Anthropogenic nitrogen inputs lead to a high ammonium (NH4+)/nitrate (NO3−) ratio in the soil, which restricts hyphal spreading of soil fungi. Access of symbiotic fungi to roots is a prerequisite for plant-fungal interactions. Hyphosphere bacteria protect fungi from environmental stress, yet the impact of hyphosphere bacteria on adaptation of host fungi to NH4+-enriched conditions remains unclear. By developing soil microcosm assays, we report that a plant-symbiotic fungus, Phomopsis liquidambaris, harbors specific hyphosphere bacteria that facilitate hyphal spreading and assist in the root colonization in NH4+-enriched soil. Genetic manipulation, 16S rRNA gene analysis and coinoculation assays revealed that the genus Enterobacter was enriched in the hyphosphere of NH4+-sensitive wild-type compared to NH4+-preferring nitrite reductase‐deficient strain. The representative Enterobacter sp. SZ2-promoted hyphal spreading is only evident in nonsterilized soil. We further identified an increased abundance and diversity of ammonia-oxidizing archaea (AOA) and a synchronously decreased NH4+:NO3− ratio following SZ2 inoculation. Microbial supplementation and inhibitor assays showed that AOA-mediated reduction in NH4+:NO3− ratio is responsible for SZ2-enhanced fungal adaptation to NH4+-enriched conditions. The Ph. liquidambaris-Enterobacter-AOA triple interaction promoted rice growth in NH4+-enriched soil. Our study reveals the essential role of hyphosphere microorganism-based hyphal spreading in plant-fungal symbiosis establishment within nitrogen-affected agroecosystems.
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Supplementary methods, figures, tables and results are provided in the file Supplementary Information. The 16S rRNA and amoAF/amoAR gene sequencing data are available in the NCBI SRA under BioProject PRJNA835591 and PRJNA837065, respectively.
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We would like to acknowledge the National Natural Science Foundation of China (No. 32071638), Jiangsu Funding Program for Excellent and Postdoctoral Talent (2022ZB357), China Postdoctoral Science Foundation (2021M701748), and a project funded by the Priority Academic Program Development (PAPD) of the Jiangsu Higher Education Institutions of China.
The authors declare no competing interests.
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Sun, K., Jiang, HJ., Pan, YT. et al. Hyphosphere microorganisms facilitate hyphal spreading and root colonization of plant symbiotic fungus in ammonium-enriched soil. ISME J 17, 1626–1638 (2023). https://doi.org/10.1038/s41396-023-01476-z