Bacillus amyloliquefaciens and application thereof
A technology of amylolytic spores and bacilli, which is applied in the direction of application, bacteria, and fungicides, and can solve problems such as unstable application effects and single function of bacteria species
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Embodiment 1
[0119] Example 1. Isolation, Identification and Preservation of Bacillus amyloliquefaciens FH1 CGMCC No.17050
[0120] 1. Separation
[0121]1. Add 10g soil sample (collected from the rhizosphere soil of tea trees in Xixinan Town, Huizhou District, Huangshan City, Anhui Province) in 90mL sterile water, vibrate at high speed for 10min, and let stand for 30s to obtain the supernatant; put the supernatant in Heat in boiling water at 100°C for 10 minutes, cool naturally to obtain a soil suspension (at this time, the dilution is recorded as 10 -1 ).
[0122] 2. Take 1mL of soil suspension and add it into a test tube filled with 9mL sterile water and mix well (the dilution is 10 -2 ), then draw 1mL from this test tube and add it to another test tube filled with 9mL sterile water and mix well to make a dilution (the dilution is 10 -3 ).
[0123] 3. Spread the diluted solution evenly on the LB solid medium, and incubate at 30°C for 3 days. Purify the bacterial colonies that can g...
Embodiment 2
[0143] Embodiment 2, the mensuration of Bacillus amyloliquefaciens FH1 nitrogen-fixing ability
[0144] Ashbey solid medium: add mannitol 10.0g, KH to appropriate amount of distilled water 2 PO 4 0.2g, MgSO 4 ·7H 2 O0.2g, NaCl0.2g, CaCO 3 5.0g, CaSO 4 2H 2 O0.1g and 20g of agar powder, adjust the pH value to 7.0-7.5, then dilute to 1.0L with distilled water, and sterilize at 121°C for 15min.
[0145] 1. A single clone of Bacillus amyloliquefaciens FH1 was inoculated into 10 mL of LB liquid medium, and shaken at 30° C. and 180 rpm for 48 hours to obtain a culture solution.
[0146] 2. After completing step 1, take the cultured bacteria liquid, centrifuge at 4°C and 10,000 rpm for 5 minutes, and collect the bacteria.
[0147] 3. After completing step 2, take the bacteria and dilute with sterile water to obtain a concentration of 1×10 8 Individual / mL of Bacillus amyloliquefaciens FH1 bacterial suspension.
[0148] 4. After completing step 3, spot 5 μL of the bacterial s...
Embodiment 3
[0150] Embodiment 3, the determination of Bacillus amyloliquefaciens FH1 potassium decomposing ability
[0151] Potassium solution solid medium: Add 2.0 g of disodium hydrogen phosphate, 0.5 g of magnesium sulfate heptahydrate, 0.005 g of ferric chloride, 0.1 g of calcium carbonate, 5.0 g of sucrose, 1.0 g of potassium feldspar powder and 20 g of agar powder into appropriate amount of distilled water, Adjust the pH value to 7.0-7.5, then dilute to 1.0L with distilled water, and sterilize at 121°C for 15 minutes.
[0152] Spot 5 μL of the bacterial suspension of Bacillus amyloliquefaciens FH1 prepared in step 3 of Example 2 on the potassium-dissolving solid medium, culture at 30°C for 3 days, and then make the following judgment: if Bacillus amyloliquefaciens FH1 can be cultured on the potassium-dissolving solid Bacillus amyloliquefaciens FH1 has the ability to decompose potassium; if Bacillus amyloliquefaciens FH1 cannot grow on the solid medium for decomposing potassium, then...
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