A kind of bacillus cereus bacterial strain and application
A technology of Bacillus cereus, which is used in the field of microorganisms, can solve problems such as damage and microbial infection, limit the development of the strawberry industry, and affect the interests of producers, and achieve strong tolerance, easy cultivation and preservation, and reliable sources. Effect
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Embodiment 1
[0014] Example 1: Isolation of Bacillus cereus zy-1 of the present invention
[0015] Isolation and purification of antagonistic bacteria: ①Weigh 10g wheat field soil sample taken from Zaoyang City, Hubei Province, ②Place it in 200mL sodium hexametaphosphate solution (dispersant), shake and suspend for 5min, ③In a suspended state, absorb Add 20mL to 80mL of 0.1% water agar, shake and suspend for 5 minutes, ④When the suspension is in a stirring state, absorb 0.3mL of the suspension, ⑤Dilute the 0.3mL suspension by 10 times, respectively -1 , 10 -2 And 10 -3 , ⑥ Place the gradient dilutions in an oven at 80°C for 30 min, take them out and wait to cool down, ⑦ spread 3 plates (3 plates for each dilution concentration), 1 / 10 TSA selective medium, ⑧ All plates were cultured at 22°C, and single colonies were selected after 2-3 days and stored with numbers. The inhibitory effect on Botrytis cinerea was tested by confronting culture in a plate, and the strain that formed the inhibition z...
Embodiment 2
[0016] Example 2: Determination of the Physiological and Biochemical and 16SrDNA Sequence of Bacillus cereus of the present invention
[0017] Physiological and biochemical test results: zy-1 cultured on NA plate medium at 28°C for 36 hours, the colonies are nearly round, off-white, opaque, large colonies, 3-7mm, irregular edges and diffused, rough particles on the surface like ground glass or Waxy, does not secrete pigment; is a gram-positive bacteria, rod-shaped, 1.0~1.3×3.0~5.0μm, chain, motility, spore oval, terminal or subterminal, spore expansion is not obvious; Cultured by 7% NaCl, contact enzyme positive, glucose fermentation positive, starch hydrolysis positive, nitrate reduction positive, gelatin liquefaction positive, maltose fermentation positive, semi-solid puncture test positive, casein hydrolysis positive, VP test negative, citrate Utilization test is negative, indole test is negative, sucrose fermentation is negative, lactose fermentation is negative, mannitol hyd...
Embodiment 3
[0020] Example 3: Test of Bacillus cereus of the present invention on spore germination and hypha growth of Botrytis cinerea
[0021] 1. Preparation of Bacillus cereus culture filtrate: After strain zy-1 was shaken cultured at 28°C for 3 days, the fermentation broth was centrifuged (12000r / min, 20min), and the supernatant was sterilized with a bacterial filter (0.22μm) to obtain The sterile fermentation broth.
[0022] The spore germination test was carried out by the concave slide method. The Strawberry Botrytis cinerea was attached to a PDA plate and cultured at 23°C for 5 days. After the sporulation, the spores were eluted with sterile water to make a concentration of 10 5 Each / ml spore suspension, suck 50μL of sterile fermentation broth and add it to a 500μL centrifuge tube, and then suck 50μL of the prepared spore suspension to mix the medicine and the spore suspension evenly. Pipette 40 μL of the above mixed solution and drop it on the concave glass slide, then place it in a...
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