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Hydrogen-oxidizing bacteria A06 with growth-promoting effect and separating method and application thereof

A technology of action, separation and purification, which is applied in the field of hydrogen oxidizing bacteria, greedy phage A06 and its microbial inoculants, to achieve the effects of reducing the amount of chemical fertilizers, stabilizing crop yields, and promoting crop yields

Inactive Publication Date: 2019-08-02
NORTHWEST UNIV(CN)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Research on the isolation, cultivation, classification and identification of hydrogen oxidizing bacteria and its correlation with plant growth is only carried out in a few laboratories in the world, and domestic research in this area is still blank

Method used

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  • Hydrogen-oxidizing bacteria A06 with growth-promoting effect and separating method and application thereof
  • Hydrogen-oxidizing bacteria A06 with growth-promoting effect and separating method and application thereof
  • Hydrogen-oxidizing bacteria A06 with growth-promoting effect and separating method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] The isolation and cultivation steps of bacterial strain hydrogen oxidizing bacteria A06 of the present invention are as follows:

[0052] (1) Sample collection: Hup in Sanyuan County, Guanzhong District, Shaanxi Province was collected by conventional methods. - Soybean plant rhizosphere soil.

[0053] (2) Enrichment culture: Take 1 gram of the enriched mixed soil, put it into a triangular flask containing 100 mL of sterile water and glass beads, shake it for 20 minutes, so that the soil sample and water are fully mixed, and the cells are broken. Dispersion, serial dilution 10 -6 ∽10 -12 Coat MSA plates. Sprinkle sterilized soil (0.05 stay flat plate) on MSA then, to increase the growth attachment area of ​​hydrogen oxidizing bacteria, be beneficial to the growth of hydrogen oxidizing bacteria. Put the plate into an airtight container with a flow rate of 280 mL min -1 , containing H 2 The amount is 1.25×10 -4 mol L -1 Mixed gas, invert culture for 2∽4 weeks, unti...

Embodiment 2

[0071] Embodiment 2 The utilization of hydrogen oxidizing bacteria A06 to different carbon sources

[0072] Due to the limitation of the volume of the container for the cultivation of hydrogen oxidizing bacteria in the hydrogen cultivation device, the number of cultures should not be too large, and the selected hydrogen oxidizing bacteria have the ability to grow in organic carbon source medium. In order to further study other physiological characteristics of hydrogen-oxidizing bacteria strains that promote wheat root growth and the possible mechanism of promoting wheat root growth, different carbon sources were added to the MSA screening medium to investigate the growth of hydrogen-oxidizing bacteria under different carbon source conditions. growing situation.

[0073] Add glucose, maltose, soluble starch, xylose, fructose, and sucrose to the MSA liquid medium, and add six commonly used carbon sources. After inoculating the bacteria, culture them on a shaker at 37°C with a sp...

Embodiment 3

[0076] Embodiment 3 has the detection of the hydrogen oxidizing bacteria of ACC deaminase

[0077] 1. Detection of hydrogen oxidizing bacteria with ACC deaminase

[0078] The hydrogen oxidizing strains that can promote the significant growth of wheat roots in the wheat plate test were taken to detect the activity of ACC deaminase. The experimental steps are as follows:

[0079] (1) The detection conditions determined that ACC is a non-protein amino acid, so it was detected by amino acid chromatography. A mixed solution of n-butanol, acetic acid, and water in different proportions was prepared as a spreading agent, a silica gel plate was used as a support, and a chromogenic agent was 0.5% ninhydrin solution, and the optimal spreading agent ratio was tested.

[0080] (2) ACC liquid medium The MSA medium was simply changed, and the agar was removed from the medium, that is, the solid MSA medium was changed into a liquid MSA medium. At the same time, soluble starch was added as...

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Abstract

The invention discloses variovorax paradoxus with a growth-promoting effect. The variovorax paradoxus with the growth-promoting effect is classified and named Variovorax paradoxus A06, the Variovoraxparadoxus A06 is preserved at the China Center for Type Culture Collection on 22th, October, 2018, and a preservation number is CCTCC No: M 2018696. According to the variovorax paradoxus with the growth-promoting effect, leguminosae rhizosphere soil is used as a study object, and a continuous hydrogen introduction device is used for conducting isolated culture on hydrogen-oxidizing bacteria; and by conducting morphologic observation, physiology and biochemistry study and 16SrDNA identification, a taxonomic status of the hydrogen-oxidizing bacteria is determined, and gas chromatography is usedfor detecting the hydrogen-oxidizing capability of the strain. The variovorax paradoxus has growth-promoting effects on wheat and corn, and the biological preparation can promote plant growth and improve the yield, the fertilizer efficacy and the anti-adversity ability of plants when used for agricultural production.

Description

technical field [0001] The invention relates to a hydrogen oxidizing bacterium Variovorax paradoxus A06 isolated and cultured from the rhizosphere soil of leguminous plants without hydrogen-absorbing enzyme and its microbial agent. The hydrogen oxidizing bacteria A06 strain was deposited in the Chinese Center for Typical Cultures of Wuhan University on October 22, 2018, and the preservation number is CCTCC M 2018696. The bacterium can promote plant growth and belongs to the technical field of microorganisms. Background technique [0002] The development of urbanization, the rapid growth of economy, the rapid expansion of population and the rapid reduction of arable land make agricultural production face enormous pressure. In order to maintain the growth of agricultural output, chemical fertilizers are applied in a large number of places to obtain short-term benefits. However, the long-term use of a large amount of chemical fertilizers has caused the compaction and degradat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02A01N63/00A01P21/00C12R1/01
CPCA01N63/00C12N1/02C12N1/20C12N1/205C12R2001/01Y02A40/10
Inventor 王卫卫李璐璐刘瑞瑞李志英
Owner NORTHWEST UNIV(CN)
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