Biological N fixation (BNF) is an important N input process for terrestrial ecosystems. Long-term N application increases the availability of N, but may also lead to phosphorus (P) deficiency or an imbalance between N and P. Here, we performed a 5-year N application experiment in a subtropical Phyllostachys heterocycla forest in site and a P application experiment in vitro to investigate the effect of N application on the BNF rate and its regulatory factor. The BNF rate, nifH gene, free-living diazotrophic community composition and plant properties were measured. We found that N application suppressed the BNF rate and nifH gene abundance, whereas the BNF rate in soils with added P was significantly higher overall than that in soils without added P. Moreover, we identified a key diazotrophic assembly (Mod#2), primarily comprising Bradyrhizobium, Geobacter, Desulfovibrio, Anaeromyxobacter, and Pseudodesulfovibrio, which explained 77 % of the BNF rate variation. There was a significant positive correlation between the Mod#2 abundance and soil available P, and the random forest results showed that soil available P is the most important factor affecting the Mod#2 abundance. Our findings highlight the importance of soil P availability in regulating the activities of key diazotrophs, and thus increasing P supply may help to promote N accumulation and primary productivity through facilitating the BNF process in forest ecosystems.
Keywords: Biological N fixation; Diazotrophs; Microbial assembly; N application; P application.
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