Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Sakai, Osaka, Japan.
Institute of Transformative Bio-Molecules (WPI-ITbM), Nagoya University, Nagoya, Japan.
Methods Mol Biol. 2020;2177:1-13. doi: 10.1007/978-1-0716-0767-1_1.
Plants take up inorganic nutrients from the soil by transport proteins located in the plasma membrane of root cells. Boron (B) is an essential element for plant growth; it taken up and translocated by boric acid channels such as NIP5;1 and borate exporters such as BOR1 in Arabidopsis. NIP5;1 and BOR1 are localized to the plasma membrane of various root cells in polar manners toward soil- and stele-side, respectively, for efficient transport of B. In response to elevated B concentration, BOR1 undergoes vacuolar sorting for degradation to avoid accumulation of B to a toxic level in tissues. The polar localization and vacuolar sorting of the transport proteins are regulated through differential mechanisms of endocytosis and intracellular trafficking. In this chapter, we describe methods for quantitative live-cell imaging of GFP-NIP5;1 and BOR1-GFP as markers for the polar and vacuolar trafficking.
植物通过位于根细胞质膜上的转运蛋白从土壤中摄取无机养分。硼（B）是植物生长所必需的元素；它被硼酸通道如 NIP5;1 和硼酸盐外排泵如拟南芥中的 BOR1 摄取和转运。NIP5;1 和 BOR1 分别以极性方式定位于各种根细胞的质膜上，朝向土壤和中柱侧，以实现 B 的有效运输。为了避免 B 在组织中积累到毒性水平，BOR1 在受到高 B 浓度的刺激后会进行液泡分拣，从而降解。这些转运蛋白的极性定位和液泡分拣是通过内吞作用和细胞内运输的不同机制来调节的。在本章中，我们描述了使用 GFP-NIP5;1 和 BOR1-GFP 作为极性和液泡运输的标记进行定量活细胞成像的方法。
