Skip to main content
SpringerLink
Log in

DNA, RNA, protein and heterochromatin changes during embryo development and germination of soybean (Glycine max L.)

  • Papers
  • Published:
The Histochemical Journal Aims and scope Submit manuscript

Summary

DNA, RNA, protein and heterochromatin were measured cytophotometrically in developing soybean (Glycine max) seeds. The average 2C DNA content for the soybean genome was 2.64 pg. The amounts of nuclear DNA in embryo axes showed no significant change during embryo development, whereas the DNA content in cotyledon nuclei increased significantly from 3.58 pg to 5.49 pg. The number of endopolyploid nuclei increased from 26% to 48% and the DNA content from 4.45 to 5.49 pg after cessation of cell division. The changes in RNA and protein content during embryo development were in general similar to those in DNA content. This can be interpreted that increased DNA levels in soybean cotyledons generated during embryogeny increase the protein synthesizing capacity. During the first 15 days of germination, the number of endopolyploid nuclei in cotyledons declined from 46% to 4%, and this decline is interpreted as DNA degradation providing a ready source of nucleosides and phosphates during early embryo growth. A later decline, however, between 15 and 20 days after germination, was age related similar to leaf senescence, because the percentage of endopolyploid nuclei remained unchanged while the number of non-viable cells increased. In senescing cotyledons, 73% and 80% of RNA and protein but only 20% of DNA were lost, as compared to dormant cotyledons. The heterochromatin (condensed chromatin) measurements indicated that nuclei of metabolically inactive dormant and senescent cotyledon nuclei contained an average of 33% more heterochromatin than nuclei from the green cotyledons of seedlings.

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.

Institutional subscriptions

Similar content being viewed by others

References

  • Aldana, A. B., Fites, R. C. &Pattee, H. E. (1972) Changes in nucleic acids, protein and ribonuclease activity during maturation of peanut seeds.Pl. Cell Physiol. 13, 515–21.

    Google Scholar 

  • Allison, D. C. &Ridolpho, P. (1980) Use of a trypan blue assay to measure the deoxyribonucleic acid content and radioactive labeling of viable cells.J. Histochem. Cytochem. 28, 700–3.

    Google Scholar 

  • Bachmann, K., Harrington, B. A. &Craig, J. P. (1972) Genome size in birds.Chromosoma 37, 405–16.

    Google Scholar 

  • Beevers, L. &Poulson, R. (1972) Protein synthesis in cotyledons ofPisum sativum L. I. Changes in cell-free amino acid incorporation capacity during seed development and maturation.Pl. Physiol. 49, 476–81.

    Google Scholar 

  • Berlyn, G. P., Dhillon, S. S. &Miksche, J. P. (1979) Feulgen cytophotometry of pine nuclei. II. Effect of pectinase used in cell separation.Stain Technol. 54, 201–4.

    Google Scholar 

  • Berlyn, G. P. &Miksche, J. P. (1976)Botanical Microtechnique and Cytochemistry. Ames: Iowa State University Press.

    Google Scholar 

  • Cecich, R. A. &Horner, H. T. (1977) An ultrastructural and microspectrophotometric study of the shoot apex during the initiation of the first leaf in germinatingPinus banksiana.Am. J. Bot. 64, 207–22.

    Google Scholar 

  • Cherry, J. H. (1963) Nucleic acid changes in the storage tissue of seeds during germination.Biochim. Biophys. Acta 68, 193–8.

    Google Scholar 

  • Davies, D. R. (1976) DNA and RNA contents in relation to cell and seed weight inPisum sativum.Pl. Sci. Lett. 7, 17–25.

    Google Scholar 

  • Dhillon, S. S., Berlyn, G. P. &Miksche, J. P. (1977) Requirement of an internal standard for microspectrophotometric measurements of DNA.Am. J. Bot. 64, 117–21.

    Google Scholar 

  • Dhillon, S. S., Berlyn, G. P. &Miksche, J. P. (1978) Nuclear DNA content in populations ofPinus rigida.Am. J. Bot. 65, 192–6.

    Google Scholar 

  • Dhillon, S. S. &Miksche, J. P. (1980a) Evaluation of age related changes in nuclear size, DNA content and heterochromatin.J. Histochem. Cytochem. 28, 606.

    Google Scholar 

  • Dhillon, S. S. &Miksche, J. P. (1980b) DNA changes in senescing leaves evaluated by cytophotometry and DNA-DNA hybridization.Bot. Soc. Am., Misc. Ser. Publn 158, 30.

    Google Scholar 

  • Dhillon, S. S. &Miksche, J. P. (1981) DNA changes during sequential leaf senescence of tobacco (Nicotiana tabacum L.).Physiol. Pl. 51, 291–8.

    Google Scholar 

  • Dhillon, S. S. &Miksche, J. P. (1982) DNA content and heterochromatin variations in various tissues of peanut (Arachis hypogaea).Am. J. Bot. 69, 219–26.

    Google Scholar 

  • Dhillon, S. S., Rake, A. V. &Miksche, J. P. (1980) Reassociation kinetics and cytophotometric characterization of peanut (Arachis hypogaea L.) DNA.Pl. Physiol. 65, 1121–7.

    Google Scholar 

  • Doerschug, E. B., Miksche, J. P. &Palmer, R. G. (1978) DNA content, ribosomal-RNA gene number, and protein content in soybeans.Can. J. Genet. Cytol. 20, 531–8.

    Google Scholar 

  • Duijndam, W. A. L. &Van Duijn, P. (1975) The influence of chromatin compactness on the stoichiometry of the Feulgen-Schiff procedure studied in model films.J. Histochem. Cytochem. 23, 891–900.

    Google Scholar 

  • Goldberg, R. B. (1978) DNA sequence organization in the soybean plant.Biochem. Genet. 16, 45–68.

    Google Scholar 

  • Goldberg, R. B., Hoschek, G., Tam, S. H., Ditta, G. S. &Breidenbach, R. W. (1981) Abundance, diversity, and regulation of mRNA sequence sets in soybean embryogenesis.Devl Biol. 83, 201–17.

    Google Scholar 

  • Gurley, W. B., Hepburn, A. G. &Key, J. L. (1979) Sequence organization of the soybean genome.Biochim. Biophys. Acta 561, 167–83.

    Google Scholar 

  • Harris, J. B., Schaefer, V. G., Dhillon, S. S. & Miksche, J. P. (1979) Nuclear DNA levels in aging leaf tissues — preliminary studies.Annual Scientific Meeting, Gerontological Society, Washington, DC (Abstract).

  • Krul, W. R. (1974) Nucleic acid and protein metabolism of senescing and regenerating soybean cotyledons.Pl. Physiol. 54, 36–40.

    Google Scholar 

  • Leuchtenberger, C. (1958) Quantitative determination of DNA in cells by Feulgen microspectrophotometry. InGeneral Cytochemical Methods, Vol. I (edited byDanielli, J. E.), pp. 219–278. New York: Academic Press.

    Google Scholar 

  • Liu, K. C., Pappelis, A. J. &Courtis, W. S. (1974) Nuclear dry mass and area in epidermal cells of senescing soybean cotyledons.J. exp. Bot. 25, 208–10.

    Google Scholar 

  • Lucas, A. M. &Jamroz, C. (1961)Atlas of Avian Hematology. Agriculture Monograph 25. Washington, D.C.: United States Department of Agriculture.

    Google Scholar 

  • Mazia, D., Brewer, P. A. &Alfert, M. (1953) The cytochemical staining and measurement of protein with mercuric bromophenol blue.Biol. Bull. 104, 57–67.

    Google Scholar 

  • Miksche, J. P., Dhillon, S. S., Berlyn, G. P. &Landauer, K. J. (1979) Nonspecific light loss and intrinsic DNA variation problems associated with Feulgen DNA cytophotometry.J. Histochem. Cytochem. 27, 1377–9.

    Google Scholar 

  • Millerd, A. &Whitfeld, P. R. (1973) Deoxyribonucleic acid and ribonucleic acid synthesis during the cell expansion phase of cotyledon development inVicia faba L.Pl. Physiol. 51, 1005–10.

    Google Scholar 

  • Millerd, A., Simon, M. &Stern, H. (1971) Legumin synthesis in developing cotyledons ofVicia faba L.Pl. Physiol. 48, 419–25.

    Google Scholar 

  • Mumford, P. M. &Pappelis, A. J. (1979) Nuclear degeneration of epidermal cells of senescing soybean cotyledons.J. exp. Bot. 30, 355–60.

    Google Scholar 

  • Nagl, W. (1979) Differential DNA replication in plants: A critical review.Z. Pflanzenphysiol. 95, 283–314.

    Google Scholar 

  • Öpik, H. (1965) The form of nuclei in the storage cells of the cotyledons of germinating seeds ofPhaseolus vulgaris L.Exp. Cell. Res. 38, 517–22.

    Google Scholar 

  • Ritter, C., Distefano, H. S. &Farah, A. (1961) A method for the cytophotometric estimation or ribonucleic acid.J. Histochem. Cytochem. 9, 97–102.

    Google Scholar 

  • Scharfé, A. &Van Parijs, R. (1973) The formation of polyploid cells in ripening cotyledons ofPisum sativum L. in relation to ribosome and protein synthesis.J. exp. Bot. 24, 216–22.

    Google Scholar 

  • Shapiro, H. S. (1970) Deoxyribonucleic acid content per cell of various organisms. InHandbook of Biochemistry — Selected Data for Molecular Biology (edited byStober, H. A.), 2nd edn, pp. H52–60. Cleveland: Chemical Rubber Co.

    Google Scholar 

  • Sparrow, A. H. &Miksche, J. P. (1961) Correlation of nuclear volume and DNA content with higher plant tolerance to chronic radiation.Science 134, 282–3.

    Google Scholar 

  • Spencer, P. W. &Titus, J. S. (1972) Biochemical and enzymatic changes in apple leaf tissue during autumnal senescence.Pl. Physiol. 49, 746–50.

    Google Scholar 

  • Steel, R. G. D. &Torrie, J. H. (1960)Principles and Procedures of Statistics. New York: McGraw-Hill.

    Google Scholar 

  • Udovenko, G. V. &Gogoleva, L. A. (1974) Dynamics of DNA and RNA content in leaves of wheat during ontogenesis.Fizilogiya Rastenii 21, 1076–8.

    Google Scholar 

  • Walbot, V., Brady, T., Clutter, M. &Sussex, I. (1972) Macromolecular synthesis during plant embryogeny: rates of RNA synthesis inPhaseolus coccineus embryos and suspensors.Devl Biol. 29, 104–11.

    Google Scholar 

  • Walbot, V. &Dure, L. S. (1976) Developmental biochemistry of cotton seed embryogenesis and germination. VII. Characterization of the cotton genome.Molec. Biol. 101, 503–36.

    Google Scholar 

  • Walbot, V. &Goldberg, R. (1979) Plant genome organization and its relationship to classical plant genetics. InNucleic Acids in Plants, (edited byHall, T. C. andDavies, J. W.), pp. 3–40. Boca Raton, Florida: CRC Press Inc.

    Google Scholar 

  • Wang, S. M., Lin, C. Y. &Chen, Y. M. (1979) Developmental changes of DNA, RNA and protein in cotyledons and embryonic axis of germinating soybean seeds.Taiwania 24, 115–25.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Botany, North Carolina State University, 27650, Raleigh, North Carolina, U.S.A.

    Sukhraj S. Dhillon & Jerome P. Miksche

Authors
  1. Sukhraj S. Dhillon
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jerome P. Miksche
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Paper No. 8307 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, North Carolina.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Dhillon, S.S., Miksche, J.P. DNA, RNA, protein and heterochromatin changes during embryo development and germination of soybean (Glycine max L.). Histochem J 15, 21–37 (1983). https://doi.org/10.1007/BF01006069

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01006069

Keywords

Search

Navigation