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Screening of endosulfan degrading bacteria JW2 and application of endosulfan degrading bacteria JW2 to red soil

A technology for degrading bacteria and endosulfan, which is applied to the restoration of polluted soil, bacteria, and microorganism-based methods, etc., can solve the problem of high-efficiency endosulfan-degrading bacteria, etc., and achieves restoration of endosulfan-contaminated soil, good application prospects, and preparation of simple craftsmanship

Inactive Publication Date: 2014-05-14
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few reports on highly efficient endosulfan-degrading bacteria and its application in southern China.

Method used

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  • Screening of endosulfan degrading bacteria JW2 and application of endosulfan degrading bacteria JW2 to red soil
  • Screening of endosulfan degrading bacteria JW2 and application of endosulfan degrading bacteria JW2 to red soil
  • Screening of endosulfan degrading bacteria JW2 and application of endosulfan degrading bacteria JW2 to red soil

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1: Screening and Identification of Efficient Endosulfan Degrading Bacteria JW2

[0040] Medium:

[0041] Inorganic salt basal medium: 5.8g K 2 HPO 4 , 4.5g KH 2 PO 4 , 2.0g (NH 4 ) 2 SO 4 , 0.16g MgSO 4 , 0.02g CaCl 2 , 0.002g Na 2 MoO 4 , 0.001g FeSO 4, 0.001g MnCl 2 , 1L of deionized water, adjusted to pH 7.0, and sterilized at 121°C for 30min.

[0042] Medium containing a small amount of carbon source: Add 5.0 g of peptone to the above inorganic salt basic medium, adjust to pH 7.0, and sterilize at 121°C for 30 minutes.

[0043] Separation and purification medium: add endosulfan to the medium containing a small amount of carbon source, so that the concentration of endosulfan in the medium is 100 μg·mL -1 , 15.0 g of agar, adjusted to pH 7.0, and sterilized at 121 °C for 30 min.

[0044] LB medium: 10.0g peptone, 5.0g yeast extract, 10.0g NaCl dissolved in 1L deionized water, adjust the pH to 7.0, and sterilize at 121°C for 30min.

[0045] 1) S...

Embodiment 2

[0053] Example 2 Analysis of high-efficiency endosulfan-degrading bacteria JW216S rDNA sequence

[0054] 1. Extraction of total DNA of highly efficient endosulfan-degrading bacteria JW2

[0055] In the experiment, the MI BIO PowerSoilDNA Isolation Kit was used to extract the total DNA in the soil. The main operation steps were slightly improved on the basis of the original instructions. The specific process is as follows:

[0056] 1) Bacterial culture: Inoculate highly efficient endosulfan-degrading bacteria JW2 in LB medium, and culture at 30°C for 18 hours with shaking;

[0057] 2) Cell collection: Weigh and record the weight of the empty 10ml centrifuge tube. Take 5mL culture solution in a 10mL centrifuge tube, 8000r min -1 Centrifuge for 8 minutes, discard the supernatant, and collect the bacteria. Weigh and record the weight of the 10ml centrifuge tube containing the bacteria again, and the difference between the two weighing values ​​is the weight of the bacteria. Di...

Embodiment 3

[0086] Embodiment 3: Degradation characteristics of degrading bacteria JW2

[0087] Extraction of endosulfan from the separation and purification medium: Inoculate the high-efficiency endosulfan-degrading bacteria JW2 into the separation and purification medium and cultivate for 5 days, add 5mL of n-hexane, fully oscillate and extract on the vortex mixer, let it stand for stratification, and take the organic phase.

[0088]Determination of endosulfan: gas chromatographic analysis: Shimadzu GC-14C gas chromatograph, capillary column (OV-1701) 0.53mm×30m, FID detector. Injection port temperature 260°C, column temperature 240°C, detector 260°C, N 2 Flow rate 25mL·min -1 , the injection volume is 2 μL.

[0089] The formula for calculating the degradation rate of endosulfan by bacterial suspension:

[0090] R = ρ ck - ρ ρ ck ...

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Abstract

The invention provides screening of endosulfan degrading bacteria JW2 and application of the endosulfan degrading bacteria JW2 to red soil. The collection number of the endosulfan degrading bacteria JW2 is CCTCC No: M2012326. According to the characteristic that microorganisms can quickly degrade endosulfan and shorten the half-life period, the microorganisms capable of degrading the endosulfan are screened out under the slightly acidic and high-temperature environment and are applied to soil in the South of China, so that the aim of repairing the soil polluted by the endosulfan is fulfilled. The endosulfan degrading bacteria JW2 can degrade the residual endosulfan on water, soil and the like safely, efficiently and quickly so as to reduce the damage of the endosulfan to the environment and achieve the effects of repairing the soil polluted by the endosulfan and protecting the ecological environment. The endosulfan degrading bacteria JW2 has simple preparation process, low cost and high efficiency, avoids secondary pollution and has good application prospect.

Description

technical field [0001] The invention relates to the screening of an endosulfan-degrading bacterium JW2 and its application in red soil, belonging to the technical field of biodegradation treatment. Background technique [0002] Endosulfan (Endosulfan), the chemical name is 1,2,3,4,7,7-hexachlorobicyclo(2.2.1)heptene-(2)bishydroxymethyl-5,6-sulfite, the molecular formula is C9H6Cl6O3S, the molecular weight is 406.91. Technical endosulfan is composed of two specific isomers with a content greater than 95%. These two isomers are α-endosulfan and β-endosulfan, respectively, and the ratio of the two varies from 2:1 to 7:3. The structural formulas of α-endosulfan and β-endosulfan are as follows figure 1 Shown: [0003] [0004] Endosulfan is generally classified as a cyclopentadiene organochlorine insecticide, but it contains only one double bond. Technical grade endosulfan is a light brown to dark brown crystalline solid. The melting point range of α-endosulfan is 108-110...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20B09C1/10C12R1/01
Inventor 朱鲁生孔令芬王军王金花谢慧王凤花魏凯苏坤昌
Owner SHANDONG AGRICULTURAL UNIVERSITY
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