Table 1.
Numbers and percentages of ‘Hawaii 4’ and ‘Calypso’ shoot lines regenerating on selective medium
Promoter | Regeneration medium ± GA3 | Number of lines harvested throughout the experiment | Total lines harvested | Number of lines assessed for transformed phenotype | Putative transformed lines | % Putative transformed lines | Lines with mutant phenotype | % Putative transformants with mutant phenotype | Cultivar | |||
---|---|---|---|---|---|---|---|---|---|---|---|---|
2 months | 4 months | 5 months | 7 months | |||||||||
At | + GA | 18 | 18 | 22 | 7 | 65 | 58 | 42 | 72 | 24 | 57 | Hawaii-4 |
At | no GA | 1 | 4 | 1 | 0 | 6 | 6 | 5 | 83 | 1 | 20 | |
Fv | + GA | 12 | 1 | 39 | 4 | 56 | 55 | 50 | 91 | 46 | 92 | |
Fv | no GA | 15 | 8 | 15 | 0 | 38 | 37 | 26 | 70 | 21 | 81 | |
Average | 79 | 62 | ||||||||||
At | + GA | 0 | 11 | 10 | 21 | 42 | 30 | 22 | 73 | 8 | 36 | Calypso |
At | no GA | 0 | 9 | 7 | 6 | 22 | 19 | 16 | 84 | 11 | 69 | |
Fv | + GA | 2 | 4 | 1 | 6 | 13 | 9 | 8 | 89 | 4 | 50 | |
Fv | no GA | 2 | 3 | 4 | 3 | 12 | 10 | 8 | 80 | 2 | 25 | |
Average | 82 | 45 |
Explants were regenerated on selective medium with (+) or without (no) GA3, following transformation with sgRNAs under transcriptional control of the Arabidopsis (At) or the F. vesca (Fv) U6III promoters. ‘Numbers of lines harvested throughout the experiment’ are callus lines initiating shoots at 2, 4, 5 and 7 months. ‘Putative transformed lines’ are lines either forming roots on medium supplemented with kanamycin and/or regenerating shoots with a mutant phenotype