Profitable soil microorganism mixed fermentation method
A mixed fermentation and microbial technology, which is applied in the field of mixed fermentation of Issaccharomyces orientalis, Bacillus licheniformis, and soil beneficial strain Paecilomyces lilacinus, can solve the problems that the fermentation process has no special instructions, and achieve simple and efficient fermentation methods and formulas Simple, reduce the effect of chemical fertilizer and pesticide application
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Embodiment 1
[0028] Example 1 Mixed medium formulation optimization
[0029]With beef extract 3g / L, peptone 12g / L, glucose 20g / L, potassium dihydrogen phosphate 2g / L, and zinc sulfate 0.1g / L as the basal medium, the culture conditions were initial pH 6.5, 28°C, 180rpm shaking 3d. Three strains of bacteria could be observed through microscopic examination and medium plate coating, figure 1 It is the colony morphology of the three strains of bacteria on the basal medium, indicating that the basal medium is suitable for the cultivation of the three strains of bacteria.
[0030] Substituting sucrose, maltose, and soluble starch for glucose in the basal medium with the addition of 20g / L respectively, investigated their influence on the effective number of viable bacteria of the three strains of mixed culture, as figure 2 As shown, when soluble starch was used as carbon source, the total effective number of viable bacteria of the three strains was the highest, followed by sucrose, maltose and...
Embodiment 2
[0041] The culture effect comparison of embodiment 2 bacterial strain specific medium and mixed fermentation medium
[0042] After the activation of Issaccharomyces orientalis, scrape a ring of bacteria from the slope and put it in a triangular flask filled with normal saline, shake it well and dilute to OD 600 0.2, with 10% of the inoculum amount, inoculated respectively in the following three liquid culture mediums known to cultivate Issahia orientalis and the mixed fermentation liquid medium of the present invention, cultivated at 28°C and 180rpm for 48h, and took samples to detect effective viable bacteria number.
[0043] The substratum of three kinds of known oriental Issa yeasts of table 3 and the formula of mixed fermentation medium of the present invention
[0044]
[0045] After activation of Bacillus licheniformis, scrape a ring of bacteria from the inclined plane and put it in a triangular flask filled with normal saline, shake well and dilute to OD 600 0.2, i...
Embodiment 3
[0052] The quality control of embodiment 3 mixed fermentation inoculum size
[0053] In order to control the effective number of viable bacteria in the initial inoculation of various bacteria in the mixed fermentation system, so that the initial effective number of viable bacteria of various bacteria is within the known controllable range, the effective number of viable bacteria in the seed solution and the OD 600 The relationship between the values, the number of effective viable bacteria and OD of the three bacterial seed solutions were made 600 Standard curve between values.
[0054] After inoculation of Paecilomyces lilacinus, culture 4-6 days to collect spores, wash the spores with normal saline to prepare spore suspension, adjust the spore suspension to OD with sterile water 600 was 0.2, inoculated the spore suspension into the liquid medium with 10% inoculum, cultivated it at 28°C and 180rpm for 2 days, and prepared Paecilomyces lilacinus seed liquid, which was counted...
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