The Agrobacterium-mediated transient expression system has been developed and applied to various plants as an alternative to stable transformation. However, its application in tomatoes is still limited due to low expression efficiency. In this study, we describe an improved vacuum-infiltration system that can be used in both tomato fruits and leaves. Notably, this study is the first report of vacuum infiltration in attached tomato fruits. The feasibility of the improved vacuum-infiltration system in Micro-Tom tomato was confirmed by various assays, including multiple fluorescent protein expression analysis, β-glucuronidase activity analysis, and RUBY reporter visualization. Subsequently, the improved vacuum-infiltration system was successfully applied to tomato biotechnology research. Herein, a trichome-specific promoter in tomato was identified that can drive the directional synthesis of specific plant natural products (PNPs). Additionally, based on the assessment results of the improved vacuum-infiltration system, we obtained a flavonoid-rich tomato variety through the stable transformation of AmRosea and AmDelila. In a significant practical application, we successfully synthesized the high-value scutellarin in tomato, which provides an alternative route for the production of PNPs from plants. In addition, the improved vacuum-infiltration system has been demonstrated to be suitable for commercial tomato varieties (‘Emerald’ and ‘Provence’) as well. The improved vacuum-infiltration system not only speeds up fundamental and applied research in tomato but also offers an additional powerful tool for advancing tomato synthetic biology research.

中文翻译：

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真空渗透系统在番茄中的改进与应用


农杆菌介导的瞬时表达系统已被开发并应用于各种植物，作为稳定转化的替代方案。然而，由于表达效率低，它在西红柿中的应用仍然受到限制。在这项研究中，我们描述了一种改进的真空渗透系统，该系统可用于番茄果实和叶子。值得注意的是，这项研究是附着番茄果实真空浸润的首次报道。通过各种测定证实了改进的真空浸润系统在 Micro-Tom 番茄中的可行性，包括多种荧光蛋白表达分析、β-葡萄糖醛酸酶活性分析和 RUBY 报告基因可视化。随后，改进的真空渗透系统成功地应用于番茄生物技术研究。在此，在番茄中鉴定出一种毛状体特异性启动子，它可以驱动特定植物天然产物 （PNP） 的定向合成。此外，基于改进的真空渗透系统的评估结果，我们通过 AmRosea 和 AmDelila 的稳定转化获得了富含类黄酮的番茄品种。在一个重要的实际应用中，我们成功地在番茄中合成了高价值的黄芩素，这为从植物生产 PNP 提供了另一种途径。此外，改进的真空渗透系统已被证明也适用于商业番茄品种（“翡翠”和“普罗旺斯”）。改进的真空渗透系统不仅加快了番茄的基础和应用研究，还为推进番茄合成生物学研究提供了另一个强大的工具。