The Opaque 2 (O2) gene encodes a transcriptional activator of the basic region/leucine zipper family, which controls the synthesis of a major storage protein class in maize endosperm, the 22 kDa alpha-zeins, and of several other non-zein polypeptides including b32. We demonstrate, by analysing O2 mRNAs in different organs of maize plants, that the O2 gene is only active in the endosperm. Its transcription is precisely controlled during seed development: O2 mRNAs are first detected 10 days after pollination and accumulate in the endosperm over a period of 20 days. When introduced into tobacco plants, the O2 promoter directs the expression of the beta-glucuronidase (GUS) reporter gene in endosperm, but also in the embryo, cotyledons and pollen. The first 185 bp of the O2 promoter is sufficient for developmentally regulated expression in tobacco seeds. A distinct cis-acting element, located between positions -185 and -520, directs expression in the cotyledons of tobacco seedlings. The possible origins of this breakdown in promoter specificity in the heterologous host are discussed.