A strain of Bacillus subtilis, bacterial agent, bacterial liquid extract and preparation method and application
A technology of Bacillus subtilis and extract, applied in the field of inoculum, bacterial liquid extract and preparation, and Bacillus subtilis field, can solve the problems of enhanced drug resistance of pathogenic bacteria, pollution, ecological environment of pesticide residues, etc.
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[0038] The invention provides the preparation method of the bacterial liquid lipopeptide crude extract described in the above scheme, comprising the following steps:
[0039] 1) Centrifuge the bacterial liquid of Bacillus subtilis ZD01, collect the supernatant, adjust the pH of the supernatant to 2.0, stand at 4°C for 9-16 hours, centrifuge, and collect the precipitate;
[0040] 2) Dissolving the precipitate with methanol, filtering with a 0.22 μm filter, collecting the filtrate to obtain a crude lipopeptide extract.
[0041] In the present invention, the bacterial liquid of Bacillus subtilis ZD01 is firstly centrifuged, the supernatant liquid is collected, the pH value of the supernatant liquid is adjusted to 2.0, and the bacteria liquid is allowed to stand at 4°C for 9 to 16 hours, centrifuged, and the precipitate is collected; the bacterial liquid is preferably It is obtained by the following preparation method: a single colony of Bacillus subtilis ZD01 is inoculated into 2...
Embodiment 1
[0047] Embodiment 1 The effect of different concentrations of Bacillus subtilis ZD01 fermentation broth on potato early blight
[0048] 1. Materials
[0049] 1.1 Test strains
[0050] Bacillus subtilis ZD01 and Alternaria solani were all preserved and provided by the Potato Disease Research Laboratory of Hebei Agricultural University.
[0051] 1.2 Test medium
[0052] LB medium: tryptone 10g·L -1 , Yeast powder 5g L -1 , Sodium chloride 10g L -1 pH 7.0-7.2.
[0053] LBA medium: tryptone 10g·L -1 , Yeast powder 5g L -1 , Sodium chloride 10g L -1 , agar 20g·L -1 .
[0054] PDA medium: potato 200g·L -1 , Glucose 20g L -1 , agar 20g·L -1 .
[0055] Oat medium: oat 30g·L -1 , agar 15g·L -1 .
[0056] Tomato medium: 125g·L -1 , agar 15g·L -1 , pH 7.0.
[0057] 2. Method
[0058] 2.1 Preparation of fermentation broth
[0059] Take the ZD01 strain stored in the refrigerator at -80 °C, streak it on LBA medium, stand at 37 °C for 24 hours, pick a single colony and in...
Embodiment 2
[0065] Example 2 The effect of crude extracts of Bacillus subtilis ZD01 lipopeptide with different concentrations on potato early blight
[0066] 1. Extraction of lipopeptides
[0067] 1) Preparation of fermented seed liquid: take ZD01 strain stored in -80°C refrigerator, streak it on LBA medium, culture at 37°C for 24h, pick a single colony and inoculate it in a conical flask containing 100mL LB liquid medium at 37°C , 200rmp shaking culture for 72h to obtain fermented seed liquor.
[0068] 2) Extraction of lipopeptide compounds: acid precipitation was adopted for the extraction of crude lipopeptide extracts, and the fermentation broth obtained during the fermentation of ZD01 strain was centrifuged at 4° C. 8000 rmp for 20 min, the supernatant was collected, and slowly added to the supernatant. Adjust pH to 2.0 with 6 mol / L HCl, overnight at 4 °C; then centrifuge at 8000 rmp for 20 min at 4 °C, collect the precipitate, dry the precipitate in a ventilated place, and add 30 ml...
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