This study investigated the effects of dispositional and experimentally induced perspective-taking (PT) on physiological attunement between romantic partners during a conflict resolution task. Young adult couples (N = 103 dyads) rated their trait PT 1 week prior to participating in a conflict resolution session with their romantic partner. Immediately before the conflict task, participants were given one of the following three instructions: to take their partner’s perspective (PT condition), to approach the conflict mindfully (mindfulness condition), or to focus on their own perspective regarding the conflict (control condition). Participants provided four saliva samples over the course of the laboratory session, and the samples were assayed for alpha-amylase to measure autonomic nervous system activity. Multilevel modeling results revealed that couples in the PT condition displayed greater autonomic attunement over the course of the conflict session compared to those in the other conditions. In addition, female partners’ dispositional PT enhanced the effect of the PT induction on couples’ attunement. Furthermore, secondary analyses provided support for the beneficial role of autonomic attunement. Specifically, attunement was decreased by negative conflict behaviors and predicted increased post-conflict negative affect in females. Implications for dyadic functioning and intervention are discussed.