, Volume 165, Issue 1–3, pp 212–215 | Cite as

High-resolution flow cytometry of nuclear DNA in higher plants

  • I. Ulrich
  • W. Ulrich
Rapid Communication


High-resolution flow cytometry of nuclear DNA in higher plants has been performed from chopped plant tissues and plant protoplasts. A preparation and staining procedure with the DNA specific fluorochrome DAPI, successfully employed for precise flow cytometric DNA analysis of animal and human cells has been used in a slightly modified manner for the DNA analysis of plant cell material. High-resolution DNA histograms coefficients of variation about 1–1.5% have been obtained routinely from plant species with different DNA content. Staining of nuclei with DAPI in combination with the protein fluorochrome sulforhodamine 101 allows bi-parametric analysis of nuclear DNA and protein. The described simple and precise method might be very promising for the analysis of DNA in basic and applied cytogenetic investigations of plant cell research.


Higher plants Nuclear DNA Nuclear protein Flow cytometry DAPI SR 101 



coefficient of variation



SR 101

sulforhodamine 101


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Bennett MD, Smith JB (1976) Nuclear DNA amounts in angiosperms. Philos Trans R Soc Lond 274: 227–274Google Scholar
  2. Bergounioux C, Brown SC (1990) Plant cell cycle analysis with isolated nuclei. In: Darzynkiewicz Z, Crissman HA (eds) Methods in cell biology, vol 33. Academic Press, San Diego, pp 563–573Google Scholar
  3. —, Perennes C, Miege C, Gadal P (1983) Rapid flow cytometric analysis of male sterility on protoplast division inPetunia hybrida. Protoplasma 130: 138–144Google Scholar
  4. Dean PN (1987) Data analysis in cell kinetics research. In: Gray JW, Darzynkiewicz Z (eds) Techniques in cell cycle analysis. Humana Press, Clifton, pp 207–253Google Scholar
  5. Galbraith DW (1989) Analysis of higher plants by flow cytometry and cell sorting. Int Rev Cytol 116: 165–228Google Scholar
  6. — (1990) Flow cytometric analysis of plant genoms. In: Darzynkiewicz Z, Crissman HA (eds) Methods in cell biology, vol 33. Academic Press, San Diego, pp 549–562Google Scholar
  7. —, Harkins KR, Maddox JM, Ayres NM, Sharma DP, Firoozabady E (1983) Rapid flow cytometric analysis of the cell cycle in intact plant tissues. Science 220: 1049–1151Google Scholar
  8. Heller FO (1973) DNA-Bestimmungen an Keimwurzelzellen vonVicia faba mit Hilfe der Impulszytophotometrie. Ber Deutsch Bot Ges 86: 437–441Google Scholar
  9. Otto FJ (1990) DAPI staining of fixed cells for high-resolution flow cytometry of nuclear DNA. In: Darzynkiewicz Z, Crissman HA (eds) Method in cell biology, vol 33. Academic Press, San Diego, pp 105–110Google Scholar
  10. —, Oldiges H, Göhde W, Jain VK (1981) Flow cytometric measurement of nuclear DNA variations as a potential in vivo mutagenicity test. Cytometry 2: 189–191Google Scholar
  11. Puite VJ, Ten Broeke WRR (1983) DNA staining of fixed and nonfixed plant protoplasts for flow cytometry with Hoechst 33342. Plant Sci Lett 32: 79–88Google Scholar
  12. Sgorbati S, Lucretti S, Lister A, Moretti F, Brown SC (1988) Biparametric DNA-nuclear protein analysis of quiescent, non-cycling differentiated and cycling cells of plant tissues. Basic Appl Histochem 32: 389Google Scholar
  13. Stöhr M, Vogt-Schaden M, Knobloch M, Vogel R, Futterman G (1978) Evaluation of eight fluorochormes combinations for simultaneous DNA-protein flow analysis. Stain Technol 53: 202–215Google Scholar
  14. Ulrich I, Ulrich W (1988) Flow cytometric DNA-analysis of plant protoplasts stained with DAPI. Z Naturforsch 41 c: 1052–1056Google Scholar
  15. —, Fritz B, Ulrich W (1988) Application of DNA fluorochromes for flow cytometric DNA analysis of plant protoplasts. Plant Sci 55: 151–158Google Scholar
  16. Ulrich W (1990) Aneuploid and polyploid cellular heterogeneity in insect cell material and diptera species analyzed by flow cytometry. Z Naturforsch 45 c: 1027–1030Google Scholar
  17. - (1991) Simultaneous measurement of DAPI-Sulforhodamine 101 stained nuclear DNA and protein in higher plants by flow cytometry. Biotechnic Histochem (in press)Google Scholar

Copyright information

© Springer-Verlag 1991

Authors and Affiliations

  • I. Ulrich
    • 1
  • W. Ulrich
    • 1
  1. 1.Institut für GenetikUniversität HohenheimStuttgart 70Federal Republic of Germany

Personalised recommendations