The objective of this study was to assess the relationship between imaging surrogates for diffuse fibrosis and myocardial dysfunction. Thirty-six New Zealand white rabbits were classified into two groups: a control group (n = 18) and an alloxan-induced diabetes mellitus (DM) group (n = 18). For all rabbits, conventional ultrasonography, two-dimensional speckle tracking, and cardiac magnetic resonance (CMR) T1 mapping were performed; all of the rabbits were then sacrificed for Masson's staining. The extracellular volume (ECV) was calculated from pre-and post-contrast T1 values and compared with myocardial function measured by echocardiography using Pearson's correlation. In the DM group, ECV increased as the duration of diabetes increased, consistent with the changes in myocardial fibrosis verified by pathology. Moreover, ECV was strongly correlated with the early diastolic strain rate (r = -0.782, p < 0.001) and moderately correlated with the radial systolic peak strain (r = 0.478, p = 0.045). Thus, ECV is an effective surrogate for myocardial diffuse fibrosis on CMR imaging, and higher ECV values are associated with an increased impairment of myocardial diastolic function.