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<p><i>Zephyranthes splendens</i>, an indigenous Chilean species endemic to the south-central foothills of the Andes, belongs to the subfamily Amaryllidoideae (Amaryllidaceae), well-known for its production of isoquinoline-type bioactive alkaloids. For the first time, we describe an in vitro protocol for the micropropagation and bulbification of <i>Z. splendens</i>, along with detailed characterization of in vitro alkaloid pattern in the resulting tissue cultured. To optimize the in vitro growth of the plant on MS medium, combinations of different sucrose concentrations and growth regulators (BAP and NAA) were evaluated. It is determined that the combination of 2&#xa0;mg/L BAP and 0.2&#xa0;mg/L NAA promoted highly efficient direct organogenesis, while 90&#xa0;g/L sucrose favoured bulb proliferation. Also, kinetin concentration and culture duration were evaluated for their effects on bulb development. Higher concentrations of kinetin significantly increased bulb volume over time, while sucrose enhanced shoot development when influenced by kinetin. Statistical analysis revealed that both kinetin concentration and in vitro culture period were key factors for influencing bulb growth. GC-MS based alkaloid profiling demonstrated quantitative differences between wild bulbs and in vitro derived bulblets, with the latter exhibiting minor metabolic activity and reduced alkaloid diversity. The juvenile vegetative period along with a diminished photosynthetic process, may plays main role in metabolic processes in in vitro bulblets. Overall, these results highlight the ability of <i>Z. splendens</i> bulblets to produce alkaloids under controlled in vitro conditions, providing a methodological basis for their subsequent biotechnological control.</p> Graphical abstract <p></p>

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Effective in vitro bulbification and alkaloid dereplication by GC-MS of Chilean Zephyranthes splendens, an endemic species of the Andes mountains

  • 期刊论文

  • 发表日期：

    2025年8月11日

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  • Ricardo E. Hernández,
  • Jean Paulo de Andrade,
  • Natalia Opazo,
  • Ignacia Castillo,
  • Strahil Berkov,
  • Ariel D. Arencibia

摘要

Zephyranthes splendens, an indigenous Chilean species endemic to the south-central foothills of the Andes, belongs to the subfamily Amaryllidoideae (Amaryllidaceae), well-known for its production of isoquinoline-type bioactive alkaloids. For the first time, we describe an in vitro protocol for the micropropagation and bulbification of Z. splendens, along with detailed characterization of in vitro alkaloid pattern in the resulting tissue cultured. To optimize the in vitro growth of the plant on MS medium, combinations of different sucrose concentrations and growth regulators (BAP and NAA) were evaluated. It is determined that the combination of 2 mg/L BAP and 0.2 mg/L NAA promoted highly efficient direct organogenesis, while 90 g/L sucrose favoured bulb proliferation. Also, kinetin concentration and culture duration were evaluated for their effects on bulb development. Higher concentrations of kinetin significantly increased bulb volume over time, while sucrose enhanced shoot development when influenced by kinetin. Statistical analysis revealed that both kinetin concentration and in vitro culture period were key factors for influencing bulb growth. GC-MS based alkaloid profiling demonstrated quantitative differences between wild bulbs and in vitro derived bulblets, with the latter exhibiting minor metabolic activity and reduced alkaloid diversity. The juvenile vegetative period along with a diminished photosynthetic process, may plays main role in metabolic processes in in vitro bulblets. Overall, these results highlight the ability of Z. splendens bulblets to produce alkaloids under controlled in vitro conditions, providing a methodological basis for their subsequent biotechnological control.

Graphical abstract