Skip to main content
Log in

Efficient induction by amiprophos-methyl and flow-cytometric sorting of micronuclei in Nicotiana plumbaginifolia

  • Published:
Planta Aims and scope Submit manuscript

Abstract

Amiprophos-methyl (APM) is a potential herbicide which acts at the level of microtubules. By exposure of suspension cells of Nicotiana plumbaginifolia to this agent, a high degree of metaphase arrest was observed and single as well as groups of chromosomes were scattered throughout the cell, offering good prospects for application in cytology and chromosome isolation. After prolonged exposure to the drug, the chromosomes decondensed and micronuclei were formed. Based on their DNA content, the micronuclei were sorted by flow cytometry. Prospects for application of isolated micronuclei for partial genome transfer and gene mapping are discussed.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

APM:

amiprophos-methyl

References

  • Barfield, D.G., Robinson, S.J., Shields, R. (1985) Plant regeneration from protoplasts of long term haploid suspension cultures of N. plumbaginifolia. Plant Cell Rep. 4, 104–107

    Google Scholar 

  • De, Laat, A.M.M., Blaas, J. (1984) Flow cytometric characterization and sorting of plant chromosomes. Theor. Appl. Genet. 67, 463–467

    Google Scholar 

  • De Laat, A.M.M., Blaas, J. (1987) An improved method for protoplast microinjection suitable for transfer of entire plant chromosomes. Plant Sci. (in press)

  • Ege, T., Ringertz, N.R. (1974) Preparation of microcells by enucleation of micronucleate cells. Exp. Cell Res. 87, 378–382

    Google Scholar 

  • Ege, T., Ringertz, N.R., Hamberg, H., Sidebottom, E. (1977) Preparation of microcells. Methods Cell Biol. 15, 339–358

    Google Scholar 

  • Eriksson, T. (1966) Partial synchronization of cell division in suspension cultures of Haplopappus gracilis. Physiol. Plant. 19, 900–910

    Google Scholar 

  • Fournier, R.E.K. (1981) A general high effieiency procedure for production of microcell hybrids. Proc. Natl. Acad. Sci. USA 78, 6349–6353

    Google Scholar 

  • Fournier, R.E.K. (1982) Microcell-mediated chromosome transfer. In: Techniques in somatic cell genetics. pp. 309–327, Shay, J.W., ed. Plenum, New York, London

    Google Scholar 

  • Fournier, R.E.K., Juricek, D.K., Ruddle, F.H. (1979) Somatic cell genetic analysis of transgenome integration. Somat. Cell Genet. 5, 1061–1077

    Google Scholar 

  • Fournier, R.E.K., Ruddle, F.H. (1977) Microcell-mediated transfer of murine chromosomes into mouse, Chinese hamster, and human somatic cells. Proc. Natl. Acad. Sci. USA 74, 319–323

    Google Scholar 

  • Gilissen, L.J.W., Hänisch ten Cate, C.H., Keen, B. (1983) A rapid method of determining growth characteristics of plant cell populations in batch suspension culture. Plant Cell Rep. 1, 232–235

    Google Scholar 

  • Kao, K.N., Constabel, F., Michayluk, M.R., Gamborg, O.L. (1974) Plant protoplast fusion and growth of integeneric hybrid cells. Planta 120, 215–227

    Google Scholar 

  • Kiermayer, O., Fedtke, C. (1977) Strong anti-microtubule action of amiprophos-methyl (APM) in Micrasterias. Protoplasma 92, 163–166

    Google Scholar 

  • Koop, H.U., Kiermayer, O. (1980) Protoplasmic streaming in the giant unicellular green alga Acetabularia mediterranea II. Differential sensitivity of movement systems to substances acting on microfilaments and microtubuli. Protoplasma 102, 295–306

    Google Scholar 

  • Malmberg, R.L., Griesbach, R.J. (1980) The isolation of mitotic and meiotic chromosomes from plant protoplasts. Plant Sci. Lett. 17, 141–147

    Google Scholar 

  • McBride, O.W., Peterson, J.L. (1980) Chromosome-mediated gene transfer in mammalian cells. Annu. Rev. Genet. 14, 321–345

    Google Scholar 

  • Morejohn, L.C., Fosket, D.E. (1984) Inhibition of plant microtubule polymerization in vitro by the phosphoric amide herbicide amiprophos-methyl. Science 224, 874–876

    Google Scholar 

  • Phillips, S.G., Phillips, D.M. (1969) Sites of nucleolus production in cultured Chinese hamster cells. J. Cell Biol. 4, 248–268

    Google Scholar 

  • Quader, H., Wagenbreth, I., Robinson, D.G. (1978) Structure, synthesis and orientation of microfibrils. V. On the recovery of Oocystis solitaria from microtubule inhibitor treatment Cytobiologie 18, 39–51

    Google Scholar 

  • Robinson, D.G., Herzog, W. (1977) Structure, synthesis and orientation of microfibrils. III. A survery of the action of microtubule inhibitors on microtubules and microfibril orientation in Oocystis solitaria. Cytobiologie 15, 463–474

    Google Scholar 

  • Sekiguchi, T., Shelton, K., Ringerez, N.R. (1978) DNA content of microcells prepared from rat, kangarroo and mouse cells. Exp. Cell Res. 113, 247–258

    Google Scholar 

  • Sree Ramulu, K., Dijkhuis, P. (1986) Flow cytometric analysis of polysomaty and in vitro genetic instability in potato. Plant Cell Rep. 3, 234–237

    Google Scholar 

  • Sree Ramulu, K., Dijkhuis, P., Hänisch ten Cate, Ch.H., Groot, B. de (1985) Patterns of DNA and chromosome variation during in vitro growth in various genotypes of potato. Plant Sci. 41, 69–78

    Google Scholar 

  • Sumida, S., Ueda, M. (1976) Effect of O-ethyl O-(3-methyl-6-nitrophenyl)N-sec-butyl phosphorothioamidate (S-2846), an experimental herbicide, on mitosis in Allium cepa. Plant Cell Physiol. 17, 1351–1354

    Google Scholar 

  • Szabados, L., Hadalaszky, G., Dudits, D. (1981) Uptake of isolated plant chromosomes by plant protoplasts. Planta 151, 141–145

    Google Scholar 

  • Van der Linden, P.M. (1980) Computer analysis of DNA-histograms, Ph.D. Thesis, State Univ. Utrecht

  • Verhoeven, H.A., de Laat, A.M.M., Blaas, J. (1986) Methodological developments for chromosome-mediated gene transfer in higher plants. In: Biomolecular engineering in the European Community, achievements of the research programme (1982–1986), final rep., pp. 977–981, Magnien, E., ed. Martinus Nijhoff Publishers, Dordrecht

    Google Scholar 

  • Verhoeven, H.A., de Laat, A.M.M., Sree Ramulu, K., Dijkhuis, P., van Vloten-Doting, L. (1987) Induction, isolation and flow cytometric sorting of metaphase chromosomes and micronuclei cronuclei from cell suspension cultures of Nicotiana plumbaginifolia. In: Genetic and cellular engineering of plants and micro-organisms important for agriculture, Commission of the European Communities meeting, Louvain-la-Neuve, March 23–26, pp. 119–120

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

de Laat, A.M.M., Verhoeven, H.A., Sree Ramulu, K. et al. Efficient induction by amiprophos-methyl and flow-cytometric sorting of micronuclei in Nicotiana plumbaginifolia . Planta 172, 473–478 (1987). https://doi.org/10.1007/BF00393862

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00393862

Key words

Navigation