Figure 1.

BnCLV gene models with target sequences and schematics of binary plasmid vectors. (a) The BnCLV3 gene model includes three exons (white box) separated by two introns (represented by the solid line). The vertical line in the gene model indicates the target site, and the arrow indicates the sgRNA direction. The target sequences are shown with the PAM highlighted in red. (b) The constructs of SCLV3 and UCLV3 house the following: a hygromycin resistance cassette consisting of the hygromycin phosphotransferase coding sequence driven by the cauliflower mosaic virus 35S promoter; a Cas9 expression cassette comprising the sequence encoding Cas9 driven by P_35S or a ubiquitin promoter from maize; and two sgRNAs S1 and S2 driven by the U3b and U6‐1 promoters from Arabidopsis, respectively. (c, b) The BnCLV1 gene model with target sites S3 to S6 and the BnCLV2 gene model with target sites S7 to S10. (e) The binary constructs SCLV1 and SCLV2 with four sgRNAs driven by the U3b, U3d, U6‐1 and U6‐29 promoters from Arabidopsis. [Colour figure can be viewed at http://wileyonlinelibrary.com]