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A simple pipeline for cell cycle kinetic studies in the root apical meristem. | LitMetric

AI Article Synopsis

  • The study focuses on how root system architecture is influenced by communication between cells in the root apical meristem (RAM) and environmental factors.
  • Researchers developed a method to analyze cell parameters and DNA replication in the RAM without using specific markers, employing techniques like EdU staining.
  • Differences were observed in cell and nucleus volumes, and DNA replication timing varied by cell type, with the G2 phase lasting longer in the epidermis compared to the pericycle, highlighting the method's potential for studying cell cycle variations in different plant species.

Article Abstract

Root system architecture ultimately depends on precise signaling between different cells and tissues in the root apical meristem (RAM) and integration with environmental cues. This study describes a simple pipeline to simultaneously determine cellular parameters, nucleus geometry, and cell cycle kinetics in the RAM. The method uses marker-free techniques for nucleus and cell boundary detection, and 5-ethynyl-2'-deoxyuridine (EdU) staining for DNA replication quantification. Based on this approach, we characterized differences in cell volume, nucleus volume, and nucleus shape across different domains of the Arabidopsis RAM. We found that DNA replication patterns were cell layer and region dependent. G2 phase duration, which varied from 3.5 h in the pericycle to more than 4.5 h in the epidermis, was found to be associated with some features of nucleus geometry. Endocycle duration was determined as the time required to achieve 100% EdU-positive cells in the elongation zone and, as such, it was estimated to be in the region of 5 h for the epidermis and cortex. This experimental pipeline could be used to precisely map cell cycle duration in the RAM of mutants and in response to environmental stress in several plant species without the need for introgressing molecular cell cycle markers.

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Source
http://dx.doi.org/10.1093/jxb/erac123DOI Listing

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