Figure 3.

CRISPR/Cas9-mediated S-RNase mutations result in self-compatible N. alata. (A) S-RNase allele identification in N. alata. Primer sets S2F/R and Sc10F/R were used for PCR analysis to determine the presence of S2 and Sc10 alleles in the population used for gene editing. (B) Cas9-PF was armed with a polycistronic tRNA-gRNA (PTG) cassette consisting of S2 and Sc10 gRNAs. (C) Mutation patterns in T0 transgenic plants. sgRNA, single guide RNA; PAM, the protospacer adjacent motif; WT, wild type; inserts and deletions are indicated as ‘i’ and ‘d’ respectively. (D) compared to the sterility of WT, well-stacked fruits and mounts of seeds were harvested in two bi-allele edited N. alata plants. Scale bar=1 cm. (E) Segregation ratio of Cas9-PF-free plants and S2 and Sc10 alleles in T1 generation of two bi-allele edited N. alata lines respectively.