Selection of Pure Strain of Ophiocordyceps sinensisfrom Dried Fruiting Bodies Sourced from China
DOI:
https://doi.org/10.14456/jare-mju.2024.48Keywords:
Strain selection, Identification, Morphology, Ophiocordyceps sinensisAbstract
Ophiocordyceps sinensis is a medicinal mushroom that has been used in traditional Chinese medicine for a long time. Its main bioactive constituents are polysaccharides, cordycepin, and adenosine, which contribute to body nourishment and outstanding pharmacological effects. That is, it can fight cancer and stimulate the immune system by reducing antioxidants and hypoglycemia. The aforementioned mushrooms are in demand and have a high market value, causing a crisis of insufficient natural raw materials. The cultivation of mycelium to replace mushrooms was used to supply market demand.
This study was conducted to obtain the correct pure strain of the O. sinensis fungus. Fungal morphological, nucleotide sequencing, and phylogenetic analyses were used. Naturally dried O. sinensis mushrooms were collected from various sources in China by using both the part of the fungus growing from the worm's head (Stroma) and the part of the worm's body (Sclerotium). The results of this research showed that one isolate had colony characteristics and morphology similar to the O. sinensis fungus. Using molecular biology techniques by comparing the nucleotide sequence of nuclear ribosomal RNA gene amplified with primers ITS4 (5′-TCCTCCGCTTATTTGATATGC-3′) and ITS5 (5′-GGAAGTAAAAGTCGTAACAAGG-3′), it was confirmed that the fungus was O. sinensis. When comparing the phylogeny of the phylogenetic tree, it was found that O. sinensis was in the family Ophiocordycipitaceae.
Cultivation of pure strains of fungus could be cultured in an agar formula of 10 g of peptone, 40 g of dextrose, 20 g of yeast extract, 15 g of agar powder, and 1 L of distilled water at a temperature of 16˚C in the dark for 14 days. Morphological characteristics of the pure fungus that was found could be explained as a round colony with white to light cream at the center with white hyphae and densely stacked feathers, branches, fluffy aerial mycelium, and rod-shaped spores of 5-10 µm in size. As a result, the pure fungal strain that is produced can be utilized as an inoculum for the identification of biological components and as material for additional research on their pharmacological effects.
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