Changes in Arabidopsis thaliana seedling cell wall assembly induced by treatment with Yariv reagent – Molecular features & visualization with immunocytochemistry and a fluorescent Yariv reagent

Abstract

Arabinogalactan proteins (AGPs) are cell wall-plasma membrane proteins with a high level of glycosylation. The selective and high-affinity binding between AGP and the Yariv reagent has been widely used to carry out functional studies on AGPs by disrupting AGP functions using a non-genetic tool. The current work aimed to determine the molecular features of cell walls during Arabidopsis thaliana seed germination under conditions where AGP functions are blocked. To achieve this, we used molecular & imaging methods with molecular probes and for the first time − a new tool for AGP detection − a fluorescent analogue of the Yariv reagent. The most significant changes included a decrease in the content of AGPs, due to the addition of the Yariv reagent, and subsequent changes only in the content of AGPs upon transfer from the Yariv reagent to fresh Yariv-free medium. Additionally, as a result of the presence of the Yariv reagent, changes in the molecular masses of the analysed cell wall components were observed: lack of AGPs with small molecular mass and disappearance of homogalacturonan with high molecular mass. This work provided the first example of AGP labelling using antibodies and AzYariv-Cy5, and highlights the utility of AzYariv-Cy5 as a broad-spectrum tool for AGP studies.