Bacillus amyloliquefaciens WS-8 and application thereof
A technology for dissolving starch spores and bacilli is applied in the field of agricultural microorganisms and biology to achieve the effects of improving tomato quality, having broad market application prospects and reducing pesticide residues
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[0017] Example 1: Isolation and identification of Bacillus amyloliquefaciens WS-8
[0018] Infected and healthy vegetable leaves were collected from tomato and cucumber planting sheds in Dahe Town, Luquan, Shijiazhuang, and cultivated in our laboratory according to the following conditions to obtain the broad-spectrum antibacterial Bacillus amyloliquefaciens WS-8 of the present invention:
[0019] Isolation and purification of strain WS-8: The healthy samples and diseased leaves after removal of rot and mold were diluted by gradient method, and 10 grams of leaves were put into phosphate buffer solution, shaken at room temperature at 200 r / min for 30 minutes, and ultrasonically treated for 5 minutes. The bacterial suspension was coated on a plate, and a single colony was picked for purification by streaking.
[0020] Screening of antagonistic strains WS-8: The purified single colony was used for primary screening and re-screening of resistant strains with tomato leaf mold and...
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[0027] Example 2: Determination of antibacterial spectrum of Bacillus amyloliquefaciens WS-8
[0028] The strains of the present invention are airborne fungal diseases: pear ring rot, pear black spot, cucumber gray mold, tomato leaf mold, tomato gray mold, tomato early blight, tomato soot; soil-borne fungal diseases : blight, fusarium wilt, verticillium wilt, vine blight; food-borne bacteria: Staphylococcus aureus, the antibacterial spectrum of various pathogenic strains was measured, and the results are shown in Table 2.
[0029] Table 2 Bacteriostatic spectrum of Bacillus amyloliquefaciens subsp. WS-8
[0030]
[0031] Note: The strength of antagonism is indicated by "+''. "+": weak, the width of the inhibition zone is less than 1.5cm; "++'': medium strength, the width of the inhibition zone is between 1.5cm and 3.0cm;" +++": very strong, the width of the antibacterial zone is greater than 3.0cm.
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[0032] Example 3: Culture and fermentation conditions of Bacillus amyloliquefaciens WS-8
[0033] Bacterial strains with strong antibacterial effect were purified, and beef extract peptone medium was used, which was stored on a solid slant; a ring was picked from the slant culture and inoculated into the beef extract peptone liquid medium for overnight culture at 37°C as a bacterial seed. Bottle ready to use.
[0034] Fermentation culture conditions: Fermentation medium: starch 0.7%, bean cake flour 1.6%, sucrose 0.2%, yeast extract 0.08%, peptone 0.05%, ammonium sulfate 0.15%, potassium dihydrogen phosphate 0.05%, magnesium sulfate 0.03%, manganese sulfate 0.01% %, sodium chloride 0.02%, calcium carbonate 0.1%, pH 7.3-7.5, the above are all mass percentages. Sterilize at 121°C for 20 minutes, and after cooling, add the seed liquid to the fermentation broth with an inoculum amount of 1.5-2%, 600mL / 3L conical flask, rotate at 180 rpm, temperature 30°C, and cultivate for 48 h...
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