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<p><i>Cannabis sativa</i> L. comprises &gt; 500 compounds, including cannabidiol (CBD), which have multiple pharmacological benefits such as anti-inflammatory, antioxidant, and neuroprotective influence. It is suggested that CBD accumulates in the adventitious roots (ARs) of <i>C. sativa.</i> This study systematically optimized the induction and liquid culture parameters for <i>C. sativa</i> ARs and assessed the capability of the endophytic fungus RF-1 to enhance the synthesis of flavonoids, phenolics, and CBD. According to the results, the optimal medium for ARs induction was sucrose (30&#xa0;g/L) + 1-Naphthylacetic acid (NAA; 2.0&#xa0;mg/L) + Indole-3-butyric acid (IBA; 1.0&#xa0;mg/L) + Murashige and Skoog (MS). Furthermore, the optimized liquid culture system included MS + IBA (0.5&#xa0;mg/L) + NAA (1.0&#xa0;mg/L), with a 30-day growth cycle. Moreover, ARs induced with RF-1 fermentation broth indicated the highest permeability, as well as superoxide dismutase and peroxidase enzyme activities, while the lowest Malondialdehyde content, indicating minimal cellular oxidative damage. Moreover, prenyltransferase and phenylalanine ammonia-lyase enzyme activities, and their gene expression levels were significantly elevated, resulting in the highest yield of total phenol, flavonoid, and CBD contents. In conclusion, this study elucidated the optimal conditions for inducing and culturing ARs and investigated the impact of a biotic elicitor on their culture, thus providing novel insights into the synthesis of metabolites and CBD accumulation in vitro.</p>

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Optimization of in vitro culture conditions for Cannabis sativa L. adventitious roots and enhancement of flavonoid, phenolic, and cannabidiol accumulation by endophytic fungus RF-1

  • 期刊论文

  • 发表日期：

    2026年2月17日

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  • JiaXin Shao,
  • Yongxin Wang,
  • Dan Wang,
  • Shuai Li,
  • Ying Zhai,
  • Yan Zhao,
  • TianYi Ma,
  • ZhiGang Wang,
  • Ming Zhao,
  • ShanShan Li

摘要

Cannabis sativa L. comprises > 500 compounds, including cannabidiol (CBD), which have multiple pharmacological benefits such as anti-inflammatory, antioxidant, and neuroprotective influence. It is suggested that CBD accumulates in the adventitious roots (ARs) of C. sativa. This study systematically optimized the induction and liquid culture parameters for C. sativa ARs and assessed the capability of the endophytic fungus RF-1 to enhance the synthesis of flavonoids, phenolics, and CBD. According to the results, the optimal medium for ARs induction was sucrose (30 g/L) + 1-Naphthylacetic acid (NAA; 2.0 mg/L) + Indole-3-butyric acid (IBA; 1.0 mg/L) + Murashige and Skoog (MS). Furthermore, the optimized liquid culture system included MS + IBA (0.5 mg/L) + NAA (1.0 mg/L), with a 30-day growth cycle. Moreover, ARs induced with RF-1 fermentation broth indicated the highest permeability, as well as superoxide dismutase and peroxidase enzyme activities, while the lowest Malondialdehyde content, indicating minimal cellular oxidative damage. Moreover, prenyltransferase and phenylalanine ammonia-lyase enzyme activities, and their gene expression levels were significantly elevated, resulting in the highest yield of total phenol, flavonoid, and CBD contents. In conclusion, this study elucidated the optimal conditions for inducing and culturing ARs and investigated the impact of a biotic elicitor on their culture, thus providing novel insights into the synthesis of metabolites and CBD accumulation in vitro.