Related Experiment Video
Updated: Dec 15, 2025
CRISPR-Cas9-Mediated Genome Editing in the Filamentous Ascomycete Huntiella omanensis
Published on: June 9, 2020
Enhancement of protoplast preparation and regeneration of Hirsutella sinensis based on process optimization
Li-Qun Jin1,2,3, Zhe-Wen Xu1,2,3, Xiao-Hui Men1,2,3
1Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, 310014, People's Republic of China.
Objective:
To explore the optimal methods for the protoplast preparation and regeneration of Hirsutella sinensis by optimizing the limiting factors.
Results:
During the treatment of enzymatic protoplast preparation, mycelium cultured for 7 days was the optimal start material. The maximum protoplast preparation rate of 4.3 × 107 protoplasts/g fresh weight (FW) was obtained after 0.5 h treatment of 1 mg/ml mixed lytic enzymes in KH2PO4-K2HPO4 buffer (pH 5.5) with 0.6 M KCl at 18 °C. As for the protoplast regeneration, the maximum protoplast regeneration rate reached 12.32% through 5 × 103 protoplasts mL-1 cultivated for 20 days in the regeneration medium with 0.6 M mannitol and 1.5% agar.
Conclusions:
The preparation and regeneration of H. sinensis protoplasts was firstly established based on process optimization and it provided a foundation for the study of H. sinensis mutagenesis.
