Strain for degrading phenanthrene and application thereof in soil remediation
A technology for bacterial strains and polluted soil, applied in the field of environmental microbiology, can solve the problem of long degradation cycle of bacterial strains, and achieve excellent degradation effect, fast and efficient degradation effect.
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Embodiment 1
[0031] Embodiment 1 Enrichment culture, purification, screening and identification of bacterial strains of the present invention
[0032] (1) culture medium
[0033] (I) MSM medium: NaN03 4.0g, NH4Cl2 2.0g, KH2P04 1.5g, Na2HP040.5g, CaCl20.01g, MgCl2 0.2g, 1ml trace element solution, deionized water 1L, adjusted to pH=7.0, extinguished at 121°C Bacteria 20min. (A certain amount of phenanthrene is added during use, and since phenanthrene is semi-volatile, it is necessary to add phenanthrene on the ultra-clean workbench after the medium is sterilized.)
[0034](II) Selection medium (solid medium): NaN03 4.0g, KH2P04 1.5g, Na2HP04 0.5g, CaCl20.01g, MgCl2 0.2g, agar 20.0g, trace element solution 1ml, deionized water 1L, pH=7.0, Sterilize at 121°C for 20 minutes.
[0035] (III) Enrichment medium: 10.0 g of peptone, 5.0 g of yeast extract, and 10.0 g of NaCl were dissolved in 1 L of deionized water, adjusted to pH = 7.0 with NaOH and HCl, and sterilized at 121°C for 20 minutes. ...
Embodiment 2
[0049] The cultivation of embodiment 2 strain Pseudomonas sp.Lphe-2
[0050] (1) The culture condition optimization of bacterial strain of the present invention
[0051] Seed solution preparation: take the preserved slant and place it for 2 hours at room temperature, use a sterile dropper to draw 0.5ml of inorganic salt medium (the composition of the inorganic salt medium: NaNO 3 4.0g, NH 4 Cl 2 2.0g, KH 2 PO 4 1.5g, Na 2 HPO 4 0.5g, CaCl 2 0.01g, MgCl 2 0.2g, 1ml trace element solution, 1L deionized water, adjust pH=7.0, sterilize at 121°C for 20min; the composition of the trace element solution: FeSO 4 ·7H 2 O 2g, ZnSO 4 ·7H 2 O 0.03g, H 3 BO 3 0.3g, CoCl 3 ·6H 2 O 0.3g, NiCl 2 ·6H 2 O0.02g, Na 2 MoO 4 2H 2 O 0.03g, deionized water 1L, adjust pH=7.0. ) was dropped onto the slope, and after slight shaking, 0.2ml of the bacterial suspension was streaked on the solid medium and cultured for 96 hours. The strains with good growth conditions were picke...
Embodiment 3
[0060] Application of Example 3 bacterial strain Pseudomonas sp.Lphe-2 in remediation of phenanthrene and pyrene contaminated soil
[0061] The soil samples used in this experiment were collected in the garden on the south side of the west gate of the School of Environmental Measurement, China University of Mining and Technology. The five-point method was used to sample after removing the surface vegetation. After the collected soil samples were naturally air-dried, impurities such as stones and branches were removed, and then passed through a 2mm sample sieve. Crush the clods > 2mm and pass through a 2mm sample sieve, mix well and set aside.
[0062] The air-dried and sieved soil samples were sent to the Red Soil Station of the Chinese Academy of Sciences for the determination of physical and chemical properties. Specifically, it includes six indicators: pH, organic carbon, total nitrogen, total phosphorus, available phosphorus and available potassium. The mud preparation met...
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