BACKGROUND: Education by smartphone-based software is a new method among the world's educational practices. Due to the lack of studies on the effect of this education on promoting knowledge and self-care behaviors of students’ reproductive health this study aimed to determine the effect of smartphone-based software education on knowledge and self-care behaviors in the field of reproductive health. MATERIALS AND METHODS: This RCT with 164 married female students (82 in each group) conducted from October 2019 to January 2020, in Tabriz University and Tabriz medical University. Participants were randomly allocated to two groups of educational intervention and control. Smartphone-based software education designed in four areas of reproductive health. Intervention group was trained for 4 weeks. The score of knowledge and self-care behaviors of participants in both groups were compared at baseline and 8 weeks after intervention. Data were analyzed by SPSS-23 software using Chi-square, independent t-tests, and analysis of covariance tests. RESULTS: Eight weeks after the intervention, a significant increase was observed in the mean (standard deviation [SD]) score of overall knowledge for reproductive health and all its subdomains (P < 0.001), as well as in in the mean (SD) self-care score of reproductive health and all subdomains (P < 0.05), except for the family planning in the education group compared to the control group. CONCLUSION: The findings indicate a significant effect of smartphone-based software on increasing students’ knowledge and self-care in all areas of reproductive health except for family planning subdomain of self-care.