Long-Term Biogas Slurry Application Drives Two-Phase Succession in Sugarcane Field Soil Ecosystems: From Microbial Community Disturbance to Functional Restructuring

Promoting the agricultural recycling of biogas slurry (BS) is crucial for sustainable development, yet its long-term ecological impacts remain unclear. Through a multi-year field trial in a sugarcane system, this study examined the effects of BS application (0, 3, and 6 years) on the soil properties, bacterial communities, and functional genes for C, N, P, and S cycling. The results revealed distinct two-phase patterns of changes in soil properties, microbial communities, and functional genes. Short-term (3-year) application induced a “disturbance” phase, characterized by significant acidification (pH decreased by 17.91%), a surge in nitrate-N (increased by 757.27%), and a transient decline in bacterial richness. Long-term (6-year) application drove a “functional restructuring” phase, reversing acidification and significantly increasing soil organic matter (29.05%) and total nitrogen (TN) (20.81%). Bacterial richness recovered, and community composition distinctively restructured. Functional gene analysis revealed shifts in gene abundance that transitioned from high abundance in the short term to a new balance favoring processes like N fixation. Co-occurrence network analysis indicated that this functional shift was associated with core microbial modules (e.g., Firmicutes) and changes in soil pH and SOM. This study suggests that, although short-term application causes significant adjustments, sustained and appropriate BS application can ultimately enhance soil fertility and promote a functionally reorganized state by reshaping microbial interaction networks. It presents a microbial ecological basis for the safe and sustainable use of BS in circular agriculture.