Broad-spectrum antibacterial bacillus amyloliquefaciens strain and application thereof
A technology of Bacillus amyloliquefaciens and Bacillus amyloliquefaciens, which is applied to broad-spectrum antibacterial Bacillus amyloliquefaciens strains and its application fields, can solve the problems of the influence of chemical bactericides and preservatives, and achieve strong broad-spectrum antibacterial activity, long-lasting antibacterial effect, Excellent thermal stability
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Embodiment 1
[0029] Isolation and identification of Bacillus amyloliquefaciens NCPSJ7
[0030] Laiwu City, Shandong Province, China Ginger planting plot soil samples, culture and identification in the laboratory according to the following conditions, can obtain the broad-spectrum antibacterial Bacillus amyloliquefaciens NCPSJ7 of the present invention:
[0031] Isolation and purification of strain NCPSJ7: The soil samples were cultured on beef extract peptone medium by gradient dilution method. Pick a single colony, inoculate it on the beef extract peptone medium by the method of streaking, culture at a constant temperature of 37°C for 1-2 days, pick a single colony and save it.
[0032]The Bacillus amyloliquefaciens NCPSJ7 described in the present invention is identified based on the phylogenetic analysis of its colony morphology and individual morphological characteristics, physiological and biochemical characteristics and 16S rDNA gene sequence. The length of the 16S rDNA gene sequence...
Embodiment 2
[0038] Cultivation and fermentation conditions of the present invention
[0039] (1) Incline culture conditions: Beef extract peptone medium: In aqueous solution, add 0.3% beef extract, 1% peptone, 0.5% NaCl, 2% agar, pH 7.0-7.2, steam at 121°C, 0.1MPa Bacteria for 20 minutes; culture temperature 37 ℃, culture time 2-4 days.
[0040] (2) Seed culture conditions: Seed culture medium: In aqueous solution, add 0.5% glucose, 0.75% yeast extract, 0.75% peptone, (NH 4 ) 2 SO 4 0.5%, NaCl 0.5%, pH 7.0. 121°C, 0.1MPa steam sterilization for 20 minutes; take a piece of fresh strain slant, take a loop of the strain from the slant with an inoculation loop, and transfer it to a container filled with seed medium Incubate in shake flasks at 37°C for 24 hours.
[0041] (3) Fermentation culture conditions: Fermentation medium: In aqueous solution, add 0.5% glucose, 0.75% yeast extract, 0.75% peptone, (NH 4 ) 2 SO 4 0.5%, NaCl 0.5%, pH 7.0. 121℃, 0.1MPa steam sterilization for 20 minu...
Embodiment 3
[0044] Analysis of the antibacterial activity of the broad-spectrum antibacterial thallus and fermentation broth produced by Bacillus amyloliquefaciens NCPSJ7 of the present invention
[0045] Spread the bacterial suspension of indicator bacteria on a suitable medium, among which peach root rot, cucumber wilt, tomato gray mold, jujube anthracnose, pear black spot, pear blue mold, apple brown rot , apple spot defoliation, watermelon wilt pathogenic fungus and yeast spread on potato medium, Staphylococcus aureus and Salmonella paratyphi A spread on beef extract peptone medium, Vibrio parahaemolyticus spread on improved On the beef extract peptone medium (sodium chloride content 3%), place the Oxford cup.
[0046] The fermented liquid that Bacillus amyloliquefaciens NCPSJ7 of the present invention produces is through 4500 revs centrifugation 10min, gets thalline suspension or fermentation supernatant 200uL drop-wise in Oxford cup, and suitable temperature is cultivated (bacterium...
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