Actinidia symbiotic Bacillus pumilus rfkf6 strain and its application
A technology of Bacillus pumilus and red heart kiwifruit, which is applied in the field of microorganisms, can solve the problems of blocked aquatic product exports, microbial ecological disorders, and corruption of cold water fish.
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Embodiment 1
[0025] Obtaining Bacillus pumilus RFKF6 strain with high inhibitory activity against Shewanella
[0026] a. Rinse the clean and fresh red kiwi fruit under running water to remove the dust on the surface, sterilize it with 75% ethanol and 2% sodium hypochlorite for 5 minutes in a sterile environment, and then rinse it repeatedly with sterile water. On the ultra-clean workbench, Peel off the skin with sterile scissors, cut the pulp into small pieces with a side length of 1 cm, and place 5 sections on each complete culture dish; culture at a constant temperature of 27°C.
[0027] b. In a sterile environment, use an inoculation needle to pick up the colonies on the plate and repeatedly streak and culture until pure strains appear.
[0028] c. Detection of antibacterial activity: Shewanella (Shewanella) uses beef extract peptone medium. The bacteria were inoculated in beef extract peptone liquid medium, shaken and cultured for 24 hours as indicator bacteria. Use a pipette gun to ...
Embodiment 2
[0030] RFKF6 preservation effect verification experiment
[0031] a. The RFKF strain was cultured with shaking at 30° C. for 30 h in a liquid medium of complete medium. Take the fermentation broth, centrifuge, collect the bacteria and the fermentation supernatant respectively, and suspend the bacteria in sterile water to a final concentration of 10 3 cfu / mL, the strain fermentation supernatant was filtered with a 0.2 μm filter membrane to obtain a sterile fermentation supernatant.
[0032] b. Spray the bacterial suspension and the sterile fermentation supernatant on the surface of the small yellow croaker (blank control without spraying the sample), respectively, pack them into bags and refrigerate at 4°C. Samples were taken on the 0th, 2nd, and 4th day to measure the total number of bacteria and pH value.
[0033] The results showed that with the increase of storage time, the total number of bacteria in small yellow croakers in each group increased, but the increase in the ...
Embodiment 3
[0035] Verification Experiment of Pectinase Production by Bacillus pumilus RFKF6
[0036] a. Rinse the clean and fresh red kiwi fruit under running water to remove the dust on the surface, sterilize it with 75% ethanol and 2% sodium hypochlorite for 5 minutes in a sterile environment, and then rinse it repeatedly with sterile water. On the ultra-clean workbench, Peel off the skin with sterile scissors, cut the pulp into small pieces with a side length of 1 cm, and place 5 sections on each complete culture dish; culture at a constant temperature of 27°C.
[0037] b. In a sterile environment, use an inoculation needle to pick up the colonies on the plate and repeatedly streak and culture until pure strains appear.
[0038] c. screening pectinase producing bacteria test: all pour the pectin agar medium of 15mL in each plate (the formula of described pectin agar medium is: K 2HPO 4 1g, MgSO 4 0.5g, NaNO 3 3g, FeSO 4 ·7H 2 (0.01g, pectin 2g, agar powder 17g, distilled water t...
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