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Background: Root system architecture (RSA) exhibits significant genetic variability and is closely associated with drought tolerance. However, the evaluation of drought-tolerant cotton cultivars based on RSA in the field conditions is still underexplored. Results: So, this study conducted a comprehensive analysis of drought tolerance based on physiological and morphological traits (i.e., aboveground and RSA, and yield) within a rain-out shelter, with two water treatments: well-watered (75 ± 5% soil relative water content) and drought stress (50 ± 5% soil relative water content). The results showed that principal component analysis identified six principal components, including highlighting the importance of root traits and canopy parameters in influencing drought tolerance. Moreover, the systematic cluster analysis was used to classify 80 cultivars into 5 categories, including drought-tolerant cultivars, relatively drought-tolerant cultivars, intermediate cultivars, relatively drought-sensitive cultivars, and drought-sensitive cultivars. Further validation of the drought tolerance index showed that the yield drought tolerance index and biomass drought tolerance index of the drought-tolerant cultivars were 8.97 and 5.05 times higher than those of the drought-sensitive cultivars, respectively. Conclusions: The RSA of drought-tolerant cultivars was characterised by a significant increase in average length-all lateral roots, a significant decrease in average lateral root emergence angle and a moderate root/shoot ratio. In contrast, the drought-sensitive cultivars showed a significant decrease in average length-all lateral roots and a significant increase in both average lateral root emergence angle and root/shoot ratio. It is therefore more comprehensive and accurate to assess field crop drought tolerance by considering root performance. [ABSTRACT FROM AUTHOR]