Figure 1 The principle for microhomology-mediated end joining (MMEJ)-based DNA fragment deletion(modified from Xie et al., 2020) MHS: Microhomologous sequence

Figure 2 The experimental workflow for microhomology-mediated end joining (MMEJ)-mediated DNA fragment deletion

Figure 3 The strategy for identification of the mutant types in transgenic plants (A) Amplification of the target region using the forward primer (T1-F) upstream the T1 and reverse primer (T2-R) downstream T2 if the length of desirable deleted fragment is less than 800 bp; (B) Four primers (with the length of the following amplification products is about 70-100 bp different from each other) with three combinations (T1-F/T1-R, T2-F/T2-R, and T1-F/T2-R) or pooled in an amplification are suggested if the length of desirable deleted fragment is more than 800 bp (there is no deletion of the target region if the resulting bands are produced with primer groups (T1-F/T1-R and T2-F/T2-R) but not T1-F/T2-R; both alleles had fragment deletion if the resulting bands are produced with primer group T1-F/T2-R but not T1-F/T1-R and T2-F/T2-R; one allele had fragment deletion and the other allele had no fragment deletion if all three fragments were produced; (C) The example of gel electrophoresis according to the detection method of (A). WT: Wild type; 1-9 indicate edited T0 plants in which 1-4 with biallelic fragment deletion, 5, 6, 8 and 9 with heterozygous deletion and 7 with no deletion.