3 strains of Nomuraea rileyi from different regions(Shandong，Anhui，Henan)were cultured on PDA and their virulence to the 3rd Spodoptera litura larvae were compared. Results show that, the 3 strains from different regions have the similar colony morphology, while there was remarkable differences in sporulation. The highest sporulation of strains was Nr19 from Anhui, the production of spore was 1.64×108 spores/cm2; In the toxicity tests, three of them had obvious pathogenic effects on the S. litura larvae, the mortality and cadaver rates reached more than 95% and 80%, respectively. And the Nr19 had the best comprehensive pathogenic effects on the 3rd Spodoptera litura larvaes, the corrected mortality and muscardine cadaver were 97.62% and 84.09%, respectively; the median lethal time (LT50) was 4.0441 d.