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New application of Acidovorax LS-1, and kit for repairing arsenic pollution, and applications thereof

A new technology of acid-eating bacteria, applied in the field of environmental microorganisms, to achieve the effect of good application prospects

Active Publication Date: 2019-11-19
广东省科学院南繁种业研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Based on this, it is necessary to address the problem of arsenic pollution, provide a new application of acidovorous bacteria LS-1 and a kit for repairing arsenic pollution and its application

Method used

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  • New application of Acidovorax LS-1, and kit for repairing arsenic pollution, and applications thereof
  • New application of Acidovorax LS-1, and kit for repairing arsenic pollution, and applications thereof
  • New application of Acidovorax LS-1, and kit for repairing arsenic pollution, and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1, the separation and screening of acidovorous bacteria LS-1

[0038] Collect soil samples from the 10-20cm depth of the paddy field soil surface of the South China Institute of Botany, Chinese Academy of Sciences, Guangdong Province, and then place the collected soil samples and liquid selective medium in 100mL with a soil-water ratio of 1:100 (g / L). Vials were used for anaerobic culture. Introduce high-purity nitrogen for 30 minutes to maintain a fully anaerobic state in the system, then quickly press the rubber stopper and aluminum cover, and place it in a constant temperature anaerobic box at 30°C for cultivation in the dark. After culturing for one week, the liquid in the vial was transferred to a new liquid selection medium at a ratio of 1:10, and cultured under the above conditions, so that the inoculation and enrichment culture was repeated for 3-5 times.

[0039] In the anaerobic workstation, the enriched culture solution was diluted by a ten-fold g...

Embodiment 2

[0040] Example 2 strain identification

[0041] (1) Morphological identification

[0042]Judgment was made by observing the colony morphology of the screened strains on the solid plate medium, and the microscopic morphology of the colonies was observed with an optical microscope. Such as figure 1 As shown, the strain is short rod-shaped, about 1-2 μm long, about 0.3 μm wide, and has flagella.

[0043] (2) Molecular biological identification

[0044] Extraction of bacterial strain DNA: inoculate the preserved strains on the liquid selection medium, place in a constant temperature anaerobic box at 30°C for dark culture, take out the culture medium after 6 days of cultivation, centrifuge at 8000r / min for 5min, discard the supernatant to collect the precipitate bacteria. The total DNA of the isolated strains was extracted with the bacterial DNA extraction kit EZNATM Bacterial DNA Kit (Omega, USA), and the extracted DNA was stored at -20°C.

[0045] PCR amplification: the 16S ...

Embodiment 3

[0050] Embodiment 3, the tolerance characteristic of bacterial strain to AA (Ⅴ)

[0051] Acidovorax ebreus LS-1 was cultured in anaerobic LB medium to logarithmic phase, and then washed with sterilized liquid medium for 2-3 times to obtain a bacterial mother solution. Take an equal amount of bacterial mother solution and inoculate them into liquid media containing different AA(Ⅴ) concentrations (0, 200, 500, 1000 μM), take a sample every 24 hours to measure its OD600, record and draw the results under the conditions of different AA(Ⅴ) concentrations. OD600 curve over time. Among them, the liquid medium includes: Piperazine-N, N'-biA (2-ethaneAulfonic acid) (PIPES buffer pH 7.0 ± 0.2), 5mM sodium acetate, 1mL / L trace elements and 1mL / L vitamins.

[0052] Experimental results such as image 3 As shown, the results showed that the strain had strong arsenic tolerance, and the arsenic concentration was as high as 1mM.

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Abstract

The invention relates to a new application of Acidovorax LS-1, and a kit for repairing arsenic pollution, and applications thereof. The new application is the application of the Acidovorax LS-1 in repairing of arsenic pollution, wherein the Acidovorax LS-1 is added to an arsenic pollutant under anaerobic conditions to reduce pentavalent arsenic into trivalent arsenic, and the Acidovorax LS-1 has apreservation number of CGMCC NO: 11555. According to the present invention, the research results show that the Acidovorax LS-1 can reduce pentavalent arsenic into trivalent arsenic under anaerobic conditions, wherein the bioavailability and the mobility of the trivalent arsenic are much higher than the bioavailability and the mobility of the pentavalent arsenic, such that the activity of arsenicin the environment can be improved through the treatment so as to easily clear or reutilize the trivalent arsenic in the subsequent process; and with the application of the Acidovorax LS-1 in treatment of arsenic pollution, the pentavalent arsenic can be reduced into trivalent arsenic so as to be easily absorbed and enriched, such that the application has good application prospect in the repairingof arsenic pollution.

Description

technical field [0001] The invention relates to the technical field of environmental microorganisms, in particular to a new application of acidovorous bacteria LS-1, a kit for repairing arsenic pollution and the application thereof. Background technique [0002] Arsenic is a toxic substance that widely exists in soil, water and other natural environments. It not only reduces soil fertility, crop yield and quality, but also causes water pollution. The way people ingest arsenic is mainly through long-term drinking of drinking water contaminated with high concentration of arsenic and eating food containing arsenic. Through the action of the food chain, arsenic will eventually accumulate in the human body and seriously affect human health. [0003] The remediation technologies adopted for the current situation of arsenic pollution mainly include engineering remediation, physical and chemical remediation and microbial remediation. Due to the simple cultivation of microorganisms,...

Claims

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Application Information

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IPC IPC(8): C02F3/34C02F101/10
CPCC02F3/34C02F2101/103
Inventor 李爽陈迪文王庆吴启华敖俊华黄莹
Owner 广东省科学院南繁种业研究所
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