Fig. 6

Morphology and radial growth rates of CRISPR-HDR colonies. (A) Schematic diagram depicting the CRISPR-HDR process. The pFC332 vector, containing the Cas9 (blue), sgRNA (purple), hygromycin resistance (pink) and AMA1 (brown) components, is transformed into the fungus alongside the single strand HDR oligo. The Cas9-sgRNA complex cuts the LRG1 gene (green) at the target sequence, and the DNA is repaired using the HDR oligo to incorporate the SNP (red) into the genome. (B) Representative colony and hyphal morphology of Empty Vector, LRG1 sgRNA (gRNA), LRG1 sgRNA with Cv6 HDR oligo (gRNA + Cv6 HDR), and LRG1 sgRNA with Cv11 HDR oligo (gRNA + Cv11 HDR) transformants. Colonies were imaged after growth on PDA for seven days at 28 °C and hyphae were imaged after growth in PDB for 40 h at 28 °C. Number of colonies displaying representative morphology indicated below images. Scale bar represents 100 μm. (C) Boxplot of radial growth rate for WT, Cv6, Cv11 and selected transformant colonies averaged over a five-day period (n = 3). *** p < 0.001