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<p>The absence of efficient genetic transformation in <i>Cerasus humilis</i> (Bge.) Sok<i>.</i> due to limitations of tissue culture methods was addressed by establishing a culture-free protocol using the cut-dip-budding (CDB) system. By leveraging natural root-suckering regeneration and <i>Agrobacterium rhizogenes</i> K599 (harboring GV1300-<i>ChNAC1</i>-enhanced green fluorescent protein vector), key parameters were optimized: bacterial density (OD<sub>600</sub> = 0.6), vacuum infiltration (20&#xa0;min at 0.08&#xa0;MPa), and 100.0&#xa0;µM acetosyringone. Root explants achieved 20.48% stable transformation efficiency, exceeding stem segments (14.94%). Direct shoot regeneration occurred within 15 d post-infection, yielding intact transgenic plants in 1 to 2 mo. Successful transformation was validated through polymerase chain reaction (PCR), enhanced green fluorescent protein fluorescence analysis, and quantitative real-time PCR, demonstrating robust transgene integration and expression. This study pioneers a rapid transformation system exploiting innate organogenic potential, overcoming regenerative bottlenecks in woody plants. The CDB protocol enables accelerated molecular breeding of Rosaceae crops for arid environments.</p>

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CDB-mediated genetic transformation system for Cerasus humilis without tissue culture

  • 期刊论文

  • 发表日期：

    2026年2月26日

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  • Xin Liu,
  • Shaoyu Lang,
  • Qinghua Shan,
  • Buming Dong,
  • Na Liu,
  • Wenzhi Zhou,
  • Xing Shun Song

摘要

The absence of efficient genetic transformation in Cerasus humilis (Bge.) Sok. due to limitations of tissue culture methods was addressed by establishing a culture-free protocol using the cut-dip-budding (CDB) system. By leveraging natural root-suckering regeneration and Agrobacterium rhizogenes K599 (harboring GV1300-ChNAC1-enhanced green fluorescent protein vector), key parameters were optimized: bacterial density (OD600 = 0.6), vacuum infiltration (20 min at 0.08 MPa), and 100.0 µM acetosyringone. Root explants achieved 20.48% stable transformation efficiency, exceeding stem segments (14.94%). Direct shoot regeneration occurred within 15 d post-infection, yielding intact transgenic plants in 1 to 2 mo. Successful transformation was validated through polymerase chain reaction (PCR), enhanced green fluorescent protein fluorescence analysis, and quantitative real-time PCR, demonstrating robust transgene integration and expression. This study pioneers a rapid transformation system exploiting innate organogenic potential, overcoming regenerative bottlenecks in woody plants. The CDB protocol enables accelerated molecular breeding of Rosaceae crops for arid environments.