Fig. 4
High frequency of genome-edited plants using CRISPR-Cas9 technology in pER8-BrrWUSa transgenic plant. a Gene-edited regeneration plants germinated from the callus. b Transgenic-specific PCR product amplified with primers Cas9-F and Cas9-R. CK, mock control; edited plants; M, DNA ladder marker DL2502. c Sequencing results of indels at the desired place. Twenty plants carried 5 different mutations including A1, A2, A5, B2, C1. B103-1 represented sequences of wild type. (d, e) Edited T0 plants showing an increased number trichomes. Asterisk (*) above the bars represents significant difference according to Student’s t-test analysis. Scale bars = 100 μm in d. f qRT-PCR analysis of BrrTCP4b targeted genes. The expression levels of all genes in the CK are set to 1. Tubulin was amplified as a constitutive control. Relative expression level of each gene was calculated by normalizing to the value in CK plants. Relative expression levels were calculated using 2−ΔΔCt method. Error bars represent Standard Deviation of triplicate experiments. Letters above the bars indicate significant difference according to one-way ANOVA analysis at P < 0.05