Enrichment medium for nitrate dependent ferrous oxidizing flora, preparation method and application

An enrichment culture and nitrate technology is applied in the field of nitrate-dependent ferrous oxidizing bacteria enrichment medium and the preparation field of nitrate-dependent ferrous oxidizing bacteria enrichment medium, which can solve the problem of small specific gravity, inability to be comprehensive, Accurately reflect the distribution types and environmental effects of nitrate-dependent iron oxidizing bacteria, achieving high accuracy, avoiding oxygen interference with ferrous oxidation, and maintaining good species diversity

Inactive Publication Date: 2018-01-09
WUHAN TEXTILE UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although nitrate-dependent iron-oxidizing bacteria have been isolated and identified successively in soil, sediment, and water bodies, the proportion of microorganisms isolated by traditional culture in the total microbial community is very small, which cannot fully and accurately reflect the real Distribution types and environmental effects of nitrate-dependent iron-oxidizing bacteria in soil under different conditions

Method used

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  • Enrichment medium for nitrate dependent ferrous oxidizing flora, preparation method and application
  • Enrichment medium for nitrate dependent ferrous oxidizing flora, preparation method and application
  • Enrichment medium for nitrate dependent ferrous oxidizing flora, preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] A culture medium for enriching nitrate-dependent ferrous oxidizing bacteria in lake water, including conventional components and special components, wherein the conventional components are: calcium chloride, potassium chloride, magnesium chloride, ammonium chloride, potassium dihydrogen phosphate , SL-10, PIPES and vitamin B1 solution; special components are: ferrous sulfate, sodium nitrate and sodium acetate.

[0058] A kind of nitrate dependent ferrous oxidizing bacteria group enrichment medium, it is made up of the raw material of following weight portion (1L enrichment medium contains):

[0059] Calcium chloride: 0.5mM; potassium chloride: 0.3mM; magnesium chloride: 0.55mM; ammonium chloride: 1.2mM; potassium dihydrogen phosphate: 0.2mM; sodium acetate: 6.0mM; SL-10: 3.5ml; PIPES: 6.0 mM; Ferrous Sulfate: 13mM; Sodium Nitrate: 13.0mM; Vitamin B 1 : 6ml.

[0060] A method for preparing a culture medium for enriching and cultivating nitrate-dependent ferrous oxidizi...

Embodiment 2

[0071] A kind of enrichment medium for nitrate-dependent ferrous oxidizing bacteria group, it is made 1L culture solution by the raw material of following parts by weight:

[0072]

[0073]

[0074] Its preparation steps are identical with embodiment 1.

[0075] Through the above technical measures, a variety of nitrate-dependent iron-oxidizing bacteria were enriched in the two typical soils of Quaternary red soil and calcareous fluvo-aquic soil in Hubei Province, and there were 7 genera with the abundance of such bacteria greater than 1.0%. After two weeks of enrichment culture, the ferrous oxidation ability of this kind of microorganisms in the above-mentioned several culture solutions reached 72.44%-86.58%, which is consistent with the publicly reported literature Eric E. Roden, Dmitri Sobolev, Brian Glazer, et al. Potential for MicroscaleBacterial Fe Redox Cycling at the Aerobic-Anaerobic Interface[J].Geomicrobiology Journal, 2004,21(6):379-391) is relatively close, ...

Embodiment 3

[0077] A kind of enrichment culture application of nitrate dependent ferrous oxidizing bacteria group enrichment medium in rice soil, its steps are:

[0078] (1) Optimization of enrichment conditions:

[0079] Take 10g of soil sample and add it to 100ml of sterilized medium. The volume of the serum bottle is 125ml; 2 , H 2 and CO 2 Composition, its components are 80:10:10, v / v, and the culture time is 2 weeks; after the culture is over, take 10ml of the supernatant and add it to 90ml of the sterilized medium for subculture and enrichment culture, and subculture according to this method Enriched for three generations;

[0080] (2) Sample preparation:

[0081] Take 5ml of the third-generation enriched culture solution in step (1), add it to 95ml of the sterilized medium, and continue to cultivate for 336h, which is a sample, wherein a part of the sample is used to determine the reaction of iron-oxidizing bacteria to nitrate and B The matrix utilization ability of acid salt ...

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Abstract

The invention discloses an enrichment medium for nitrate dependent ferrous oxidizing flora, a preparation method and an application. The enrichment medium is prepared from calcium chloride, potassiumchloride, magnesium chloride, ammonium chloride, monopotassium phosphate, sodium acetate, SL-10, piperazine-1,4-bisethanesulfonic acid, ferrous sulfate, sodium nitrate and vitamin B1 in certain ratio.The preparation method comprises steps as follows: (1), preparation of the medium: a solution A, a solution B and a solution C are prepared; (2), sterilization of the medium: a sterilization method for the solution A and the solution C comprises the step that suction-filtration sterilization is performed by a polyether sulfone membrane under the aseptic condition, and a product is standby; (3), sterilization of the solution B: the sterilized solution A, solution B and solution C are sequentially poured into a culture flask for subpackaging, and a sterile culture solution is obtained. Microbial genetic information obtained after three generations or more of enrichment subculture and high-throughput sequencing is high in flux and specificity, defects of a traditional screening culture method are overcome, and information of nitrate dependent ferrous oxidizing flora in the environments such as soil, sediment, underground water and the like is disclosed comprehensively and accurately.

Description

Technical field: [0001] The present invention relates to the field of soil ecological environment, more specifically to a nitrate-dependent ferrous oxidizing bacteria group enrichment medium, and also relates to a preparation method for the nitrate-dependent ferrous oxidizing bacteria group enrichment medium in sediments and water bodies The invention also relates to the use of a nitrate-dependent ferrous oxidizing bacteria group enrichment medium. Background technique [0002] Iron is the fourth largest element in the earth's crust, and it is also an active variable price element in the soil. The presence of iron oxides has an important impact on the participation of microorganisms in soil, sediment and other environments in biogeochemical cycle processes. Under anaerobic conditions, soil iron oxidation can provide electron donors for some chemolithotrophic microorganisms; also under anaerobic conditions, short-chain fatty acids such as formate, acetate, and propionate can...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12Q1/24C12Q1/689
Inventor 彭其安胡荣桂伍玉鹏陈凌玉赵晖王布匀
Owner WUHAN TEXTILE UNIV
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