Basic Forestry and Proteomic Research Center, College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, China.
Basic Forestry and Proteomic Research Center, College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, China.
Methods Cell Biol. 2020;160:253-261. doi: 10.1016/bs.mcb.2020.03.005. Epub 2020 Apr 6.
The microtubule cytoskeleton plays an important role in cell shape and plant development. During the past decades, the ability to use confocal microcopy to observe microtubules in living cells using fluorescent protein fusions has given plant scientists the opportunity to answer outstanding biological questions. Plants contain diverse epidermal cells with distinct morphologies and physiological functions. For example, flowering plants have specialized petal conical cells that likely facilitate functions such as providing grips for bee pollinators. Here, we summarize recent progress on live imaging of the microtubule cytoskeleton in conical cells. Firstly, we present a simple method for live-cell confocal imaging of conical cells, which is suitable for the quantification of the cell geometry. Secondly, we describe a method for observing microtubule organization in conical cells of Arabidopsis thaliana expressing green fluorescent protein (GFP)-tagged α-tubulin 6 (GFP-TUA6). These live imaging approaches are likely to lead to rapid advances in our knowledge of the role of microtubules in conical cell shaping.
微管细胞骨架在细胞形态和植物发育中发挥着重要作用。在过去的几十年中，使用荧光蛋白融合物通过共焦显微镜观察活细胞中的微管的能力使植物科学家有机会回答悬而未决的生物学问题。植物含有具有不同形态和生理功能的各种表皮细胞。例如，开花植物具有专门的花瓣锥形细胞，这些细胞可能有助于为蜜蜂传粉者提供抓握力等功能。在这里，我们总结了在锥形细胞中对微管细胞骨架进行活体成像的最新进展。首先，我们提出了一种用于锥形细胞活细胞共焦成像的简单方法，该方法适用于细胞几何形状的定量。其次，我们描述了一种用于观察拟南芥中表达绿色荧光蛋白（GFP）标记的α-微管蛋白 6（GFP-TUA6）的锥形细胞中微管组织的方法。这些活体成像方法可能会使我们对微管在锥形细胞成形中的作用的认识迅速提高。
