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Lake water quality is affected by local and regional drivers, including lake physical characteristics, hydrology, landscape position, land cover, land use, geology, and climate. Here, we demonstrate the utility of hypothesis testing within the landscape limnology framework using a random forest algorithm on a national‐scale, spatially explicit data set, the United States Environmental Protection Agency's 2007 National Lakes Assessment. For 1026 lakes, we tested the relative importance of water quality drivers across spatial scales, the importance of hydrologic connectivity in mediating water quality drivers, and how the importance of both spatial scale and connectivity differ across response variables for five important in‐lake water quality metrics (total phosphorus, total nitrogen, dissolved organic carbon, turbidity, and conductivity). By modeling the effect of water quality predictors at different spatial scales, we found that …