The molecular weight of the polysaccharide and the chemical groups it contains has an important influence on its biological activity, relatively low molecular weight polysaccharides may have better antioxidant activity. Polysaccharides isolated from the fruiting bodies of Morchella sextelata (MSP) were treated by γ-irradiation at 10, 100 and 1000 kGy doses, and the physicochemical properties and antioxidant activity of irradiated MSP were investigated. Microscopic observation under a scanning electron microscope (SEM) showed that breakage and pores appeared on the surface of the irradiated polysaccharide. As the irradiation dose increased, the average molecular weight of MSP decreased significantly, while the particle size and thermal stability of MSP first increased at 10 and 100 kGy doses and then decreased at 1000 kGy doses. The antioxidant activities, measured by free radical scavenging tests and protective effect on PC12 cells injured by H2O2, were all increased after irradiation, especially when the concentration of MSP was low (50 and 100 μg/ml). Therefore, irradiation treatment was an effective method to enhance the activity of polysaccharides.