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<p><i>Withania somnifera</i>, commonly known as Ashwagandha, is a herbal remedy that has been used in Ayurvedic and traditional medicine for over 3,000 years. It is known for its adaptogenic benefits, as its major metabolite, Withaferin A, contributes to these effects. This study focuses on various biotic and abiotic elicitation techniques that significantly improve biomass and secondary metabolite production in in vitro shoots, outperforming previous studies on single elicitors. The shoot cultures were cultivated on Murashige and Skoog medium enriched with 4.44 µM benzylaminopurine in suspension cultures and subjected to elicitation treatments using seaweed extract (<i>Kappaphycus alvarezii</i>) at concentrations of 0.15%, 0.25%, and 0.50% (w/v), silver nitrate at 0.5, 1.0, and 2.0 mM, and lead acetate at 0.2, 0.4, and 0.8 mM for 7 and 14 days’ duration. The responses were evaluated by calculating growth index, and Withaferin A content was measured using ultra-high-performance liquid chromatography coupled with tandem mass spectrometry. In untreated in vitro control shoots, the Withaferin A content was found to be 0.818&#xa0;mg/g. The highest accumulation of Withaferin A occurred with 0.25% w/v seaweed extract (2.777&#xa0;mg/g) for 7 days, followed by 1.0 mM silver nitrate (1.156&#xa0;mg/g) and 0.8 mM lead acetate (1.400&#xa0;mg/g) for 14 days duration. These findings underscore the effectiveness of elicitor-based strategies in promoting secondary metabolite production under controlled in vitro conditions and the robustness of UHPLC-MS/MS for accurate metabolite profiling in plant tissue cultures.</p>

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Elicitor- mediated enhancement of Withaferin A in vitro shoots of Withania somnifera – a quantitative study using UHPLC-MS/MS

  • 期刊论文

  • 发表日期：

    2026年1月31日

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  • Aparnapreethi Rajendran,
  • Kanimozhi Natarajan,
  • Karthikeyan Prakasham,
  • Vellaikumar Sampathrajan,
  • Kalaiselvi Senthil

摘要

Withania somnifera, commonly known as Ashwagandha, is a herbal remedy that has been used in Ayurvedic and traditional medicine for over 3,000 years. It is known for its adaptogenic benefits, as its major metabolite, Withaferin A, contributes to these effects. This study focuses on various biotic and abiotic elicitation techniques that significantly improve biomass and secondary metabolite production in in vitro shoots, outperforming previous studies on single elicitors. The shoot cultures were cultivated on Murashige and Skoog medium enriched with 4.44 µM benzylaminopurine in suspension cultures and subjected to elicitation treatments using seaweed extract (Kappaphycus alvarezii) at concentrations of 0.15%, 0.25%, and 0.50% (w/v), silver nitrate at 0.5, 1.0, and 2.0 mM, and lead acetate at 0.2, 0.4, and 0.8 mM for 7 and 14 days’ duration. The responses were evaluated by calculating growth index, and Withaferin A content was measured using ultra-high-performance liquid chromatography coupled with tandem mass spectrometry. In untreated in vitro control shoots, the Withaferin A content was found to be 0.818 mg/g. The highest accumulation of Withaferin A occurred with 0.25% w/v seaweed extract (2.777 mg/g) for 7 days, followed by 1.0 mM silver nitrate (1.156 mg/g) and 0.8 mM lead acetate (1.400 mg/g) for 14 days duration. These findings underscore the effectiveness of elicitor-based strategies in promoting secondary metabolite production under controlled in vitro conditions and the robustness of UHPLC-MS/MS for accurate metabolite profiling in plant tissue cultures.