Biochar application alters soil metabolites and nitrogen cycle-related microorganisms in a soybean continuous...

Biochar application is a promising practice to enhance soil fertility. However, it is unclear how field-aged biochar affects the soil metabolites and microbial communities in soybean fields. Here, the rhizosphere soil performance after amending with biochar addition rates at 0 (CK), 20 (B20), 40 (B40), and 60 t ha^-1 (B60) was examined via a five-year in-situ field experiment based on a soybean continuous cropping system. Untargeted metabolomics and metagenomics analysis techniques were applied to study the regulatory mechanism of biochar on soybean growth from metabolomics and N cycle microbiology perspectives. We found that the contents of soil total N (TN), available N (Ava N), NH₄⁺-N, and NO₃^--N were significantly increased with biochar addition amounts by 20.0-65.7 %, 3.6-10.7 %, 29.5-57.1 %, and 24.4-46.7 %, respectively. The B20, B40, and B60 triggered 259 (236 were up-regulated and 23 were down-regulated), 236 (220 were up-regulated and 16 were down-regulated), and 299 (264 were up-regulated and 35 were down-regulated) differential metabolites, respectively. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis and topology analysis demonstrated that differential metabolites were highly enriched in seven metabolic pathways such as Oxidative phosphorylation and Benzoxazinoid biosynthesis. Moreover, ten differential metabolites were up-regulated in all three treatments with biochar. Biochar treatments decreased the Nitrospira abundance in soybean rhizosphere soil while increasing Bradyrhizobium abundance significantly in B60. Mantel test revealed that as the biochar addition rate grows, the correlation between Nitrospira and soil properties other than NO₃^--N became stronger. In conclusion, the co-application of biochar with fertilizers is a feasible and effective way to improve soil N supply, even though biochar has undergone field aging. This work offers new insights into the variations in soil metabolites and microbial communities associated with N metabolism processes under biochar addition in soybean continuous cropping soils.