Skip to main content
SpringerLink
Log in

Nuclear DNA content of Solanum species grown in vitro, as determined by flow cytometry

  • Published:
Acta Physiologiae Plantarum Aims and scope Submit manuscript

Abstract

The DNA relative content in nuclei from several Solanum species, which were used as partners for somatic hybridization, were determined using a flow cytometry method. The nuclei were isolated mechanically or via protoplasts from leaves of in vitro grown plants. In the case of S. nigrum as well as S. tuberosum cv. Bzura and dihaploid clone H8105, the nuclei were also obtained from suspension cultured cells by lysis of protoplasts. The source and the method of nuclei isolation affected the pattern of nuclear DNA in the genotypes studied. The mesophyll nuclei showed two distinct peaks on the DNA histograms, whereas the DNA peaks produced by cell suspension nuclei were broad and less distinct. The DNA content in the nuclei, calculated from the DNA histograms of the samples and a DNA standard historgam (Trout Red Blood Cells, having DNA content of 5.05 pg per nucleus), were much lower in mesophyll nuclei than in those obtained from the cell suspension for the same genotypes. The results are discussed in respect of the genetic instability of Solanum genotypes. The usefulness of a flow cytometry approach in somatic hybridization research is also discussed.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Avan M.F., Maciejewska U., Kleczkowski K., and Wielgat B. 1997. Differential responses of potato cell suspensions to a culture filtrate of Phytophthora infestans. Z. Naturforsch. 52c: 333–338.

    Google Scholar 

  • Doležel J., Binarova P., and Lucretti S. 1989. Analysis of nuclear DNA content in plant cells by flow cytometry. Biol. Plant. 31: 113–120

    Google Scholar 

  • Doležel J. 1991. Flow cytometry analysis of nuclear DNA content in higher plants. Phytochem. Anal. 2: 143–154

    Article  Google Scholar 

  • Galun E. 1981. Plant protoplasts as physiological tools. Ann. Rev. Plant. Physiol. 32: 237–266

    Article  CAS  Google Scholar 

  • Haberlach G.T., Cohen B.A., Reichert N.A., Baer M.A., Towill L.E. and Helgeson J.P. 1985. Isolation, culture and regeneration of protoplasts from potato and several related Solanum species. Plant Science 39: 67–74.

    Article  Google Scholar 

  • Hanisch ten Cate Ch.H., and Ramulu K.S. 1987. Callus growth, tumor development and polyploidization in the tetraploid potato cultivar Bintje. Plant Sci. 49: 209–216.

    Article  Google Scholar 

  • Jakuczun H., Strzelczyk-Żyta D., Narkiewicz M. 1997. Zastosowanie pośrednich metod oznaczania poziomu ploidalności ziemniaka. Biul. Inst. Ziemniaka 48: 91–98.

    Google Scholar 

  • Menczel L., Nagy F., Kiss Zs.R., and 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.

    Article  CAS  Google Scholar 

  • Menke U., Schilde-Rentschler L., Ruoss B., Zanke C., Hemleben V., Ninnemann H. 1996. Somatic hybrids between the cultivated potato Solanum tuberosum L. and the 1EBN wild species Solanum pinnatisectum Dun.: morphological and molecular characterization. Theor. Appl. Genet., 92: 617–626.

    Article  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Nagy J.I., and Maliga P. 1976. Callus induction and plant regeneration from mesophyll protoplasts of Nicotiana sylvestris. Z. Pflanzenphysiol. 78: 453–455.2

    Google Scholar 

  • Nicoloso F.T., Val J., van der Keur M., van Iren F., and Kijne J.W. 1994. Flow-cytometric cell counting and DNA estimation for the study of plant cell population dynamics. Plant Cell, Tissue and Organ Cult. 39: 251–259.

    Article  CAS  Google Scholar 

  • Ramulu K.S., Dijkhuis P., Hanisch ten Cate Ch.H., and Groot B. De. 1985. Patterns of DNA and chromosome variation during in vitro growth in various genotypes of potato. Plant Sci. 41: 69–78.

    Article  CAS  Google Scholar 

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

    Article  Google Scholar 

  • Ramulu K.S., Dijkhuis P., and Roest S. 1989. Patterns of phenotypic and chromosome variation in plants derived from protoplast cultures of monohaploid, dihaploid and diploid genotypes and in somatic hybrids of potato. Plant Sci. 60: 101–110

    Article  Google Scholar 

  • Sgorbati S., Levi M., Sparvoli E., Trezzi F. and Lucchini G. 1986. Cytometry and flow cytometry of 4′, 6-diamidino-2-phenylindole (DAPI) -stained suspensions of nuclei released from fresh and fixed tissues of plants. Physiol.Plant. 68: 471–476.

    Article  CAS  Google Scholar 

  • Ulrich I. and Ulrich W. 1991. High-resolution flow cytometry of nuclear DNA in higher plants. Protoplasma 165: 212–215.

    Article  Google Scholar 

  • Valkonen J.P.T. 1994. Nuclear DNA content of the Solanum spp. in the series Etuberosa as determined by laser flow cytometry. Ann. Appl. Biol. 125: 589–600.

    Article  Google Scholar 

  • Valkonen J.P.T., Watanabe K.N. and Pehu E. 1994. Analysis of correlation between nuclear diploid Solanum brevidens and (di)haploid S. tuberosum. Jpn J. Genet. 69: 525–536.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5a, Warsaw, Poland

    Urszula Maciejewska & Anna Szczerbakowa

  2. Flow Cytometry Lab., Drug Institute, Chełmska 30/34, Warsaw, Poland

    Janusz S. Skierski

Authors
  1. Urszula Maciejewska
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Janusz S. Skierski
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Anna Szczerbakowa
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Maciejewska, U., Skierski, J.S. & Szczerbakowa, A. Nuclear DNA content of Solanum species grown in vitro, as determined by flow cytometry. Acta Physiol Plant 21, 37–43 (1999). https://doi.org/10.1007/s11738-999-0025-9

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11738-999-0025-9

Key words

Search

Navigation