Comammox Nitrospira play an active role in nitrification of agricultural soils amended with nitrogen fertilizers.

The recent discovery of complete ammonia oxidizers (comammox Nitrospira) challenged the paradigm of the two-step nitrification mediated by two distinct groups of nitrifiers, and raised fundamental questions regarding their niche specialization and relative contribution to nitrification in agricultural soils. Previous studies suggest that comammox Nitrospira have an oligotrophic lifestyle and would outcompete canonical ammonia oxidizers (ammonia-oxidizing bacteria and ammonia-oxidizing archaea) under ammonia-limited conditions. Here, we demonstrated that comammox Nitrospira clade A were significantly more abundant than canonical ammonia oxidizers and 13CO 2 -DNA-stable isotope probing revealed that comammox Nitrospira clade A incorporated 13CO 2 into their genomes in fertilized agricultural soils during the microcosm incubation. Phylogenetic analysis of the amoA gene revealed that 13CO 2 -labelled comammox Nitrospira clade A belonged to the Nitrospira inopinata -related cluster and a new cluster that was distinct from the known comammox isolates. These results demonstrated the potential important role of comammox Nitrospira in autotrophic ammonia oxidation in agricultural soils amended with nitrogen fertilizers and their lifestyle may be not strictly restricted to oligotrophic habitats. There is a potential contribution of comammox Nitrospira to soil nitrification, which calls re-evaluation of the microbial nitrogen cycling processes and the subsequent impacts on agriculture and the environment. • Comammox Nitrospira clade A played a potential role in nitrification of fertilized agricultural soils. • The N fertilizer input significantly increased the abundances of comammox Nitrospira clade A, AOA and AOB. • Acetylene significantly impeded comammox clade A, AOA and AOB, but 1-octyne only inhibited AOB abundance. • Comammox Nitrospira clade A incorporated 13CO 2 into their genomes during the incubation. • A new cluster of comammox Nitrospira may play an important role in nitrification of fertilized soils. [ABSTRACT FROM AUTHOR]