Figure 1.
Growth kinetics and withanolides production in cell suspension culture of W. somnifera.
a. Dynamic profiles of biomass accumulation, b. Dynamic profiles of withanolides production, c. Synchronized cell suspension culture in 150-flask culture, d. Lab scale cell suspension culture in 7-l bioreactor. The cell suspension cultures were established by inoculating ∼500 mg fresh mass of friable callus in 150 ml Erlenmeyer flask containing 30 ml of MS liquid medium supplemented with 1 mg/l picloram, 0.5 mg/l KN, 200 mg/l L-glutamine and 5% sucrose and kept on gyratory shaker at 120 rpm under total darkness. Bioreactor culture was established with 83 g FW of friable callus in 5-l MS liquid medium with same hormonal combinations. Values represent mean ±standard error of three replicates; each experiment was repeated thrice. All bars = 1 cm.
Table 1.
Productivity of withanolides in shake-flask culture and bioreactor by elicitor and precursor treatments in cell suspension of W. somnifera.
Figure 2.
The effect of different concentrations of chitosan on growth characteristics (a) and withanolides production (b) in cell suspension culture of W. somnifera in shake-flask culture at 4 h exposure time.
Five hundred milligram of fresh mass of friable callus was cultured in 30/l picloram, 0.5 mg/l KN, 200 mg/l L-glutamine and 5% sucrose and kept on gyratory shaker at 120 rpm under total darkness. The cultures were harvested on 28th day. Data represents mean±standard error of three replicates; each experiment was repeated thrice.
Figure 3.
The effect of different concentrations of aluminium chloride on growth characteristics (a) and withanolides production (b) in cell suspension culture of W. somnifera in shake-flask culture at 4 h exposure time.
Five hundred milligram of fresh mass of friable callus was cultured in 30/l picloram, 0.5 mg/l KN, 200 mg/l L-glutamine and 5% sucrose and kept on gyratory shaker at 120 rpm under total darkness. The cultures were harvested on 28th day. Data represents mean±standard error of three replicates; each experiment was repeated thrice.
Figure 4.
The effect of different concentrations of cadmium chloride on growth characteristics (a) and withanolides production (b) in cell suspension culture of W. somnifera in shake-flask culture at 4 h exposure time.
Five hundred milligram of fresh mass of friable callus was cultured in 30/l picloram, 0.5 mg/l KN, 200 mg/l L-glutamine and 5% sucrose and kept on gyratory shaker at 120 rpm under total darkness. The cultures were harvested on 28th day. Data represents mean±standard error of three replicates; each experiment was repeated thrice.
Figure 5.
The effect of different concentrations of squalene on biomass accumulation (a) and withanolides production (b) in cell suspension culture of W. somnifera in shake-flask culture at 48 h exposure time.
Five hundred milligram of fresh mass of friable callus was cultured in 30/l picloram, 0.5 mg/l KN, 200 mg/l L-glutamine and 5% sucrose and kept on gyratory shaker at 120 rpm under total darkness. The cultures were harvested on 28th day. Data represents mean±standard error of three replicates; each experiment was repeated thrice.
Figure 6.
The effect of different concentrations of mevalonic acid on biomass accumulation (a) and withanolides production (b) in cell suspension culture of W. somnifera in shake-flask culture at 48 h exposure time.
Five hundred milligram of fresh mass of friable callus was cultured in 30/l picloram, 0.5 mg/l KN, 200 mg/l L-glutamine and 5% sucrose and kept on gyratory shaker at 120 rpm under total darkness. The cultures were harvested on 28th day. Data represents mean±standard error of three replicates; each experiment was repeated thrice.
Figure 7.
The effect of different concentrations of cholesterol on biomass accumulation (a) and withanolides production (b) in cell suspension culture of W. somnifera in shake-flask culture at 48 h exposure time.
Five hundred milligram of fresh mass of friable callus was cultured in 30/l picloram, 0.5 mg/l KN, 200 mg/l L-glutamine and 5% sucrose and kept on gyratory shaker at 120 rpm under total darkness. The cultures were harvested on 28th day. Data represents mean±standard error of three replicates; each experiment was repeated thrice.