Objective: To analyze the level of sex-related knowledge and behavior and related factors among children in senior grade of kindergarten in rural areas, and to provide the evidence for early childhood sexuality education. Methods: Multi-state stratified sampling method was adopted. From March to May 2019,a total of 324 students and their parents from 16 kindergartens of rural areas in Sichuan Province were enrolled, self-developed questionnaire was used to collect the basic demographic characteristics, information of sexuality education knowledge and sex-related behavior. Results: The average score of sexuality education knowledge was 5(4,6) point, and the awareness rate was 33.6%(109). The qualified sex-related behavior was observed in 75.0%(243 of all the children), while 25.0%(81) needed to be improved. Multivariate analysis showed that,compared with children were aware of the sexuality education knowledge, children who were unaware of the sexuality education knowledge were more likely to have good sex-related behavior, with the OR(95%CI) value was 2.06(1.14-3.72). The possibility of having good sex-related behavior among those children with more than 2 members in the family parents carring out sexuality education was higher than children without sexuality education in the family, with the OR(95%CI) value being 2.40(1.17-4.89). Conclusion The level of sexuality education knowledge among children in senior grade of kindergarten of rural areas in Sichuan is low, and most children s behavior needs to be improved. Children s awareness of sexuality education knowledge and the number of family members who have carried out sexuality education are related to the occurrence of children s sex-related behavior.