Precipitation Alleviates Adverse Effects of Nitrogen and Phosphorus Enrichment on Soil Microbial Co-Occurrence Network Complexity and Stability in Karst Shrubland

The karst region is highly ecologically fragile due to its unique geology and poor water and nutrient retention. Despite long-term restoration, vegetation often remains in the secondary shrubland stage. Soil microorganisms play a vital role in maintaining ecosystem functions, but how microbial communities respond to combined water and nitrogen-phosphorus nutrient changes in karst shrubland remains poorly understood. This knowledge gap hinders effective restoration strategies in karst shrublands. Here, the effects of water, nitrogen, and phosphorous additions and their interactions on soil physico-chemical properties, soil microbial abundance, diversity, community composition, and the co-occurrence network were explored. A full factorial experiment (water × nitrogen × phosphorous, each at two levels) was conducted in a karst shrubland with over 20 years of vegetation restoration, with treatments including control, water (+120 mm yr^-1), nitrogen (+20 g N m^-2 yr^-1), phosphorus (+16 g P m^-2 yr^-1), and their four combinations. Our results suggested that water addition significantly increased soil water content and soil microbial abundance but reduced fungal diversity. Nitrogen addition significantly increased soil nitrate nitrogen content and fungal diversity, and fungal diversity showed an increasing trend under phosphorous addition. The addition of nitrogen and phosphorous did not significantly alter the soil microbial community composition, while water addition showed a tendency to change the soil fungal community composition. Network topological properties, robustness, and vulnerability analyses indicated that individual nitrogen or phosphorous additions, as well as their interactions, reduced network complexity and stability. In contrast, water addition alone or in combination with nitrogen and/or phosphorous alleviated these negative effects, and the water and phosphorous interaction exhibited the highest levels of network complexity and stability. Further analysis showed that the soil pH, available phosphorous, ratio of carbon to phosphorous, and ammonium nitrogen were explanatory variables contributing significantly to soil microbial abundance, diversity, community composition, and network complexity. Overall, these findings highlighted the pivotal role of water availability in enhancing soil microbial stability under nutrient enrichment, offering valuable insights into ecological restoration in karst ecosystems.