Natural restoration has emerged as a prominent approach in recent decades for the rehabilitation of degraded ecosystems globally. However, the specific changes and underlying mechanisms by natural restoration that influence the multifunctionality of karst ecosystems remain poorly understood. In this study, soil, litter, and fine root samples were collected from four chronosequence stages of vegetation restoration—grassland (G), shrubland (SH), shrub-tree land (ST), and forest (F)—within a karst ecosystem in Southwestern China. The aim was to evaluate the impacts of vegetation restoration on ecosystem multifunctionality using an averaging approach. The results demonstrated that the indices of C-cycling functionality, N-cycling functionality, P-cycling functionality, and total ecosystem multifunctionality increased as vegetation restoration progressed, along with plant diversity. The structure of plant, bacterial, and fungal communities varied across different stages of vegetation restoration, exhibiting the highest microbial diversity indices in the SH stage. Additionally, the tightness and complexity of co-occurrence networks of bacteria and fungi increased with advancing vegetation restoration, and higher positive links were observed in fungi than bacteria. The four functional indices were significantly and positively correlated with increasing plant diversity, fine root and litter nutrient contents, fine root biomass, microbial biomass, fungal community, enzyme activities, and soil nutrient contents but not with bacterial and fungal diversities. Furthermore, Random Forest model results revealed that plants exerted a significantly greater influence on ecosystem multifunctionality compared to other factors. It is plausible that plants influence soil microbial biomass, fungal community and co-occurrence networks, enzyme activities, and nutrient levels through the input of root and litter nutrients rather than by altering microbial diversity to enhance karst ecosystem multifunctionality. Therefore, initiatives to increase plant diversity are beneficial for sustainable ecological restoration management in the karst regions of Southwestern China.

Denitrification and nitrification are two pivotal microbial processes relating to N₂O emissions. However, the difference in N₂O emission fluxes and N₂O-producing bacteria between a karst (KA) and non-karst area (NKA) remains unclear. The objective of this study is to compare the differences in soil N₂O emissions, nitrifying bacteria, and denitrifying bacteria during the growth period of rice in KA and NKA, and to explore the mechanisms by which microorganisms and environmental factors drive N₂O emissions. Here, N₂O emission fluxes of paddy fields were collected using the static dark chamber and measured using gas chromatography at KA and NKA in the Maocun Karst Experimental Site in Guilin, China. The nitrifying bacteria (ammonia-oxidizing bacteria, AOB) and denitrifying bacteria (nirK-denitrifier) were determined using real-time PCR and high-throughput sequencing, respectively. Results showed that during the rice growth period, the N₂O emission fluxes in KA was generally lower than that in NKA, with cumulative N₂O emissions of −0.054 and 0.229 kg·hm⁻² in KA and NKA, respectively. The absolute abundance of AOB in KA (8.91 × 10⁶–2.68 × 10⁷ copies·g⁻¹) was significantly higher than that in NKA (1.57 × 10⁶–6.48 × 10⁶ copies·g⁻¹), while the absolute abundance of nirK-denitrifier had no significant difference between the two areas. The composition and diversity of AOB and nirK-denitrifier differed significantly between KA and NKA. Results from partial least squares structural equation modeling (PLS-SEM) indicated that soil properties, carbon sources, and nitrogen sources had positive effects on AOB and nirK-denitrifier, while nirK-denitrifier had a negative effect on N₂O emissions. Partial least squares regression (PLSR) predictions revealed that NO₃⁻-N, SOC, TN, Mg²⁺, Ca²⁺, and pH were the most important factors influencing N₂O emission fluxes. This study highlights the critical role of the typical characteristics of KA soils in reducing N₂O emissions from paddy fields by driving the evolution of AOB and nirK-denitrifier.

Coupled nitrogen (N) and phosphorus (P) cycling in calcareous karst soils, and how it responds to long-term land use and restoration, remains poorly quantified. We compared seven plots on a karst slope platform: three development types (degraded disturbed land, pasture, and fruit orchard) and four restoration types (planted evergreen forest, planted deciduous forest, evergreen–deciduous mixed forest, and naturally restored forest). Gross N transformation rates were measured by ¹⁵N isotope dilution, alongside soil properties, available P, microbial biomass N, and enzyme activities. Conservation plots generally had higher soil organic carbon, total N, microbial biomass N, and enzyme activity than development plots; soil organic carbon peaked under naturally restored forest (57.5 g·kg⁻¹) and was lowest in disturbed land (47.9 g·kg⁻¹). Naturally restored forest also showed the highest gross nitrification and total N mineralization, whereas disturbed land had the weakest ammonification, with negative gross rates pointing to N immobilization. Available P (up to 15.9 mg·kg⁻¹) tracked alkaline phosphatase activity, organic carbon, and total N rather than total P. Across the alkaline range (pH 7.2–7.8), random forest models ranked ammonium and enzyme activity, not pH, as the main predictors of N mineralization. Long-term land use and restoration were thus associated with consistent differences in karst soil N and P supply.