In semiarid regions, deficit and unpredictable precipitation results in yield losses. Uniconazole is a plant growth regulator and its application is beneficial in water saving agriculture and improves maize production in semiarid regions. In order to determine the effects of uniconazole application on seed filling and hormonal changes of maize, a field study was conducted in the summer of 2015 and 2016. Seeds were soaked in uniconazole at concentration of 0 (SCK), 25 (S25), 50 (S50), and 75 (S75) mg kg-1, while in the second experiment, uniconazole was applied to the foliage at concentration of 0 (FCK), 25 (F25), 50 (F50), and 75 (F75) mg L-1 at the eight-leaf. Uniconazole application significantly improves the seed filling rates by regulating the endogenous hormones contents. Uniconazole seed soaking treatments improved significantly the seed filling rate of superior, middle, and inferior seeds compared with foliar application treatments. Uniconazole improved significantly the zeatin (Z) + zeatin riboside (ZR) and abscisic acid (ABA) contents while reducing the gibberellic acid (GA) content in the seeds during the process of seed filling. The Z + ZR and ABA contents were significantly positively correlated while the GA content was negatively correlated with maximum seed weight, maximum seed filling rates, and mean seed filling rates. Treatments S25 and F25 significantly improved the above dry matter accumulation plant-1, seed filling rates, ABA, Z + ZR contents, characters of ear, and grain yield while reduced the GA content. It is concluded from our results that the uniconazole application at concentration of 25 mg kg-1 as seed soaking or 25 mg L-1 foliar applied at the eight-leaf stage is beneficial to improve the seed filling rates and grain yield of maize in semiarid regions.