Fig. 3

FT-sgRNA targeted mutagenesis of AtBRI1 in A. thaliana. a Truncated FT RNA was fused to the 5′ end of AtBRI1-sgRNA. b and c Detection of FT-AtBRI1-sgRNA targeted mutations in Pro35S::Cas9 and ProYao::Cas9 A. thaliana. Col-0 served as a control, 1–12 were plant numbers. The gel image shows digested PCR products of the AtBRI1 gene with EcoRV, the undigested PCR products lacking the EcoRV site (due to the presence of a mutation) that were subsequently purified, cloned, and analyzed by sequencing. The green color indicated the PAM sequence. The restriction site on the target sequence is underlined in blue. M indicates the mutation sequence. Deletions are shown with red dashes. Insertions are denoted with red capital letters. Substitutions are denoted with red lowercase letters. d Phenotype of bri1 mutations in M2 generation. The red circle shows the bri1 mutant