Table 1 Transformation and regeneration statistics for eight constructs using Agrobacterium strain EHA105 to transform CO39 rice calli derived from mature seeds, utilizing a modified transformation protocol; including additional l-Proline (round 1–5), removal of antibiotics from regeneration media (rounds 2–5) and transformation of calli 8 days after induction instead of 18 days (4–5)
Transformation round | Candidate gene | Construct size LB-RB (kb) | Number of Agrobacterium infected calli | Independent proliferating calli lineages after selection | Putative transformation efficiency (%) | Regenerated plantlets (counting maximum 1 per event/lineage) | Putative regeneration efficiency (%) | Escapes killed by antibiotic selection on rooting media | Escapes (surviving until genotyping) | Independent transformation events after genotyping | Overall efficiency (%) |
|---|---|---|---|---|---|---|---|---|---|---|---|
1 | LOC_Os08g10440 | 11.4 | 97 | 59 | 60.82474 | 2 | 3.389831 | 0 | 2 | 2.06 | |
LOC_Os12g18729 | 13.6 | 104 | 29 | 27.88462 | 2 | 6.896552 | 0 | 2 | 1.92 | ||
2 | LOC_Os12g22839 | 6.6 | 180 | 46 | 25.55556 | 9 | 19.56522 | 4 | 5 | 2.78 | |
LOC_Os12g22880 | 8.3 | 180 | 47 | 26.11111 | 12 | 25.53191 | 3 | 1 | 8 | 4.44 | |
3 | LOC_Os08g10440 | 11.4 | 201 | 73 | 36.31841 | 13 | 17.80822 | 2 | 11 | 5.47 | |
LOC_Os12g18729 | 13.6 | 238 | 74 | 31.09244 | 24 | 32.43243 | 6 | 1 | 17 | 7.14 | |
4 | LOC_Os09g12350 | 14.5 | 340 | 64 | 18.82353 | 24 | 37.5 | 11 | 1 | 12 | 3.53 |
LOC_Os09g12380 | 9 | 380 | 85 | 22.36842 | 35 | 41.17647 | 22 | 13 | 3.42 | ||
5 | LOC_Os09g17810 | 10.3 | 100 | 23 | 23 | 9 | 39.13043 | 2 | 7 | 7.00 | |
LOC_Os09g17870 | 9.2 | 100 | 53 | 53 | 16 | 30.18868 | 4 | 1 | 11 | 11.00 |