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<p>In this study, an efficient <i>Agrobacterium rhizogenes</i>-mediated transformation system was successfully established for two species in the Sedum genus, including the non-hyperaccumulator <i>S. alfredii</i> and the metal hyperaccumulator <i>S. plumbizincicola.</i> Leaf-petiole cuttings of <i>S. alfredii</i> were used as explants for infection <i>A. rhizogenes</i> strain K599 harboring <i>SaCOPT1-GFP</i> fusion protein gene. Transgenic plants can be obtained within two months and identified by PCR amplification of <i>SaCOPT1-GFP</i> transgene, Western blot of GFP-tagged SaCOPT1 protein, histochemical staining of GUS (<i>β</i>-glucuronidase coding gene) and fluorescence microscopy verifying plasma membrane localization of SaCOPT1-GFP. Functional validation demonstrated that the <i>SaCOPT1-GFP</i>-expressing plants significantly enhanced copper accumulation in roots and leaves (1.7- and 1.3-fold increase vs. wild type, respectively). Despite low bud regeneration frequency in leaf-petiole cuttings of <i>S. plumbizincicola</i>, root regeneration and transformation efficiencies were comparable to <i>S. alfredii</i>. Taken together, this tissue culture-free transformation system provides a rapid, stable platform for gene function studies in Sedum, particularly for decoding the molecular mechanisms underlying metal hypertolerance and hyperaccumulation. It further enables biotechnological applications in phytoremediation engineering and stress-resilient crop development.</p>

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Efficient tissue culture-free genetic transformation and its application for study of metal accumulation in Sedum plants

  • 期刊论文

  • 发表日期：

    2026年1月30日

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  • Xiaonuo Guo,
  • Xin Feng,
  • Hanzhi Fan,
  • Zhaoyuan Guo,
  • Haiyan Zhang

摘要

In this study, an efficient Agrobacterium rhizogenes-mediated transformation system was successfully established for two species in the Sedum genus, including the non-hyperaccumulator S. alfredii and the metal hyperaccumulator S. plumbizincicola. Leaf-petiole cuttings of S. alfredii were used as explants for infection A. rhizogenes strain K599 harboring SaCOPT1-GFP fusion protein gene. Transgenic plants can be obtained within two months and identified by PCR amplification of SaCOPT1-GFP transgene, Western blot of GFP-tagged SaCOPT1 protein, histochemical staining of GUS (β-glucuronidase coding gene) and fluorescence microscopy verifying plasma membrane localization of SaCOPT1-GFP. Functional validation demonstrated that the SaCOPT1-GFP-expressing plants significantly enhanced copper accumulation in roots and leaves (1.7- and 1.3-fold increase vs. wild type, respectively). Despite low bud regeneration frequency in leaf-petiole cuttings of S. plumbizincicola, root regeneration and transformation efficiencies were comparable to S. alfredii. Taken together, this tissue culture-free transformation system provides a rapid, stable platform for gene function studies in Sedum, particularly for decoding the molecular mechanisms underlying metal hypertolerance and hyperaccumulation. It further enables biotechnological applications in phytoremediation engineering and stress-resilient crop development.