A wheat beta-1,3-glucanase gene (TaGluD) was identified as a fungal defense candidate. Its transcript induction was more than 60-fold higher in a resistant wheat line, Shannong0431, than in a susceptible wheat line, Wenmai6, after infection with Rhizoctonia cerealis. The TaGluD protein was overexpressed as inclusion bodies in Escherichia coli. After refolding and purification, TaGluD with 1 unit of beta-1,3-glucanase showed antifungal activity in vitro against Rhizoctonia solani, R. cerealis, Phytophthora capsici and Alternaria longipes with inhibition rates of 42%, 43%, 32% and 30%, respectively. Thus TaGluD may be useful for enhancing fungal resistance in several crop species.