OPTIMIZATION OF NUTRITION MEDIA AND CONDITIONS FOR IN VITRO CULTIVATION OF EVERNIA PRUNASTRI (L.) ACH.

The work investigated the optimization of nutrient media and in vitro cultivation conditions of the lichen E. prunastri. A study of the growth and development of the culture under controlled conditions revealed a number of patterns at the morphometric and biochemical level. To select optimal conditions for in vitro cultivation of natural isolates, it was adopted to use various variants of the Murashige-Skoog nutrient medium with the addition of alcohol extracts or aqueous decoctions of bark from plants on which the lichen grows naturally (oak, apple, sloe, pear). The growth parameters of E. prunastri isolates have been established. The influence of cultivation conditions on the development of lichen in vitro and the production of biologically active pharmacologically valuable substances was studied. The viability and level of biomass accumulation of E. prunastri isolates from various natural habitats on Hoagland’s medium varied significantly (3–13 times), which indicates the need to select a highly active producer. The size of lichen cells when grown in vitro in a liquid nutrient medium is higher than in natural conditions (by 20.0–38.5 %%). It has been shown that the best medium for the accumulation of biomass and biologically active compounds (lichen acids, chlorophyll) is the Murashige-Skoog variant with the addition of apple bark decoction when grown in diffused light (1000–1500 lux) with a 16-hour photoperiod, temperature 18+1oC and humidity 90+5 %. The chemical analysis of the raw materials of cultivated and wild-growing lichen, as well as the resinoid (thick extract) based on it, indicates a similar qualitative and quantitative composition, which indicates the prospects of the biotechnology for their production. A comparative assessment of natural isolates of E. prunastri and determination of the possibilities of their cultivation in vitro in connection with the development of innovative lichen technology significantly expands the understanding of the biotechnological aspects of obtaining the “oak moss” resinoid (thick Evernia extract) for use in the perfumery, cosmetics, food and pharmaceutical industries as an odor fixer, aromatic and pigment flavoring agent, a source of biologically active pharmacologically effective compounds.