Biological restoration method for p-chloronitrobenzene compound polluted environment
A technology of chloronitrobenzene and p-chloronitrobenzene, which is applied in the restoration of polluted soil, chemical instruments and methods, biological water/sewage treatment, etc. Effect
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Embodiment 1
[0029] Embodiment 1, detect the repair ability of the inventive method to the soilless substrate polluted by p-chloronitrobenzene compounds
[0030] Taking p-chloronitrobenzene as an example, the method for detecting the repair ability of the soilless substrate polluted by p-chloronitrobenzene compounds, the specific process comprises the following steps:
[0031] 1) Inoculate Comamonas testosteroni (Comamonas testosteroni) CNB-1 CGMCC No.1028 in an inorganic salt medium with p-chloronitrobenzene as a substrate, and vibrate at 28°C and 130rpm for 16 hours to obtain seeds liquid;
[0032] 2) Inoculate the seed solution of Comamonas testosteroni CNB-1 CGMCC No.1028 obtained in step 1) into LB liquid medium at an inoculation ratio of 1%, and culture it with shaking at 30° C. for 18 hours;
[0033] 3) Collect the bacteria by centrifugation at 6000rpm, and resuspend it in a sterile solution with a mass percent concentration of 0.9% NaCl to make the OD 600 Reach 2.0, obtain the ba...
Embodiment 2
[0037] Embodiment 2, detection method of the present invention is to the remediation ability of the soil polluted by p-chloronitrobenzene compounds
[0038] Taking p-chloronitrobenzene as an example, the pot experiment is used to detect the repair ability of the inventive method to the soil polluted by p-chloronitrobenzene compounds, and the specific process comprises the following steps:
[0039] 1) Inoculate Comamonas testosteroni (Comamonas testosteroni) CNB-1 CGMCC No.1028 in an inorganic salt medium with p-chloronitrobenzene as a substrate, and vibrate at 30°C and 130rpm for 16 hours to obtain seeds liquid;
[0040] 2) Inoculate the seed solution of Comamonas testosteroni CNB-1 CGMCC No.1028 obtained in step 1) into LB liquid medium at an inoculation ratio of 1%, and culture it with shaking at 28° C. for 18 hours;
[0041] 3) Collect the bacteria by centrifugation at 7000rpm, and resuspend it in a sterile solution with a mass percent concentration of 0.9% NaCl to make th...
Embodiment 3
[0046] Embodiment 3, Comamonas testosteroni (Comamonas testosteroni) CNB-1 CGMCC No.1028 in the detection of alfalfa root colonization ability
[0047] In order to facilitate the observation of the colonization of p-chloronitrobenzene-degrading strain Comamonas testosteroni (Comamonas testosteroni) CNB-1 CGMCC No.1028 in the plant root system during the bioremediation process, green fluorescent protein (green fluorescence protein, GFP ) gene is integrated on the genomic DNA of Comamonas testosteroni (Comamonas testosteroni) CNB-1, and specific method is: (1) preparation of Comamonas testosterone (Comamonas testosteroni) CNB-1 competent cell: summary of the invention The seed solution obtained in step 1) was inserted into LB liquid medium according to the inoculation amount of 1%, and cultured with shaking at 130rpm at 28°C to make its OD 600reach 0.3-0.4, (the following operations are all carried out at 4°C) collect the bacterial cells by centrifugation at 4500rpm, resuspend t...
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