Dissecting the negative influence mechanisms of microplastics on plant and microbial mediated nitrogen removal in constructed wetlands

The escalating prevalence of microplastics (MPs) in constructed wetlands (CWs) raises concerns about their potential effects on nitrogen (N) removal within these ecosystems. Despite this, systematic investigations into the influence of MPs on N-transformation in CWs, encompassing both phytological and microbial dimensions, are notably lacking. Thus, polyethylene (PE) and polyamide (PA), commonly found in wastewater plants, were studied as typical MPs in CWs over 240 days. Extended accumulation time increased nitrogen inhibition in CWs with PE and PA MPs, notably in PA, reducing TN removal by 7.0 % and 10.5 %. For nitrification process, MPs inhibited the relative abundances (RAs) of genes involved in nitrification and carbon fixation metabolisms in nitrifiers through metagenomic analysis, with PA more significantly inhibiting nitrifying enzyme activities. Regarding denitrification, MPs, especially PA, inhibited the RAs of genes involved in electron production, transport, consumption, ATP production and central carbon metabolism in denitrifiers, and activities of denitrifying enzymes. MPs, particularly PA, were found to elevated ROS level and lactate dehydrogenase release, thus hindering nitrogen transformation. Moreover, MPs caused a reduction in c-di-GMP levels and inhibited genes relevant to biofilm regulation (c-di-GMP signaling and quorum sensing), thereby disturbing extracellular polymeric substance metabolism. Regarding plant systems, MPs, particularly PA, were observed to decrease nitrogen uptake, crucial for N-removal. Additionally, inhibited root activity, reduced photosynthesis, and a compromised antioxidant system also contributed to the diminished N-removal in CWs. Overall, our study systematically reveals how MPs, especially PA, impede nitrogen removal in CWs, contributing to further realize the simultaneous removal of MPs and nitrogen in CWs.