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A siderophore-producing hydrogen-oxidizing bacterium and its isolation method and application

A technology of hydrogen oxidation and bacteria, applied in microorganism-based methods, biochemical equipment and methods, immobilized on or in inorganic carriers, etc., can solve problems such as few reports

Active Publication Date: 2022-05-17
西安绿海生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, domestic research on hydrogen oxidizing bacteria has not been in-depth, and there are few related reports.

Method used

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  • A siderophore-producing hydrogen-oxidizing bacterium and its isolation method and application
  • A siderophore-producing hydrogen-oxidizing bacterium and its isolation method and application
  • A siderophore-producing hydrogen-oxidizing bacterium and its isolation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The separation and screening of embodiment 1 soil microorganism

[0041] The strain was isolated from soybean fields in Bailiang Village, Bailiangzhai Village and Leijia Village, Shuangzhao Town, Xianyang City, Shaanxi Province. Weigh 5g of the rhizosphere soil sample, put into a Erlenmeyer flask with small glass beads and 45mL of sterile water to make 10 -1 Soil suspension, put the soil suspension into a shaker at 180r / min for 30min. Dilute the shaken soil suspension to 10 -11 , and then spread different concentrations of soil suspension on the mineral salt medium, put it into the gas circulation system and cultivate it upside down for 3-4 weeks. Three replicates were set up for each concentration. Colonies of different sizes and shapes were selected from the plate to be streaked and purified on the mineral salt medium, and the purified strains were inoculated on the slope of beef extract and stored in a 4°C refrigerator.

[0042] Inoculate the strain to be tested ...

Embodiment 2

[0045] Example 2 Determination of strain secretion IAA ability

[0046] The hydrogen oxidizing bacteria strain that will be screened, after being activated on the beef extract peptone medium, inoculate in the KingB-Trp medium, wherein the medium contains the L-Trp that final concentration is 100mg / L, then in the shaker 28 Cultivate with shaking at 180r / min for 3 days. Place the culture solution in a centrifuge at 5000r / min for 20min, discard the precipitate, take 0.75mL of the supernatant and mix it with 0.75mL of Salkowski Reagent evenly, react in the dark for 20min, observe the color of the reaction solution, if the reaction solution is pink , the strain can secrete IAA. The OD value of the reaction liquid was measured at 530nm, and the reaction liquid of the sterilized medium and Salkowski Reagent was used as the control group. Substitute the measured OD value into the standard curve to calculate the concentration of IAA produced. In order to ensure the accuracy of the e...

Embodiment 3

[0049] Embodiment 3 bacterial strain siderophore assay

[0050] The determination of siderophores produced by strains is a double-layer CAS plate culture method, and the specific steps are as follows: inoculate the screened hydrogen-oxidizing bacterial strains into LB medium, place them in a shaker at 30°C, and cultivate them at 200r / min for 24 hours. The strains were inoculated into the iron-free MKB liquid medium, placed in a shaker at 30°C, 200r / min iron starvation treatment for 72h, then the strains were inoculated into the iron-free MKB solid medium, placed in a biochemical incubator at 30°C and inverted For culture, add the sterilized upper medium to the lower medium with obvious colonies, and observe the color change around the strains. If orange halos appear around the colonies, siderophores are produced. Otherwise, nothing. The greater the ability of the strain to secrete siderophores, the larger the halo, the darker the color, and the shorter the time of appearance....

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Abstract

The invention discloses a Arthrobacter genus Arthrobacter 201‑9, which is isolated from the soybean fields of Bailiang Village, Bailiangzhai Village and Leijia Village, Shuangzhao Town, Xianyang City, Shaanxi Province, and is classified as Arthrobacter based on physiological and biochemical characteristics and 16SrDNA analysis. Bacteria. The present invention takes the soybean rhizosphere soil as the research object, utilizes the gas circulation culture system to separate the hydrogen oxidizing bacteria therefrom, utilizes gas chromatography to detect whether the strains obtained contain hydrogen-absorbing enzymes, and conducts morphological characteristics, physiological Biochemical characteristics and 16SrDNA identification, combined with the above experimental results to determine the species of the strain. To study the growth-promoting characteristics of the screened hydrogen-absorbing-positive strains such as IAA and siderophore, to understand the growth-promoting mechanism of hydrogen-oxidizing bacteria, and to provide a theoretical basis for the "hydrogen fertilizer" theory.

Description

technical field [0001] The present invention relates to a hydrogen oxidizing bacterium 201-9 and its isolation method and application, in particular to a hydrogen oxidizing bacterium Arthrobacter ( Arthrobacter sp.) and its microbial agents. The strain was deposited in the Chinese Center for Typical Cultures of Wuhan University on September 19, 2018, with the preservation number CCTCC M 2018637. The bacterium can promote plant growth and belongs to the technical field of microorganisms. Background technique [0002] Crop rotation and intercropping are two widely used agricultural measures, which occupy an important position in agricultural cultivation methods. Crop rotation and intercropping can be traced back to the Han Dynasty more than 2,000 years ago, and it was widely used in actual agricultural production in the Wei, Jin, Southern and Northern Dynasties. Crop rotation and intercropping can improve the chemical composition and physical structure of the soil, increas...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N1/02C12N11/14C12N11/10C12R1/06
CPCC12N1/20C12N1/02C12N11/14C12N11/10C12R2001/06C12N1/205
Inventor 王卫卫李璐璐刘瑞瑞李志英
Owner 西安绿海生物科技有限公司
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