The low prevalence of Exclusive Breastfeeding in Yogyakarta City is found in the working area of Public Health Center Pakualaman that is 10,9% and in Gunungkidul Regency, it is found in the working area of Public Health Center Wonosari 2 that is 27,6% (DINKES DIY, 2014). The related data is still low if it is compared to the prevalence goal target which is set by The Ministry of Health that the cover of the Exclusive Breastfeeding to 0-6 month babies should be 80%. The high knowledge of the mothers on Exclusive Breastfeeding will motivate the mothers to give Exclusive Breastfeeding to their babies. Due to the low number of Exclusive Breastfeeding, the differences between the mother’s knowledge on Exclusive in Urban and Rural are needed to find. This research aims to find out the differences between the mother’s knowledge on Exclusive Breastfeeding in Urban and Rural. Research Method: this research is non-experimental using descriptive comparative research design. The research samples are 45 mothers living in the working area of Public Health Center Pakualaman and 83 mothers living in the working area of Public Health Center Wonosari 2. The sampling technique is a random sampling. The research data analysis uses univariate analysis thet is the distribution frequency and bivariate analysis that is Mann-Whitney test. Research Result: In cities, there are 42 respondents (93,3%) who have high knowledge level on exclusive breastfeeding and 3 respondents (6,7%) who have moderate knowledge level on Exclusive Breastfeeding. Meanwhile in Rural, there are 71 respondents (85,5%) who have high knowledge level on Exclusive Breastfeeding and 12 respondents (14,5%) who have moderate knowledge level on Exclusive Breastfeeding. Mann-Whitney test shows the result of p=0,306 (p>0,05). Conclusion: there is no difference between the mother’s knowledge on Exclusive Breastfeeding in Urban and Rural