Early childhood is a critical period for the development of fundamental movement skills (FMS). Children who do not master FMS are more likely to experience failure in the motor domain and less likely to participate in sport and games during childhood and adolescence. Studies among primary school aged children report low levels of FMS mastery indicating the need to implement FMS programs during the preschool years. Cross-sectional study of 425 children attending preschools in the Sydney, Australia in 2008. FMS were assessed using the Test of Gross Motor Development-2 including locomotor (run, gallop, hop, horizontal jump) and object control (strike, catch, kick overhand throw) skills. Data were analysed using linear regression and chi-squared analyses. Total locomotor score was higher among girls compared with boys (p<0.00); however only the hop was significantly different (p=0.01). Boys had higher total (p<0.00) and individual object control scores compared with girls, except the catch (p=0.6). The prevalence of mastery differed across each FMS. Girls generally had higher mastery of locomotor skills and boys had higher mastery of object control skills. These findings highlight the need to provide structured opportunities which facilitate children's acquisition of FMS, which may include providing gender separated games, equipment and spaces. That mastery of FMS is low in primary school children indicates the importance of early intervention programs in preschools. Preschools and child care centers hold promise as a key setting for implementing FMS programs.