Endophytic fungus ws01 of Gastrodia elata and its application in increasing the content of gastrodin in Gastrodia elata
A technology of endophytic fungi and Gastrodia elata, applied in the direction of fungi, microorganism-based methods, microorganisms, etc., to achieve the effect of increasing the content of gastrodin
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Embodiment 1
[0039] Isolation and identification of endophytic fungus strain Ws01 from Gastrodia elata
[0040]In November 2017, obtain Gastrodia elata bulbs from Shenpo Mountain, Suining County, Hunan Province, and carry out surface disinfection: rinse the fresh Gastrodia elata bulbs with running water, then soak the cleaned bulbs with 75% ethanol for 3-5min, rinse with sterile water once, and use sodium hypochlorite ( Available chlorine 1.5%) soak for 1-2min, wash 3 times with sterile water, and dry the surface with sterile absorbent paper for later use. Under sterile conditions, take a tissue about the size of mung bean from the center of the corm of Gastrodia elata that has been sterilized on the surface, put it on a PDA solid medium, and culture it statically for 15 days at a temperature of 22-28°C to obtain an endophytic fungus of Gastrodia elata, namely Gastrodia elata endophyte Ws01 strain used in the present invention.
[0041] The Gastrodia elata endophyte Ws01 of the present in...
Embodiment 2
[0045] Application of endophytic fungus Ws01 in Gastrodia elata to increase gastrodin content in Gastrodia elata
[0046] 1. Select the endophytic fungus Ws01 strain we isolated from Gastrodia elata, which is classified as Phlebiopsis sp. Ws01, and is preserved in Guangdong Microbial Culture Collection Center (GDMCC), and the preservation date is June 2020 11th, deposit number: GDMCC No.61055
[0047] 2. Activation and cultivation of strains in inclined test tubes. The endophytic fungus Phlebiopsis sp. (Phlebiopsis sp.) strain Ws01 of Gastrodia elata was inoculated into the slant test tube culture medium with mycelium blocks, and cultured at 25°C for 7-15 days to obtain slant strains;
[0048] Inclined test tube medium is PDA solid medium, 1000ml of PDA solid medium contains potato 200g, glucose 20g, agar 15g, add water to 1000ml, natural pH.
[0049] 3. The expanded culture is carried out by liquid shaking flasks to obtain seed liquid. Put 50ml of liquid shake flask seed c...
Embodiment 3
[0058] Embodiment 3: It is slightly different from steps 3-6 in Embodiment 2, but the rest are the same. Specifically:
[0059] 1. Select the endophytic fungus Ws01 strain we isolated from Gastrodia elata, which is classified as Phlebiopsis sp. Ws01, and is preserved in Guangdong Microbial Culture Collection Center (GDMCC), and the preservation date is June 2020 11th, deposit number: GDMCC No.61055
[0060] 2. Activation and cultivation of strains in inclined test tubes. The endophytic fungus Phlebiopsis sp. strain Ws01 of Gastrodia elata was inoculated into the slant test tube culture medium with mycelium blocks, and cultured at 25°C for 7 to 15 days to obtain slant strains;
[0061] Inclined test tube medium is PDA solid medium, 1000ml of PDA solid medium contains potato 200g, glucose 20g, agar 15g, add water to 1000ml, natural pH.
[0062] 3. Cultivation of Gastrodia elata bulbs: Lay the section wood (generally 30cm long and 7-8cm wide) on the bottom of the cellar bed (t...
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