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<p><i>Phlomis bracteosa</i> is an essential medicinal herb native to the Himalayas and is currently facing an alarming decline in its natural populations due to habitat loss and unsustainable harvesting from the wild. To ensure its conservation and sustainable utilization, the current study was conducted to devise an efficient and reproducible <i>in vitro</i> regeneration protocol for <i>P. bracteosa</i> using Murashige and Skoog (MS) medium supplemented with various types, combinations, and concentrations of plant growth regulators (PGRs). In order to analyze shoot organogenesis and somatic embryogenesis from <i>in vitro</i>–derived young leaf and nodal explants, seed germination was optimized by seed coat removal, achieving 100% germination and the shortest mean germination time (MGT) of 7.25 ± 1.53 d on half-strength MS medium with 12.0&#xa0;µM gibberellic acid (GA<sub>3</sub>). Axillary shoot proliferation was maximized with a shoot proliferation coefficient (SPC) of 5.80 ± 0.11 when <i>in vitro</i>–derived nodal explants were cultured on MS medium supplemented with 10.0&#xa0;µM meta-topolin (mT). For adventitious shoot organogenesis 7.5&#xa0;µM benzyladenine (BA) and 2.5&#xa0;µM thidiazuron (TDZ) were most effective for leaf and nodal explants, inducing a maximum mean of 8.42 ± 0.17 and 5.00 ± 0.10 direct shoots per explant, respectively. For somatic embryogenesis, leaf explants treated with a cocktail of 7.5&#xa0;µM BA and 1.0&#xa0;µM indole-3-butyric acid (IBA) induced a peak mean number of 27.5 ± 0.49 embryos and 25.5 ± 0.46 recovered emblings per explant. On the other hand, for nodal explants, 12.5&#xa0;µM 6-(γ,γ-dimethylallylamino) purine (2-iP) proved most potent, yielding 25.50 ± 8.04 embryos and 24.50 ± 0.25 germinating emblings. Microshoots and germinated somatic embryos inoculated on MS medium fortified with 1.0&#xa0;µM IBA exhibited 100% rooting, with an average of 8.20 ± 0.16 roots per microshoot or embling. Well-rooted plantlets acclimatized in Soilrite and under greenhouse conditions showed a 100% survival rate. This study provided a preliminary step to investigate the feasibility of shoot organogenesis and somatic embryogenesis for a scalable and cost-effective mass propagation of <i>P. bracteosa</i>.</p>

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Optimizing efficient and reproducible in vitro regeneration via shoot organogenesis and somatic embryogenesis from leaf and nodal explants of Phlomis bracteosa Royle ex Benth.—a highly valued medicinal herb of the Himalayas

  • 期刊论文

  • 发表日期：

    2026年2月3日

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  • Aashiq Yousuf Bhat,
  • Anwar Shahzad

摘要

Phlomis bracteosa is an essential medicinal herb native to the Himalayas and is currently facing an alarming decline in its natural populations due to habitat loss and unsustainable harvesting from the wild. To ensure its conservation and sustainable utilization, the current study was conducted to devise an efficient and reproducible in vitro regeneration protocol for P. bracteosa using Murashige and Skoog (MS) medium supplemented with various types, combinations, and concentrations of plant growth regulators (PGRs). In order to analyze shoot organogenesis and somatic embryogenesis from in vitro–derived young leaf and nodal explants, seed germination was optimized by seed coat removal, achieving 100% germination and the shortest mean germination time (MGT) of 7.25 ± 1.53 d on half-strength MS medium with 12.0 µM gibberellic acid (GA3). Axillary shoot proliferation was maximized with a shoot proliferation coefficient (SPC) of 5.80 ± 0.11 when in vitro–derived nodal explants were cultured on MS medium supplemented with 10.0 µM meta-topolin (mT). For adventitious shoot organogenesis 7.5 µM benzyladenine (BA) and 2.5 µM thidiazuron (TDZ) were most effective for leaf and nodal explants, inducing a maximum mean of 8.42 ± 0.17 and 5.00 ± 0.10 direct shoots per explant, respectively. For somatic embryogenesis, leaf explants treated with a cocktail of 7.5 µM BA and 1.0 µM indole-3-butyric acid (IBA) induced a peak mean number of 27.5 ± 0.49 embryos and 25.5 ± 0.46 recovered emblings per explant. On the other hand, for nodal explants, 12.5 µM 6-(γ,γ-dimethylallylamino) purine (2-iP) proved most potent, yielding 25.50 ± 8.04 embryos and 24.50 ± 0.25 germinating emblings. Microshoots and germinated somatic embryos inoculated on MS medium fortified with 1.0 µM IBA exhibited 100% rooting, with an average of 8.20 ± 0.16 roots per microshoot or embling. Well-rooted plantlets acclimatized in Soilrite and under greenhouse conditions showed a 100% survival rate. This study provided a preliminary step to investigate the feasibility of shoot organogenesis and somatic embryogenesis for a scalable and cost-effective mass propagation of P. bracteosa.