Green fluorescent protein (GFP) has been widely used for monitoring gene expression and protein localization in diverse organisms. However, highly sensitive imaging equipment, like fluorescence microscope, is usually required for the visualization of GFP, limitings its application to fixed locations in samples. A reporter that can be visualized in real-time regardless the shape, size and location of the target samples will increase the flexibility and efficiency of research work. Here, we report the application of a GFP-like protein, called eYGFPuv, in both transient expression and stable transformation, in two herbaceous plant species (Arabidopsis and tobacco) and two woody plant species (poplar and citrus). We observed bright fluorescence under UV light in all of the four plant species without any effects on plant growth or development. eYGFPuv was shown to be effective for imaging transient expression in leaf and root tissues. With a focus on in vitro transformation, we demonstrated that the transgenic events expressing 1x eYGFPuv could be easily identified visually during the callus stage and the shoot stage, enabling early and efficient selection of transformants. Furthermore, whole-plant level visualization of eYGFPuv revealed its ubiquitous stability in transgenic plants. In addition, our transformation experiments showed that eYGFPuv can also be used to select transgenic plants without antibiotics. This work demonstrates the feasibility of utilizing 1x eYGFPuv in studies of gene expression and plant transformation in diverse plants.

中文翻译：

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扩大紫外可见报告基因在植物瞬时基因表达和稳定转化中的应用

绿色荧光蛋白 (GFP) 已广泛用于监测不同生物体中的基因表达和蛋白质定位。然而，GFP 的可视化通常需要高灵敏度的成像设备，如荧光显微镜，这限制了其在样品中固定位置的应用。无论目标样本的形状、大小和位置如何，都能实时可视化的报告器将增加研究工作的灵活性和效率。在这里，我们报告了一种名为 eYGFPuv 的类 GFP 蛋白在两种草本植物物种中的瞬时表达和稳定转化的应用。拟南芥和烟草）和两种木本植物（杨树和柑橘）。我们在所有四种植物物种的紫外光下观察到明亮的荧光，对植物生长或发育没有任何影响。eYGFPuv 被证明对叶和根组织中的瞬时表达成像有效。以体外转化为重点，我们证明了表达 1x 的转基因事件eYGFPuv在愈伤组织阶段和发芽阶段可以很容易地在视觉上识别，从而能够早期有效地选择转化体。此外，eYGFPuv 的全植物水平可视化揭示了其在转基因植物中普遍存在的稳定性。此外，我们的转化实验表明，eYGFPuv 也可用于选择不含抗生素的转基因植物。这项工作证明了利用 1x 的可行性eYGFPuv在不同植物中的基因表达和植物转化研究中。