Table 2.
Summary of the target sequences and mutations of GUS, and different TaMTL and TaWaxy homologous genes in T0 wheat plants edited using the CRISPR/SpCas9-D editing system containing two sgRNAs
| Target loci | PAM-guide sequence (5´–3´) | No. of transgenic plants | No. of mutant plants | Mutation rate % | Genotypes obtained | LFD | LFD rate (%) |
|---|---|---|---|---|---|---|---|
| GUS-22 | CTTCTGCAGGTCGACTCTAGAGG | 164 | 75 | 45.7 | 24Bi + 51He + 89WT | 61 | 37.2 |
| GUS-788 | CCGTTTACGTACGTCGAGGACAT | 106 | 64.6 | 34Bi + 72He + 58WT | |||
| TaMTL4A-179 | CCAAGCTGCAGGAGCTGGACGGC | 101 | 35 | 34.7 | 11Bi + 24He + 66WT | 9 | 8.9 |
| TaMTL4A-471 | CCGCGGTGACCGCATCGCTGAGG | 36 | 35.6 | 13Bi + 23He + 65WT | |||
| TaMTL4B-179 | CCAAGCTGCAGGAGCTGGACGGG | 101 | 19 | 18.8 | 6Bi + 13He + 82WT | 6 | 5.9 |
| TaMTL4B-471 | CCGCGGTGACCGCGTCGCTGAGG | 9 | 8.9 | 7Bi + 2He + 92WT | |||
| TaMTL4D-179 | CCAAGCTGCAGGAGCTGGACGGG | 101 | 24 | 23.8 | 13Bi + 11He + 77WT | 8 | 7.9 |
| TaMTL4D-471 | CCGCGGTGACCGCGTCGCTGAGG | 16 | 15.8 | 8Bi + 8He + 85WT | |||
| TaWaxy4A-296 | GGCGGCCTCGGCGACGTCCTCGG | 87 | 35 | 40.2 | 25Bi + 10He + 52WT | 10 | 11.5 |
| TaWaxy4A-830 | AAGACCAAGGAGAAGATCTACGG | 36 | 41.4 | 29Bi + 7He + 51WT | |||
| TaWaxy7A-296 | GGCGGCCTCGGCGACGTCCTCGG | 87 | 39 | 44.8 | 19Bi + 20Bi + 48WT | 11 | 12.6 |
| TaWaxy7A-830 | AAGACCAAGGAGAAGATCTATGG | 47 | 54.0 | 25Bi + 22He + 40WT | |||
| TaWaxy7D-296 | GGCGGCCTCGGCGACGTCCTCGG | 87 | 23 | 26.4 | 10Bi + 13He + 64WT | 14 | 16.1 |
| TaWaxy7D-830 | AAGACCAAGGAGAAGATCTACGG | 31 | 35.6 | 18Bi + 13He + 56WT |
PAM, protospacer-adjacent motif, with the target sequences highlighted in bold. Bi, bi-allele; He, heterozygote; WT, wild-type; LFD, large fragment deletion.