Improving cellulase production in Trichoderma koningii through RNA interference on ace1 gene expression

Area of Science:

• Molecular Biology
• Mycology
• Biotechnology

Background:

• Filamentous fungi are crucial for industrial enzyme production.
• Gene knockout is challenging in some fungi, limiting strain improvement.
• Ribonucleic acid interference (RNAi) offers sequence-specific gene knockdown potential.

Purpose of the Study:

• To verify the function of the cellulase expression repressor ACE1 in Trichoderma koningii.
• To enhance cellulase and xylanase productivity using RNAi-mediated gene silencing.
• To investigate the regulatory role of ACE1 in gene expression.

Main Methods:

• Construction and transformation of an expression cassette for ACE1-specific double-stranded RNA (dsRNA).
• Selection of transformants with successful ACE1 gene silencing in Trichoderma koningii.
• Analysis of cellulase, xylanase, XYR1, and CRE1 gene expression levels.

Main Results:

• ACE1 gene silencing was successfully achieved in Trichoderma koningii.
• Silencing ACE1 led to elevated expression of cellulase and xylanase genes.
• Downregulation of ACE1 increased XYR1 expression but did not affect CRE1 expression.
• Enhanced production of total proteins, cellulase, and xylanase was observed.

Conclusions:

• ACE1 acts as a repressor of xylanase 1 (XYR1) transcription in Trichoderma koningii.
• ACE1 is not involved in the regulation of CRE1 expression.
• ACE1 is a viable target gene for improving enzyme production in Trichoderma koningii.
• RNAi is an effective tool for enhancing the properties of industrial fungi.