Université de Lille, CNRS, UMR 8576, UGSF-, Unité de Glycobiologie Structurale et Fonctionnelle, 59000, Lille, France.
Angew Chem Int Ed Engl. 2018 Dec 17;57(51):16665-16671. doi: 10.1002/anie.201808493. Epub 2018 Nov 21.
Reported herein is an in vivo triple labelling strategy to monitor the formation of plant cell walls. Based on a combination of copper-catalysed alkyne-azide cycloaddition (CuAAC), strain-promoted azide-alkyne cycloaddition (SPAAC), and Diels-Alder reaction with inverse electronic demand (DAR ), this methodology can be applied to various plant species of interest in research. It allowed detection of the differential incorporation of alkynyl-, azido-, and methylcyclopropenyl-tagged reporters of the three main monolignols into de novo biosynthesized lignin in different tissues, cell types, or cell wall layers. In addition, this triple labelling was implemented with different classes of chemical reporters, using two monolignol reporters in conjunction with alkynylfucose to simultaneously monitor the biosynthesis of lignin and non-cellulosic polysaccharides. This allowed observation of their deposition occurring contemporaneously in the same cell wall.
本文报道了一种用于监测植物细胞壁形成的体内三重标记策略。该策略基于铜催化的叠氮-炔环加成（CuAAC）、应变促进的叠氮-炔环加成（SPAAC）和逆电子需求的 Diels-Alder 反应（DAR）的组合，可应用于各种具有研究意义的植物物种。该方法可用于检测三种主要木质素单体的炔基、叠氮基和甲基环丙烯基标记物在不同组织、细胞类型或细胞壁层中从头生物合成木质素的差异掺入。此外，还使用不同类别的化学报告物实现了三重标记，同时使用两种木质素报告物和炔基岩藻糖来监测木质素和非纤维素多糖的生物合成。这使得可以观察到它们同时在同一细胞壁中沉积。
