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Bacillus derived from soybean rhizosphere and capable of antagonizing pathogenic fungi and application thereof

A technology of bacillus and inoculum, which is applied in the field of soybean rhizosphere nitrogen-fixing bacteria to achieve broad application prospects, strong competitive adaptability and good inoculation effect.

Inactive Publication Date: 2013-07-31
哈尔滨君龙实业有限公司 +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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  • Bacillus derived from soybean rhizosphere and capable of antagonizing pathogenic fungi and application thereof
  • Bacillus derived from soybean rhizosphere and capable of antagonizing pathogenic fungi and application thereof
  • Bacillus derived from soybean rhizosphere and capable of antagonizing pathogenic fungi and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1, the isolation and identification of soybean rhizosphere nitrogen-fixing bacteria GD5711

[0043] 1. Isolation of soybean rhizosphere nitrogen-fixing bacteria GD5711

[0044] Take 10g soil sample (collected from Heilongjiang, China) and shake it in 90ml sterile water for 20min to make a turbid solution, absorb 5ml and put it into 30ml nitrogen-fixing bacteria enrichment culture medium ACCC55, 100rpm, 28℃ for shaking culture, 72h Then replace with fresh medium to continue culturing. 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 was spread on a nitrogen-fixing bacteria isolation medium plate, and cultured at...

Embodiment 2

[0073] Embodiment 2, determination of nitrogenase activity of Bacillus sp. GD5711CGMCC No.5152

[0074] The Bacillus sp. (Bacillus sp.) GD5711CGMCC No.5152 obtained in Example 1 is carried out nitrogenase activity assay, and specific method is as follows: in 15 * 150mm screw-top glass tubes, add 5ml improved nitrogen fixation medium to make inclined-plane, inoculate Azotobacter, cultivated at 28°C. A blank slant that was not inoculated was used as a negative control, and three replicates were set up. 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 amount of Bacillus sp .) Nitrogenase activity of GD5711CGMCC No.5152. Nitrogenase activity (nmol / mg h) = C 2 h 4 nmol / [thalline protein amount (mg) × reaction time (h)], where C 2 h 4 ...

Embodiment 3

[0077] Embodiment 3, Bacillus sp. (Bacillus sp.) GD5711CGMCC No.5152 antagonizes pathogenic fungus inhibition rate determination

[0078] The Bacillus sp. GD5711CGMCC No.5152 obtained in Example 1 is adopted to measure the bacteriostatic rate of antagonizing pathogenic fungi by two-point confrontation method. Inoculate crops with Gibberella zeae (or Sclerotinia sclerotiorum) and Bacillus sp. GD5711CGMCC No.5152, each screening treatment has 3 replicates, so that only pathogenic fungi and no nitrogen-fixing bacteria are inoculated on the plate As a contrast. 28 ℃ constant temperature culture, after 15 days, measure the colony radius r of the pathogenic fungus on the confrontation plate along the direction of the tested Bacillus (Bacillus sp.) GD5711CGMCC No.5152 with a millimeter scale 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 culture pathogenic fungus colony rad...

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Abstract

The invention discloses soybean rhizosphere azotobacter capable of antagonizing various pathogenic fungi and application thereof. The soybean rhizosphere azotobacter is bacillus (Bacillus sp.) GD5711 and has the preservation number of CGMCC No.5152 in the China General Microbiological Culture Collection Center (CGMCC). The bacillus (Bacillus sp.) GD5711 CGMCC No.5152 provided by the invention hasnitrogen fixation activity, can effectively antagonize crop gibberella saubinetii (Gibberellazeae) and sclerotinia sclerotiorum (Sclerotinia sclerotiorum) and has wide application prospect in production of a nitrogen-fixing microorganism inoculant and a bio-organic fertilizer.

Description

technical field [0001] The invention relates to a soybean rhizosphere nitrogen-fixing bacterium that antagonizes various pathogenic fungi and uses thereof, in particular to a bacterium that can antagonize crop scab and sclerotinia and has nitrogen-fixing function in soybean rhizosphere. 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 applica...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/02A01P3/00C05F11/08C12N9/00C12R1/07
Inventor 宋燕军王会智孙建光
Owner 哈尔滨君龙实业有限公司