Background This study investigates the relationship between temperament traits postulated by Strelau’s regulative theory of temperament (RTT) and Akiskal’s affective temperaments. This study represents the first attempt to compare these two concepts in a non-clinical Polish sample. Participants and Procedure The study involved 615 healthy Caucasian adults (395 women and 220 men) aged from 17 to 69 years (M = 30.79, SD = 9.69). Temperament traits postulated by the RTT were assessed with the Formal Characteristics of Behaviour–Temperament Inventory. The Polish version of the Temperament Evaluation of Memphis, Pisa, Paris and San Diego Auto-questionnaire (TEMPS-A) was used to assess affective temperaments (depressive, cyclothymic, hyperthymic, irritable and anxious). Results Emotional reactivity and perseveration (RTT) positively correlated with anxious, cyclothymic, depressive and irritable temperaments (TEMPS-A), predicting from 2% (irritable temperament) to 24% (anxious temperaments) of the variance in the affective temperaments. Hyperthymic temperament (TEMPS-A) positively correlated with briskness, sensory sensitivity, endurance and activity (RTT). Activity was the best predictor of hyperthymic temperament, accounting for 25% of the variance. TEMPS-A scores showed that women were more depressive, cyclothymic and anxious and less hyperthymic than men. Conclusions These results suggest that two RTT traits, emotional reactivity and perseveration, may be related to all the affective temperaments of TEMPS-A, except hyperthymic temperament, which is most likely linked to the RTT activity trait.