Close-range hyperspectral image analysis for the early detection of stress responses in individual plants in a high-throughput phenotyping platform

The potential of close-range hyperspectral imaging (HSI) as a tool for detecting early drought stress responses in plants grown in a high-throughput plant phenotyping platform (HTPPP) was explored. Reflectance spectra from leaves in close-range imaging are highly influenced by plant geometry and its specific alignment towards the imaging system. This induces high uninformative variability in the recorded signals, whereas the spectral signature informing on plant biological traits remains undisclosed. A linear reflectance model that describes the effect of the distance and orientation of each pixel of a plant with respect to the imaging system was applied. By solving this model for the linear coefficients, the spectra were corrected for the uninformative illumination effects. This approach, however, was constrained by the requirement of a reference spectrum, which was difficult to obtain. As an alternative, the standard normal variate (SNV) normalisation method was applied to reduce this uninformative variability. Once the envisioned illumination effects were eliminated, the remaining differences in plant spectra were assumed to be related to changes in plant traits. To distinguish the stress-related phenomena from regular growth dynamics, a spectral analysis procedure was developed based on clustering, a supervised band selection, and a direct calculation of a spectral similarity measure against a reference. To test the significance of the discrimination between healthy and stressed plants, a statistical test was conducted using a one-way analysis of variance (ANOVA) technique. The proposed analysis techniques was validated with HSI data of maize plants (Zea mays L.) acquired in a HTPPP for early detection of drought stress in maize plant. Results showed that the pre-processing of reflectance spectra with the SNV effectively reduces the variability due to the expected illumination effects. The proposed spectral analysis method on the normalized spectra successfully detected drought stress from the third day of drought induction, confirming the potential of HSI for drought stress detection studies and further supporting its adoption in HTPPP

Analysis of hyperspectral images for detection of drought stress and recovery in maize plants in a high-throughput phenotyping platform

The study of physiological processes resulting from water-limited conditions in crops is essential for the selection of drought-tolerant genotypes and the functional analysis of related genes. A promising, non-invasive technique for plant trait analysis is close-range hyperspectral imaging (HSI), which has great potential for the early detection of plant responses to water deficit stress. In this work, a data analysis method is described that, unlike vegetation indices, the present method applies spectral similarity on selected bands with high discriminative information, while requiring a careful treatment of uninformative illumination effects. The latter issue is solved by a standard normal variate (SNV) normalization that removes linear effects and a supervised clustering approach to remove pixels that exhibit nonlinear multiple scattering effects. On the remaining pixels, the stress-related dynamics is quantified by a spectral analysis procedure that involves a supervised band selection procedure and a spectral similarity measure against well-watered control plants. The proposed method was validated by a large-scale study of water-stress and recovery of maize plants in a high-throughput plant phenotyping platform. The results showed that the analysis method allows for an early detection of drought stress responses and of recovery effects shortly after re-watering