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Interaction of purified DNA with plant protoplasts of different cell cycle stage: The concept of a competent phase for plant cell transformation

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Summary

The polycation mediated attachment of purified tritiated DNA to plant protoplasts has been measured by quantitative microautoradiography. The automated grain counting technique used, also provides information on the cell cycle stage of individual protoplasts, which circumvents the need to synchronize the plant cell population before preparation of protoplasts. With protoplasts from asynchronously dividing suspension cultures of Nicotiana syhestris (NS-1), S-phase protoplasts appear to be inefficient binders of 3H-DNA, as compared with G1 or G2 protoplasts. Protoplasts derived from a tumour line of Crepis capillaris (CAPT) exhibit 3H-DNA binding at all cell cycle phases, but Sphase protoplasts appear to be preferential binders. These differences are discussed with reference to cell cycle kinetics, membrane charge variation and the possibility of increasing the efficiency of genetic transformation of higher plant cells in culture.

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Literature

  • Ashmore, S.E. (1981): Kinetic and cytological analyses of plant tissue cultures. Ph.D. Thesis. Canberra (Australia): Australian National University

    Google Scholar 

  • Ashmore, S.E.; Gould, A.R. (1979): Cell cycle analysis of tumour-derived cultures of Crepis capillaris: a kinetic analogy with proliferation of animal tumour cells. Protoplasma 101, 217–230

    Google Scholar 

  • Ashmore, S.E.; Gould, A.R. (1981): Karyotype evolution in a tumour-derived plant tissue culture analysed by Giemsa C-banding. Protoplasma 106, 297–308

    Google Scholar 

  • Ashmore, S.E; Gould, A.R. (1982): Plant protoplast fusion and the cell cycle. Plant Cell Rep. (In press)

  • Borysenko, J.Z.; Woods, W. (1979): Density distribution and mobility of surface anions on a normal/transformed cell pair. Exp. Cell Res. 118, 215–227

    Google Scholar 

  • Braun, A.C. (1962): Tumour inception and development in the crown gall disease. Ann. Rev. Plant Physiol. 13, 533–558

    Google Scholar 

  • Cress, D.E. (1981): Uptake of plasmid DNA by protoplasts from synchronized soybean cell suspension cultures. Z. Pflanzenphysiol. 105, 467–470

    Google Scholar 

  • Gould, A.R. (1979): Combined microspectrophotometry and automated quantitative autoradiography applied to the analysis of the plant cell cycle. J. Cell Sci. 39, 235–245

    Google Scholar 

  • Gould, A.R.; Ashmore, S.E.; Gibbs, A.J. (1981): Cell cycle related changes in the quantity of TMV virions bound to protoplasts of Nicotiana sylvestris. Protoplasma 108, 211–223

    Google Scholar 

  • Heyn, R.F.; Hermans, A.K.; Schilperoort, A. (1974): Rapid and efficient isolation of highly polymerised plant DNA. Plant Sci. Lett. 2, 73–78

    Google Scholar 

  • Lacks, S.A.(1978): Mechanisms of DNA uptake by cells. In: Genetic interaction and gene transfer. Brookhaven Symp. Biol. 29

  • Noronha-Blob, L.; Pitha, J. (1982): Probe for polyanionic regions on the cell surface. In Vitro 18, 122–128

    Google Scholar 

  • Sompayrac, L.M.; Danna, K.J. (1981): Efficient infection of monkey cells with DNA of simian virus 40. Proc. Natl. Acad. Sci. USA 78, 7575–7578

    Google Scholar 

  • Wigler, M.; Sweet, R.; Sim, G.K.; Wold, B.; Pellicer, A.; Lacy, E.; Maniatis, T.; Silverstein, S.; Axel, R. (1979): Transformation of mammalian cells with genes from procaryotes and eucaryotes. Cell 16, 777–785

    Google Scholar 

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Communicated by P. Maliga

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Gould, A.R., Ashmore, S.E. Interaction of purified DNA with plant protoplasts of different cell cycle stage: The concept of a competent phase for plant cell transformation. Theoret. Appl. Genetics 64, 7–12 (1982). https://doi.org/10.1007/BF00303643

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  • DOI: https://doi.org/10.1007/BF00303643

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