Yin Xiaofeng, Tsukaya Hirokazu
Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan.
International Research Fellow of Japan Society for the Promotion of Science, Tokyo, Japan.
Bio Protoc. 2021 Jan 5;11(1):e3882. doi: 10.21769/BioProtoc.3882.
In plants, the morphological diversity of leaves is largely determined by cell division, especially cell division orientation. Whereas cell division itself is easily monitored, the detection and quantification of cell division orientation are difficult. The few existing methods for detection and quantification of cell division orientation are either inefficient or laborious. Here, we describe a pulse-chase strategy using a 5-ethynyl-2'-deoxyuridine (EdU) labeling assay. Plant tissues are first incubated with EdU for a short period (pulse), followed by a long incubation without EdU (chase). Using this method, the positions of daughter cells are easily detected and can be used to quantify cell division orientation. Our protocol is rapid and very efficient for quantitative analysis of cell division orientation, and can be applied to both model and non-model plant species. .
在植物中，叶片的形态多样性很大程度上由细胞分裂决定，尤其是细胞分裂方向。虽然细胞分裂本身易于监测，但细胞分裂方向的检测和定量却很困难。现有的少数检测和定量细胞分裂方向的方法要么效率低下，要么费力。在这里，我们描述了一种使用5-乙炔基-2'-脱氧尿苷（EdU）标记测定的脉冲追踪策略。首先将植物组织与EdU短时间孵育（脉冲），然后在无EdU的情况下长时间孵育（追踪）。使用这种方法，可以轻松检测子细胞的位置，并可用于量化细胞分裂方向。我们的方案对于细胞分裂方向的定量分析快速且非常有效，并且可以应用于模式植物和非模式植物物种。
