Science Research Center, Organization for Research Initiatives, Yamaguchi University, Yamaguchi, Japan.
Department of Horticulture, Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur, Bangladesh.
Methods Mol Biol. 2022;2526:201-213. doi: 10.1007/978-1-0716-2469-2_15.
Responses of plant cells to reactive oxygen species (ROS), e.g., reprogramming of defense genes or progression of cell death, should include the ROS signal transmission to target proteins, but the biochemistry of this process is largely unknown. Lipid peroxide-derived α,β-unsaturated aldehydes and ketones (reactive carbonyl species; RCS), downstream products of ROS stimuli, are recently emerging endogenous agents that can mediate ROS signal to proteins via covalent modification. The involvement of RCS in certain ROS signaling in plants (oxidative injury of leaves and roots, ROS-induced programmed cell death, senescence, and abscisic acid and auxin signaling) has been verified by the determination of RCS with the use of conventional HPLC. Because distinct kinds of RCS act differently in the cell and so are metabolized, identification and quantification of each RCS in plant tissues provide central information to decipher biochemical mechanisms of plant responses to ROS. This article illustrates practical methods of plant sample preparation and extraction and analysis of RCS.
植物细胞对活性氧（ROS）的反应，例如防御基因的重新编程或细胞死亡的进展，应包括 ROS 信号向靶蛋白的传递，但该过程的生物化学在很大程度上是未知的。脂质过氧化物衍生的α，β-不饱和醛和酮（反应性羰基化合物；RCS）是 ROS 刺激的下游产物，是最近出现的内源性试剂，可通过共价修饰将 ROS 信号传递至蛋白质。通过使用常规 HPLC 确定 RCS，已经验证了 RCS 在植物中某些 ROS 信号转导中的作用（叶片和根部的氧化损伤，ROS 诱导的程序性细胞死亡，衰老以及脱落酸和生长素信号转导）。由于不同类型的 RCS 在细胞中作用不同并且被代谢，因此鉴定和定量植物组织中的每种 RCS 可提供有关植物对 ROS 反应的生化机制的关键信息。本文说明了植物样品制备和提取以及 RCS 分析的实用方法。
