Division of Biological and Environmental Science and Engineering, Center for Desert Agriculture, the BioActives Lab, King Abdullah University of Science and Technology, Thuwal, Saudi Arabia.
Institute of Synthetic Biology and CEPLAS, University of Düsseldorf, Düsseldorf, Germany.
Methods Mol Biol. 2021;2309:201-218. doi: 10.1007/978-1-0716-1429-7_16.
Understanding the biological background of strigolactone (SL) structural diversity and the SL signaling pathway at molecular level requires quantitative and sensitive tools that precisely determine SL dynamics. Such biosensors may be also very helpful in screening for SL analogs and mimics with defined biological functions.Recently, the genetically encoded, ratiometric sensor StrigoQuant was developed and allowed the quantification of the activity of a wide concentration range of SLs. StrigoQuant can be used for studies on the biosynthesis, function and signal transduction of this hormone class.Here, we provide a comprehensive protocol for establishing the use of StrigoQuant in Arabidopsis protoplasts. We first describe the generation and transformation of the protoplasts with StrigoQuant and detail the application of the synthetic SL analogue GR24. We then show the recording of the luminescence signal and how the obtained data are processed and used to assess/determine SL perception.
理解独脚金内酯（SL）结构多样性的生物学背景和分子水平上的 SL 信号通路需要定量和敏感的工具，这些工具能精确地确定 SL 的动态。这样的生物传感器在筛选具有特定生物学功能的 SL 类似物和模拟物方面也可能非常有帮助。最近，开发了遗传编码的比率传感器 StrigoQuant，允许定量分析广泛浓度范围内 SL 的活性。StrigoQuant 可用于研究该激素类别的生物合成、功能和信号转导。在这里，我们提供了一个在拟南芥原生质体中建立 StrigoQuant 使用的综合方案。我们首先描述了带有 StrigoQuant 的原生质体的产生和转化，并详细介绍了合成 SL 类似物 GR24 的应用。然后，我们展示了发光信号的记录，以及如何处理获得的数据，并用于评估/确定 SL 的感知。
