Despite the transformative power of gene editing for crop improvement, its widespread application across species and varieties is limited by the transformation bottleneck that exists for many crops. The genetic transformation of plants is hindered by a general reliance on in vitro regeneration through plant tissue culture. Tissue culture requires empirically-determined conditions and aseptic techniques, and cannot easily be translated to recalcitrant species and genotypes. Both Agrobacterium-mediated and alternative transformation protocols are limited by a dependency on in vitro regeneration, which also limits their use by non-experts and hinders research into non-model species such as those of possible novel biopharmaceutical or nutraceutical use, as well as novel ornamental varieties. Hence, there is significant interest in developing tissue culture-independent plant transformation and gene editing approaches which can circumvent the bottlenecks associated with in vitro plant regeneration recalcitrance. Compared to tissue culture-based transformations, tissue culture-independent approaches offer advantages such as avoidance of somaclonal variation effects, with more streamlined and expeditious methodological processes. The ease of use, dependability, and accessibility of tissue culture-independent procedures can make them attractive to non-experts, outperforming classic tissue culture-dependent systems. This review explores the diversity of tissue culture-independent transformation approaches and compares them to traditional tissue culture-dependent transformation strategies. We highlight their simplicity and provide examples of recent successful transformations accomplished using these systems. Our review also addresses current limitations and explores future perspectives, highlighting the significance of these techniques for advancing plant research and crop improvement.

中文翻译：

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不依赖组织培养的方法革新植物转化和基因编辑


尽管基因编辑对作物改良具有变革性的力量，但其跨物种和品种的广泛应用受到许多作物存在的转化瓶颈的限制。植物的遗传转化受到普遍依赖通过植物组织培养进行的体外再生的阻碍。组织培养需要根据经验确定的条件和无菌技术，并且不容易转化为顽固的物种和基因型。农杆菌介导的和替代转化方案都受到对体外再生的依赖性的限制，这也限制了非专家对它们的使用，并阻碍了对非模式物种的研究，例如可能用于新型生物制药或营养保健品的物种，以及新的观赏品种。因此，开发不依赖组织培养的植物转化和基因编辑方法具有浓厚的兴趣，这些方法可以规避与体外植物再生顽固相关的瓶颈。与基于组织培养的转化相比，不依赖组织培养的方法具有避免体细胞克隆变异效应等优势，具有更简化和快速的方法过程。独立于组织培养的程序的易用性、可靠性和可及性使其对非专家具有吸引力，优于经典的组织培养依赖系统。本综述探讨了不依赖组织培养的转化方法的多样性，并将其与传统的组织培养依赖性转化策略进行了比较。我们重点介绍了它们的简单性，并提供了最近使用这些系统完成的成功转型的示例。 我们的综述还解决了当前的局限性并探讨了未来的前景，强调了这些技术对推进植物研究和作物改良的重要性。