Skip to main content
SpringerLink
Log in

Estimation of nuclear DNA content of plants by flow cytometry

  • Protocol
  • Published:
Plant Molecular Biology Reporter Aims and scope Submit manuscript

An Erratum to this article was published on 01 November 1991

Abstract

A rapid and simple protocol for estimation of nuclear DNA content of plants is described. Suspensions of intact nuclei are prepared either by chopping plant tissues or lysing protoplasts in a MgSO4 buffer, mixed with DNA standards, and stained with propidium iodide in a solution containing DNAase-free RNAase. Fluorescence intensities of the stained nuclei are measured by a flow cytometer. Values for nuclear DNA content are estimated by comparing fluorescence intensities of the nuclei of the test population with those of appropriate internal DNA standards. The same procedure can also be used for rapid determination of ploidy in plant tissues.

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.

Similar content being viewed by others

Abbreviations

BMS:

Black Mexican sweet corn

C:

DNA content of the complement of unreplicated haploid chromosomes

CV:

coefficient of variation

ddH2O:

double-distilled water

FL:

linear integral fluoresence

FS:

forward-angle light scatter

HLN:

human leucocytes

LFL:

logarithmic integral fluorescence

PI:

propidium iodide

PMT:

photomultiplier tube

TRBC:

trout red blood cells

References

  • Arumuganathan, K., and E. D. Earle. 1991. Nuclear DNA content of some important plant species. Plant Mol. Biol. Reporter. 9:210–220.

    Google Scholar 

  • Arumuganathan, K., J. P. Slattery, S. D. Tanksley and E. D. Earle. 1991. Preparation and flow cytometric analysis of metaphase chromosomes of tomato. Theor. Appl. Genet. (in press).

  • Bennett, M. D. and J. B. Smith. 1976. Nuclear DNA amounts in angiosperms. Phil. Trans. R. Soc. Lond. B 274: 227–274.

    Article  CAS  Google Scholar 

  • Bennett, M. D., J. B. Smith and J. S. Heslop-Harrison. 1982. Nuclear DNA amounts in angiosperms. Proc. R. Soc. Lond. B216: 179–199.

    Article  CAS  Google Scholar 

  • Boyum, A. 1977. Separation of lymphocytes, lymphocyte subgroups and monocytes. Lymphology 10:71–76.

    PubMed  CAS  Google Scholar 

  • Cameron, M. J. 1990. Macintosh graphics for the EPICS flow cytometer user. Cytometry 11:916–918.

    Article  PubMed  CAS  Google Scholar 

  • De Laat, A. M. M., W. Göhde and M. J. D. C. Vogelzang. 1987. Determination of ploidy of single plants and plant populations by flow cytometry. Plant Breeding 99: 303–307.

    Article  Google Scholar 

  • De Rocher, E. J., K. R. Harkins, D. W. Galbraith and H. J. Bohnert. 1990. Developmentally regulated systemic endopolyploidy in succulents with small genomes. Science 250: 99–101

    Article  PubMed  Google Scholar 

  • Fahleson, J., J. Dixelius, E. Sundberg, K. Glimelius. 1988. Correlation between flow cytometric determination of nuclear DNA content and chromosome number in somatic hybrids withinBrassicaceae. Plant Cell Reports 7:74–77.

    Article  CAS  Google Scholar 

  • Galbraith, D. W., K. R. Harkins, J. M. Maddox, N. M. Ayres, D. P. Sharma and E. Firoozabady. 1983. Rapid flow cytometric analysis of the cell cycle in intact plant tissues. Science 220:1049–1051.

    Article  CAS  PubMed  Google Scholar 

  • Gray, J. W. and R. G. Langlois. 1986. Chromosome classification and purification using flow cytometry and sorting. Ann. Rev. Biophys. Biophysic. Chem. 15:195–235.

    Article  CAS  Google Scholar 

  • Leutwiler, L. S., B. R. Hough-Evans and E. M. Meyerowitz. 1984. The DNA ofArabidopsis thaliana. Mol. Gen. Genet. 194:15–23.

    Article  CAS  Google Scholar 

  • Laurie, D. A. and M. D. Bennett. 1985. Nuclear DNA content in the generaZea andSorghum. Intergeneric, interspecific and intraspecific variation. Heredity 55:307–313.

    Google Scholar 

  • Messeguer, R., M. W. Ganal, J. C. Steffens and S. D. Tanksley. 1991. Characterization of the level, target sites, and inheritance of cytosine methylation in tomato nuclear DNA. Plant Mol. Biol. 16:759–770.

    Article  Google Scholar 

  • Rayburn, L. A., J. A. Auger, E. A. Benzinger and A. G. Hepburn. 1989. Detection of intraspecific variation inZea mays L. by flow cytometry. J. Exp. Bot. 40:1179–1183.

    Article  CAS  Google Scholar 

  • Rayburn, A. L. and J. A. Auger. 1990. Genome size variation inZea mays ssp. mays adapted to different altitudes. Theor. Appl. Genet. 79: 470–474.

    Article  Google Scholar 

  • Tiersch, T. R., R. W. Chandler, S. S. Wachtel and S. Elias. 1989. Reference standards for flow cytometry and application in comparative studies of nuclear DNA content. Cytometry 10:706–710.

    Article  PubMed  CAS  Google Scholar 

  • van den Engh G., B. Trask, S. Cram and M. Bartholdi. 1984. Preparation of chromosome suspensions for flow cytometry. Cytometry 5:108–117.

    Article  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Plant Breeding and Biometry, Cornell University, 14853-1902, Ithaca, NY, USA

    K. Arumuganathan & E. D. Earle

Authors
  1. K. Arumuganathan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. E. D. Earle
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

An erratum to this article is available at http://dx.doi.org/10.1007/BF02672017.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Arumuganathan, K., Earle, E.D. Estimation of nuclear DNA content of plants by flow cytometry. Plant Mol Biol Rep 9, 229–241 (1991). https://doi.org/10.1007/BF02672073

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02672073

Key words

Search

Navigation