Summary
Asymmetric nuclear hybrids have been obtained by fusion of cells from a nitrate-reductase deficient mutant of Nicotiana plumbaginifolia (cnx20) and gamma irradiated protoplasts of Atropa belladonna (irradiation doses tested were 10, 30, 50 and 100 krad). The hybrid formation frequency following selection for genotypic complementation in the NR function was in the range of 0.7%–3.7%. Cytogenetic studies demonstrated that all hybrid plants tested possessed multiple (generally tetra- or hexaploid) sets of N. plumbaginifolia (n = 10) chromosomes along with 6–29 Atropa chromosomes (n = 36), some of which were greatly deleted. Besides the cnxA gene (the selection marker), additional material of the irradiated partner was expressed in some of the lines, as shown by analyses of multiple molecular forms of enzymes. Surprisingly, rDNA genes of both parental species were present and amplified in the majority of the hybrids. Whenever studied, the chloroplast DNA in the hybrids was derived from the Nicotiana parent. Regenerants from some lines flowered and were partially fertile. It is concluded that irradiation of cells of the donor parent before fusion can be used to produce highly asymmetric nuclear hybrid plants, although within the dose range tested, the treatment determined the direction of the elimination but not the degree of elimination of the irradiated genome.
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Abbreviations
- Cp:
-
chloroplast
- CsCl/EtBr:
-
cesiumchloride/ethidiumbromide
- EDTA:
-
ethyldiamine tris acetate
- NR:
-
nitrate reductase
- SDS:
-
sodiumdodecylsulphate
- “Gamma-fusion”:
-
is a conventional abbreviation for fusion experiments in which one fusion partner (the donor) is inactivated by sub- or lethal doses of irradiation before fusion with the other (the recipient) partner.
References
Aviv D, Galun E (1980) Restoration of fertility in cytoplasmic male sterile Nicotiana sylvestris by fusion with X-irradiated N. tabacum protoplasts. Theor Appl Genet 58:121–128
Brewbaker JL, Upadkya MD, Makinen Y, MacDonald T (1968) Isoenzymepolymorphism in flowering plants. 3. Gel electrophoretic methods and applications. Physiol Plant 21:930–940
Chien YC, Kao KN, Wetter LR (1982) Chromosome and isozyme studies of Nicotiana tabacum-Glycine max. hybrid cell lines. Theor Appl Genet 62:301–304
Davies BY (1967) Disc electrophoresis. 2. Method and application to human serum proteins. Ann NY Acad Sci 121:404–427
Dudits D, Fejer O, Hadlaczky GY, Koncz CS, Lazar G, 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 offertile plants. Proc Natl Acad Sci USA 84:8434–8438
Gleba YY, Hoffmann F (1979) Arabidobrassica: plant genome engineering by protoplastfusion. Naturwissenschaften 66:547–554
Gleba YY, Momot VP, Okolot AN, Skarzhynskaya MV, Kotov V (1983) Genetic processes in intergeneric hybrids Atropa + Nicotiana. Theor Appl Genet 65:269–276
Gleba YY, Kanevsky IF, Skarzhynskaya MV, Komarnitsky IK, Cherep NN (1986) Hybrids between tobacco crown gall cells and normal somatic cells of Atropa belladonna. Plant Cell Rep 5:394–397
Glimelius K, Fahlesson J, Sjödin K, Sundberg E, Djupsjölacka M, Fellner-Feldegg H, Bonnet HT (1986) Somatic hybridization and cybridization as potential methods for inducing of the gene pools of crops within Brassicaceae and Solanaceae. In: Horn W, Jensen CI, Odenbach W, Schieder O (eds) Genetic manipulation in plant breeding. Proc Int Symp Eucarpia Berlin. Sept 8–13, 1985. de Gruyter, Berlin New York, pp 663–682
Gupta PP, Gupta M, Schieder O (1982) Correction of nitratereductase defect in auxotrophic plants through protoplast mediated intergeneric gene transfer. Mol Gen Genet 188:378–383
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
Imamura J, Saul MW, Potrykus J (1987) X-ray irradiation promoted asymmetric somatic hybridization and molecular analysis of products. Theor Appl Genet 74:445–450
Installe P, Negrutiu I, Jacobs M (1985) Protoplast derived plants in N. plumbaginifolia: improving the regeneration responses of wild type and mutant cultures. J Plant Physiol 119:443–454
Krumbiegel G, Schieder O (1979) Selection of somatic hybrids after fusion of protoplasts from Datura innoxia Mill and Atropa belladonna L. Planta 145:371–376
Menczel L, Galiba G, Nagy F, Maliga P (1982) Effect of radiation dosage on efficiency of chloroplast transfer by protoplast fusion in Nicotiana. Genetics 100:487–495
Menczel L, Nagy F, Lazar G, Maliga P (1983) Transfer of cytoplasmic male sterility by selection for streptomycin resistance after protoplast fusion in Nicotiana. Mol Gen Genet 189:365–369
Negrutiu I, Dirks R, Jacobs M (1983) Regeneration of fully nitrate reductase deficient mutants from protoplast cultures of haploid N. plumbaginifolia. Theor Appl Genet 66:341–347
Negrutiu I, De Brouwer D, Watts JW, Sidorov VA, Dirks R, Jacobs (1986) Fusion of plant protoplasts: a study using auxotrophic mutants of Nicotiana plumbaginifolia, viviani. Theor Appl Genet 72:279–286
Niizeki M, Tanaka M, Akada S, Hirai A, Saito K (1985) Callus formation of somatic hybrids of rice and soybean and characteristics of the hybrid callus. Jpn J Genet 60:81–92
O'Connell MA, Manson MR (1986) Regeneration of somatic hybrid plants formed between Lycopersicon esculentum and Solanum richii. Theor Appl Genet 72:59–65
Potrykus I, Jia J, Lazar GB, Saul M (1984) Hyoscyamus muticus + Nicotiana tabacum fusion hybrids selected via auxotroph complementation. Plant Cell Rep 3:68–71
Pijnacker LP, Walck K, Ferwerda MA (1986) Behaviour of chromosomes in potato leaf tissue cultured in vitro as studied by Bidc-Giemsa labeling. Theor Appl Genet 72:833–839
Sala C, Biasini MG, Morandi C, Nielsen E, Parisi B, Sala F (1985) Selection and nuclear DNA analysis of cell hybrids between Daucus carota and Oryza sativa. J Plant Physiol 118:409–419
Shure M, Wessler S, Fedoroff N (1983) Molecular identification and isolation of the Waxy locus in maize. Cell 35:225–233
Sidorov VA, Zubko MK, Kuchko AA, Komanitsky IK, Gleba YY (1987) Somatic hybridization in potato: use of gammairradiation protoplasts of Solanum pinnatisectum in genetic reconstruction. Theor Appl Genet 74:364–368
Somers DA, Narayanan KR, Kleinhofs A, Cooper-Blauds S, Cocking EC (1986) Immunological evidence for transfer of the barley nitrate reductase structural gene to N. tobacum by protoplast fusion. Mol Gen Genet 204:296–301
Wetter LR, Kao KN (1980) Chromosome and isoenzyme studies on cells derived from protoplast fusion of Nicotiana glauca with Glycine max-Nicotiana glauca cell hybrids. Theor Appl Genet 57:273–276
Zelcer A, Aviv D, Galun E (1978) Interspecific transfer of cytoplasmic male sterility by fusion between protoplasts of normal Nicotiana sylvestris and x-ray irradiated protoplasts of male sterile N. tabacum. Z Pflanzenphysiol 90:397–408
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Communicated by G. Melchers
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Gleba, Y.Y., Hinnisdaels, S., Sidorov, V.A. et al. Intergeneric asymmetric hybrids between Nicotiana plumbaginifolia and Atropa belladonna obtained by “gamma-fusion”. Theoret. Appl. Genetics 76, 760–766 (1988). https://doi.org/10.1007/BF00303523
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DOI: https://doi.org/10.1007/BF00303523