Arabidopsis thaliana NIP5;1, a boric acid diffusion facilitator, is involved in the acquisition of boron (B) from soil for growth under B limitation. AtNIP5;1 is expressed mainly in roots, where its expression is highest in the root cap and elongation zone. Here, we studied the role of the AtNIP5;1 promoter in the expression of this gene in roots. We fused a series of AtNIP5;1 promoter variants with deleted 5'-fragments to the GUS reporter gene and investigated the expression patterns by histochemical staining. We found that three regions of the AtNIP5;1 promoter are required for specific expression in the root cap and elongation zone (-880 to -863 bp from the translation start site), distal side of the differentiation zone (-747 to -722 bp), and basal side of the differentiation zone (-661 and -621 bp). The results suggest that at least three regions of the AtNIP5;1 promoter each confer different cell-type-specific expression.