Changes of the denitrifying communities in a multi-stage free water surface constructed wetland

Microorganisms play crucial roles in the nitrogen removal processes of wetlands. However, the key functional genes and microbes related to the nitrogen removal remain largely unknown in the free water surface constructed wetland (FWS CW). Here we studied the abundances of denitrifiers by targeting the key functional genes (nirS, nirK and nosZ) and investigated the community compositions of denitrifiers and their correlations with the abiotic variables in a FWS CW. The increase of nosZ/(nirS + nirK) and nirS/nirK ratios in the outlet indicated a shift of denitrifiers' communities which tended to release less nitrous oxide at the genetic potential level. The denitrifiers dominated the bacterial community which also remarkably changed from the inlet to the outlet. PICRUSt analysis revealed that the denitrifiers contributed to 39.1% of the nitrogen metabolism, 38.9% of the amino acid metabolism and 25.6% of the amino acid related enzymes. Four bacterial genera including Hydrogenophaga, Hylemonella, Aquabacterium and Cellvibrio were detected as the putative keystone denitrifiers. The abundance (nirS, nirK and nosZ) and the relative abundance of putative keystone denitrifiers were significantly correlated with total organic carbon, oxidation-reduction potential and C/N ratio, which could be regarded as the determinants for the denitrification process in the free water.