The utilization of fast-growing, economically valuable woody plants with strong stress resistance, such as poplar and willow, to revegetate severely metal-contaminated mine tailings not only offers a productive and profitable use of abandoned polluted soil resources but also facilitates the phytoremediation of these polluted soils. This study examines the diversity and functional roles of endophytic fungi naturally colonizing the roots of an artificially established Populus yunnanensis forest and the naturally reclaimed pioneer species Coriaria sinica on an abandoned tailing dam in southwest China. Culture-independent analyses revealed that the root systems of both plant species were abundantly colonized by arbuscular mycorrhizal fungi and endophytic fungi, forming rich and diverse endophytic fungal communities predominantly represented by the genera Ilyonectria, Tetracladium, Auricularia, and unclassified members of Helotiales. However, the composition of root endophytic fungal communities differed significantly between the two plant species. Using a culture-dependent approach, a total of 192 culturable endophytic fungal strains were isolated from the roots. The dominant genera included Cadophora, Cladosporium, Cyphellophora, and Paraphoma, most of which were previously identified as dark septate endophytes (DSE). Six representative DSE strains were selected for further study, and significant cadmium tolerance and various plant growth-promoting traits were observed, including the solubilization of insoluble inorganic and organic phosphorus, indole-3-acetic acid (IAA) production, and siderophore synthesis. In greenhouse experiments, inoculating two DSE strains mitigated the inhibitory effects of metal-polluted tailing soil on the growth of P. yunnanensis. This was achieved by reducing heavy metal uptake in roots and limiting metal translocation to the above-ground tissues, thereby promoting plant growth and adaptability. Our findings suggest that as plants reclaim metal-polluted tailings, root-associated endophytic fungal communities also undergo natural succession, playing a critical role in enhancing the host plant’s tolerance to stress. Therefore, these restored root-associated fungi, particularly DSE, are essential functional components of the root systems in plants used for tailing reclamation.
