Fig. 2.

CLCrV-mediated targeted mutagenesis of AtGL2 and AtBRI1 in A. thaliana. a Mutant phenotype of systemically infected Cas9-OE A. thaliana leaves at 15–25 days post-infiltration with CLCrV-AtGL2-sgRNA. A. thaliana plants inoculated with CLCrV-A and CLCrV-B empty vectors served as a control. b and c Detection of AtGL2-sgRNA targeted mutations. b Col-0 served as a control, 1–7 were plant numbers. The gel image shows PCR products of the AtGL2 gene and digested PCR products with XhoI. c The undigested PCR products lacking the XhoI site (due to the presence of a mutation) that were subsequently purified, cloned, and analyzed by sequencing. d and e Detection of AtBRI1-sgRNA targeted mutations. d Col-0 served as a control, 1–7 were plant numbers. The gel image shows PCR products of the AtBRI1 gene and digested PCR products with EcoRV. e 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 in red dashes. Insertions are denoted with red capital letters