Atractylodes macrocephala endophytic fungus, fermentation product and application thereof
A technology of endophytic fungi and fermentation products, applied in the direction of application, microorganisms, fungi, etc., to achieve the effect of inhibiting growth and enhancing resistance
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Embodiment example 1
[0025] The endophytic fungus is isolated and obtained according to the following steps: quickly rinse the soil and impurities on the surface of Atractylodes macrocephala tuber with clean water, after rinsing, filter paper to dry the surface moisture, follow the steps of 75% ethanol for 1 min, 2.5% sodium hypochlorite for 5 min, and 75% ethanol for 1 min After surface disinfection, dry the water with sterilized filter paper, then cut the root tubers into small tissue pieces (0.5 cm×0.5 cm) with a sterilized scalpel, randomly pick tissue pieces, and arrange them in groups of 4 On a plate of PDA medium containing penicillin (50mg / L). After sealing, culture at 26°C, regularly observe the growth of endophytic fungal hyphae and the formation of colonies, and cultivate to a single colony after continuous separation and purification. The endophytic fungi of a single colony were divided into different morphotypes according to the morphology of the hyphae and the culture characteristics...
Embodiment 2
[0028] Take the cryopreservation tube of the AM569 bacterial strain, under aseptic conditions, pick a small amount of mycelia with an inoculation needle, and insert it into the sterilized potato glucose medium (the formula is 200g potatoes, 20g glucose, 15g agar, 1000mL distilled water, natural pH ) at 26°C and 65% humidity for 7 days in dark; under aseptic conditions, use a 5mm hole puncher to punch out the bacterial blocks at the edge of the colonies of the above strains, transfer them to 50mL PDB medium, and incubate at 26°C, Under the condition of constant temperature of 180r / min, the shaking culture was shaken for 8 days; the fermentation product was filtered by suction to obtain the fermentation filtrate and mycelium, and the mycelium was dried (50°C) and weighed, and the mycelium was crushed to make a powder. Methanol (mL): Mycelia (g) = 20:1 ratio of ultrasonic extraction for 1 hour, and suction filtration to obtain the mycelium extract; the fermentation filtrate was ex...
Embodiment 3
[0030] Get Atractylodes macrocephala root rot pathogenic bacteria Fusarium oxysporum and semi-naked Fusarium cryopreservation tubes, under aseptic conditions, pick a small amount of hyphae with an inoculation needle, insert the sterilized potato glucose medium (the formula is 200g of potatoes, glucose 20g, 15g of agar, 1000mL of distilled water, natural pH), 26°C, 65% humidity and dark culture for 5d; In the PDB medium, shake the shaker at 26°C and 180r / min at a constant temperature for 5 days, filter it with 4 layers of sterile gauze, take the filtrate, and mix the mixed bacterial suspension of Fusarium oxysporum and Fusarium seminude in equal proportions , and adjusted to OD600 = 0.6 ~ 1.0, that is, the bacterial suspension of pathogenic bacteria. Take Atractylodes macrocephala seedlings that have grown to 5cm in height, cut 2-3cm wounds on the main root with a sterile knife, soak in the above-mentioned pathogenic bacteria mixed suspension for 10min, and culture at 26°C for ...
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