SummaryGenetic transformation is a critical tool for gene editing and genetic improvement of plants. Although many model plants and crops can be genetically manipulated, genetic transformation systems for fruit trees are either lacking or perform poorly. We used Rhizobium rhizogenes to transfer the target gene into the hairy roots of Malus domestica and Actinidia chinensis. Transgenic roots were generated within 3 weeks, with a transgenic efficiency of 78.8%. Root to shoot conversion of transgenic hairy roots was achieved within 11 weeks, with a regeneration efficiency of 3.3%. Finally, the regulatory genes involved in stem cell activity were used to improve shoot regeneration efficiency. MdWOX5 exhibited the most significant effects, as it led to an improved regeneration efficiency of 20.6% and a reduced regeneration time of 9 weeks. Phenotypes of the overexpression of RUBY system mediated red roots and overexpression of MdRGF5 mediated longer root hairs were observed within 3 weeks, suggesting that the method can be used to quickly screen genes that influence root phenotype scores through root performance, such as root colour, root hair, and lateral root. Obtaining whole plants of the RUBY system and MdRGF5 overexpression lines highlights the convenience of this technology for studying gene functions in whole plants. Overall, we developed an optimized method to improve the transformation efficiency and stability of transformants in fruit trees.

中文翻译：

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果树中发根根瘤菌介导的基于转基因毛状根向芽转化的高效遗传转化系统

摘要遗传转化是植物基因编辑和遗传改良的重要工具。尽管许多模型植物和作物可以进行基因操纵，但果树的遗传转化系统要么缺乏，要么表现不佳。我们用了发根根瘤菌将目的基因转移到毛状根中海棠和中华猕猴桃。 3周内产生转基因根，转基因效率为78.8%。转基因毛状根在11周内实现从根到芽的转化，再生效率为3.3%。最后，涉及干细胞活性的调控基因被用来提高芽再生效率。MdWOX5表现出最显着的效果，因为它使再生效率提高了 20.6%，再生时间缩短了 9 周。过度表达的表型红宝石系统介导的红根和过度表达中性RGF53周内观察到介导的较长根毛，表明该方法可用于快速筛选通过根性能（例如根颜色、根毛和侧根）影响根表型评分的基因。获得完整植物红宝石系统和中性RGF5过表达系凸显了该技术在研究整个植物基因功能方面的便利性。总的来说，我们开发了一种优化方法来提高果树转化子的转化效率和稳定性。