The functions of four loci (An1, An2, An4, and An6) which control pigmentation in flowers of Petunia hybrida have been characterized. Linkage-analysis and molecular complementation experiments showed that the An6 locus contains the structural dfrA gene, encoding the enzyme dihydroflavonol 4-reductase (DFR). Analysis of gus gene expression driven by the dfrA promoter in transgenic plants showed that the dfrA promoter is highly active in the flower corolla, the anthers and seeds and, at a lower level, in ovules and the flower stem. These data are discussed in relation to the expression of other pigmentation genes and the accumulation pattern of anthocyanins. The expression of the drfA-gus transgene was dependent on the genes an1 (in every tissue), an2 (in the flower limb only) and an4(in anthers), demonstrating that these genes encode regulatory factors that control drfA promoter activity.