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The article is devoted to the IBK Mushroom Culture Collection as an essential basis of preservation of macrofungi genofund in pure culture and fundamental research on biology and biotechnology of edible mushrooms with medicinal properties in Ukraine. The objects of investigation are 568 strains of 28 species of edible mushrooms with medicinal properties from the IBK Mushroom Culture Collection. The aim of the work was the elaboration of verification criterions for the strains of edible macromycetes of IBK Collection for confirmation of taxonomic status of biotechnologically valuable producers. The mycological, microbiological, microscopic, molecular genetic methods were done. Methods of macromycetes isolation in pure culture, the criteria for maintaining viability and identification of different species at the vegetative growth stage are provided. Peculiarities of mycelium growth and formation of fruiting bodies in culture are investigated within special program developed by the Collection staff. This program includes research on cultural-morphological characteristics using scanning electron microscopy, cultivation conditions, physiological and biochemical characteristics of strains that can be used to determine species identity of the culture. A complex of morphological criterions for the correct identification of the taxonomic position of cultures of 28 species edible macromycetes are proposed. It includes a presence, dislocation, and morphology of clamp connections; a type of anamorph; special hyphal structures and other characteristics; the presence of teleomorph stage; the growth rate, color, and morphology of mycelial colony. Using light and scanning elec-Chapter «Biological sciences» tron microscopy, new data were obtained on microstructures of vegetative mycelium of valuable species edible mushrooms with medicinal properties from genus Pleurotus (Fr.) P. Kumm., Agaricus L.: Fr., Auricularia Bull. ex Juss., Coprinus Pers., and Lentinula edodes (Berk.) Singer, Hericium erinaceus (Bull.) Pers., Grifola frondosa (Dicks.: Fr.) Gray, Hypsizygus marmoreus (Peck) H.E. Bigelow, Lepista nuda (Bull.: Fr.) Cooke, etc. A list of some important for biotechnology edible mushroom with medicinal properties are described. The content of the main components of fruit bodies of 11 species edible mushroom with medicinal properties maintained in the IBK Collection is provided. The nutritional value of these mushrooms is comparable to foodstuffs such as corn, soybeans, potato. The medicinal properties and bioactive compounds of 8th genera of macromycetes are described.