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Endophytic Enterobacter yg-14 resistant to heavy metals

A technology of heavy metals and Enterobacteriaceae, applied in the direction of bacteria, microorganisms, microorganisms, etc.

Active Publication Date: 2021-07-06
HUNAN UNIV +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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  • Endophytic Enterobacter yg-14 resistant to heavy metals
  • Endophytic Enterobacter yg-14 resistant to heavy metals
  • Endophytic Enterobacter yg-14 resistant to heavy metals

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Screening of heavy metal-tolerant poplar endophytic bacteria

[0024] 1. Isolation and purification of endophytic bacteria from plants:

[0025] The fresh poplar samples were disinfected on the plant surface, dried and placed on a mannitol soybean medium (soybean powder 20g / L, mannitol 20g / L, agar powder 20g / L, pH value 7.2±0.2), cultured at 30°C , wait for the endogenous bacteria to grow, and then combine the 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 was 30 °C, the incubation time was 1-2 d, and the separation was carried out until the pure culture was obtained.

[0026] 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):

[0027] Method 1: Take 200 μL of the washing water from the last step of poplar surface disinfection and evenly s...

Embodiment 2

[0029] Example 2: Biological identification of strains and morphological characteristics of strains

[0030] 1. Morphological characteristics of strains

[0031] (1) Morphological characteristics of colony population: YG-14 monoclonal colonies were streaked into LB solid medium, the plate was placed upside down in a constant temperature incubator, and cultured at 30°C for 10-18 hours to form mucus-like yellow colonies. Under the condition of LB liquid culture (temperature 30°C, pH=6, shaker 170r / min), YG-14 was in the lag phase at 0-3h, entered the logarithmic growth phase at 3-18h, and then entered the stationary phase and the dying phase.

[0032] (2) Morphological characteristics of individual bacteria: Gram-staining of the YG-14 strain and observation with an optical microscope showed that the strain was short rod-shaped, Gram-negative bacteria.

[0033] 2. Analysis of strain 16S rRNA

[0034] Bacterial genomic DNA extraction kit (Shanghai Jierui Bioengineering Co., Ltd....

Embodiment 3

[0041] Example 3: detection of strain heavy metal tolerance

[0042] 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.

[0043] 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-14 that can grow in all concentrations.

[0044] 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 le...

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Abstract

The invention belongs to the field of environmental management. The invention discloses a strain of Enterobacter sp. YG‑14, which is tolerant to heavy metals and its effect on removing heavy metals. The bacteria is isolated from the roots of poplar trees planted in heavy metal mining areas. . The fungus 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 has a good removal effect on heavy metal lead of different concentrations when combined with poplar biochar. As a new microbial resource, YG‑14 has great development potential in biological treatment of heavy metal pollution.

Description

technical field [0001] The invention belongs to the field of environmental management, and relates to the discovery and characteristics of an Enterobacter, in particular to the discovery and characteristics of a heavy metal-tolerant Enterobacter. 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 national soil pollution rate was 16.1%. 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, mercury and arsenic reaches 28,000 square kilometers, accounting for 13% of the total area of ​​the province. The polluted mine land reaches 2.6 million mu, and the amount of polluted soi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N1/02C02F3/34C12R1/01C02F101/20
CPCC02F3/34C02F2101/20C12N1/02C12N1/20C12N1/205C12R2001/01
Inventor 朱咏华刘选明张轩黄忠良徐婷李辉高艳吴子剑黄兢覃晓莉黄梅曹力丹喻艳南
Owner HUNAN UNIV
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