Changes in soil physicochemical properties and bacterial community composition were investigated six years after biochar amendment at 0%, 4%, 8% and 12% (w/w), which were coded as C0, C1, C2 and C3, respectively. Results showed that some soil characteristics were sustainable, as they were still affected by biochar addition after six years. Compared to the control, biochar-treated soils had higher pH, total carbon (TC), C/N, total nitrogen (TN), available phosphorus (AP) and available potassium (AK). Soil pH, C/N and the content of TC, TN and AK all increased along with the increase of biochar dosage. The results of Illumina MiSeq sequencing demonstrated that biochar enhanced soil bacteria diversity and modified the community composition over time. The relative abundance of Nitrospirae and Verrucomicrobia phylum increased but that of Acidobacteria phylum decreased significantly in biochar amended soils. The addition of biochar also enriched some bacterial genera, such as uncultured Nitrosomonadace, uncultured Gemmatimonadac, uncultured Nitrospiraceae and Magnetovibrio. In particular, the relative abundance of uncultured Nitrospiraceae was enhanced by 16.9%, 42.8% and 73.6% in C1, C2 and C3, respectively, compared to C0. Biochar has a potential role in enhancing the abundance of bacteria involved in N cycling. Soil pH, TC, TN, TK and AK, were closely related to alterations in the composition of the soil bacterial community. Meanwhile, these soil properties were significantly influenced by biochar amendment, which indicates that biochar affected the soil microbial community indirectly by altering the soil characteristics in the long term.

Crops assemble and rely on rhizosphere-associated microbiomes for plant nutrition, which is crucial to their productivity. Historically, excessive nitrogen fertilization did not result in continuously increasing yields but rather caused environmental issues. A comprehensive understanding should be developed regarding the ways in which crops shape rhizosphere-associated microbiomes under conditions of increased nitrogen fertilization. In this study, we applied 16S and 18S ribosomal RNA gene profiling to characterize bacterial and fungal communities in bulk and rhizosphere soil of rice subjected to three levels of nitrogen fertilization for 5 years. Soil biochemical properties were characterized, and carbon-, nitrogen-, and phosphorus-related soil enzyme activities were investigated, by assays. Increasing nitrogen fertilization led to a decreasing trend in the variation of microbial community structures and demonstrated a more definite influence on fungal rather than bacterial community compositions and functions. Changes in the level of nitrogen fertilization significantly affected chemical properties such as soil pH, nutrient content, and microbial biomass levels in both rhizosphere and bulk soil. Soil enzyme activity levels varied substantially across nitrogen fertilization intensities and correlated more with the fungal than with the bacterial community. Our results indicated that increased nitrogen input drives alterations in the structures and functions of microbial communities, properties of soil carbon, nitrogen, and phosphorus, as well as enzyme activities. These results provide novel insights into the associations among increased nitrogen input, changes in biochemical properties, and shifts in microbial communities in the rhizosphere of agriculturally intensive ecosystems.

Soil fungal microbiomes facilitate a range of beneficial functions for their host plants, and rhizosphere fungal community composition, richness, and diversity affect plant growth and development, and crop yield. Therefore, exploring the community structure and assembly of the rhizosphere fungal microbiome and its relationship with soil biochemical properties are fundamental to elucidating how rice plants benefit from their fungal symbionts. In this study, soil samples were collected at seedling, tillering, heading, and ripening stages of rice subjected to three levels of nitrogen fertilization. Plant growth demonstrates a substantial influence on fungal community composition and diversity. From the tillering to the ripening stage, the fungal communities were governed by homogenizing dispersal and dispersal limitation. The prevalence of Glomeromycota, the beneficial fungi, was considerably higher during the heading stage compared to the three other growth stages. This increase in abundance was strongly associated with increased levels of soil nutrients and enhanced activity of nitrogen acquisition enzymes. This may be a strategy developed by rice grown in flooded soil to recruit beneficial fungi in the rhizosphere to meet high nitrogen demands. Our study findings contribute to elucidating the influence of plant development and nitrogen fertilization on the structure and composition of the fungal community as well as its relationship with soil key soil nutrient content and nitrogen-related enzyme activities. They also illustrate how a shift in the fungal community mediates and reflects the effects of nitrogen fertilization input in rice agroecosystems. These findings provide new insights into the effects of changes in nitrogen application in rice rhizosphere at different growth stages on fungal communities and soil biochemical characteristics.