Summary
Nuclei were extracted by crushing formaldehyde-fixed plant tissue, and smeared on slides with chick erythrocytes as internal standards, stained with the DNA-specific fluorochrome DAPI and measured by static cytofluorimetry.
The ratios between the fluorescence of 2 C pea nuclei and of the internal standards corresponded to the ratio of the respective DNA contents reported in the literature; peaks with very good coefficient of variation (5–10%) were obtained and the mean fluorescence of 4 C nuclei was very close to twice that of the 2 C nuclei; percent distribution in G1 or G2 of nuclei from different tissues ofHelianthus andPisum were in full accordance with cytophotometric data in the literature.
These results confirm the reliability of this method of DNA quantification, which is simpler and quicker in comparison with cytophotometric techniques.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bennet MD, Smith JB (1976) Nuclear DNA amounts in Angiosperms. Phil Trans Roy Soc Lond B 274: 227–274
Brunori A, Georgieva J, D'amato F (1970) Further observations on the X-irradiation response of G1 cells in dry seeds. Mutation Res 9: 481–487
Galbraith DW, Hakins KR, Maddox GM, Ayres NM, Sharma DR, Firoozabady E (1983) Rapid flow cytometric analysis of the cell cycle in intact plant tissue. Science 220: 1049–1051
Hamada S, Fujita S (1983) DAPI staining improved for quantitative cytofluorometry. Histochemistry 79: 219–226
Sgorbati S,Levi M,Sparvoli E,Trezzi F,Lucchini G (submitted for publication) Cytometry and flow cytometry of 4′, 6-diamidino-2-phenylindole (DAPI)-stained suspension of nuclei released from fresh and fixed tissues of plants
Slater ML, Sharrow SO, Gart JJ (1977) Cell cycle ofSaccharomyces cerevisiae in populations growing at different rates. Proc Natl Acad Sci USA 74: 3850–3854
Van't Hof J (1974) Control of the cell cycle in higher plants. In:Padilla GM, Camera IL, Zimmerman A Cell cycle controls. Academic Press, New York, pp 77–85
—,Sparrow AH (1963) A relationship between DNA content, nuclear volume and minimum mitotic cycle time. Proc Natl Acad Sci USA 49: 897–902
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Levi, M., Tarquini, F., Sgorbati, S. et al. Determination of DNA content by static cytofluorimetry in nuclei released from fixed plant tissue. Protoplasma 132, 64–68 (1986). https://doi.org/10.1007/BF01275791
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01275791