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Stable and Transient Transformation, and a Promoter Assay in the Selective Lignin-degrading Fungus, Ceriporiopsis subvermispora

Summary

This research developed a new method to modify the genes of an important wood-degrading fungus, Ceriporiopsis subvermispora. This advancement allows scientists to better understand and potentially improve this fungus’s unique abilities. Impact on everyday life: – Could lead to more efficient and environmentally friendly paper production – May improve the conversion of plant waste into biofuels – Could help develop better animal feed from agricultural byproducts – Advances our understanding of how genes work in fungi – May lead to new applications in biotechnology and sustainable manufacturing

Background

Genetic transformation is essential for investigating gene function and molecular breeding in organisms. While transformation systems exist for some basidiomycete fungi, it remains challenging in many species due to issues with marker genes, gene expression signals, DNA integration efficiency, and protoplast regeneration rates. Ceriporiopsis subvermispora is an important selective white rot fungus that degrades lignin without severely degrading cellulose, making it valuable for biopulping and biomass pretreatment applications.

Objective

To develop a genetic transformation system for C. subvermispora and utilize it to analyze promoter sequences, particularly focusing on both stable and transient transformation outcomes.

Results

The study achieved both stable and transient transformation in C. subvermispora. Stable transformants showed random integration of plasmid DNA in the chromosome at different copy numbers. Many transformants showed transient drug resistance that was lost during repeated screening. The minimal promoter region of the gpd gene was determined to be 141 bp, and mutation of the TATA sequence significantly reduced transformation efficiency. The system enabled promoter analysis without complications from chromosomal position effects.

Conclusion

The researchers successfully developed the first transformation system for C. subvermispora, demonstrating both stable chromosomal integration and transient expression. The system proved particularly useful for promoter analysis, allowing quick evaluation of gene expression control sequences without interference from chromosomal position effects. The study identified essential promoter elements and demonstrated the importance of the TATA sequence in gene expression.
  • Published in:AMB Express,
  • Study Type:Laboratory Research,
  • Source: 10.1186/s13568-019-0818-1
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