Abstract
Flow cytometry was used to analyse the DNA content of nuclei isolated from intact plant tissues and from callus and cell suspension cultures invitro. Cell nuclei were isolated either mechanically (chopping, syringing) or by a hypotonic lysis of isolated protoplasts. Although both methods gave similar results, a slight shift to lower ploidy levels was observed after protoplast isolation from intact tissues and calli. No differences were observed if the two methods were compared using cell suspension cultures. The results showed that flow cytometry is a rapid method of nuclear DNA content analysis in intact plant tissues and variousin vitro cultures.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Brown, S., Jullien, M., Coutos-Thevenot, P., Muller, P., Renaudin, J. P.: Present developments of flow cytometry in plant biology. - Biol. Cell58:173–178,1986.
De Laat, A. M. M., Göhde, W., Vogelzang, M. J. D. C.: Determination of ploidy of single plants and plant populations by flow cytometry. - Plant Breed.99 : 303–307,1987.
Doležl, J.: FLOWSTAR: A microcomputer program for flow cytometric data manipulation and analysis. - Acta histochem. (submitted to press), 1988.
Fowke, L. C, Constabel, F.: Plant Protoplasts. — CRC Press, Florida 1985.
Galbraith, D. W.: Flow cytometric analysis of the cell cycle. - In :Vasil, I. K. (ed.) : Cell Culture and Somatic Cell Genetics of Plants, Vol. 1. Pp. 765–777. Academic Press, London-New York 1984.
Galbraith, D. W., Harkins, K. R., Maddox, J. M., Ayres, N. M., Sharma, D. P., Firoozabady, E.: Rapid flow cytometric analysis of the cell cycle in intact plant tissues. - Science220 :1049–1051,1983.
Griesbach, R. J., Malmberg, R. L., Carlson, P. S.:An improved technique for the isolation of higher plant chromosomes. - Plant Sci. Lett.24 : 55–60,1982.
Hänisch Ten Cate, C. H.,Sree Ramulu, K.: Callus growth, tumour development and polyploidization in the tetraploid potato cultivar Bintje. - Plant Sci.49: 209–216,1987.
Hänisch Ten Cate, C. H., Sree Ramulu, K., Dukhuis, P., De Groot, B.: Genetic stability of cultured hairy roots induced byAgrobacterium rhizogenes on tuber discs of potato cv. Bintje. - Plant Sci.49:217–222,1987.
Karlsson, S. B., Vasil, I. K.: Growth, cytology and. flow cytometry of embryogenic cell suspension cultures of Panicum maximumJacq. andPennisetum puipureum Schum. - J. Plant Physiol.123 : 211–227,1986.
Melamed, M. R., Mullaney, P. F., Mendelsohn, M. L.: Flow Cytometry and Sorting. - John Wiley, New York 1979.
Muirhead, K. A., Horan, P. K., Poste, G.: Flow cytometry : Present and future. - Bio/Technology3 :337–356,1985.
Remond, I., Brown, S., Germani, E., Benbadis, M. C.: Nuclear DNA content of protoplasts isolated from asynchronous and aphidicolin synchronized meristematic root tip cells ofVigna radiata. Microfluorometric and flow cytometric analysis using Hoechst 33342 and DAPI. - In : Apport de la Cytométrie en Flux en Biologie Cellulaire et Moléculaire. IIIe Colloque Annuel. Pp. 16a. Gif-sur- Yvette 1986.
Sgorbati, S., Levi, M., Sparvoli, E., Trezzi, F., Lucchini, G.: 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 : 471–476,1986.
Shapiro, H. M.: Practical Flow Cytometry. — Alan R. Liss, New York 1985.
Sree Ramulu, K., Dukhuis, P.: Flow cytometric analysis of polysomaty and in virrogenetic instability in potato. - Plant Cell Rep.3: 234–237,1986.
Thorthwaite, J. T., Thomas, R. A.,Russo, J.,Ownby, H., Malinin, G. I., Hornicek, F., Woolley, T. W., Frederick, J.,Malinin, T. I.,Vazquez, D. A.,Seckinger, D.: A review of DNA flow cytometric preparatory and analytical methods. - In: Russo, J. (ed.) : Immunochemistry in Tumor Diagnosis. Pp. 380–398. Martinus Nijhoff Publishers, Dordrecht-Boston-Lancaster 1985.
Van Prooijen-Knegt, A. C., Redi, C. A., Van Der Ploeg, M.: Quantitative aspects of the cytochemical Feulgen-DNA procedure studied on model systems and cell nuclei. - Histochemistry69: 1–17, 1980.
Uijtewaal, B. A.: Ploidy variability in greenhouse cultured and in vitro propagated potato(Solatium tuberosum) monohaploids (2n = x = 12) as determined by flow cytometry. - Plant Cell Rep.6:252–255,1987.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Dpooležel, J., Binarová, P. & Lcretti, S. Analysis of Nuclear DNA content in plant cells by Flow cytometry. Biol Plant 31, 113–120 (1989). https://doi.org/10.1007/BF02907241
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02907241