We present a continent-scale exploration of trends in annual 7-day low streamflows at 2482 U.S. Geological Survey streamgages across the conterminous United States over the past 100, 75, and 50 years (1916–2015, 1941–2015 and 1966–2015). We used basin characteristics to identify subsets of study basins representative of reference basins with streamflow relatively free from human effects (n = 259), and predominantly agricultural basins (n = 78), regulated basins (n = 220), and urban basins (n = 121). Trend significance was computed using the Mann-Kendall test considering short- and long-term persistence. Lag-one autocorrelation tests of detrended 7-day low streamflows for all gage classes show that time-series independence is not an appropriate assumption for annual low streamflow data at many basins. Among all study gages, upward trends (wetter conditions) in 7-day low streamflows outnumbered downward trends (drier conditions) approximately 2–1 for the 75- and 100-year trend periods—50-year trends indicated roughly equal numbers of increases and decreases. Increases in 7-day low streamflow were consistently observed for all time periods throughout much of the northeastern quadrant of the conterminous U.S. including western New England and the Mid-Atlantic, the southeastern Great Lakes basin, northern Ohio River basin, and the Upper Mississippi River and eastern Missouri River basins. Decreases in 7-day low streamflow were consistently observed for all time periods at many gages in the southeastern U.S. and in the northwestern U.S. in much of Idaho and northwestern Washington. Overall, we observed greater percentages of statistically significant trends at gages with human-induced influences than at reference gages. Low-flow trends at agricultural gages were regionally consistent with trends at reference gages. Regulated basins had many statistically significant upward trends for all three time periods tested, which may be attributed in part to substantial increases in dam-related storage prior to 1970. Urban gages had the greatest percentage of significant decreases in 7-day low flows compared to all other gage classes even though most urban gages saw upward trends in mean annual flows. Urban gages also had the greatest percentage of significant increases in low flows second only to regulated gages, highlighting that urban development can increase or decrease low streamflows depending on the basin-specific development.