Plant Biol (Stuttg) 2007; 9(4): 541-544
DOI: 10.1055/s-2007-965165
Short Research Paper

Georg Thieme Verlag Stuttgart KG · New York

Genome Size Estimations On Ulmus minor Mill., Ulmus glabra Huds., and Celtis australis L. Using Flow Cytometry

J. Loureiro1 , E. Rodriguez1 , Â. Gomes1 , C. Santos1
  • 1Laboratory of Biotechnology and Cytomics, CESAM, Department of Biology, University of Aveiro, Campus Universitário de Santiago, Aveiro, Portugal
Further Information

Publication History

Received: February 7, 2007

Accepted: March 6, 2007

Publication Date:
20 July 2007 (online)

Abstract

The Ulmaceae family is composed of nearly 2000 species widely distributed in the northern hemisphere. Despite their wide distribution area, there are only four native species in the Iberian Peninsula. In this work the genome size of three of those species (Ulmus minor, U. glabra, and Celtis australis) was estimated using flow cytometry. The nuclear DNA content of C. australis was estimated as 2.46 ± 0.061 pg/2C, of U. minor as 4.25 ± 0.158 pg/2C, and of U. glabra as 4.37 ± 0.103 pg/2C of DNA. No statistically significant differences were detected among individuals of the same species. These species revealed to be problematic for flow cytometric analyses, due to the release of mucilaginous compounds into the nuclear suspension. Despite that, the modified protocol here presented ensured high quality analyses (low coefficient of variation and background debris and nuclear fluorescence stability), opening good perspectives on its application to estimate the genome size of species with similar problems.

References

  • 1 Bennett M. D., Leitch I. J.. Plant DNA C-values database (release 3.0, Dec. 2004). Available at: http://www.rbgkew.org.uk/cval/homepage.html. (2004)
  • 2 Bennett M. D., Leitch I. J.. Genome size evolution in plants. Gregory, T., ed. The Evolution of the Genome. London, UK; Elsevier Academic Press (2005): 89-162
  • 3 Brown S., Bergounioux C., Tallet S., Marie D.. Flow cytometry of nuclei for ploidy and cell cycle analysis. Negrutiu, I. and Gharti-Chherti, G., eds. A Laboratory Guide for Cellular and Molecular Plant Biology. Basel; Bikhauser Verlag (1991): 326-345
  • 4 Doležel J., Bartoš J.. Plant DNA flow cytometry and estimation of nuclear genome size.  Annals of Botany. (2005);  95 99-110
  • 5 Doležel J., Bartoš J., Voglmayr H., Greilhuber J.. Nuclear DNA content and genome size of trout and human.  Cytometry Part A. (2003);  51A 127-128
  • 6 Doležel J., Binarová P., Lucretti S.. Analysis of nuclear DNA content in plant cells by flow cytometry.  Biologia Plantarum. (1989);  31 113-120
  • 7 Doležel J., Göhde W.. Sex determination in dioecious plants Melandrium album and M. rubrum using high resolution flow cytometry.  Cytometry. (1995);  19 103-106
  • 8 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.  Science. (1983);  220 1049-1051
  • 9 Greilhuber J.. Self tanning - a new and important source of stoichiometric error in cytophotometric determination of nuclear DNA content in plants.  Plant Systematics and Evolution. (1988);  158 87-96
  • 10 Loureiro J., Rodriguez E., Doležel J., Santos C.. Comparison of four nuclear isolation buffers for plant DNA flow cytometry.  Annals of Botany. (2006 a);  98 679-689
  • 11 Loureiro J., Rodriguez E., Doležel J., Santos C.. Flow cytometric and microscopic analysis of the effect of tannic acid on plant nuclei and estimation of DNA content.  Annals of Botany. (2006 b);  98 515-527
  • 12 Lysák M., Doležel J.. Estimation of nuclear DNA content in Sesleria (Poaceae).  Caryologia. (1998);  52 123-132
  • 13 Navarro C., Castroviejo S.. Ulmaceae. Castroviejo, S., ed. Flora Ibérica. Madrid; Real Jardín Botánico, CSIC (1995): 244-250
  • 14 Noirot M., Barre P., Duperray C., Louarn J., Hamon S.. Effects of caffeine and chlorogenic acid on propidium iodide accessibility to DNA: consequences on genome size evaluation in coffee tree.  Annals of Botany. (2003);  92 259-264
  • 15 Noirot M., Barre P., Louarn J., Duperray C., Hamon S.. Nucleus-cytosol interactions - a source of stoichiometric error in flow cytometric estimation of nuclear DNA content in plants.  Annals of Botany. (2000);  86 309-316
  • 16 Ohri D., Kumar A.. Nuclear DNA amounts in some tropical hardwoods.  Caryologia. (1986);  39 303-307
  • 17 Otto F.. Preparation and staining of cells for high resolution DNA analysis. Radbruch, A., ed. Flow Cytometry and Cell Sorting. Berlin; Springer-Verlag (1992): 101-104
  • 18 Pfosser M., Amon A., Lelley T., Heberle-Bors E.. Evaluation of sensitivity of flow cytometry in detecting aneuploidy in wheat using disomic and ditelosomic wheat-rye addition lines.  Cytometry. (1995);  21 387-393
  • 19 Richens R.. Elm. Cambridge; Cambridge University Press (1983)
  • 20 Rodriguez E., Gomes A., Loureiro J., Doležel J., Santos C.. Estimation of the genome size of the Iberian Peninsula Ulmaceae. In 9th Iberian Congress of Cytometry, Porto: Book of Abstracts. (2005): pp. 37
  • 21 Stipes R., Campana R.. Compendium of Elm Diseases. St. Paul; American Phytopathological Society (1981): 96pp
  • 22 Suda J.. An employment of flow cytometry into plant biosystematics. PhD Thesis, Charles University in Prague, Prague. Available at: http://www.ibot.cas.cz/fcm/suda/presentation/disertation.pdf. (2004)

J. Loureiro

Department of Biology
University of Aveiro

Campus Universitário de Santiago

3810-193 Aveiro

Portugal

Email: jloureiro@bio.ua.pt

Editor: R. Mendel

    >