This study examined the potential relationship between participation in physical activity (PA) assessed by triaxial accelerometry and physical fitness testing, including health-related and skill-related parameters of fitness, in 136 Japanese preschoolers (65 girls and 71 boys, 5.5 ± 0.6 years). In partial correlation analyses, grip strength and 20m shuttle run test were positively correlated with time spent in physical activity ratio (PAR) ≥ 4. Better scores on standing long jump distance and jump over and crawl under tests were associated with lower sedentary time and greater moderate-to-vigorous PA time and PAR ≥ 4 time, and increased physical activity level. Moreover, 25m run speed was positively correlated with time spent in PAR ≥ 4 and locomotive activity. These findings suggest that development of both health-related (muscle strength and aerobic fitness) and skill-related fitness (power, agility and speed) may make engagement in PA easier for preschool children, although further research on the cause-effect relationship is needed.