Elicitation Approaches for Withanolide Production in Hairy Root Culture of Withania somnifera (L.) Dunal
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Withania somnifera (L.) Dunal is a versatile medicinal plant extensively utilized for production of phytochemical drug preparations. The roots and whole plants are traditionally used in Ayurveda, Unani, and Siddha medicines, as well as in homeopathy. Several studies provide evidence for an array of pharmaceutical properties due to the presence of steroidal lactones named “withanolides.” A number of research groups have focused their attention on the effects of biotic and abiotic elicitors on withanolide production using cultures of adventitious roots, cell suspensions, shoot suspensions, and hairy roots in large-scale bioreactor for producing withanolides. This chapter explains the detailed procedures for induction and establishment of hairy roots from leaf explants of W. somnifera, proliferation and multiplication of hairy root cultures, estimation of withanolide productivity upon elicitation with salicylic acid and methyl jasmonate, and quantification of major withanolides by HPLC. The protocol herein described could be implemented for large-scale cultivation of hairy root biomass to improve withanolide production.
Key wordsAgrobacterium rhizogenes Hairy root culture Salicylic acid Methyl jasmonate exposure time Inoculum mass Time course study Withanolides HPLC
Prof. A. Ganapathi is thankful to the University Grants Commissions (UGC), Government of India, for the award of UGC-BSR (Basic Scientific Research Fellowship). The first author thankfully acknowledges the Council of Scientific and Industrial Research (CSIR), Government of India, for the award of CSIR-SRF. All authors acknowledge the CSIR, Government of India, for providing financial support to carry out this work. All the authors are grateful to Dr. S. Girija of the Department of Biotechnology, Bharathiar University, Coimbatore, for providing the HPLC facility.
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