This quantitative study analyzed and predicted gender differences of agriculture graduates' attitudes towards and challenges in entrepreneurship in Botswana. The study adopted a descriptive and correlational survey research design. A valid and reliable questionnaire was used for data collection through a survey of randomly sampled 149 final year agriculture graduate students (n=149). Inferential statistical tools of Independent t-test and Regression analysis were used for data analysis. The findings of the study determined three important attitudinal factors as: entrepreneurship results in economic growth of a country, employability and income generation and, entrepreneurship improves individual and social growth. Three important constraints in entrepreneurship were lack of land, lack of proper infrastructure and, lack of capital. These top attitudinal factors as well as constraints were the same for the male and female graduates despite of their ranking and importance. A gender difference in students' attitudes towards entrepreneurship was established while no gender difference in the challenges in entrepreneurship was found. Out of fifteen constraints in entrepreneurship under study, only three constraints namely, lack of land, high competition in market and lack of capital, were determined as significant predictors of the graduates' attitudes towards entrepreneurship. It is recommended that these three factors be made priorities while making policies for entrepreneurship development in the country. Further study is recommended to explore the perceptions of graduates on the possible ways to improve on these three predicting constraints and explore latent constraints predicting graduates' attitude towards entrepreneurship. Those findings may provide better ideas in planning policies for entrepreneurial development among agriculture graduates in Botswana.