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A screening method and application of yew rhizosphere PAH-degrading bacteria

A polycyclic aromatic hydrocarbon and yew technology, applied in the field of microorganisms, can solve problems such as polycyclic aromatic hydrocarbon pollution, and achieve the effects of no secondary pollution and low repair cost.

Inactive Publication Date: 2018-06-19
DALIAN JIAOTONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method domesticates the rhizosphere fungi of Taxus chinensis, and screens out three strains that use pyrene as the sole carbon source to solve the pollution problem of polycyclic aromatic hydrocarbons in the environment

Method used

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  • A screening method and application of yew rhizosphere PAH-degrading bacteria
  • A screening method and application of yew rhizosphere PAH-degrading bacteria
  • A screening method and application of yew rhizosphere PAH-degrading bacteria

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1 product liquid phase experiment

[0036] 1.1 Experimental reagents

[0037] Benzopyrene (97%), CAS No.: 50-32-8, Bailingwei Technology Co., Ltd.; Pyrene (97%), CAS No.: 129-00-0, Aladdin Reagent Co., Ltd.; Phenanthrene (97%), CAS No.: 85-01-8, Aladdin Company; Naphthalene (95%), CAS No.: 91-20-3, Shanghai Xinyu Biotechnology Co., Ltd.; Acenaphthene (95%), CAS No.: 83-32-9, Shanghai Xinyu.

[0038] 1.2 Medium and solution

[0039] (1) Polycyclic aromatic hydrocarbons in acetone solution: Dissolve 0.25 g of polycyclic aromatic hydrocarbons in 50 mL of acetone solution to prepare a 5 g / L stock solution for future use.

[0040] (2) Seed medium: beef extract 5g, peptone 10g, NaCl 5g, distilled water 1000mL.

[0041] (3) Inorganic salt medium: phosphate buffer solution 6mL, MgSO 4 ·H 2 O (24.5g / L) 3mL, CaCl 2 (1g / L)1mL, FeCl 3 (1g / L)2mL, ZnSO 4 ·H 2 O(1g / L)2mL, MnSO 4 ·H 2 O (0.5g / L) 1mL, dilute to 1L.

[0042] (4) Phosphate buffer solution: K 2 HPO...

Embodiment 2

[0066] Example 2 Domestication, screening and identification of PAHs degrading bacteria

[0067] 2.1 Experimental materials and methods

[0068] The strains used in this experiment came from the soil depth of 10-15cm in the rhizosphere soil of Taxus chinensis.

[0069] Table 3 Experimental reagents and materials

[0070]

[0071]

[0072] 2.2 Strain screening

[0073] 2.2.1 Medium and solution

[0074] (1) Pyrene-acetone solution: Dissolve 0.25 g of pyrene in 50 mL of acetone solution to make a 5 g / L stock solution for later use.

[0075] (2) Phosphate buffer solution: K 2 HPO 4 ·3H 2 O 26.12g, KH 2 PO 4 ·H 2 O 6.8g, NH 4 Cl 5.0g, Na 2 HPO 4 12H 2 O35.8g, distilled water 1000mL.

[0076] (3) Inorganic salt medium: Take 6 mL of the above-mentioned phosphate buffer, MgSO 4 ·H 2 O (24.5g / L) 3mL, CaCl 2 (1g / L)1mL, FeCl 3 (1g / L)2mL, ZnSO 4 ·H 2 O(1g / L)2mL, MnSO 4 ·H 2 O (0.5g / L) 1mL, dilute to 1L.

[0077] (4) Seed medium: beef extract 5g, peptone 10g, ...

Embodiment 3

[0104] Example 3 Degradation of domesticated bacteria to pyrene, phenanthrene, naphthalene, acenaphthene

[0105] 3.1 Experimental materials and methods

[0106] 3.1.1 Experimental instruments and equipment

[0107] Table 4 Instruments and Equipment

[0108]

[0109] 3.1.2 Main reagents and materials

[0110] Table 5 Experimental Reagents

[0111]

[0112]

[0113] 3.1.3 Preparation of medium and solution

[0114] (1) PAHs acetone solution: Dissolve 0.25g PAHs in 50mL acetone solution to prepare a 5g / L stock solution for later use.

[0115] (2) Seed medium: beef extract 5g, peptone 10g, NaCl 5g, distilled water 1000mL.

[0116] (3) Phosphate buffer solution: K 2 HPO 4 ·3H 2 O 26.12g, KH 2 PO 4 ·H 2 O 6.8g, NH 4 Cl 5g, Na 2 HPO 4 12H 2 O35.8g, distilled water 1000mL.

[0117] (4) Inorganic salt medium: Take 6ml of the above-mentioned phosphate buffer, MgSO 4 ·H 2 O(24.5 g / L) 3ml, CaCl 2 (1g / L)1ml, FeCl 3 (1g / L)2ml, ZnSO 4 ·H 2 O(1g / L)2ml, MnSO ...

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Abstract

The invention discloses a screening method and application of polycyclic aromatic hydrocarbon-degrading bacteria in the rhizosphere of Taxus chinensis. Take 2mL of 100g / L activated soil sample and add it to 48mL of 25mg / L pyrene solution, and vibrate at 28°C for 5 days; take 2mL of enrichment solution Added to 48mL 50mg / L pyrene solution, cultured with shaking at 28°C for 5 days; repeated cycles until the pyrene concentration was 0.2g / L. Three strains of fungi that can efficiently degrade polycyclic aromatic hydrocarbons were obtained after screening and domestication: Talaromyces, Residuum and Fusarium oxysporum. Under optimized conditions, Talaromyces can efficiently degrade a variety of polycyclic aromatic hydrocarbons, especially the degradation effect on pyrene is better than other strains. The degradation of various PAHs by Talaromyces was confirmed by HPLC and mass spectrometry. The bioflocculants extracted from Talaromyces and Residuum can also degrade polycyclic aromatic hydrocarbons such as pyrene, phenanthrene, naphthalene, and acenaphthene. The flocculation rate of the screened Talaromyces to kaolin reached 90%.

Description

technical field [0001] The invention belongs to the technical field of microbes, and relates to a screening method and application of yew rhizosphere polycyclic aromatic hydrocarbon-degrading bacteria. Background technique [0002] Polycyclic aromatic hydrocarbons (PAHs) are a class of organic compounds containing two or more benzene rings in their molecules. Such substances have stable chemical properties, high toxicity and side effects, and are likely to cause teratogenicity, carcinogenicity, and mutagenicity to humans. The deposition of polycyclic aromatic hydrocarbons in the environment mainly comes from the combustion of coal and petroleum. Industrial production, garbage incineration, and vehicle exhaust emissions will also cause a large increase in PAHs. The quantity of PAHs in the environment is widely distributed, and it is difficult to control them centrally. It is difficult to completely eliminate them with physical and chemical repair methods, and it will also ca...

Claims

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Application Information

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IPC IPC(8): C12N1/36C12R1/77C12R1/645A62D101/20C12N1/14A62D3/02
Inventor 郝大程胡文丽宋思梦朱秀华
Owner DALIAN JIAOTONG UNIVERSITY