The Conservation and Study of Macromycetes in the Komarov Botanical Institute Basidiomycetes Culture Collection-Their Taxonomical Diversity and Biotechnological Prospects.

Culture collections (CCs) play an important role in the ex situ conservation of biological material and maintaining species and strains, which can be used for scientific and practical purposes. The Komarov Botanical Institute Basidiomycetes Culture Collection (LE-BIN) preserves a large number of original dikaryon strains of various taxonomical and ecological groups of fungi from different geographical regions. Started in the late 1950s for the investigation of Basidiomycetes' biological activity, today, in Russia, it has become a unique specialized macromycetes collection, preserving 3680 strains from 776 species of fungi. The Collection's development is aimed at ex situ conservation of fungal diversity, with an emphasis on preserving rare and endangered species, ectomycorrhizal fungi, and strains useful for biotechnology and medicine. The main methods applied in the collection for maintaining and working with cultures are described, and the results are presented. Some problems for the isolation and cultivation of species are discussed. The taxonomical structure and variety of the strains in the collection fund are analyzed, and they show that the taxonomical diversity of fungi in the LE-BIN is commensurable with the largest CCs in the world. The achievements from the ex situ conservation of the diversity of macromycetes and the main results from the screening and investigation of the collection's strains demonstrate that a number of strains can be prospective producers of enzymes (oxidoreductases and proteases), lipids, and biologically active compounds (terpenoids, phthalides, etc.) for biotechnology and medicine.

Contribution to Studies on Rare Medicinal Mushroom Sparassis crispa (Agaricomycetes) in Culture and Its Production of Phenol Compounds under Various Cultivation Conditions.

Three Sparassis crispa strains from the Komarov Botanical Institute Basidiomycetes Culture Collection (LE-BIN) were studied on various agar and liquid media for growth and phenol compound production. On agar media, the strains produced crystals of various shapes and sizes that glowed in ultraviolet light and were visible to the naked eye. Using gas chromatography-mass spectrometry, the crystals were identified as sparassol (methyl 2-hydroxy-4-methoxy-6-methylbenzoate). Other phenol compounds (i.e., methyl ester of sparassol and methyl ester of orsellinic acid) were also found in S. crispa mycelium. Phenol compounds were detected both in the mycelium and culture filtrate of all studied strains cultivated on various agar and liquid media, at different acidity and duration, but their composition and ratio varied. The initial pH of cultivated medium did not have a large influence on phenol compound production, unlike culture growth duration, which correlated directly with sparassol concentration in mycelium. Production of the phenol compounds presumably connected with constitutive processes of Sparassis metabolism, but the intensity of their production and accumulation in mycelium depends on the strain, conditions, and cultivation time. Strain LE-BIN 2902 is considered a promising sparassol producer.

Diversity of ESI-MS Based Phosphatidylcholine Profiles in Basidiomycetes.

Phosphatidylcholines (PC) are the main membrane lipid constituents comprising more than 50% of total glycerophospholipids. They coordinate a number of cell functions, particularly cell growth, homeostasis, secretion, recognition and communication. In basidial fungi PC are synthesized via the Kennedy pathway as well as through methylation of phosphatidylethanolamines (PE) and then undergo remodeling in Lands cycle that replaces fatty acids in PC molecules. The molecular profile of PC is determined by the genetic features that are characteristic for every species and depend on the environment. Here we present the results of ESI-MS based analyses of PC profiles of 38 species of basidiomycetes belonging to Agaricales (12), Polyporales (17), Russulales (5), Gleophyllales (2), Cantharellales (1), Auriculariales (1), Phallales (1). Although the variety of PC molecular species of basidiomycetes is rather diverse (20-38 molecular species in every profile), only 1-3 main molecular species represent 70-90% of total PC content. The most abundant of them are C36:4 and C36:3, followed by C34:1, C34:2, C36:5, C36:2. In the majority of basidiomycetes, C36:4 reaches up to 50-70% of total PC molecular species. Based on the results of hierarchical cluster analysis four main types of PC profiles which characterized the studied fungi independently from their taxonomic position, ecology, trophic status, and hyphal differentiation have been revealed. Comparative analyses of studied fungi using PCA method have shown that species of Polyporales differ from those of Agaricales by higher variability of PC profiles.