Bacillus subtilis formulation for preventing and controlling soil-borne disease and method of preparing the same
A technology for Bacillus subtilis and soil-borne diseases, applied in the fields of botanical equipment and methods, fungicides, biocides, etc. Ecological environment, broad antibacterial spectrum, no environmental pollution effect
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Embodiment 1
[0024] The fermentation steps of Bacillus subtilis B579 are as follows:
[0025] Connect the Bacillus subtilis strain B579 (Bacillus subtilis) to the glass plate A containing the first medium, and culture it at 37°C for 12 hours to obtain the strain B579;
[0026] The activated strain B579 was inoculated into a 250 mL Erlenmeyer flask containing 30 mL of the second medium, and fermented for 12 hours to obtain a seed solution. The culture conditions are better when the inoculation amount is 1-10%, the inoculation time is 4-16 hours, and the rotational speed is 100-200r / min. In particular, the inoculation amount is 1%, the inoculation time is 12 hours, and the rotational speed is 180r / min.
[0027] Transfer the seed solution at 3% into a 7L fermenter equipped with the third culture medium, the temperature is 37°C, the inoculum size, the fermentation time is 22-26 hours, and the rotation speed is 100-200r / min.
[0028] Wherein, the first medium formula is: potato 200g / L, agar 2...
Embodiment 2
[0043] The preparation steps of Bacillus subtilis B579 bacterial agent (hereinafter referred to as biocontrol bacteria B579):
[0044] 1) Strain activation: Activate the Bacillus subtilis strain B579 stored at -6°C on a PDA plate medium (30°C), pick a single colony and multiply it on a PDA slant medium for use;
[0045] 2) Preparation of seed liquid: make LB liquid medium according to the conventional method, fill 30mL of LB liquid medium in a 250mL Erlenmeyer flask, and sterilize with high-pressure damp heat. After the temperature drops to room temperature, insert each bottle into step 1) A ring of the above-mentioned activated B579 strain was cultured on a shaking table with a rotation speed of 180rpm, and cultured at 37°C for 12h, and the obtained can be used as a seed solution;
[0046] 3) Preparation of B579 fermentation medium: In a 7L fermenter, add 30g of yeast extract, 55g of soluble starch, 30g of glucose, 70g of soybean cake powder, 45g of beef, and 63g of corn flou...
Embodiment 3
[0052] Application of biocontrol bacteria B579 inoculum in the control of plant soil-borne diseases.
[0053] The bacillus subtilis B579 liquid preparation (content: viable count is 5.0 * 10 8 ~8.0×10 10 cfu / ml), some experiments were done in Tianjin Plant Protection.
[0054] ①Effects of soaking seeds of Bacillus subtilis B579 for different times on cucumber germination.
[0055] Soak the cucumber seeds with 10mL of the fermentation broth of bacteria B579, then place the seeds in sterilized plates covered with double-layer filter paper, inject 5ml of sterile water into the plates every day to moisturize and accelerate germination (25°C), and investigate the germination rate of cucumbers for 36 hours and buds grow.
[0056] ② Control experiment of biocontrol bacteria B579 on cucumber seedling blight (pot experiment)
[0057] The test was carried out under greenhouse conditions (25°C), and the nutrient pots were filled with sterilized soil for use. Three treatments were se...
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