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Uniconazole (UNZ) can alleviate a variety of abiotic stresses such as low temperature. With application of UNZ on Coix lachryma-jobi L. (coix) under low-temperature stress, growth and physiological parameters were investigated in seedlings. Meanwhile, transcriptome profile in coix seedlings was characterized as well. The results showed an increase of 11.90%, 13.59%, and 10.98% in stem diameter, the aboveground and belowground biomass in 5 mg/L uniconazole application group (U3), compared with control check low-temperature group (CKL). Some anti-oxidase activities also show significant difference between CKL and U3 ( p  < .05). Transcriptome results showed that 3,901 and 1,040 genes had different expression level at control check (CK) and CKL, CKL and U3. A considerable number of different expressing genes (DEGs) related to the plant hormone signal transduction, photosynthesis, reactive oxygen species (ROS)-related genes, and secondary metabolism in response to uniconazole application were identified in this study. The transcriptomic gene expression profiles present a valuable genomic tool to improve studying the molecular mechanisms underlying low-temperature tolerance in coix. At the same time, it would provide a certain basis for the application of UNZ in the production of coix resistance under low temperature., Competing Interests: The authors declare that they do not have any conflict of interest., (© 2019 The Authors. Food Science & Nutrition published by Wiley Periodicals, Inc.)