Sclerotinia sclerotiorum-antagonizing nitrogen-fixing spore bacterium and its application

A technology of bacillus and nitrogenase, applied in the direction of application, bacteria, fungicides, etc., to achieve good inoculation effect, broad application prospects, and high nitrogenase activity

Active Publication Date: 2013-01-16
上海国环生态科技股份有限公司 +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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  • Sclerotinia sclerotiorum-antagonizing nitrogen-fixing spore bacterium and its application
  • Sclerotinia sclerotiorum-antagonizing nitrogen-fixing spore bacterium and its application
  • Sclerotinia sclerotiorum-antagonizing nitrogen-fixing spore bacterium and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1, the separation and identification of nitrogen-fixing bacteria GDSD212

[0042] 1. Enrichment and isolation of nitrogen-fixing bacteria GDSD212

[0043] Take 10g of soil sample (farmland soil with relatively poor nitrogen nutrition in Heilongjiang Province, China) and put it into 90ml of sterile water and shake it for 20min to make a cloudy solution. Take 5ml and put it into 30ml of nitrogen-fixing bacteria enrichment culture medium ACCC55, 100rpm, 28℃ Carry out shaking culture on a shaker, and change fresh culture medium after 72h to continue the culture. The nitrogen-fixing spores were isolated after repeating the culture for 3 times. Draw 1ml of the above-mentioned nitrogen-fixing bacteria enrichment culture and put it into 9ml sterile water to make 10 -1 Dilution, continue to dilute to make 10 -2 、10 -3 、10 -4 、10 -5 Bacterial suspensions of dilutions were heated in boiling water at 100°C for 10 minutes, and after cooling, 0.1ml of each dilution w...

Embodiment 2

[0064] Embodiment 2, determination of nitrogenase activity of Bacillus sp. GDSD212 CGMCC No.5035

[0065] The Bacillus sp. (Bacillus sp.) GDSD212 CGMCC No.5035 gained in Example 1 is carried out nitrogenase activity assay, and concrete operation is as follows: in 15 * 150mm screw-top glass tubes, add 5ml improved nitrogen fixation medium to make inclined-plane, inoculate Bacillus sp. GDSD212 CGMCC No.5035, cultivated at 28°C. Azotobacter chroococcum (Azotobacter chroococcum) ACCC11103, which is commonly used in the production of microbial fertilizers, was used as the positive control, and the blank slant without inoculation was used as the negative control, with 3 replicates. 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

[0068] Embodiment 3, Bacillus sp. (Bacillus sp.) GDSD212 CGMCC No.5035 antagonizes pathogenic fungus bacteriostatic rate determination

[0069] The Bacillus sp. GDSD212 CGMCC No.5035 obtained in embodiment 1 is carried out to antagonize pathogenic fungus bacteriostatic rate measurement by two-point confrontation method. The crop pathogenic fungi Sclerotinia sclerotiorum and Bacillus sp. GDSD212 CGMCC No.5035 were inoculated respectively, and each screening treatment was repeated three times. The plate inoculated with only pathogenic fungi but not nitrogen-fixing bacteria was used as a control. Incubate at a constant temperature at 28°C, measure the colony radius r of the pathogenic fungi on the confrontation plate along the direction of the tested Bacillus sp. GDSD212 CGMCC No.5035 with a millimeter scale after 15 days 1 , and the colony radius r of pathogenic fungi on the control plate 0 . Growth inhibition rate of pathogenic fungi (%)=(control radius r 0 - confrontation c...

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Abstract

The invention discloses a Sclerotinia sclerotiorum-antagonizing nitrogen-fixing spore bacterium and its application. The nitrogen-fixing spore bacterium is Bacillus sp. GDSD212, which is preserved in General microbiological center of China Committee for Culture Collection of Microorganisms with a preservation number of CGMCC No.5035. The Bacillus sp. GDSD212 CGMCC No.5035 provided in the invention has high nitrogen-fixing activity, can effectively antagonize Sclerotinia sclerotiorum, and has a broad application prospect in production of nitrogen-fixing microorganism bacterial agents and bio-organic fertilizers.

Description

technical field [0001] The invention relates to nitrogen-fixing spores antagonizing Sclerotinia sclerotiorum and application thereof. Background technique [0002] Nitrogen fertilizer is one of the most important means of production in agricultural production. The production of chemical nitrogen fertilizer consumes a large amount of energy, accounting for 70% to 80% of the total production cost. In recent years, energy has become increasingly tense, and the price of coal for chemical fertilizer production in my country has soared, leading to an increase in the price of chemical nitrogen fertilizers, which is unbearable for farmers and has a strong response. In addition, chemical nitrogen fertilizers have played a huge role in increasing crop yields and ensuring food security in my country, but unreasonable nitrogen application has formed serious non-point source pollution, causing a series of serious hazards, such as the shocking cyanobacteria incident in Lake Tai. Nitrate...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/00A01P3/00C12N9/02C05F11/08C05G3/00C12R1/07
Inventor 周剑浩孙建光江峰徐晶张友恒
Owner 上海国环生态科技股份有限公司
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