Microbial inoculant and culture medium, and preparation methods thereof
A microbial agent and substrate technology, applied in the field of cultivation, can solve the problems of lack of comprehensive utilization means of agricultural waste and lack of new cultivation substrate raw materials, etc., and achieve the effects of increasing fruit and vegetable yield, loose texture, and promoting production and development
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[0023] Wherein, the preparation method of the microbial bacterial agent may include: respectively inoculating Bacillus subtilis, Pseudomonas stutzeri ACCC01975 and Micrococcus ACCC19875 with the preservation numbers of CGMCC No. 12318 in the medium for culturing. Preferably, the viable counts of Bacillus subtilis, Pseudomonas stutzeri ACCC01975 and Micrococcus ACCC19875 with the preservation numbers CGMCC No. 7 -10 11 cfu, more preferably 10 8 -10 9 cfu. During the cultivation process, the concentration of live bacteria can be obtained by conventional methods, such as hemocytometer or OD value observation.
[0024] The cultured bacterial solution can be directly used as a microbial bacterial agent. Preferably, the bacterial solution is further processed into a microbial bacterial agent in a dosage form that is more convenient for storage through steps including aseptic filtration and freeze-drying. Wherein, the culture conditions of the strains are not particularly limited...
Embodiment 1
[0040] This example is used to illustrate the preparation of the microbial inoculum of the present invention.
[0041] Inoculate Bacillus subtilis with preservation number CGMCC No.12318 into LB medium (containing 10g / L of tryptone, 5g / L of yeast extract and 10g / L of NaCl), at 30°C and 200 rpm Incubate with shaking. Gained bacterium liquid is the microbial inoculum V1 of the present embodiment, and in every gram of this microbial inoculum, the number of live bacteria is 10 9 cfu.
Embodiment 2
[0043] This example is used to illustrate the preparation of the microbial inoculum of the present invention.
[0044] (1) The preservation number is CGMCC No.12318 Bacillus subtilis, the product number is ACCC01975 Pseudomonas stutzeri and the product number is ACCC19875 Micrococcus respectively inoculated into beef extract peptone medium (containing 20g / L of peptone , 5g / L beef extract and 5g / L NaCl), cultured at 37°C and 180 rpm shaking, samples were taken during the culture and observed by direct microscope counting until the preservation number was CGMCC No .12318 of Bacillus subtilis, Pseudomonas stutzeri ACCC01975 and Micrococcus ACCC19875 respectively reached 10 8 cfu / gram.
[0045] (2) Mix the Bacillus subtilis obtained in (1), Pseudomonas stutzeri ACCC01975 and Micrococcus ACCC19875 in proportion, so that in the obtained microbial agent V2, the preservation number is the Bacillus subtilis of CGMCC No.12318 , Pseudomonas stutzeri ACCC01975 and Micrococcus ACCC19875 ...
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