A Pulse–chase EdU Method for Detection of Cell Division Orientation in Arabidopsis and Juncus prismatocarpus Leaf Primordia

Abstract

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 pulsechase 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.

Biregional division angles generate sharp apex and concave joints in leaves

“…The protocol for the detection of cell division angles using EdU pulse-chase was modified from that of Yin and Tsukaya (2021). Leaf primordia of T. sebifera were collected from seedlings and saplings.…”