Lactarius vividus is a delicious and precious wild edible mushroom with great economic value. However, as an ectomycorrhizal fungus, it depends on plant symbiosis for its life cycle, resulting in high diffi-culty in artificial cultivation. In the mycorrhizal symbiotic system, mycorrhizal fungi must obtain carbon sources from host plants to fulfill their life cycle. Ectomycorrhiza and arbuscular mycorrhiza (AM) are the main types of mycorrhizae. In arbuscular mycorrhizal symbiosis, fatty acids are the major carbon sources transferred from host plant to the fungal partner. Herein, the modified Melin-Norkrans (MMN) culture medium was supplemented with myristic acid (MYR) or/and peptone (PEP) as growth-promoting components for L. vividus mycelium growth. Measurement of colony on different modified culture media showed that adding both MYR and PEP to the MMN medium could significantly improve the efficiency of L. vividus growth. Nile red staining of mycelial fatty acids showed that L. vividus mycelia absorbed fatty acids from the culture medium as carbon sources for growth, which suggested that ectomycorrhizal fungi have a nutrient exchange mode similar to that of arbuscular mycorrhizal fungi. The results provide a theoretical basis for further research on mycorrhizal symbiosis and cultivation of L. vividus.