Journal: Journal of Fungi
Article Title: Unlocking the Potential for Genetic Engineering of the Straw-Degrading Mushroom Stropharia rugosoannulata by Constructing a CRISPR/Cas9 Gene Editing System
doi: 10.3390/jof12040269
Figure Lengend Snippet: Establishment of an ATMT system for S. rugosoannulata mycelium. ( A ) The GPiE-SrU6 plasmid carrying two knockout targets was introduced into A. tumefaciens GV3101 for amplification, followed by cocultivation of wounded mycelium with Agrobacterium for 96 h. ( B ) After cocultivation, the mycelium was washed in medium containing 600 mg/L Cef, inoculated onto PDA medium supplemented with 60 µg/mL Hyg for primary screening, and subsequently transferred onto PDA medium containing 0.2 mg/mL 5-FOA for secondary screening. Growth on different selective drugs (Cef, Hyg and 5-FOA), with three biological replicates for each treatment. ( C ) Gel electrophoresis image of the hygromycin gene amplified from DNA extracted from some candidate transgenic strains.
Article Snippet: Furthermore, S. rugosoannulata was inoculated onto PDA resistance medium supplemented with 0, 0.05, 0.1, 0.2, or 0.3 mg/mL 5-FOA (Solarbio Science & Technology Co., Ltd., Beijing, China).
Techniques: Plasmid Preparation, Knock-Out, Amplification, Nucleic Acid Electrophoresis, Transgenic Assay