Abstract
For mass isolation of plant metaphase chromosomes, cultured cells of wheat (Triticum monococcum) and parsley (Petroselinum hortense) were synchronized by hydroxyurea and colchicine treatment. This synchronization procedure resulted in high mitotic synchrony, especially in suspension cultures of parsley in which 80% of the cells were found to be at the metaphase stage. Mitotic protoplasts isolated from these synchronized cell cultures served as a source for isolation of chromosomes. The described isolation and purification method yielded relatively pure chromosome suspension. The uptake of the isolated plant chromosomes into recipient wheat, parsley, and maize protoplasts was induced by polyethylene-glycol treatment. Cytological studies provided evidences for uptake of plant chromosomes into plant protoplasts.
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Abbreviations
- PEG:
-
polyethylene glycol
- HU:
-
hydroxyruea
- C:
-
colchicine
- HUC:
-
hydroxyurea and colchicine
- CIM:
-
chromosome isolation medium
- TCM:
-
Tris chromosome medium
References
Belliard, G., Pelletier, G., Vedel, F., Quatier, F. (1978) Morphological characteristics and chloroplast DNA distribution in different cytoplasmic parasexual hybrids of Nicitiana tabacum. Mol. Gen. Genet. 165, 231–237
Binding, H., Nehls, R. (1978) Somatic cell hybridization of Vicia faba + Petunia hybrida. Mol. Gen. Genet. 164, 137–143
Bonett, H.T., Eriksson, T. (1974) Transfer of algal chloroplasts of higher plants. Planta 120, 71–79
Chen, K., Wildman, S.G., Smith, H.H. (1977) Chloroplast DNA distribution in parasexual hybrids as shown by polypeptide composition of fraction I protein. Proc. Natl. Acad. Sci. USA 74, 5109–5112
Chu, Y., Lark, K.G. (1976) Cell-cycle parameters of soybean (Glycine max L.) cells growing in suspension culture: suitability of the system for genetic studies. Planta 132, 259–268
Cocking, E. (1978) Selection and somatic hybridisation. In: Frontiers of plant tissue culture 1978 pp. 151–158 Thorpe, T.A. ed. University of Calgary
Constabel, F., Kurz, W.G.W., Chatson, K.B., Gamborg, O.L. (1974) Induction of partial synchrony in soybean cell cultures. Exp. Cell Res. 85, 105–110
Davey, M.R., Freason, E.M., Power, J.B. (1976) Polyethylene glycol induced transplantation of chloroplasts into protoplasts: an ultrastructural assessment. Plant Sci. Lett. 7, 7–16
Dudits, D., Kao, K.N., Constabel, F., Gamborg, O.L. (1976) Embryogenesis and formation of tetraploid and hexaploid plants from carrot protoplasts. Can. J. Bot. 54, 1063–1067
Dudits, D., Hadlaczky, Gy., Levi, E., Fejér, O., Haydu, Zs., Lázár, G. (1977) Somatic hybridization of Daucus carota and D. capillifolius by protoplast fusion. Theor. Appl. Genet. 51, 127–132
Dudits, D., Hadlaczky, Gy., Lázár, G., Haydu, Zs. (1980a) Increase in genetic variability through somatic cell hybridization of distantly related plant species. In: Plant cell cultures: Results and perspectives pp. 207–214, Sala, F., Parisi, B., Cella, R., Ciferri, O. eds. Elsevier-North Holland, Amsterdam New York Oxford
Dudits, D., Fejér, O., Hadlaczky, Gy., Koncz, Cs., Lázár, G.B., Horváth, G. (1980b) Intergeneric gene transfer mediated by plant protoplast fusion. Mol. Gen. Genet. 179, 283–288
Eriksson, T. (1966) Partial synchronization of cell division in suspension cultures of Haplopappus gracilis. Physiol. Plant. 19, 900–910
Gamborg, O.L., Eveleigh, D.E. (1968) Culture methods and detection of glycanases in suspension cultures of wheat and barley. Can. J. Biochem. 46, 417–421
Jeppensen, P.G.N., Bankier, A.T., Sanders, L. (1978) Non-histone proteins and the structure of metaphase chromosomes. Exp. Cell Res. 115, 293–302
Kao, K.N. (1977) Chromosomal behaviour in somatic hybrids of soybean and Nicotiana glauca. Mol. Gen. Genet. 150, 225–230
Kao, K.N., Michayluk, M.R. (1974) A method for high-frequency intergeneric fusion of plant protoplasts. Planta 115, 355–367
Lörz, H., Potrykus, I. (1978) Investigation on the transfer of isolated nuclei into plant protoplasts. Theor. Appl. Genet. 53, 251–256
Malmberg, R., Griesbach, R.J. (1980) The isolation of mitotic and meiotic chromosomes from plant protoplasts. Plant Sci. Lett. 17, 141–147
Melchers, G., Sacristan, M.D., Holder, A.A. (1978) Somatic hybrid plants of potato and tomato regenerated from protoplasts. Carlsberg Res. Commun. 43, 203–218
Schieder, O. (1977) Hybridization experiments with protoplasts from chlorophylldeficient mutants of some Solanaceous species. Planta 137, 253–257
Szabados, L., Dudits, D. (1980) Fusion between interphase and mitotic plant protoplasts: induction of premature chromosome condensation. Exp. Cell Res. 127(2), 442–446
Thomas, E., King, P.J., Potrykus, I. (1979) Improvement of crop plants via single cells in vitro an assessment. Z. Pflanzenzücht. 82, 1–30
Wallin, A., Glimelius, K.G., Eriksson, T. (1974) The induction of aggregation and fusion of Daucus carota protoplasts by polyethylene glycol. Z. Pflanzenphysiol. 74, 64–80
Willecke, K. (1978) Results and prospects of chromosomal gene transfer between cultured mammalian cells. Theor. Appl. Genet. 52, 97–104
Wullems, G.J., van der Horst, J., Bootsma, D. (1975) Incorporation of isolated chromosomes and induction of hypoxanthine phosphoribosyltransferase in chinese hamster cells. Somatic Cell Genetics 1, 137–152
Zenkteler, N., Melchers, G. (1978) In vitro hybridization by sexual methods and by fusion of somatic protoplasts. Theor. Appl. Genet. 52, 81–90
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Szabados, L., Hadlaczky, G. & Dudits, D. Uptake of isolated plant chromosomes by plant protoplasts. Planta 151, 141–145 (1981). https://doi.org/10.1007/BF00387815
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DOI: https://doi.org/10.1007/BF00387815