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<p><i>Platycodon grandiflorum</i> is a globally cultivated ornamental flower as well as a widely utilized traditional medicinal plant and food ingredient. Despite its significance, genetic transformation efforts in <i>P. grandiflorum</i> have been largely confined to the production of transgenic hairy roots. In this study, we established and optimized genetic transformation protocols for <i>P. grandiflorum</i>. We optimized transformation conditions using <i>A. tumefaciens</i> strain EHA105 carrying the pCambia3301 plasmid, achieving 6.77% efficiency with leaf explants. Moreover, we developed a transformation system mediated by <i>Agrobacterium rhizogenes</i>, utilizing the ternary vector p3301WusBBM, which contains the morphogenic regulator genes <i>Wus2</i> and <i>ZmBBM2</i>. This approach significantly enhanced hairy root induction and plant regeneration efficiencies to 43.75% and 21.88%, respectively, thereby improving regeneration efficiency, which has been a challenge in <i>P. grandiflorum</i> transformation. Furthermore, with the improved <i>A. rhizogenes</i>-mediated transformation system combined with CRISPR/Cas9 technology, we achieved precise editing of the endogenous <i>P. grandiflorum</i> gene <i>chr2.2745</i>, attaining a genome editing efficiency of 16.70%. This study establishes an efficient platform for genetic transformation and genome editing in <i>P.&#xa0;grandiflorum</i>, enabling advanced functional genomics research and germplasm enhancement.</p>

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An efficient Agrobacterium-mediated transformation system of Platycodon grandiflorum

  • 期刊论文

  • 发表日期：

    2025年5月12日

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  • Xiaoyu Du,
  • Zhennan Jiang,
  • Qinghui Fu,
  • Yunjun Liu,
  • Yuan Yuan

摘要

Platycodon grandiflorum is a globally cultivated ornamental flower as well as a widely utilized traditional medicinal plant and food ingredient. Despite its significance, genetic transformation efforts in P. grandiflorum have been largely confined to the production of transgenic hairy roots. In this study, we established and optimized genetic transformation protocols for P. grandiflorum. We optimized transformation conditions using A. tumefaciens strain EHA105 carrying the pCambia3301 plasmid, achieving 6.77% efficiency with leaf explants. Moreover, we developed a transformation system mediated by Agrobacterium rhizogenes, utilizing the ternary vector p3301WusBBM, which contains the morphogenic regulator genes Wus2 and ZmBBM2. This approach significantly enhanced hairy root induction and plant regeneration efficiencies to 43.75% and 21.88%, respectively, thereby improving regeneration efficiency, which has been a challenge in P. grandiflorum transformation. Furthermore, with the improved A. rhizogenes-mediated transformation system combined with CRISPR/Cas9 technology, we achieved precise editing of the endogenous P. grandiflorum gene chr2.2745, attaining a genome editing efficiency of 16.70%. This study establishes an efficient platform for genetic transformation and genome editing in P. grandiflorum, enabling advanced functional genomics research and germplasm enhancement.