Pseudomonas stutzeri JSD-008 and its degradation function for organophosphorus pesticide

A technology of Pseudomonas stutzeri and organophosphorus pesticides, applied in the field of chemical pesticide biodegradation, can solve the problems of chlorpyrifos degradation and discovery of related functions that have not been reported

Inactive Publication Date: 2008-01-02
谢明
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

In terms of patents, it is mainly about the strains and applications of plant disease control, aquatic disease control, sanitary descaling, petrochemical desulfurizat

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  • Pseudomonas stutzeri JSD-008 and its degradation function for organophosphorus pesticide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0117] Embodiment 1: bacterial strain screening method

[0118] Acclimatization and cultivation of strains: take 5g of soil samples from pesticide-contaminated areas, inoculate them into a 50ml triangular flask with chlorpyrifos as the only carbon source or the only nitrogen source selective medium under aseptic conditions, add the original drug of chlorpyrifos, Make the final concentration 300mg / L, place in 30 ℃, 180rpm shaker culture, transfer according to 10% inoculum amount every 7 days, increase the concentration of chlorpyrifos each time by 200mg / L, until the chlorpyrifos in the culture medium The final concentration reaches 1100mg / L.

[0119] Isolation and purification of bacterial strains: Take the culture after acclimatization and culture, and serially dilute 10 -6 , 10 -7 , 10 -8 Spread on the plate of the selective medium (inorganic salt medium with chlorpyrifos as the only carbon source or inorganic salt medium with chlorpyrifos as the only nitrogen source) used...

Embodiment 2

[0120] Embodiment 2: bacterial strain is to the degradation efficiency of chlorpyrifos

[0121] Inoculate the isolated and purified bacterial strains on the selective medium used for isolation. After they grow vigorously, make a bacterial suspension and inoculate them in 50ml of inorganic salt medium containing 50mg / L chlorpyrifos. At the same time, set a blank for no inoculation As a control, cultured on a shaker (30° C., 180 rpm) for 5 days, extracted with dichloromethane, measured the content of chlorpyrifos in the culture solution by gas chromatography, and calculated the degradation rate of the strain to chlorpyrifos.

[0122] Degradation rate% = (residual pesticide amount in the system after cultivation / initial pesticide addition amount) × 100%

[0123] The results showed that the degradation rate of JSD008 strain to chlorpyrifos reached 85.7%, the degradation rate of the blank control cultured with chlorpyrifos as the only carbon source of inorganic salt was 15.6%, and ...

Embodiment 3

[0124] Embodiment 3: the 16S rDNA sequence determination of bacterial strain

[0125] Extraction of total DNA: Take 1.5ml of bacterial solution cultured overnight, centrifuge at 5000rpm for 2min, collect the precipitate; add 600ul 65℃ preheated cell lysate, mix well, add 1 / 10 volume of Tris saturated phenol, and keep warm at 65℃ Half an hour, and shake gently from time to time; add an equal volume of phenol / chloroform / isoamyl alcohol, mix well, centrifuge at 13000rpm, 10min; take the supernatant, add an equal volume of chloroform / isoamylalcohol and extract again, centrifuge, 13000rpm, 10min ; Take the supernatant, add 1 / 10 volume of 3M sodium acetate, add 0.6 volume of pre-cooled isopropanol, and place it at -70°C for 10 minutes; centrifuge at 10,000 rpm for 10 minutes, discard the supernatant, wash the precipitate with 70% ethanol, and dry it in vacuum; Add 200ul of deionized water to redissolve, add 20ul of 1mg / ul RNase, heat at 65°C for 30min; add water to 500ul, add an equ...

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Abstract

The invention discloses a Pseudomonas stutzeri JSD-008 and degradation for organophosphorus pesticide, which is preserved in the 'Chinese germ management committee center (CGMCC)' with preservation number at CGMCC No.1738, wherein the strain is separated from the soil at pollution discharge pore of pesticide processing plant polluted by organophosphorus pesticide; the thallus and ferment liquid with the thallus can be degradation agent of organophosphorus pesticide to degrade the chlorpyrifos into trichlopyridinenol and methyl parathion into nitrophenol; fitting for rapid in-situ rehabilitation polluted by field soil pesticide.

Description

technical field [0001] The invention belongs to the technical field of chemical pesticide biodegradation, and relates to a strain of Pseudomonas stutzeri JSD-008 and its degradation effect on organophosphorus pesticides chlorpyrifos and methyl parathion, which can be used for in-situ restoration of soil pesticide pollution project. Background technique [0002] Chemical pesticides are important means of agricultural production and play an important role in the high yield and harvest of crops. Some of them have stable structures and are not easy to degrade, which often cause serious environmental pollution. Scientific research has found that some specific microbial groups in nature have the function of decomposing and transforming pesticides, and can decompose chemical pesticides into non-toxic compounds. This process is called biodegradation. However, the number of such microorganisms in nature is limited. In order to speed up this process, it can be achieved by adding micr...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N15/31A62D3/00C12R1/38
Inventor 谢明万方浩周淑云邱卫亮
Owner 谢明
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