SELECTION AND EVALUATION OF CONSERVATION CONDITIONS OF CLAVICEPS PURPUREA SAPROPHYTIC LINES

Abstract

Introduction. Ergot Claviceps purperea plays an important role in health-care technologies. Accumulation of genetic resources of industrial pharmaceutical strains of parasitic ergot is the actual scientific problem. This work is devoted to development of selection and evaluation of conservation conditions of Claviceps purpurea saprophytic lines. Conidiospores and saprophytic mycelium were frozen at –20 0C, after 10 months of storage and cultivated in Erlenmayer flasks on a liquid medium and on an agarised medium. Cryopreserved samples of conidiospores and mycelium retained the ability to grow and alkaloids biosynthesis in vitro. Results. The best lines were selected from sclerotias of 3 parasitic strains and deposited in the form of axenic and saprophytic mycelium for long-term storage (ergotoxin ВКМ-F-2450-D-23-1, ergotoxin ВКМ-F-2450-D-23-9, ergotamine ВКМ- F-2641-D-S-2, ergocornam ВКМ-F-3662-D-22-5). Sclerotia of ergotamine-producing strain had a level of alkaloid synthesis of 0.78 g/100 g, with an ergotamine content of 67.9%, and demonstrated of pigmented purple mycelium in axenic culture. Sclerotias of ergotoxin-producing strain – 0.76 and 0.59 g/100 g, containing 51.5 and 50.3% of the sum of ergocornine and α-ergocryptine, 20 and 22% of β-ergocryptine, respectively, both lines demonstrated sclerotia-like mycelium and alkaloids biosynthesis in saprophytic cultivation on agarised medium. Sclerotia of ergocornam-producing strain – 0.79 g/100 g, containing 18.2% of ergocornam and 24.6% of ergometrine, demonstrated pigmented purple mycelium in axenic culture. Conclusion. Development of long-term storage biocollections, including cryocollections for organisms producing pharmaceuticals raw substances, creates the basis for technological sovereignty of the Russian Federation. It is reducing dependence on imported drugs and pharmaceuticals raw substances, including ergoalkaloids.