A kind of bacillus amyloliquefaciens jspb14 and its application
A technology for dissolving Bacillus amyloliquefaciens and bacilli, applied in the fields of application, bacteria, chemicals for biological control, etc. Reduced residual, high antagonistic activity
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Embodiment 1
[0048] Isolation and identification of Bacillus amyloliquefaciens strain JSPB14
[0049] (1) Separation of strain resources to be screened: randomly sample 50 g from the cultivation substrate with a higher incidence of postharvest Pleurotus eryngii canker, take 5 substrate samples in total, mix well, take 10 g of substrate and put it into 20 grains of fine Glass beads and 100 mL of sterilized deionized water were placed in a 250 mL Erlenmeyer flask, placed on a shaker at 150 r / min, and shaken at 30 ° C for 6 h. After standing still for 10 minutes, take 1 mL of the supernatant, dilute it 10-fold with sterilized deionized water, draw 100 μL of each concentration gradient dilution, and spread it on a 2YT solid plate, and culture it statically at 30°C for 36 hours. Use an inoculation loop to pick a single colony and streak and purify it twice on 2YT solid medium, transfer the purified single colony with different forms to a 2YT solid plate, culture it statically at 30°C for 36h, a...
Embodiment 2
[0054] Growth Dynamics of JSPB14 in Landy's Medium and Antibacterial Activity of Secondary Metabolites
[0055] Take out the JSPB14 strain preserved in glycerol from the -70°C ultra-low temperature refrigerator, inoculate it on a 2YT solid plate, and culture it at 30°C for 36 hours; use a sterilized toothpick to pick a single colony of JSPB14 and inoculate it into a 250mL conical tube containing 50mL 2YT liquid medium In the flask, cultured at 150r / min and 30°C for 36h as the seed solution; inoculated the seed solution into a 250mL Erlenmeyer flask containing 50mL Landy liquid medium at a ratio of 5% (v / v), 75r / min, Cultivate for 60 h at 30°C. Investigate the bacterial population density at 0h, 4h, 8h, 12h, 18h, 24h, 36h, 48h, and 60h after inoculation. The specific method: take samples at the above time points, dilute 10 times with sterile water, and draw 100 μL of each A 2YT solid plate was coated with a concentration gradient dilution solution, and cultured at 30°C for 48 ...
Embodiment 3
[0058] Determination of Antibacterial Spectrum of Biocontrol Strain JSPB14
[0059] In order to evaluate the antibacterial spectrum of the secondary antibacterial substances of Bacillus amyloliquefaciens strain JSPB14, six major crop pathogenic bacteria and one human opportunistic pathogenic bacteria were selected for plate antagonism experiments, including Lactococcus lactis subsp. subsp.lactis), Xanthomonas oryzaepv.oryzae, Xanthomonas oryzae pv.oryzicola, Xanthomonas campestris pv.camestris, Erwinia carotovora subsp. carotovora, Ralstonia solanacearum and Staphylococcus aureus. Specific method: Take out the above-mentioned 6 kinds of glycerol strains of pathogenic bacteria except Lactococcus lactis subspecies lactic acid from the ultra-low temperature refrigerator at -70°C, respectively streak on 2YT solid medium, culture at 30°C for 36 hours, and pick single colonies respectively In 2YT liquid medium, cultivate to OD at 30°C and 150r / min 600 ≈0.5, for later use; take out...
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