Long term combined fertilization and soil aggregate size on the denitrification and community of denitrifiers

Abstract

Balanced fertilization strategies that combine the use of organic and inorganic fertilizers have been adopted to increase soil fertility and crop yield and to safeguard environment. However, knowledge on the influence of these fertilization strategies on microbe-mediated nitrogen cycling in the agricultural ecosystems remains limited. We analyzed the denitrifiers with nitrite reductase genes (nirK and nirS) in three soil aggregate classes (large macroaggregates, >2 mm; small macroaggregates, 0.25–2 mm; and microaggregates, <0.25 mm) in response to long-term application of inorganic fertilizers (NPK), NPK plus straw (NPKS), and NPK plus manure (NPKM) in the fluvo-aquic soil of North China Plain. Both fertilization and aggregate size had significant effects on the nirK and nirS gene abundances and soil potential denitrification activity (PDA). PDA in the combined fertilizer-treated soils (NPKS and NPKM treatments) was two times more than that in inorganic NPK-treated soil. PDA dramatically increased in small macroaggregates and microaggregates of NPKS treatment. The nirS gene abundance was significantly higher than that of the nirK gene abundance, and NPKS and NPKM treatments increased the nirS to nirK ratio in bulk soils and in microaggregates compared with NPK treatment and control. Correlation analysis and automatic linear modeling revealed a primary correlation between PDA and abundance of nirS-type denitrifiers. Soil properties, such as SOC, pH and TN, significantly affected the community structure of nirK- and nirS-type denitrifiers while only aggregate size significantly influenced the composition of nirK-type denitrifiers. Taken together, our results suggest that long-term combined fertilization increased the denitrification potential in small macroaggregate and microaggregate classes, and the increased gene abundance and the shift community structures of nirS- and nirK-type denitrifiers were potentially responsible for an increase in the promoted denitrification.