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[Objective] By examining intracellular and extracellular metabolite changes in ectomycorrhizal fungi (ECMF) under acidic aluminum stress, we identified key resistance-related metabolites and pathways, aiming to elucidate the aluminum tolerance mechanisms from the perspective of metabolic physiology and offer a theoretical basis for using ECMF in restoring aluminum-contaminated forests. [Methods] Pisolithus tinctorius was cultured in vitro in the acidic medium (pH 3.8) containing 0.0 mmol/L or 1.0 mmol/L Al3+. Untargeted metabolomics was employed to analyze changes in intracellular and extracellular metabolite levels. [Results] Compared with that under the 0.0 mmol/L Al3+ treatment, the colony diameter of P. tinctorius under 1.0 mmol/L Al3+ stress decreased significantly by 23.67%. In addition, the intracellular levels of nucleotides including uridylic acid, cytidine monophosphate, uridine, uridine diphosphate, cytidine, and guanosine were upregulated under 1.0 mmol/L Al3+ stress. Extracellular levels of organic acids such as shikimic acid, fumaric acid, heptanoic acid, and tartaric acid, along with carbohydrates including l-arabinose, trehalose, sucrose, and glucose, were also upregulated. Pyrimidine metabolism and citric acid cycle pathways were enriched intracellularly, while ABC transporters and phosphotransferase system pathways were enriched extracellularly. The potential biomarkers identified in the intracellular environment was citric acid, and those identified in the extracellular environment were trehalose and tartaric acid. [Conclusion] Acidic aluminum stress inhibits the growth of P. tinctorius. Intracellularly, P. tinctorius maintains cellular homeostasis and energy supply through enhanced nucleotide accumulation and activation of the citric acid cycle. Extracellularly, P. tinctorius promotes organic acid secretion and carbohydrate efflux to resist aluminum toxicity and associated oxidative damage.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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