• Premise of the study: A set of expressed sequence tag (EST) microsatellite markers were developed and characterized using next‐generation sequencing technology for the Chinese genus Ostryopsis (Betulaceae).• Methods and Results: A total of 38 high‐quality simple sequence repeat (SSR) primers were identified, of which 15 could be successfully amplified. Subsequently, we selected 80 individuals to represent the three species of the genus to evaluate the efficacy of these markers for examining genetic diversity of each species in the future. We found that the number of alleles per locus ranged from one to nine, with an average of 3.8. The expected heterozygosity and observed heterozygosity per locus varied from 0 to 0.829 and from 0 to 1, respectively, with their respective mean values as 0.483 and 0.416.• Conclusions: These EST‐SSR markers will be useful for evaluating the range‐wide genetic diversity of each species and examining genetic divergence and gene flow between the three species.