In a greenhouse experiment, we examined the behavior of biochar in arable soil to demonstrate that these supplements can boost soil carbon storage, as well as to track changes in microbial biomass and identify the microbial communities that use these biochars. In order to ascertain if biochar can consistently alter soil microbial activities, we studied the impact of biochar combination treatments on 16S rRNA gene diversity. In soil treated with biochar, there was a rise in the relative abundance of taxa belonging to the phyla Actinobacteria and Gemmatimonadetes, despite the overall diversity decreasing with biochar addition. According to all of these observations, pyrogenic carbon has a major effect on the composition of the soil microbial community and enriches keystone taxa within the parent soil microbial community. Certain species experienced increases throughout the biochar-amended incubation period, despite the total diversity declining following biochar amendments. The phyla Actinobacteria and Gemmatimonadetes increased in the relative abundance of bacteria in soil treated with biochar, according to DNA sequencing of these species. In summary, these findings show that biochar significantly impacts the constitution and composition of the soil microbial community and enriches important taxa within the parent soil microbial community.

Long-term positioning tests can systematically reveal the evolution characteristics of soil fertility and crop productivity, and reflect the spatiotemporal changes in soil quality and their driving factors. While soil microorganisms mediating nutrient cycling are crucial for maintaining crop productivity and the long-term resilience of agricultural ecosystems, how prolonged use of different fertilization strategies affects their functional capacity remains insufficiently understood. In this study, we applied metagenomic sequencing to investigate how three fertilization treatments, namely (i) N0 receiving only phosphorus (P) and potassium (K) fertilizers, (ii) N250 receiving conventional urea + P and K, and (iii) F250 receiving humic acid urea + P and K, influence soil microbial communities, functional genes related to C and N cycling, and associated soil properties in a long-term field experiment. The F250 treatment significantly increased average annual yields of wheat and maize to 7166.21 kg hm⁻² and 8309.96 kg hm⁻², respectively. These values were 148.66% and 73.47% higher than those under N0, and 8.22% and 11.64% higher than those under N250. Compared with N0, both N250 and F250 signally augmented soil nitrate, ammonium, total nitrogen (TN), and soil organic carbon (SOC), altered microbial community composition, and enhanced the relative abundance of genes engaged in C fixation and methane oxidation. Both treatments also promoted denitrification and dissimilatory nitrate reduction to ammonium (DNRA). Relative to N250, F250 specifically enriched the beneficial bacterial genus Pedobacter, further increased the abundance of the C fixation gene pccA, and markedly upregulated the DNRA gene nrfA. Soil TN and SOC were identified as the key environmental factors regulating microbial community structure and the functional potential of C and N cycling pathways. Collectively, our findings provide a mechanistic understanding of how long-term application of humic acid urea enhances crop productivity by modulating the genetic potential of soil microorganisms in biogeochemical cycles, offering a biological foundation for optimizing fertilization strategies in sustainable agriculture.