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Molecular tools for isolate and community studies of Pyrenomycete fungi

Stefan J Green1, Stanley Freeman, Yitzhak Hadar

  • 1Department of Microbiology and Plant Pathology, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, Rehovot, Israel, and Institute of Soil, Water and Environmental Sciences, Agriculture Research Organization, The Volcani Center, P.O. Box 6, Bet-Dagan, 50-250, Israel.

Mycologia
|December 15, 2010
PubMed

Insights

New molecular tools, including specific primers and enhanced ITS-DGGE methods, aid in studying Pyrenomycete fungi. These tools improve the identification and analysis of fungal pathogens in environmental samples.

Area of Science:

  • Mycology
  • Molecular Biology
  • Fungal Genetics

Background:

  • Pyrenomycetes are a diverse monophyletic fungal group, including significant human and plant pathogens.
  • Accurate identification and study of these fungi are crucial for understanding disease dynamics and ecological roles.

Purpose of the Study:

  • To develop novel molecular tools for the specific detection and analysis of Pyrenomycete fungi.
  • To evaluate the sensitivity and applicability of these tools for fungal identification in various sample types.

Main Methods:

  • Sequence analysis of 18S ribosomal DNA (rDNA) to identify conserved and divergent regions.
  • Development and validation of selective PCR primers targeting Pyrenomycetes.
  • Application of PCR and denaturing gradient gel electrophoresis (DGGE) for DNA amplification and analysis.
  • Testing primer specificity using pure cultures and environmental DNA samples.

Main Results:

  • Specific PCR primers were successfully developed for amplifying Pyrenomycete 18S rDNA and internal transcribed spacer (ITS) regions.
  • ITS-DGGE analysis demonstrated higher sensitivity than 18S rDNA DGGE for differentiating closely related fungal isolates.
  • The developed molecular tools were effective in analyzing fungal rDNA from environmental samples.

Conclusions:

  • The study provides valuable molecular tools for Pyrenomycete research, aiding in the study of fungal diversity and pathogenicity.
  • ITS-DGGE offers a sensitive method for high-resolution typing of closely related Pyrenomycete isolates.
  • These advancements facilitate the application of molecular techniques in environmental mycology and pathogen detection.

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