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This study reports the preliminary results from a statistical screening of tree-ring width records from the International Tree-Ring Data Bank (ITRDB), to evaluate the strength of the hydrological signal, in dendrochronological records from the Tennessee Valley. We used United States Geological Survey (USGS) streamflow data from 11 gages, within the Tennessee Valley, and regional tree-ring chronologies, to analyze the dendroclimatic potential of the region, and create seasonal flow reconstructions. Prescreening methods included correlation, date, and temporal stability analysis of predictors to ensure practical and reliable reconstructions. Seasonal correlation analysis revealed that large numbers of regional tree-ring chronologies were significantly correlated (p ≤ 0.05) with the May–June–July streamflow. Stepwise linear regression was used to create the May–June–July streamflow reconstructions. Ten of the 12 streamflow stations were considered statistically skillful (R2 ≥ 0.40). Skillful reconstructions ranged from 208 to 301 years in length, and were statistically validated using leave-one-out cross validation, the sign test, and a comparison of the distribution of low flow years. The long-term streamflow variability was analyzed for the Nolichucky, Nantahala, Emory, and South Fork (SF) Holston stations. The reconstructions revealed that while most of the Western United States (U.S.). was experiencing some of its highest flow years during the early 1900s, the Tennessee Valley region was experiencing a very low flow. Results revealed the potential benefit of using tree-ring chronologies to reconstruct hydrological variables in the Southeastern U.S., by demonstrating the ability of proxy-based reconstructions to provide useful data beyond the instrumental record.