Characterization of an alkylresorcinol synthase that forms phenolics accumulating in the cuticular wax on various organs of rye (Secale cereale)

Summary Alkylresorcinols are bioactive compounds produced in diverse plant species, with chemical structures combining an aliphatic hydrocarbon chain and an aromatic ring with characteristic hydroxyl substituents. Here, we aimed to isolate and characterize the enzyme that forms the alkylresorcinols accumulating in the cuticular wax on the surface of all above‐ground organs of rye. Based on sequence homology with other type‐III polyketide synthases, a candidate alkylresorcinol synthase was cloned. Yeast heterologous expression showed that the enzyme, ScARS, is highly specific for the formation of the aromatic resorcinol ring structure, through aldol condensation analogous to stilbene synthases. The enzyme accepts long‐chain and very‐long‐chain acyl‐CoA starter substrates, preferring saturated over unsaturated chains. It typically carries out three rounds of condensation with malonyl‐CoA prior to cyclization, with only very minor activity for a fourth round of malonyl‐CoA condensation and cyclization to 5‐(2′‐oxo)‐alkylresorcinols or 5‐(2′‐hydroxy)‐alkylresorcinols. Like other enzymes involved in cuticle formation, ScARS is localized to the endoplasmic reticulum. ScARS expression patterns were found correlated with alkylresorcinol accumulation during leaf development and across different rye organs. Overall, our results thus suggest that ScARS synthesizes the cuticular alkylresorcinols found on diverse rye organ surfaces. Significance Statement This work identified and characterized the key enzyme forming alkylresorcinols, compounds with mixed aliphatic/phenolic structures accumulating in the cuticular waxes lining all above‐ground organs of the plant.