Although multiple microscopic techniques have been applied to horticultural research, few studies of individual organelles in living fruit cells have been reported to date. In this paper, we established an efficient system for the transient transformation of citrus fruits using an Agrobacterium-mediated method. Kumquat (Fortunella crassifolia Swingle) was used; it exhibits higher transformation efficiency than all citrus fruits that have been tested and a prolonged-expression window. Fruits were transformed with fluorescent reporters, and confocal microscopy and live-cell imaging were used to study their localization and dynamics. Moreover, various pH sensors targeting different subcellular compartments were expressed, and the local pH environments in cells from different plant tissues were compared. The results indicated that vacuoles are most likely the main organelles that contribute to the low pH of citrus fruits. In summary, our method is effective for studying various membrane trafficking events, protein localization, and cell physiology in fruit and can provide new insight into fruit biology research.

中文翻译：

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照亮细胞：柑橘的瞬时转化研究与果实品质相关的基因功能和细胞器活动

尽管多种显微镜技术已应用于园艺研究，但迄今为止很少有关于活水果细胞中单个细胞器的研究报道。在本文中，我们建立了一个高效的柑橘类水果瞬时转化系统，使用农杆菌介导的方法。金橘（金莲花Swingle) 被使用；它表现出比所有已测试的柑橘类水果更高的转化效率和延长的表达窗口。水果被荧光报告基因转化，共聚焦显微镜和活细胞成像被用来研究它们的定位和动力学。此外，还表达了针对不同亚细胞区室的各种 pH 传感器，并比较了来自不同植物组织的细胞中的局部 pH 环境。结果表明，液泡最有可能是导致柑橘类水果低 pH 值的主要细胞器。总之，我们的方法对于研究水果中的各种膜运输事件、蛋白质定位和细胞生理学是有效的，并且可以为水果生物学研究提供新的见解。