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A Natural Ammonium-resistant Paenibacillus Nitrogenfixing Strain ah-4 and Its Application

A Bacillus, AH-4 technology, applied in bacteria, microorganisms, microorganisms and other directions, can solve the problem of low nitrogen fixation activity, and achieve the effect of high nitrogen fixation activity, efficient biological nitrogen fixation, and wide application prospects.

Active Publication Date: 2022-02-01
HUBEI FORBON TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The nitrogen-fixing activity of the nitrogen-fixing microorganisms reported so far is low under the condition of high concentration of ammonium

Method used

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  • A Natural Ammonium-resistant Paenibacillus Nitrogenfixing Strain ah-4 and Its Application
  • A Natural Ammonium-resistant Paenibacillus Nitrogenfixing Strain ah-4 and Its Application
  • A Natural Ammonium-resistant Paenibacillus Nitrogenfixing Strain ah-4 and Its Application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Isolation, culture and identification of Paenibacillus AH-4

[0035] 1. Isolation of Paenibacillus AH-4

[0036] Paenibacillus AH-4 was isolated from arbor rhizosphere soil samples in Hefei City, Anhui Province.

[0037] 2. Culture of Paenibacillus AH-4

[0038] Preparation of medium: nitrogen-free liquid medium (1L): 20g sucrose; 12.06g K 2 HPO 4 ;3.4g KH 2 PO 4 ;0.2gMgSO 4 ·7H 2 O; 0.01g NaCl; 0.01g FeCl 3 ; 0.002g NaMoO 4 ·2H 2 O; add H 2 O Make up to 1L. Sterilize at 115°C for 30min. Solid medium was prepared by adding 1.5 g of agar per 100 ml of liquid medium.

[0039]Paenibacillus AH-4 was inoculated into solid or liquid medium and cultured in a 30°C incubator for 2 days. The colony morphology of Paenibacillus AH-4 on nitrogen-free medium is as follows figure 1 shown.

[0040] 3. Identification of Paenibacillus AH-4

[0041] (1) Extraction of total bacterial DNA: genomic DNA was extracted from the pure culture of strain Paenibacillus AH-...

Embodiment 2

[0053] Example 2 Determination of nitrogenase activity of Paenibacillus AH-4 under different ammonium concentration conditions

[0054] 1. Determination of nitrogenase activity

[0055] Paenibacillus AH-4 was inoculated into 5 mL of LD medium, cultured at 30 °C overnight, transferred to a 500 mL conical flask at 1% of the inoculum, and cultured at 30 °C for 8 h, then collected the bacteria, and added appropriate amounts of different ammonium concentrations. Different concentrations of NH were added to the basic medium for measuring enzyme activity 4 Cl) suspended bacteria, adjusted OD 600 to 0.4. Inoculate 4 mL of bacterial liquid into the anaerobic culture tube, use an air extraction device to extract air and fill with argon, inject 10% acetylene into the anaerobic tube, cultivate at 30 °C, and inject 100 μL of gas into a gas chromatograph every 2 hours to determine the ethylene content. .

[0056] The basic medium (1L) for measuring enzyme activity is: 26.3g Na 2 HPO 4...

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Abstract

The invention relates to the technical field of nitrogen-fixing microorganisms, in particular to a natural ammonium-resistant Paenibacillus nitrogen-fixing strain AH‑4 and its application. The present invention provides a strain of Paenibacillus sp. AH‑4, which is preserved in the General Microorganism Center of China Microbiological Culture Collection Management Committee, and the preservation number is CGMCC No.18766. The bacterium contains nitrogenase structural gene nifH, which can maintain a high nitrogen fixation activity under high concentration ammonium (50-300mM) conditions, breaking the inhibition of high ammonium conditions on biological nitrogen fixation, and can be popularized and used in agricultural production, effectively reducing The amount of fertilizer used.

Description

technical field [0001] The invention relates to the technical field of nitrogen-fixing microorganisms, in particular to a natural ammonium-resistant nitrogen-fixing Paenibacillus AH-4 and its application. Background technique [0002] Nitrogen is a macronutrient essential for plant growth. Nitrogen is abundant on the earth's surface, but most of it exists in the atmosphere in the form of inert nitrogen, which cannot be used by living organisms. In order to ensure the high yield of food crops, vegetables and fruits, a large number of chemical nitrogen fertilizers are needed. However, the long-term excessive application of chemical nitrogen fertilizer will lead to soil deterioration, environmental pollution and product quality degradation, hindering the sustainable development of agricultural economy. [0003] Biological nitrogen fixation refers to the process in which a few prokaryotic microorganisms reduce nitrogen in the air to ammonium under the action of nitrogenase in ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C05F11/08C12R1/01
CPCC12N1/20C05F11/08C12R2001/01C12N1/205
Inventor 陈三凤李琴
Owner HUBEI FORBON TECH
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