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<p><i>Leucojum aestivum</i> L. (Amaryllidaceae) is a valuable medicinal bulbous plant and a natural source of galanthamine, an alkaloid used in Alzheimer’s disease treatment. However, its low natural yield and overharvesting have threatened its wild populations, emphasizing the need for sustainable production strategies. This study aimed to optimize in vitro bulblet proliferation and galanthamine accumulation through hormonal elicitation using ethephon (an ethylene-releasing compound) in combination with auxins (NAA and IBA) on Murashige and Skoog (MS) medium. Among the treatments, 1 mg/L ethephon with 1.5 mg/L IBA significantly enhanced bulblet number (6.00 ± 0.58, a six-fold increase over control) and galanthamine content (2.024 ± 0.097 mg/g DW, ten-fold higher than control), as determined by HPLC. Antioxidant responses to elicitation stress included marked increases in enzymatic activities superoxide dismutase (SOD), catalase (CAT), and phenylalanine ammonia-lyase (PAL) with SOD activity increasing up to 7.5-fold under 1 mg/L ethephon + 1 mg/L NAA. Non-enzymatic antioxidants, including total phenolics and FRAP values, were also elevated, indicating improved redox homeostasis. Correlation analysis showed strong positive relationships (<i>r</i> &gt; 0.85) between galanthamine levels, bulblet proliferation, and antioxidant capacity, suggesting coordinated metabolic and morphogenic responses. Interestingly, enzymatic antioxidants exhibited inverse correlations with alkaloid content. These findings demonstrate the synergistic effect of ethephon and IBA in enhancing both biomass and secondary metabolite production. The optimized protocol provides an efficient and sustainable platform for in vitro propagation and high-value alkaloid biosynthesis in <i>L. aestivum</i>, contributing to both conservation and pharmaceutical applications.</p>

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Synergistic effect of ethephon and auxin on in vitro bulblet proliferation and galanthamine biosynthesis in summer snowflake (Leucojum aestivum L.)

  • 期刊论文

  • 发表日期：

    2025年12月22日

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  • Maryam Mollaahmadi,
  • Hassan Esmaeili,
  • Mahdi Moridi Farimani,
  • Mohammad Hossein Mirjalili

摘要

Leucojum aestivum L. (Amaryllidaceae) is a valuable medicinal bulbous plant and a natural source of galanthamine, an alkaloid used in Alzheimer’s disease treatment. However, its low natural yield and overharvesting have threatened its wild populations, emphasizing the need for sustainable production strategies. This study aimed to optimize in vitro bulblet proliferation and galanthamine accumulation through hormonal elicitation using ethephon (an ethylene-releasing compound) in combination with auxins (NAA and IBA) on Murashige and Skoog (MS) medium. Among the treatments, 1 mg/L ethephon with 1.5 mg/L IBA significantly enhanced bulblet number (6.00 ± 0.58, a six-fold increase over control) and galanthamine content (2.024 ± 0.097 mg/g DW, ten-fold higher than control), as determined by HPLC. Antioxidant responses to elicitation stress included marked increases in enzymatic activities superoxide dismutase (SOD), catalase (CAT), and phenylalanine ammonia-lyase (PAL) with SOD activity increasing up to 7.5-fold under 1 mg/L ethephon + 1 mg/L NAA. Non-enzymatic antioxidants, including total phenolics and FRAP values, were also elevated, indicating improved redox homeostasis. Correlation analysis showed strong positive relationships (r > 0.85) between galanthamine levels, bulblet proliferation, and antioxidant capacity, suggesting coordinated metabolic and morphogenic responses. Interestingly, enzymatic antioxidants exhibited inverse correlations with alkaloid content. These findings demonstrate the synergistic effect of ethephon and IBA in enhancing both biomass and secondary metabolite production. The optimized protocol provides an efficient and sustainable platform for in vitro propagation and high-value alkaloid biosynthesis in L. aestivum, contributing to both conservation and pharmaceutical applications.