Data driven discovery and quantification of hyperspectral leaf reflectance phenotypes across a maize diversity panel

Hyperspectral reflectance data can be collected from large plant populations in a high-throughput manner in both controlled and field environments. The efficacy of using hyperspectral leaf reflectance as a proxy for traits that typically require significant labor and time to collect has been evaluated in a number of studies. Commonly, estimating plant traits using hyperspectral reflectance involves collecting substantial amounts of ground truth data from plant populations, which may not be feasible for many researchers. In this study, we explore the potential of data-driven approaches to analyze hyperspectral reflectance data with little to no ground truth phenotypic measurements. Evaluations were performed using data on the reflectance of 2,151 individual wavelengths of light from the leaves of maize plants harvested from 1,658 field plots of a replicated trial including representatives of 752 maize genotypes from the Wisconsin Diversity Panel. We reduced the dimensionality of this dataset using an autoencoder neural network and principal component analyses, producing 10 latent variables and principal components, respectively. A subset of these principal components and latent variables demonstrated significant repeatability, indicating that a substantial proportion of the total variance in these variables was explained by genetic factors. Moreover, correlations were observed between variables derived from the autoencoder network and principal components with molecular traits. Notably, the most relevant latent variable (LV8) showed a much stronger correlation with chlorophyll content (R2 = 0.59) compared to the most correlated principal component (PC2; R2 = 0.31). Furthermore, one latent variable exhibited modestly better performance than a partial least squares regression model in estimating leaf chlorophyll content (PLSR; R2 = 0.58, LV8; R2 = 0.59). A number of genetic markers in the maize genome were significantly correlated with variation in different latent variables in genome wide association studies. In a number of cases, significant signals in genome wide association studies were adjacent to genes with plausible links to traits expected to influence leaf hyperspectral reflectance patterns. ### Competing Interest Statement James C. Schnable has equity interests in Data2Bio; Dryland Genetics; and EnGeniousAg. He is a member of the scientific advisory board of GeneSeek and currently serves as a guest editor for The Plant Cell. The authors declare no other conflicts of interest.