Analysis of Phragmoplast Kinetics During Plant Cytokinesis

Methods Mol Biol. 2017:1662:137-150. doi: 10.1007/978-1-4939-7262-3_12.

Abstract

In plants, the partitioning of daughter cells during cytokinesis is achieved via physical insertion of a membranous cell plate within the dividing parent cell. It is a cellular process of extensive protein secretion and membrane trafficking toward the plane of cell division and the cytoskeleton is an important facilitator of this process. A specialized cytoskeletal array termed phragmoplast expands centrifugally throughout cytokinesis and directs, mostly Golgi-derived vesicles that ultimately fuse to form the developing cell plate. The function of the phragmoplast in guiding cell plate synthesis has strongly motivated many scientists to monitor its dynamic behavior. In this chapter, we present an overview of basic principles and methods concerning the live imaging of cytokinetic plant cells using confocal laser scanning microscopy (CLSM) and the analysis of phragmoplast expansion.

Keywords: Arabidopsis; CLSM; Cell plate; Cytokinesis; FM4-64; GFP; MAP4; MBD; Microtubules; Phragmoplast.

Publication types

  • Review

MeSH terms

  • Arabidopsis / genetics
  • Arabidopsis / metabolism
  • Arabidopsis / ultrastructure*
  • Cytokinesis*
  • Cytoplasm / metabolism
  • Cytoplasm / ultrastructure
  • Cytoskeleton / metabolism
  • Cytoskeleton / ultrastructure*
  • Fluorescent Dyes / chemistry
  • Gene Expression
  • Genes, Reporter
  • Golgi Apparatus / metabolism
  • Golgi Apparatus / ultrastructure*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Kinetics
  • Microscopy, Confocal / methods
  • Microtubules / metabolism
  • Microtubules / ultrastructure
  • Plant Cells / metabolism
  • Plant Cells / ultrastructure*
  • Plants, Genetically Modified
  • Protein Transport
  • Pyridinium Compounds / chemistry
  • Quaternary Ammonium Compounds / chemistry
  • Transport Vesicles / metabolism
  • Transport Vesicles / ultrastructure*

Substances

  • FM 4-64
  • Fluorescent Dyes
  • N-(3-trimethylammoniumpropyl)-4-(6-(4-(diethylamino)phenyl)hexatrienyl)pyridinium
  • Pyridinium Compounds
  • Quaternary Ammonium Compounds
  • Green Fluorescent Proteins