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<p><i>Impatiens walleriana</i> has garnered significant attention due to its medicinal, ornamental, and economic value. However, traditional propagation methods (seeds and cuttings) face challenges such as low seed maturity, poor rooting rate, seasonal constraints, and susceptibility to diseases, making it difficult to meet market demands. Current tissue culture techniques rely on axillary bud proliferation, but low proliferation efficiency and difficulties in callus differentiation hinder large scale production and molecular breeding. To overcome these bottlenecks, this study optimized hormone combinations (e.g., 6-BA, TDZ, and NAA), screened efficient explants (hypocotyls with cotyledons), and established a callus induction and adventitious bud regeneration system. On MS medium supplemented with 2.0&#xa0;mg/L 6-BA, 2.5&#xa0;mg/L TDZ, and 0.5&#xa0;mg/L NAA, a callus induction rate of 93.33% was achieved. When using 1.0&#xa0;mg/L 6-BA, 0.2&#xa0;mg/L NAA, and 0.3&#xa0;mg/L IAA, adventitious bud induction peaked at 48.67 buds per explant, while the rooting efficiency reached 76% with 0.6&#xa0;mg/L TDZ and 3&#xa0;mg/L NAA. Combined with <i>Agrobacterium</i>-mediated CRISPR/Cas9 technology targeting MYB gene editing (transformation efficiency of 2.1%), transgenic plants with phenotypic variations were successfully obtained. This system addresses the limitations of traditional propagation and provides technical support for rapid propagation, germplasm preservation, and disease-resistant/stress-tolerant molecular breeding in <i>Impatiens</i> species.</p>

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Establishment of regeneration and CRISPR/Cas9-mediated genetic transformation systems in Impatiens walleriana

  • 期刊论文

  • 发表日期：

    2025年10月16日

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  • Huang Deng,
  • Yulu Wu,
  • Shanman Li,
  • Baiyang He,
  • Yuteng Xue,
  • Shunming Zhang,
  • Xue Hu,
  • Yu Zhou,
  • Jiwei Wang,
  • Yuhang Wang,
  • Songtao He,
  • Yuhong Rong,
  • Chao Luo,
  • Xinxiang Bai

摘要

Impatiens walleriana has garnered significant attention due to its medicinal, ornamental, and economic value. However, traditional propagation methods (seeds and cuttings) face challenges such as low seed maturity, poor rooting rate, seasonal constraints, and susceptibility to diseases, making it difficult to meet market demands. Current tissue culture techniques rely on axillary bud proliferation, but low proliferation efficiency and difficulties in callus differentiation hinder large scale production and molecular breeding. To overcome these bottlenecks, this study optimized hormone combinations (e.g., 6-BA, TDZ, and NAA), screened efficient explants (hypocotyls with cotyledons), and established a callus induction and adventitious bud regeneration system. On MS medium supplemented with 2.0 mg/L 6-BA, 2.5 mg/L TDZ, and 0.5 mg/L NAA, a callus induction rate of 93.33% was achieved. When using 1.0 mg/L 6-BA, 0.2 mg/L NAA, and 0.3 mg/L IAA, adventitious bud induction peaked at 48.67 buds per explant, while the rooting efficiency reached 76% with 0.6 mg/L TDZ and 3 mg/L NAA. Combined with Agrobacterium-mediated CRISPR/Cas9 technology targeting MYB gene editing (transformation efficiency of 2.1%), transgenic plants with phenotypic variations were successfully obtained. This system addresses the limitations of traditional propagation and provides technical support for rapid propagation, germplasm preservation, and disease-resistant/stress-tolerant molecular breeding in Impatiens species.