ABSTRACTPlant epidermal cells express unique molecular machinery that juxtapose the assembly of intracellular lipid components and the unique extracellular cuticular lipids that are unidirectionally secreted to plant surfaces. In maize (Zea maysL.), mutations at theglossy2 (gl2)locus affect the deposition of extracellular cuticular lipids. Sequence-based genome scanning identified a novelgl2homolog in the maize genome,Gl2-like. Sequence homology identifies that both theGl2-likeandGl2genes are members of the BAHD superfamily of acyltransferases, with close sequence homology to the ArabidopsisCER2gene. Transgenic experiments demonstrate thatGl2-likeandGl2functionally complement the Arabidopsiscer2mutation, with differential impacts on the cuticular lipids and the lipidome of the plant, particularly affecting the longer alkyl chain acyl lipids, particularly at the 32-carbon chain length. Site-directed mutagenesis of the putative BAHD catalytic HXXXDX-motif indicates thatGl2-likerequires this catalytic capability to fully complement thecer2function, butGl2can accomplish this without the need for this catalytic motif. These findings demonstrate that bothGl2andGl2-likeoverlap in their cuticular lipid function, however the two genes have evolutionary diverged to acquire non-overlapping functions.One-sentence summaryTransgenesis dissection of the functional roles of the maizeGlossy2andGlossy2-Likegenes in cuticular lipid deposition.