Bacillus methylotrophicus UTM401 and applications thereof
A technology of bacillus and methyl nutrition, applied in the application, bacteria, fungicides and other directions, to achieve the effect of low production cost, excellent performance and inhibition of plant pathogenic bacteria
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Embodiment 1
[0032] Example 1 Isolation and Identification of Methylotrophic Bacillus UTM401
[0033] Weigh 10g of sludge decomposed compost sample (collected from the compost fermentation workshop of Dadi Lvyuan Environmental Protection Technology (Beijing) Co., Ltd.) and place it in a 100ml sterile water 250ml triangular flask with 10 small glass beads, and shake it at a constant temperature at 40°C Shake on the bed for 1 hour, then let it stand for 30 minutes, in the ultra-clean bench, absorb 1ml of the supernatant, and transfer it to 100ml of sterilized acclimation medium (3g of beef extract, 3g of yeast extract, 8g of peptone, chloride Sodium 3g, soluble starch 10g, 1000ml distilled water), enrichment and acclimatization culture for 3 days on a constant temperature shaker at 40°C with a rotation speed of 160r / min. In the same way, draw 1ml of the enrichment solution again, and subculture for another 3 generations under the same conditions.
[0034] Take 1mL of the acclimatized bacter...
Embodiment 2
[0037] The preparation of embodiment 2 strain UTM401 fermented seed liquid (1)
[0038] Transfer the slant surface of the production strain UTM401 of the present invention stored at 4°C to the NA medium plate, cultivate it at 30°C until it is thick, and then use a sterile glass spatula to transfer all of it to a 5000ml triangle containing 1000ml LB liquid medium In the bottle, culture on a shaker at 37°C with a rotation speed of 160r / min for 2 days. At this time, the OD 600 2.202, stop the cultivation, this is the primary fermentation seed liquid.
[0039] The primary fermentation seed liquid was inoculated into a fermentation seed tank containing LB liquid medium at 5% of the volume ratio of the liquid medium for fermentation. The fermentation conditions are temperature 37°C, intermittent stirring and ventilation, stirring once every 1 hour, stirring for 5 minutes each time, stirring speed 150r / min, ventilation once every half an hour, 5 minutes each time, ventilation rate 1...
Embodiment 3
[0041] The preparation of embodiment 3 strain UTM401 fermented seed liquid (2)
[0042] Transfer the slant surface of the production strain UTM401 of the present invention stored at 4°C to the NA medium plate, cultivate it at 30°C until it is thick, and then use a sterile glass scraper to transfer all of it to a 5000ml triangle containing 2000ml LB liquid medium In the bottle, culture on a shaker at 30°C with a rotation speed of 150r / min for 3 days. At this time, the OD 600 It is 2.084, stop cultivating, this is the primary fermentation seed liquid.
[0043] The primary fermentation seed liquid was inoculated into the fermentation seed tank containing LB liquid medium according to 10% of the volume ratio of the liquid medium to carry out fermentation culture. Fermentation conditions are temperature 30°C, intermittent stirring and ventilation, stirring once every 1 hour, stirring for 10 minutes each time, stirring speed 120r / min, ventilation once every half an hour, 5 minutes ...
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