Early detection of drought stress is essential for preventing permanent plant damage and minimizing yield loss. This study utilized hyperspectral imaging at the leaf level to visualize drought stress in safflower plants (Carthamus tinctorius L.). Three safflower genotypes, Palenus, A82, and IL-111, were cultivated under three irrigation levels. Stress conditions were simulated by depleting 50%, 70%, and 90% of soil water content, representing unstressed (US), mild stress (MS), and severe stress (SS) conditions, respectively. Hyperspectral images of leaf samples were captured before any visible signs of water scarcity emerged. Classification analysis was performed using the full mean spectral data with partial least squares discriminant analysis, soft independent modeling of class analogy (SIMCA), support vector machines, and artificial neural network (ANN) classifiers. Feature selection methods were applied to extract the most informative wavebands, and ANN was used to build predictive models. Spatial analysis involved pixel-wise classification using both unsupervised (k-means clustering) and supervised (best classifiers) approaches. ANN outperformed other classifiers using the entire spectral data, effectively distinguishing US, MS, and SS classes in the Palenus, A82, and IL-111 genotypes, achieving F1-scores of 92.22%, 96.01%, and 96.47%, respectively. Among the feature selection methods, SIMCA-based features excelled in monitoring stress conditions in the Palenus and A82 genotypes. In supervised spatial analysis, ANN models clearly depicted the progression of stress in leaves across different genotypes. This study demonstrates the potential of hyperspectral imaging to differentiate various levels of drought stress in safflower, an important oilseed crop.
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