Screening method for degradation bacteria with polycyclic aromatic hydrocarbon as substrate

A polycyclic aromatic hydrocarbon and screening method technology, applied in the field of contaminated soil bioremediation, can solve the problems of large workload, time-consuming and laborious, and high cost, and achieve the effects of saving time, shortening the ascending distance, and reducing waste

Inactive Publication Date: 2012-07-04
SHENYANG INST OF APPL ECOLOGY CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The screening of traditional PAHs-degrading functional bacteria often has a certain degree of blindness and randomness, and the workload is heavy, time-consuming and labor-intensive, and the cost is high

Method used

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  • Screening method for degradation bacteria with polycyclic aromatic hydrocarbon as substrate
  • Screening method for degradation bacteria with polycyclic aromatic hydrocarbon as substrate
  • Screening method for degradation bacteria with polycyclic aromatic hydrocarbon as substrate

Examples

Experimental program
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Effect test

Embodiment 1

[0026] 1) Collect the surface 0-20cm soil of the Shenfu irrigation area as a sample, and store it in a refrigerator at 4°C. Weigh 5g of the soil sample for testing and insert it into 45ml of liquid inorganic salt culture medium containing tetracyclic polycyclic aromatic hydrocarbon pyrene, wherein the concentration of pyrene is 100㎎ L -1 , add an appropriate amount of glass beads, and place on a constant temperature shaker at 28-32°C at 180 r·min -1 Enrichment culture for 1 month, after enrichment culture, pipette the first culture solution into 45ml liquid inorganic salt medium containing pyrene at a volume ratio of 10%, carry out second enrichment culture, repeat 4 times in total , every transfer, the concentration of pyrene increased by 100㎎·L -1 , that is, the last time the concentration of pyrene was increased to 400㎎·L -1 .

[0027] 2) Under sterile conditions, the enriched culture solution was serially diluted 10 times with sterile water to 10 -7 , take 10 respectiv...

Embodiment 2

[0040] 1) Collect the surface 0-20cm soil of the Shenfu irrigation area as a sample, and store it in a refrigerator at 4°C. Weigh 5g of the soil sample to be tested, and insert it into 45ml of liquid inorganic salt medium containing tricyclic polycyclic aromatic hydrocarbon phenanthrene (acenaphthene, fluorene, anthracene, fluoranthene, etc.), where the concentration of phenanthrene is 200㎎ L -1 , add an appropriate amount of glass beads, and place on a constant temperature shaker at 28-32°C for 180r·min -1 Enrichment culture for 1 month, after enrichment culture, pipette the first culture solution into 45ml liquid inorganic salt medium containing phenanthrene at a volume ratio of 10%, carry out second enrichment culture, repeat 4 times in total , each transfer, the concentration of phenanthrene increased by 100㎎·L -1 , that is, the last time the concentration of phenanthrene was increased to 500㎎·L -1 .

[0041] 2) Under sterile conditions, the enriched culture solution wa...

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Abstract

The invention relates to the technical field of biological remediation of polluted soil, in particular to a screening method for degradation bacteria with polycyclic aromatic hydrocarbon as a substrate. The method comprises the following steps of: performing enrichment culture on the degrading bacteria with the polycyclic aromatic hydrocarbon as the substrate by using a method for gradually increasing the concentration of the polycyclic aromatic hydrocarbon; after diluting enrichment culture solution by 101-107 times, coating the solution with proper concentration on a sorbitol solid culture medium; after additionally forming a pyrene film, placing in a constant-temperature culture box of 28-32 DEG C to culture for 1-3 weeks; picking up single colonies with degrading rings to be dibbled in a beef extract peptone culture medium containing polycyclic aromatic hydrocarbon respectively; placing in the constant-temperature culture box of 28-32 DEG C to culture for 1-2 weeks; dibbling grown strains in an inorganic salt solid culture medium with a polycyclic aromatic hydrocarbon membrane; and placing in the constant-temperature culture box of 28-32 DEG C to culture for 1-3 weeks so as to grow the strain, namely the degrading strain with the polycyclic aromatic hydrocarbon as a unique carbon source. By using the method, the polycyclic aromatic hydrocarbon degrading bacteria can be quickly screened in a laboratory; and a quick and convenient means is provided for developing a new biological remediation microbial resource.

Description

technical field [0001] The invention relates to the technical field of contaminated soil bioremediation, in particular to a screening method for degrading bacteria using polycyclic aromatic hydrocarbons as substrates. Background technique [0002] Polycyclic aromatic hydrocarbons (PAHs) refer to a class of unique persistent organic pollutants composed of two or more benzene rings in the form of linear, angular or clustered. Many PAHs will accumulate in organisms and cause carcinogenic, teratogenic, and mutagenic effects on organisms through cytotoxicity, genotoxicity, and immunotoxicity, posing a huge threat to natural biological safety and human health. In the 1980s, the U.S. Environmental Protection Agency included 16 PAHs without branches in the list of priority pollutants, and my country also included PAHs in the blacklist of environmental pollutants. Therefore, remediation of PAHs-contaminated soil is a very urgent task. [0003] Microbial degradation of PAHs has the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04C12N1/26
Inventor 苏振成胡凤钗李新宇王秀娟张惠文
Owner SHENYANG INST OF APPL ECOLOGY CHINESE ACAD OF SCI
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