Abstract

Xylan, a pivotal polymer with diversified structures, is indispensable for cell wall integrity and contributes to plant growth and biomass recalcitrance. Xylan is synthesized by multienzyme complexes named xylan synthase complexes (XSCs). However, the biochemical mechanism of XSCs and the functions of core components within XSC remain unclear. Here, we report that rice (Oryza sativa) XYLAN O-ACETYLTRANSFERASE 6 (XOAT6) and the xylan synthase IRREGULAR XYLEM10 (IRX10) represent core components of the XSC, acting together to biosynthesize acetyl-xylans. Co-fractionation mass spectrometry and protein–protein interaction analyses revealed that IRX10 and XOAT6 physically interact within XSC, corroborated by similar xylan defects in xoat6 and irx10 mutants. Biochemical assays showed that XOAT6 is an O-acetyltransferase of the xylan backbone and facilitates chain polymerization catalyzed by IRX10. Fluorescence correlation spectroscopy further visualized the xylooligomer polymerization process at a single-molecule level. Solid-state NMR analysis, electron microscopy observations, and nanoindentation examinations identified the altered xylan conformation, disorganized cellulosic structure, and increased wall rigidity and cellulose accessibility in the mutants, leading to brittleness and improved saccharification efficiency. Our findings provide insights into the assembly of XSCs and xylan biosynthesis and offer a framework for tailoring xylans to improve crop traits and biomass.

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