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A denitrifying bacterium with complete denitrification ability and rapid denitrification ability and its application

A technology of denitrification bacteria and denitrification, applied in the field of environmental microorganisms, can solve problems such as unclear denitrification rate

Active Publication Date: 2021-03-19
中国科学院遗传与发育生物学研究所农业资源研究中心
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Zhang et al. (2014) obtained the strain NEAU-ST5-5T from the saline-alkali soil of the Songnen Plain in China. The analysis pointed out that the strain is a new species of Pseudomonas, and it was named Pseudomonas songenen for the first time; through The conventional nitrate reduction test shows that the strain is positive, but it is not clear whether it can perform a complete denitrification process and how fast the nitrogen removal rate is.

Method used

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  • A denitrifying bacterium with complete denitrification ability and rapid denitrification ability and its application
  • A denitrifying bacterium with complete denitrification ability and rapid denitrification ability and its application
  • A denitrifying bacterium with complete denitrification ability and rapid denitrification ability and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Isolation and purification of denitrifying bacteria (Pseudomonas songnenensis) L103

[0024] Sample source: High nitrogen fertilizer application rate of 600kg(N) hm, Luancheng Agricultural Ecosystem Experimental Station, Agricultural Resources Research Center, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences -2 ·ya -1 Long-term positioning test farmland 100m aquifer soil.

[0025] TSA medium (solid): tryptone 15g / L, soybean peptone 5g / L, sodium chloride 5g / L, agar 15g / L, adjust the pH value to 7.2-7.4, sterilize by high pressure steam at 121°C for 30min.

[0026] TSB medium (liquid): tryptone 15g / L, soybean peptone 5g / L, sodium chloride 5g / L, adjust the pH value to 7.2-7.4, and sterilize under high pressure steam at 121°C for 30min.

[0027] LB medium (liquid): tryptone 10g / L, yeast extract 5g / L, sodium chloride 5g / L, adjust the pH value to 7.2-7.4, and sterilize by high pressure steam at 121°C for 30min.

[0028] Separation and...

Embodiment 2

[0029] Embodiment two: the identification of denitrifying bacteria (Pseudomonas songnenensis) L103

[0030] 1) Identification of biological characteristics

[0031] The colonies of denitrifying bacteria L103 were white protrusions with smooth edges; the cells were rod-shaped, about 1.5 μm in length and 0.5 μm in diameter, without flagella, but motile.

[0032]Motility detection (semi-solid puncture method): prepare semi-solid TSA medium (tryptone 15g / L, soybean peptone 5g / L, sodium chloride 5g / L, agar 7.5g / L) in a sterile 50mL centrifuge tube, use Dip a sterile toothpick to pick up an appropriate amount of the target bacteria, puncture and inoculate it into a semi-solid TSA medium, place it in a constant temperature incubator, and culture it overnight at 28°C, and use transmitted light for visual observation. As a result, it was observed that the denitrifying bacteria L103 diffused and grew in the form of clouds from the puncture line to the surroundings, but the denitrifying...

Embodiment 3

[0039] Example 3 Evaluation of denitrification ability of denitrifying bacteria (Pseudomonas songnenensis) L103

[0040] The denitrifying bacterium Pseudomonas songnenensis L103 of the present invention is facultative anaerobic bacterium,

[0041] The GC-Robot automatic sampling and determination system was used to evaluate its denitrification ability in anaerobic, microanaerobic, acidic and alkaline environments, and the results showed that the denitrifying bacteria L103 can completely reduce NO 3 - produce N 2 , with complete denitrification ability; in addition, it has denitrification ability in the range of pH 6.6-7.8, and the denitrification ability increases with the increase of pH value, and the maximum denitrification rate can be as high as 1.62-2.36g NO 3 - / day / L; Finally, the anaerobic and microanaerobic environments had little effect on the denitrification ability of denitrifying bacteria L103 in alkaline environment, and the denitrification process was relative...

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Abstract

The invention discloses a denitrifying bacterium Pseudomonas songnenensis L103 with complete denitrification ability and rapid denitrification ability and its application. The strain has been preserved in the China Center for Type Culture Collection on March 20, 2019, and the preservation number is CCTCC M 2019184. The strain has genes encoding nitrate reductase, nitrite reductase, nitric oxide reductase and nitrous oxide reductase. These genes can encode a complete denitrification enzyme system that catalyzes NO 3 ‑ Complete reduction yields N 2 , and the denitrification rate can be as high as 1.62~2.36g NO 3 ‑ / day / L. Therefore, the strain has important application value in the preparation of denitrifying microbial bacterial agent for reducing nitrate leaching in soil below the root layer and nitrate pollution in groundwater, as well as purification and treatment of nitrogenous sewage.

Description

technical field [0001] The invention belongs to the technical field of environmental microbes, and in particular relates to a denitrification bacterium with complete denitrification ability and fast denitrification ability and application thereof. Background technique [0002] In agricultural production, the amount of nitrogen fertilizer application far exceeds the demand of crops for nitrogen, resulting in the accumulation of soil nitrate. These accumulated nitrates can migrate downward with the water after encountering heavy rainfall, causing nitrate pollution in groundwater and seriously threatening human health. In addition, excessive nitrogen enters lakes and wetlands, causing a series of environmental problems such as eutrophication and deterioration of water quality. [0003] Denitrification is the microbial use of nitrate reductase (NAR), nitrite reductase (NIR), nitric oxide reductase (NOR) and nitrous oxide reductase (N 2 OR) and so on gradually change NO 3 - a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C02F3/34C12R1/38C02F101/16
CPCC02F3/34C02F2101/163C12N1/20C12N1/205C12R2001/38
Inventor 王新珍刘梦帅胡春胜刘彬彬
Owner 中国科学院遗传与发育生物学研究所农业资源研究中心
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