Abstract
Introduction and Aim: Gnidia glauca (Fresen.) Gilg of thymelaeaceae is enlisted as an endangered plant. Hemolytic triterpenoid saponins of oleanane and ursane classes are present in G. glauca. The purpose of the present study is to increase the production of hemolytic triterpenoids in vitro using salicylic acid as an elicitor in G. glauca cell suspensions. Materials and Methods: Explants were collected and surface sterilized, then inoculated onto Murashige and Skoog medium for callus culture, further establishing suspension cultures that were treated with salicylic acid. Saponins produced in the suspension culture were extracted and analyzed by HPLC and FTIR. The hemolytic activity of saponins was confirmed by spectrophotometric assay in erythrocyte suspensions. Results: Product enhancement of hemolytic triterpenoid saponins was observed in cell suspensions treated with salicylic acid that also induced concomitant change in medium pH compared to control set of culture. Results of HPLC and FTIR analyses confirm the presence of triterpenoid saponins in suspension culture extracts. The optimal concentration of salicylic acid was found to be 200 µM for product enhancement, pH alteration. Hemolytic saponins concentration was observed to be increased with increasing concentrations of elicitor supplemented to the medium. Conclusion: Cell suspension is a viable alternative technique to produce the phytochemicals in vitro using elicitors that avoids destructive harvesting of endangered plants like G. glauca. Results of the present study demonstrate the large scale production of hemolytic saponins through cost-effective approaches.
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