Abstract: In this study, the genetic diversity of 50 soybean germplasms from Huanghuaihai region was analyzed by using 42 pairs of SSR markers.The results showed that 203 bands were amplified from the genome of the tested materials, and there were 198 polymorphic bands and the percentage of polymorphic loci（PPB） was 97.54%.The average number of alleles（Na）,the average number of effective alleles（Ne） and the average Nei’s gene diversity index（H） were 1.952 4,1.417 2 and 0.267 2 respectively.Additionally, the genetic diversity（Ht）,intra-population genetic diversity（Hs）,inter-population genetic differentiation coefficient（Gst） and gene flow（Nm*） were 0.270 4,0.259 1,0.141 7 and 3.03,respectively.The Na, H and Shannon’s information index（I） of soybean germplasms in the northern of Huanghuaihai region were higher than those in the southern part of Huanghuaihai region.The results showed that 42 pairs of SSR primers had abundant polymorphic information.UPMGA clustering analysis showed that 50 soybean germplasms from Huanghuaihai were divided into two groups, which were basically consistent with the structure of the two populations.The genetic distance between populations was small, and the similarity coefficient was large, indicating the medium and low degree of genetic differentiation.There was abundant gene exchange among populations, indicating there being abundant gene exchange between populations.The genetic diversity of soybean germplasm resources in the northern part of Huanghuaihai was higher than that in the southern part of Huanghuaihai, and the soybean germplasms in the northern part of Huanghuaihai were richer.