Bacillus subtilis, microbial agent and application of bacillus subtilis and microbial agent to fermentation compost maturity
A technology of Bacillus subtilis and microbial inoculum, which is applied in the field of agricultural biology, can solve the problem that the efficiency of fermentation and decomposing needs to be improved, and achieve the effects of improving the efficiency of fermentation and decomposing and improving the efficiency.
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Embodiment 1
[0047] This example is used to illustrate the cultivation of Bacillus subtilis and the preparation of microbial inoculum of the present invention.
[0048] The slant bacterial classification of Bacillus subtilis (Bacillus subtilis) with preservation number is CCTCCNO: M2013496 on LB plate (containing the peptone of 10g / L, the yeast extract of 5g / L, the NaCl of 5g / L and the agarose of 3g / L, pH value 7.4, the same below), and cultured at 37°C for 15 hours, picked a single colony from the plate, and inoculated it in 25mL LB liquid medium (containing 10g / L peptone, 5g / L yeast extract and 5g / L of NaCl, pH value 7.4, hereinafter the same), cultivated on a shaker at 250r / min at 37°C for 15 hours to obtain first-class seeds, and added 20mL of first-class seeds to 500mL of LB liquid medium, Cultivate on a shaker at 250r / min at 37°C for 15 hours to obtain secondary seeds, add 2L of secondary seeds to 50L of LB liquid medium, stir at 250r / min at 37°C and 10m 3 / h ventilation for 15 hour...
Embodiment 2
[0082] This example is used to illustrate the cultivation of Bacillus subtilis and Bacillus licheniformis of the present invention and the preparation of microbial inoculum.
[0083] The slant bacterial classification of Bacillus subtilis (Bacillus subtilis) with preservation number is CCTCCNO: M2013496 on LB plate (containing the peptone of 10g / L, the yeast extract of 5g / L, the NaCl of 5g / L and the agarose of 3g / L, pH value 7.4, the same below), and cultured at 37°C for 15 hours, picked a single colony from the plate, and inoculated it in 25mL LB liquid medium (containing 10g / L peptone, 5g / L yeast extract and 5g / L of NaCl, pH value 7.4, hereinafter the same), cultivated on a shaker at 250r / min at 37°C for 15 hours to obtain first-class seeds, and added 20mL of first-class seeds to 500mL of LB liquid medium, Cultivate on a shaker at 250 r / min at 37° C. for 15 hours to obtain secondary seeds of Bacillus subtilis.
[0084] The slant bacterial classification of Bacillus lichenif...
Embodiment 3
[0088] The microbial inoculum was prepared according to the method of Example 2, except that 1.2 L of secondary seeds of Bacillus subtilis and 1.2 L of secondary seeds of Bacillus licheniformis were mixed, and then tertiary expansion culture was carried out.
[0089] In this embodiment, the CFU ratio of the Bacillus subtilis to the Bacillus licheniformis in the culture medium was determined to be 1:0.9 by using the nucleic acid quantification method based on 16SrDNA.
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Abstract
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