T5/ T6 generation of JNhrpZPsta2004-30-384 transgenic plants were used as the source of plant materials. The molecular detection and stability identification of T5 and T6 soybean lines were carried out by PCR, Southern blot and qRT-PCR techniques, and then indoor artificial inoculation was used for disease resistance analysis. The results show that the CaMV35S promoter, Nos polyA terminator, target gene and Bar gene were expressed in transformed soybean, indicating that the target gene was stably inherited. The Southern blot assay revealed that multiplecopy of the fragment was inserted in the transformed soybean, and the site was different. The qRT-PCR detection results indicate that the target gene was expressed in all organs (leaves, shoots and roots) of the transgenic soybeans, with the highest expression level in leaves and the lowest in roots. The hrpZPsta expression of roots, shoots and leaves in T5 generation was 2.871, 1.363 and 7.743， respectively, and the hrpZPsta expression of roots, shoots and leaves in T6 generation was 3.577, 1.664 and 8.589, respectively. In accordance with the “identification of soybean gray spot disease technical specifications”, the results suggest that the disease index of T5/T6 untransformed soybean was 52.4% and 50.6%, respectively, and the disease index of T5/T6 transformed soybean was 38.7% and 38.4%, respectively. The untransformed soybean belonged to the medium susceptibility level and transgenic soybean belonged to the medium resistance level.