Although previous studies have documented a variety of electrocardiogram abnormalities in beta-thalassemia major (β-TM), little is known about P-wave dispersion (PD), an independent risk factor for development of atrial fibrillation. The aim of our study was to evaluate PD in β-TM patients with conserved systolic and diastolic functions. The study involved 40 β-TM patients (age 37.5 ± 10.2; 33 M) and 40 healthy subjects used as controls, matched for age and gender. PD was carefully measured using a 12-lead electrocardiogram. Cardiac iron levels were measured by cardiac magnetic resonance T2 star (CMR T2*) imaging. Comparing to the healthy control group, β-TM group presented increased values of the PD (40.1 ± 12.9 vs. 24 ± 7 ms; P < 0.004) and decreased CMR T2* imaging (29 ± 15 vs. 55 ± 13 ms; P = 0.03). We found a significant correlation between PD and CMR T2* values. Our study showed a significant increase of PD in β-TM patients with conserved systolic and diastolic cardiac functions. Our results indicate that PD is correlated to myocardial iron deposit, as assessed by CMR T2* imaging.