Soil chromium (Cr) contamination has become an environmental problem of global concern. However, the joint effects of combined utilization of biochar and arbuscular mycorrhizal (AM) fungal inoculum, which are considered as two promising remediation strategies of soil heavy metal pollutions, on plant Cr resistance are still poorly understood. In this study, a two-factor pot experiment was conducted to investigate how biochar and AM fungus Rhizophagus irregularis regulate Medicago sativa growth, physiological trait, nutrient and Cr uptake, relevant gene expressions, soil properties, and Cr speciation, independently or synergistically. The results showed that biochar notably decreased AM colonization, while biochar and AM fungus could simultaneously increase plant dry biomass. The greatest growth promotion was observed in mycorrhizal shoots at the highest biochar level (50 g kg−1 soil) by 91 times. Both biochar application and AM fungal inoculation enhanced plant photosynthesis and P nutrition, but the promoting effects of AM fungus on them were significantly greater than that of biochar. In addition, the combined application of biochar and AM fungus dramatically reduced shoot and root Cr concentrations by up to 92 % and 78 %, respectively, compared to the non-amended treatment. Meanwhile, down-regulated expressions were observed for metal chelating-related genes. Furthermore, Cr translocation from roots to shoots was reduced by both two soil amendments. Transcriptional levels of genes involved in reactive oxygen species and proline metabolisms were also regulated by biochar application and AM fungal colonization, leading to alleviation of Cr phytotoxicity. Furthermore, AM fungal inoculation slightly elevated soil pH but decreased plant-available soil P, which was, by contrast, lifted by biochar addition. The combined application reduced soil acid-extractable Cr concentration by 40 %. This study provides new insights into comprehensively understanding of the mechanisms of biochar and AM fungi combination on improving plant Cr tolerance.
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