Soil aggregates, which are the basic units of soil structure, play an important role in the carbon cycle of ecosystems. The pore characteristics of aggregates influence soil organic carbon sequestration. However, studies on SOC mechanisms in aggregates have been limited to Mollisols. This study was conducted as a long-term experiment established in 2004 with a corn-soybean rotation in Mollisols. There are three treatments, including rotary tillage without straw return (conventional tillage, CT), subsoiling without straw return (reduced tillage, RT), and no tillage with straw return (NT). The soil pore size distribution, shape parameters, extracellular enzymes activity, and carbon mineralization were measured. The results showed that 15-year no tillage and reduced tillage increased the total porosity and proportion of larger pores, but significantly decreased the proportion of smaller pores in situ soil columns. Conventional tillage exhibited the most complex pores because of the highest pore fractal dimension (2.75-2.90), anisotropy (0.366-0.516), and the lowest sphericity (5.1-28.7). As for the soil columns filled with > 2 mm aggregates, reduced tillage significantly increased the pore connectivity by 3.02-3.62 %, whereas no tillage had no effect. The structural equation modelling indicated that in soil columns filled with > 2 mm aggregates, pore shape parameters, particularly connectivity and anisotropy, positively influenced the activities of beta-glucosidase and beta-xylosidase directly, and positively affected soil carbon mineralization by influencing extracellular enzymes activity indirectly. The findings emphasize the importance of pore shape parameters effect on soil carbon sequestration, and will be helpful in comprehending the microscopic mechanisms of soil carbon sequestration in > 2 mm aggregates.

Soil freeze-thaw cycles (FTCs) are common in temperate agricultural ecosystems during the non-growing season and are progressively influenced by climate change. The impact of these cycles on soil microbial communities, crucial for ecosystem functioning, varies under different agricultural management practices. Here, we investigated the dynamic changes in soil microbial communities in a Mollisol during seasonal FTCs and examined the effects of stover mulching and nitrogen fertilization. We revealed distinct responses between bacterial and fungal communities. The dominant bacterial phyla reacted differently to FTCs: for example, Proteobacteria responded opportunistically, Actinobacteria, Acidobacteria, Choroflexi and Gemmatimonadetes responded sensitively, and Saccharibacteria exhibited a tolerance response. In contrast, the fungal community composition remained relatively stable during FTCs, except for a decline in Glomeromycota. Certain bacterial OTUs acted as sensitive indicators of FTCs, forming keystone modules in the network that are closely linked to soil carbon, nitrogen content and potential functions. Additionally, neither stover mulching nor nitrogen fertilization significantly influenced microbial richness, diversity and potential functions. However, over time, more indicator species specific to these agricultural practices began to emerge within the networks and gradually occupied the central positions. Furthermore, our findings suggest that farming practices, by introducing keystone microbes and changing interspecies interactions (even without changing microbial richness and diversity), can enhance microbial community stability against FTC disturbances. Specifically, higher nitrogen input with stover removal promotes fungal stability during soil freezing, while lower nitrogen levels increase bacterial stability during soil thawing. Considering the fungal tolerance to FTCs, we recommend reducing nitrogen input for manipulating bacterial interactions, thereby enhancing overall microbial resilience to seasonal FTCs. In summary, our research reveals that microbial responses to seasonal FTCs are reshaped through land management to support ecosystem functions under environmental stress amid climate change.