Working memory (WM) capacity is associated with various emotional aspects, including states of depression and stress, reactions to emotional stimuli, and regulatory behaviors. We have previously investigated the effects of WM training (WMT) on cognitive functions and brain structures. However, the effects of WMT on emotional states and related neural mechanisms among healthy young adults remain unknown. In the present study, we investigated these effects in young adults who underwent WMT or received no intervention for 4 weeks. Before and after the intervention, subjects completed self-report questionnaires related to their emotional states and underwent scanning sessions in which brain activities related to negative emotions were measured. Compared with controls, subjects who underwent WMT showed reduced anger, fatigue, and depression. Furthermore, WMT reduced activity in the left posterior insula during tasks evoking negative emotion, which was related to anger. It also reduced activity in the left frontoparietal area. These findings show that WMT can reduce negative mood and provide new insight into the clinical applications of WMT, at least among subjects with preclinical-level conditions.