CarboTag: a modular approach for live and functional imaging of plant cell walls.

Besten, Maarten; Hendriksz, Milan; Michels, Lucile; Charrier, Bénédicte; Smakowska-Luzan, Elwira; Weijers, Dolf; Borst, Jan Willem; Sprakel, Joris · Nat Methods · 2025

basic_science · Level V

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Abstract

Plant cells are contained within a rigid network of cell walls. Cell walls serve as a structural material and a crucial signaling hub vital to all aspects of the plant life cycle. However, many features of the cell wall remain enigmatic, as it has been challenging to map its functional properties in live plants at subcellular resolution. Here, we introduce CarboTag, a modular toolbox for live functional imaging of plant walls. CarboTag uses a small molecular motif, a pyridine boronic acid, that directs its cargo to the cell wall. We designed a suite of cell wall imaging probes based on CarboTag in various colors for multiplexing. Additionally, we developed new functional reporters for live quantitative imaging of key cell wall characteristics: network porosity, cell wall pH and the presence of reactive oxygen species. CarboTag paves the way for dynamic and quantitative mapping of cell wall responses at subcellular resolution.

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