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<p>An efficient indirect somatic embryogenesis protocol was developed for the endangered, terrestrial and medicinal orchid <i>Satyrium nepalense</i> using protocorms. Seeds from mature pods were germinated on eleven different nutritional media. Among these media, full-strength Murashige and Skoog (MS) medium favored the highest seed germination rate (98.67%) and supported successive stages of seedling development. Protocorms derived from seeds were cultured on Murashige and Skoog (MS) medium supplemented with auxins (2,4-D, NAA and IBA) either alone or in combination with the cytokinins (TDZ, BA, CPPU and KIN) to evaluate their effectiveness in inducing embryogenic callus and somatic embryogenesis. The most effective response, 71.33% embryogenic callus (EC) and an average of 25.41 somatic embryos (SEs), was observed on full-strength MS medium supplemented with 1.0&#xa0;mg/l 2,4-D and 1.0&#xa0;mg/l TDZ. Subsequent treatment of the emblings with 2.0&#xa0;mg/l IBA promoted tuber formation (0.42&#xa0;cm) and the highest rooting response (85.88%) with an average root length of 6.66&#xa0;cm. The plantlets derived from somatic embryos were successfully hardened using coco peat, brick fragments, wood pieces and charcoal, and acclimatized in a greenhouse with the survival rate of 65%. Histological comparison of in vitro and ex <i>vitro</i> tubers, roots and leaves showed no morphological difference, confirming the protocol’s ability to maintain homogeneity. This protocol can be adapted for further studies in pharmaceutical research and the conservation of medicinally important allied genera.</p>

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In vitro seed germination and indirect somatic embryogenesis of Satyrium nepalense D. Don, a threatened and medicinally important terrestrial orchid

  • 期刊论文

  • 发表日期：

    2026年4月17日

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  • Rengasamy Anbazhakan,
  • Yu-Xin Yue,
  • Brihaspati Poudel,
  • Xin-Meng Zhu,
  • Neng-Qi Li,
  • Jiang-Yun Gao

摘要

An efficient indirect somatic embryogenesis protocol was developed for the endangered, terrestrial and medicinal orchid Satyrium nepalense using protocorms. Seeds from mature pods were germinated on eleven different nutritional media. Among these media, full-strength Murashige and Skoog (MS) medium favored the highest seed germination rate (98.67%) and supported successive stages of seedling development. Protocorms derived from seeds were cultured on Murashige and Skoog (MS) medium supplemented with auxins (2,4-D, NAA and IBA) either alone or in combination with the cytokinins (TDZ, BA, CPPU and KIN) to evaluate their effectiveness in inducing embryogenic callus and somatic embryogenesis. The most effective response, 71.33% embryogenic callus (EC) and an average of 25.41 somatic embryos (SEs), was observed on full-strength MS medium supplemented with 1.0 mg/l 2,4-D and 1.0 mg/l TDZ. Subsequent treatment of the emblings with 2.0 mg/l IBA promoted tuber formation (0.42 cm) and the highest rooting response (85.88%) with an average root length of 6.66 cm. The plantlets derived from somatic embryos were successfully hardened using coco peat, brick fragments, wood pieces and charcoal, and acclimatized in a greenhouse with the survival rate of 65%. Histological comparison of in vitro and ex vitro tubers, roots and leaves showed no morphological difference, confirming the protocol’s ability to maintain homogeneity. This protocol can be adapted for further studies in pharmaceutical research and the conservation of medicinally important allied genera.