In order to understand the relationship between the distribution of epilithic diatoms and the habitual environments, land-use, water qualities, and epilithic diatoms were studied at 141 sampling sites in the midwestern stream of Korean peninsula (Geum river, Mangyeong river, Dongjin river, and Sapgyo river). The total 183 diatom taxa was appeared in the study, while the dominant species were found to be Nitzschia palea (10.9%) and Achnanthes convergens (8.4%). Based on the abundance of epilithic diatoms, a cluster analysis results indicate that the sampling sites divided the sampling sites into 4 groups (G) at the 25% level. In term of geographic and aquatic environments, G-I and -II accounted for the upper and mid streams of the Geum river, and had large forest areas and good in water quality. G-III accounted for farmland and urban, and high concentration nutrient levels (TN and TP) and electric conductivity. G-IV accounted for mostly farmland, and high levels in turbidity, BOD, nutrient and electric conductivity. CCA results showed that the saproxenous taxa Meridion circulare was the indicator species of G-I, which strongly influenced by altitude and forests. In G-II, the indifferent taxa Navicula cryptocephala was influenced by Chl-a, AFDM, and DO. In G-III and -IV, the indifferent taxa Fragilaria elliptica and saprophilous taxa Aulacoseira ambigua were influenced by electric conductivity, turbidity, and nutrient counts. Meanwhile, random forest results showed that the predicting factor of indicator species appearance in G-I, -II, and -III was found to be electric conductivity whereas in G-IV it was found to be turbidity. Collectively, the distribution of diatoms in the midwestern of Korean peninsula was found to depend more on the land-use and its subsequent water qualities than the inherent characteristics of the aquatic environment.