Table 3.
Detection of the on-target mutations and T-DNA in emerged plants in T1, T2, and T3 generation.
Transgenic generation | Parental genotype | Plants analyzed | Plants carrying T-DNA | Plants mutated in HvMPK6 | WT | Mutation frequency % | Biallelic Hvmpk6 mutants |
---|---|---|---|---|---|---|---|
T1 | Aa (WT/-1T) | 37 | 36 | 6 (3/3ad) | 31 | 16.22 | 0 |
T2 | A1 (WT/-1C) | 59 | 59 | 40 (38/2b) | 19 | 67.80 | 0 |
A2 (WT/-1C) | 62 | 62 | 45 (43/2b) | 17 | 72.58 | 0 | |
A3 (WT/-1C) | 24 | 24 | 17c | 7 | 70.83 | 0 | |
A4a (WT/+1T) | 43 | 43 | 17 (15/2a) | 26 | 39.53 | 0 | |
Total sequencing | A1+A2+A4 | 164 | 164 | 102 | 62 | 62.20 | 0 |
Total (WT/-1C) | A1+A2+A3 | 145 | 145 | 102 | 43 | 70.35 | 0 |
Total T2 | All | 188 | 188 | 119 | 69 | 63.30 | 0 |
T3 | A1 (WT/-1C) | 40 | 40 | 27c | 13 | 67.50 | 0 |
A2 (WT/-1C) | 20 | 20 | 13c | 7 | 65.00 | 0 | |
Total T3 | WT/-1C | 60 | 60 | 40 c | 20 | 66.67 | 0 |
Total (WT/-1C) T2+T3 | A1+A2+A3 | 205 | 205 | 142 | 63 | 69.27 | 0 |
Total T2+T3 | All | 248 | 248 | 159 | 89 | 64.11 | 0 |
In the column “Plants mutated in HvMPK6,” there are numbers in brackets. The number before slash indicates the number of heterozygotes of the WT HvMPK6 allele and mutant Hvmpk6 allele, the number after slash indicates the number of chimeras.
These plants were chimeras composed more of WT cells than of mutated cells.
These plants were multiple genotype chimeras of the WT HvMPK6 allele and two different mutated Hvmpk6 alleles.
These plants were heterozygotes of the WT HvMPK6 allele and mutant Hvmpk6 allele.
These three chimeras include also a chimeric A4 plant, which, in T1 generation, was identified as a putative heterozygous mutant in the HvMPK6 gene (WT/+1T).