A Strain of Stenotrophomonas and Its Application on Codonopsis Codonopsis
A technology of Stenotrophomonas and Codonopsis pilosula, applied in the field of microorganisms, can solve the problems of discounted effect, long half-life, damage, etc., and achieve the effects of improving quality safety, good ecological safety, and solving pollution problems
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Embodiment 1
[0032] Example 1 Isolation of Stenotrophomonas ZBD-a-03-I CGMCC No.17102
[0033] 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 an...
Embodiment 2
[0034] Example 2 Identification of Stenotrophomonas ZBD-a-03-I CGMCC No.17102.
[0035] The isolated and purified ZBD-a-03-I strain was identified from the following three aspects.
[0036] 1. Morphological identification
[0037]Cultivate the ZBD-a-03-I isolated in Example 1 and observe the colony, mainly including the size, color, transparency, humidity, and surface state of the colony (whether it is flat, convex, wrinkled, depressed, etc.) , Colony edge state (whether neat, irregular, radial, etc.). Microscopic examination was performed after staining with biological stains.
[0038] The results showed that the colony of strain ZBD-a-03-I obtained after separation and purification: the diameter of the colony was 6-7 mm, the color of the colony was light yellow, the colony was opaque, the surface of the colony was moist and slightly raised, and the edges of the colony were neat. Microscopic examination results were rod-shaped, Gram-negative, with flagella and no spores. ...
Embodiment 3
[0047] Example 3 Nitrogenase activity test of Stenotrophomonas ZBD-a-03-I CGMCC No.17102
[0048] The Stenotrophomonas ZBD-a-03-I CGMCC No.17102 obtained by the separation and purification of Example 1 was tested for nitrogenase activity, and the acetylene reduction method was used. The specific method is as follows: ZBD-a-03-I was prepared and obtained Strain suspension (cells, 1 x 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...
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