Unveiling microbial community and function involved in anammox in paddy vadose under groundwater irrigation

The extensive application of nitrogen fertilizer in intensive irrigation areas poses a potential threat to groundwater. Given that the vadose zone acts as a buffer zone for the underground entry of surface pollutants, an in-depth understanding of its microbial community structure and function was crucial for controlling groundwater nitrogen pollution. In this study, soil samples from paddy vadose under groundwater irrigation with different depths (G1: 6.8 m, G2: 13.7 m, G3: 15.6 m, and G4: 17.8 m) were collected to unravel the differences in microbial community structure and function at different vadose depths (0-250 cm), as well as their relationship with soil properties. Results showed some differences among soil physicochemical factors under groundwater irrigation with different depths and that some electron acceptors were more abundant than others under deep groundwater irrigation (G2-G4). Remarkable differences in microbial communities under shallow- and deep-groundwater irrigation were found. The high abundances of anammox bacteria Candidatus_Brocadia in G2 and G3 indicated that deep groundwater irrigation was beneficial to its enrichment. Iron-reducing bacteria Anaeromyxobacter and sulfate-reducing bacteria Desulfovibrio were widely distributed in vadose zone and possessed the potential for anammox coupled with Fe(III)/sulfate reduction. Norank_f_Gemmatimonadaceae had nitrate- and vanadium-reducing abilities and could participate in anammox in vadose zone. Dissimilatory nitrate reduction to ammonia (DNRA) bacteria Geobacter facilitated Fe(II)-driven DNRA and thus provided electron donors and acceptors to anammox bacteria. Soil nutrients and electron donors/acceptors played important roles in shaping microbial community structure at phylum and genus levels. Microorganisms in vadose zone under groundwater irrigation showed good material/energy metabolism levels. Deep groundwater irrigation was conducive to the occurrence of anammox coupled with multi-electron acceptors. Our findings highlight the importance of understanding the structure and function of microbial communities in paddy vadose under groundwater irrigation and reveal the potential role of indigenous microorganisms in in-situ nitrogen removal.