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A Burkholderia bacterium resistant to heavy metal copper and its application

A technology of Burkholderia and Kholderia, which is applied in the application, bacteria, restoration of contaminated soil, etc., to achieve the effect of promoting stem length and plant height, and has broad application prospects

Active Publication Date: 2022-07-26
HANSHAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Wherein patent CN101671636B provides a kind of Burkholderia D54 that has heavy metal lead, cadmium, copper, zinc resistance, although the highest tolerance concentration of described Burkholderia D54 to copper ion is 150mg / L, but there is no report on the removal rate of copper ions in the environment and the growth-promoting effect on plants in high-copper-polluted environments

Method used

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  • A Burkholderia bacterium resistant to heavy metal copper and its application
  • A Burkholderia bacterium resistant to heavy metal copper and its application
  • A Burkholderia bacterium resistant to heavy metal copper and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Isolation and preservation of Burkholderia HXC-1

[0045] 1. Isolation and culture of Burkholderia HXC-1

[0046]Take mimosa (Mimosa pudica Linn.) collected from the suburbs of Chaozhou City, Guangdong Province, remove the stems and leaves, wash the roots with clean water, select a large, plump, fresh nodule at the root, wash away impurities, rinse well, and use After soaking in 95% ethanol for 5 min, use 0.1% HgCl 2 The surface was sterilized for 5 min, and then rinsed 10 times with sterile water under aseptic conditions. On the ultra-clean workbench after UV lamp sterilization, shake up the sterile water flowing down from the last rinse of the nodules, take a small amount of YMA medium for uniform coating, and invert at 28°C for 3 to 5 days. Observe the growth of the strain on the medium. If no colonies appear, it proves that the nodules have been cleaned and disinfected. Otherwise, the reasons for the colonies should be analyzed, and the nodules should b...

Embodiment 2

[0071] Example 2 Physiological and biochemical properties of Burkholderia HXC-1

[0072] 1.H 2 O 2 Enzyme activity assay

[0073] The described Burkholderia HXC-1 strain was inoculated on YMA solid medium, cultured at 28°C for 3 days, and 1 mL of 3% H 2 O 2 Drop on the bacterial moss, and each treatment is repeated three times; if bubbles appear within 5min, it is positive, and no bubbles are negative. The result is as image 3 As shown, the bacterial moss was added with H 2 O 2 After that, more bubbles were produced quickly, indicating that Burkholderia HXC-1 strain H 2 O 2 Enzyme activity is high.

[0074] 2. Congo red experiment

[0075] The Congo red test is used to detect the presence of cellulose in cell surface components. Congo red can stain cellulose into a red complex, so the colony turns red, which is a positive reaction, indicating that there is cellulose on the cell surface; the colony color does not change, it is a negative reaction, and there is no or...

Embodiment 3

[0092] Example 3 Determination of tolerance of Burkholderia HXC-1 to metallic copper

[0093] (1) Activation of Burkholderia HXC-1: First, streak the Burkholderia HXC-1 strain on solid YMA medium for inoculation, and then inoculate it in an incubator at a temperature of 28°C Cultivate for 3 days until the moss is full;

[0094] (2) Preparation of bacterial suspension: in the cultured plump bacterial lawn, use an inoculation loop to select a ring of colonies in 80 mL of YMA liquid medium (250 mL conical flask), and use a shaker (rotation speed is 120 r / min, temperature is 28 ℃) fully shaken for 15h to obtain bacterial suspension;

[0095] (3) Determination of the highest tolerated concentration of copper

[0096] Configure YMA solid medium containing copper (0, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600 mg / L) for use;

[0097] Take 1 mL of the bacterial suspension prepared in step (2) and add it dropwise to the above-mentioned YMA solid medium containing coppe...

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Abstract

The invention belongs to the technical field of microorganisms, and in particular relates to a heavy metal-resistant Burkholderia and application thereof. The invention provides a Burkholderia sp. HXC-1, the highest tolerance concentration of the Burkholderia HXC-1 to copper is greater than 150mg / L, and can effectively remove The removal rate of copper in the heavy metal polluted environment is as high as 92.57%, which can realize the remediation of heavy metal polluted soil; the Burkholderia HXC-1 has a growth promoting effect on the growth of alfalfa seedlings under copper pollution, and can significantly promote the growth of alfalfa seedlings. Stem length and plant height of alfalfa seedlings, and when the copper concentration was 450 mg / L, after the alfalfa seedlings were treated with the described Burkholderia HXC-1, the stem length and plant height were untreated 46 times and 2.89 times; when the copper concentration was 500mg / L, the alfalfa seedlings were treated with the described Burkholderia HXC-1, and the stem length and plant height were 67.96 times, 2.44 times, has a significant growth-promoting effect, and has broad application prospects.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a heavy metal-resistant Burkholderia and application thereof. Background technique [0002] In recent years, the development of industry and human activities have led to increasingly serious heavy metal pollution in soil. Heavy metals cannot be decomposed by themselves in the soil, the residual time is long, the harm is great, and they are easily absorbed by crops and affect human health. Copper pollution is a common heavy metal pollution. The mining of copper mines, the accumulation of tailings, the discharge of industrial wastewater, the use of copper-containing pesticides and other factors will lead to excessive accumulation of copper in the soil. The copper element is absorbed by the plant and stays in the root, which hinders the absorption of other components, especially iron, by the root system, resulting in poor plant growth. And too much copper into t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/20A01P21/00B09C1/10C12R1/01
CPCA01N63/20B09C1/10
Inventor 王桔红陈文
Owner HANSHAN NORMAL UNIV
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