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<p><i>Chrysanthemum morifolium</i>, an economically important ornamental and medicinal species, faces limitations in genetic improvement due to sporophytic self-incompatibility (SSI), narrow genetic diversity, and susceptibility to biotic and abiotic stresses. This study establishes a highly efficient, callus-free regeneration and transformation system to address these challenges. A direct organogenesis protocol using leaf explants and optimized hormones (2.0&#xa0;mg/L BAP + 0.1&#xa0;mg/L NAA) achieved (92.5% ± 6.61) regeneration efficiency without callus intervention, minimizing somaclonal variation and ensuring genetic fidelity. Furthermore, systematic optimization of <i>Agrobacterium tumefaciens</i>-mediated transformation parameters, bacterial density (OD₆₀₀ 0.6), acetosyringone (200 µM), kanamycin (20&#xa0;mg/L), and infection time (15&#xa0;min), yielded a (84.8% ± 3.96) transformation success rate, with robust survival of acclimatized transgenic plants. Overall, this efficient and reproducible system provides a robust platform for functional genomics, CRISPR/Cas-based genome editing, and targeted trait enhancement, enabling rapid development of stress-resilient, high-value chrysanthemum cultivars for both research and commercial breeding applications.</p>

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Establishment of an efficient plant regeneration and Agrobacterium-mediated transformation protocol in ornamental Chrysanthemum (C. morifolium)

  • 期刊论文

  • 发表日期：

    2026年2月18日

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  • Gazala Parween,
  • Tanya Singh,
  • Zakir Husain,
  • Sabika Akram,
  • Kahkashan Khatoon,
  • Zafar Iqbal Warsi,
  • Parmeshwar Singh,
  • Laiq Ur Rahman

摘要

Chrysanthemum morifolium, an economically important ornamental and medicinal species, faces limitations in genetic improvement due to sporophytic self-incompatibility (SSI), narrow genetic diversity, and susceptibility to biotic and abiotic stresses. This study establishes a highly efficient, callus-free regeneration and transformation system to address these challenges. A direct organogenesis protocol using leaf explants and optimized hormones (2.0 mg/L BAP + 0.1 mg/L NAA) achieved (92.5% ± 6.61) regeneration efficiency without callus intervention, minimizing somaclonal variation and ensuring genetic fidelity. Furthermore, systematic optimization of Agrobacterium tumefaciens-mediated transformation parameters, bacterial density (OD₆₀₀ 0.6), acetosyringone (200 µM), kanamycin (20 mg/L), and infection time (15 min), yielded a (84.8% ± 3.96) transformation success rate, with robust survival of acclimatized transgenic plants. Overall, this efficient and reproducible system provides a robust platform for functional genomics, CRISPR/Cas-based genome editing, and targeted trait enhancement, enabling rapid development of stress-resilient, high-value chrysanthemum cultivars for both research and commercial breeding applications.