Selection and evaluation of conservation conditions of Claviceps purpurea saprophytic lines

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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.

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About the authors

A. A. Volnin

All-Russian Scientific Research Institute of Medicinal and Aromatic Plants

Author for correspondence.
Email: volnin.a@mail.ru
ORCID iD: 0000-0001-9222-536X

Ph.D. (Biol.), Leading Research Scientist, Laboratory of Biotechnology

Russian Federation, 7, ul. Grina, Moscow, 117216

R. I. Bobyleva

Email: volnin.a@mail.ru

к.б.н.

Russian Federation

N. S. Tsybulko

All-Russian Scientific Research Institute of Medicinal and Aromatic Plants

Email: volnin.a@mail.ru
ORCID iD: 0000-0001-9542-5017

Ph.D. (Pharm.), Engineer, Laboratory of Biotechnology

Russian Federation, 7, ul. Grina, Moscow, 117216

P. S. Savin

All-Russian Scientific Research Institute of Medicinal and Aromatic Plants

Email: volnin.a@mail.ru
ORCID iD: 0000-0002-5441-3471

Ph.D. (Biol.), Leading Research Scientist, Laboratory of Biotechnology

Russian Federation, 7, ul. Grina, Moscow, 117216

K. A. Shtrom

MIREA – Russian Technological University (Moscow State University of Fine Chemical Technologies named after Lomonosov)

Email: volnin.a@mail.ru

Master

Russian Federation, Vernadsky Avenue, 86, Moscow, 119454

S. B. Myasnikova

All-Russian Scientific Research Institute of Medicinal and Aromatic Plants

Email: volnin.a@mail.ru
ORCID iD: 0000-0002-6318-2433

Research Scientist, Laboratory of Biotechnology

Russian Federation, 7, ul. Grina, Moscow, 117216

A. I. Bokhan

All-Russian Scientific Research Institute of Medicinal and Aromatic Plants

Email: volnin.a@mail.ru
ORCID iD: 0000-0003-4154-3709

Dr.Sc. (Agrical.), Head of  Laboratory of Biotechnology

Russian Federation, 7, ul. Grina, Moscow, 117216

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Supplementary files

Supplementary Files
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1. JATS XML
2. Figure. Mycelium of super-elite lines: A –presence of ergoalkaloids in two mycelium samples from new ergotoxin lines on 30th day of cultivation; Б – pigmented purple mycelium, capable of synthesizing alkaloids in vitro; absence of typical hyphal morphology indicates to formation of an atypical sclerotia-like mycelium; B – non-pigmented (white) mycelium, unable to synthesize alkaloids during in vitro cultivation; typical characteristic hyphal morphology of Ascomycota

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