Iron (Fe) deficiency is a world-wide serious agricultural problem. Maize secretes 2'-deoxymugineic acid (DMA) to uptake and utilize Fe from the soil. In order to explore the gene expression patterns of the DMA secretion channel gene YS3, we cloned the 2813 bp YS3 promoter, and constructed the plant expression vector pCAMBIA-YS3GUS. The promoter contains a lot of TATA-boxes and CAAT-boxes, and cis-acting regulatory elements such as the light responsive elements and the hormone responsive elements. Arabidopsis was transformed via Agrobacterium tumefacients-mediated procedures to obtain the pYS3::GUS transgenic plants, which were confirmed by GUS staining. Then, the stained tissue was observed using paraffin section methods and the YS3 promoter activity was also analyzed. We found that the promoter could drive GUS gene expression specifically in the root epidermal cells. Mechanical damage could activate the promoter, and drive the GUS gene expression adjacent to the damage sites. Our results provide a molecular basis to understand the DMA secretion process in maize.