Fig. 3: Detection and characterization of mutations within the VvPR4b gene in regenerated plants. | Horticulture Research

Fig. 3: Detection and characterization of mutations within the VvPR4b gene in regenerated plants.

From: CRISPR/Cas9-mediated VvPR4b editing decreases downy mildew resistance in grapevine (Vitis vinifera L.)

Fig. 3

a Identification of Hygjc2 and RTcas9 sequences in regenerated plants. Amplification in positive lines is expected to give products of 550 and 210bp, respectively. CRISPR-Cas9 plasmid DNA was used as a positive control (P). Lanes 1-14 represent distinct T0 regenerated plants tested. b Sequence of the T3 target site in regenerated plantlets. c The four distinct mutations observed in the T0 VvPR4b knockout lines. The sequence, number of regenerated clones containing the mutation, and identification number of regenerated plants are indicated. d Deduced amino acid sequences of the proteins predicted to be encoded by the mutant sequences shown in Fig. 3c. e Detection of Cas9 protein expression in the transgenic lines. Lanes 1–7 represent distinct transgenic plants. Protein extract from nontransgenic, wild-type plants (WT) was used as a negative control

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