Micro(nano)plastic size and concentration co-differentiate nitrogen transformation, microbiota dynamics, and assembly patterns in constructed wetlands

Micro/nano-sized plastics (MPs/NPs) existing in wastewater system are the potential threats to nitrogen (N) biotransformation. Constructed wetlands (CWs) as wastewater treatment systems are considered the important barriers preventing MPs/NPs from entering the open water. However, little is known about how the accumulation of MPs/NPs affects microbial N transformation, dynamics, assembly, and metabolism of wetland microbiota. Herein, we constructed 12 wetland systems to address the above knowledge gaps over 300-day exposure to different sizes (3 mm - 60 nm) and concentrations (10 - 1000 μg/L) of MPs/NPs. The results showed that MPs/NPs accumulation caused decrease in NH 4 + -N removal (by 7.6% - 71.2%) and microbial diversity and intriguingly altered microbiota composition (especially in the high-concentration groups) without damage on the high removal efficiency of NO 3 − -N and NO 2 − -N (66.2% - 99.8%) in all except for the nano-sized plastic-exposed wetlands. Moreover, MPs/NPs exposure induced shift in the strengths of non-random species aggregation and segregation patterns co-differentiated by the size and concentration of MPs/NPs, and MPs/NPs accumulation created size-differentiated alternative niches for nitrogen-transforming bacteria, e.g., canonical nitrifiers ( Nitrospira and Nitrosomonas ) and denitrifiers ( Thauera, Comamonas , and Aquabacterium ), which were enriched in MPs groups where denitrifying enzyme-coding genes were also enriched, suggesting potential positive impact of larger plastics on denitrification. Our study highlights MPs/NPs-induced divergence in microbiota dynamics and nitrogen transformation in CWs, and provides important insights into how microbiota structurally and functionally respond to long-term MPs/NPs disturbance.