-lethal cnx-type nitrate reductase-deficient barley mutant. In: Nitrate assimilation. Int Symp Nitrate Assimilation — molecular and genetic aspects. Abstracts, Gatersleben, GDR, 34 pp
Brock RD, Friedrich EA, Landgridge J (1973) The modification of amino acid composition of higher plants by mutation and selection. In: Nuclear techniques for seed protein improvement. Int Atomic Energy Agency, Vienna, pp 329–338
Durand J (1979) High and reproducible plating efficiencies of protoplasts isolated from in vitro grown haploid N. sylvestris. Z Pflanzenphysiol 93:283–295
Feenstra WJ, Jacobsen E (1980) Isolation of a nitrate reductase deficient mutant of Pisum sativum by means of selection for chlorate resistance. Theor Appl Genet 58: 39–42
Jaworski EG (1971) Nitrate reductase assay in intact plant tissues. Biochem Biophys Res Commun 43:1274–1279
King J, Khanna V (1980) A nitrate reductase less variant isolated from suspension cultures of Datura innoxia. Plant Physiol 66:632–636
Kleinhofs A, Kuo T, Warner RL (1980) Characterization of nitrate reductase-deficient barley mutants. Mol Gen Genet 177:421–425
Marton L, Dury TM, Mendel RR, Maliga P (1982a) Nitrate reductase deficient cell lines from haploid protoplast cultures of Nicotiana plumbaginifolia. Mol Gen Genet 182: 301–304
Marton L, Sidorov V, Biasini G, Maliga P (1982b) Complementation in somatic hybrids indicates four types of nitrate reductase deficient lines in N. plumbaginifolia. Mol Gen Genet 187:1–3
Marzluf GA (1981) Regulation of nitrogen metabolism and gene expression in fungi. Microbiol Rev 45:437–461
Mendel RR, Müller AJ (1976) A common genetic determinant of xanthine dehydrogenase and nitrate reductase in N. tabacum. Biochem Physiol Pflanz 170:538–541
Mendel RR, Alikulov ZA, Lvov NP, Müller AJ (1981) Presence of molybdenum-cofactor in nitrate reductase-deficient mutant cell lines of tobacco. Mol Gen Genet 181:395–399
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio-assay with tobacco tissue culture. Physiol Plant 15:473–479
Murphy TM, Imbrie CW (1981) Induction and characterization of chlorate-resistant strains of Rosa damascena cultured cells. Plant Physiol 67:910–916
Müller AJ, Grafe R (1978) Isolation and characterization of cell lines of N.tabacum lacking nitrate reductase. Mol Gen Genet 161:67–76
Müller AJ (1981) Nitrate reductase mutants and the cell culture approach to higher plant genetics. In: Beliaev DK (ed) Problems in general genetics, vol II, Proc 14th Int Congr Genet, Mir, Moscow, pp 229–237
Müller AJ (1982) Genetic studies on nitrate reduction in Nicotiana and Hordeum vulgare. In: Nitrate assimilation. Int Symp Nitrate Assimilation — molecular and genetic aspects. Abstracts, Gatersleben, GDR, 9 pp
Negrutiu I (1981) Improved conditions for large-scale culture, mutagenesis, and selection of haploid protoplasts of N. plumbaginifolia. Z Pflanzenphysiol 104:431–442
Negrutiu I, Müler JF (1981) Culture conditions of protoplastderived cells of N.sylvestris for mutant selection. Plant Cell Rep 1:14–17
Oostindier-Braaksma FJ, Feenstra WJ (1973) Isolation and characterization of chlorate-resistant mutants of Arabidopsis thaliana. Mutat Res 19:175–185
Strauss A, Bucher F, King PJ (1981) Isolation of biochemical mutants using haploid mesophyll protoplasts of Hyoscyamus muticus. 1. A NO3 − non-utilizing clone. Planta 153: 75–80
Tomsett AB (1982) The genetic of nitrate assimilation in Neurospora crassa. In: Nitrate assimilation. Int Symp Nitrate Assimilation — molecular and genetic aspects. Abstracts, Gatersleben, GDR, 3 pp
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Communicated by G. Melchers
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Negrutiu, I., Dirks, R. & Jacobs, M. Regeneration of fully nitrate reductase — deficient mutants from protoplast culture of Nicotiana plumbaginifolia (Viviani). Theoret. Appl. Genetics 66, 341–347 (1983). https://doi.org/10.1007/BF00251169
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DOI: https://doi.org/10.1007/BF00251169