Enhanced production of silymarin by Ag⁺ elicitor in cell suspension cultures of Silybum marianum

Figures & data

Figure 1.  Chemical structure of silybin A (1), silybin B (2), isosilybin A (3), isosilybin B (4), silychristin (5), and silydianin (6).

Figure 2.  Time courses of S. marianum cell growth (▪) and silymarin production (□) in cell suspension culture. 10 mL suspension culture containing about 1 g Fresh Weight cells was incubated in 40 mL medium in a 100 mL flask on a rotary shaker (120 rpm). Data are the average of three experiments each in triplicate (mean ± SD).

Figure 3.  Effects of feeding time in S. marianum cell cultures. Three-day-old cultures were treated with Ag⁺ (A 0.2, B 0.4, C 0.8, D 1, and E 2 mM). Cells were analyzed for dry weight at the indicated times; (▪) treated cultures, (□) control cultures. Data are the average of three experiments each in triplicate (mean ± SD).

Figure 4.  Effects of feeding time in S. marianum cell cultures. Three-day-old cultures were treated with Ag⁺ (A 0.2, B 0.4, C 0.8, D 1, and E 2 mM). Cells were analyzed for silymarin production at the indicated times; (•) treated cultures, (▴) control cultures. Data are the average of three experiments each in triplicate (mean ± SD).

Table 1.  Effects of feeding time in S. marianum cell cultures.

Incubation period (h)	Ag^+ concentration (mM)	TXF (µg g^−1 DW)	SC (µg g^−1 DW)	SD (µg g^−1 DW)	SB (µg g^−1 DW)	ISB (µg g^−1 DW)	SLM (µg g^−1 DW)
12	0	0.034 ± 0.06	1.42 ± 0.92	2.51 ± 0.61	0.41 ± 0.18	0.34 ± 0.14	4.72 ± 1.018
	0.2	3.25 ± 0.1	21.2 ± 0.99	6.02 ± 1.93	4.9 ± 0.13	2.15 ± 1.05	37.52 ± 1.8
	0.4	1.26 ± 0.3	10.52 ± 0.96	3.85 ± 0.97	2.3 ± 0.98	1.02 ± 0.3	18.95 ± 2
	0.8	0.71 ± 0.06	4.52 ± 0.98	1.4 ± 0.54	0.85 ± 0.18	0.16 ± 0.012	7.64 ± 1.73
	1	0.3 ± 0.004	3.51 ± 1.2	0.82 ± 0.18	0.51 ± 0.19	0.25 ± 0.013	5.39 ± 1.4
	2	0.043 ± 0.01	4.9 ± 0.9	1.65 ± 0.19	1.86 ± 1.03	0.6 ± 0.13	9.053 ± 1.9
24	0	0.005 ± 0.002	1.82 ± 0.62	0.56 ± 0.206	0.23 ± 0.032	0.23 ± 0.04	2.85 ± 0.72
	0.2	3.4 ± 0.5	24.1 ± 1.7	7.32 ± 1.24	11 ± 1.51	2.84 ± 0.7	48.66 ± 1.7
	0.4	4.36 ± 0.91	28.8 ± 1.81	5.4 ± 1.05	4.34 ± 1.12	1.72 ± 0.51	44.62 ± 2.2
	0.8	14.95 ± 0.83	27.5 ± 1.22	6.13 ± 1.47	6.55 ± 1.8	1.12 ± 0.6	56.25 ± 1.35
	1	1.82 ± 0.74	9.35 ± 1.01	2.96 ± 1.3	1.91 ± 1.6	2 ± 0.4	18.04 ± 1.24
	2	1.68 ± 0.25	5.15 ± 0.49	1.54 ± 0.92	2.94 ± 1.0	2.04 ± 0.52	13.36 ± 1.5
48	0	0.35 ± 0.05	0.76 ± 0.2	0.6 ± 0.066	0.8 ± 0.74	0.3 ± 0.07	2.85 ± 0.24
	0.2	5.92 ± 0.86	15.93 ± 1.78	2.75 ± 1.03	3.23 ± 0.61	1.52 ± 0.39	29.37 ± 1.84
	0.4	3.55 ± 0.36	16.71 ± 1.06	4.72 ± 1.07	3.21 ± 1.53	0.46 ± 0.02	28.65 ± 2.33
	0.8	4.09 ± 0.04	19.83 ± 1.05	6.48 ± 1.97	3.33 ± 1.32	1.4 ± 0.34	35.13 ± 2.4
	1	1.74 ± 0.21	4.71 ± 0.7	3.23 ± 0.7	2.63 ± 1.63	1.37 ± 0.06	13.7 ± 1.32
	2	3.25 ± 0.13	11.34 ± 0.76	4.44 ± 1.21	3.07 ± 1.03	1.58 ± 0.08	23.7 ± 1.83
72	0	0.012 ± 0.002	1.33 ± 0.11	1.1 ± 0.27	0.2 ± 0.05	0.23 ± 0.004	2.98 ± 0.57
	0.2	3.5 ± 0.03	15.17 ± 1.77	4.3 ± 1.02	3.43 ± 1.35	1.02 ± 0.01	27.42 ± 1.55
	0.4	2.38 ± 0.14	14 ± 1.70	3.71 ± 1.08	2.14 ± 0.92	0.55 ± 0.005	22.8 ± 1.4
	0.8	6.21 ± 0.71	15.22 ± 1.34	5.43 ± 0.74	4 ± 1.39	1.34 ± 0.6	32.2 ± 1.24
	1	2.5 ± 0.5	6 ± 1.21	2.5 ± 0.51	2.74 ± 1.04	0.7 ± 0.0051	14.44 ± 1.92
	2	1.85 ± 0.05	4 ± 0.8	2.45 ± 0.84	0.87 ± 0.3	0.45 ± 0.061	8.54 ± 1.5
144	0	0.03 ± 0.001	0.8 ± 0.078	0.7 ± 0.4	0.17 ± 0.003	0.26 ± 0.001	1.93 ± 0.34
	0.2	3.85 ± 0.15	14.73 ± 1.16	4.9 ± 1.05	2.4 ± 0.29	0.62 ± 0.005	26.5 ± 1.51
	0.4	1.17 ± 0.72	6.24 ± 0.92	2.76 ± 0.16	1.11 ± 1.75	0.5 ± 0.03	11.8 ± 1.32
	0.8	2.92 ± 0.52	17.66 ± 1.102	2.41 ± 0.42	2.51 ± 0.94	0.99 ± 0.12	26.5 ± 2
	1	0.8 ± 0.22	3.56 ± 1.28	2 ± 1.25	1.36 ± 1.35	0.48 ± 0.01	8.2 ± 1.22
	2	1.011 ± 0.6	4.8 ± 1.45	3.1 ± 1.24	1.06 ± 1.16	0.24 ± 0.007	10.21 ± 1.87
216	0	0.955 ± 0.085	1.4 ± o.32	1.1 ± 0.74	0.13 ± 0.025	0.19 ± 0.006	3.78 ± 0.56
	0.2	2.6 ± 0.12	13.76 ± 1.49	3.09 ± 1.12	3.4 ± 0.92	1.3 ± 0.2	24.15 ± 1.05
	0.4	6.18 ± 0.71	17.108 ± 1.93	4.47 ± 1.67	4.14 ± 1.65	0.54 ± 0.001	32.45 ± 1.3
	0.8	1.6 ± 0.11	7 ± 1.67	3.1 ± 1.32	1.91 ± 1.08	0.81 ± 0.07	14.42 ± 1.44
	1	11.91 ± 0.64	30.6 ± 1.27	3.09 ± 1.001	9.5 ± 0.77	4.4 ± 0.98	59.5 ± 1.06
	2	3.22 ± 0.6	15.51 ± 1.05	3.1 ± 0.81	2.65 ± 1.05	0.92 ± 0.03	25.4 ± 1.93

Note. Three-day-old cultures were treated with Ag⁺ (0.2, 0.4, 0.8, 1, and 2 mM). Cells were analyzed for silybin (SB), isosilybin (ISB), taxifolin (TXF), silychristin (SC), and silydianin (SD) production at the indicated times; data are the average of three experiments each in triplicate (mean ± SD).