Identifying Plant Stress Responses to Roxarsone in Soybean Root Exudates: New Insights from Two-Dimensional...

Roxarsone (ROX) is an organoarsenic feed additive of increasing interest used in the poultry industry. Soybean responses to ROX stress were investigated in root exudates (REs) using two-dimensional correlation spectroscopy (2D-COS) with fluorescence and Fourier transform infrared spectra. Environmentally relevant ROX concentrations caused negligible toxicity to crop growth and photosynthesis activity but blackened soybean roots at high concentrations. 2D-COS analysis revealed that the protein-like fluorophore and C═C and C═O, aliphatic OH, and polysaccharide C-O-H moieties in soybean REs were most sensitive to ROX stress. Heterospectral 2D-COS results suggested that aromatic, amide I, quinone, ketone, and aliphatic functional groups were the foundational components of protein-like and short-wavelength excited humic-like fluorophores in soybean REs. Carboxyl and phenolic moieties were related to the long-wavelength excited humic-like fluorophore. Overall, 2D-COS combined with molecular-based spectral analysis of REs provided an innovative approach to characterize the physiological responses of crops to contaminants at sublethal levels.