Non-Destructive Detection of Heat Stress in Tobacco Plants Using Visible-Near-Infrared Spectroscopy and Aquaphotomics Approach

Non-destructive estimation of high-temperature stress effects on tobacco plants is crucial for both scientific research and practical applications. Normalized difference vegetation index (NDVI), chlorophyll index, and spectra in the range of 900–1700 nm of Burley, Oriental, and Virginia tobacco plants under control and high-temperature stress conditions were measured using portable instruments. NDVI and chlorophyll index measurements indicate that young leaves of all tobacco types are tolerant to high temperatures. In contrast, the older leaves (the fifth leaf) showed increased sensitivity to heat stress. The chlorophyll content of these leaves decreased by 40 to 60% after five days of stress, and by the seventh day, the reduction reached 80% or more in all plants. The vegetative index of the fifth leaf also decreased on the seventh day of stress in all tobacco types. Differences in near-infrared spectra were observed between control, stressed, and recovered plants, as well as among different stress days, and among tobacco lines. The most significant differences were in the 1300–1500 nm range. The first characterization of heat-induced changes in the molecular structure of water in tobacco leaves using an aquaphotomics approach was conducted. Models for determining days of high-temperature treatment based on near-infrared spectra achieved a standard error of cross-validation (SECV) from 0.49 to 0.62 days. The total accuracy of the Soft Independent Modeling of Class Analogy (SIMCA) classification models of control, stressed, and recovered plants ranged from 91.0 to 93.6% using leaves’ spectra of the first five days of high-temperature stress, and from 90.7 to 97.7% using spectra of only the fifth leaf. Similar accuracy was obtained using Partial Least Squares–Discriminant Analysis (PLS-DA). Near-infrared spectroscopy and aquaphotomics can be used as a fast and non-destructive approach for early detection of stress and additional tools for investigating high-temperature tolerance in tobacco plants.