Bacillus amyloliquefaciens and application thereof
A technology for dissolving amyloid spores and bacilli, applied in the directions of application, bacteria, fungicides, etc., can solve problems such as lack of predictability, and achieve the effects of good inhibitory effect, simple application and operation, and low production cost.
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Embodiment 1
[0023] Embodiment 1, isolation screening and identification of bacterial strain SY-SF-01
[0024] Bacillus amyloliquefaciens SY-SF-01 was isolated from the soil of rice fields in Wuchang City, Heilongjiang Province.
[0025] Separation method: Put 10g of the collected soil sample into a 250mL Erlenmeyer flask filled with 100mL of distilled water, put it in a shaker and vibrate at 28°C for 2 days to make a sample suspension, take 0.5mL of the sample suspension and add it to a 4.5ml Carry out 10-fold serial dilutions in sterile water centrifuge tubes, each take 0.1mL 10 4 、10 5 、10 6 、10 7 The double dilution was applied to peptone medium (sucrose 30g, peptone 5g, FeSO 4 ·7H 2 O 0.01g, MgSO 4 ·7H 2 O 0.5g, KCl 0.5g, K 2 HPO 4 1g, 20g of agar powder, 1000ml of distilled water, pH=7.0~7.2) on a plate, cultivated in a constant temperature incubator at 28°C for 2 days, according to the morphology. According to the size, color and other characteristics, a total of 180 singl...
Embodiment 2
[0058] Embodiment 2, further plate antibacterial effect of bacterial strain SY-SF-01
[0059] Fermentation medium: 30g sucrose, 5g peptone, FeSO 4 ·7H 2 O 0.01g, MgSO 4 ·7H 2 O0.5g, KCl0.5g, K 2 HPO 4 1g, distilled water 1000ml, pH=7.0-7.2
[0060] Preparation of biocontrol bacterium SY-SF-01 fermentation broth: inoculate the SY-SF-01 pure strain obtained in Example 1 into a 250ml Erlenmeyer flask with a liquid volume of 50ml with a sterilized toothpick, at 28°C, the rotation speed Vibrate and cultivate on a constant temperature shaker at 160r / min for 48 hours to obtain a fermented liquid for future use.
[0061] Inoculate the pathogenic bacteria rice sheath blight, cucumber wilt, red bean anthracnose, tobacco target blight, and cotton wilt on PDA medium until the culture dish is covered, and punch out from the edge of the colony with a puncher with a diameter of 6mm. Bacteria cake, inoculate in the center of the blank PDA medium plate, inoculate 1 μl of fermentation b...
Embodiment 3
[0067] Embodiment 3, the preparation of the fermented broth concentrate of bacterial strain SY-SF-01
[0068] The Bacillus amyloliquefaciens SY-SF-01 obtained in Example 1 is inoculated on the peptone solid medium to activate the bacterial strain, and the bacterial strain after the activation is inoculated to the 250ml Erlenmeyer flask with a sterilized toothpick with a filling volume of 50ml, and the 28 DEG C, rotating speed is 160r / min constant temperature shaker culture 24h, make fermented seed liquid, seed liquid is inserted into expansion fermentation medium (composition is the same as fermentation medium in embodiment 2) by volume ratio 5% inoculation amount , the temperature is controlled at 25-30°C, and the rotation speed is 120-200r / min, cultivated for 24h-72h to obtain a fermentation broth, and the concentration of bacteria detected by the dilution plate method reaches 1×10 8 ~1×10 10 Cfu / mL, centrifuge at 1000r / min, take the centrifuged supernatant, and use non-pol...
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