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<p>The low efficiency of genetic transformation systems has hindered gene function analysis and precise molecular breeding in the tea plant (<i>Camellia sinensis</i>). In this study, we established an efficient <i>Agrobacterium tumefaciens</i>-mediated transformation system for calli of ‘Fuding Dabaicha’. Using stem segments as explants, soft calli with a low polyphenol content were induced on Murashige and Skoog medium supplemented with 2 mg·L<sup>− 1</sup> thidiazuron and 0.2 mg·L<sup>− 1</sup> indole-3-acetic acid in the dark. The callus proliferation rate was 5.99. Transformation efficiency of 77.53% was achieved under the combination of 100 mg·L<sup>− 1</sup> kanamycin, 50 mg·L<sup>− 1</sup> carbenicillin, and 100 µmol·L<sup>− 1</sup> acetosyringone. The <i>CsCLH1</i>-overexpressing transgenic calli were identified by β-glucuronidase histochemical staining, PCR, semi-quantitative reverse transcription–PCR, western blotting, and real-time quantitative PCR. This callus transformation system provides a reliable technical tool for gene function analysis and genetic improvement of tea plants.</p>

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Efficient Agrobacterium tumefaciens-mediated transient transformation of low-polyphenol callus from tea plant (Camellia sinensis (L.) Kuntze)

  • 期刊论文

  • 发表日期：

    2026年6月26日

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  • Yue Wang,
  • Mingli Li,
  • Jiyan Li,
  • Lirong Sun,
  • Hanyue Wang,
  • Tongyan Xin,
  • Jingjing Feng,
  • Yanan Ren,
  • Weimin Liu,
  • Yusheng Li,
  • Min Li

摘要

The low efficiency of genetic transformation systems has hindered gene function analysis and precise molecular breeding in the tea plant (Camellia sinensis). In this study, we established an efficient Agrobacterium tumefaciens-mediated transformation system for calli of ‘Fuding Dabaicha’. Using stem segments as explants, soft calli with a low polyphenol content were induced on Murashige and Skoog medium supplemented with 2 mg·L− 1 thidiazuron and 0.2 mg·L− 1 indole-3-acetic acid in the dark. The callus proliferation rate was 5.99. Transformation efficiency of 77.53% was achieved under the combination of 100 mg·L− 1 kanamycin, 50 mg·L− 1 carbenicillin, and 100 µmol·L− 1 acetosyringone. The CsCLH1-overexpressing transgenic calli were identified by β-glucuronidase histochemical staining, PCR, semi-quantitative reverse transcription–PCR, western blotting, and real-time quantitative PCR. This callus transformation system provides a reliable technical tool for gene function analysis and genetic improvement of tea plants.