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High-efficiency transformation system for the biocontrol agents, Trichoderma spp.

A Herrera-Estrella1, G H Goldman, M Van Montagu

  • 1Laboratorium voor Genetica, Rijksuniversiteit Gent, Belgium.

Molecular Microbiology
|May 1, 1990
PubMed
Summary

We developed an efficient transformation system for Trichoderma biocontrol agents using polyethylene glycol and CaCl2. This method yields numerous stable transformants, enhancing genetic manipulation capabilities for these beneficial fungi.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Biotechnology

Background:

  • Trichoderma species are vital biocontrol agents in agriculture.
  • Efficient genetic transformation methods are crucial for improving biocontrol efficacy.
  • Existing transformation protocols for Trichoderma can be inefficient.

Purpose of the Study:

  • To develop an efficient and reliable transformation system for Trichoderma species.
  • To optimize the use of polyethylene glycol (PEG) and calcium chloride (CaCl2) for DNA delivery.
  • To establish stable genetic modifications in Trichoderma for enhanced biocontrol applications.

Main Methods:

  • Utilized polyethylene glycol (PEG) and CaCl2 for protoplast transformation.
  • Employed the plasmid pAN7-1 with a hygromycin-resistance gene as a selectable marker.

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  • Integrated a 2.4kb homologous DNA fragment to ensure transformant stability.

    • Main Results:

    • Achieved high transformation efficiency, yielding 200-800 transformants per microgram of DNA.
    • Observed stable integration of multiple plasmid copies into the Trichoderma genome.
    • Confirmed integration occurred via non-homologous recombination in stable transformants.

    Conclusions:

    • The developed PEG/CaCl2-based system is highly efficient for Trichoderma transformation.
    • This method facilitates the genetic improvement of Trichoderma biocontrol agents.
    • Stable transformants can be generated, paving the way for enhanced agricultural applications.