title: BLISS: A Bioorthogonal Dual-Labeling Strategy to Unravel Lignification Dynamics in Plants.
abstract: A better in vivo understanding of lignin formation within plant cell walls will contribute to improving the valorization of plant-derived biomass. Although bioorthogonal chemistry provides a promising platform to study the lignification process, methodologies that simultaneously detect multiple chemical reporters in living organisms are still scarce. Here, we have developed an original bioorthogonal labeling imaging sequential strategy (BLISS) to visualize and analyze the incorporation of both p-hydroxyphenyl (H) and guaiacyl (G) units into lignin in vivo with a combination of strain-promoted and copper-catalyzed azide-alkyne cycloadditions. On our path to BLISS, we designed a new azide-tagged monolignol reporter for H units in metabolic lignin engineering and used it in conjunction with an alkyne-tagged G unit surrogate to study lignification dynamics in flax. Here, we show that BLISS provides precise spatial information on the zones of active lignification and reveals polarization in single-cell lignification dynamics.
authors.affiliation: Université de Lille, CNRS, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, 59000 Lille, France. Electronic address: cedric.lion@univ-lille1.fr.
authors.forename: Cedric
authors.initials: C
authors.lastname: Lion
authors.affiliation: Université de Lille, CNRS, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, 59000 Lille, France.
authors.forename: Clémence
authors.initials: C
authors.lastname: Simon
authors.affiliation: Université de Lille, CNRS, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, 59000 Lille, France.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Huss
authors.affiliation: Université de Lille, CNRS, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, 59000 Lille, France.
authors.forename: Anne-Sophie
authors.initials: AS
authors.lastname: Blervacq
authors.affiliation: Université de Lille, CNRS, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, 59000 Lille, France.
authors.forename: Louis
authors.initials: L
authors.lastname: Tirot
authors.affiliation: Université de Lille, CNRS, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, 59000 Lille, France.
authors.forename: Djadidi
authors.initials: D
authors.lastname: Toybou
authors.affiliation: Université de Lille, CNRS, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, 59000 Lille, France.
authors.forename: Corentin
authors.initials: C
authors.lastname: Spriet
authors.affiliation: Université de Lille, Inserm, U1003 - PHYCEL - Physiologie Cellulaire, 59000 Lille, France.
authors.forename: Christian
authors.initials: C
authors.lastname: Slomianny
authors.affiliation: Université de Lille, CNRS, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, 59000 Lille, France.
authors.forename: Yann
authors.initials: Y
authors.lastname: Guerardel
authors.affiliation: Université de Lille, CNRS, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, 59000 Lille, France. Electronic address: simon.hawkins@univ-lille1.fr.
authors.forename: Simon
authors.initials: S
authors.lastname: Hawkins
authors.affiliation: Université de Lille, CNRS, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, 59000 Lille, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Biot
mesh_terms: D000480:Alkynes / Q000737:chemistry; D001386:Azides / Q000737:chemistry; D002384:Catalysis; D002473:Cell Wall / Q000737:chemistry / Q000378:metabolism; D003300:Copper / Q000737:chemistry; D003373:Coumaric Acids; D061565:Cycloaddition Reaction; D019597:Flax / Q000737:chemistry / Q000378:metabolism; D008031:Lignin / Q000737:chemistry* / Q000378:metabolism; D008856:Microscopy, Fluorescence; D010944:Plants / Q000737:chemistry / Q000378:metabolism*; D011422:Propionates / Q000737:chemistry; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D000480:Alkynes; D001386:Azides; D003373:Coumaric Acids; D011422:Propionates; D003300:Copper; D008031:Lignin; C495469:p-coumaric acid
keywords: CuAAC; SPAAC; bioorthogonal chemistry; click chemistry; dual labeling; lignification; lignin; metabolic labeling; monolignol; plant cell wall