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A natural ammonium-resistant nitrogen-fixing microorganism lq3 and its application

An ammonium-resistant and nitrogen-fixing technology, applied in microorganisms, microorganisms, microorganism-based methods, etc., can solve the problem of low nitrogen-fixing activity, and achieve the effects of high nitrogen-fixing activity, high-efficiency biological nitrogen-fixing, and broad application prospects.

Active Publication Date: 2021-02-05
CHINA AGRI UNIV
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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 nitrogen-fixing microorganism lq3 and its application
  • A natural ammonium-resistant nitrogen-fixing microorganism lq3 and its application
  • A natural ammonium-resistant nitrogen-fixing microorganism lq3 and its application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] The isolation, cultivation and identification of embodiment 1 Paenibacillus LQ3

[0035] 1. Isolation of Paenibacillus LQ3:

[0036] Paenibacillus LQ3 was isolated from rhizosphere soil samples of grapevines in Shuangtang Village, Shencun Town, Xuanzhou District, Xuancheng City, Anhui Province.

[0037] 2. Cultivation of Paenibacillus LQ3

[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; H 2 O was adjusted to 1L. Sterilize at 120°C for 20 minutes. The production of solid medium is to add 1.5g of agar per 100ml of liquid medium.

[0039] Paenibacillus LQ3 was inoculated into solid or liquid medium, and cultivated in a 30°C incubator for 2 days. The colony morphology of Paenibacillus LQ3 is as follows: figure 1 shown.

[0040] 3. Identification of Paenibacillus LQ3

[0041] (1) Extraction of total bacterial DNA: Genomic DNA wa...

Embodiment 2

[0054] The determination of the nitrogenase activity of embodiment 2 Paenibacillus LQ3 under different conditions

[0055] 1. Determination of Nitrogenase Activity

[0056] Inoculate Paenibacillus LQ3 in 5mL LD medium, culture overnight at 30°C, transfer to a 500mL Erlenmeyer flask at 1% inoculum size, culture at 30°C for 8 hours, then collect the bacteria, and use appropriate amount of medium with different ammonium concentrations (Add different concentrations of NH in 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 .

[0057] The base medium (1L) for measuring enzyme activity is: 26.3g Na 2 HPO 4 12H 2 O; 3.4 g KH 2 PO 4 ; 10 ...

Embodiment 3

[0073] Example 3 Growth-promoting effect experiment of Paenibacillus nitrogen-fixing bacteria LQ3 strain on cucumber

[0074] Centrifuge the bacterium liquid obtained from culturing in the basic medium for measuring enzyme activity to collect the bacterium, and then suspend it with deionized water to adjust its cell density OD 600 When it is 1.0, the bacterial suspension is obtained. The germinated plant seeds were soaked in the bacterial suspension for 30min, and then transplanted into small pots (sterile nutrient soil: vermiculite at a ratio of 1:1). After the plants grew for 2 weeks, each plant was irrigated with 15 mL of bacterial suspension. After 4 weeks of plant growth, the plants were harvested, the soil at the roots of the plants was washed with running water, and the length and dry weight of the plant roots and stems were measured. The deionized water treatment group was used as the control group, and the results are shown in Table 1.

[0075] Table 1 Growth-promot...

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Abstract

The invention relates to the technical field of nitrogen-fixing microorganisms, in particular to a natural ammonium-resistant nitrogen-fixing microorganism LQ3 and its application. The present invention provides a strain of Paenibacillus sp. LQ3, which is preserved in the General Microorganism Center of China Committee for Culture Collection of Microorganisms, and the preservation number is CGMCC No. 18073. The bacterium contains the nitrogenase structural gene nifH, which can maintain high nitrogen fixation activity and carry out efficient biological nitrogen fixation under high concentration ammonium (50-300mM) conditions, breaking the inhibition of high ammonium conditions on biological nitrogen fixation, and can be used in agricultural production Promote the use of fertilizers and effectively reduce the use of chemical fertilizers.

Description

technical field [0001] The invention relates to the technical field of nitrogen-fixing microorganisms, in particular to a natural ammonium-resistant nitrogen-fixing microorganism LQ3 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 action of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C05F11/08A01N63/25A01P21/00C12R1/01
CPCC05F11/08C12N1/205C12R2001/01Y02A40/10
Inventor 陈三风李琴
Owner CHINA AGRI UNIV
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