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A kind of atrazine-degrading bacteria and its application

A technology of atrazine and degrading bacteria, applied in the direction of bacteria, biological water/sewage treatment, microorganisms, etc., can solve the problems of food and drinking water safety, high cost and difficult implementation, and difficult treatment, etc., to achieve good degradation properties, good engineering application potential and application prospects, and the effect of high degradation efficiency

Inactive Publication Date: 2019-07-05
TONGJI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Contamination of Atrazine in Soil and Water Causes Food and Drinking Water Safety Issues
For the non-point source pollution of pesticides, due to its wide range of pollution and large area of ​​pollution, other methods of treatment are difficult and costly and difficult to implement

Method used

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  • A kind of atrazine-degrading bacteria and its application
  • A kind of atrazine-degrading bacteria and its application
  • A kind of atrazine-degrading bacteria and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Screening of CX-T strains: The strains capable of degrading atrazine were isolated from soil samples by means of enrichment culture and streak separation, and the degradation efficiency was preliminarily evaluated to screen out efficient degrading strains.

[0041]1. Reagents required: Atrazine standard (ATR): Dr.Ehrenstorfer, 99.8%; Atrazine original drug: TCI, 99.6%; Chromatographic pure reagents such as acetonitrile were purchased from Shanghai Anpu Company; yeast extract, peptone, Agar powder, etc. are domestic analytical reagents.

[0042] 2. Medium

[0043] Basal medium (MSM, g / L): K 2 HPO 4 ·3H 2 O 1.60g, KH 2 PO 4 0.40g, 3.00g sucrose, MgSO 4 ·7H 2 O 0.20g, NaCl 0.10g, CaCl 2 0.02g, atrazine (1g / L methanol solution) 10mL, trace elements 5mL, dH 2 O1000mL

[0044] LB medium: yeast extract 5g, peptone 10g, NaCl 5g, dH 2 O 1000mL

[0045] All the above mediums were steam sterilized at 121° C. and 0.1 MPa for 20 minutes, and 20 g of agar powder was add...

Embodiment 2

[0066] Identification of degrading strains

[0067] Main reagents: SPIN Kit for Soil(MP), Taq DNA polymerase, dNTPs, agarose, 18-T Vector Ligation Kit, Plasmid Extraction Kit, H 2 o 2 , crystal violet, safranin, oxidase, etc.

[0068] This example only conducts a simple analysis of the characteristics of the strains such as Gram staining, catalase, and oxidase. For specific principles, please refer to the "Common Bacterial System Identification Manual".

[0069] Genome extraction: Purified strains were inoculated with LB medium for overnight culture, according to SPIN Kitfor Soil Operation Manual Add the strain culture solution, and the template DNA can be obtained within 3-4 hours.

[0070] Physiological and biochemical characteristics of the bacterial strain were analyzed, as shown in Table 3, the physiological and biochemical identification results showed that CX-T and K were Gram-negative bacteria, catalase and oxidase-positive bacteria; H and W were Gram-positive,...

Embodiment 3

[0078] 1 / 10LB medium: yeast extract 0.5g, peptone 1.0g, NaCl 0.5g, dH 2 O 1000mL. The medium was sterilized by steam at 121°C and 0.1MPa for 20min. Other mediums are the same as above.

[0079] Analysis of degradation products: Firstly, the standard substances of cyanuric acid and hydroxyatrazine were optimized by ion source, the samples of the culture solution were directly passed through a 0.22 μm microporous membrane, and the optimized mass spectrometry conditions were used to directly inject samples to detect the target substances. group. The ion source is an ESI source.

[0080] 1. Determination of growth curve:

[0081] The bacterial suspension was inoculated into 1 / 10LB medium according to the inoculation amount of 1%, cultured at 30°C and 180r / min, and samples were taken every 4h for the first 28h to measure the OD 600 value, and each strain was replicated three times. At the same time, make a blank control.

[0082] 2. Determination of degradation curve:

[00...

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Abstract

The present invention relates to an atrazine-degrading bacterium and its application. The degrading bacterium belongs to the Rhizobium family, and is preserved in the China Center for Type Culture Collection. The preservation number is: CCTCC No: M2015741, and the preservation date is 2015 December 14th. The degrading bacterium has high-efficiency degradation characteristics for the pesticide atrazine, and is used for bioremediation of water bodies and soils polluted by the pesticide atrazine. In the bioremediation project, the bacteria can rapidly proliferate through cell division, and quickly establish a stable community. When growing in atrazine-contaminated soil and water, the pesticide atrazine can be used as the only nitrogen source to decompose atrazine. The biodegradation of atrazine achieves the purpose of controlling pesticide pollution. This strain can survive in soil and water, and is tolerant to high concentrations of atrazine. It has broad application prospects and good environmental benefits.

Description

technical field [0001] The invention relates to a bacterial strain, in particular to an atrazine-degrading bacterium and application thereof. Background technique [0002] It is estimated that 12.5 million tons of pesticide active ingredients are used in agricultural production every year in the world, among which herbicides are used in the largest amount, accounting for about 40% of the pesticide usage. Atrazine (2-chloro-4-2-isopropylamino-6-ethylamino-s-triazine) has the same active structure as the triazine herbicides simazine, terbutazine and permethazine, and is used in the largest amount Triazine herbicides are widely used in the agricultural production of corn, sorghum and other crops to control annual grasses and broadleaf weeds before and after emergence. Atrazine enters the environment primarily due to inappropriate use, weed control and subsequent surface runoff into surface water infiltration or through the soil into groundwater. The behavior of atrazine in th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20B09C1/10C02F3/34C12R1/01C02F101/38
CPCB09C1/10C02F3/34C02F2101/38C12N1/205C12R2001/01
Inventor 马利民陈松松杨盼盼
Owner TONGJI UNIV
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