Inter-Row Grassing Reshapes Nitrogen Cycling in Peach Orchards by Influencing Microbial Pathways in the Rhizosphere

Traditional clean tillage in peach orchards leads to soil degradation and nitrogen (N) loss. While inter-row grassing can optimize N cycling, the specific rhizosphere microbial mechanisms involved have not been fully understood. This study investigated how different inter-row grassing modes influence N availability through microbial communities in a peach orchard. The experiment included a monoculture of <i>Trifolium repens</i> L. (Tr), a monoculture of <i>Lolium perenne</i> L. (Pr), their mixture (TPr), and clean tillage (CK). By combining soil physicochemical analyses, metagenomic sequencing, functional gene quantification, and multivariate statistics, the study systematically examined the impacts of inter-row grassing modes on soil N cycling. The results showed that inter-row grassing modes played a significant role in reshaping N processes. Pr enhanced mineralization and nitrification, increasing inorganic N through specific genes (<i>amoA</i>, <i>hao</i>). Tr, on the other hand, promoted diazotrophs (<i>Bradyrhizobium</i>) and dissimilatory nitrate-reducing bacteria, enhancing biological N fixation and retention. TPr combined these benefits, leading to enhanced nitrification, increased labile carbon, and elevated enzyme activities, creating a complex microbe-gene network that mediated nitrification and denitrification. Overall, inter-row grassing modulates rhizosphere functions by enhancing N cycling through a "carbon input-microbial regulation" mechanism, offering an effective strategy for improving N use efficiency and promoting sustainable orchard management.