Bacillus subtilis microbial agent and preparation method and application thereof
A technology of Bacillus subtilis and bacteria agent, applied in the field of microorganisms, can solve the problems of loss of biodiversity, plant death, increase of water and soil erosion, etc., and achieve the effect of promoting growth and reproduction and good antibacterial ability
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Embodiment 1
[0023] The preparation of embodiment 1 bacterial agent
[0024] (1) Strain activation: inoculate the low-temperature preserved Bacillus subtilis JQ-0101D strain on LB solid medium, and culture at 36°C for 36h; pick a single colony and transfer it to LB slant medium, and culture at 36°C for 20h , wash the bacteria on the surface of the culture medium with sterile water as the inoculum.
[0025] (2) Preparation of first-level seed liquid: insert step into seed culture medium according to the inoculum size of 10% (volume percentage)
[0026] (1) The prepared inoculum was cultured on a shaking table for 20 h at a rotational speed of 220 rpm and a temperature of 36° C., as a primary seed solution.
[0027] 3) Secondary seed liquid preparation: according to the inoculum size of 3% (volume percentage), the step of inserting into the seed medium
[0028] (2) The first-grade seed solution was prepared with an aeration rate of 1.0vvm, a stirring speed of 220rpm, and a culture time of ...
Embodiment 2
[0033] Example 2 Antibacterial Detection
[0034] In a plate with a partition in the middle, pipette 200 μL of the prepared 10 9 CFU / ml of Bacillus subtilis fermentation broth on LB medium, spread evenly with a sterilized coating rod, and wait for its surface to dry; while inoculating three kinds of pathogenic bacteria on PDA medium respectively, place the plate at 25°C under cultivation. Do 3 parallel groups, use LB medium coated with sterile saline as a control, and observe the antibacterial effect after 72 hours.
[0035] The experimental results are shown in Table 1
[0036] Types of pathogenic bacteria
[0037] Inhibitory effect
[0038]
[0039] Note: + the inhibitory effect is weak, the inhibition rate of the colony radius is less than 10%, which affects the growth of fungal mycelia; ++ the antibacterial effect is obvious, the inhibition rate of the colony radius is between 10% and 20%; The rate is more than 20%; the average of three repetitions is taken.
[00...
Embodiment 3
[0041] Example 3 Effect on Acidobacteria (family) (qualitative experiment)
[0042] Taking Citrobacter as the representative of Agrobacterium (family), perform the following two operations respectively, and do 3 groups in parallel:
[0043] 1) Inoculate into MRS basal medium and shake directly, after 8 hours, detect OD600=0.6;
[0044] 2) Take the JQ-0101D fermentation broth, filter out the hyphae, and replace the water in the MRS basal medium. Other components remain unchanged. The improved medium is inoculated with Citrobacter at a 5% inoculation amount, and then the shaker 8h, detect OD600=0.8.
[0045] It can be seen that the fermentation product of Bacillus subtilis JQ-0101D can promote the reproduction of acid bacteria.
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