MYCOLab Ai Logo - gold and army green

Optimization of Protoplast Preparation and Establishment of PEG-Mediated Genetic Transformation Method in Cordyceps cicadae

Summary

Scientists successfully developed a method to genetically modify Cordyceps cicadae, a valuable medicinal fungus used in traditional Chinese medicine. By optimizing how to remove the fungus’s protective cell wall and introducing new genes using a technique called PEG-mediated transformation, researchers created a stable system for genetic manipulation. This breakthrough opens the door for improving medicinal compounds in this fungus and advancing its use in treating various health conditions.

Background

Cordyceps cicadae is an important edible medicinal fungus with significant pharmacological properties, but its wild growth and lack of genome annotation have limited the construction of stable genetic transformation systems. The establishment of efficient genetic manipulation methods is essential for industrial exploitation and research of this species.

Objective

To successfully establish an efficient plasmid transformation method in Cordyceps cicadae protoplasts using PEG mediation and optimize protoplast preparation conditions to overcome low transformation efficiency and acquire sufficient protoplasts for transformation.

Results

Optimal protoplast preparation conditions were determined as 2-day-old C. cicadae mycelia with 1.5% lywallzyme hydrolysis at 34°C for 5 hours, yielding 5.1 × 10⁷ CFU/mL protoplasts. Sucrose-containing regeneration medium achieved the highest regeneration rate of 10.43%. PEG-mediated transformation efficiency was 37.3 CFU/µg DNA for pCas9-EGFP and 66.67 CFU/µg DNA for G418 expression cassette.

Conclusion

A robust genetic transformation method for C. cicadae has been successfully established through optimization of protoplast preparation and PEG-mediated transformation. This provides a foundation for further gene editing and metabolic engineering studies in C. cicadae.
  • Published in:Journal of Fungi,
  • Study Type:Experimental Study,
  • Source: PMID: 40137257, DOI: 10.3390/jof11030219
Scroll to Top