Albert Markus, Fürst Ursula
Center for Plant Molecular Biology, University of Tübingen, Auf der Morgenstelle 32, 72076, Tübingen, Germany.
Methods Mol Biol. 2017;1621:69-76. doi: 10.1007/978-1-4939-7063-6_7.
The oxidative burst or the production of reactive oxygen species (ROS) is a typical cellular response of both plants and animals to diverse abiotic and biotic stresses. Mainly, the (re-)active oxygen species include the superoxide anion (O), hydrogen peroxide (HO), and the hydroxyl radical (OH•). Here, we outline the detection of extracellularly produced ROS in plant leaf pieces using a chemiluminescence-based bioassay with the luminol L-012 as a substrate being oxidized in the presence of ROS. Since this type of assay is in use in many laboratories, e.g., as a readout for activation of plant receptor kinases, we include a discussion on the interpretation of results and points addressing problems with the buffers at suboptimal pH values that negatively influence the chemiluminescence production.
氧化爆发或活性氧（ROS）的产生是动植物对各种非生物和生物胁迫的典型细胞反应。主要地，（活性）氧包括超氧阴离子（O）、过氧化氢（HO）和羟基自由基（OH•）。在此，我们概述了使用基于化学发光的生物测定法检测植物叶片碎片中细胞外产生的ROS，该测定法以鲁米诺L-012作为底物，在ROS存在下被氧化。由于这种类型的测定法在许多实验室中都有使用，例如作为植物受体激酶激活的读数，我们讨论了结果的解释以及关于在非最佳pH值下对化学发光产生有负面影响的缓冲液问题。
