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Aryloxyphenoxypropionate weed killer degradation strain and application thereof

An aryloxyphenoxypropionic acid and herbicide technology, which can be applied to bacteria, isolated microorganisms, microorganisms, etc., can solve the problems of narrow degradation spectrum, weak degradation ability of AOPP herbicide, ecological pollution, etc., and achieve the effect of rich degradation spectrum.

Active Publication Date: 2017-03-08
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a kind of high-efficiency broad-spectrum degrading microorganisms for AOPP herbicides in view of the ecological pollution caused by AOPP herbicide residues, and the existing microorganisms have weak degradation ability and narrow degradation spectrum to AOPP herbicides

Method used

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  • Aryloxyphenoxypropionate weed killer degradation strain and application thereof
  • Aryloxyphenoxypropionate weed killer degradation strain and application thereof
  • Aryloxyphenoxypropionate weed killer degradation strain and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0042] Aryloxyphenoxypropionic acid herbicide degrading strain Acinetobacter sp. Isolation and screening of VL-2:

[0043] Take 2 g of the sludge sample from the sewage discharge outlet of the pesticide factory and place it in 100 mL of inorganic salt medium containing 20 mg / L fenoxaprop-ethyl, at 30 °C, 180 r min -1 Cultured for 5 days. Use the ultraviolet scanner to measure the degradation situation of the enriched liquid to fenoxaprop-p-ethyl, and after confirming that fenoxaprop-p-ethyl is degraded, the culture solution in the inorganic salt medium is inserted into the 50mg / L of fenoxaprop-p-ethyl in the inorganic salt medium, continue to enrich and measure the degradation situation, follow this method until the concentration of fenoxaprop-p-p-ethyl is increased to 100 mg / L, and pass passage 3 times.

[0044] The enrichment solution with the degradation performance of fenoxaprop-p-p-p was serially diluted and spread on the inorganic salt medium plate with 100 mg / L oxa...

Embodiment 2

[0048] Aryloxyphenoxypropionic acid herbicide degrading strain Acinetobacter sp. Identification of VL-2 and its growth characteristics:

[0049] 16S rDNA identification of strain VL-2:

[0050] Using primers 27F: 5`-AGAGTTTGATCCTGGCTCAG-3` and 1492R: 5`-TACCTTGTTACGACTT -3` to amplify the 16S rDNA of strain VL-2, connect it to the cloning vector pMD19T by T / A cloning, and construct the recombinant cloning vector pMD19T- 16S, which was transformed into the cloning host strain Escherich coli DH5α obtained from recombinant microorganisms Escherich coli DH5α (pMD19T-16S), the obtained exogenous fragment of the recombinant microorganism was sequenced, and the 16S rDNA sequence was compared with the NCBI database, and the strain VL-2 was identified as Acinetobacter at the molecular level.

[0051] Growth characteristics of strain VL-2:

[0052] Acinetobacter sp. VL-2 is a Gram-negative bacterium that grows very quickly on LB plates. At 30°C for 12 hours, it can form sli...

Embodiment 3

[0054] This example illustrates the optimal growth conditions for strain VL-2.

[0055] Will Acinetobacter sp. VL-2 colonies were inoculated into liquid LB medium at 20, 25, 30, 37, 42°C; pH 4, 5, 6, 7, 8, 9, 10, 11; salt concentration 0 g L -1 , 2 g L -1 , 4 g L -1 , 6 g L -1 , 8 g L -1 , 10 g L -1 , 12 g L -1 , 16 g L -1 , 20 g L -1 ;Measure the OD600 value after shaking culture for 12 hours under the conditions of 10%, 20%, 30%, 40%, 50%, 60%, and 70% of the medium loading volume to determine the optimum fermentation conditions.

[0056] Wherein, the LB medium formula is: peptone 10.0 g·L -1 , yeast powder 5.0 g·L -1 , NaCl10.0 g·L -1 , pH 7.0, sterilized at 121°C for 20 min.

[0057] The optimum growth temperature of strain VL-2 is 30°C, and it can grow well at 25-42°C, but its growth is slightly inhibited at lower temperatures (20°C and 25°C). Bacterial strain VL-2 grows well when the pH is between 5-11, and its optimum growth pH is 7; The growth decreased ...

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Abstract

The invention relates to an aryloxyphenoxypropionate weed killer degradation strainand an application thereof. The strain is named as Acinetobacter sp. VL-2 and was preserved in the China Center for Type Culture Collection (CCTCC) on July, 4th, 2016 with the preservation number of CCTCC NO: M 2016374, and the nucleotide sequence of 16S r DNA is indicated as SEQ ID NO: 1 in a sequence table. The invention further provides the application of the strain in AOPP weed killer degradation. The strain VL-2 can completely degrade 100mg / L of fenoxaprop-p-ethyl in 120 hours, and being served as only carbon source growth, the strain VL-2 can degrade other AOPP weed killers of cyhalofop-butyl, clodinafop-propargyl, quizalofop-p-thyl, efficient haloxyfop-R-methyl and the like. Important application value on biological treatment of AOPP weed killer contaminated soil is achieved.

Description

technical field [0001] The invention belongs to the technical field of biological treatment, and relates to an aryloxyphenoxypropionic acid herbicide-degrading bacterial strain and application thereof. Background technique [0002] Aryloxyphenoxypropionic acid (AOPP) herbicides are mainly used to control grass weeds, and are widely used in crop fields in my country and the world because of their high efficiency and low toxicity. Fenoxaprop-ethyl is the most widely used herbicide in this category, with sales of US$ 250 million and US$ 280 million in 2003 and 2009, accounting for 31.2% of the total sales of this herbicide. The seventh place in the sales list of herbicides. During the application process, only a small part of AOPP herbicides can attach to plants and play a role, and most of the rest remain in the ecological environment such as air, soil and water. Although the toxicity of AOPP herbicides is relatively low, a large number of studies have shown that AOPP herbic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02B09C1/10C12R1/01
CPCC12N1/02B09C1/10C12R2001/01C12N1/205
Inventor 董维亮史周昆马江锋章文明姜岷
Owner NANJING UNIV OF TECH
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