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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. [ABSTRACT FROM AUTHOR]

To protect above‐ground plant organs from excessive water loss, their surfaces are coated by waxes. The genes involved in wax formation have been investigated in detail in Arabidopsis but scarcely in crop species. Here, we aimed to isolate and characterize a CER1 enzyme responsible for formation of the very long‐chain alkanes present in high concentrations especially during late stages of wheat development. On the basis of comparative wax and transcriptome analyses of various wheat organs, we selected TaCER1‐1A as a primary candidate and demonstrated that it was located to the endoplasmic reticulum, the subcellular compartment for wax biosynthesis. A wheat nullisomic‐tetrasomic substitution line lacking TaCER1‐1A had significantly reduced amounts of C33 alkane, whereas rice plants overexpressing TaCER1‐1A showed substantial increases of C25–C33 alkanes relative to wild type control. Similarly, heterologous expression of TaCER1‐1A in Arabidopsis wild type and the cer1 mutant resulted in increased levels of unbranched alkanes, iso‐branched alkanes and alkenes. Finally, the expression of TaCER1‐1A was found activated by abiotic stresses and abscisic acid treatment, resulting in increased production of alkanes in wheat. Taken together, our results demonstrate that TaCER1‐1A plays an important role in wheat wax alkane biosynthesis and involved in responding to drought and other environmental stresses. In this paper, the function of TaCER1‐1A involved in wheat alkane biosynthesis was tested using gain‐of‐function (transgenic plants) and loss‐of‐function plants (gene deletion wheat line), and the role of alkanes in drought resistance was assessed. [ABSTRACT FROM AUTHOR]

Plant cuticular wax is a heterogeneous mixture of very long chain fatty acids (VLCFAs) and their derivatives. Primary alcohols are the dominant wax components throughout leaf development of Brachypodium distachyon (Brachypodium). However, the genes involved in primary alcohol biosynthesis have not been investigated and their exact biological function remains unclear in Brachypodium to date. Here, we monitored the leaf wax profile and crystal morphology during Brachypodium leaf morphogenesis, and isolated three Brachypodium fatty acyl-CoA reductase (FAR) genes, named BdFAR1, BdFAR2 and BdFAR3, then analyzed their biochemical activities, substrate specificities, expression patterns, subcellular localization and stress induction. Transgenic expression of BdFAR genes in yeast (Saccharomyces cerevisiae), tomato (Solanum lycopersicum), Arabidopsis (Arabidopsis thaliana) and Brachypodium increased the production of primary alcohols. The three BdFAR genes were preferentially expressed in Brachypodium aerial tissues, consistent with known sites of wax primary alcohol deposition, and localized in the endoplasmic reticulum (ER) in Arabidopsis protoplasts. Finally, expression of the BdFAR genes was induced by drought, cold and ABA treatments, and drought stress significantly increased cuticular wax accumulation in Brachypodium. Taken together, these results indicate that the three BdFAR genes encode active FARs involved in the biosynthesis of Brachypodium wax primary alcohols and respond to abiotic stresses. [ABSTRACT FROM AUTHOR]