Fig. 1.

Generation of BnaPPT1 overexpression transgenic plants and knockout mutants. (a) Amino acid sequence alignment using MEGA7 and gene expression of PPTs in B. napus. At, Arabidopsis thaliana; Bn, B. napus; Gene expression data are from BnTIR (http://yanglab.hzau.edu.cn/). (b) BnaC08.PPT1 is a chloroplast membrane protein observed in Arabidopsis protoplast. Bars = 10 μm. (c) Relative expression level of BnaC08.PPT1 in overexpression lines by quantitative real-time PCR. Total RNA was extracted from 8-week-old leaves. Values are means ± SD (n = 3). ** indicates P < 0.01, based on a student t-test. (d) CRSIPR targets on BnaA08.PPT1 and BnaC08.PPT1. Two target sites are designed on the first and third exons respectively. (e) Acquisition of mutant lines by sequencing. CR5 and CR105 is a homozygous mutant line of BnaA08.PPT1 and BnaC08.PPT1 respectively. CR28 is a homozygous double mutant, and CR54 is a heterozygous double mutant. Mutation sites are indicated in red, and PAM sites are indicated in blue. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)