A kind of Bacillus subtilis and its application in agricultural production
A technology for Bacillus subtilis and peanuts, which is applied to a new type of Bacillus subtilis and its application in agricultural production, can solve the problems of unstable use effect, immature research, and few product types, so as to improve quality and achieve agricultural health. , Environmentally friendly effect
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Embodiment 1
[0020] Example 1 Screening of microorganisms with phosphorus dissolving function
[0021] 1. Soil sample: calcareous soil in the southwestern mountainous area of Qingzhou City, Shandong Province.
[0022] 2. Preparation of soil dilution: Weigh 0.5g soil sample and dissolve it in 4.5ml sterile water to make a 1:10 soil solution, then draw 0.5ml soil solution from it and place it in 4.5ml sterile water to make 1:100 Soil solution, and so on, prepare 1:10 6 -10 7 soil dilution solution.
[0023] Take 0.1ml of soil dilution solution and evenly spread it on the insoluble inorganic phosphorus solid medium (10g glucose, (NH) 4 ) 2 SO 4 0.5g, NaCl 0.3g, KC1 0.3g, MgSO 4 ·7H 2 0 0.3g, FeSO 4 .7H 2 0 0.03g, MnSO 4 .4H 2 0 0.03g, Ca 3 (PO 4 ) 2 5.0g, distilled water 1000ml, pH 7.0-7.5, agar 20g, sterilized at 115°C for 30min), invert in a 30°C incubator for 3 days, observe the colonies growing on the medium, 12 of them have obvious colonies around them. The color changes...
Embodiment 2
[0024] Example 2 Rescreening of Phosphorus Resolving Microorganisms
[0025] The 12 strains of bacteria obtained from the primary screening in Example 1 were respectively inoculated on the insoluble inorganic phosphorus solid medium, and after culturing for 3 days at 30°C, the size of the transparent circle around the colony was observed, and it was found that the three strains with the largest transparent circle were LLH- -2, LLH--3, LLH--10.
[0026] The three strains with the largest transparent circle were inoculated into 50 mL of insoluble inorganic phosphorus liquid medium (glucose 10 g, (NH 4 ) 2 SO 4 0.5g, NaCl 0.3g, KC1 0.3g, MgSO 4 .7H 2 0 0.3g, FeSO 4 .7H 2 0 0.03g, MnSO 4 .4H 2 0 0.03g, Ca 3 (PO 4 ) 2 5.0g, distilled water 1000ml, pH 7.0-7.5, sterilized at 115°C for 30min), 30°C, 200rpm, cultured for 6 days, and at the same time, the liquid phosphate solubilization medium without any bacteria was used as the control group; number of viable bacteria.
...
Embodiment 3
[0034] Example 3 Identification of LLH-3 strain
[0035] 1) Colony morphology of LLH-3 strain:
[0036] The colony of the LLH-3 strain is flat, the surface is rough and opaque, the edge is initially smooth and then becomes rough, the colony is white to dirty white, the spore size is about 0.5-0.9×0.8-1.3 μm, oval or columnar, mesophytic or Near middle school students; Gram positive, catalase positive, V.P positive, starch hydrolysis test positive, indole test positive, glucose, arabinose, xylose and mannitol available, growth temperature range 20-45 °C, pH range 5-10.
[0037] 2) Molecular biological identification of LLH-3 strain:
[0038] The LLH-3 strain obtained by the above screening was identified by the method of molecular biology, its 16s rDNA sequence SEQ ID NO: 1 was measured, and blast comparison was performed in the GenBank nucleic acid database.
[0039] SEQ ID NO: 1 is shown below:
[0040]
[0041] Based on the colony morphology of the LLH-3 strain and th...
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