Group of Phloem Development and Function, Institute of Molecular Plant Biology, Department of Biology, ETH Zürich, Zürich, Switzerland.
Group of Plant Vascular Development, Institute of Molecular Plant Biology, Department of Biology, ETH Zürich, Zürich, Switzerland.
Methods Mol Biol. 2024;2722:3-15. doi: 10.1007/978-1-0716-3477-6_1.
Fluorescent dyes are often used to observe transport mechanisms in plant vascular tissues. However, it has been technically challenging to apply fluorescent dyes on roots to monitor xylem transport in vivo. Here, we present a fast, noninvasive, and high-throughput protocol to monitor xylem transport in seedlings. Using the fluorescent dyes 5(6)-carboxyfluorescein diacetate (CFDA) and Rhodamine WT, we were able to observe xylem transport on a cellular level in Arabidopsis thaliana roots. We describe how to apply these dyes on primary roots of young seedlings, how to monitor root-to-shoot xylem transport, and how to measure xylem transport velocity in roots. Moreover, we show that our protocol can also be applied to lateral roots and grafted seedlings to assess xylem (re)connection. Altogether, these techniques are useful for investigating xylem functionality in diverse experimental setups.
荧光染料常用于观察植物维管组织中的运输机制。然而，将荧光染料应用于根部以监测活体木质部运输在技术上具有挑战性。在这里，我们提出了一种快速、非侵入性和高通量的监测幼苗木质部运输的方案。使用荧光染料 5(6)-羧基荧光素二乙酸酯 (CFDA) 和 Rhodamine WT，我们能够在拟南芥根细胞水平上观察木质部运输。我们描述了如何在幼苗的主根上应用这些染料，如何监测根到梢的木质部运输，以及如何测量根中的木质部运输速度。此外，我们还表明，我们的方案也可以应用于侧根和嫁接幼苗，以评估木质部（再）连接。总之，这些技术对于在不同的实验设置中研究木质部功能非常有用。
