Protein phosphatase 2Cs (PP2Cs) have been demonstrated to play critical roles in regulation of plant growth/development, abscisic acid signaling pathway and adaptation to environmental stresses. Here we report the cloning and molecular characterization of a novel rice protein phosphatase 2C gene, OsBIPP2C2 (Oryza sativa L. BTH-induced protein phosphatase 2C 2). OsBIPP2C2 has three alternatively spliced transcripts and the largest transcript OsBIPP2C2a encodes a 380 aa protein containing all 11 conserved catalytic subdomains of PP2Cs. Expression of OsBIPP2C2a was significantly induced by benzothiadiazole (BTH), one of defense-related signal molecules in plants. Expression of OsBIP2C2a was induced by infection with the blast fungus, Magnaporthe grisea, and the pathogen-induced expression of OsBIPP2C2a in BTH-treated rice seedlings was much earlier and stronger than those in water-treated seedlings. Overexpression of OsBIPP2C2a in transgenic tobacco plants resulted in increased disease resistance against tobacco mosaic virus and Phytophthora parasitica var. nicotianae. Importantly, the OsBIPP2C2a-overexpressing transgenic tobacco plants showed constitutive expression of defense-related genes. These results suggest that OsBIPP2C2a may play an important role in disease resistance through activation of defense response.