Long-term different fertilization practices restructured the functional carbon pools in a paddy soil through distinct mechanisms

Fertilization is a common management practice for improving soil organic matter (SOM), which is a crucial indicator of soil fertility. Nevertheless, how long-term fertilization affects functional SOM fractions and its mechanisms remain unclear. In this study, a 12-year field experiment, initiated in 2011, was conducted in Xinzhu village, Zhejiang province, China. Four different fertilizer treatments (without any fertilizers application, Control; sole application of inorganic fertilizers, NPK; combined application of NPK with cattle manure, NPKM; and combination of NPK fertilizers and return of rice straw, NPKS) were selected in this field trial. Functional carbon (C) pools, microbial community structure and SOM composition in both topsoil (0-15 cm) and subsoil (15-30 cm) were explored using the physical and chemical fractionation, phospholipid fatty acids (PLFA) analysis and solid-state 13 C-Nuclear Magnetic Resonance Spectrometry (13C 13 C NMR) assays. Compared to the control, the NPK combined with organic amendments had significantly higher organic carbon contents of unprotected pools (cPOM and fPOM) in both topsoil (39-277 %) and subsoil (43.1-91.3 %), respectively. Organic carbon accumulated the most within iPOM (1.99 g kg- 1 soil) and H-d(Silt+Clay) fractions (1.41 g kg- 1 soil) under NPKM in the topsoil, suggesting that physical and chemical protection played the pivotal role after manure addition. Furthermore, NPKM increased the abundance of bacterial and fungal PLFAs in both topsoil (31.4 % and 24.5 %) and subsoil (173 % and 175 %), while NPKS elevated the alkyl/O-alkyl C ratio but reduced the aromaticity in topsoil, compared to inorganic fertilization alone. Partial least squares path modelling showed that fertilization had a direct impact on silt+clay associated C (0.6) in topsoil, POM-C (0.76) in subsoil, and microbial community structure (0.88 and 0.87) in both soil layers. Redundancy analysis revealed that in the NPKM treatment, the key factors affecting functional carbon pools were the total PLFA and the ratio of gram-positive to gram-negative bacterial PLFA, while for the NPKS treatment, it was the alkyl C/O-alkyl C ratio. In conclusion, long-term fertilization had a strong effect on functional carbon fractions through multiple mechanisms in different soil layers, involving changing microbial community and SOM composition.