Bacillus velezensis LJBV19 and application thereof
A technology of LJBV19 and Bacillus, applied in the field of microorganisms, can solve the problems of soil microbial richness and soil pH, quality and yield, and inability to obtain fertilizers, etc., to improve fruit quality, develop agricultural economy, and promote agricultural development.
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[0033] (1) Preparation of test medium
[0034] 1) PDA medium: 200g of potatoes, 20g of glucose, 15g of agar powder, 1000mL of distilled water, sterilized by high pressure steam at 121°C for 20min.
[0035]2) LB liquid medium: tryptone 10g, yeast extract 5g, sodium chloride 10g, distilled water 1000mL, adjust pH to 7.0, sterilize by high pressure steam at 121°C for 20min. For LB solid medium, add 15 g of agar powder per liter of distilled water.
[0036] 3) Citrate medium: NaCl 5.0g, MgSO 4 ·7H 2 O 0.2g, NH 4 h 2 PO 4 1.0g, K 2 HPO 4 1.0g, sodium citrate 2.0g, 0.04% phenol red 10mL, agar 15g, distilled water 1000mL.
[0037] 4) Starch agar medium: beef extract 2.0g, egg white 17.0g, agar 15.0g, soluble starch 2.0g, distilled water 1000mL.
[0038] 5) Gelatin medium: 5.0 g of peptone, 120.0 g of gelatin, 3.0 g of beef extract, and 1000 mL of distilled water.
[0039] (2) Cultivation of test plant pathogenic bacteria strains
[0040] M. oryzae, C. viniferum, C. diplo...
Embodiment 1
[0041] Embodiment 1: Isolation and screening of bacterial strain
[0042] (1) Isolation of strains
[0043] The soil was taken from the rhizosphere of grapes, and the sampling requirement was within 20cm from the surface, because the roots of grapes are mainly distributed in this area. Soak the soil with physiological saline (m:v=1:10), oscillate evenly, and let it stand for 30 minutes. After the soil settles, take the supernatant and warm it at 80°C for 20 minutes. After warming, the supernatant was diluted 1, 10, 100, and 1000 times, spread on LB solid medium, cultured at 37°C for 1 day, and selected milky white single plaque with biofilm to be preserved. The 16S rRNA gene was amplified with 27F / 1492R primers, sequenced, compared, and a single colony belonging to Bacillus was retained. A single colony was inoculated in LB liquid medium, and cultured on a shaker at 37°C and 180 rpm for 24 hours to obtain a bacterial suspension. The bacterial suspension was mixed with 50% s...
Embodiment 2
[0047] Embodiment 2: Identification of bacterial strain LJBV19
[0048] (1) Physiological and biochemical characteristics of strain LJBV19
[0049] Observation of colony morphology: Pick an appropriate amount of lawn that has been cultured for 24 hours and inoculate it in LB liquid medium, and shake it on a shaker at 37°C and 180 rpm for 48 hours. Take 3 μL and drop it on the plate, and mainly record the colony shape, size, edge, surface, bulge shape, transparency, and color.
[0050] Gram staining: Pick an appropriate amount of bacterial lawn cultured for 24 hours and spread it evenly on a clean glass slide with a drop of distilled water in the center, air-dry, fix the smear several times over the flame, and stain with crystal violet for 1 min; After rinsing with running water, add iodine solution dropwise for 1 min; after rinsing with running water, decolorize with 95% alcohol for 30 s; after rinsing with running water, counterstain with safranin for 2-3 min; Gram-positive...
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