Soybean is a widely planted transgenic crop worldwide. However, it is still one of the most recalcitrant crops for genetic transformation due to the genotype dependence and the infection ability of Agrobacterium. In this study, the GUS transient expression analysis was used for evaluating the Agrobacterium sensitivity for different soybean genotypes and four Agrobacterium strains. The highest transient expression efficiency of two soybean genotypes were acquired when the explants were infected by Ag10 among different stains. Meanwhile, Agrobacterium sensitivity of 33 soybean genotypes including modern cultivars, landrace and wild soybeans was assessed and six sensitivity genotypes were identified when compared with the sensitivity control Peking. The results also showed that the Agrobacterium sensitivity for landrace or wild soybean was significant higher than modern cultivars. These results are important to construct or improve the high-efficiency soybean genetic transformation system and to develop new receptors for high efficiency genetic transformation.