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Endophytic Burkholderia wj-19 from Sapium arvensis with high tolerance to multiple heavy metals

A Burkholderia, Holderia technology applied in the field of discovery and characterization of Burkholderia

Active Publication Date: 2021-05-11
HUNAN ACAD OF FORESTRY
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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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  • Endophytic Burkholderia wj-19 from Sapium arvensis with high tolerance to multiple heavy metals
  • Endophytic Burkholderia wj-19 from Sapium arvensis with high tolerance to multiple heavy metals
  • Endophytic Burkholderia wj-19 from Sapium arvensis with high tolerance to multiple heavy metals

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: Screening of heavy metal-tolerant Sapium arvensis endophytic bacteria

[0021] 1. Isolation and purification of endophytic bacteria from plants:

[0022] The fresh Sapium aconitifolia 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.

[0023] 2. The verification of the experimental effect of the surface disinfection of Chinese tallow tree, the specific implementation method is (one of the following two methods can be selected):

[0024] Method 1: Take 200 μL of washing water from the last step of surface disinfec...

Embodiment 2

[0026] Example 2: Biological identification and strain morphological characteristics of bacterial strains

[0027] 1. Morphological characteristics of the strain

[0028] (1) Morphological characteristics of colony population: Streak inoculation of WJ-19 monoclonal colony into LB solid medium, place the plate upside down in a constant temperature incubator, and incubate at 35°C for 18-24h, forming medium-sized, opaque, moist, raised Milky white colonies. Under the conditions of LB liquid culture (temperature 30°C, pH=6, shaker 170r / min), WJ-19 was in the lag phase at 0-4h, entered the logarithmic growth phase at 4-10h, and then entered the stable phase and decay phase.

[0029] (2) Individual morphological characteristics of the bacteria: Gram staining was carried out on the WJ-19 strain, and observation by an optical microscope showed that the strain was a rod-shaped, Gram-negative bacterium.

[0030] 2. Strain 16S rRNA analysis

[0031] Bacterial Genomic DNA Extraction Ki...

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, spot the same amount of fermentation broth of these four concentration gradients on the solid medium of LB strains added with heavy metals lead nitrate, zinc chloride, and cadmium chloride, and screen to obtain The colony WJ-19 that can grow on both.

[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 nitrate, zinc chloride, and cadmium chloride, an...

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Abstract

The invention belongs to the field of environmental management. The invention discloses a strain of Burkholderia sp. resistant to heavy metals and its effect on removing heavy metals. The strain is an endophytic bacterium Burkholderia sp isolated from tallow planted in heavy metal mining areas. .WJ‑19. The bacteria has a high tolerance level to lead, zinc, and cadmium, which are typical heavy metals in southern China, and has high removal efficiency for zinc and cadmium in aqueous solution, and can grow in culture solutions containing high concentrations of heavy metals zinc and cadmium without obvious inhibition. WJ‑19 provides a new material for microbial treatment of heavy metal pollution, which has potential for 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 Patents(China)
IPC IPC(8): C12N1/20C12N1/02C02F3/34C12R1/01C02F101/20C02F101/22
CPCC02F3/34C02F2101/20C02F2101/22C12N1/02C12N1/20C12N1/205C12R2001/01
Inventor 黄忠良朱咏华张轩刘选明李辉曾嘉锐吴子剑刘颖黄兢宁帅然覃晓莉刘则一柳倩
Owner HUNAN ACAD OF FORESTRY
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