Soil seed bank, generally acknowledged as an important seed reserve for vegetation regeneration, plays an important role in the vegetation restoration and succession, as well as the ecosystems construction. The species composition, spatio-temporal distribution, and quantitative changes of re-vegetated desert areas in Hexi Corridor were investigated in this study. Results showed that there were 27 species of plants in the soil seed bank, belonging to 22 genera and 8 families, with the most common species in Chenopodiaceae. In terms of life-form, the seeds from herbs accounted for 90.6-95.06%. The density of soil seed bank was between 19.29-858.57 grains/m², showing a horizontal distribution pattern from east to west. The soil seed banks was mainly concentrated in the 0-2 cm soil layer in vertical direction. In the rang of 0-10 cm soil layer, there was an obvious vertical distribution pattern of seed density:with increase of soil depth, seed density first decreased then increased. Simpson Index in the soil seed bank was the highest in the re-vegetated areas of the east Hexi Corridor Desert (0.671-0.812), 0.417-0.809 of the middle Hexi Corridor Desert, and 0.256-0.707 of west Hexi Corridor Desert, with a downward trend from east to west. Shannon-Wiener Index, Margalett Richness Index and Peilow Evenness Index also showed the same trend. The large variation in species diversity indices among sites was most likely attributed by the high degree of inter-site vegetation heterogeneity. The similarity coefficients between soil seed bank and vegetation in the desert re-vegetated areas in Hexi Corridor were all below 0.45, indicating a significant difference in species composition. The soil seed bank was dominated by annual and perennial herbs, with a few shrubs, which indicated that the soil seed bank played a minor role in the regeneration and succession of the shrub layer in the desert re-vegetated ecosystems. This study proved that a sufficient amount of seeds in soil seed bank is an important foundation for the natural restoration of vegetation in desert, and advanced our understanding of the natural vegetation restoration in the re-vegetated desert areas.