This study explored gender differences on university students' attitudes toward Web-based learning in Taiwan. A Web-based Learning Attitude Survey (WLAS) was developed. The WLAS consisted of five scales, including access, social structure, content, pedagogy, and community relationship toward Web-based learning. By using an online survey, this study gathered the responses of 1,866 Taiwanese university students (940 males and 926 females) whose ages ranged from 18 to 23. The results showed significant gender differences in the aspects of social structure, content, and community relationship involved in Web-based learning. Interestingly, although females might consider the Internet a masculine-dominated technology, they displayed more favorable attitudes than males did regarding the impacts of socioeconomic status (the social structure scale) on students' performance of Web-based learning. Moreover, females possessed more positive attitudes than males toward the helpfulness and variety of the content for Web learning. However, they did not think the virtual interaction on the Internet could develop adequate teacher-student and peer relationships (the community relationship scale). Some follow-up interviews were also conducted with selected students, and the implications derived from this study were discussed.