Combined application of organic manure and chemical fertilizers stabilizes soil N-cycling microflora

·The abundance of N-cycling genes differently responded to NPK application.·Chemical NPK application greatly altered the N-cycling microbial community structure.·Soil acidification was the main driver for the variation in the N-cycling microbial community.·Manure addition was beneficial for stabilizing the N-cycling microbial community.Straw and manure are widely applied to agricultural systems,and greatly shape soil N-cycling microflora.However,we still lack a comprehensive understanding of how these organic materials structure soil N-cycling microbial communities.In this study,metagenomic analysis was performed to investigate the compositional variation in N-cycling microbial communities in a 30-year long-term experiment under five fertilization regimes:no fertilization(Control),chemical fertilization only(NPK),and NPK with wheat straw(NPK+HS),pig manure(NPK+PM),and cow manure(NPK+CM).Long-term NPK application differentially changed N-cycling gene abundance and greatly altered N-cycling microbial community structure.NPK+HS resulted in a similar pattern to NPK in terms of gene abundance and community structure.However,NPK+PM and NPK+CM significantly increased most genes and resulted in a community similar to that of the Control.Further analysis revealed that serious soil acidification caused by long-term NPK fertilization was a major factor for the variation in N-cycling microbial communities.The addition of alkaline manure,rather than wheat straw,stabilized the N-cycling microbial community structure presumably by alleviating soil acidification.These results revealed the strong impact of soil acidification on microbial N-cycling communities and illustrated the possibility of resolving nitrogen-related environmental problems by manipulating pH in acidified agricultural soils.