Monolignols have important roles in plant development and primarily serve as monomers for lignin polymerization in secondary cell walls. Monolignols are synthesized in the plant cytoplasm prior to entering the apoplast where oxidation occurs. Upon oxidation, monolignols are incorporated into the cell wall. Several mechanisms have been suggested to explain how monolignols cross the plasma membrane, including endocytosis, diffusion, and active transport. However, evidence for those models relied on theoretical calculations or produced results using in vitro approaches. Further, only one active transporter protein has been characterized to date. Yet, of the three monolignols tested, the transporter was only demonstrably shown to transport p-coumaryl alcohol, the least abundant monolignol present in Arabidopsis.Here we show that AtPMT4 is likely a monolignol transporter, particularly for the more abundant monolignols: coniferyl alcohol and sinapyl alcohol. Gene expression analysis performed on AtPMT4 in dicots and monocots shows coexpression with lignin biosynthetic genes. Cell-specific expression analysis of the inflorescence stem, a tissue that undergoes intense lignification to provide plant structural support, shows that AtPMT4 is expressed higher in cell types that lignify. We demonstrate that Arabidopsis Col-0 plants transformed with a CRISPR-Cas9 construct targeted near the TSS of AtPMT4, a member of the POLYOL/ MONOSACCHARIDE TRANSPORTER family, which is a subfamily of the MONOSACCHARIDE TRANSPORTER-LIKE family, displayed altered lignin phenotypes. We quantified the total lignin, free monomer subunits, and digestibility of the inflorescence stem in pmt4. Our studies show lower amounts of lignin and increased digestibility when AtPMT4 is mutated. Further, we show that pmt4 is sensitive to monolignols when grown in the presence of coniferyl alcohol. pmt4 displayed shorter root length compared to Col-0 at low concentrations of coniferyl alcohol. In conclusion, we provide evidence for an understanding of monolignol translocation and lignification by which transporters are likely involved in a proton-coupled manner.
Read
