Dihydroflavonol-4-reductase (DFR) catalyzes the reduction of dihydroflavonols in the anthocyanin biosynthetic pathway. In this study, we obtained the full-length cDNA of DFR from Calibrachoa hybrida, CaDFR, through RACE-based technique. The cDNA of CaDFR is 1,351 bp with an open reading frame of 1, 152 bp, which encodes a putative protein of 383 amino acids. The CaDFR protein is predicted with 42.74 KD in molecular weight and a pI value of 5.83. Sequence alignment showed that CaDFR had close relationship with DFRs from other species of the Solanaceae family, and it belonged to the Asp-type. We constructed 35S::CaDFR overexpression vector and transformed it into three cultivars of Petunia hybrida using the Agrobacterium-mediated method. Multiple transformants were selected in the presence of 50 mg/l kanamycin antibiotic and further verified by polymerase chain reaction and Southern blotting analyses. We observed that the bud and flower color of transgenic petunia altered comparing with that of control plants. The DFR activities varied in most color-changed transgenic petunias. These results confirm the identity and function of CaDFR and further suggest that integration of the CaDFR can be used to change flower color of transgenic plants. (C) 2013 Elsevier B.V. All rights reserved.