ZmEREB46, a maize ortholog of Arabidopsis WAX INDUCER1/SHINE1, is involved in the biosynthesis of leaf epicuticular very-long-chain waxes and drought tolerance.

The aerial surfaces of plants are covered by a layer of cuticular wax that is composed of long-chain hydrocarbon compounds for protection against adverse environmental conditions. The current study identified a maize (Zea mays L.) APETALA2/ethylene-responsive element-binding protein (AP2/EREBP)-type transcription factor, ZmEREB46. Ectopic expression of ZmEREB46 in Arabidopsis increased the accumulation of epicuticular wax on the leaves and enhanced the drought tolerance of plants. The amounts of C24/C32 fatty acids, C32/C34 aldehydes, C32/C34 1-alcohols and C31 alkanes in zmereb46 (ZmEREB46 knockout mutant) leaves were reduced. The amount of leaf total epicuticular wax decreased approximately 50% in zmereb46. Compared to wild-type LH244 leaves, the cuticle permeability of zmereb46 leaves was increased, which resulted from decreased epicuticular wax load and a thinner cuticle layer. ZmEREB46 had transcriptional activation activity and directly bound to promoter regions of ZmCER2, ZmCER3.2 and ZmKCS1. The zmereb46 seedlings also exhibited reduced drought tolerance. These results, and the observations in ZmEREB46-overexpressing lines, suggest that ZmEREB46 is involved in cuticular metabolism by influencing the biosynthesis of very-long-chain waxes and participates in the cutin biosynthesis pathway. These results are helpful to further analyze the regulatory network of wax accumulation in maize.