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Niche construction by biota has been frequently reported for animals and plants, but fungi have received less attention. For mycorrhizal fungi, mutualistic niche construction has been proposed where partners construct niches for each other. However, few eco-evolutionary mechanisms leading to mutualistic symbiotic feedback have been described. Here we report niche construction by two ectomycorrhiza-forming species of truffles, Tuber aestivum and T. melanosporum. The soils of 263 spots of these Tuber, which create bare patches (brûlés), have been monitored (9–10 years) in natural forests to analyze whether they meet the criteria to satisfy the mutualistic niche construction process. Soil habitat modification by these Tuber can be seen in their brûlés, where the vegetation of arbuscular mycorrhizal plants and soil organisms are largely suppressed. Tuber melanosporum brûlés were smaller more productive, and …