Sphingomonas sp. and its application
A technology of sphingomonas and bacterial agent, which is applied in the field of microorganisms, can solve the problems of excessive pesticide residues, chlorpyrifos residual ecological environment, and export impact, and achieve the effects of enhancing activity, being suitable for widespread use, and efficiently degrading pesticide residues
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Embodiment 1
[0030] Example 1 screening strains
[0031] 1. Obtaining the strain
[0032] (1) Sample collection: In October 2012, fresh plant samples of chives domesticated by chlorpyrifos selective pressure were collected in the experimental field of Jiangsu Academy of Agricultural Sciences in Nanjing City, Jiangsu Province. The dust and soil attached to the plant surface were washed with tap water, dried naturally, and pressed. Surface disinfection according to the above procedure: soak in 75% alcohol for 3-5 minutes, then rinse with sterile water for 3-4 times, and then rinse with 2.5% NaClO 2 Rinse for 2-5 minutes, and finally wash 5 times with sterile water;
[0033] (2) Isolation and screening of bacterial strains: Grind the roots, stems and leaves of Chinese chives respectively, absorb the juice and spread it evenly on the enriched medium plate, and culture at 30° C. in the dark for 2-7 days.
[0034] (3) Strain enrichment culture: After the colony grows, pick a single colony an...
Embodiment 2
[0042] Embodiment 2 prepares HJY bacterial agent
[0043] 1. Strain activation: Pick the Sphingomonas HJY obtained in Example 1 to the strain preservation medium, streak and pick a single colony twice at 30°C (24 hours each), and then pick A single colony is cultured in the strain activation medium at 30°C, 150-200rpm shaker for 12-24h;
[0044] 2. Preparation of liquid seeds: inoculate the bacteria species activated in step 1 according to the inoculum volume of 1% of the volume of the seed medium in the fermenter equipped with the seed medium, and cultivate in air at 25-38°C for 16-24h to obtain liquid seeds;
[0045] 3. Liquid fermentation: Inoculate liquid seeds into a fermenter equipped with seed medium, the inoculation amount is 1% of the volume of the medium, and culture at 30-35°C and 150rpm in the dark for 4-6 hours to obtain live bacteria Cultures;
[0046] 4. Bacterial agent preparation: Take 50ml of live bacterial culture and centrifuge at 4°C and 5000rpm for 1...
Embodiment 3
[0047] Example 3 HJY bacterial agent degrades chlorpyrifos under in vitro conditions
[0048] 1. Inoculate 2ml of the HJY bacterial agent obtained in Example 2 into 100ml of MSM medium, add chlorpyrifos to a final concentration of 20mg / L, and measure the initial OD of the medium 600 and the content of chlorpyrifos; at the same time, the culture medium not inoculated with HJY bacterial agent (adding the same volume of sterile saline as the bacterial agent) was used as the control group, and each treatment was repeated 3 times;
[0049] Both the control group and the experimental group were cultured at 30°C and 200rpm in the dark, and samples were taken at different times to determine the OD 600 value and chlorpyrifos residue; after 12 days, sampling and extraction were carried out, and the content of chlorpyrifos o-phenyl was measured by HPLC, and the measurement results were as follows: Figure 4 As shown, in the MSM medium with 20mg / L chlorpyrifos as the single carbon sour...
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