REPRISAL: mapping lignification dynamics using chemistry, data segmentation, and ratiometric analysis

• DOI: 10.1093/plphys/kiab490
• Publication Date: 2021-10-20T18:07:02Z
• Authors: Oriane Morel; Cedric Lion; Godfrey Neutelings; Jonathan Stefanov; Fabien Baldacci-C...; Clemence Simon; Christophe Biot; Simon Hawkins; Corentin Spriet
• Keywords: Monolignol; Linum

This article describes a methodology for detailed mapping of the lignification capacity plant cell walls that we have called "REPRISAL" REPorter Ratiometrics Integrating Segmentation Analyzing Lignification. REPRISAL consists combination three separate approaches. In first approach, H*, G*, and S* monolignol chemical reporters, corresponding to p-coumaryl alcohol, coniferyl sinapyl are used label growing lignin polymer in fluorescent triple labeling strategy based on sequential use main bioorthogonal reactions. second step, an automatic parametric and/or artificial intelligence segmentation algorithm is developed assigns image pixels distinct wall zones corners, compound middle lamella secondary walls. The last step corresponds exploitation ratiometric approach enabling statistical analyses differences reporter distribution (ratiometric method [RM] 1) proportions (RM 2) within different zones. We describe this map developmentally related changes wild-type Arabidopsis (Arabidopsis thaliana) interfascicular fiber cells. then apply analyze peroxidase (PRX) mutant prx64 provide further evidence implication AtPRX64 protein floral stem lignification. addition, also demonstrate general applicability by using it poplar (Populus tremula × Populus alba), flax (Linum usitatissimum), maize (Zea mays). Finally, show can be incorporation fucose into noncellulosic polymers.