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Salt-resistant Serratiasp. strain and application thereof in restoration of salinized petroleum-contaminated soil

A technology of Serratia and Serratia, applied in the field of microbial application technology and contaminated soil bioremediation

Inactive Publication Date: 2012-03-21
BINZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are few reports on salt-tolerant petroleum-degrading microorganisms

Method used

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  • Salt-resistant Serratiasp. strain and application thereof in restoration of salinized petroleum-contaminated soil
  • Salt-resistant Serratiasp. strain and application thereof in restoration of salinized petroleum-contaminated soil
  • Salt-resistant Serratiasp. strain and application thereof in restoration of salinized petroleum-contaminated soil

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Isolation and Screening of Serratia sp. BF40 Strain

[0023] Take 10g of petroleum-contaminated coastal salinized soil, the soluble salt content in the soil is 1.2%, put the soil into a triangular flask filled with 100mL sterile water, shake at high speed for 20min, take 10ml and inoculate it into 1.0%NaCl inorganic salt solution In wax medium, medium composition: Na 2 HPO 4 1.5 g / L; KH 2 PO 4 3.84 g / L; MgSO 4 0.7g / L; (NH4) 2 SO 4 4.0 g / L; yeast powder 0.01g / L; liquid paraffin 2% (v / w); cultured on a shaker at 37°C for 5 days, repeated enrichment 3 times, fully mixed the enriched culture solution, and took 100 μL Spread on LB solid plates with 1.0% NaCl, incubate in a constant temperature incubator at 37°C for 36 hours, and observe the grown colonies. Inoculate the growing single colony into LB medium and culture to OD 630 It is about 0.8, and it is inserted into 100mL, 1.0% NaCl inorganic salt liquid wax medium as a seed solution at a ratio of 10% (v / v), an...

Embodiment 2

[0026] Determination of 16S rDNA Gene Sequence of Serratia sp. BF40 Strain

[0027] The strain was inoculated in LB medium, cultured in a constant temperature incubator at 37°C for 20 hours, collected by centrifugation, resuspended, added lysozyme and SDS to break the wall, extracted genomic DNA by phenol-chloroform method, sequenced, 16S rDNA gene sequence The length is 1503bp, and the sequence has greater than 98% similarity with the 16S rDNA sequences of different species in the genus Serratia given in the NCBI database, and its 16S rDNA gene sequences are shown in the sequence table.

Embodiment 3

[0029] Salt Tolerance of Serratia sp. BF40 Strain

[0030] Use inorganic salt medium, add 0.1% yeast powder, set five NaCl treatments with different concentration gradients: 0, 0.5%, 2.5%, 5.0%, 7.5%, after inoculation, culture in a constant temperature incubator at 37°C for 48h, measure The absorbance value of the bacterial solution, the results are shown in figure 1 .

[0031] Depend on figure 1 It can be seen that under the condition of 0-2.5% salinity, the strain can grow normally, and when the salinity increases to 5.0%, the growth amount is about half of the maximum growth amount, and the growth of the strain is significantly affected by the increase of salinity. Inhibition, but still able to grow in an environment with a salt concentration of 7%. Therefore, Serratia sp. BF40 is a salt-tolerant bacterium, and its salt-tolerant degree is 0-7% in liquid medium.

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Abstract

The invention belongs to the fields of microbial application technology and biological restoration of contaminated soil, and particularly relates to a Serratiasp. strain and application thereof in the restoration of salinized petroleum-contaminated soil. The Serratiasp. strain disclosed by the invention was preserved by China General Microbiological Culture Collection Center on September 9th, 2011, and the preservation number is CGMCC NO.5236; and the strain can efficiently degrade petroleum in a high-salt liquid environment and salinized soil. The strain is separated out from salinized petroleum-contaminated soil under salt stress conditions. The strain has regular edges, and is round in colony, non-transparent and white in color; and the thallus is a short rhabditiform and gram negativeone having a small size. The invention also discloses a nucleotide sequence of the Serratiasp. strain. The strain can degrade petroleum in a high-salt liquid environment, has a petroleum degradation rate of 60% or above in salinized soil, and has an obvious application effect in the restoration of salinized soil in the field.

Description

technical field [0001] The invention belongs to the field of microbial application technology and contaminated soil bioremediation, and in particular relates to a Serratia sp. and its application in oil-polluted salinized soil remediation. Background technique [0002] During the production and use of petroleum and its petroleum products, soil pollution problems are inevitable. After the soil is polluted by oil, the productivity will decrease, and the harmful components in the oil can also enter the water body, the atmosphere, and even agricultural products, seriously affecting the environmental quality and endangering human health. Experts from various countries have been conducting research and development and tackling key problems in the field of oil-contaminated soil remediation technology. However, due to the complexity of oil components, once it pollutes the environment, it is more difficult to restore. Bioremediation is a very potential method for the treatment of oi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20B09C1/10C12R1/425
Inventor 谢文军吴涛李小彬王君张丽燕郝雅丽
Owner BINZHOU UNIV
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