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<p><i>Dendrobium huoshanense</i> Z. Z. Tang &amp; S. J. Cheng is a rare medicinal orchid characterized by slow growth and low propagation efficiency. This study aimed to establish a comprehensive and efficient in vitro propagation system for <i>D. huoshanense</i> by optimizing culture media via a response surface methodology (RSM). Key factors, including plant growth regulators and organic additives, were optimized for distinct developmental stages. The results indicated that the optimal conditions for protocorm proliferation were 0.2&#xa0;mg·L⁻¹ naphthaleneacetic acid (NAA), 1.1&#xa0;mg·L⁻¹ 6-benzylaminopurine (6-BA), and 97.0&#xa0;g·L⁻¹ potato homogenate (PH), resulting in a proliferation coefficient of 59.3. The optimal conditions for protocorm differentiation were 1.1&#xa0;mg·L⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D), 1.1&#xa0;mg·L⁻¹ 6-BA, and 85.0&#xa0;g·L⁻¹ banana homogenate (BH), with an average of 6.4 buds per explant. For protocorm-like body (PLB) proliferation, the optimal conditions were 0.3&#xa0;mg·L⁻¹ NAA, 2.0&#xa0;mg·L⁻¹ kinetin (KT), and 150.0&#xa0;g·L⁻¹ PH, resulting in a proliferation coefficient of 55.2. The optimal medium for PLB differentiation comprised 1.1&#xa0;mg·L⁻¹ 2,4-D, 1.6&#xa0;mg·L⁻¹ KT, and 140.0&#xa0;g·L⁻¹ PH, yielding an average of 7.4 buds per explant. The optimal conditions for the proliferation of cluster shoots were 0.3&#xa0;mg·L⁻¹ 2,4-D, 1.0&#xa0;mg·L⁻¹ KT, 1.6&#xa0;mg·L⁻¹ melatonin (MT), and 150.0&#xa0;g·L⁻¹ BH, resulting in a proliferation coefficient of 6.2. For seedling rooting, the optimal medium was supplemented with 1.4&#xa0;mg·L⁻¹ NAA, 0.7&#xa0;mg·L⁻¹ indole-3-butyric acid (IBA), and 90.0&#xa0;g·L⁻¹ BH. This study not only provides a practical protocol for its rapid propagation and conservation but also a valuable theoretical framework for tissue culture of other endangered orchid species.</p>

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Enhancing Dendrobium huoshanense Z. Z. Tang & S. J. Cheng tissue culture: a response surface methodology approach for robust plantlet regeneration

  • 期刊论文

  • 发表日期：

    2026年2月13日

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  • Yan Zhou,
  • Shixiangtao Ran,
  • Xiaoxiao Wang,
  • Weiting Huang,
  • Zhongming Fang

摘要

Dendrobium huoshanense Z. Z. Tang & S. J. Cheng is a rare medicinal orchid characterized by slow growth and low propagation efficiency. This study aimed to establish a comprehensive and efficient in vitro propagation system for D. huoshanense by optimizing culture media via a response surface methodology (RSM). Key factors, including plant growth regulators and organic additives, were optimized for distinct developmental stages. The results indicated that the optimal conditions for protocorm proliferation were 0.2 mg·L⁻¹ naphthaleneacetic acid (NAA), 1.1 mg·L⁻¹ 6-benzylaminopurine (6-BA), and 97.0 g·L⁻¹ potato homogenate (PH), resulting in a proliferation coefficient of 59.3. The optimal conditions for protocorm differentiation were 1.1 mg·L⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D), 1.1 mg·L⁻¹ 6-BA, and 85.0 g·L⁻¹ banana homogenate (BH), with an average of 6.4 buds per explant. For protocorm-like body (PLB) proliferation, the optimal conditions were 0.3 mg·L⁻¹ NAA, 2.0 mg·L⁻¹ kinetin (KT), and 150.0 g·L⁻¹ PH, resulting in a proliferation coefficient of 55.2. The optimal medium for PLB differentiation comprised 1.1 mg·L⁻¹ 2,4-D, 1.6 mg·L⁻¹ KT, and 140.0 g·L⁻¹ PH, yielding an average of 7.4 buds per explant. The optimal conditions for the proliferation of cluster shoots were 0.3 mg·L⁻¹ 2,4-D, 1.0 mg·L⁻¹ KT, 1.6 mg·L⁻¹ melatonin (MT), and 150.0 g·L⁻¹ BH, resulting in a proliferation coefficient of 6.2. For seedling rooting, the optimal medium was supplemented with 1.4 mg·L⁻¹ NAA, 0.7 mg·L⁻¹ indole-3-butyric acid (IBA), and 90.0 g·L⁻¹ BH. This study not only provides a practical protocol for its rapid propagation and conservation but also a valuable theoretical framework for tissue culture of other endangered orchid species.