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A degrading bacterial strain capable of simultaneously degrading two isomers of the chiral herbicide 2,4-dipropionic acid and the bacterial agent produced thereof

A herbicide and isomer technology, which is applied in the field of degrading strains and the inoculants produced by them, can solve problems such as non-target biological toxicity, and achieve the effects of increasing added value, good removal effect and protecting ecological environment.

Active Publication Date: 2022-02-25
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, the (S)-enantiomer, although not herbicidally active, may be more toxic to non-target organisms

Method used

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  • A degrading bacterial strain capable of simultaneously degrading two isomers of the chiral herbicide 2,4-dipropionic acid and the bacterial agent produced thereof
  • A degrading bacterial strain capable of simultaneously degrading two isomers of the chiral herbicide 2,4-dipropionic acid and the bacterial agent produced thereof
  • A degrading bacterial strain capable of simultaneously degrading two isomers of the chiral herbicide 2,4-dipropionic acid and the bacterial agent produced thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1, isolation and identification of bacterial strain

[0038] The invention provides a bacterial strain capable of efficiently degrading two chiral isomers of the chiral herbicide (R, S)-DCPP and the bacterial agent produced therefrom. The bacterial strain used is a Gram-negative bacterial strain DCP-6, isolated from In the soil of an abandoned pesticide factory in Nanjing, Jiangsu.

[0039] The specific isolation and screening methods for strains are as follows:

[0040] Take 10.0 g of soil sample and add it to 100 mL of liquid inorganic salt medium (hereinafter referred to as MSM) containing 30 mg / L (R,S)-DCPP, culture it on a shaker at 30 °C and 180 rpm for 7 days, and inoculate at 15% (v / v) Transplanted into fresh same medium, enriched and subcultured four times. Use an ultraviolet spectrophotometer to scan in the range of 200-350nm to detect the degradation effect of the fifth-generation enrichment solution. Dilute and spread the effective enrichment s...

Embodiment 2

[0042] Embodiment 2, laboratory degradation experiment

[0043] 2.1 Preparation of seed solution

[0044] Insert strain DCP-6 into 100mL LB medium, cultivate on a shaker at 30°C and 180rpm, collect the bacteria by centrifugation at 6,000rpm after 48 hours, wash the bacteria twice with sterilized MSM, and finally resuspend with 10mL of sterilized MSM , as a seed solution for later use.

[0045] 2.2 Degradation of chiral herbicide (R,S)-DCPP by strain DCP-6

[0046] Strain DCP-6 was inoculated into 100mL MSM containing 30mg / L (R,S)-DCPP at 5% inoculation amount, cultured on a shaker at 30°C and 180rpm, and 3mL samples were taken every 12 hours until the fourth day. The residual amount of (R,S)-DCPP was detected, the degradation rate was calculated, and the time-degradation curve of the chiral herbicide (R,S)-DCPP strain DCP-6 was drawn. Such as image 3 , strain DCP-6 can completely degrade 30mg / L (R,S)-DCPP within 96h.

[0047] HPLC detection of chiral herbicide (R,S)-DCPP...

Embodiment 3

[0056] Embodiment 3 bacterial agent preparation

[0057] The original seed of the chiral herbicide (R,S)-DCPP degrading strain DCP-6 of the present invention is activated on the test tube slope, and the degradation performance is measured, and then inoculated on the test tube slope for future use. The test tube seed was inoculated in a 1000mL shake flask containing 200mL LB medium (LB medium formula: peptone 10g, yeast powder 5g, sodium chloride 5g, water 1L, pH 7.4), cultured at constant temperature and shaken to the logarithmic phase, ready to inoculate one grade seed jars. The primary seed tank is 50L, the feeding volume is 40L, the medium formula is: glucose 8g / L, yeast extract 5g / L, K 2 HPO 4 1g / L, NaCl 5g / L, CaCO 3 2g / L, MgSO 4 0.2g / L, soybean oil 0.1% (v / v), pH 7.2-7.5.

[0058] After feeding, sterilize under high pressure at 121°C, cool to 30°C, inoculate 10% of the cultured shake flask strain into a 50L first-level seed tank, and cultivate until the logarithmi...

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Abstract

The invention discloses a degrading bacterial strain capable of simultaneously degrading two isomers of the chiral herbicide 2,4-dipropionic acid [(R,S)-DCPP] and the bacterial agent produced therefrom. Strain DCP‑6 was identified as Sphingopyxis sp., and was deposited in the China Center for Type Culture Collection on December 23, 2019, with the strain preservation number CCTCC M 20191084. The degrading bacterial strain DCP-6 described in the present invention can broadly degrade phenoxycarboxylic acid chiral herbicides (R, S)-DCPP and 2-methyl-4-chlorophenoxypropionic acid (R, S)-MCPP Two isomers and phenoxycarboxylic acid achiral herbicides 2,4-dichlorophenoxyacetic acid (2,4-D), MCPA and 2,4-DB, etc., degrading bacterial agent products can make soil (R,S)-DCPP and other residues are reduced by more than 89%, which can effectively solve the problems of phenoxycarboxylic acid herbicides in soil and water environment pollution and crop phytotoxicity in the agricultural production process, and protect the ecological environment.

Description

technical field [0001] The invention relates to a degrading bacterial strain capable of simultaneously degrading two isomers of the chiral herbicide 2,4-dipropionic acid and a bacterial agent produced therefor, belonging to the field of biological high technology. Background technique [0002] Chemical pesticides are an important guarantee for the rapid development of agriculture and food security in the world, and have played an inestimable role in the modernization of agricultural production. Pesticides with chiral structures (chiral pesticides) account for about 40% of the chemical pesticides currently in use in my country. At the same time, with the continuous development of chemical pesticide varieties, the number of pesticide chiral compounds has increased significantly. Among the major agricultural compounds, the usage of herbicides has ranked first. Studies have shown that herbicides with chiral structures (chiral herbicides) have enantiomeric differences that cann...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A62D3/02C12R1/01A62D101/04A62D101/28A62D101/22
CPCC12N1/20A62D3/02A62D2101/04A62D2101/28A62D2101/22C12R2001/01C12N1/205
Inventor 蒋建东张龙陈凯胡顺利
Owner NANJING AGRICULTURAL UNIVERSITY
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