Degrading strain for dimethyltetrachloro (MCPA) of phenoxyacetic acid herbicide and microbial agent produced by degrading strain

A technology of phenoxyacetic acid and dimethyltetrachloride, which is applied in the bio-high field, can solve the problems of phytotoxicity of the next crops, and achieve the effects of low production and use costs, pollution control and good repair effect

Active Publication Date: 2020-08-28
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, due to the extensive use of the herbicide MCPA, the problem of phytotoxicity of the next crops caused by its residues has become increasingly prominent

Method used

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  • Degrading strain for dimethyltetrachloro (MCPA) of phenoxyacetic acid herbicide and microbial agent produced by degrading strain
  • Degrading strain for dimethyltetrachloro (MCPA) of phenoxyacetic acid herbicide and microbial agent produced by degrading strain
  • Degrading strain for dimethyltetrachloro (MCPA) of phenoxyacetic acid herbicide and microbial agent produced by degrading strain

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The isolation and identification of embodiment 1 bacterial strain

[0033] The invention provides a bacterial strain capable of efficiently degrading MCPA and the bacterial agent produced therefrom. The bacterial strain used is a Gram-negative bacterium DCP-6d, which is isolated from the soil of a pesticide factory in Nanjing, Jiangsu. The specific isolation and screening methods for strains are as follows:

[0034] Take 10.0g contaminated soil sample and add to 100mL containing 30mg L -1 Inorganic salt (hereinafter referred to as MM) medium of MCPA, 30 ℃, 150rpm shaker culture 7d, with 15% inoculum size (v / v) transferred to fresh medium through the same treatment, continuous enrichment culture four times . Ultraviolet spectrophotometer was used to detect the degradation of the fifth generation enrichment solution. The effective fifth-generation enrichment solution containing 30mg L -1 MCPA was diluted and coated on MM solid medium, and cultured at 30°C for 5 days. ...

Embodiment 2

[0036] Embodiment 2 laboratory degradation experiment

[0037] 2.1 Growth utilization and degradation of MCPA by strain DCP-6d

[0038] Detection of DCP-6d by high performance liquid chromatography: take 3mL sample, centrifuge at 12,000rpm for 5min, carefully absorb the supernatant, add 25% hydrochloric acid, and adjust the pH to about 3.0. Then, extract with an equal volume of dichloromethane, remove water with anhydrous sodium sulfate, take 2mL of dichloromethane and blow dry, then redissolve in 0.5mL of methanol, filter with an organic phase filter membrane with a pore size of 0.22 μm, and detect by HPLC. Detection conditions: Shimadzu RID-10A is the high-performance liquid chromatograph; the chromatographic column is a C18 reversed-phase column with a size of 250mm×4.6 mm; the column temperature is 40°C; the mobile phase is methanol:water:acetic acid (80:20:0.5, V :V:V), the flow rate is 1.0mL min L -1 ; The detection wavelength is 220nm and 235nm.

[0039] The bacteria...

Embodiment 3

[0051] Embodiment 3 bacterial agent preparation

[0052] The original seed of the halogenated phenol-degrading strain DCP-6d of the present invention is activated on a petri dish, and inoculated on the inclined surface of a test tube for later use. The test tube was inoculated in 200mL LB medium (LB medium formula: peptone 10g L -1 , yeast powder 5g L -1 , NaCl 5g L -1 , pH 7.4) in a 1,000mL shake flask, cultured with constant temperature shaking until the logarithmic phase, ready to inoculate the primary seed tank. The first-level seed tank is 50L, the feeding volume is 40L, and the medium formula is: glucose 8g L -1 , yeast extract 5g L -1 , K 2 HPO 4 1g L -1 , NaCl 5gL -1 , CaCO 3 2g L -1 , MgSO 4 0.2gL -1 , soybean oil 0.1% (v / v), pH value 7.5; after feeding, high-pressure steam sterilization, after cooling to 30 ℃, inoculate 50 L first-level For seed tanks, cultivate until the logarithmic growth phase, the stirring speed is 220rpm, and the sterile air flow...

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Abstract

The invention provides a degrading strain for dimethyltetrachloro (MCPA) of a phenoxyacetic acid herbicide and a microbial agent produced by the degrading strain, wherein the adopted strain is a gramstaining reaction negative strain DCP-6d, the strain is identified as Pigmentiphaga sp., the strain is deposited in the China Center for Type Culture Collection on December 23, 2019, and the preservation number of the strain is CCTCC M 20191085. The degrading strain DCP-6d for the MCPA of the phenoxyacetic acid herbicide can be applied to degradation of the MCPA and 2,4-dichlorphenoxyacetic acid (2,4-D), and is preferentially applied to degradation of the MCPA in soil; and a degrading microbial agent product is directly applied so that the residual quantity of the MCPA in the soil can be decreased by 90% or above, and the pollution problem of the MCPA in the soil is solved.

Description

technical field [0001] The invention belongs to the high-tech field of biology, and relates to a degrading bacterial strain of the phenoxyacetic acid herbicide dimethyltetrachloride and a bacterial agent produced therefrom, which mainly uses microorganisms to efficiently remove the phenoxyacetic acid herbicides MCPA and 2 in the environment. 4-dichlorophenoxyacetic acid (2,4-D) pollution, suitable for microbial enhanced remediation of soil and water pollution. [0002] technical background [0003] The use of chemical pesticides has greatly promoted the development of modern agriculture, saved a lot of losses caused by diseases, insect pests and weeds for agricultural production, and greatly improved the labor efficiency and mechanization degree of agricultural production. In the process of modernization of agricultural production played an immeasurable role. But at the same time, pesticides are some toxic and harmful chemicals. After they enter the ecological environment, t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20B09C1/10C02F3/34C12R1/01C02F101/36
CPCC12N1/20B09C1/10C02F3/34C02F2101/36C12R2001/01C12N1/205Y02A50/20
Inventor 蒋建东张龙徐希辉
Owner NANJING AGRICULTURAL UNIVERSITY
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