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Early radiolabeling experiments performed by Kolattukudy’s group 30 years agogave insight into the pathways involved in oxidation of C 16 and C 18 monomers, using cutinizing (. extracts of bean leaf epidermis) and suberizing (. wound-healed potato slices) tissues. Since enzymes involved in biosynthesis of cutin/suberin monomers are possibly associated to membranes and/or are part of enzyme complexes, biochemical approaches to isolate these proteins have not been successful. Several recombinant plant fatty acid ω-hydroxylases were shown to be active in yeast expression systems, but these experiments could not demonstrate their involvement in cutin or suberin. Thus, genetic approaches have been particularly helpful to dissect biosynthetic pathways. Several cutin mutants have been identified in part because they resemble transgenic plants that overexpress fungal cutinase (cutinases are enzymes that hydrolyze the ester linkages), which show alterations in the structure of the cuticle and organ fusions, or because they present increased susceptibility to pathogens. Further development of quantitative analyses of lipid polyester monomers in Arabidopsis allowed the use of reverse genetic approaches to discover new genes involved in lipid polyester synthesis. Identification of candidate genes has been aided by published transcriptome analyses from Arabidopsis epidermis and cork tree epidermis, as well as public repositories for microarray data. Reverse genetics has also been recently applied to demonstrate the function of suberin-synthesizing genes in potato periderm.