A kind of bacillus licheniformis and microbial agent and their application in the prevention and treatment of plant cropping disease
A technology of bacillus licheniformis and microbial agents, applied in the field of agricultural biology, to achieve the effect of improving the level of prevention and control
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Embodiment 1
[0042] This example is used to illustrate the cultivation of Bacillus licheniformis of the present invention and the preparation of microbial inoculum.
[0043] The slant strain of Bacillus licheniformis (Bacillus licheniformis) with the preservation number of CCTCC NO: M2013495 was placed on an LB plate (containing 10 g / L of peptone, 5 g / L of yeast extract, 5 g / L of NaCl and 3 g / L of agar sugar, 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 of LB liquid medium (containing 10g / L peptone, 5g / L Yeast extract and 5g / L NaCl, pH value 7.4, hereinafter the same), cultivated on a shaker at 250r / min at 37°C for 15 hours to obtain primary seeds, and added 20mL of primary seeds to 500mL of LB liquid medium In 250r / min shaker at 37°C for 15 hours to obtain secondary seeds, 2L of secondary seeds were added to 50L of LB liquid medium, stirred at 250r / min at 37°C and 10m 3 / h ventilation for 15 hours to ob...
Embodiment 2
[0087] This example is used to illustrate the cultivation of Bacillus licheniformis and Bacillus subtilis of the present invention and the preparation of microbial inoculum.
[0088] The slant strain of Bacillus licheniformis (Bacillus licheniformis) with the preservation number of CCTCC NO: M2013495 was placed on an LB plate (containing 10 g / L of peptone, 5 g / L of yeast extract, 5 g / L of NaCl and 3 g / L of agar sugar, 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 of LB liquid medium (containing 10g / L peptone, 5g / L Yeast extract and 5g / L NaCl, pH value 7.4, hereinafter the same), cultivated on a shaker at 250r / min at 37°C for 15 hours to obtain primary seeds, and added 20mL of primary seeds to 500mL of LB liquid medium , cultivated on a shaker at 250 r / min at 37°C for 15 hours to obtain secondary seeds of Bacillus licheniformis.
[0089] The slant strain of Bacillus subtilis (Bacillus subtilis...
Embodiment 3
[0093] The microbial agent was prepared according to the method of Example 2, except that 1.2 L of secondary seeds of Bacillus licheniformis and 1.2 L of secondary seeds of Bacillus subtilis were mixed, and then tertiary expansion cultivation was carried out.
[0094] In this embodiment, the CFU ratio of the Bacillus licheniformis and the Bacillus subtilis in the culture medium measured by the nucleic acid quantification method based on 16S rDNA is 1:0.9.
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