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<p>Most in vitro regeneration protocols for <i>Coleus forskohlii</i> are based on nodes and shoot tips. Here, we evaluated transverse thin cell layers (tTCL; ~0.2–0.3&#xa0;mm) derived from leaf, node, and internode to establish a direct organogenesis route and document morpho-anatomical progression. Explants were cultured on Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurine (BAP; 0.4-2.0 mg L<sup>− 1</sup>) or thidiazuron (TDZ; 0.4-2.0 mg L<sup>− 1</sup>). Thidiazuron at 1.2 mg L<sup>− 1</sup> produced the highest regeneration frequency, with node tTCL showing the strongest response (up to 95.8% regeneration; 9.0 ± 1.0 shoots per explant), followed by leaf (82.0%; 6.0 ± 0.6) and internode (77.8%; 4.0 ± 0.0). Weekly microscopic observations indicated epidermal/sub-epidermal origin of shoot primordia in leaf tTCL, whereas node and internode responses were associated with intense divisions in the vascular cambium. Regenerated shoots proliferated, rooted, and acclimatized successfully. This tTCL-based approach provides an efficient platform for rapid clonal propagation and developmental analysis in <i>C. forskohlii</i>. These results provide a foundational framework for future investigations aimed at evaluating important translational parameters, confirming clonal fidelity, and assessing phytochemical consistency.</p>

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Concentration-dependent optimization of cytokinins on direct organogenesis from transverse thin cell layer (tTCL) explants of Coleus forskohlii (Willd.) Briq. – a morpho-anatomical approach for sustainable plant production

  • 期刊论文

  • 发表日期：

    2026年4月7日

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  • M. Manokari,
  • Anantpal Shekhawat,
  • Mohammad Faisal,
  • Abdulrahman A. Alatar,
  • Rupesh Kumar Singh,
  • Mahipal S. Shekhawat

摘要

Most in vitro regeneration protocols for Coleus forskohlii are based on nodes and shoot tips. Here, we evaluated transverse thin cell layers (tTCL; ~0.2–0.3 mm) derived from leaf, node, and internode to establish a direct organogenesis route and document morpho-anatomical progression. Explants were cultured on Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurine (BAP; 0.4-2.0 mg L− 1) or thidiazuron (TDZ; 0.4-2.0 mg L− 1). Thidiazuron at 1.2 mg L− 1 produced the highest regeneration frequency, with node tTCL showing the strongest response (up to 95.8% regeneration; 9.0 ± 1.0 shoots per explant), followed by leaf (82.0%; 6.0 ± 0.6) and internode (77.8%; 4.0 ± 0.0). Weekly microscopic observations indicated epidermal/sub-epidermal origin of shoot primordia in leaf tTCL, whereas node and internode responses were associated with intense divisions in the vascular cambium. Regenerated shoots proliferated, rooted, and acclimatized successfully. This tTCL-based approach provides an efficient platform for rapid clonal propagation and developmental analysis in C. forskohlii. These results provide a foundational framework for future investigations aimed at evaluating important translational parameters, confirming clonal fidelity, and assessing phytochemical consistency.