Separation and application of antiviral bacillus amyloliquefaciens
A technology of amylolytic spores and bacilli, applied in the direction of bacteria, microorganism-based methods, microorganisms, etc.
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Embodiment 1
[0026] Embodiment 1, the acquisition of Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) YHN
[0027] The strain (YHN) was isolated from the rhizosphere soil of edamame in Heze. Take 10 g of soil samples from the rhizosphere of edamame and add them into a triangular flask filled with 100 mL of sterile water to make a soil suspension. Put the Erlenmeyer flask on a shaker, shake it for 20 minutes, and take the supernatant after standing for 5 minutes. Using the 10-fold serial dilution method, draw 200 μL of each concentration on the broth peptone agar medium (beef extract 3g, peptone 10g, NaCl 5g, agar 20g, distilled water supplemented to 1000mL, pH 7.2-7.4) and spread evenly on the plate. concentration repeated three times. Then the broth-peptone agar medium plate was cultured upside down in a biochemical incubator at 28°C for 24 hours. When colonies appear, spray the indicator bacterium Geotrichum candidum (Geotrichum candidum) spore suspension, and it is advisable t...
Embodiment 2
[0070] Embodiment 2: the mensuration of YHN antibacterial spectrum
[0071] The antibacterial effect of the 8 pathogenic bacteria listed in Table 1 was determined by confrontation culture method (Sijam et al., 2005). The above-mentioned pathogenic bacteria cakes with a diameter of 9 mm were respectively connected to the center of a potato dextrose agar plate with a diameter of 90 mm, then the plate was sealed with a parafilm, and put into a biochemical incubator at 28° C. for two days. Dip the sterilized filter paper with a diameter of 6mm into the prepared Bacillus amyloliquefaciens YHN bacterial solution (107cfu / mL), move it to a distance of 15mm from the edge of the pathogenic bacteria, seal it, and finally put it in a biochemical incubator at 28°C for cultivation ( figure 1 ). Each pathogen in Table 1 was treated according to the above steps and repeated 3 times (repeated). After 7 days, the diameter of the inhibition zone was measured, and the inhibition rate was calcul...
Embodiment 3
[0074] Example 3: Indoor inhibitory effect of YHN on tobacco mosaic virus
[0075] The YHN was streaked on the broth peptone agar medium plate, and the plate was placed in a biochemical incubator at 28°C for 12 hours. Pick a single colony with a toothpick and put it into a test tube filled with 5 mL of liquid LB medium (tryptone 10 g, yeast extract 5 g, NaCl 10 g, add water to volume 1 L, autoclave), and shake at 28 ° C for 14-16 h.
[0076] With the inoculum amount of 1:100, the above-mentioned culture solution was inoculated into a 100 mL Erlenmeyer flask, and cultured with shaking at 28° C. for 14-16 h. Centrifuge the bacterial solution to obtain the bacterial cells, rinse with an appropriate amount of sterile water, resuspend, centrifuge again, and repeat the washing twice; obtain the bacterial cells without LB medium, and finally adjust the bacterial cells to a final concentration of 10 with sterile water. 7 cfu / mL.
[0077] Select Sansheng tobacco (a variety of common ...
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