Analysis of redox relationships in the plant cell cycle: determinations of ascorbate, glutathione and poly (ADPribose)polymerase (PARP) in plant cell cultures

Methods Mol Biol. 2008;476:199-215. doi: 10.1007/978-1-59745-129-1_14.

Abstract

Reactive oxygen species (ROS) and low molecular weight antioxidants, such as glutathione and ascorbate, are powerful signaling molecules that participate in the control of plant growth and development, and modulate progression through the mitotic cell cycle. Enhanced reactive oxygen species accumulation or low levels of ascorbate or glutathione cause the cell cycle to arrest and halt progression especially through the G1 checkpoint. Plant cell suspension cultures have proved to be particularly useful tools for the study of cell cycle regulation. Here we provide effective and accurate methods for the measurement of changes in the cellular ascorbate and glutathione pools and the activities of related enzymes such poly (ADP-ribose) polymerase during mitosis and cell expansion, particularly in cell suspension cultures. These methods can be used in studies seeking to improve current understanding of the roles of redox controls on cell division and cell expansion.

MeSH terms

  • Arabidopsis / cytology*
  • Arabidopsis / enzymology*
  • Ascorbic Acid / analysis*
  • Cell Cycle*
  • Glutathione / analysis*
  • Molecular Biology / methods*
  • Oxidation-Reduction
  • Poly(ADP-ribose) Polymerases / metabolism*

Substances

  • Poly(ADP-ribose) Polymerases
  • Glutathione
  • Ascorbic Acid