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A strain of nitrate-reducing bacteria, its culture method and application

A culturing method and nitrate technology are applied in the field of culturing and nitrate-reducing bacteria, can solve problems such as short time-consuming time, and achieve the effects of short time-consuming time, convenient operation and broad application prospect.

Active Publication Date: 2020-09-04
北京本农科技发展有限公司 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The present invention solves the aforementioned needs by providing a new nitrate-reducing bacterial strain and its application, the cultivation method of the nitrate-reducing bacterial strain, and can be used for salt tolerance and salt resistance, especially for biological nitrogen fixation and high-salt wastewater treatment. Governance, the method is green and environmentally friendly, low in cost, short in time, and easy to operate. Through this method of biological treatment, the strain has excellent halophilic characteristics, and the growth amount increases with the increase of the salt concentration. When the salt concentration is below 8%, there will be no problem of weakened strain viability or reduced tolerance, and the application prospect is very broad

Method used

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  • A strain of nitrate-reducing bacteria, its culture method and application
  • A strain of nitrate-reducing bacteria, its culture method and application
  • A strain of nitrate-reducing bacteria, its culture method and application

Examples

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Effect test

Embodiment 1

[0043] The separation and purification of nitrate-reducing bacteria and the culturing method specifically include the following steps:

[0044] 1) Prepare nutrient broth medium: peptone 8g / L, beef meal 4g / L, sodium chloride 3g / L, glucose 3g / L;

[0045] 2) Screening of strains: strains tolerant to high concentrations of NaCl were isolated from the water samples using the above-mentioned nutrient broth medium. The water samples for the isolation of bacterial species were obtained from water samples from the coastal saline-alkali land in Zhoushan, Zhejiang. The isolates can tolerate high concentrations of NaCl without acclimation;

[0046] 3) Cultivate and prepare strain seed solution in nutrient broth, 30°C, 150r / min, after culturing for 24h, inoculate with 5% inoculum to the same amount containing different NaCl concentrations (0%, 4%, 8%, 10%) , 15%) nutrient broth medium, shake culture at 30°C, 150r / min, take samples when the culture time is 2h, 5h, 10h, 15h, 20h, 25h, 36h,...

Embodiment 2

[0052] The culture method of nitrate-reducing bacteria specifically comprises the following steps:

[0053] 1) Prepare nutrient broth medium: peptone 12g / L, beef meal 1g / L, sodium chloride 6g / L, glucose 0.5g / L;

[0054] 2) Screening of bacterial species: the bacterial strain obtained by the separation and purification method of Example 1;

[0055] 3) Cultivate and prepare strain seed liquid in the nutrient broth of step 1), at 25°C, 150r / min, and cultivate for 48h;

[0056] 4) Inoculate the same amount of medium containing different NaCl concentrations (0%, 4%, 8%, 10%, 15%) with an inoculum volume of 7% by volume, 35°C, 150r / min shaking culture for 2d until stable At the same time, samples were taken at the same time, the OD600 absorbance of each culture solution was measured, and the growth of bacteria was measured by turbidimetric method. Taking the salt concentration as the abscissa and the absorbance as the ordinate, draw the salt tolerance curve of the strain. And mic...

Embodiment 3

[0059] The separation and purification of nitrate-reducing bacteria and the culturing method specifically include the following steps:

[0060] 1) Preparation of nutrient broth medium: peptone 10g / L, beef meal 3g / L, sodium chloride 5g / L, glucose 1g / L;

[0061] 2) Screening of bacterial species: the bacterial strain obtained by the separation and purification method of Example 1;

[0062] 3) Cultivate and prepare strain seed liquid in the nutrient broth of step 1), at 25°C, 150r / min, and cultivate for 48h;

[0063] 4) Inoculate the above-mentioned nutrient broth medium and the medium containing 8% NaCl concentration with an inoculum of 5%, respectively, take samples, carry out microscopic observation of the bacterial cells, and observe the bacterial cells when the bacterial cells are cultured at high salt concentration. morphological changes, see Figure 3-4 .

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Abstract

The invention relates to the field of microorganisms, in particular to nitrate reducing bacteria, a culture method and application. The bacteria strain is deposited at the General Microbiology Centerof the China Microbial Culture Collection Management Committeewith a preservation number of CGMCC No:16609; the preservation time is October 24, 2018. The bacteria strain has excellent halophilic properties, whena salt concentration is below 8%, the growth amount increases with the increase of the salt concentration, there is no problem that the activity of the bacteria strain is weakened or the tolerance is reduced, the bacteria can be applied to the high-salt sewagetreatment, wastewater biological nitrogen removal, purification of inorganic nitrogen in aquaculture water and the like, and canalso be applied to biological nitrogen fixation,nitrate in soil is reduced into ammonia nitrogen, the soil nitrogen loss is reduced, the bacteria has the effects of saving a fertilizer, improving crop quality and the like, and an application prospect is very broad.

Description

technical field [0001] The invention relates to the field of microorganisms, in particular to a nitrate-reducing bacteria, a culture method and applications. Background technique [0002] Nowadays, the carbon-nitrogen ratio of land in my country is seriously out of balance. For half a century, chemical fertilizers have been used, and organic fertilizers are rarely used. Due to the imbalance of carbon-nitrogen ratio, soil nitrogen-fixing capacity decreases, 70% of nitrogen fertilizer enters the environment, and finally enters groundwater, air, and solidifies in the soil, causing huge waste and harm. The organic matter content of the soil itself is too low, below 1% on average; because the soil lacks organic matter, the soil loses the ability to retain water and fertilizer, resulting in serious soil erosion. [0003] In addition, due to serious environmental pollution, a large amount of saline wastewater has been produced. At present, many conventional physical, chemical and ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C02F3/34C12R1/01
Inventor 韩建均梁雪杰赵川王玉文
Owner 北京本农科技发展有限公司
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