Reverse genetics approaches have revolutionized plant biology and agriculture. Phenomics has the prospect of bridging plant phenotypes with genes, including transgenes, to transform agricultural fields. Genetically encoded fluorescent proteins (FPs) have revolutionized plant biology paradigms in gene expression, protein trafficking and plant physiology. While the first instance of plant canopy imaging of green fluorescent protein (GFP) was performed over 25 years ago, modern phenomics has largely ignored fluorescence as a transgene expression device despite the burgeoning FP colour palette available to plant biologists. Here, we show a new platform for stand‐off imaging of plant canopies expressing a wide variety of FP genes. The platform—the fluorescence‐inducing laser projector (FILP)—uses an ultra‐low‐noise camera to image a scene illuminated by compact diode lasers of various colours, coupled with emission filters to resolve individual FPs, to phenotype transgenic plants expressing FP genes. Each of the 20 FPs screened in plants were imaged at >3 m using FILP in a laboratory‐based laser range. We also show that pairs of co‐expressed fluorescence proteins can be imaged in canopies. The FILP system enabled a rapid synthetic promoter screen: starting from 2000 synthetic promoters transfected into protoplasts to FILP‐imaged agroinfiltrated Nicotiana benthamiana plants in a matter of weeks, which was useful to characterize a water stress‐inducible synthetic promoter. FILP canopy imaging was also accomplished for stably transformed GFP potato and in a split‐GFP assay, which illustrates the flexibility of the instrument for analysing fluorescence signals in plant canopies.

中文翻译：

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冠层中多种荧光蛋白的成像可实现植物的合成生物学

反向遗传学方法彻底改变了植物生物学和农业。物候学具有将植物表型与包括转基因在内的基因桥接以转化农业领域的前景。遗传编码的荧光蛋白（FPs）在基因表达，蛋白运输和植物生理学方面彻底改变了植物生物学范式。尽管25年前对植物冠层进行绿色荧光蛋白（GFP）的首次成像，但现代植物学已将荧光作为转基因表达装置而在很大程度上忽略了荧光，尽管植物生物学家可以使用迅速发展的FP调色板。在这里，我们展示了一个新的平台，用于对表达多种FP基因的植物冠层进行远距离成像。该平台-荧光诱导激光投影仪（FILP）-使用超低噪声摄像头对由各种颜色的紧凑型二极管激光器照明的场景成像，并结合发射滤光片以解析单个FP，以表达表达FP基因的表型转基因植物。使用FILP在基于实验室的激光范围内对植物中筛选的20个FP中的每一个进行了大于3 m的成像。我们还表明，共表达的荧光蛋白对可以在冠层中成像。FILP系统实现了快速的合成启动子筛选：从转染到原生质体中的2000个合成启动子开始到FILP成像的农业浸润 在实验室激光范围内使用FILP 3 m。我们还表明，共表达的荧光蛋白对可以在冠层中成像。FILP系统实现了快速的合成启动子筛选：从转染到原生质体中的2000个合成启动子到FILP成像的农杆菌浸润 在实验室激光范围内使用FILP 3 m。我们还显示，共表达的荧光蛋白对可以在冠层中成像。FILP系统实现了快速的合成启动子筛选：从转染到原生质体中的2000个合成启动子到FILP成像的农杆菌浸润烟草（Nicotiana benthamiana）植物在短短几周内对表征水分胁迫诱导的合成启动子很有用。FILP冠层成像还用于稳定转化的GFP马铃薯以及在split-GFP分析中完成，这说明了用于分析植物冠层荧光信号的仪器的灵活性。