Tricholoma matsutake and Rhizopogon roseolus form ectomycorrhizal (ECM) association with their host plant on natural habitats. The main objective of this study was to test mycelial growth, morphology, and host plant survival both in vitro and in vivo when treated with enriched media. Aseptically germinated seedlings of Pinus densiflora and P. thunbergii were inoculated with the strains of T. matsutake and R. roseolus, respectively. Under in vitro conditions mycelial growth rates performed best on pH 5 and were better on Modified-Melin-Norkrans-(MMN) based medium and Potato Dextrose Agar (PDA); addition of micronutrients and vitamins in MMN mycelial growth rates had 6-27% differences. Without ECM, plant survival rates on standard media were 30% to below 30% and by inclusion of elements they were 50% to 80%. On in vivo, soil containing different media with ECM allowed successful mycorrhizal association and increased seedling survival rates approximately 100%. Our findings confirm that MMN and PDA allowed higher mycelial growth but poor plant survival (<30%); however, enriched media supported 100% plant survival with successful ECM associations. The present method is advantageous in terms of giving objectivity for ECM by employing suitable media for strains and host plant, and making it possible for mass production of ECM-infected seedlings.