High Ratio of Manure Substitution Enhanced Soil Organic Carbon Storage via Increasing Particulate Organic Carbon and Nutrient Availability

Replacing partial chemical fertilizers with organic fertilizer can increase organic carbon input, change soil nutrient stoichiometry and microbial metabolism, and then affect soil organic carbon (SOC) storage. A 6-year field experiment was used to explore the mechanism of SOC storage under different ratios of manure substitution in northeast China, with treatments including chemical fertilizer application alone (nitrogen, phosphorus, and potassium, NPK) and replacing 1/4 (1/4M), 2/4 (2/4M), 3/4 (3/4M), and 4/4 (4/4M) of chemical fertilizer N with manure N. Soil nutrients, enzymatic activity, and SOC fractions were analyzed to evaluate the effect of different manure substitution ratios on SOC storage. A high ratio of manure substitution (>1/4) significantly increased soil total N, total P, total K, and available nutrients (NO3−-N, available P, and available K), and the 4/4M greatly decreased the C/N ratio compared to the NPK. Manure incorporation increased microbial biomass carbon (MBC) by 18.3–53.0%. Treatments with 50%, 75%, and 100% manure substitution (2/4M, 3/4M, and 4/4M) enhanced bacterial necromass carbon (BNC), fungal necromass carbon (FNC), and total microbial necromass carbon (MNC) by 31.9–63.5%, 25.5–107.1%, and 27.4–94.2%, respectively, compared to the NPK treatment. Notably, the increase in FNC was greater than that of BNC as the manure substitution ratio increased. The increasing manure substitution significantly enhanced particulate organic C (POC) and total SOC but did not affect mineral-associated organic C (MAOC). High soil N and P supplies decreased leucine aminopeptidases (LAPs) and alkaline phosphatase activities but increased the activity ratio of β-glucosidase (BG)/(N-acetyl-glucosaminidase (NAG) + LAP). Treatments with 25% manure substitution (1/4M) maintained maize and soybean yield, but with increasing manure rate, the maize yield decreased gradually. Overall, the high ratio of manure substitution enhanced SOC storage via increasing POC and MNC, and decreasing the decomposition potential of manure C and soil C resulting from low N- and P-requiring enzyme activities under high nutrient supplies. This study provides empirical evidence that the rational substitution of chemical fertilizers with manure is an effective measure to improve the availability of nutrients, and its effect on increasing crop yields still needs to be continuously observed, which is still a beneficial choice for enhancing black soil fertility.