Arbuscular mycorrhizal fungi (AMFs) and biochar are two common alternatives to chemical fertilizers applied to soil to improve crop growth. However, their interactive effects on maize (Zea mays L.) growth, nutrient absorption, and physiological properties remain poorly understood. In this study, maize plants were grown in pots treated with biochar and AMFs Diversispora eburnea, alone or in combination. The results showed that the individual application of AMFs or biochar increased maize growth and mineral contents in shoots and roots (including P, K, Ca, Na, Mg, Fe, Mn, and Zn). The chlorophyll a, chlorophyll b, and total chlorophyll contents in AMF-treated leaves were significantly higher than those in the control treatment group. However, AMFs had no synergistic effects with biochar on maize growth, nutrient absorption, nor photosynthetic pigments. The application of biochar to the soil significantly reduced mycorrhizal colonization by 40.58% in the root tissues, accompanied by a significant decline in mycorrhizal dependency from 80.57% to -28.67%. We conclude that the application of biochar and AMFs can affect maize growth, nutrient uptake, and physiological properties. Our study can provide vital information for further resource use optimization in agroecosystems.