Dabry′s Sturgeon (Acipenser dabryanus) is an endemic freshwater fish in the mainstream of upper reaches of the Yangtze River and its tributaries in western China. Due to anthropogenic factors, the natural populations of Dabry′s Sturgeon have been declining sharply in the past decades. Consequently, the species was listed as a first class protected animal by the Chinese government. And it was also listed as critical endangered in the IUCN Red List. In order to obtain the information on genetic diversity of Dabry′s Sturgeon, twenty-five tetranucleotide microsatellite loci were developed from the genomic DNA using the high-throughput sequencing. The polymorphism of microsatellites was tested using 18 wild individuals and 30 artificially propagated individuals. A total of 181 alleles were found in 25 microsatellites. The number of alleles per locus (A) ranged from 4 to 11 (mean = 7.2). And the observed heterozygosities (HO) and expected heterozygosities (HE) ranged from 0.160 to 1.000 (mean = 0.744) and 0.346 to 0.875 (mean = 0.727), respectively. No loci significantly deviated from the Hardy-Weinberg equilibrium (P > 0.05) and there was no significant linkage disequilibrium found between pairs of loci. All of the microsatellite markers, except one, showed polymorphism information content (PIC) values above 0.5 (Table 1). These polymorphic microsatellite loci described here will facilitate further studies on the population genetic management and conservation of Dabry′s sturgeon. In this article, Table 1 shows the information of polymorphic microsatellite markers isolated from Dabry′s Sturgeon.