Development of efficient catharanthus roseus regeneration and transformation system using agrobacterium tumefaciens and hypocotyls as explants

Table 1 Influence of transformation factors on the frequency of transient GUS expression (%) in hypocotyls of C. roseus

Factors	Variable		Transient expression rate (%)^a (mean ± SE)^c	Death rate (%)^a (mean ± SE)^b
A. tumefaciens infection density(OD) and duration (min)	0.5	15	53.33 ± 3.33 c	10.00 ± 2.89 b
		30	78.33 ± 4.41 ab	11.67 ± 1.67 a
		45	80.00 ± 2.89 a	11.67 ± 3.33 a
	0.8	15	78.33 ± 3.33 b	15.00 ± 1.67 b
		30	100.00 ± 0.00 a	15.00 ± 2.89 b
		45	100.00 ± 0.00 a	21.67 ± 1.67 a
	1.0	15	73.33 ± 1.67 b	18.33 ± 3.33 c
		30	100.00 ± 0.00 a	46.66 ± 6.01 b
		45	100.00 ± 0.00 a	68.33 ± 4.41 a
Co-cultivation period (d)	1		76.67 ± 4.41 b	0.00 ± 0.00 c
	2		100.00 ± 0.00 a	5.00 ± 2.89 b
	3		100.00 ± 0.00 a	20.00 ± 5.77 a
Acetosyringone concentration (μM)	0		13.33 ± 3.33 c	0.00 ± 0.00 cd
	50		85.00 ± 2.89 ab	1.66 ± 1.66 bc
	100		100.00 ± 0.00 a	5.00 ± 2.89 b
	150		100.00 ± 0.00 a	46.67 ± 4.41 a

Transient expression of GUS gene was examined after co-culture. The data of death rates were collected after 10 d of co-culture. Hypocotyls were placed on MSCP1 medium.
^a Transient expression rate = number of GUS⁺(GUS-positive) explants/number of explants × 100.
^b Death rate = number of died explants/number of explants × 100.
^c Values represent means for 60 explants per treatment, replicated three times. Mean ± SE in each column followed by the same letter are not significantly different according to Tukey’s multiple comparison test (P ≤ 0.05).

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