<p dir="ltr">Recent advancements in hyperspectral imaging (HSI) for early disease detection have shown promising results, yet there is a lack of validated high-resolution (spatial and spectral) HSI data representing the responses of plants at different stages of leaf disease progression. To address these gaps, we used bacterial leaf spot (Xanthomonas perforans) of tomato as a model system. Hyperspectral images of tomato leaves, validated against in planta pathogen populations for seven consecutive days, were analyzed to reveal differences between infected and healthy leaves. Machine learning models were trained using leaf-level full spectra data, leaf-level Vegetation index (VI) data, and pixel-level full spectra data at four disease progression stages. The results suggest that HSI can detect disease on tomato leaves at pre-symptomatic stages and differentiate bacterial disease spots from abiotic leaf spots.</p>