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Application of Membrane and Cell Wall Selective Fluorescent Dyes for Live-Cell Imaging of Filamentous Fungi
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Detection of hyphal fusion in filamentous fungi using differently fluorescence-labeled histones

Christine Rech1, Ines Engh, Ulrich Kück

  • 1Lehrstuhl für Allgemeine und Molekulare Botanik, Ruhr-Universität Bochum, ND7/131, Universitätsstrasse 150, 44780 Bochum, Germany. christine.rech@ruhr-uni-bochum.de

Current Genetics
|October 12, 2007
PubMed

Insights

We developed a new histone-assisted merged fluorescence (HAMF) assay to detect rare fungal cell fusion events. This efficient method uses fluorescently labeled nuclei to identify heterokaryons in Sordaria macrospora and other fungi.

Area of Science:

  • Mycology
  • Cell Biology
  • Genetics

Background:

  • Cell fusion is a fundamental process in fungal life cycles, crucial for vegetative growth and sexual reproduction.
  • Detecting rare hyphal fusion events in filamentous fungi is challenging, limiting our understanding of this process.

Purpose of the Study:

  • To develop a simple, efficient, and broadly applicable method for detecting hyphal fusion in filamentous fungi.
  • To investigate hyphal fusion rates in Sordaria macrospora using a novel assay.

Main Methods:

  • Developed a histone-assisted merged fluorescence (HAMF) assay using reporter vectors encoding fluorescently tagged histone proteins (HH2B-cyan, HH2A-yellow).
  • Utilized Sordaria macrospora as a model system to visualize and quantify heterokaryon formation resulting from hyphal fusion.
  • Tested the assay's sensitivity by analyzing developmental mutants with potentially reduced fusion rates.

Main Results:

  • The HAMF assay successfully detected rare hyphal fusion events by visualizing fluorescently labeled nuclei in heterokaryons.
  • The assay demonstrated its efficiency and applicability in distinguishing between different fungal strains within a mixture.
  • Confirmed the assay's utility in studying fungal developmental mutants with altered fusion phenotypes.

Conclusions:

  • The HAMF assay provides a robust tool for investigating hyphal fusion in filamentous fungi.
  • This method is applicable to a wide range of fungal species, including those with complex reproductive strategies or lacking asexual spores.
  • The assay facilitates deeper insights into the mechanisms and regulation of fungal cell fusion.

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