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Staphylococcus warneri and application thereof

A Staphylococcus, Warrenella technology, applied in Staphylococcus Wardenii and its application fields, can solve the problems affecting soil porosity, water holding capacity, reducing soil copper activity, a large amount of cost, etc., to achieve good restoration potential, increase pH value, The effect of reducing the content

Active Publication Date: 2017-04-26
SINOSTEEL MAANSHAN INST OF MINING RES +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Physical methods include soil replacement, leaching and other measures, which can reduce soil copper content in a short period of time, but require a lot of cost and are likely to cause secondary pollution
Chemical methods include adding amendments such as minerals and organic matter, which can reduce soil copper activity, but large-scale application may affect soil porosity, water holding capacity, pH and other physical and chemical properties

Method used

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  • Staphylococcus warneri and application thereof
  • Staphylococcus warneri and application thereof
  • Staphylococcus warneri and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] 1. Material preparation

[0031] (1) Bacterial source: isolated from the copper mine in Ma'anshan

[0032] (2) LB medium: tryptone 10.0g / L; yeast powder 5.0g / L; sodium chloride 10.0g / L; the medium was sterilized at 121°C for 20min; % agar.

[0033] (3) Copper-contaminated soil samples: Jiangxi Dexing Copper Mine dump soil samples, Dongguashan soil samples, dandelion planted soil, sorrel soil and copper grass soil, the copper content ranges from 125 to 18100 mg / kg.

[0034] 2. Screening of bacteria

[0035] 1) Screening of Staphylococcus varreria CCTCC NO:M 2016599 (copper-resistant bacteria)

[0036] 1) Extraction of soil bacteria: Take 1 g of the soil collected from the surrounding copper mine in Ma'anshan and add it to 9 mL of sterile water for suspension. The culture condition is 30°C, the shaking table speed is 180 r / min, and cultured for 24 hours.

[0037] 2) Preliminary screening of copper-resistant bacteria: configure bacterial basal medium (LB), add copper s...

Embodiment 2

[0040] Example 2: Identification of Bacteria

[0041] 1. Extraction of bacterial total DNA

[0042] Bacterial genomic DNA was extracted using a bacterial genomic DNA extraction kit (Nanjing Huapu Biotechnology Co., Ltd.), and the operation steps were followed in the instructions.

[0043] 2. PCR amplification of the target gene fragment

[0044] Using 16SrRNA sequence analysis method, PCR amplification uses bacterial 16SrRNA universal primers:

[0045] 27F (5'-AGAGTTTGATCCTGGCTCAG-3');

[0046] 1492R (5'-GGTTACCTTGTTACGACTT-3'); PCR reaction system total 50μL:

[0047] The preparation of the reaction system of table 1 PCR

[0048]

[0049] 3. Add the PCR reaction system according to Table 1 into the PCR tube. After centrifuging and mixing the reaction solution, place it in the PCR reactor. The PCR operation program is: ① Pre-denaturation at 94°C for 10 minutes; ② Denaturation at 94°C for 45 seconds; ③ Annealing at 55°C for 45 seconds. ; ④ Extend at 72°C for 90 s; ⑤ Rep...

Embodiment 3

[0058] Embodiment 3: the impact of Staphylococcus varreria CCTCC NO:M 2016599 of the present invention on soil pH and soil available copper

[0059] 1. Determination of some physical and chemical properties of the tested soil

[0060] 1) Soil pH measurement: 10g soil samples (Jiangxi Dexing Copper Mine dump soil samples, winter melon mountain soil samples, planted dandelion soil, sorrel soil and copper grass soil) add 25mL pure water, shake for 2h, let stand for half an hour , measured with a pH meter.

[0061] 2) Determination of soil organic matter

[0062] Reagent: 2A, potassium dichromate solution (K 2 Cr 2 o 7 )=0.8000mol / L: add 400mL water to 39.2245g, heat to dissolve, and dilute to 1L after cooling.

[0063] 2B, concentrated sulfuric acid (analytically pure, concentration 98%, density 1.84g / cm 3 )

[0064] 2C. Organic carbon standard solution: dilute 1.375g of glucose to 100mL.

[0065] Measuring method: Weigh air-dried soil samples that pass through a 100-mesh...

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Abstract

The invention discloses staphylococcus warneri classified and named as staphylococcus warneri KR809427. The staphylococcus warneri KR809427 is preserved in the China Center for Type Culture Collection on October 31, 2016, and a preservation number is CCTCC NO:M 2016599. The invention further discloses application of staphylococcus warneri to increase of soil pH value and passivation of soil copper activity and discloses an application bacterial agent. The staphylococcus warneri CCTCC NO:M 2016599 grows fast and is high in resistance to copper, acids, alkalis and salts and capable of remarkably increasing culture medium pH value and reducing effective copper content of soil, thereby being high in potential for remediation of different types of copper contaminated soil.

Description

technical field [0001] The invention belongs to the field of environmental application microorganisms, and in particular relates to a staphylococcus varreria and its application; in particular, it relates to the application in the restoration of copper-contaminated soil. Background technique [0002] Copper ore is an important metal mineral with a long history of development and provides a large amount of resources for industrial development. However, due to insufficient environmental protection measures in the past, the surrounding environment was polluted during copper mining and smelting. Copper has become the main pollutant in the soil around the mining area, destroying the environmental quality of the area and endangering ecology and human health. Therefore, the control of copper pollution is still one of the hot research issues in the world. [0003] So far, the methods of soil copper pollution control include physical methods, chemical methods, biological methods and...

Claims

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Application Information

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IPC IPC(8): C12N1/20B09C1/10C12R1/01
CPCB09C1/10C12N1/20C12N1/205C12R2001/01
Inventor 王飞杨帅吴将有葛滢常前发常勤明程一松朱桐王岐川马贵党李崇华
Owner SINOSTEEL MAANSHAN INST OF MINING RES
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