Since the 1980's, it has been common practice for children to attend classes in their bare feet in Japan. However, no academic evidence in support of this practice exists. The aim of present study was to investigate the relationship between barefoot education and physical fitness and/or motor ability. The participants in this study were 153 preschoolers aged four and five years old. All the participants completed seven distinct motor ability tests and their toe-grip strength was measured via TKK-3360. A two-way layout ANOVA with general linear model (GLM) was applied to each gender, distinguishing between age and educational method (barefoot/shoes). As to the effect of the educational method, there was no difference apart from in relation to toe-grip strength and the side jump for boys (barefoot＜shoes), and the potato-race for girls (barefoot＞shoe). All effect sizes were shown to be less than 0.1 in absolute value. Physical fitness was aggregated into the following two components via a factor analysis process:“Muscle strength & Coordination (MSC)” and “Grasp reflex (GR)”. A multiple regression analysis with dummy variables was applied as a factor score for dependent variables and gender, age and educational method for independent variables. In terms of their MSC factors, gender (boy＞girl) and age (4yr.＜5yr.) displayed significant variables, while educational method (barefoot＜shoes) and age (4yr.＜5yr.) displayed significant variables in terms of GR. The results of the present study therefore revealed that the children whoeducated by barefoot were not superior in physical fitness and motor ability.