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Degradation strain capable of simultaneously degrading two isomers of chiral herbicide dichlorprop and bacteria produced from degradation strain

A technology of herbicides and isomers, which is applied in the field of degrading strains and the bacterial agents produced by them, can solve non-target biological toxicity and other problems

Active Publication Date: 2020-07-07
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • Degradation strain capable of simultaneously degrading two isomers of chiral herbicide dichlorprop and bacteria produced from degradation strain
  • Degradation strain capable of simultaneously degrading two isomers of chiral herbicide dichlorprop and bacteria produced from degradation strain
  • Degradation strain capable of simultaneously degrading two isomers of chiral herbicide dichlorprop and bacteria produced from degradation strain

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Experimental program
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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. like image 3 , strain DCP-6 can completely degrade 30mg / L (R,S)-DCPP within 96h.

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

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 degradation strain capable of simultaneously degrading two isomers [(R,S)-DCPP] of chiral herbicide dichlorprop and bacteria produced from the degradation strain. Strain DCP-6 is identified as Sphingopyxis sp., and is preserved at China Center for Type Culture Collection on December 23, 2019, and the strain preservation number is CCTCC M 20191084. The degradation strain DCP-6 described in the invention can degrade phenoxy carboxylic acid type chiral herbicides (R,S)-DCPP and 2-methyl-4-chlorophenoxypropionic acid (R,S)-MCPP and phenoxycarboxylic acid achiral herbicides 2,4-dichlorophenoxyacetic acid (2,4-D), MCPA and 2,4-DB in a broad spectrum. The degradation strain product can reduce the residual amount of the (R,S)-DCPP and the like by 89% or more, can effectively solve the problems that phenoxycarboxylic acid herbicides pollute and damage crops in soil and water body environment during agricultural production, 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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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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