Fig. 3.

Detection of mutations in T0 DA1-editing plants using a PCR-restriction enzyme (PCR-RE) assay. a Schematic map of the binary vector of the DA1 gene for genome editing in wheat. Cas9 is expressed with a ubiquitin promoter. The synthetic guide RNA (sgRNA) is derived using U3 promoters. b Gel electrophoresis of PCR products amplified from the mutated region of the A genome with specific primers and digested with SphI. Lanes 1–21 are the digested DNA of the PCR products amplified from different transgenic plants; lane 22 is a wild-type sample. c Gel electrophoresis of PCR products amplified from the mutated region of the B genome with specific primers and digested with SphI. Lanes 1–23 are the digested DNA of the PCR products amplified from different transgenic plants; lane 24 is a wild-type sample. d Gel electrophoresis of PCR products amplified from the mutated region of the D genome with specific primers and digested with SphI. Lanes 1–23 are the digested DNA of the PCR products amplified from different transgenic plants; lane 24 is a wild-type sample