Microbial inoculant for improving disease resistance and promoting growth of plants
A technology of microbial agent and microbial strain, applied in the direction of plant growth regulators, plant growth regulators, microorganisms, etc., can solve the problems of soil microbial zone interference, enhanced drug resistance of pathogenic bacteria, difficulty in functioning normally, etc., to promote growth , growth enhancement, good market prospects
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Embodiment 1
[0032] Preparation of bacteria (Bacillus subtilis)
[0033] (1) Bacillus subtilis Isolation of EBC7: Collect healthy “Jinyou 10” cucumber roots from the greenhouse in Xiqing District, Tianjin; rinse with running water for 1 hour, soak in 70% ethanol for 1 minute, and disinfect in 2.5% NaClO for 6 minutes , rinse with sterile water and set aside;
[0034](2) Transfer the sterilized tissue obtained in step (1) to a sterile mortar and add PBS buffer solution for grinding, take the grinding juice and serially dilute, apply 100 μl each on the NA plate, and place it at 28°C Dark culture, each treatment repeated 3 times;
[0035] (3) Pick a single colony on the NA plate, streak and purify it, and obtain the Bacillus subtilis strain; then inoculate the Bacillus subtilis in the NB culture medium, shake it at 150 rpm / min for 24 hours, and then inoculate the obtained Bacillus subtilis Store the bacillus solution in 15% glycerol at -80°C for later use; NA medium: 10g peptone, 5g NaCl,...
Embodiment 2
[0037] Bacteria (Bacillus subtilis) are stored in 15% glycerol at -80°C, and inoculated on NA solid medium for activation and culture when in use; after activation, take a ring of bacterial lawn and inoculate it in a triangular flask containing NB medium for seed culture. After shaking at 150 rpm / min for 24 h, inoculate 3% of the inoculum in a fermentation bottle and ferment for 48 h to obtain a content of 1.5×10 10 The fermentation broth contains vegetative bodies and / or spores of Bacillus and antibacterial metabolites.
[0038] NA medium: peptone 10g, NaCl 5g, beef extract 3g, agar powder 12g, water up to 1000ml, pH7.0;
[0039] NB medium (seed culture medium): peptone 10g, NaCl 5g, beef extract 3g, water up to 1000ml, pH7.0;
[0040] Fermentation medium: soybean powder 10.00g, corn starch 5.00g, yeast powder 0.50g, KH 2 PO 4 0.35g, CaCl 2 0.20g, add water to 1000ml, pH7.0.
Embodiment 3
[0042] Bacteria (Bacillus subtilis) are stored in 15% glycerol at -80°C, and inoculated on NA solid medium for activation and culture when in use; after activation, take a ring of bacterial lawn and inoculate it in a triangular flask containing NB medium for seed culture. After shaking at 150 rpm / min for 24 h, inoculate 3% of the inoculum in the fermentation bottle and ferment for 48 h to obtain a content of 4.5×10 10 Fermentation broth (containing vegetative and / or spores of Bacillus and antibacterial metabolites).
[0043] NA medium: peptone 10g, NaCl 5g, beef extract 3g, agar powder 12g, water up to 1000ml, pH7.0;
[0044] NB medium: peptone 10g, NaCl 5g, beef extract 3g, water up to 1000ml, 7.2;
[0045] Fermentation medium: soybean powder 10.00g, corn starch 5.00g, yeast powder 0.50g, KH 2 PO 4 0.35g, CaCl 2 0.20g, add water to 1000ml, pH7.2.
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