Ectomoycorhizal symbiosis between mushrooms and perennial plants is naturally present in almost all land ecosystems. Ectomycorhizal community perennial plants provide organic nutrients for fungus nutrition, and in return acquires an increased absorption surface of the root system and numerous other benefits such as improved herbal nutrition, increased pest tolerance, heavy metal toxicity, drought and salinity, increased growth and yields. In case of truffles in Croatia the mushroom habitats are increasingly depleted, and an alternative is offered to planting truffles. The paper presents the possibilities of producing seedlings for truffle cultivation. This paper will contribute to the knowledge about the possibilities of producing quality planting material for the successful production of truffles.