Visualizing Lignification Dynamics in Plants with Click Chemistry: Dual Labeling is BLISS!
Open Access
- 26 January 2018
- journal article
- research article
- Published by MyJove Corporation in Journal of Visualized Experiments
- No. 131,p. e56947
- https://doi.org/10.3791/56947
Abstract
Lignin is one of the most prevalent biopolymers on the planet and a major component of lignocellulosic biomass. This phenolic polymer plays a vital structural and protective role in the development and life of higher plants. Although the intricate mechanisms regulating lignification processes in vivo strongly impact the industrial valorization of many plant-derived products, the scientific community still has a long way to go to decipher them. In a simple three-step workflow, the dual labeling protocol presented herein enables bioimaging studies of actively lignifying zones of plant tissues. The first step consists in the metabolic incorporation of two independent chemical reporters, surrogates of the two native monolignols that give rise to lignin H- and G-units. After incorporation into growing lignin polymers, each reporter is then specifically labeled with its own fluorescent probe via a sequential combination of bioorthogonal SPAAC/CuAAC click reactions. Combined with lignin autofluorescence, this approach leads to the generation of three-color localization maps of lignin within plant cell walls by confocal fluorescence microscopy and provides precise spatial information on the presence or absence of active lignification machinery at the scale of plant tissues, cells and different cell wall layers.This publication has 28 references indexed in Scilit:
- Bifunctional Unnatural Sialic Acids for Dual Metabolic Labeling of Cell-Surface Sialylated GlycansJournal of the American Chemical Society, 2013
- Two‐Color Glycan Labeling of Live Cells by a Combination of Diels–Alder and Click ChemistryAngewandte Chemie, 2013
- Natural Hypolignification Is Associated with Extensive Oligolignol Accumulation in Flax StemsPlant Physiology, 2012
- Metabolic click-labeling with a fucose analog reveals pectin delivery, architecture, and dynamics in Arabidopsis cell wallsProceedings of the National Academy of Sciences of the United States of America, 2012
- Structural Characterization of Guaiacyl-rich Lignins in Flax (Linum usitatissimum) Fibers and ShivesJournal of Agricultural and Food Chemistry, 2011
- The origin and evolution of lignin biosynthesisNew Phytologist, 2010
- Bioorthogonal Chemistry: Fishing for Selectivity in a Sea of FunctionalityAngewandte Chemie, 2009
- Metabolic Labeling of Sialic Acids in Living Animals with Alkynyl SugarsAngewandte Chemie, 2009
- Chemistry in living systemsNature Chemical Biology, 2005
- Lignin BiosynthesisAnnual Review of Plant Biology, 2003