Summary
Mesophyll protoplasts of the nitrate-reductase (NR)-deficient Nicotiana plumbaginifolia mutant, “Nia26”, were fused with γ-irradiated mesophyll protoplasts of Nicotiana sylvestris, V-42. Hybrid selection was based on complementation of NR deficiency by transfer of the donor NR gene to N. plumbaginifolia. Regenerated hybrids had different numbers of donor chromosomes in a tetraploid background of N. plumbaginifolia. The transfer and expression of different isozymes from the donor were also observed. Six self-fertile regenerants were obtained from 21 independently isolated cell colonies. Progeny analyses revealed: (1) the linkage of NR and shikimate dehydrogenase (ShDh); (2) a stabilization of the transmission rate of NR; and (3) the obtainment of mono- and disomic addition lines in the first and second progeny of the original regenerants. Southern hybridization analyses demonstrated unequivocally the presence of the NR gene from the donor partner in progeny plants.
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Alexander MP (1968) Differential staining of aborted and nonaborted pollen. Stain Technol 44:122–177
Bates GW, Hasenkampf CA, Contolini CL, Piastuch WC (1987) Asymmetric hybridization in Nicotiana by fusion of irradiated protoplasts. Theor Appl Genet 74:718–726
Dellaporta SL, Wood J, Hicks JB (1985) Maize DNA minipreparation. In: Malmberg R, Mesing J, Sussex I (eds) Molecular biology of plants. Cold Spring Harbor Laboratory Press, Cold Spring Harbor/NY, pp 36–37
Dirks R, Negrutiu I, Heinderyckx M, Jacobs M (1986) Genetic analysis of revertants for the nitrate reductase function of Nicotiana plumbaginifolia. Mol Gen Genet 202:309–311
Dudits D, Fejer O, Hadlaczky G, Koncz C, Lazar GB, Horvath G (1980) Intergeneric gene transfer mediated by plant protoplast fusion. Mol Gen Genet 179:283–288
Dudits D, Maroy E, Praznovszky T, Olah Z, Gyorgyey J, Cella R (1987) Transfer of resistance traits from carrot into tobacco by asymmetric somatic hybridization: Regeneration of fertile plants. Proc Natl Acad Sci USA 84:8434–8438
Famelaer I, Cammaerts D, Karp A, Sidorov V, De Brouwer D, Negrutiu I, Jacobs M (1986) Cellular engineering by “gamma-fusion” and “egg transformation”. In: Nuclear techniques and in vitro culture for plant improvement. IAEA, Vienna, pp 453–461
Famelaer I, Gleba YY, Sidorov VA, Kaleda VA, Parakonny AS, Boryshuk NV, Chereb NN, Negrutiu I, Jacobs M (1989) Intrageneric asymmetric hybrids between Nicotiana plumbaginifolia and Nicotiana sylvestris obtained by “gamma-fusion”. Plant Sci 61:105–117
Gleba YY, Hinnisdaels S, Sidorov VA, Kaleda VA, Parakonny AS, Boryshuk NV, Cherep NN, Negrutiu I, Jacobs M (1988) Intergeneric assymmetric hybrids between Nicotiana plumbaginifolia and Atropa belladonna obtained by “gamma-fusion”. Theor Appl Genet 76:760–766
Gupta PP, Schieder O, Gupta M (1984) Intergeneric nuclear gene transfer between somatically and sexually incompatible plants through asymmetric protoplast fusion. Mol Gen Genet 197:30–35
Heijbroek W, Roelands AJ, Jong JH de, Hulst C van, Schoone AHL, Munning RG (1988) Sugar beets homozygous for resistance to beet cyst nematode (Heterodera schachtii Schm), developed from monosomic additions of Beta procumbens to B. vulgaris. Euphytica 38:121–131
Hinnisdaels S, Negrutiu I, Gleba YY, Jacobs M (1989) Plant somatic cell hybridization as a tool for transferring chromosomes between plant species. Meded Fac Landbouwet Rijksuniv Genet 53:1707–1712
Installé P, Negrutiu I, Jacobs M (1985) Protoplast-derived plants in Nicotiana plumbaginifolia Viviani: Improving the regeneration response of wild-type and mutant cultures. J Plant Physiol 119:443–454
Jovin T, Chrambach A, Naughton MA (1964) An apparatus for preparative temperature regulated polyacrylamide gel electrophoresis. Anal Biochem 9:351–369
Kao NN (1982) Plant protoplast fusion and isolation of heterokaryons. In: Wetter LR, Constabel F (eds) Plant tissue culture methods. NRCC 19876, Prairie Reg Laboratories, Saskatoon, pp 49–57
Koornneef M, Den Boer B, Van Loenen-Martinet P, Van Roggen P, Wijbrandi J (1987) Protoplast fusion of kanamycinresistant, cnx-Nicotiana plumbaginifolia with streptomycinresistant N. tabacum (SR1) and the effect of irradiation of the tabacum parent. In: Puite KJ, Dons JJM, Huizing HJ, Kool AJ, Koornneef M, Krens FA (eds) Progress in plant protoplast research. Kluwer, Dordrecht, pp 287–288
Law CN (1983) Chromosome engineering in wheat breeding and its implications for molecular genetic engineering. In: Setlow JW, Hollaender A (eds) Genetic engineering: principles and methods, vol 5. Plenum Press, New York, pp 157–172
Maniatis T, Fritsch EF, Hoffmann NL, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor/NY
Negrutiu I (1989) Improved conditions for large-scale culture, mutagenesis, and selection of haploid protoplasts of Nicotiana plumbaginifolia. Z Pflanzenphysiol 100:373–376
Negrutiu I, Dirks R, Jacobs M (1983) Regeneration of fully nitrate-reductase-deficient mutants from protoplast culture of Nicotiana plumbaginifolia (Viviani). Theor Appl Genet 66:341–347
Pijnacker LP, Ferwarda MA (1984) Giemsa C-banding of potato chromosomes. Can J Genet Cytol 26:415–419
Riley R (1960) The meiotic behavior, fertility, and stability of wheat-rye chromosome addition lines. Heredity 14:89–100
Riley R, Kimber G (1966) The transfer of alien genetic variation to wheat. Rep Plant Breed Inst Cambridge, pp 6–36
Sidorov VA, Zubko MK, Kuchko AA, Komarnitsky IK, Gleba YY (1987) Somatic hybridization in potato: use of γ-irradiated protoplasts of Solanum pinnatisectum in genetic reconstruction. Theor Appl Genet 74:364–368
Vallejos CF (1983) Enzyme activity staining. In: Tanksley SD, Orton TJ (eds) Isozymes in plant genetics and breeding, part B. Elsevier, Amsterdam, p 169
Van Geyt JPCF, Smed E (1984) Polymorphism of some marker enzymes of the sugarbeet (Beta vulgaris L.) investigated by polyacrylamide gel electrophoresis and starch gel electrophoresis. Z Pflanzenzuecht 92:295–308
Vaucheret H, Vincentz M, Kronenberger J, Caboche M, Rouzé P (1989) Molecular cloning and characterization of the two homeologous genes coding for nitrate reductase in tobacco. Mol Gen Genet 216:10–15
Wijbrandi J, Vos JGM, Koornneef M (1987) Transfer of regeneration capacity from Lycopersicum peruvianum to L. esculentum by protoplast fusion. In: Puite KJ, Dons JJM, Huizing HJ, Kool AJ, Koornneef M, Krens FA (eds) Progress in plant protoplast research. Kluwer, Dordrecht, pp 227–230
Zelcer A, Aviv D, Galun E (1978) Interspecific transfer of cytoplasmic male sterility by fusion between protoplasts of normal Nicotiana sylvestris and X-irradiated protoplasts of male sterile N. Tabacum. Z Pflanzenphysiol 90:397–408
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Communicated by Yu. Gleba
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Famelaer, I., Negrutiu, I., Mouras, A. et al. Asymmetric hybridization in Nicotiana by “gamma fusion” and progeny analysis of self-fertile hybrids. Theoret. Appl. Genetics 79, 513–520 (1990). https://doi.org/10.1007/BF00226161
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DOI: https://doi.org/10.1007/BF00226161