2016 journal article

A flow cytometric method for estimating S-phase duration in plants

JOURNAL OF EXPERIMENTAL BOTANY, 67(21), 6077–6087.

By: L. Mickelson-Young n, E. Wear n, P. Mulvaney n, T. Lee n, E. Szymanski  n, G. Allen n, L. Hanley-Bowdoin n , W. Thompson n 

author keywords: Arabidopsis; barley; EdU; flow cytometry; maize; replication timing; rice; S-phase duration; wheat
MeSH headings : Arabidopsis / cytology; Arabidopsis / growth & development; DNA, Plant / metabolism; Deoxyuridine / analogs & derivatives; Deoxyuridine / metabolism; Flow Cytometry / methods; G1 Phase / physiology; G2 Phase / physiology; Hordeum / cytology; Hordeum / growth & development; Meristem / cytology; Meristem / growth & development; Oryza / cytology; Oryza / growth & development; S Phase / physiology; Triticum / cytology; Triticum / growth & development; Zea mays / cytology; Zea mays / growth & development
topics (OpenAlex): Chromosomal and Genetic Variations; Plant nutrient uptake and metabolism; Plant Genetic and Mutation Studies
TL;DR: This work estimated S-phase duration for several plant species by following EdU-labeled nuclei from G1 to G2 using bivariate flow cytometry, finding that S- phase duration is relatively consistent over a range of genome sizes. (via Semantic Scholar)
UN Sustainable Development Goals Color Wheel
UN Sustainable Development Goal Categories
2. Zero Hunger (Web of Science)
13. Climate Action (Web of Science)
Source: Web Of Science
Added: August 6, 2018

The duration of the DNA synthesis stage (S phase) of the cell cycle is fundamental in our understanding of cell cycle kinetics, cell proliferation, and DNA replication timing programs. Most S-phase duration estimates that exist for plants are based on indirect measurements. We present a method for directly estimating S-phase duration by pulse-labeling root tips or actively dividing suspension cells with the halogenated thymidine analog 5-ethynl-2'-deoxyuridine (EdU) and analyzing the time course of replication with bivariate flow cytometry. The transition between G1 and G2 DNA contents can be followed by measuring the mean DNA content of EdU-labeled S-phase nuclei as a function of time after the labeling pulse. We applied this technique to intact root tips of maize (Zea mays L.), rice (Oryza sativa L.), barley (Hordeum vulgare L.), and wheat (Triticum aestivum L.), and to actively dividing cell cultures of Arabidopsis (Arabidopsis thaliana (L.) Heynh.) and rice. Estimates of S-phase duration in root tips were remarkably consistent, varying only by ~3-fold, although the genome sizes of the species analyzed varied >40-fold.