Soil carbon storage and accessibility drive microbial carbon use efficiency by regulating microbial diversity and key taxa in intercropping ecosystems

Intercropping is a powerful practice to alter the allocation of photosynthetic carbon (C) to belowground ecosystems via promotion of diversified plant communities. The feedback of soil C stability to intercropping is controlled by microbial C use efficiency (CUE). Despite its significance, there is currently insufficient evidence to decipher how soil microbial CUE reacts to intercropping. By combining a 10-year-long intercropping experiment with a substrate-independent ¹⁸O-H₂O labelling approach and high-throughput sequencing, we elucidated the performance of intercropping on soil C pool and microbial metabolic traits as well as their relationships with soil microbial communities. Compared with monoculture, maize intercropping with peanut and soybean significantly increased soil C storage, soil mineral-associated organic C (MAOC), soil dissolved organic (DOC), and soil microbial biomass (MBC) contents at maize four growth stages. Soil microbial CUE increased significantly, especially at maize flowering and mature stages, as a consequence of enhanced microbial growth and biomass turnover rate after maize intercropping with peanut and soybean. Soil C storage and accessibility indicators (e.g., MAOC, DOC, and MBC contents) could significantly predict the changes of soil microbial diversity and core taxa. Meanwhile, the beta-diversity (community composition) of soil bacteria, fungi, saprotroph and protists, as well as rare fungal taxa were positively correlated with soil microbial CUE, and these indicators showed a high prediction of the microbial CUE. Soil C storage and accessibility indicators directly and indirectly influenced soil microbial CUE by regulating microbial diversity and key taxa. Soil microbial diversity and core taxa directly and indirectly influenced microbial CUE by mediating microbial respiration, growth, biomass, and enzyme activity, which mediated by soil C storage and accessibility. These findings provide an evidence for the associations between microbial diversity, CUE, and soil C stability, highlighting the importance of intercropping-driven soil microbiome to enhance soil microbial CUE.