Quantifying physiological trait variation with automated hyperspectral imaging in rice

Advancements in hyperspectral imaging (HSI) together with the establishment of dedicated plant phenotyping facilities worldwide have enabled high-throughput collection of plant spectral images with the aim of inferring target phenotypes. Here, we test the utility of HSI-derived canopy data, which were collected as part of an automated plant phenotyping system, to predict physiological traits in cultivated Asian rice (Oryza sativa). We evaluated 23 genetically diverse rice accessions from two subpopulations under two contrasting nitrogen conditions and measured 14 leaf- and canopy-level parameters to serve as ground-reference observations. HSI-derived data were used to (1) classify treatment groups across multiple vegetative stages using support vector machines (≥ 83% accuracy) and (2) predict leaf-level nitrogen content (N, %, n=88) and carbon to nitrogen ratio (C:N, n=88) with Partial Least Squares Regression (PLSR) following RReliefF wavelength selection (validation: R (2 = )0.797 and RMSEP = 0.264 for N; R (2 = )0.592 and RMSEP = 1.688 for C:N). Results demonstrated that models developed using training data from one rice subpopulation were able to predict N and C:N in the other subpopulation, while models trained on a single treatment group were not able to predict samples from the other treatment. Finally, optimization of PLSR-RReliefF hyperparameters showed that 300-400 wavelengths generally yielded the best model performance with a minimum calibration sample size of 62. Results support the use of canopy-level hyperspectral imaging data to estimate leaf-level N and C:N across diverse rice, and this work highlights the importance of considering calibration set design prior to data collection as well as hyperparameter optimization for model development in future studies.