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This study investigated the main sources of Iranian students' demotivation in L2 learning and examined demotivation in relation to students' gender and level of education. To find the major demotivating factors, a questionnaire consisting of 32 items was developed and completed by 327 students. An exploratory factor analysis was conducted to explore the factorial structure of the questionnaire. Based on the results, five categories of demotivating factors were identified: Learning Contents, Materials, and Facilities, Attitude towards English Speaking Community, The Teacher, Experience of Failure, and Attitude towards Second Language Learning. To examine demotivating factors in relation to students' gender and educational level, two one-way analyses of variance were run. The results indicated significant differences between male and female students in terms of three demotivating factors. Significant differences were observed between students at different levels of education with regard to the three demotivating factors. Finally, a multivariate analysis of variance was performed to examine demotivating factors in relation to students' gender and educational level. Significant differences were found between male and female students across educational levels in terms of all demotivating factors except one.