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<p>Genetic transformation is essential for functional genomics and molecular breeding in soybean, a globally important crop. However, soybean remains recalcitrant to in vitro regeneration, which limits the efficiency and reproducibility of transformation workflows. Building on our previous research on the <i>GmGASA1-like</i> gene in <i>Glycine max</i>, which demonstrated that its overexpression enhances growth, development, and stress responses, this study focuses on optimising key tissue culture steps within an <i>Agrobacterium</i>-mediated transformation pipeline, with particular emphasis on regeneration and selection responses in the soybean cultivar JN74. By systematically refining key stages, including explant preparation, <i>Agrobacterium</i>-mediated infection, co-cultivation, and regeneration, we improved shoot induction, development, and recovery under selection. The results provide a detailed and reproducible workflow and identify critical bottlenecks affecting transformation efficiency. While molecular confirmation of transformation has been demonstrated in our previous study, the present work focuses on optimisation of the early stages of the transformation process. These findings provide practical guidance for laboratories working with recalcitrant soybean genotypes and support the development of more robust transformation pipelines for functional studies and crop improvement. Although the study was conducted on a single genotype (JN74), the optimisation principles identified offer a practical framework that can be adapted for other soybean cultivars.</p>

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Optimisation of cotyledonary node-based tissue culture workflow for Agrobacterium-mediated transformation in soybean (Glycine max)

  • 期刊论文

  • 发表日期：

    2026年5月7日

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  • Mohamed A. S. Khalifa,
  • Qi Zhang,
  • Abdourazak Alio Moussa,
  • Nooral Amin,
  • Peng Jiao,
  • Lu Liu,
  • Piwu Wang

摘要

Genetic transformation is essential for functional genomics and molecular breeding in soybean, a globally important crop. However, soybean remains recalcitrant to in vitro regeneration, which limits the efficiency and reproducibility of transformation workflows. Building on our previous research on the GmGASA1-like gene in Glycine max, which demonstrated that its overexpression enhances growth, development, and stress responses, this study focuses on optimising key tissue culture steps within an Agrobacterium-mediated transformation pipeline, with particular emphasis on regeneration and selection responses in the soybean cultivar JN74. By systematically refining key stages, including explant preparation, Agrobacterium-mediated infection, co-cultivation, and regeneration, we improved shoot induction, development, and recovery under selection. The results provide a detailed and reproducible workflow and identify critical bottlenecks affecting transformation efficiency. While molecular confirmation of transformation has been demonstrated in our previous study, the present work focuses on optimisation of the early stages of the transformation process. These findings provide practical guidance for laboratories working with recalcitrant soybean genotypes and support the development of more robust transformation pipelines for functional studies and crop improvement. Although the study was conducted on a single genotype (JN74), the optimisation principles identified offer a practical framework that can be adapted for other soybean cultivars.