Agrobacteriumsp. having free-living nitrogen fixing ability, and applications thereof

A technology of Agrobacterium and ability, applied in the field of microbial application, can solve the problems of inability to remediate polluted soil, low ability to resist heavy metal growth and nitrogen fixation, slow growth, etc.

Inactive Publication Date: 2012-10-24
陕西省能源化工研究院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most wild nitrogen-fixing bacteria have the disadvantages of slow growth, heavy metal resistance and low nitrogen fixation ability, which cannot meet the needs of contaminated soil remediation

Method used

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  • Agrobacteriumsp. having free-living nitrogen fixing ability, and applications thereof
  • Agrobacteriumsp. having free-living nitrogen fixing ability, and applications thereof
  • Agrobacteriumsp. having free-living nitrogen fixing ability, and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] According to the screening method of the nitrogen-fixing bacterial strain provided by the present invention, a nitrogen-fixing bacterial strain with high nitrogenase activity capable of tolerance to heavy metal cadmium is selected, and the preparation steps are as follows:

[0060] 1) Agrobacterium YSGD06, which was screened and preserved from the plant rhizosphere in the laboratory, was used as the starting strain;

[0061] 2) Mutation breeding

[0062] (1) Prepare the single cell suspension of the starting strain YSGD06

[0063]The starting strain YSGD06 was inoculated in liquid medium A, cultured at 28°C, 150rpm for 16hrs, centrifuged, washed with sterile saline, placed in a conical flask with glass beads, and oscillated to disperse into a single-cell bacterial suspension liquid;

[0064] The composition of the medium A is: yeast powder 10g, NaCl 5g, peptone 10g, FeSO 4 ·7H 2 O 0.005g, Na 2 MoO 4 2H 2 O 0.0025g, distilled water 1000ml, pH 7.0~7.2.

[0065] (...

Embodiment 2

[0076] According to the method for preparing nitrogen-fixing bacterial agent provided by the present invention, the nitrogen-fixing bacterial agent with high activity of nitrogenase that can tolerate heavy metal cadmium is prepared, and the steps of preparation are as follows:

[0077] The preparation of bacterial agent comprises the following steps:

[0078] 1. Strain fermentation

[0079] ①First-level Erlenmeyer flask liquid culture

[0080] The frozen Ymu6-2 was quickly thawed at 37°C, transferred to the Erlenmeyer flask containing liquid culture A according to the inoculum amount of 0.5%, and cultured with shaking at 150rpm at 28°C for 18hrs;

[0081] The composition of medium A is: yeast powder 10g, NaCl 5g, peptone 10g, FeSO 4 ·7H 2 O 0.01g, Na 2 MoO 4 2H 2 O 0.005 g, distilled water 1000ml, pH 7.0~7.2;

[0082] ②Secondary fermentation tank fermentation

[0083] 1) Put the primary culture medium of Ymu6-2 into a fermenter equipped with liquid medium E according ...

Embodiment 3

[0088] Use embodiment 2 bacterial agents to repair cadmium-contaminated soil in strengthened Indian mustard, and its specific steps are:

[0089] The potting soil was collected from the farmland soil in Chang’an County, Shaanxi Province. The total Cd content of the tested soil was 0.25 mg / kg, and 4 soils were added with Cd 2+ Level, respectively 0, 20, 40, 80mg / kg, the corresponding amount of CdCl 2 Dubbed into a solution, so that the test soil and heavy metals were repeatedly mixed evenly, and allowed to stand for 30 days to balance. In the experiment, a blank control group, a urea-added group and a bacterial agent-added group were respectively set up. After emergence, keep 3 plants in each pot. Dilute the liquid bacterial agent prepared above with water at a ratio of 1:200, and use the method of root irrigation to pour 20ml of the diluted bacterial liquid into each pot, once every 3 weeks, without adding bacteria The group poured the same dose of dead bacteria, and the co...

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Abstract

The present invention discloses Agrobacteriumsp. having free-living nitrogen fixing ability, wherein a classification name of the Agrobacteriumsp. is Agrobacteriumsp. Ymu6-2, the Agrobacteriumsp. is preserved in the China General Microbiological Culture Collection Center (CGMCC), and a preservation number is CGMCC No.5007. The Agrobacteriumsp. having free-living nitrogen fixing ability in the present invention has the following advantages that: high nitrogen fixing ability is provided; high concentration Cd growth can be resisted; indole-3-acetic acid can be secreted to produce iron atoms; with the prepared microbial agent, over-accumulated plant growth can be significantly promoted, biomass of over-accumulated plants can be increased, a heavy metal enrichment coefficient and a heavy metaltransfer coefficient of over-accumulated plants can be substantially increased, soil ecology environment can be improved, and advantages of environment-friendliness and low cost are provided.

Description

technical field [0001] The invention relates to an Agrobacterium strain capable of autogenous nitrogen fixation. The Agrobacterium is obtained by ultraviolet-plasma compound mutagenesis, has the ability to resist the growth of heavy metals, can be used for environmental treatment of soil heavy metal pollution, and belongs to the technical field of microbial application. Background technique [0002] Heavy metal pollution refers to environmental pollution caused by heavy metals or their compounds, mainly caused by human factors such as mining, waste gas discharge, and sewage irrigation. In recent years, with the growth of population, the rapid development of industry, and the extensive use of chemical fertilizers and pesticides, soil heavy metal pollution has become increasingly serious. [0003] Cd is a highly toxic heavy metal. The Japanese "Itai-Itai Disease" that shocked the world was caused by cadmium pollution. Cadmium will replace calcium in bone, softening and breaki...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20B09C1/10A01C1/00A01C21/00C05F11/08C12R1/01
Inventor 朱晓丽马沛范代娣申烨华梁丽华
Owner 陕西省能源化工研究院
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