Stress signaling, response, and adaptive mechanisms in submerged macrophytes under PFASs and warming exposure

Heat stress disturbs cellular homeostasis and alters the fitness of individual organisms. However, it is unclear whether thermal perturbations exacerbate the toxic effects of per- and polyfluorinated alkyl substances (PFASs) on trophic endpoints in freshwater ecosystems. We conducted a mesocosm experiment to investigate the impact of warming and PFASs on the widespread submerged macrophytes (Hydrilla verticillata) at a molecular level. Quantitative and air flow-assisted ionization mass spectrometry imaging results showed that warming significantly increased the accumulation of PFOS (3.53 L/kg) in the submerged leaf tissues. Accumulation of PFASs altered H. verticillata intracellular scavenging enzymes, an effect that may be exacerbated by 4 °C warming. Warming and PFASs influenced photosynthesis, biological rhythms, and ecological stoichiometry, causing a decrease in metabolites linked to the tricarboxylic acid cycle and amino acid metabolism, which compromised nitrogen use efficiency (9.9%-30.4% reduction in nitrogen content, 0.8%-22.8% increase in C:N ratios). Additionally, metabolites are linked to the antioxidant system or cell wall components, with linoleic acid decreasing by 17.1%-82.8% and carbohydrate-related compounds dropping by 52.2%-89.0%. Our modeling analyses revealed that H. verticillata enriched with PFASs could pose secondary risks when consumed by herbivorous fish (Ctenopharyngodon idellus) under warming, potentially affecting food chain dynamics.