Effect of Salts (NaCl and Na2CO 3) on Callus and Suspension Culture of Stevia rebaudiana for Steviol glycoside Production
2014
Gupta, Pratibha | Sharma, Satyawati | Saxena, Sanjay
Steviol glycosides are natural non-caloric sweeteners which are extracted from the leaves of Stevia rebaudiana plant. Present study deals the effect of salts (NaCl and Na₂CO₃) on callus and suspension culture of Stevia plant for steviol glycoside (SGs) production. Yellow-green and compact calli obtained from in vitro raised Stevia leaves sub-cultured on MS medium supplemented with 2.0 mg l⁻¹NAA and different concentrations of NaCl (0.05–0.20 %) and Na₂CO₃(0.0125–0.10 %) for 2 weeks, and incubated at 24 ± 1 °C and 22.4 μmol m⁻² s⁻¹light intensity provided by white fluorescent tubes for 16 h. Callus and suspension biomass cultured on salts showed less growth as well as browning of medium when compared with control. Quantification of SGs content in callus culture (collected on 15th day) and suspension cultures (collected at 10th and 15th days) treated with and without salts were analyzed by HPLC. It was found that abiotic stress induced by the salts increased the concentration of SGs significantly. In callus, the quantity of SGs got increased from 0.27 (control) to 1.43 and 1.57 % with 0.10 % NaCl, and 0.025 % Na₂CO₃, respectively. However, in case of suspension culture, the same concentrations of NaCl and Na₂CO₃enhanced the SGs content from 1.36 (control) to 2.61 and 5.14 %, respectively, on the 10th day.
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