Burkholderia sp. resistant to heavy metal copper and application

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

Active Publication Date: 2021-08-24
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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  • Burkholderia sp. resistant to heavy metal copper and application
  • Burkholderia sp. resistant to heavy metal copper and application
  • Burkholderia sp. resistant to heavy metal copper and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] The isolation and preservation of embodiment 1 Burkholderia HXC-1

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

[0046]Take Mimosa pudica Linn. collected from the suburbs of Chaozhou City, Guangdong Province, remove the stems and leaves, wash the roots with water, choose a large, plump, fresh root nodule from the roots, wash away impurities, rinse, and rinse with water. After soaking in 95% ethanol for 5 min, use 0.1% HgCl 2 The surface was sterilized for 5 minutes, and then rinsed 10 times with sterile water under sterile conditions. On the ultra-clean workbench after ultraviolet light sterilization, shake the sterile water flowing down from the root nodules for the last time, take a small amount to spread evenly in YMA medium, and incubate upside down at 28°C for 3 to 5 days. Observe the growth of the strain on the culture medium. If no bacterial colonies appear, it proves that the root nodules have been cleaned and disinfected; otherwise, the cause ...

Embodiment 2

[0071] Physiological and biochemical characteristics of embodiment 2 Burkholderia HXC-1

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

[0073] The 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 it on the bacterial lawn, and repeat each treatment three times; if bubbles appear within 5 minutes, it is positive, and if there are no bubbles, it is negative. The result is as image 3 As shown, the bacterial lawn added H 2 o 2 After that, more air bubbles were produced rapidly, indicating that Burkholderia HXC-1 strain H 2 o 2 Enzyme activity is high.

[0074] 2. Congo red experiment

[0075] The Congo red assay is used to detect the presence of cellulose in cell surface components. Congo red can dye cellulose into a red compound, so the colony turns red, which is a positive reaction, indicating that there is cellulose on the cell surface; if the color of the colony remains unchanged, it is a negative reac...

Embodiment 3

[0092] Example 3 Burkholderia HXC-1 Tolerance Determination of Metallic Copper

[0093] (1) Activation of Burkholderia HXC-1: first inoculate the Burkholderia HXC-1 strain on the solid YMA medium, and then inoculate it in an incubator with a temperature of 28°C. Cultivate for 3 days until the lawn is plump;

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

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

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

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

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Abstract

The invention belongs to the technical field of microorganisms, and particularly relates to Burkholderia sp. resistant to heavy metal and application. The invention provides Burkholderia sp. HXC-1, and the highest tolerance concentration of the Burkholderia sp. HXC-1 to copper is greater than 150 mg/L; the Burkholderia sp. HXC-1 can effectively remove copper in a heavy metal polluted environment, the removal rate reaches up to 92.57%, and remediation of heavy metal polluted soil can be achieved. The burkholderia HXC-1 has a growth promoting effect on growth of medicago sativa seedlings under copper pollution and can remarkably promote the stem length and plant height of the medicago sativa seedlings, and when the copper concentration is 450 mg/L, after the medicago sativa seedlings are treated with the burkholderia HXC-1, the stem length and the plant height of the medicago sativa seedlings are 46 times and 2.89 times of those of the medicago sativa seedlings which are not treated with the burkholderia HXC-1; when the copper concentration is 500mg/L and the burkholderia HXC-1 is used for treating medicago sativa seedlings, the stem length and the plant height of the burkholderia HXC-1 are 67.96 times and 2.44 times of those of the medicago sativa seedlings which are not treated by the burkholderia HXC-1, so the remarkable growth promoting effect is achieved and a good application prospect is achieved.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a Burkholderia resistant to heavy metals and its application. Background technique [0002] In recent years, the development of industry and human activities have resulted in increasingly serious heavy metal pollution in soil. Heavy metals cannot be decomposed by themselves in the soil, remain for a long time, cause great harm, and are easily absorbed by crops to affect human health. Copper pollution is a common heavy metal pollution. Factors such as copper mining, tailings accumulation, industrial wastewater discharge, and the use of copper-containing pesticides will cause excessive accumulation of copper in the soil. However, after the copper element is absorbed by the plant, it stays in the root, which hinders the root system from absorbing other components, especially the iron element, resulting in poor plant growth. Moreover, excessive copper entering th...

Claims

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Application Information

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Patent Type & Authority Applications(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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