Table 2.
Transformation results with different developmentally regulated promoters driving Cre expression for auto-excision of morphogenic genes using construct design described in Figure 1 .
| Inbred | Promoter | Embryos transformed | T0 plants | T0 transformation frequency (% ± SE) | Total Single copy events | Excised single copy, backbone-free events | Excision frequency (%) | Quality event (%) | Usable events (%) |
|---|---|---|---|---|---|---|---|---|---|
| PH2RT | Ltp2 | 229 | 75 | 32.8 (2.2)a | 20 | 10 | 50.0 | 13.3 | 4.4 |
| Ole | 228 | 59 | 27.2 (3.3)ab | 20 | 8 | 40.0 | 13.6 | 3.5 | |
| Glb1 | 280 | 38 | 13.6 (1.4)c | 12 | 7 | 58.3 | 18.4 | 2.5 | |
| End2 | 174 | 39 | 22.4 (2.6)b | 3 | 3 | 100.0 | 7.7 | 1.7 | |
| Ubi | 440 | 40 | 9.1 (1.9)c | 20 | 12 | 60.0 | 30 | 2.7 | |
| HC69 | Rab17 | 121 | 35 | 28.9 (2.6)b | 4 | 1 | 25.0 | 2.9 | 0.8 |
| Ole | 151 | 49 | 37.1 (2.1)a | 8 | 3 | 37.5 | 6.1 | 2 | |
| Glb1 | 230 | 58 | 25.2 (1.8)b | 13 | 5 | 38.5 | 8.6 | 2.2 | |
| End2 | 178 | 48 | 27.0 (2.4)b | 1 | 1 | 100.0 | 2.1 | 0.6 | |
| Ubi | 202 | 37 | 18.3 (1.2)c | 22 | 3 | 13.6 | 8.1 | 1.5 |
Data presents the T0 transformation frequency, qPCR detection of the number of excised events and the quality event frequency in two different inbreds, PH2RT and HC69.
Data from three independent transformers was used to determine T0 transformation frequency. The quality events (QE) were identified as single copy, backbone-free, and morphogenic gene-free (excised). The excision frequency was determined as the ratio of the number of excised single-copy events relative to the total single-copy events. The number QEs was divided by the total number of events recovered to calculate the QE frequency. The usable event (UE) frequency is a measure of the number of acceptable transgenic events per 100 embryos that was determined as the product of QE frequency and T0 transformation frequency. Mean values followed by the same letter are not statistically different from each other at the significance level of 0.05.