Swinnen Gwen, Lizé Eléonore, Loera Sánchez Miguel, Stolz Stéphanie, Soyk Sebastian
Department of Plant Molecular Biology, University of Lausanne, Lausanne, Switzerland.
Plant Biotechnol J. 2025 Jun 22. doi: 10.1111/pbi.70212.
Genome editing has become a routine tool for functionally characterizing plant and animal genomes. However, stable genome editing in plants remains limited by the time- and labour-intensive process of generating transgenic plants, as well as by the efficient isolation of desired heritable edits. In this study, we evaluated the impact of the morphogenic regulator GRF-GIF on plant regeneration and genome editing outcomes in tomato. We demonstrate that expressing a tomato GRF-GIF chimera reliably accelerates the onset of shoot regeneration from callus tissue culture by approximately one month and nearly doubles the number of recovered transgenic plants. Consequently, the GRF-GIF chimera enables the recovery of a broader range of edited haplotypes and simplifies the isolation of mutants harbouring heritable edits, but without markedly interfering with plant growth and development. Based on these findings, we outline strategies that employ basic or advanced diagnostic pipelines for efficient isolation of single- and higher-order mutants in tomato. Our work represents a technical advantage for tomato transformation and genome editing, with potential applications across other Solanaceae species.
基因组编辑已成为对植物和动物基因组进行功能表征的常规工具。然而，植物中的稳定基因组编辑仍然受到生成转基因植物耗时费力过程的限制，以及有效分离所需可遗传编辑的限制。在本研究中，我们评估了形态发生调节因子GRF-GIF对番茄植物再生和基因组编辑结果的影响。我们证明，表达番茄GRF-GIF嵌合体可可靠地将愈伤组织培养中芽再生的起始时间提前约一个月，并使回收的转基因植物数量几乎增加一倍。因此，GRF-GIF嵌合体能够回收更广泛的编辑单倍型，并简化了具有可遗传编辑的突变体的分离，但不会明显干扰植物的生长和发育。基于这些发现，我们概述了采用基本或先进诊断流程在番茄中高效分离单突变体和高阶突变体的策略。我们的工作代表了番茄转化和基因组编辑的技术优势，在其他茄科物种中具有潜在应用。
