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<p><i>Schizonepeta tenuifolia</i> (Benth.) Briq., a medicinal plant rich in the terpene pulegone, is currently an effective clinical treatment for respiratory diseases. To investigate the role of monoterpene synthase in its secondary metabolism, we established, for the first time, a regeneration and <i>Agrobacterium</i>-mediated genetic transformation method for <i>S. tenuifolia</i>. Using sterile seedling nodal segments as explants, the optimal callus induction and adventitious shoot regeneration were achieved on Murashige and Skoog medium supplemented with 0.5&#xa0;mg/L (2.686&#xa0;μM) 1-naphthaleneacetic acid and 1.5&#xa0;mg/L (6.812&#xa0;μM) thidiazuron. Rooting was most effective on 1/2 MS medium with 0.3–0.4&#xa0;mg/L (1.611–2.148&#xa0;μM) NAA. For genetic transformation, optimal conditions included a 10-day pre-culture, an <i>Agrobacterium</i> solution with OD<sub>600</sub> of 0.7, and a 15-min infection period, followed by a 3-day co-culture, significantly enhancing positive transgenic plant rate. GFP fluorescence and PCR analysis confirmed successful exogenous gene integration and expression. Overexpression of <i>StIPI</i>, encoding a monoterpene synthase, increased glandular trichome density on leaves without affecting plant growth and significantly elevated pulegone concentration. This system facilitates functional gene insertion via<i> Agrobacterium</i>-mediated transformation in <i>S. tenuifolia</i>.</p>

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An efficient Schizonepeta tenuifolia transformation system for enhanced pulegone production via StIPI overexpression

  • 期刊论文

  • 发表日期：

    2025年8月5日

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  • Jimeng Zhang,
  • Jianling Shi,
  • Yang Gao,
  • Liqiong Sun,
  • Kangcai Wang,
  • Xiaoqing Tang

摘要

Schizonepeta tenuifolia (Benth.) Briq., a medicinal plant rich in the terpene pulegone, is currently an effective clinical treatment for respiratory diseases. To investigate the role of monoterpene synthase in its secondary metabolism, we established, for the first time, a regeneration and Agrobacterium-mediated genetic transformation method for S. tenuifolia. Using sterile seedling nodal segments as explants, the optimal callus induction and adventitious shoot regeneration were achieved on Murashige and Skoog medium supplemented with 0.5 mg/L (2.686 μM) 1-naphthaleneacetic acid and 1.5 mg/L (6.812 μM) thidiazuron. Rooting was most effective on 1/2 MS medium with 0.3–0.4 mg/L (1.611–2.148 μM) NAA. For genetic transformation, optimal conditions included a 10-day pre-culture, an Agrobacterium solution with OD600 of 0.7, and a 15-min infection period, followed by a 3-day co-culture, significantly enhancing positive transgenic plant rate. GFP fluorescence and PCR analysis confirmed successful exogenous gene integration and expression. Overexpression of StIPI, encoding a monoterpene synthase, increased glandular trichome density on leaves without affecting plant growth and significantly elevated pulegone concentration. This system facilitates functional gene insertion via Agrobacterium-mediated transformation in S. tenuifolia.