Abstract: MYB transcription factors perform significant regulatory functions in plant growth and development. In this research, using the transcriptome data obtained from the disease-resistant soybean germplasm Dongnong L-10 and the disease-sensitive germplasm Heinong 37, the differentially expressed gene GmMYB48 was screened, which was a member of the MYB family, by using the physiological microspecies of soybean cyst nematode race 3（SCN 3）. The GmMYB48 gene was cloned and subjected to bioinformatics, subcellular localization, gene expression pattern analysis and gene overexpression analysis. The results showed that the protein encoded by GmMYB48transcription factor was a basic protein, which was covalently related to several species and widely distributed in plants; the target gene prediction results showed that the GmMYB48 gene was involved in the response to salt stress, drought stress, metal salt ion stress and other adversity stresses in soybean; the promoter element analysis showed that there were several responses to the gene in response to the environmental stress response related to the WUN-motif, MYB, etc. The gene expression pattern was analyzed and the gene overexpression was performed. The transient expression in tobacco leaves showed that GmMYB48 was subcellularly localized in the nucleus;GmMYB48 responded to soybean cyst nematode, and the relative expression of the gene was higher in the roots of disease-resistant materials than that of susceptible materials, and the overexpressed soybean hairy roots were obtained through Agrobacterium root-mediated genetic transformation. The number of nematodes per unit area in the hairy roots was significantly reduced, and the involvement of GmMYB48 in the resistance response of soybean to cyst nematode race 3 was verified from the phenotypic characterization. In this study, it initially investigated the function of GmMYB48 in resistance to soybean cyst nematode, and laid the foundation for further researches on the mechanism of soybean cyst nematode resistance.