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Heavy metal tolerant populus tremula endogenous enterobacter YG-14

A technology of heavy metals and Enterobacteriaceae, applied in the direction of bacteria, microorganisms, microorganisms, etc., to achieve the effect of efficient removal of heavy metals

Active Publication Date: 2018-11-06
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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  • Heavy metal tolerant populus tremula endogenous enterobacter YG-14
  • Heavy metal tolerant populus tremula endogenous enterobacter YG-14
  • Heavy metal tolerant populus tremula endogenous enterobacter YG-14

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 sterilized on the plant surface, dried and placed on the mannitol soybean medium (soybean powder 20g / L, mannitol 20g / L, agar powder 20g / L, pH value 7.2±0.2), and cultivated 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.

[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 washing water from the last step of poplar surface disinfection and evenly spread it on ...

Embodiment 2

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

[0030] 1. Morphological characteristics of the strain

[0031] (1) Morphological characteristics of colony population: Streak inoculation of YG-14 monoclonal colony into LB solid medium, place the plate upside down in a constant temperature incubator, and incubate 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 is in the lag phase at 0-3h, enters the logarithmic growth phase at 3-18h, and then enters the stable phase and decay phase.

[0032] (2) Individual morphological characteristics of the bacteria: Gram staining was carried out on the YG-14 strain, and observation by an optical microscope showed that the strain was a short rod-shaped, Gram-negative bacterium.

[0033] 2. Strain 16S rRNA analysis

[0034] Bacterial Genomic DNA Extraction Kit (Shanghai Jierui Bioeng...

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 environment treatment and discloses a heavy metal tolerant enterobacter and a heavy metal removal effect thereof. The bacterium is a plant endogenous Enterobactersp. YG-14 which is separated from a populus tremula root planted in a heavy metal mine. The bacterium has a high tolerance level for type heavy metals such as lead, zinc and cadmium in the south areaof China, has high removal efficiency upon zinc and cadmium in water solutions, and has a good removal effect on the heavy metal lead of different concentrations after being compounded with populus tremula charcoal. As a novel microorganism resource, YG-14 has great development potential in biological treatment on 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 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, mercury and arsenic amounts to 28,00...

Claims

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Application Information

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Patent Type & Authority Applications(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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