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In the artificial cultivation of truffles, ectomycorrhizal colonization level, host plant quality, and the associated microbes in the rhizosphere soil are vitally important. To explore the effects of nitric oxide (NO) and phosphorus (P) stress on the early symbiosis of truffles and host plants, different concentrations of exogenous NO donor sodium nitroprusside (SNP) and P were applied to  Carya illinoinensis  seedlings inoculated with the Chinese black truffle ( Tuber indicum ). The growth of  T. indicum -mycorrhized seedlings and their mycorrhizal colonization rate were investigated. Additionally, the denitrifying bacterial community harboring NO reductase ( norB ) genes and the fungal community in the rhizosphere of the host were analyzed by high-throughput sequencing. The results showed that the colonization rate of  T. indicum  was significantly influenced by SNP treatments and P stress, with the highest level being obtained when the SNP was 100 μmol/L under low P stress (5 μmol/L). Treatment with 100 μmol/L SNP alone also increased the colonization rate of  T. indicum  and had positive effects on the plant height, stem circumference, biomass, root-shoot ratio and root POD activity of the seedlings at different times after inoculation. Under low P stress, the 100 μmol/L SNP increased the richness of the  norB -type denitrifying bacterial community. Interestingly, the diversity and richness of  norB -type denitrifying bacteria were significantly positively correlated with the colonization rate of  T. indicum . SNP treatments under low P stress altered the abundance of some dominant taxa such as  Alphaproteobacteria ,  Gammaproteobacteria ,  Pseudomonas ,  Ensifer , and  Sulfitobacter.  Evaluation of the fungal community in the rhizosphere revealed that 100 μmol/L SNP treatment alone had no noticeable effect on their richness and diversity, but it did shape the abundance of some fungi.  Buellia ,  Podospora, Phaeoisaria, Ascotaiwania , and  Lophiostoma  were more abundant following exogenous NO application, while the abundance of  Acremonium, Monographella , and  Penicillium  were decreased. Network analysis indicated that  T. indicum  was positively and negatively correlated with some fungal genera when treated with 100 μmol/L SNP. Overall, these results revealed how exogenous NO and P stress influence the symbiosis of truffles and host plants, and indicate that application of SNP treatments has the potential for ectomycorrhizal synthesis and truffle cultivation.

frontiers in microbiology

Exogenous Nitric Oxide and Phosphorus Stress Affect the Mycorrhization, Plant Growth, and Associated Microbes of Carya illinoinensis Seedlings Colonized by Tuber indicum