Elucidating the Roles of Three β-Glucuronosyltransferases Acting on Arabinogalactan-Proteins Using a CRISPR-Cas9 Multiplexing Approach in Arabidopsis

In the 2020 study by Zhang et al., the roles of three β-glucuronosyltransferases (GLCAT14A, GLCAT14B, and GLCAT14C) in Arabidopsis were explored using CRISPR-Cas9 gene editing to create single, double, and triple mutants. These enzymes are crucial for attaching glucuronic acid to arabinogalactan-proteins (AGPs), which are important for plant biological processes. The research found that mutants, especially the double and triple mutants, had altered AGP content, monosaccharide composition with less glucuronic acid, and decreased calcium binding. These changes led to various developmental issues, including delayed seed germination, shorter root hair length, less trichome branching, defective pollen grains, shorter siliques, reduced seed set, and less adherent seed coat mucilage. The study concluded that GLCAT14A-C are significant in transferring glucuronic acid to AGPs, affecting multiple biochemical and physiological plant phenotypes.