Formic acid disrupts mitochondrial electron transport and sequentially causes cell death in mammalian ocular cells by an unidentified molecular mechanism. Here, we show that a low concentration of formic acid induces apoptosis-like cell death in the budding yeast Saccharomyces cerevisiae, with several morphological and biochemical changes that are typical of apoptosis, including chromatin condensation, DNA fragmentation, externalization of phosphatidylserine, reactive oxygen species (ROS) production, loss of mitochondrial membrane potential and mitochondrion destruction. This process may not be dependent on the activation of Yca1p, the yeast caspase counterpart. In addition, the cell death induced by formic acid is associated with ROS burst,while intracellular ROS accumulate more rapidly and to a higher level in the YCA1 disruptant than in the wild-type strain during the progression of cell death. Our data indicate that formic acid induces yeast apoptosis via an Yca1p-independent pathway and it could be used as an extrinsic inducer for identifying the regulators downstream of ROS production in yeast.