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Brevibacterium halotolerans and application thereof in ecological restoration of soils

A technology of Brevibacterium salt-tolerant and inoculum, which is applied in microorganism-based methods, bacteria, microorganisms, etc., to achieve the effect of improving soil fertility, broad application prospects, and soil improvement

Active Publication Date: 2017-02-22
北京复天科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the bottleneck restricting the development of my country's microbial fertilizer industry is the selection of high-efficiency strains

Method used

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  • Brevibacterium halotolerans and application thereof in ecological restoration of soils
  • Brevibacterium halotolerans and application thereof in ecological restoration of soils

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1. Isolation and Identification of Brevibacterium halotolerans GDSD21 CGMCC No.12402

[0050] 1. Enrichment and isolation of crop rhizosphere nitrogen-fixing bacteria GDSD21

[0051] Take 10g of soil sample (collected from Suihua paddy field in Heilongjiang Province, China) and put it into 90mL of sterile water and shake it for 20min to make a cloudy solution, draw 5mL into 30mL of nitrogen-fixing bacteria enrichment culture medium ACCC55, shake at 100rpm, 28℃ Shaking culture, after 72h, replace with fresh medium to continue the culture. The nitrogen-fixing bacteria were isolated after repeating the culture for 3 times. Draw 1mL of the above nitrogen-fixing bacteria enrichment culture into 9mL sterile water to make 10 -1 Dilution, continue to dilute to make 10 -2 、10 -3 、10 -4 、10 -5 Take 0.1mL of each dilution of the bacterial suspension and spread it on a nitrogen-free solid medium plate, and culture it statically at 29°C. After 2-3 days of colony forma...

Embodiment 2

[0076] Example 2, Nitrogenase Activity Determination of Brevibacterium halotolerans GDSD21 CGMCC No.12402

[0077] The nitrogenase activity measurement was carried out as the experimental group with the commonly used microbial fertilizer Azotobacter rotoleans ACCC11103 as the control group, and the Brevibacterium halotolerans (Brevibacterium halotolerans) GDSD21 CGMCC No.12402 of Example 1. The specific method is as follows: Add 5 mL of modified nitrogen-fixing medium into a screw-top glass tube to make a slant, inoculate the strain, and culture at 28°C. The blank slant without inoculation was used as the negative control, and three replicates were set up for each treatment. After culturing for 72 hours, replace the rubber stopper, inject acetylene gas to make the final concentration 10%, seal it with medical adhesive tape, continue culturing for 72 hours, take 100 μL of reaction gas, measure the amount of ethylene produced by gas chromatography, and calculate the nitrogenase ...

Embodiment 3

[0080] Example 3, application of Brevibacterium halotolerans GDSD21 CGMCC No.12402 in ecological restoration

[0081] 1. Preparation of bacterial agents for soil ecological restoration

[0082] Utilize the Brevibacterium halotolerans (Brevibacterium halotolerans) GDSD21CGMCC No.12402 obtained in Example 1 to prepare the soil ecological restoration bacterial agent, the specific method is as follows: adopt the above-mentioned improved nitrogen-fixing liquid medium, and cultivate the salt-tolerant Brevibacterium halotolerans on a shaker under laboratory conditions. Bacillus (Brevibacterium halotolerans) GDSD21 CGMCCNo.12402. 1L Erlenmeyer bottle capacity 250mL, 28°C, 200rpm shaking culture for 18h ~ 24h to OD 600 The content of Brevibacterium halotolerans GDSD21 CGMCC No.12402 reached 1.5-1.8, and the content of Brevibacterium halotolerans GDSD21 CGMCC No.12402 reached 1.5-2 billion cfu / mL. Canned and sealed to obtain the soil ecological restoration bacterial agent. Store at ro...

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Abstract

The invention discloses a brevibacterium halotolerans with relatively high azotase activity and application thereof in ecological restoration. The brevibacterium halotolerans GDSD21 is collected in the China General Microbiological Culture Collection Center with the collection number of CGMCC No.12402. Nitrogen-fixing bacteria are separated from rhizosphere soils of field-crop roots, further strains with relatively high azotase activity are screened, and finally the efficient nitrogen-fixing bacterium brevibacterium halotolerans GDSD21 is screened. The azotase activity of the brevibacterium halotolerans GDSD21 is obviously higher than that of common production strain azotobacter chroococcum ACCC11103 for microbial fertilizers, and the brevibacterium halotolerans GDSD21 has extensive application prospects in soil improvement, increasing of the soil fertility, ecological restoration of a virgin land, and production of industrialized seedling-rearing inoculation agents, nitrogen-fixing microbial agents and bio-organic fertilizers for vegetables.

Description

technical field [0001] The invention relates to a strain of Brevibacterium halotolerant with higher nitrogenase activity and its application in ecological restoration. Background technique [0002] Nitrogen is an essential chemical element for plant growth, and nitrogen fertilizer is one of the most important means of production in agricultural ecological restoration and agricultural production. The production of chemical nitrogen fertilizer consumes a large amount of energy, accounting for 70% to 80% of the total production cost. Fossil energy such as coal, oil, and natural gas is non-renewable, and agricultural production that relies on chemical nitrogen fertilizers is difficult to sustain. Chemical nitrogen fertilizers have played a huge role in increasing crop yields and ensuring food security in my country, but unreasonable nitrogen application has caused serious environmental pollution, causing a series of serious hazards, such as cyanobacteria pollution in Taihu Lake...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C05F11/08C12R1/13
CPCC05F11/08C12N1/20C12N1/205C12R2001/13
Inventor 张春禹范天立赵平孙建光
Owner 北京复天科技有限公司
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