Using the Regulative Theory of Temperament, this study aimed to identify relationships between two temperamental traits – activity and emotional reactivity – and functional connectivity of the amygdala and the anterior cingulate cortex during the Affective Stroop task. Participants were 84 young adults (42 women) aged 19–25. Emotional reactivity was negatively related to connectivity between the right amygdala and the left lateral occipital cortex, right cerebellum, and left inferior frontal gyrus (IFG) as well as right rostral anterior cingulate cortex (rACC)–frontal pole connectivity. Temperamental activity was related to right amygdala–left subcallosal cortex positive connectivity; and right amygdala–left IFG, and left middle temporal gyrus (MTG) negative connectivity and rACC–cerebellum negative connectivity. The interaction between the two traits was related to the connectivity between the amygdala and right cerebellum, left subcallosal cortex, and left MTG, as well as right rACC–right prefrontal pole connectivity. Our results are in line with previous literature, suggesting diminished cognitive resources among individuals with ineffective regulatory temperamental systems. •Reactivity is related to connectivity between amygdala and lateral and frontal areas•Activity is related to negative coupling of rACC with cerebellum•Trait interaction is related to negative amygdala-cerebellum coupling•Lessen cognitive resources is a possible effect of ineffective temperamental regulation