Bacillus subtilis 9A and application thereof
A technology of Bacillus subtilis and bacteria agent, which is applied in the field of agricultural microorganisms, can solve safety problems, pollute the environment, pesticide residues and other problems, and achieve the effect of good prospects for the development of biological pesticides
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Embodiment 1
[0029] Example 1 Isolation, screening and identification of Bacillus subtilis 9A
[0030] 1.1 Isolation of bacteria
[0031] Use a sterilized writing brush to swipe on the powdery mildew lesions of melon leaves, and the falling objects fall on the PDA medium plate, and place them in an incubator at 26°C for natural light cultivation. Observe regularly during the cultivation period, purify bacterial colonies with obvious differences in appearance, shape and color, and save them for future use.
[0032] PDA medium formula: potato 200g, glucose 10g, agar 16g, distilled water 1000ml.
[0033] 1.2 Screening and obtaining of antagonistic strain 9A
[0034] Preliminary screening: Antagonistic screening of the bacterial strains isolated and purified in 1.1 was carried out on a PDA plate by using the plate confrontation growth method with a strain of Anthracnose mangoicum as an indicator bacterium, and the strains with antagonistic effects were preserved for future use.
[0035] Re-...
Embodiment 2
[0041] The preparation of embodiment 2 Bacillus subtilis 9A fermented liquid
[0042] (1) Strain activation: Inoculate Bacillus subtilis 9A on solid medium and culture at 25°C-28°C for 1 day;
[0043] (2) Fermented seed culture: Inoculate one loop of the above-mentioned activated bacterial lawn into 100ml of seed culture liquid medium, and vibrate at 180r / m for 12 hours at 25°C-28°C to obtain seed liquid;
[0044] (3) Bacterial agent fermentation: Inoculate the above seed liquid into the bacterial agent fermentation culture medium at an inoculum amount of 0.05% (V / V), and culture at 180r / m at 25°C-28°C for 48 hours to obtain subtilis spores Bacillus 9A bacterial agent (the number of viable bacteria is about 3.0×10 9 cfu / ml).
[0045] The formula of solid medium for strain activation culture is: beef extract 5.0g, peptone 10.0g, NaCl 5.0g, distilled water 1000mL, agar powder 15g, pH7.0-7.2.
[0046] The formula of the liquid culture medium for strain fermented seed culture i...
Embodiment 3
[0048]The preparation of embodiment 3 Bacillus subtilis 9A fermented liquid
[0049] (1) Strain activation: Inoculate Bacillus subtilis 9A on solid medium and culture at 25°C-28°C for 2 days;
[0050] (2) Fermented seed culture: Inoculate a loop of the lawn of the above-mentioned activated strain into 100ml of seed culture liquid medium, and vibrate at 25°C-28°C at 180r / m for 24 hours to obtain a seed liquid;
[0051] (3) Bacterial agent fermentation: Inoculate the above seed liquid into the bacterial agent fermentation culture medium at an inoculum amount of 0.1% (V / V), and vibrate at 180r / m at 25°C-28°C for 72 hours to obtain Bacillus subtilis Bacillus 9A bacterial agent (the number of viable bacteria is about 8.0×10 9 cfu / ml).
[0052] The formula of solid medium for strain activation culture is: beef extract 5.0g, peptone 10.0g, NaCl 5.0g, distilled water 1000mL, agar powder 15g, pH7.0-7.2.
[0053] The formula of the liquid culture medium for strain fermented seed cult...
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