Soil phosphorus drives variation in diazotrophic communities in a subtropical nitrogen-rich forest

Although nitrogen (N)-fixing bacteria (diazotrophs) play a vital role in the biosphere's N cycle. However, large uncertainty exists regarding how atmospheric N deposition regulates biological N fixation (BNF) in soils and which diazotrophic taxa are responsible for this process. We targeted the nifH gene to investigate the effects of long-term N addition on the abundance and composition of diazotrophic communities in subtropical N-rich forests, using real-time quantitative polymerase chain reaction (q-PCR) and MiSeq sequencing. We conducted a field trial for seven years and included three N treatment levels (+0, +50, +150 kg N ha-1 yr � 1; CK, LN, HN). With increasing N addition, the average total P concentration in the whole soil profile and the topsoil P availability decreased significantly, further reducing nitrogenase activity and nifH abundance significantly, while increasing the diversity of the diazotrophic community. After seven years of N addition, the relative abundance of uncultured Cyanobacteria significantly decreased, while those of Rhizobiales and Desulfobulbaceae significantly increased. The diazotrophic communities transitioned from free-living N fixers (uncultured Cyanobacteriadominated) to symbiotic N fixers (Rhizobiales-dominated; HN treatment). The relative abundance of heterotrophic diazotrophic bacteria and the occasional diazotrophic species increased, resulting in diazotrophic bacteria still maintaining high competitiveness in the N-rich and/or P-limited soils. Proteobacteria (e.g., Rhizobiales) had the highest potential to participate in microbial mechanisms triggered by P scarcity. Variation partitioning analyses showed that diazotrophic community composition and diversity were mainly correlated with the soil P availability, with soil AP having the highest individual effect. Overall, this study will broaden our understanding of N fixation mechanisms and the diazotrophic community variation in N-rich forests, while also highlighting soil P as a key factor driving changes in diazotrophic communities.

Automated summary

By targeting the nifH gene, this study investigated the effects of long-term nitrogen addition on the abundance and composition of diazotrophic communities in subtropical N-rich forests. The results showed that increasing nitrogen addition decreased nitrogenase activity and nifH abundance, but increased the diversity of diazotrophic communities. Additionally, the composition of diazotrophic communities shifted from free-living nitrogen fixers to symbiotic nitrogen fixers.