Brevundimonas aurantiaca and application thereof to Baitiao codonopsis pilosula
A Brevimonas, yellow technology, applied in the field of microorganisms, can solve the problems of discounted effect, damage, chromosomal aberration of liver disease, etc.
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Embodiment 1
[0034] Example 1 Isolation of Brevundimonas xanthomonas D3 CGMCC No.17101
[0035] Take 5g of soil sample (collected from Zhang County, Gansu Province) into a triangular flask, add 50ml of sterile water, seal it and shake it on a shaker for 20 minutes to make a suspension, absorb 5ml and add it to 100ml of nitrogen-fixing bacteria enrichment medium, 28 Cultivate at ℃, and after 72 hours, absorb 5ml of the bacterial solution and add 100ml of nitrogen-fixing bacteria enrichment medium to continue the cultivation, repeat the cultivation three times, and then isolate a single colony. Separation adopts the plate streaking method and gradient dilution method: the plate streaking method is to use the inoculation loop to dip the bacterial liquid on the solid medium of Ashubei for streaking and separation; the gradient dilution method is to divide the cultured bacterial liquid at 10 -2 、10 -4 、10 -6 、10 -8 Dilution After making a bacterial suspension with sterile water, draw 0.1ml a...
Embodiment 2
[0036] Example 2 Identification of Brevundimonas xanthomonas D3 CGMCC No.17101.
[0037] The isolated and purified D3 strain was identified from the following three aspects.
[0038] 1. Morphological identification
[0039]The D3 isolated in Example 1 was cultured as a single colony and the colonies were observed, mainly including the size, color, transparency, humidity, surface state of the colony (whether it is flat, raised, wrinkled, depressed, etc.), the state of the edge of the colony (whether neat, irregular, radial, etc.). Microscopic examination was performed after staining with biological stains.
[0040] The results showed that the colonies of strain D3 obtained after separation and purification: colony diameter 2-3 mm, colony color yellow, colony opaque, colony surface moist, slightly raised, colony edge neat. Microscopic examination was Gram-negative, with flagella and no spores.
[0041] 2. Separation of 16s rDNA sequence homology
[0042] Cultivate the bacte...
Embodiment 3
[0057] Example 3 Nitrogenase Activity Test of Brevundimonas xanthus D3 CGMCC No.17101
[0058] The Brevundimonas xanthomonas ( Brevundimonas aurantiaca ) D3 CGMCCNo.17101 was used to test the nitrogenase activity, and the acetylene reduction method was used. The specific method is as follows: prepare the suspension of the D3 strain (cells, 1×10 8 ml -1 ), take 1ml and inoculate it in a serum bottle filled with 5ml of semi-solid Ashubei culture medium, and incubate at 28°C for 48h. Replace the cap of the serum bottle with a rubber stopper, draw out 10% of the gas with a sterile syringe, then inject 10% acetylene, and incubate at 28°C for 36h. Use a sterile syringe to extract 0.2ml of the mixed gas from the serum bottle and inject it into the gas chromatograph, measure the amount of ethylene produced, and calculate the nitrogenase activity according to the following formula. N=hxCV / (24.9hst). where N is the nitrogenase activity, that is, the concentration of acetylene produc...
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