Umeå Plant Science Centre, Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences, Umeå, Sweden.
Methods Mol Biol. 2024;2722:139-148. doi: 10.1007/978-1-0716-3477-6_11.
Peroxidases (PRXs) and laccases (LACs) are enzymes involved in catalyzing the oxidation of the lignin monomers to facilitate lignin polymerization. However, due to the large number of genes composing these two families of enzymes, many details regarding their specific localization are only partially understood. Here, we present a fast and easy histochemical method that makes use of the artificial substrate 3,3',5,5'-tetramethylbenzidine (TMB) to visualize PRX and LAC activities in the hybrid aspen (Populus tremula x P. tremuloides) xylem tissue. In addition, we describe a protocol that allows the detection of the PRX substrate, HO, using the nonfluorescent dye 2',7'-dichlorodihydrofluorescein diacetate (HDCFDA) in woody tissues.
过氧化物酶 (PRXs) 和漆酶 (LACs) 是参与催化木质素单体氧化以促进木质素聚合的酶。然而，由于这两种酶家族的基因数量众多，因此对于它们的具体定位只有部分了解。在这里，我们提出了一种快速简便的组织化学方法，该方法利用人工底物 3,3',5,5'-四甲基联苯胺 (TMB) 来可视化杂种白杨 (Populus tremula x P. tremuloides) 木质部组织中的 PRX 和 LAC 活性。此外，我们还描述了一种使用非荧光染料 2',7'-二氯二氢荧光素二乙酸酯 (HDCFDA) 在木质组织中检测 PRX 底物 HO 的方案。
