Arabidopsis harbors two α and two β genes of pyrophosphate:fructose-6-phosphate 1-phosphotransferase (PFP). The spatial expression patterns of the two AtPFPα genes were analyzed using transgenic plants containing a promoter::ß-glucuronidase (GUS) fusion construct. Whereas the AtPFPα1 promoter was found to be ubiquitously active in all tissues, the AtPFPα2 promoter is preferentially expressed in specific heterotrophic regions of the Arabidopsis plant such as the trichomes of leaves, cotyledon veins, roots, and the stamen and gynoecium of the flowers. Serial deletion analysis of the AtPFPα2 promoter identified a key regulatory element from nucleotides −194 to −175, CGAAAAAGGTAAGGGTATAT, which we have termed PFPα2 and which is essential for AtPFPα2 gene expression. Using a GUS fusion construct driven by this 20-bp sequence in conjunction with a −46 CaMV35S minimal promoter, we also demonstrate that PFPα2 is sufficient for normal AtPFPα2 expression. Hence, this element can not only be used to isolate essential DNA-binding protein(s) that control the expression of the carbon metabolic enzyme AtPFPα2, but has also the potential to be utilized in the production of useful compounds in a specific organ such as the leaf trichomes.