In this study, we investigated the culture conditions of four ectomycorrhizal fungi, namely, Amanita spissacea NIFoS 2719, Pisolithus arhizus NIFoS 2784, Suillus spraguei NIFoS 2848, and Xanthoconium affine NIFoS 2716, in solid and liquid culture media. In addition, we attempted to induce in vitro mycorrhization of the fungi with Pinus densiflora. Prior to liquid culture, we determined the optimal culture conditions for each species in solid media. The results revealed that all species examined are capable of growth in potato dextrose agar (PDA), malt extract agar (MEA), and modified Melin-Norkran’s medium (MMN), although their preferred growth media were different. Liquid culture experiments showed that inorganic nitrogen did not enhance the mycelial growth of all four species. Therefore, we used MMN-based liquid inocula to promote the growth of ectomycorrhizal fungi in our symbiosis culture system. Mycorrhization was observed in Xanthoconium affine NIFoS 2716. Morphological analysis revealed that fungi-inoculated roots of P. densiflora form simple and dichotomous lateral roots with dense mycelia. In addition, inoculation with X. affine NIFoS 2716 promoted root and shoot developments.