Fig. 5

Co-expression of p19 and CRISPR/Cas9 in Arabidopsis transgenic plants. a Schematic diagram of the p19 co-expression CRISPR/Cas9 system, pIUC-p19. p19, tomato bushy stunt virus p19 gene; 2A, Porcine teschovirus-1 2A peptide. b T1 transformants exhibited leaf developmental phenotypes with different severities. “I”: WT looking plants; “II”: mild phenotype with curly leaves; “III”: severe phenotype with serrated leaves. c p19 protein levels in T1 transgenic lines exhibiting different leaf phenotype severities were detected by Western Blot using anti-2A antibodies. d Histogram showing the number of WT, chimeras and mutant plants in T1 populations transformed with pIUC and pIUC-p19. For each system, two different genes, TT4 and AP1, were targeted. e Re-analysis of the data shown in (d) grouping plants according to the severity of their leaf phenotype. The numbers on the top of each column indicate the proportion of grouped phenotypes in the population f Cas9 transcript levels in T1 transgenic lines exhibiting different leaf phenotype severities determined by quantitative RT-PCR. The columns indicate mean ± SD of three biological replicates. Statistic was performed with t test in comparison to pIUC-1. *Significant difference at p < 0.05 under two-tailed Student’s t test. **Significant difference at p < 0.01 under two-tailed Student’s t test. “p > 0.05” indicates no significant difference. g, sgRNA and miR168 expression levels in T1 transgenic lines exhibiting different leaf phenotype severities detected by Northern blot analysis