Background: Plants, involved in highly complex and well-coordinated system, have evolved a considerable degree of developmental plasticity, thus minimize the damages caused by stresses. MicroRNAs (miRNAs) have recently emerged as key regulators in gene regulation, developmental processes and stress tolerance in plants. Results: In this study, systematic discovery of soybean miRNAs associated with stress response (mock, drought, salinity and alkalinity) has been identified and analyzed in combination with deep sequencing technology and in-depth bioinformatics analysis. We found that a total of 133 conserved miRNAs corresponding to 95 miRNA families have expressed in soybean under four diverse treatments. In addition, 71, 50 and 45 miRNAs are either uniquely or differently expressed under drought, salinity and alkalinity respectively, suggesting that many miRNAs are inducible and are differentially expressed in response to certain stress. Conclusion: Our study has important implications for further identification of gene regulation under abiotic stresses and also significantly contributes a complete profile of miRNAs in Glycine max.