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IntroductionDrought detection, spanning from early stress to severe conditions, plays a crucial role in maintaining productivity, facilitating recovery, and preventing plant mortality. While handheld thermal cameras have been widely employed to track changes in leaf water content and stomatal conductance, research on thermal image classification remains limited due mainly to low resolution and blurry images produced by handheld cameras. MethodsIn this study, we introduce a computer vision pipeline to enhance the significance of leaf-level thermal images across 27 distinct cotton genotypes cultivated in a greenhouse under progressive drought conditions. Our approach involved employing a customized software pipeline to process raw thermal images, generating leaf masks, and extracting a range of statistically relevant thermal features (e.g., min and max temperature, median value, quartiles, etc.). These features were then utilized to develop machine learning algorithms capable of assessing leaf hydration status and distinguishing between well-watered (WW) and dry-down (DD) conditions. ResultsTwo different classifiers were trained to predict the plant treatment—random forest and multilayer perceptron neural networks—finding 75% and 78% accuracy in the treatment prediction, respectively. Furthermore, we evaluated the predicted versus true labels based on classic physiological indicators of drought in plants, including volumetric soil water content, leaf water potential, and chlorophyllafluorescence, to provide more insights and possible explanations about the classification outputs. DiscussionInterestingly, mislabeled leaves mostly exhibited notable responses in fluorescence, water uptake from the soil, and/or leaf hydration status. Our findings emphasize the potential of AI-assisted thermal image analysis in enhancing the informative value of common heterogeneous datasets for drought detection. This application suggests widening the experimental settings to be used with deep learning models, designing future investigations into the genotypic variation in plant drought response and potential optimization of water management in agricultural settings.