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Soybean rhizosphere nitrogen-fixing bacteria producing acc deaminase and antagonizing various pathogenic fungi and its application

A technology of pathogenic fungi and deaminase, applied in the direction of enzymes, applications, bacteria, etc., to achieve good inoculation effects, broad application prospects, and strong competitive adaptability

Inactive Publication Date: 2011-12-21
INST OF AGRI RESOURCES & REGIONAL PLANNING CHINESE ACADEMY OF AGRI SCI
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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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  • Soybean rhizosphere nitrogen-fixing bacteria producing acc deaminase and antagonizing various pathogenic fungi and its application
  • Soybean rhizosphere nitrogen-fixing bacteria producing acc deaminase and antagonizing various pathogenic fungi and its application
  • Soybean rhizosphere nitrogen-fixing bacteria producing acc deaminase and antagonizing various pathogenic fungi and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1, the isolation and identification of soybean rhizosphere nitrogen-fixing bacteria GD571

[0055] 1. Enrichment and isolation of soybean rhizosphere nitrogen-fixing bacteria

[0056] 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 cult...

Embodiment 2

[0087] Example 2, Determination of Nitrogenase Activity of Burkholderia sp. GD571 CGMCC No.5039

[0088] Carry out nitrogenase activity assay to Burkholderia sp. (Burkholderia sp.) GD571 CGMCC No.5039 obtained in embodiment 1, specific method is as follows: in 15 * 150mm screw-top glass tubes, add 5ml to improve nitrogen fixation culture The base was made into a slant, inoculated with Burkholderia sp. GD571 CGMCC No.5039, and cultured at 28°C. At the same time, 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 production with a gas chromatograph, and calculate ...

Embodiment 3

[0091] Embodiment 3, Burkholderia sp. (Burkholderia sp.) GD571 CGMCC No.5039 antagonizes pathogenic fungus bacteriostatic rate determination

[0092] The Burkholderia sp. (Burkholderia sp.) GD571CGMCC No.5039 obtained in embodiment 1 is adopted to measure the antibacterial rate of antagonizing pathogenic fungi by two-point confrontation method. Inoculate crops with Gibberella zeae (or Sclerotinia sclerotiorum, or Verticillium dahliae) and Burkholderia sp. GD571 CGMCC No .5039, each screening process was repeated 3 times, and the flat plate of only receiving pathogenic fungi but not receiving Burkholderia sp. (Burkholderia sp.) GD571 CGMCC No.5039 was used as a control. Incubate at a constant temperature at 28°C, and measure the colony radius r of the pathogenic fungi on the confrontation plate along the direction of the tested Burkholderia sp. GD571 CGMCC No.5039 with a millimeter scale after 15 days 1 , and the colony radius r of pathogenic fungi on the control plate 0 . G...

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Abstract

The invention discloses a soybean rhizosphere azotobacter for producing activated calcium carbonate (ACC) and realizing the antagonistic effect on various pathogenic fungi and a purpose thereof. The soybean rhizosphere azotobacter is Burkholderia sp. GDSD571 and has the preservation number being CGMCC No.5039 in General Microbiology Center of China Microbial Strain Preservation Management Committee. The Burkholderia sp. GDSD571 CGMCC No.5039 provided by the invention has high nitrogen fixation activity and ACC ammonialyase activity, can realize the effective antagonistic effect on gibberella zeae, sclerotinia sclerotiorum and verticillium dahliae and has wide application prospects in nitrogen fixation microbial agent and microbial organic fertilizer production.

Description

technical field [0001] The invention relates to soybean rhizosphere nitrogen-fixing bacteria that produce ACC deaminase and antagonize various pathogenic fungi and uses thereof, in particular to a kind that can produce ACC deaminase, antagonize crop scab, sclerotinia and verticillium dahliae, and Bacteria capable of efficient nitrogen fixation in the 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 ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/88C12N9/00A01N63/02A01P3/00A01P21/00C05F11/08C12R1/01
Inventor 孙建光徐晶高淼胡海燕
Owner INST OF AGRI RESOURCES & REGIONAL PLANNING CHINESE ACADEMY OF AGRI SCI
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