Cdc42, a member of the Rho-family small GTP-binding proteins, is a pivotal signaling switch that cycles between active GTP-bound and inactive GDP-bound forms, controlling actin cytoskeleton organization and cell polarity. In this report, we show that MgCdc42, a Cdc42 ortholog in Magnaporthe grisea, is required for its plant penetration. Consequently, the deletion mutants show dramatically decreased virulence to rice due to the arrest of penetration and infectious growth, which may be attributed to the defect of turgor and superoxide generation during the appressorial development in Mgcdc42 deletion mutants. In addition, the mutants also exhibit pleotropic defects including gherkin-shaped conidia, delayed germination as well as decreased sporulation. Furthermore, dominant negative mutation leads to a similar phenotype to that of the deletion mutants, lending further support to the conclusion that MgCdc42 is required for the penetration and virulence of M. grisea.