A strain of Bacillus subtilis bj-1 for biocontrol and its application
A technology of Bacillus subtilis and fungi, which is applied in the field of agricultural microorganisms to achieve the effects of strong stability, fast reproduction and storage resistance
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Embodiment 1
[0026] Example 1: Isolation, screening and fermentation of Bacillus subtilis BJ-1
[0027] 1. Isolation, identification and determination of antibacterial spectrum
[0028] The microorganism of the present invention is a Bacillus subtilis with strong antagonistic activity isolated from a naturally polluted tomato and oat plate in the Hubei Provincial Key Laboratory of Crop Disease Detection and Safety Control, Huazhong Agricultural University, Wuhan City, Hubei Province. Separation and purification refer to the plant disease research method (Fang Zhongda, 1998). The method is to pick a Bacillus subtilis colony from a naturally polluted plate, dilute it with 5ml sterile water, dip a small amount of diluent with an inoculation needle and draw it on a PDA plate. Line, select the desired characteristic single colony, after 3 times of the same streak culture, finally get the pure Bacillus subtilis strain. The applicant named it BJ-1, and finally identified the biocontrol bacterial...
Embodiment 2
[0039] Embodiment 2: Bacillus subtilis BJ-1 fermentation filtrate is to the inhibitory effect and teratogenic effect of blast fungus mycelia growth
[0040] 1. Inhibitory effect of Bacillus subtilis BJ-1 fermentation filtrate on the growth of blast fungus mycelia
[0041] After adding the Bacillus subtilis fermentation broth filtrate to PDB and mixing it into different concentrations (v / v, 0, 0.5%, 1%, 2.5%, 5%, 10%, 20%), the rice blast with a diameter of 5mm was beaten Mycelia blocks were inoculated in 75mL of PDB culture medium containing different concentrations of fermentation broth filtrates, treated without adding fermentation filtrates as a control, placed at 28°C and 180rpm for shaking culture for 7 days, and then used pre-dried and weighed qualitative filter paper (diameter 10cm) filter the mycelium, and place it in a 60°C drying oven to dry to constant weight, and weigh the dry weight of the mycelium with an electronic balance with an accuracy of one ten-thousandth ...
Embodiment 3
[0049] Example 3: Inhibition of Bacillus subtilis BJ-1 fermentation filtrate on blast fungus spore germination and appressorium formation
[0050] The Bacillus subtilis BJ-1 fermentation broth filtrate was diluted with 0.025% Tween to different concentrations (v / v, 1%, 2%, 5%, 10%, 20%, 40%) and then mixed with Tween 20 (final concentration 0.025%) at a concentration of 1×10 6 The equal volumes of the blast fungus spore liquid / ml are mixed, so that the concentration of the fermented filtrate contained in the spore liquid is respectively (v / v, 0.5%, 1%, 2.5%, 5%, 10%, 20%). Plague has a spore concentration of 5×10 5 spores / ml. The above-mentioned blast fungus spore liquid containing different concentrations of Bacillus subtilis fermentation broth filtrate was inoculated onto hydrophobic slides respectively, 4 drops per slide, 10 μl per drop. After inoculation, place them under constant temperature and dark conditions at 28°C for 18 hours of moisturizing culture, and then dir...
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