Skip to main content
Plant Physiology logoLink to Plant Physiology
. 1986 Jun;81(2):708–710. doi: 10.1104/pp.81.2.708

Microspectrofluorometric Measurement of Chloroplast DNA in Dividing and Expanding Leaf Cells of Spinacia oleracea

Margaret E Lawrence 1, John V Possingham 1
PMCID: PMC1075408  PMID: 16664887

Abstract

Absolute DNA amounts of individual chloroplasts from mesophyll and epidermal cells of developing spinach leaves were measured by microspectrofluorometry using the DNA-specific stain, 4,6-diamidino-2-phenyl indole, and the bacterium, Pediococcus damnosus, as an internal standard. Values obtained by this method showed that DNA amounts of individual chloroplasts from mesophyll cells fell within a normal distribution curve, although mean DNA amounts changed during leaf development and also differed from the levels in epidermal chloroplasts. There was no evidence in the data of plastids containing either the high or low levels of DNA which would be indicative of discontinuous polyploidy of plastids, or of division occurring in only a small subpopulation of chloroplasts. By contrast, the distribution of nuclear DNA amounts in the same leaf tissues in which cell division was known to be occurring showed a clear bimodal distribution. We consider that the distribution of chloroplast DNA in the plastid population shows that there is no S-phase of chloroplast DNA synthesis, all chloroplasts in the population in young leaf cells synthesize DNA, and all chloroplasts divide.

Full text

PDF
708

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Boffey S. A., Leech R. M. Chloroplast DNA levels and the control of chloroplast division in light-grown wheat leaves. Plant Physiol. 1982 Jun;69(6):1387–1391. doi: 10.1104/pp.69.6.1387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Heinhorst S., Cannon G., Weissbach A. Chloroplast DNA synthesis during the cell cycle in cultured cells of Nicotiana tabacum: inhibition by nalidixic acid and hydroxyurea. Arch Biochem Biophys. 1985 Jun;239(2):475–479. doi: 10.1016/0003-9861(85)90714-3. [DOI] [PubMed] [Google Scholar]
  3. Kinoshita I., Tsuji H. Benzyladenine-induced increase in DNA content per chloroplast in intact bean leaves. Plant Physiol. 1984 Nov;76(3):575–578. doi: 10.1104/pp.76.3.575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Lamppa G. K., Bendich A. J. Changes in Chloroplast DNA Levels during Development of Pea (Pisum sativum). Plant Physiol. 1979 Jul;64(1):126–130. doi: 10.1104/pp.64.1.126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Rose R. J., Cran D. G., Possingham J. V. Distribution of DNA in dividing spinach chloroplasts. Nature. 1974 Oct 18;251(5476):641–642. doi: 10.1038/251641a0. [DOI] [PubMed] [Google Scholar]
  6. Schmitt J. M., Herrmann R. G. Fractionation of cell organelles in silica sol gradients. Methods Cell Biol. 1977;15:177–200. doi: 10.1016/s0091-679x(08)60216-2. [DOI] [PubMed] [Google Scholar]
  7. WALKER P. M. B., YATES H. B. Nuclear components of dividing cells. Proc R Soc Lond B Biol Sci. 1952 Oct 16;140(899):274–299. doi: 10.1098/rspb.1952.0062. [DOI] [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES