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Rivers are under increasing pressure from anthropogenic impacts with incremental dam construction, experiencing global and regional alteration due to river disconnectivity, flow regulation, and sediment reduction. Assessing the cumulative impacts of dams on river disconnectivity in large river basins can help us better understand how humans disintegrate river systems and change the natural flow regimes. Using the Yangtze basin as the study area, this study employed three modified metrics (river connectivity index, RCI; basin disconnectivity index, BDI; and the degree of regulation for each river section, DOR) to evaluate the cumulative impacts on river disconnectivity over the past 50 years. The results indicated that the Yangtze had experienced strong alterations, despite varying degrees and spatial patterns. Among the major tributaries, the greatest impact (lowest RCI value) happened in the Wu tributary basin due to the construction of cascade dams on the main stem of the tributary, while the lowest impact (highest RCI value) happened in the Fu tributary basin, which still has no dams on its main stem. Collectively, rivers in the upper Yangtze reaches experienced more serious disturbances than their counterparts in the middle and lower reaches. The BDI results displayed that a substantial part of the Yangtze River, especially the Wu, Min, Jialing, and Yuan tributaries, only maintain connectivity among one to three representative river systems. No part of the Yangtze connects all the 12 representative river systems. This study also revealed that small dams can also exert significant impacts in flow regulation on regional river systems through their sheer number and density. The study results can help promote more environmentally sustainable river management policies in the Yangtze basin.