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Spontaneous extensive chromosome elimination in somatic hybrids between somatically congruent species Nicotiana tabacum L. and Atropa belladonna L.

Summary

Mesophyll protoplasts of the kanamycin-resistant nightshade, Atropa belladonna, were fused with mesophyll protoplasts of the phosphinothricin resistant-tobacco, Nicotiana tabacum. A total of 447 colonies resistant to both inhibitors was selected. Most of them regenerated shoots with morphology similar to one of the earlier obtained and described symmetric somatic hybrids Nicotiana + Atropa. However, three colonies (0.2%) regenerated vigorously growing tobacco-like shoots; they readily rooted, and after transfer to soil, developed into normal, fertile plants. Unlike their tobacco parental line, BarD, the obtained plants are resistant to kanamycin [they root normally in the presence of kanamycin (200 mg/1)] and possess activity of neomycin phosphotransferase (NPT II) with the same electrophoretic mobility as the one of the nightshade line. According to Southern blot hybridization analysis carried out with the use of radioactively labeled cloned fragments of the Citrus lemon ribosomal DNA repeat, as well as with Nicotiana plumbaginifolia genus-specific, interspersed repeat Inp, the kanamycin-resistant plants under investigation have only species-specific hybridizing bands from tobacco. Cytological analysis of the chromosome sets shows that plants of all three lines possess 48 large chromosomes similar to Nicotiana tabacum ones (2n = 48), and one small extra chromosome (chromosome fragment) similar to Atropa belladonna ones (2n = 72). Available data allow the conclusion that highly asymmetric, normal fertile somatic hybrids with a whole diploid Nicotiana tabacum genome and only part (not more than 2.8%) of an Atropa belladonna genome have been obtained without any pretreatment of a donor genome, although both these species are somatically congruent.

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References

  • Agoudgil S, Hinnisdaels S, Mouras A, Negrutiu I, Jacobs M (1990) Metabolic complementation for a single gene function associated with partial and total loss of donor DNA in interspecific somatic hybrids. Theor Appl Genet 80:337–342

    Google Scholar 

  • Bates GW, Hasenkampf CA, Contolini CL, Piastuch WC (1987) Asymmetric hybridization in Nicotiana by fusion of irradiated protoplasts. Theor Appl Genet 74:718–726

    Google Scholar 

  • Boutry M, Chua NH (1985) A nuclear gene encoding the beta subunit of the mitochondrial ATP synthase in Nicotiana plumbaginifolia. EMBO J 4:2159–2165

    Google Scholar 

  • Church GM, Gilbert W (1984) Genomic sequencing. Proc Natl Acad Sci USA 81:1991–1995

    Google Scholar 

  • D'Arcy WG (1979) The classification of the Solanaceae. In: Hawkes JG, Lester RN, Skelding AD (eds) The biology and taxonomy of the Solanaceae. Academic Press, London, pp 3–49

    Google Scholar 

  • Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA minipreparation: version 2. Plant Mol Biol Rep 1:19–21

    Google Scholar 

  • Dudits D, Fejer O, Hadlaczky G, Lazar GB, Horvath G (1980) Intergeneric gene transfer mediated by plant protoplast fusion. Mol Gen Genet 202:309–311

    Google Scholar 

  • Famelaer I, Gleba YY, Sidorov VA, Kaleda VA, Parokonny AS, Borisjuk NV, Cherep NN, Negrutiu I, Jacobs M (1989) Intrageneric asymmetric hybrids between Nicotiana plumbaginifolia and Nicotiana sylvestris obtained by “gamma-fusion”. Plant Sci 61:105–117

    Google Scholar 

  • Gleba Y, Sytnik KM (1984) Protoplast fusion. Springer, Berlin, 203 pp

    Google Scholar 

  • Gleba Y, Momot VP, Cherep NN; Skarzhynskaya MV (1982) Intertribal hybrid cell lines of Atropa belladonna + Nicotiana tabacum obtained by cloning individual protoplast fusion products. Theor Appl Genet 62:75–79

    Google Scholar 

  • Gleba Y, Momot VP, Okolot AN, Skarzhynskaya MV, Kotov V (1983) Genetic processes in intergeneric hybrids Atropa + Nicotiana. Theor Appl Genet 65:269–276

    Google Scholar 

  • Gleba Y, Parokonny AS, Kotov V, Negrutiu I, Momot VP (1987) Spatial separation of parental genomes in hybrids of somatic plant cells. Proc Natl Acad Sci USA 84:3709–3713

    Google Scholar 

  • Gleba Y, Hinnisdaels S, Sidorov VA, Kaleda VA, Parokonny AS, Borisjuk NV, Cherep NN, Negrutiu I, Jacobs M (1988) Intergeneric asymmetric hybrids between Nicotiana plumbaginifolia and Atropa belladonna obtained by “gamma-fusion”. Theor Appl Genet 76:760–766

    Google Scholar 

  • 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

    Google Scholar 

  • Kao KN, Michayluk MR (1975) Nutritional requirements for growth of Vicia hajastana cells and protoplasts at a very low population density in liquid media. Planta 126:105–110

    Google Scholar 

  • Kolosha VO, Fodor II (1986) Cloning of Citrus lemon ribosomal DNA repeat. Mol Biol Moscow 20:656–659

    Google Scholar 

  • Kushnir SG, Shlumukov LR, Pogrebnyak HJ, Berger S, Gleba Y (1987) Functional cybrid plants possessing a Nicotiana genome and an Atropa plastome. Mol Gen Genet 209:159–163

    Google Scholar 

  • Kushnir S, Babiychuk E, Bannikova M, Momot V, Komarnitsky I, Cherep N, Gleba Y (1991a Nucleo-cytoplasmic incompatibility in cybrid plants possessing an Atropa genome and a Nicotiana plastome. Mol Gen Genet 225:225–230

    Google Scholar 

  • Kushnir SG, Babiychuk EL, Gleba Y (1991 b) Fertile true nuclear hybrids between Nicotiana tabacum L. and Atropa belladonna L.: a successful hybridization of species belonging to different taxonomic subfamilies. Plant J (in press)

  • Maliga P, Breznovits AS, Marton L, Joo F (1975) Non-Mendelian streptomycin-resistant tobacco mutant with altered chloroplasts and mitochondria. Nature 255:401–402

    Google Scholar 

  • Maniatis T, Fritsch EE, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor/NY

    Google Scholar 

  • Menczel L, Nagy F, Kiss ZR, Maliga P (1981) Streptomycin-resistant and sensitive somatic hybrids of Nicotiana tabacum + Nicotiana knightiana: correlation of resistance to N. tabacum plastids. Theor Appl Genet 59:191–195

    Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497

    Google Scholar 

  • Pijnacker LP, Ferwerda MA (1984) Giemsa C-banding of potato chromosomes. Can J Genet Cytol 26:415–419

    Google Scholar 

  • Reiss B, Sprengel R, Will M, Schaller M (1984) A new sensitive method for qualitative and quantitative assay of neomycin phosphotransferase in crude cell extracts. Gene 30:211–218

    Google Scholar 

  • Wijbrandi J, Wolters AMA, Koorneef M (1990a) Restriction fragment length polymorphism analysis of somatic hybrids between Lycopersicon esculentum and irradiated L. peruvianum: evidence for limited donor genome elimination and extensive chromosome rearrangements. Mol Gen Genet 222:270–277

    Google Scholar 

  • Wijbrandi J, Posthuma A, Kok JM, Rijken R, Vos JGM, Koorneef M (1990b). Asymmetric somatic hybrids between Lycopersicon esculentum and irradiated Lycopersicon peruvianum. Theor Appl Genet 80:305–312

    Google Scholar 

  • Yamashita Y, Terada R, Nishibayashi S, Shimamoto K (1989) Asymmetric somatic hybrids of Brassica: partial transfer of B. campestris genome into B. oleracea by cell fusion. Theor Appl Genet 77:189–194

    Google Scholar 

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Communicated by G. Wenzel

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Babiychuk, E., Kushnir, S. & Gleba, Y.Y. Spontaneous extensive chromosome elimination in somatic hybrids between somatically congruent species Nicotiana tabacum L. and Atropa belladonna L.. Theoret. Appl. Genetics 84, 87–91 (1992). https://doi.org/10.1007/BF00223985

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  • DOI: https://doi.org/10.1007/BF00223985

Key words

  • Somatic cell fusion
  • Asymmetric hybrids
  • Atropa
  • Nicotiana