A Natural Ammonium-resistant Paenibacillus Nitrogenfixing Strain 23 and Its Application

A Bacillus, ammonium-resistant technology, applied in the natural ammonium-resistant Paenibacillus nitrogen-fixing 23 and its application field, can solve the problem of low nitrogen fixation activity, and achieve the effect of high nitrogen fixation activity, excellent nitrogen fixation activity, and high-efficiency biological nitrogen fixation

Active Publication Date: 2021-08-10
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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 23 and Its Application
  • A Natural Ammonium-resistant Paenibacillus Nitrogenfixing Strain 23 and Its Application
  • A Natural Ammonium-resistant Paenibacillus Nitrogenfixing Strain 23 and Its Application

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Experimental program
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Effect test

Embodiment 1

[0034] The isolation, cultivation and identification of embodiment 1 Paenibacillus 23

[0035] 1. Isolation of Paenibacillus 23

[0036] Paenibacillus 23 was isolated from alpine cypress rhizosphere soil samples in Beijing area.

[0037] 2. Cultivation of Paenibacillus 23

[0038] Production 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; plus H 2 O was adjusted to 1L. Sterilize at 115°C for 30 minutes. LD medium (1L): 5g yeast extract, 10g peptone, 2.5g NaCl. Sterilize at 121°C for 20 minutes. The production of solid medium is to add 1.5g of agar per 100ml of liquid medium.

[0039]Inoculate Paenibacillus 23 into solid or liquid medium and culture in a 30°C incubator for 2 days. The colony morphology of Paenibacillus 23 on nitrogen-free medium is as follows: figure 1 shown.

[0040] 3. Identification of Paenibacillus 23

[0041] (1) Extraction of...

Embodiment 2

[0053] The determination of nitrogenase activity of embodiment 2 Paenibacillus 23 under different ammonium concentration conditions

[0054] 1. Determination of Nitrogenase Activity

[0055] The Paenibacillus 23 obtained in Example 1 was inoculated in 5 mL of LD medium, cultivated overnight at 30°C, transferred to a 500mL Erlenmeyer flask at an inoculum size of 1%, and cultivated at 30°C for 8h, then collected the thalline, and used an appropriate amount of Medium with different ammonium concentrations (different concentrations of NH were added to the basic medium for measuring enzyme activity) 4 Cl) Suspended bacteria, adjust OD 600 to 0.4. Inoculate 4mL of bacterial liquid into the anaerobic culture tube, pump out the air with an aspirating device and fill it with argon, inject 10% acetylene into the anaerobic tube, cultivate at 30°C, take 100μL of gas every 2h and inject it into the gas chromatograph to measure the ethylene content .

[0056] The base medium (1L) for me...

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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 23 and its application. The present invention provides a strain of Paenibacillus sp. 23, which is preserved in the General Microorganism Center of China Committee for Culture Collection of Microorganisms, and the preservation number is CGMCC No. 18767. The bacterium contains the nitrogenase structural gene nifH, which can perform high-efficiency biological nitrogen fixation under high-concentration ammonium (50‑300mM) conditions, breaking through the inhibition of high ammonium conditions on biological nitrogen fixation, and can be popularized and used in agricultural production, effectively reducing chemical fertilizer usage.

Description

technical field [0001] The invention relates to the technical field of nitrogen-fixing microorganisms, in particular to a natural ammonium-resistant Paenibacillus nitrogen-fixing strain 23 and its application. Background technique [0002] Nitrogen is an essential macroelement 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 utilized by organisms. In order to ensure the high yield of food crops and vegetables and fruits, it is necessary to use a large amount of chemical nitrogen fertilizer. However, long-term excessive application of chemical nitrogen fertilizers will lead to soil deterioration, environmental pollution and product quality decline, hindering the sustainable development of agricultural economy. [0003] Biological nitrogen fixation refers to the process in which a small number of prokaryotic microorganisms reduce nitrogen in the air to ammonium under the acti...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/25A01P21/00C05F11/08C12R1/01
CPCA01N63/00C05F11/08C12N1/20C12N1/205C12R2001/01
Inventor 陈三凤李琴
Owner CHINA AGRI UNIV
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