Figure 6.

TaNOX10 plays a positive role in ROS production and wheat resistance to Pst. A, Schematic diagram of TaNOX10 gene structure and the sequences of the two sgRNAs designed to target the three homoeoalleles (A, B, D), of TaNOX10 for editing by CRISPR–Cas9. Black rectangle, exon. Blue letters, PAM. B, Sequences of the WT TaNOX10 and tanox10-knockout (tanox10-ko) plant at the site targeted by sgRNA. The tanox10-ko line contains frameshift mutations in tanox10-A (1-bp insertion), tanox10-B (5-bp deletion), and tanox10-D (1-bp insertion). C, tanox10-ko and the WT KN199 variety were inoculated with Pst race CYR32, and their disease phenotypes were observed at 14 dpi. D, Extracellular H₂O₂ contents in tanox10-ko and WT plants at 24 hpi with Pst CYR32. Data are shown as means ± SD from three biological replicates. Statistical significance was determined by Student’s t test. *P < 0.05. E, Fungal biomass in tanox10-ko and WT at 14 dpi with Pst CYR32, as estimated by qPCR of PsEF1 and wheat TaEF1α DNAs in infected samples and calculated with reference to gene-specific standard curves. Data are shown as means ± SD from three biological replicates. Statistical significance was determined by Student’s t test. **P < 0.01.