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<p><i>Coleus forskohlii</i> Briq. (syn. <i>Plectranthus barbatus</i>), the sole natural source of the diterpenoid forskolin, widely used in pharmaceutical and nutraceutical industries, exhibits low regeneration and chemotypic variability, necessitating robust biotechnological interventions. In this study, an efficient in vitro regeneration and <i>Agrobacterium tumefaciens</i> mediated transformation protocol was standardised. Murashige and Skoog (MS) medium supplemented with 1.0 mg/L BAP and 0.5 mg/L NAA resulted in the highest shoot regeneration frequency (95.54%), while rooting was optimized on half-strength MS medium containing 1.0 mg/L IBA. Genetic transformation was achieved using <i>Agrobacterium tumefaciens</i> harbouring the GUS and RUBY reporter constructs. Response Surface Methodology (RSM) was employed to refine key variables, revealing acetosyringone concentration, bacterial density, and cefotaxime interaction as significant determinants of transformation efficiency. This study reports the first successful stable transformation and plant regeneration using the RUBY reporter in <i>C. forskohlii</i> and establishes a reliable regeneration-transformation platform for future metabolic engineering aimed at enhancing forskolin production.</p>

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Optimization of in vitro regeneration and Agrobacterium mediated transformation in Coleus forskohlii for crop improvement

  • 期刊论文

  • 发表日期：

    2026年2月9日

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  • H. Yashaswini,
  • Shraddha Singh,
  • Sabika Akram,
  • Dinesh A. Nagegegowda,
  • Neelam Prabha Negi

摘要

Coleus forskohlii Briq. (syn. Plectranthus barbatus), the sole natural source of the diterpenoid forskolin, widely used in pharmaceutical and nutraceutical industries, exhibits low regeneration and chemotypic variability, necessitating robust biotechnological interventions. In this study, an efficient in vitro regeneration and Agrobacterium tumefaciens mediated transformation protocol was standardised. Murashige and Skoog (MS) medium supplemented with 1.0 mg/L BAP and 0.5 mg/L NAA resulted in the highest shoot regeneration frequency (95.54%), while rooting was optimized on half-strength MS medium containing 1.0 mg/L IBA. Genetic transformation was achieved using Agrobacterium tumefaciens harbouring the GUS and RUBY reporter constructs. Response Surface Methodology (RSM) was employed to refine key variables, revealing acetosyringone concentration, bacterial density, and cefotaxime interaction as significant determinants of transformation efficiency. This study reports the first successful stable transformation and plant regeneration using the RUBY reporter in C. forskohlii and establishes a reliable regeneration-transformation platform for future metabolic engineering aimed at enhancing forskolin production.