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Cadmium-resistant strain, method for constructing symbiont with microalgae and application of symbiont

A technology of symbionts and strains, which is applied in the field of cadmium-resistant strains and the construction of symbionts with microalgae, can solve problems such as limited tolerance of heavy metals, and achieve the effect of improving the removal effect.

Pending Publication Date: 2022-07-22
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these growth-promoting bacteria and symbionts have limited tolerance to heavy metals. To further improve the tolerance of bacteria to heavy metals and the effect of algae-bacteria symbionts on heavy metal water treatment, it is urgent to obtain microalgae that can symbiosis and have strong heavy metal resistant bacteria

Method used

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  • Cadmium-resistant strain, method for constructing symbiont with microalgae and application of symbiont
  • Cadmium-resistant strain, method for constructing symbiont with microalgae and application of symbiont
  • Cadmium-resistant strain, method for constructing symbiont with microalgae and application of symbiont

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Screening, purification and identification of cadmium-resistant bacteria

[0028] 1. Screening: Obtain fresh soil from the farmland near the lead-zinc-silver mining community in Qixia District, Nanjing, take 10g of fresh farmland soil sample and add it to a conical flask of 100mL sterile water, at 30℃, 180r·min -1 Condition, oscillate continuously for 12h and then stand for 1h;

[0029] 2. Purification: Take 100 μL of bacterial supernatant and spread it on a Cd-containing LB plate, place it in a constant temperature incubator for cultivation, and purify the grown single colony by streaking;

[0030] 3. Morphology and physiological characteristics analysis of the obtained strains:

[0031] (1) The morphology was observed by scanning electron microscope, and the specific steps were as follows: inoculate the obtained cadmium-resistant strains in LB broth medium, 30°C for 180r·min -1 After culturing for 24 hours, the bacterial cells were collected at 6000 rpm, washed 2-3 ...

Embodiment 2

[0048] Determination of Cd tolerance and removal ability of strain 2-23:

[0049] (1) The cadmium-resistant strains were inoculated into LB medium, and cultured in a shaker at 30° C. and 180 r / min for 24 h for activation. The activated bacteria were inoculated with Cd concentrations of 0, 5, 10, 20, 50 and 100 mg·L -1 100mL of LB medium was placed in a shaker for cultivation, and its initial OD 600 The values ​​are all around 0.1.

[0050] (2) Use a microplate reader to measure OD at 0, 1, 2, 6, 10, 14, 24, 36, and 48h of culture 600 value to construct a bacterial growth curve.

[0051] (3) The half-effect concentration EC50 value was used to express the toxicity of Cd to bacteria.

[0052] (4) Using the sigmoid dose-response curve equation to fit the OD of the strain grown at different concentrations for 24h 600 value, and then calculate the EC50 value of Cd on bacteria.

[0053] (5) The bacterial supernatant was collected at 0, 1, 2, 6, 10, 14, 24, 36, and 48 h, and th...

Embodiment 3

[0059] Construction of Phaeodactylum tricornutum-strain 2-23 algal-bacterial symbionts and Cd removal:

[0060] 1. Bacterial and Microalgae Suspension

[0061] (1) Cultivate Phaeodactylum tricornutum to logarithmic growth phase, centrifuge and discard the supernatant under centrifugal force of 8000g, wash 2-3 times with f / 2+Si, and collect the sediment in fresh sterile f / 2+ In Si, the microalgae suspension was prepared after mixing.

[0062] (2) Cultivate strains 2-23 to logarithmic growth phase, centrifuge and discard the supernatant under the condition of centrifugal force of 6000 rpm, wash 2-3 times with f / 2+Si, and collect the sediment in fresh sterile f / In 2+Si, the bacterial suspension was prepared after mixing.

[0063] 2. Proportion of microalgae and bacterial cells

[0064] (1) The wavelength of the OD value of the microalgae suspension was measured at 450 nm, and the number of cells in 1 mL of algal solution was calculated using the relationship between OD and th...

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Abstract

The invention discloses a cadmium-tolerant strain, a method for constructing a symbiont with microalgae and application of the cadmium-tolerant strain. The strain is screened from soil in a mining area, has relatively strong cadmium tolerance, and belongs to aeromonas veronii through 16SrDNA comparison. The microbe can be constructed with phaeodactylum tricornutum to form a phycomycete symbiont, and according to different bacterial proportions in the phycomycete symbiont, it is found that growth of the phycomycete symbiont with the high bacterial proportion and removal of heavy metal cadmium (Cd) are far superior to those of single microalgae.

Description

technical field [0001] The invention belongs to the technical field of heavy metal water treatment, and in particular relates to a cadmium-resistant strain, a method for constructing a symbiosis with microalgae, and its application. Background technique [0002] With the development of industrial production, human activities such as mining, metal smelting, and fossil burning have brought many heavy metals into the environment. Cadmium (Cd) is a highly toxic and carcinogenic heavy metal with very low limits allowed in the environment. In the environment, cadmium can enter human or animal body from air, water and food, and cause harm to the kidney, liver, bone and reproductive organs of human and animal body through amplification and bioaccumulation. In recent years, cadmium and its compounds have caused river water pollution in large areas, which has seriously threatened water quality and safety and human health. [0003] At present, the research focus on the removal of hea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/12C02F3/32C12R1/01C12R1/89C02F101/20
CPCC12N1/20C12N1/12C02F3/325C02F2101/20
Inventor 葛滢于清男裴欣雨韦炎平陈家乐相满城汪思婷常梦瑄
Owner NANJING AGRICULTURAL UNIVERSITY
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