An optimal level of dopamine (DA) in the mammalian prefrontal cortex (PFC) is critical for higher cognitive control of behavior. Too much or too little DA in the PFC induces impairment in working memory (WM) task performance. PFC DA is also concerned with motivation. When reward is anticipated and/or delivered, an increase in PFC DA release is observed. In the primate, more preferred reward induces enhanced WM-related neuronal activity in the dorsolateral PFC (DLPFC). We hypothesized that there would be more DA release in the primate DLPFC when more preferred, as compared with less preferred, reward is delivered during a WM task. Contrary to our hypothesis, we found higher DA release in the DLPFC when less rather than more preferred reward was used during a WM task, while unpredictable free reward delivery induced an increase in DLPFC DA release irrespective of the difference in the incentive value of the reward. Behaviorally, the monkey was more motivated with preferred than with less preferred reward, although it performed the task almost without error irrespective of the difference in the reward. Considering that mild stress induces an increase in DA release in the mammalian PFC, performing a WM task for less preferred reward could have been mildly stressful, and this mild stress may have induced more DLPFC DA release in the present study. The higher DA release in the DLPFC with less preferred reward may be beneficial for monkeys to cope with mildly stressful and unfavorable situations to achieve proficient WM task performance.