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A kind of rhizobium pusense F3-1 and its application

A technology of microbial strains and rhizobia is applied in the field of microorganisms to achieve significant economic and social benefits, reduce reproduction opportunities, and reduce the probability of crop disease.

Active Publication Date: 2021-09-21
HENAN ACAD OF SCI INST OF BIOLOGY LIABILITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] For above-mentioned situation, in order to solve the defective of prior art, the purpose of the present invention is exactly to provide a kind of Rhizobium pusense F3-1 and its application can effectively solve the problem of microbial remediation of polycyclic aromatic hydrocarbons under conditions of acidic and alkaline environments, lack of nitrogen sources and heavy metal pollution

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0010] Example 1 Screening and identification of strains

[0011] 1. Preliminary screening

[0012] Contaminated soil from the oilfield of Nanyang City, Henan Province was taken, the soil sample was passed through a 2 mm sieve, 10.00 g was accurately weighed, added to 50 mL of M9 liquid medium, placed in a shaker at 25 °C, 180 rpm for 24 h, and 5 mL of the upper medium was taken. Transferred to 45 mL M9 liquid medium containing 250 mg L -1 phenanthrene and pyrene, when the growth of microorganisms is obvious, take 5 mL of culture medium again and transfer it to fresh M9 medium containing phenanthrene and pyrene. Coat M9 solid medium containing 100 mg L -1 phenanthrene and pyrene were cultured in a 37°C incubator. After the colony has grown, pick the bacterial plate on the medium and streak to obtain a single bacterial colony. A total of 14 strains of bacteria were screened from the contaminated soil through multiple enrichment cultures and isolation and purification. These...

Embodiment 2

[0025] Example 2 Metal resistance test of strain F3-1

[0026] Investigate different concentrations of Cr 6+ and Cu 2+ Effects on strain growth, 10 g L were prepared with deionized water -1 Cr 6+ and 1g L -1 Cu 2+mother liquor. Pick a single colony of F3-1 and inoculate it in OMM medium, add 0.5% glucose as the sole carbon source in the medium, place it on a shaker for 16-20 h, and then transfer 1 mL into sterilized 6 mL OMM for culture. The medium contains 0.5% glucose and various concentrations of Cr 6+ or Cu 2+ , three replicates for each treatment, and then placed on a shaker at 25°C and 180rpm for culture, sampling at different time periods, and using a 96-well plate to detect the growth of the strain at OD600.

[0027] The selected metal ion Cr in this test 6+ Concentrations are 0, 1, 3, 5, 7, 10, 20, 50, 100 mg L -1 , the results showed that the strain was in the range of 0-50 mg L -1 can grow, when Cr 6+ Concentration of 100 mg L -1 , the strain hardly gre...

Embodiment 3

[0028] Example 3 Strain R. pusense Degradation of polycyclic aromatic hydrocarbons by F3-1 under various adverse environmental and heavy metal conditions

[0029] 1. Strain R. pusense Degradation of phenanthrene by F3-1 under various adverse environmental and heavy metal conditions

[0030] Preparation of bacterial liquid: Pick strain F3-1 from M9 solid medium, inoculate it in LB liquid medium, cultivate at 180 rpm, 37 °C, shaker for 16-24 h, then centrifuge and discard the supernatant, add 0.5% The M9 liquid medium of glucose makes the OD600 of the strain 1.0 for use. The M9 liquid medium used Ca 3 (PO 4 ) 2 Substitute NH in M9 liquid medium 4 Cl, while adding metal ion Cr 6+ and Cu 2+ to a final concentration of 20 - 40 mg L, respectively -1 , ， pH5.0-7.4.

[0031] Prepare phenanthrene standard with n-hexane at a concentration of 150 mg L -1 , add 1 mL of phenanthrene standard solution in a brown glass bottle, and take the OD after the n-hexane is completely ev...

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Abstract

The present invention relates to a Rhizobium pusense F3‑1 and its application can effectively solve the problem of microbial remediation of polycyclic aromatic hydrocarbons under the conditions of acidic and alkaline environments, lack of nitrogen sources and heavy metal pollution; the technical solution is, a Rhizobium pusense F3‑1, taxonomically named Rhizobium pusense , on August 22, 2018, was deposited in the General Microorganism Center of China Microbiological Culture Collection Management Committee, the preservation number is CGMCC NO.16332, and the address of the preservation unit is: No. 3, No. 1, Beichen West Road, Chaoyang District, Beijing; Rhizobium F3‑1 , Application in the degradation of polycyclic aromatic hydrocarbons under the conditions of acidic, alkaline environment, lack of nitrogen source and heavy metal pollution.

Description

technical field [0001] The present invention relates to the field of microorganisms, in particular to a Rhizobium pusense F3-1 and its applications. Background technique [0002] Polycyclic aromatic hydrocarbons (PAHs) are a wide range of organic compounds. Many PAHs will accumulate in organisms and cause carcinogenic, teratogenic and mutagenic effects on organisms through cytotoxicity, genotoxicity and immunotoxicity, posing a huge threat to natural biosecurity and human health. In the 1980s, the US Environmental Protection Agency included 16 PAHs without branches in the list of priority pollutants, and my country also included PAHs in the blacklist of environmental pollutants. Heavy metals (HMs), as typical inorganic pollutants, cannot be degraded, but can only be transformed. Both are easily adsorbed by soil particles, and can exist in the soil and water environment for a long time. Ecotoxicity, both of which are transferred and enriched through the food chain, ultima...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20B09C1/10C02F3/34C12R1/41C02F101/32
CPCB09C1/10C02F3/34C02F2101/327C12N1/205C12R2001/41
Inventor 甄静李冠杰王继雯杜志敏陈国参岳丹丹杨文玲赵俊杰巩涛
Owner HENAN ACAD OF SCI INST OF BIOLOGY LIABILITY
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