CRISPR/Cas9-mediated mutagenesis of phytoene desaturase in diploid and octoploid strawberry

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
Promoter	Regeneration medium ± GA₃	2 months	4 months	5 months	7 months	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
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

ISSN: 1746-4811