Fig. 1

Schematic diagram of the binary vector and the torenia target sequence in the torenia F3H gene. a Schematic diagram of the T-DNA region of pSKAN-pcoCas9-TfF3H used in this study. NPTII, expression cassette of NOSp-nptII-AtrbcsTer; 35Sp, CaMV35S promoter; pcoCas9, plant codon-optimized Cas9 [35]; HSPter, Arabidopsis heat shock protein 18.2 terminator [36]; RB, right border; LB, left border; AtU6p, Arabidopsis small RNA U6–26 promoter; TfF3HsgRNA, torenia F3H targeted single-guide RNA. pcoCas9 contains an intron derived from the intervening sequence 2 (IV2) of the potato St-LS1 gene [35]. b Genomic structure of the torenia F3H gene and exon 1 sequence. Boxes indicate exons, and lines between boxes indicate introns. Framed ATG indicates the start codon, and the gray box indicates the target site F3H sequence. The protospacer-adjacent motif (PAM) is underlined. Primers used for PCR amplification are also shown