Bioremediation of nitrate in agricultural drainage ditches: Impacts of low-grade weirs on microbiomes and nitrogen cycle gene abundance.

Artificial drainage is essential for the success of modern agriculture, but it can also accelerate the movement of nutrients, especially nitrate, from soil to surrounding and downstream water bodies. Removal of nitrate from agricultural drainage by using controlled drainage systems, such as ditches installed with low-grade weirs, has been shown to help reduce nutrient loading into watersheds. However, the effect of low-grade weirs varies greatly, likely due to the differences in climate, system designs (e.g., hydraulic characteristics), and the resulting variation in microbial structures and functions in the ditch. In this study, we analyzed the temporal and spatial dynamics of microbiomes in a paired ditch system with weir-installed and uninstalled (control) channels over two years by using the 16S rRNA gene amplicon sequencing and the high-throughput quantitative PCR targeting various N cycle-associated genes [the Nitrogen Cycle Evaluation (NiCE) chip]. The installation of the low-grade weir had a significant impact on the microbiome structure and the distribution of denitrifiers. Microbiome structures also differed significantly between the ditch inlets and the outlets. Denitrification functional genes were more abundant in the inlets than in the other locations and in the channel installed with a low-grade weir. Additionally, oxygenic denitrifiers that use nitric oxide dismutase (nod) to produce N and O gases from nitric oxide were detected in the ditch channels, suggesting the occurrence of nitrate removal process that bypasses the production of nitrous oxide (NO). The ditch microbiomes sampled during high-flow seasons (i.e., spring and fall) exhibited greater similarity to each other than microbiomes sampled during low-flow seasons (i.e., summer). Taken together, this study indicates that the low-grade weirs have the potential to foster a more favorable environment for denitrifiers, resulting in an increase in the abundance of denitrification functional genes. These findings could offer valuable insights into system management and optimization strategies.