Sphingobium abikonense HP10 and application thereof
A technology of Sphingobacterium, pathogenic bacteria, applied in the field of microbiology
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Embodiment 1
[0031] Example 1 strain isolation
[0032] Weigh 10g of soil sample (collected from Lanzhou City, Gansu Province) into a triangular flask, add 100mL of sterile water, let stand for 20min, 28°C, speed 200r min -1 , shaken for 30 min to form a suspension. Draw 5mL of the suspension and add it to 100mL Ashby liquid medium, at 28°C, at a speed of 180r min -1 After 72 hours of enrichment culture, the conventional gradient dilution method was used to prepare 10 -4 、10 -5 , 10 -6 、10 -7 After diluting the gradient bacterial suspension, draw 0.1mL of the bacterial suspension and spread it on the Ashby medium plate, repeat each gradient 3 times, place at a constant temperature of 28°C, and incubate for 5d to 7d. Repeat the above separation operation 3 times. After the isolated colonies grow out, continue to purify the strains on the Ashby medium plate by streaking the plate. Obtain a colony diameter of 0.5 ~ 1mm, the color of the colony is milky white, the colony is opaque, the ...
Embodiment 2
[0033] Example 2 strain identification.
[0034] The isolated and purified strains were identified from the following three aspects.
[0035] 1. Morphological identification
[0036] The HP10 isolated in Example 1 was cultured for a single colony and observed for colonies, mainly including the size, color, transparency, surface state (whether flat, raised, wrinkled or sunken, etc.) and edge state (whether neat, irregular or Radial, etc.) and late growth state. Microscopic examination was performed after staining with biological stains.
[0037] The results showed that the colonies of the strain HP10 obtained after separation and purification: the colony diameter was 0.5-1 mm, the color of the colony was milky white, the colony was opaque, the surface was wrinkled, the edge was irregular, and the later stage was brown. Microscopic examination was Gram-negative, short rod-shaped, and non-spore-forming.
[0038] 2.16s rDNA sequence homology separation
[0039] The bacterial ...
Embodiment 3
[0056] Nitrogenase activity test of embodiment 3 Sphingobacterium HP10
[0057] Nitrogenase activity test was carried out on the Sphingobacterium HP10 obtained by the isolation and purification of Example 1, and the acetylene reduction method was adopted. The specific method is as follows:
[0058] A ring of Sphingobacterium HP10 was inoculated into a serum bottle containing 5 mL of semi-solid Ashby medium, and cultured at 28°C for 48 hours. Replace the cap of the serum bottle with a rubber stopper in the aseptic table, draw out 1mL of gas with a sterile syringe, then inject 1mL of acetylene, seal it, 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.
[0059] N=hxCV / (24.9hst). Wherein N is the concentration nmol (C 2 h 4 )·h -1 ·mL -1That is nitrogenase activ...
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Abstract
Description
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Application Information
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