Potential mechanism of humic acid attenuating toxicity of Pb2+ and Cd2+ in Vallisneria natans

Humic substances are widely present in aquatic environments. Due to the high affinity of humic substances for metals, the interactions have been particularly studied. To assess the effect of humic acid (HA) on submerged macrophytes and biofilms exposed to heavy metal stress, Vallisneria natans was exposed to solutions containing different concentrations of HA (0.5–2.0 mg·L −1 ), Pb 2+ (1 mg·L −1 ) and Cd 2+ (1 mg·L −1 ). Results suggested that HA positively affected the plant growth and alleviated toxicity by complexing with metals. HA increased the accumulation of metals in plant tissues and effectively induced antioxidant responses and protein synthesis. It was also noted that the exposure of HA and metals promoted the abundance and altered the structure of microbial communities in biofilms. Moreover, the positive effects of HA were considered to be related to the expression of related genes resulting from altered DNA methylation levels, which were mainly reflected in the altered type of demethylation. These results demonstrate that HA has a protective effect against heavy metal stress in Vallisneria natans by inducing effective defense mechanisms, altering biofilms and DNA methylation patterns in aquatic ecosystems.