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Poplar endophyte burkholderia cenocepacia YG-3 with efficient tolerance to multiple heavy metals

A technology of Burkholderia and Holderia, applied in the field of discovery and characterization of Burkholderia, to achieve the effect of efficient removal of heavy metals

Active Publication Date: 2018-11-13
HUNAN ACAD OF FORESTRY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, it is of great purpose and selectivity to search for endophytic bacteria with strong tolerance to heavy metals from plants in heavy metal-polluted areas, but there are few reports on such endophytic bacteria so far.

Method used

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  • Poplar endophyte burkholderia cenocepacia YG-3 with efficient tolerance to multiple heavy metals
  • Poplar endophyte burkholderia cenocepacia YG-3 with efficient tolerance to multiple heavy metals
  • Poplar endophyte burkholderia cenocepacia YG-3 with efficient tolerance to multiple heavy metals

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: Screening of heavy metal-tolerant poplar endophytic bacteria

[0022] 1. Isolation and purification of endophytic bacteria from plants:

[0023] The fresh poplar samples were sterilized on the plant surface, dried and placed on mannitol soybean medium (soybean powder 20g / L, mannitol 20g / L, agar powder 20g / L, pH value 7.2±0.2), and cultured at 30°C , when the endophytic bacteria grow out, combined with streak separation method in LB solid medium (peptone 10g / L, yeast extract 5g / L, sodium chloride 10g / L, agar powder 18g / L, pH5.6-5.8 ), the temperature is 30°C, the culture time is 1-2 days, and the separation is carried out until the pure culture is obtained.

[0024] 2. The verification of the experimental effect of poplar surface disinfection, the specific implementation method is (one of the following two methods can be selected):

[0025] Method 1: Take 200 μL of washing water from the last step of poplar surface disinfection and evenly spread it on trypto...

Embodiment 2

[0027] Example 2: Biological identification and strain morphological characteristics of bacterial strains

[0028] 1. Morphological characteristics of the strain

[0029] (1) Morphological characteristics of colony population: Streak inoculation of YG-3 monoclonal colony into LB solid medium, place the plate upside down in a constant temperature incubator, and culture at 30°C for 18-24h, forming an opaque, moist, raised pale yellow colony. Under the condition of LB liquid culture (temperature 30°C, pH=6, shaker 170r / min), YG-3 is in the lag phase at 0-4h, enters the logarithmic growth phase at 4-10h, and then enters the stable phase and decay phase.

[0030] (2) Individual morphological characteristics of the bacteria: Gram-stained strain YG-3, observed with an electron microscope, the cell shape is rod-shaped with a length of about 1.2-1.6 μm and a width of about 0.5-0.6 μm, a Gram-negative bacterium .

[0031] 2. Strain 16S rRNA analysis

[0032] Bacterial Genomic DNA Ex...

Embodiment 3

[0038] Example 3: detection of strain heavy metal tolerance

[0039] The tolerance level of strains to cadmium, lead and zinc heavy metals was detected by dilution quantitative spotting method and plate coating method respectively, and the minimum inhibitory concentration of the strain was obtained.

[0040] 1. Colony dilution method to screen heavy metal-tolerant bacteria: OD 600 ≈1 strain fermentation broth, dilute it to OD with saline 600 ≈0.1, 0.01, 0.001, the same amount of the fermented broth of the four concentration gradients was applied to the solid medium of the LB strain added with heavy metals lead nitrate, zinc chloride, and cadmium chloride, and the four strains were obtained by screening. The colony YG-3 can grow in all concentrations.

[0041] 2. Determination of the minimum inhibitory concentration by plate coating: the microbial plate coating method was implemented on the solid medium of LB strains added with different concentrations of heavy metal lead nit...

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Abstract

The invention belongs to the field of environmental governance and discloses a strain of burkholderia cenocepacia tolerant to heavy metals and an effect of the burkholderia cenocepacia in removing heavy metals, wherein the strain is endophyte Burkholderia sp. YG-3 separated from the poplar root planted in a heavy metal mining area, and the collection number of the strain is CGMCC No.15105. The burkholderia cenocepacia YG-3 is highly tolerant to southern typical heavy metals lead, zinc and cadmium, and has high efficiency in removing zinc and cadmium in an aqueous solution, and can grow in a culture solution with high concentration of heavy metals zinc and cadmium without experiencing inhibition. The YG-3 provides a new material for microbial control of heavy metal pollution, and has potential in development and application.

Description

technical field [0001] The invention belongs to the field of environmental management and relates to the discovery and characteristics of a Burkholderia bacterium, in particular to the discovery and characteristics of a Burkholderia bacterium resistant to various heavy metals. Background technique [0002] In recent years, heavy metal pollution incidents have occurred frequently in my country, posing a serious threat to economic and social development, ecological environment construction, food safety and national health. The results of the national soil pollution survey released in 2014 showed that the total points of the country’s soil exceeding the standard rate were 16.1%, mainly inorganic models, accounting for 82.8% of all the exceeding points, mainly cadmium, mercury, arsenic, copper, 8 kinds of inorganic pollutants including lead, chromium, zinc and nickel. Hunan is the hometown of non-ferrous metals. The land area polluted by heavy metals such as lead, cadmium, merc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02C02F3/34C12R1/01C02F101/20C02F101/22
CPCC02F3/34C02F2101/20C02F2101/22C12N1/02C12N1/20C12N1/205C12R2001/01
Inventor 张轩朱咏华黄忠良刘选明李辉王湘吴子剑牛舒淇黄兢卜志刚覃晓莉姚晴晴董义
Owner HUNAN ACAD OF FORESTRY
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