Paenibacillus polymyxa for preventing and treating gray mold of paris polyphylla and application of paenibacillus polymyxa
A technology for polymyx spore and botrytis cinerea, which is applied in the field of polymyxa spore for preventing and controlling botrytis cinerea, can solve problems such as no medicine available, excessive pesticide residues, environmental pollution, etc., so as to increase microbial diversity, Increased income, good compatibility
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Embodiment 1
[0036] The isolation, screening and identification of embodiment 1 target bacterial strain
[0037] The target strain was isolated from the stems of Chonglou in Enshi City, Hubei Province, and healthy stems of Chonglou were taken, surface-sterilized with 75% alcohol for 1 minute, rinsed with sterile water for 3 times, air-dried under sterile conditions, removed the epidermis, and mixed with sterile water Grind, absorb the mixed solution and dilute it, spread it on a PDA plate, incubate at 28°C for 72 hours, and pick a single colony for purification and culture. Botrytis cinerea cinerea 3cm, according to the criss-cross method, place the isolated endophytic bacteria of the stem of Prunus cinerea. After 3 days, when the control plate is full, measure the distance from the outer edge of the bacterial colony to the hyphae of gray mold, repeat 3 times, and select The bacteria with the largest distance were used for later research.
[0038] The target strain is Gram-positive bacter...
Embodiment 2
[0041] The preparation of embodiment 2 bacterial agents
[0042] S1. The endophytic bacterium Paenibacillus polymyxa was taken with an inoculation loop and spread on the PDA medium, and cultured at 28° C. for 72 hours. The formula of the PDA solid medium is as follows: take 200 grams of potatoes, 20 grams of glucose, 15-20 grams of agar, 1000 milliliters of distilled water, pH 7.2-7.4, and sterilize under high pressure steam at 121 ° C for 30 minutes.
[0043] S2. Transfer the single colony activated in step S1 into a 250ml Erlenmeyer flask with 100ml of PDB liquid medium, and culture at 200rmp and 28°C for 72 hours to obtain a fermented seed solution. The formula of PDB liquid medium is as follows: take 200 grams of potatoes, 20 grams of glucose, 1000 milliliters of distilled water, pH 7.2-7.4, and sterilize at 121 ° C for 30 minutes with high-pressure steam to obtain the product.
[0044] S3. Inoculate the inoculation amount of the fermented seed liquid in step S2 into the ...
Embodiment 3
[0050] The influence of embodiment 3 target bacterial strain fermentation supernatants on the cinerea cinerea
[0051] Take 500 μL of the activated target bacterial solution and inject it into 100 mL of PDB, and incubate at 28°C and 180 r / min for 72 hours to obtain the fermentation stock solution. The fermentation stock solution was centrifuged at 12,000 r / min for 10 min, the supernatant was collected, and the fermentation filtrate was obtained by filtering with a 0.22 μm microporous membrane. Get filtrate and the PDA mixing of about 45 ℃, make the final concentration of filtrate be 5% and 10%, inoculate Botrytis cinerea mycelia block (diameter 0.5cm) in plate center after cooling, set blank control, each handles three repetitions, Cultured in a 21°C incubator. Measure the diameter of the colony after the control plate is overgrown (4 days), and calculate the inhibition rate. Inhibition rate (%) = (colony diameter of pathogenic bacteria in the control group - colony diameter...
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