Plant Systematics and Evolution

, Volume 216, Issue 3–4, pp 243–249

DNA extraction and PCR amplification method suitable for fresh, herbarium-stored, lichenized, and other fungi

  • Oscar F. Cubero
  • Ana Crespo
  • Jamshid Fatehi
  • Paul D. Bridge
Article

Abstract

This paper presents a DNA extraction method suitable for fresh, herbarium-stored, lichenized and other fungal specimens. The method is fast and reliable, comparatively inexpensive and is suitable for obtaining PCR amplification quality DNA from large numbers of samples in a short time. The method has been tested with over 300 samples ofAscochyta, Phyllosticta, Ramalina, Parmelia andPhysconia. Amplifiable fungal DNA was extracted from pure cultures, leaf lesions, whole thalli and dissected “only-fungal” sections of lichenized fungi. In addition, the method has proved suitable for use with herbarium specimens of both lichenized and non-lichenized fungi, stored as dried pure cultures or dried infected plant material.

Key words

Herbarium DNA extraction filamentous fungi lichenized fungi 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Armaleo, D., Clerc, P., 1991: Lichen chimeras; DNA analysis suggests that one fungus forms two morphotypes. — Exp. Mycol.15: 1–10.Google Scholar
  2. —, —, 1995: A rapid and inexpensive method for the purification of DNA from lichens and their symbionts. — Lichenologist27: 207–213.Google Scholar
  3. Bruns, T. D., Fogel, R., Taylor, J. W., 1990: Amplification and sequencing of DNA from fungal herbarium specimens. — Mycologia82: 175–184.Google Scholar
  4. Cooper, A., 1994: DNA from museum specimens. — InHerrman, B., Hummel, S., (Eds): Ancient DNA. — Berlin, Heidelberg, New York: Springer.Google Scholar
  5. Crespo, A., Bridge, P. D., Hawksworth, D. L., 1997: Amplification of fungal rDNA-ITS regions from non-fertile specimens of the lichen-forming genusParmelia. — Lichenologist29: 275–282.Google Scholar
  6. Cubero, O. F., Crespo, A., 1997: Genetic variability in populations ofPhysconia enteroxantha (Nyl.)Poelt. — In: IAL III Abstracts, p. 151. — Salzburg.Google Scholar
  7. Gardes, M., Bruns, T. D., 1993: ITS primers with enhanced specificity for basidiomycetes: application to the identification of mycorrhizae and rust. — Molec. Ecol.2: 113–118.Google Scholar
  8. Grube, M., DePriest, P., Gargas, A., Hafellner, J., 1995: DNA isolation from lichen ascomata. — Mycol. Res.99: 1321–1324.Google Scholar
  9. Höss, M., Pääbo, S., 1993: DNA extraction from Pleistocene bones by a silica-based purification method. — Nucl. Acids Res.21: 3913–3914.Google Scholar
  10. Jones, A. S., 1963: Use of alkyltrimethylammonium bromides for the isolation of ribo- and the deoxyribo-nucleic acids. — Nature199: 280–282.Google Scholar
  11. Lee, S. B., Milgroom, M. G., Taylor, J. W., 1998: A rapid, high yield mini-prep method for isolation of total genomic DNA from fungi. — Fungal Genet. Newslett.35: 23–24.Google Scholar
  12. Pääbo, S., 1990: Amplifying ancient DNA. — InInnis, M. A., Gelfand, D. H., Sninsky, J. J., White, T. J., (Eds): PCR protocols. — New York: Academic Press.Google Scholar
  13. Paterson, R. R. M., Bridge, P. D., 1994: Biochemical techniques for filamentous fungi. — Wallingford: CABI.Google Scholar
  14. Raeder, U., Broda, P., 1985: Rapid preparation of DNA from filamentous fungi. — Lett. Appl. Microbiol.1: 17–20.Google Scholar
  15. Rogers, S. O., Bendich, A. J., 1994: Extraction of total cellular DNA from plants, algae and fungi. — Pl. Molec. Biol. ManualD1: 1–8.Google Scholar
  16. Rehner, S., Bledsoe, C., Mueller, G. J., Ammirati, J. F., 1989: Extraction of DNA from basidiomycetes for ribosomal RNA hybridization. — Canad. J. Bot.67: 1235–1243.Google Scholar
  17. Savolainen, V., Cuénoud, P., Spichiger, R., Martínez, M. D. P., Crèvecoeur, M., Manen, J., 1995: The use of herbarium specimens for DNA phylogenetics; evaluation and improvement. — Pl. Syst. Evol.197: 87–98.Google Scholar
  18. Sambrook, J., Fritsch, E. F., Maniatis, T., 1989: Molecular cloning: a laboratory manual. 2nd edn. — New York: Cold Spring Harbor Laboratory Press.Google Scholar
  19. Taylor, J. W., Swann, E. C., 1994: DNA from herbarium specimens. — InHerrman, B., Hummel, S., (Eds): Ancient DNA. — Berlin, Heidelberg, New York: Springer.Google Scholar
  20. White, T. J., Bruns, T. D., Lee, S., Taylor, J., 1990: Amplification and direct sequencing of fungal ribosomal DNA gens for phylogenetics. — InInnis, M. A., Gelfand, D. H., Sninsky, J. J., White, T. J., (Eds): PCR protocols. — New York: Academic Press.Google Scholar

Copyright information

© Springer-Verlag 1999

Authors and Affiliations

  • Oscar F. Cubero
    • 1
  • Ana Crespo
    • 1
  • Jamshid Fatehi
    • 2
  • Paul D. Bridge
    • 2
  1. 1.Departamento de Biologia Vegetal IIUniversidad ComplutenseMadridSpain
  2. 2.CABI Bioscience UK Centre (Egham)Egham, SurreyUK

Personalised recommendations