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Salt-tolerant arthrobacter and nitrogen fixation application thereof

An Arthrobacter and salt-tolerant technology, applied in the direction of bacteria, organic fertilizers, microorganisms, etc., can solve problems such as increased use of chemical fertilizers, decreased soil fertility, and decreased enzyme activity, to achieve low operation and maintenance costs, reduce the amount of chemical fertilizers used, Nitrogen fixation capacity and long-lasting effect

Pending Publication Date: 2021-12-31
BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Due to the excessive application of chemical fertilizers, it will lead to soil pollution, accumulation of toxic substances, changes in soil biological communities, and reduction of animal species and numbers that are beneficial to the soil, resulting in soil compaction, damage to the aggregate structure, insufficient decomposition of nutrients, and damage to the soil. Ecological structure, leading to deterioration of microbial flooding system, decreased enzyme activity, and increased pathogenic bacteria
Leading to the decline of soil fertility, and then forced to increase the use of chemical fertilizers, forming a vicious circle
Not only increased the cost of agricultural production, but also caused a great waste of resources, and exacerbated a series of environmental problems

Method used

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  • Salt-tolerant arthrobacter and nitrogen fixation application thereof
  • Salt-tolerant arthrobacter and nitrogen fixation application thereof
  • Salt-tolerant arthrobacter and nitrogen fixation application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1: Isolation and purification of Arthrobacter halotolerant strains

[0059] 1. Culture medium preparation

[0060] (1) Enrichment medium:

[0061] Ashby nitrogen-free medium: mannitol 10g, KH 2 PO 4 0.2g, MgSO 4 ·7H 2 O 0.2g, NaCl 0.2g, CaSO 4 2H 2 O 0.1g, CaCO 3 5.0g, distilled water 1.0L;

[0062] Ashby nitrogen-free medium, sterilized at 121°C for 20 minutes;

[0063] (2) Separation and purification medium: solid Ashby nitrogen-free medium;

[0064] The solid Ashby nitrogen-free medium is prepared as follows: mannitol 10g, KH 2 PO 4 0.2g, MgSO 4 ·7H 2 O0.2g, NaCl 0.2g, CaSO 4 2H 2 O 0.1g, CaCO 3 5.0g, 15g agar, 1.0L distilled water; sterilize at 121°C for 20 minutes;

[0065] 2. Enrichment culture

[0066] The sampling site is located in Bagezhuang Village, Lucheng Town, Tongzhou District, Beijing (39°52'50.365"N, 116°46'25.986"E). The soil type is mainly clay-moist loess, and the main crop is wheat;

[0067] The collection of soil samp...

Embodiment 2

[0075] Example 2: Research on the microbiological characteristics of Arthrobacter halotolerant strains

[0076] 1. Colony morphology observation

[0077] Streak inoculation of the isolated and purified Arthrobacter halotolerant strain on Ashby nitrogen-free medium, culture at 35°C until colonies grow, and observe the colony morphology, the results are as follows:

[0078] Gram-positive, the colony is round, smooth, transparent, with neat edges, long rod-shaped bacteria, no flagella, single, and the size is about (0.8μm-1.2μm)×(0.2μm-0.4μm);

[0079] 2. Observation of cell morphology

[0080]The colonies of strains cultured on Ashby nitrogen-free medium for 2 days were picked, fixed with glutaraldehyde, rinsed with phosphate buffer (pH=7.2), dehydrated with gradient ethanol, treated with isoamyl acetate, dried at the critical point of carbon dioxide, sprayed Gold, put the sample into the observation room and carry out electron microscope scanning (German Zeiss sigma300, EHT=1...

Embodiment 3

[0090] 1. Preparation of wet cells of Arthrobacter halotolerant strain

[0091] Use an inoculation loop to pick an appropriate amount of salt-tolerant Arthrobacter colonies on the slant medium used for the preservation of strains, streak on the sterilized solid Ashby nitrogen-free medium, and cultivate at 35°C for 3 days to obtain long-term There are medium plates with obvious colonies to obtain Arthrobacter halotolerant wet cells; the solid Ashby nitrogen-free medium is prepared as follows: mannitol 10g, KH 2 PO 4 0.2g, MgSO 4 ·7H 2 O 0.2g, NaCl 0.2g, CaSO 4 2H 2 O 0.1g, CaCO 3 5.0g, 15g agar, 1.0L distilled water; sterilize at 121°C for 20 minutes.

[0092] 2. Nitrogen fixation test in medium

[0093] The wet cells of Arthrobacter halotolerant obtained in step 1 were inoculated into the enrichment medium, wherein 0.5 g of Arthrobacter halotolerant wet cells were inoculated in every 50 mL of enrichment medium; then at 35° C., pH=7.4, shaking Cultivate under the cond...

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Abstract

The invention relates to a salt-tolerant arthrobacter strain and an application thereof, and belongs to the technical field of microbiology and environmental protection. The salt-tolerant arthrobacter strain is classified and named as the salt-tolerant Arthrobacter pascens, with preservation number of CGMCC No.20974, preserved at the China General Microbiological Culture Collection Center (CGMCC) on October 29, 2020. The arthrobacter disclosed by the invention is strong and lasting in nitrogen fixation capacity, can improve content of Kjeldahl nitrogen in an Ashby nitrogen-free culture medium, and increase content of available nitrogen in soil by 15.6%. The arthrobacter can be used in nitrogen-deficient soil for converting free nitrogen elements in the air and other forms of nitrogen elements in the soil into available nitrogen, so as to improve soil fertility. The arthrobacter can be applied to soil with excessive chemical fertilizer, so as to convert other forms of nitrogen into easy-to-use available nitrogen, thereby performing biological remediation on the soil, and reducing consumption of chemical fertilizers. The arthrobacter can be prepared into a microbial fertilizer as a deep strengthening treatment approach for improving the soil fertility. The method is simple, and lower in operation and maintenance cost, and has good application prospect in the development of green agriculture.

Description

technical field [0001] The invention relates to a halotolerant Arthrobacter and its nitrogen fixation application, belonging to the technical fields of microbiology and environmental protection. Background technique [0002] Nitrogen is one of the most important nutrients for plant growth. At present, my country's agriculture wants to increase nitrogen content, mainly through the application of chemical fertilizers. However, due to lagging agricultural technology and improper fertilizer application methods, the utilization rate of chemical fertilizers is low. In rainy areas or areas with frequent irrigation, available nitrogen will be lost with rainwater or irrigation water. Not only is the loss rate extremely high, but the inflow of a large amount of nitrogen-containing sewage into the water body is also a major cause of eutrophication. [0003] Due to the excessive application of chemical fertilizers, it will lead to soil pollution, accumulation of toxic substances, chan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C05F11/08C12R1/06
CPCC12N1/20C05F11/08
Inventor 王永京满涛任连海王攀张明露王松涛闫新宇
Owner BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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