Abstract: In order to detect the integration and genetic stability of NPR1 gene in soybean Jilin 30 transgenic CHR3 line, to investigate the disease resistance of transgenic progeny with bivalent gene of CHR3 and NPR1, and to provide an effective reference for breeding new soybean varieties resistant to Phytophthora root rot. In this paper, soybean varieties JL30+GmCHR3 were used as receptor materials, the broad-spectrum resistance gene NPR1 was introduced into the receptor by agrobacteria-mediated. Conventional PCR were used to test the genetic transformation, southern blot hybridization and qRT-PCR were used to identify the integration and expression of the two genes in the T₁ and T₂ generations of JL30 + GmCHR3 receptor and double-resistant transgenic lines repectively, and hypocotyl infection was used to identify the resistance of transgenic soybean plants to Phytophthora root rot. The results showed that promoter 35 s, terminator Nos, screening marker gene Bar and target gene NPR1 were all expressed in receptor genome. Southern blotting showed that the target gene was integrated in soybean plants in a single copy manner. And the qRT-PCR detection results showed that the target gene was expressed in the root, stem and leaf of soybean plant, among them, the average relative expression of roots, stems and leaves of T₁ generation were 2.732, 1.614 and 3.316, respectively, the mean relative expression of roots, stems and leaves of T₂ generation strains were 2.936, 2.084 and 3.864, respectively. The results showed that the relative expression of the target gene in each tissue was as stem ＜ root ＜ leaf. Transformation lines with dual resistance genes of CHR3 and NPR1 showed high resistance to Phytophthora root rot disease, JL30+NPR1 showed moderate resistance, and non-transgenic plants showed susceptible. The results showed that the transformation of the bivalent resistance genes GmCHR3 and NPR1 into the soybean Jilin 30 could enhance the resistance to Phytophthora root rot.