Providing a toolbox for genomic engineering of Trichoderma aggressivum
- Author: mycolabadmin
- 7/31/2025
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Summary
Scientists have created a set of tools to genetically engineer Trichoderma aggressivum, a fungus that causes green mold disease in cultivated mushrooms but can also be used beneficially. The study provides step-by-step methods for transforming this fungus using both traditional plasmid methods and modern CRISPR gene-editing technology. These tools will help researchers understand how the fungus works and potentially harness its beneficial properties for agriculture.
Background
Trichoderma aggressivum causes green mold disease in commercially cultivated Agaricus bisporus mushrooms, resulting in significant crop losses. However, this fungus also has potential applications as a biocontrol agent and plant growth promoter. Despite its importance, efficient molecular biology tools for T. aggressivum have been lacking.
Objective
To develop and validate transformation protocols and genetic tools for genomic engineering of T. aggressivum. This includes establishing PEG-mediated plasmid transformation and CRISPR-Cas9-based genome editing methods with appropriate selection markers.
Results
Plasmid transformation achieved efficiency of 26.63 cfu/μg DNA using the hph marker. CRISPR-Cas9 successfully disrupted the pyr4 gene, generating uridine auxotrophic strains with four confirmed candidates showing expected phenotypes. The pyr4 gene from T. reesei was successfully established as a functional selection marker in the auxotrophic background.
Conclusion
A comprehensive genetic toolbox for T. aggressivum has been established, including validated transformation protocols and two functional selection markers. These tools enable researchers to investigate secondary metabolite biosynthesis, mycoparasitic mechanisms, and biotechnological applications of T. aggressivum.
- Published in:Microbiology Spectrum,
- Study Type:Methods Development Study,
- Source: PMID: 40744866, DOI: 10.1128/spectrum.00966-25