ArticleScientific reports2021
Generation of Trichoderma harzianum with pyr4 auxotrophic marker by using the CRISPR/Cas9 system.
Article in Scientific reports, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed, 23 citations in OpenAlex.
- Establishment of the Genetic Transformation System ofJournal of fungi (Basel, Switzerland) · 2026Article
- Biocontrol strategies for fungal diseases ofFrontiers in microbiology · 2026Review
- Simply cut out - Combining CRISPR/Cas9 RNPs and transiently selected telomere vectors for marker free-gene deletion inFrontiers in genome editing · 2025Article
- Genome-wide transcriptome profiling reveals molecular response pathways ofFrontiers in microbiology · 2024Article
- Establishment of a CRISPR/Cas9-Mediated Efficient Knockout System ofJournal of fungi (Basel, Switzerland) · 2023Article
- Strain improvement ofFrontiers in microbiology · 2023Review
- Use of CRISPR-Cas tools to engineer Trichoderma species.Microbial biotechnology · 2022Review
- Review
- Network Analysis Reveals Different Cellulose Degradation Strategies AcrossFrontiers in genetics · 2022Article
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Trichoderma harzianum is a filamentous fungus used as a biological control agent for agricultural pests. Genes of this microorganism have been studied, and their applications are patented for use in biofungicides and plant breeding strategies. Gene editing technologies would be of great importance for genetic characterization of this species, but have not yet been reported. This work describes mutants obtained with an auxotrophic marker in this species using the CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)/ Cas (CRISPR-associated) system. For this, sequences for a guide RNA and Cas9 overexpression were inserted via biolistics, and the sequencing approach confirmed deletions and insertions at the pyr4 gene. Phenotypic characterization demonstrated a reduction in the growth of mutants in the absence of uridine, as well as resistance to 5-fluorotic acid. In addition, the gene disruption did not reduce mycoparasitc activity against phytopathogens. Thus, target disruption of the pyr4 gene in T. harzianum using the CRISPR/Cas9 system was demonstrated, and it was also shown that endogenous expression of the system did not interfere with the biological control activity of pathogens. This work is the first report of CRISPR Cas9-based editing in this biocontrol species, and the mutants expressing Cas9 have potential for the generation of useful technologies in agricultural biotechnology.
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