A strain of Providencia stutzeri jd and its application
A technology of Providencia stutzeri and Providencia spp. is applied in the field of microorganisms and can solve problems such as the inability to meet the bioremediation of pesticide residue pollution
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Embodiment 1
[0023] Embodiment 1: Isolation and identification of Providencia stutzeri JD
[0024] 1) Isolation and screening of Providencia stutzeri JD
[0025] Weigh 10.0g of soil polluted by sclerotia for a long time and place it in a 250mL Erlenmeyer flask, add 100mL of sterilized inorganic salt medium, add pesticide standard sample to 25mg / L, place on a shaker, and set the temperature at 30°C and 150rpm Under the condition of shaking culture 5d. Take 0.5mL supernatant from the culture solution and put it in a new inorganic salt culture solution containing 25mg / L pesticide, cultivate it under the same conditions until the culture solution becomes turbid, then take the supernatant and inoculate it in a new culture medium with 10% inoculation amount. Culture solution, repeat the above operations, and gradually increase the concentration of pesticides to 1000mg / L (pesticide concentrations are respectively 25mg / L, 50mg / L, 75mg / L, 100mg / L, 1000mg / L), to obtain enriched degrading bacteria l...
Embodiment 2
[0050] Embodiment 2: the preparation of degrading bacteria Providencia stutzeri JD preparation
[0051] 1) Preparation of liquid degrading bacterial agent
[0052] Including the following steps:
[0053] (1) Inoculate 5% bacterial suspension of strain JD in beef extract peptone salt medium, shake culture at 35° C., 150-200 rpm for 5-12 hours, and obtain strains.
[0054] (2) Inoculate the strain into a seed bottle at an inoculum amount of 1% of the volume ratio of the beef extract peptone salt medium, and vibrate at 35° C. for 5 to 12 hours at 150 to 200 rpm to obtain a seed liquid.
[0055] (3) Inoculate the obtained seed liquid into the fermentation medium (beef extract 7g / L, yeast extract 14g / L and sodium chloride 5g / L) according to the inoculum size of 10% volume ratio, at 35°C, 150~ Ferment and cultivate at 200rpm for 12 to 15 hours to obtain a fermented liquid (OD 600 greater than 20), the fermentation broth is directly diluted to be the JD liquid bacterial agent, and...
Embodiment 3
[0061] Embodiment 3: Providencia stutzeri JD is to the degradative efficiency of sclerotia net
[0062] Add sclerotium in the inorganic salt medium (with embodiment 1), make its final concentration be 50.0mg / L; Appropriate thalline is added in the inorganic salt medium containing sclerotium 50mg / L, adjust its OD600 to be 0.2 , cultured in a shaker at 30°C and 150rpm for 7 days, and samples were taken every 24 hours to determine the degradation ability of the strain. The non-inoculated inorganic salt medium added with the same concentration of pesticide was used as the control.
[0063] Apply gas chromatography to measure the residual amount of sclerotia net and calculate the degradation rate, the results are as follows: figure 2 As shown, the results show that the degrading bacteria JD can effectively degrade sclerotia in a short period of time, and the degradation rate of sclerotia with an initial concentration of 50 mg / L can reach 59.67% within 5 days, and 70.75% of sclero...
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