Editing of the ethylene biosynthesis gene in carnation using CRISPR-Cas9 ribonucleoprotein complex

Table 3 Effect of different PGR combinations and concentrations on protoplast culture

No	PGR (mg/l)	Division freq. (%)	Colony formation	Days to colony formation	No. of calli/10⁵ protoplast
1	1.0 Zeatin	−	−	−	–
2	0.5 2,4-D	63.56 ± 0.76^d	+ + +	9	−
3	1.0 2,4-D	77.12 ± 1.42^ab	+ + +	9	−
4	0.5 NAA	-	−	−	−
5	0.5 NAA 0.5 2,4-D	82.40 ± 2.10^a	+ + + +	8	984 ± 3.12^a
6	0.5 Zeatin, 1.0 2,4-D	66.83 ± 0.87^c	+ +	12	423 ± 2.11^c
7	1.0 Zeatin, 1.0 2,4-D	78.66 ± 1.24^a	+ + + +	10	1032 ± 4.13^a
8	2.0 Zeatin, 1.0 2,4-D	72.8 ± 2.33^bc	+ + +	10	765 ± 6.00^b
9	0.5 BA, 1.0 2,4-D	64.80 ± 1.52^c	+	12	272 ± 8.15^d
10	1.0 BA, 1.0 2,4-D	72.39 ± 0.91^bc	+ + +	12	474 ± 6.40^c
11	0.5 TDZ, 1.0 2,4-D	66.12 ± 0.80^c	+ +	12	551 ± 3.128^c
12	1.0 TDZ, 1.0 2,4-D	68.40 ± 1.12^c	+ + +	12	763 ± 4.77^b
13	1.0 Zeatin, 1.0 NAA	63.80 ± 3.02^d	+	10	−
14	1.0 Zeatin, 3.0 NAA	67.5 ± 0.67^c	+	10	−
15	1.0 Zeatin, 5.0 NAA	74.3 ± 1.17^b	+	10	−

*The data represent the mean of three replicates per treatment. Means with the same letter are not significantly different by Duncan’s multiple range test (DMRT, p < 0.05). Division frequency, colony formation (no. of cells that divided more than three times), and the number of calli were calculated in 8 days, 12 days, and 9 weeks after protoplast culture, respectively. —Not observed, + Poor, + + Moderate, + + + Good, + + + + Excellent

ISSN: 1746-4811