Table 1. The mutagenesis frequencies of three methods for wheat genome editing in T0 generation.
| Gene | Wheat variety | Delivery form/Selection (+/−) | No. of bombarded embryos | No. of mutants/mutagenesis (%)* | No. of homozygous mutants/frequency (%)† | No. of transgene-free mutants/frequency (%)‡ | No. of homozygous transgene-free mutants/frequency (%)§ |
|---|---|---|---|---|---|---|---|
| TaGASR7 | Bobwhite | DNA/− | 1,600 | 80 (5.0) | 8 (10.0) | 35 (43.8) | 4 (5.0) |
| Kenong199 | DNA/− | 800 | 21 (2.6) | 4 (19.0) | 13 (61.9) | 1(4.8) | |
| TaGW2 | Kenong199 | DNA/+ | 800 | 24 (3.0) | 9 (37.5) | 0 | 0 |
| Kenong199 | DNA/− | 800 | 26 (3.3) | 7 (26.9) | 16 (61.5) | 2 (7.7) | |
| Kenong199 | RNA/− | 1,600 | 17 (1.1) | 6 (35.3) | 17 (100) | 6 (35.3) | |
| TaDEP1 | Kenong199 | DNA/− | 1,280 | 26 (2.0) | 0 | 14 (53.8) | 0 |
| TaNAC2 | Kenong199 | DNA/− | 800 | 16 (2.0) | N.D. | 12 (75.0) | N.D. |
| TaPIN1 | Kenong199 | DNA/− | 800 | 8 (1.0) | N.D. | 5 (62.5) | N.D. |
| TaLOX2 | Kenong199 | DNA/− | 800 | 76 (9.5) | 34 (44.7) | 66 (86.8) | 28 (36.8) |
| TdGASR7 | Shimai11 | DNA/− | 800 | 11 (1.4) | 3 (27.3) | 6 (54.5) | 0 |
| Yumai4 | DNA/− | 800 | 12 (1.5) | 5 (41.7) | 7 (58.3) | 2 (16.7) |
CRISPR, clustered regularly interspaced short palindromic repeat; N.D., not detected.
*On the basis of the number of plants carrying the observed mutations over the total number of bombarded immature embryos.
†On the basis of the number of plants carrying mutations on all the six alleles over the total number of mutant plants.
‡On the basis of the number of mutant plants not harbouring CRISPR DNA construct over the total number of mutant plants tested.
§On the basis of the number of plants carrying mutations on all the six alleles and not harbouring CRISPR DNA construct over the total number of mutant plants. ‘+' with herbicide selection; ‘−' without herbicide selection.