Induced accumulation of cuticular waxes enhances drought tolerance in Arabidopsis by changes in development of stomata.

Cuticular waxes are involved in the regulation of the exchange of gases and water in plants and can impact tolerance to drought. However, the molecular mechanisms of the relationship between wax accumulation and drought tolerance are largely unknown. We applied the methoxyfenozide gene switching system to regulate expression of the WIN1/SHN1 gene (WAX INDUCER 1/SHINE1; At1G15360), a transcriptional activator, to regulate production of cuticular waxes and cutin and followed changes of gene expression, metabolites, and drought tolerance. Treatment with the inducer resulted in expression of the target gene and specific downstream genes, and gradually increased cuticular waxes. Induction of cuticular wax conferred tolerance to drought and recovery from drought, and was correlated with reduced numbers of stomata. Quantitative RT-PCR assays using RNAs from transgenic plants revealed that when expression of the WIN1/SHN1 gene was induced there was increased expression of genes involved in wax development, and reduced expression of selected genes, including SPCH (At5g53210); MUTE (At3g06120); and FAMA (At3g241400); and YODA (At1g63700), each of which is involved in stomatal development. These studies suggest that drought tolerance caused by the induction of WIN1/SHIN gene may be due to reduced numbers of stomata as well as to cuticular wax accumulation.