Figure 1.

CRISPR/Cas9 Knockout of SlDDM1a and SlDDM1b Genes.

(A) Schematic illustrations of SlDDM1a and SlDDM1b gene regions targeted by the guide RNA. Black bars indicate exons and lines introns. The sgRNA-target sequences are shown, protospacer adjacent motif sequences are colored in red, black triangles mark the predicted Cas9 cut site, restriction enzyme recognition sites are underlined, and black half arrows indicate the location of primers used for mutant genotyping by PCR.

(B) Sequences of isolated mutant Slddm1a and Slddm1b alleles aligned to their respective wild types. The guide RNA target sequence is highlighted in red. PAM, protospacer adjacent motif.

(C) Scheme of SlDDM1 proteins and predicted mutant protein sequences aligned to their respective wild types below. The DNA binding domain (green), SNF2 family N-terminal domain (blue), DEAD-like helicases superfamily domain (yellow), and helicase superfamily C-terminal domain (orange) are indicated. Arrowheads indicate the site of mutation.

(D) Representative 25-d postgermination seedlings of indicated genotypes, all segregated from the same double heterozygote parent plant. Bar = 2 cm.