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

A technology of Agrobacterium and ability, applied in the field of microbial applications, can solve the problems of slow growth, inability to meet contaminated soil remediation, anti-heavy metal growth and low nitrogen fixation capacity

Inactive Publication Date: 2013-09-18
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 method for screening nitrogen-fixing strains provided by the present invention, nitrogen-fixing strains with high nitrogenase activity that can tolerate heavy metal cadmium are selected, and the preparation steps are as follows:

[0060] 1) Take the Agrobacterium YSGD06 that the laboratory screened and preserved from the plant rhizosphere as the starting strain;

[0061] 2) Mutagenesis and breeding

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

[0063] The starting strain YSGD06 was inoculated into liquid medium A, cultured at 28°C, 150rpm for 16hrs, centrifuged, washed with sterile saline, placed in an Erlenmeyer flask filled with glass beads, shaken to make it dispersed into a single cell 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] (2) UV mutagenesis

[0066] Adjust the concent...

Embodiment 2

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

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

[0078] 1. Strain fermentation

[0079] ①First level flask liquid culture

[0080] Quickly thaw the frozen Ymu6-2 at 37°C, insert 0.5% of the inoculum into the Erlenmeyer flask containing liquid culture A, and culture with shaking at 28°C at 150rpm 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) Connect the primary culture solution of Ymu6-2 to the fermenter equipped with liquid medium E according to the inoculum of 5%, and carry out fermentation culture, the temperat...

Embodiment 3

[0088] Using the bacterial agent of Example 2 to remediate cadmium-contaminated soil with intensified Indian mustard, the specific steps are:

[0089] The potting soil was collected from farmland soil in Chang'an County, Shaanxi Province. The total Cd content of the tested soil was 0.25 mg / kg. The soil was set with 4 Cd dosings. 2+ The levels are 0, 20, 40, 80 mg / kg, and the corresponding amount of CdCl 2 Prepare the solution so that the test soil and heavy metals are repeatedly mixed evenly, and left to stand for 30 days. The experiment set up a blank control group, a urea addition group and a bacteria addition group. After emergence, keep 3 strains in each pot. Dilute the above-prepared liquid inoculum with water at a ratio of 1:200. Use the root irrigation method to pour 20ml of diluted bacteria in each pot. Pour it every 3 weeks without adding bacteria. The same dose of dead bacteria was poured in the group, and the urea concentration of the urea group was 0.12g / L. The plan...

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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 CGMCCNo.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 metal transfer 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 Patents(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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