Response of Ceratophyllum demersum L. and its epiphytic biofilms to 6PPD and 6PPD-Q exposure: Based on metabolomics and microbial community analysis

The emerging contaminant N-(1,3-dimethylbutyl)-N'-phenyl-p-phenylenediamine (6PPD) and its ozone conversion product N-(1,3-dimethylbutyl)-N'-phenyl-p-phenylenediamine quinone (6PPD-Q) pose a threat to aquatic ecosystems. Aquatic animals and plants exhibit vigorous responses at very low ambient concentrations. However, studies of submerged macrophytes, key producers in aquatic ecosystems, are limited and the full extent of their toxic effects and feedback mechanisms is unknown. To investigate the phytotoxicity of 6PPD and 6PPD-Q, we modeled plant responses to abiotic stress using Ceratophyllum demersum L. (C. demersum) as a representative submerged plant. Our findings indicate that 6PPD and 6PPD-Q disrupt physiological and biochemical processes in C. demersum, encompassing growth inhibition, reduction in photosynthetic pigments, induction of oxidative damage, and metabolic alterations. Moreover, unfavorable modifications to biofilms induced were also discernible supported by confocal laser scanning microscopy (CLSM) images and microbial community profiling. More importantly, we found a robust correlation between differentially expressed metabolites (DEMs) and dominant genera, and 6PPD and 6PPD-Q significantly altered their correlation. Overall, our results imply that even though C. demersum is a resilient submerged macrophyte, the toxic effects of 6PPD and 6PPD-Q cannot be disregarded.