We identified the sites for the initiation of transcription of a gene for subunit 1 of F1-ATPase (atp1) in rice mitochondrial DNA. Capping and ribonuclease protection experiments in vitro, together with primer extension analysis, demonstrated that there were at least eight transcription initiation sites upstream of atp1. One initiation site, expressed most actively, was flanked by a sequence identical to the consensus promotor motif of rice mitochondrial genes. However, the sequences surrounding the other seven initiation sites exhibited no similarity to the consensus sequence.