Hypovitaminosis D has been associated with poorer physical and cognitive performances in older adults. The objectives of this study were (1) to measure and compare the time to perform (pTUG) and to imagine (iTUG) the Timed "Up & Go" test (TUG) test, and the time difference between these two performances (i.e., TUG delta time) in non-demented community-dwelling older adults with and without lower serum 25-hydroxyvitamin D (25OHD) concentrations and (2) to examine the association between the TUG delta time and serum 25OHD concentrations. Durations of pTUG, iTUG and TUG delta time, and serum 25OHD concentrations (severe insufficiency <10 ng/mL; moderate insufficiency: 10-30 ng/mL; normal status >30 ng/mL) were measured in 359 non-demented participants (mean age 70.4 ± 4.8 years; 40.7 % women). Participants with severe 25OHD insufficiency (15.6 %) had higher TUG delta time compared to those with moderate insufficiency (P = 0.010) and normal status (P = 0.048). TUG delta time was negatively associated with serum 25OHD concentrations (P < 0.010). Accurate motor imagery of gait was explained in part by serum 25OHD concentrations, increased discrepancy between pTUG and iTUG being associated with lower serum 25OHD concentrations.