Bacillus subtilis and application thereof
A technology of Bacillus subtilis and microbial strains, applied in the field of microorganisms, can solve problems such as undeveloped and impact on rice quality, achieve strong antibacterial activity, and promote the effect of increasing crop production
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Embodiment 1
[0022] Embodiment 1: the fermentation method of bacillus subtilis T500
[0023] The fermentation of Bacillus subtilis T500 comprises the steps:
[0024] 1) Strain activation: transfer Bacillus subtilis T500 to YPGA slant medium, and culture at 28°C for 48 hours;
[0025] 2) Seed culture: transfer the activated strain in step 1) into YPG culture solution, culture at 120 rpm, 28° C. for 48 hours.
[0026] 3), fermentation culture: in step 2), the content of bacteria in the seed tank reaches 10 10 When cfu / ml, put the fermentation hydraulic pressure in the seed tank into the fermenter for fermentation, the fermentation temperature is 28-32°C; the rotation speed: 80-150 rpm; Detect every 2 hours, observe the bacteria content and measure the pH of the fermentation broth. After 90% of the bacteria turned into spores, the detected bacteria content reached 2×10 10 cfu / ml out of the tank; after out of the tank, add benzoic acid preservative to the fermentation broth according to 2%...
Embodiment 2
[0030] Embodiment 2: Bacteriostasis of T500 on plant diseases
[0031] Determination of antibacterial activity against rice sheath blight: connect 9mm rice sheath blight bacteria cake to the center of the PDA medium plate, place 6 mm sterile filter paper sheets at three points, and absorb 10 μL of Bacillus subtilis fermentation broth on the filter paper cultured in an incubator at 28°C. The treatment without Bacillus subtilis was set as the control, and each treatment was repeated 3 times. When the plate was covered with pathogenic bacteria in the blank control, the diameter of the inhibition zone was measured and the inhibition rate was calculated. The method for measuring the antibacterial activity of Fusarium graminearum, Aspergillus flavus and Aspergillus niger is the same as that of Aspergillus oryzae. Determination of the antibacterial activity of rice smut fungus: Inoculate the rice smut fungus into PDA liquid medium, culture it on a shaker at 140 r / min, 28 ℃ for 5-7 d...
Embodiment 3
[0035] Embodiment 3, the compatibility of bacillus subtilis and tricyclazole
[0036] Under sterile conditions, Bacillus subtilis T500 was inserted into the NB culture medium and cultured at 28°C for 48 h on a shaker at 140 r / min. Weigh an appropriate amount of 20% tricyclazole, and prepare to contain tricyclazole concentrations of 100 μg / mL, 500 μg / mL, 1000 μg / mL, 2000 μg / mL, 3000 μg / mL, 4000 μg / mL, 5000 μg / mL potato plate and potato plate without tricyclazole, put filter paper at fixed points, and the filter paper contains 10 μL of cultured T500 fermentation broth, and use the filter paper in the potato plate without tricyclazole as a control , Observe and record the colony growth after culturing at 28°C for 2 days.
[0037] The results are shown in Table 3, indicating that the growth of Bacillus subtilis T500 was not affected on the medium with a tricyclazole concentration of 4000 μg / mL. When the medium contained 5000 μg / mL of tricyclazole, the colonies were smaller and t...
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