Identification of the 20-nt target recognition sequences and editing efficiency evaluation of the constructed transgenic collection in the T0 generation. (A) The ratio of the correct target recognition sequences to the mutated ones. (del.) Deletion, (ins.) insertion, and (sub.) substitution. (B) Editing efficiency analysis with 86 randomly selected positive transgenic plants. (WT) Wild type. (C) Editing evaluation of the 178 genes in the 86 selected transgenic lines. (D) The proportion of the number of insertions (n = 79) to that of the deletions (n = 47). (E) Percentages of different lengths of inserted and deleted base pairs. (F) Evaluation of editing efficiency of sgRNAs targeting different numbers of homoeologs. The numbers inside the polar plot represent the ratios of mutagenic sgRNAs to the total examined that target the same number of homoeologs; the ones alongside indicate the numbers of homoeologs. The top three bar plots detail the percentages of sgRNAs editing different numbers of homoeologs. For sgRNAs targeting one to four homoeologs: n = 40, n = 41, n = 4, n = 11. (G) Editing identification of the six transgenics harboring two distinct sgRNAs and two transgenic plants with three different sgRNAs. The eight transgenic plants were randomly selected for the examination.
