CarboTag: a modular approach for live and functional imaging of plant cell walls
Maarten Besten, Milan Hendriksz, Lucile Michels, Bénédicte Charrier, Elwira Smakowska‐Luzan, Dolf Weijers, Jan Willem Borst, Joris Sprakel
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. A suite of nontoxic and permeable CarboTag probes in various colors enables live quantitative imaging of a range of plant cell wall characteristics and dynamic, high-resolution mapping of cell wall changes in response to growth or perturbations.