Foyer Christine H, Pellny Till K, Locato Vittoria, Hull Jonathon, De Gara Laura
Faculty of Biological Sciences, University of Leeds, Leeds, UK.
Faculty Health and Applied Sciences, University of the West of England, Bristol, UK.
Methods Mol Biol. 2019;1990:165-181. doi: 10.1007/978-1-4939-9463-2_14.
Reactive oxygen species (ROS) and low molecular weight antioxidants, such as glutathione and ascorbate, are powerful signalling molecules that participate in the control of plant growth and development, and modulate progression through the mitotic cell cycle. Enhanced ROS 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 (PARP) 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.
活性氧(ROS)和低分子量抗氧化剂,如谷胱甘肽和抗坏血酸,是强大的信号分子,参与植物生长和发育的调控,并调节有丝分裂细胞周期的进程。ROS积累增强或抗坏血酸或谷胱甘肽水平降低会导致细胞周期停滞,尤其是通过G1检查点时会阻止进程。植物细胞悬浮培养已被证明是研究细胞周期调控的特别有用的工具。在这里,我们提供了有效且准确的方法,用于测量有丝分裂和细胞扩张过程中,特别是在细胞悬浮培养中,细胞内抗坏血酸和谷胱甘肽库的变化以及相关酶如聚(ADP-核糖)聚合酶(PARP)的活性。这些方法可用于旨在增进当前对氧化还原控制在细胞分裂和细胞扩张中作用理解的研究。