Winter microbial community structure and methane-cycling potential in constructed agricultural wetlands across regions and microhabitats.

Constructed wetlands are widely used to reduce nutrient loading to downstream waters, but they can also emit methane, a potent greenhouse gas. This trade-off between water quality benefits and climate impacts is driven by microbial processes that remain poorly understood in winter. We examined microbial community composition and methane-cycling potential in surface water samples from constructed wetlands in two agricultural regions of Sweden during the winter season, focusing on the effects of emergent vegetation and environmental conditions. Western wetlands, characterized by higher total nitrogen and dissolved oxygen, exhibited significantly greater microbial diversity and more complex co-occurrence networks than eastern wetlands. At the phylum level, Actinobacteriota and Firmicutes were more abundant in the west, while Bacteroidota dominated the east. The effects of emergent vegetation were region-specific: in the west, vegetated zones supported higher diversity and enrichment of plant-associated taxa. Several taxa affiliated with methanotrophs showed higher relative abundance in vegetated zones of the western wetlands, suggesting vegetation may enhance methane oxidation potential in surface waters, even though methane concentrations were similar. Overall, winter microbial networks remained structured, emphasizing the need for integrated microbial and biogeochemical studies to guide wetland design features, such as vegetation and nutrient regimes, that support both methane mitigation and nutrient retention in cold-climate agricultural landscapes.