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<p><i>Asarum sieboldii</i> Miq. is an important medicinal plant widely used in traditional East Asian medicine for its analgesic, anti-inflammatory, and antimicrobial properties. However, its large-scale utilization is constrained by slow natural propagation and excessive harvesting from wild populations. To address these challenges, this study reported the first efficient and reproducible <i>in vitro</i> propagation protocol for <i>A. sieboldii</i>. Apical shoot explants were cultured to evaluate the effects of basal media, plant growth regulators, and acclimatization conditions on regeneration efficiency. Full-strength Murashige and Skoog (MS) medium was identified as the most suitable basal medium for culture establishment and shoot development. Cytokinin optimization demonstrated that MS medium supplemented with 1.5&#xa0;mg L<sup>−1</sup> 6-benzylaminopurine (BAP) produced optimal shoot proliferation, yielding 5.8 ± 0.83 shoots and 12.25 ± 1.25 leaves per explant after six wk. Root induction was significantly influenced by auxin type and combination, with the highest rooting efficiency achieved on MS medium containing 0.75&#xa0;mg L<sup>−1</sup> naphthaleneacetic acid (NAA) and 1.0&#xa0;mg L<sup>−1</sup> indole-3-butyric acid (IBA). FT-NIR spectral and multivariate chemometric analyses revealed high biochemical similarity between mother and <i>in vitro</i> regenerated plants, with metabolic variation driven primarily by organ type rather than propagation method. Flow cytometric analysis confirmed the maintenance of genome size and ploidy levels, indicating genetic stability of regenerated plants. Morphological observations further demonstrated that <i>in vitro</i> regenerated plants exhibited phenotypic characteristics comparable to mother plants with no visible abnormalities. Acclimatization was highly successful, achieving a 98% survival rate, which confirmed the physiological competence of regenerated plantlets. Overall, this integrated approach provided a robust platform for the conservation, sustainable utilization, and pharmaceutical exploitation of <i>A. sieboldii</i>.</p>

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A novel in vitro propagation protocol for Asarum sieboldii Miq. using apical shoot segments and FT-NIR-chemometric analysis

  • 期刊论文

  • 发表日期：

    2026年7月14日

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  • Kenneth Happy,
  • Roggers Gang,
  • Joyce Mudondo,
  • Ariranur Haniffadli,
  • Yeongjun Ban,
  • Sungyu Yang,
  • Kyeong-Ok Choi,
  • Youngmin Kang

摘要

Asarum sieboldii Miq. is an important medicinal plant widely used in traditional East Asian medicine for its analgesic, anti-inflammatory, and antimicrobial properties. However, its large-scale utilization is constrained by slow natural propagation and excessive harvesting from wild populations. To address these challenges, this study reported the first efficient and reproducible in vitro propagation protocol for A. sieboldii. Apical shoot explants were cultured to evaluate the effects of basal media, plant growth regulators, and acclimatization conditions on regeneration efficiency. Full-strength Murashige and Skoog (MS) medium was identified as the most suitable basal medium for culture establishment and shoot development. Cytokinin optimization demonstrated that MS medium supplemented with 1.5 mg L−1 6-benzylaminopurine (BAP) produced optimal shoot proliferation, yielding 5.8 ± 0.83 shoots and 12.25 ± 1.25 leaves per explant after six wk. Root induction was significantly influenced by auxin type and combination, with the highest rooting efficiency achieved on MS medium containing 0.75 mg L−1 naphthaleneacetic acid (NAA) and 1.0 mg L−1 indole-3-butyric acid (IBA). FT-NIR spectral and multivariate chemometric analyses revealed high biochemical similarity between mother and in vitro regenerated plants, with metabolic variation driven primarily by organ type rather than propagation method. Flow cytometric analysis confirmed the maintenance of genome size and ploidy levels, indicating genetic stability of regenerated plants. Morphological observations further demonstrated that in vitro regenerated plants exhibited phenotypic characteristics comparable to mother plants with no visible abnormalities. Acclimatization was highly successful, achieving a 98% survival rate, which confirmed the physiological competence of regenerated plantlets. Overall, this integrated approach provided a robust platform for the conservation, sustainable utilization, and pharmaceutical exploitation of A. sieboldii.