Pseudomonas strain and screening method thereof

A technology for pseudomonas and strains, which is applied in the field of Pseudomonas strains and their screening, and can solve the problems of difficulty in maintaining the biomass of PCBs and unsatisfactory application results.

Inactive Publication Date: 2015-12-09
SHANGHAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the application results of these strains in the actual polluted soil are not ideal, the solution efficiency is only 1 / 50 of the suspension cells in the laboratory, and 1 / 2 of the soil simulation in the laboratory
Numerous studies have shown that the main reason is that these strains obtained from special locations have a special survival mode, which creates a survival competition with the indigenous microbial flora, making it difficult to maintain the biomass required for the effective degradation of PCBs in the actual soil. Become an obstacle to the reduction of non-point source pollution

Method used

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  • Pseudomonas strain and screening method thereof
  • Pseudomonas strain and screening method thereof
  • Pseudomonas strain and screening method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1, the preparation of culture medium

[0023] 1) Synthetic medium:

[0024] Solution A: 56.77g·L -1 K 2 HPO 4 , 21.94g·L -1 K H 2 PO 4 , 12.96g·L -1 (NH 4 ) 2 SO 4 ;

[0025] Solution B: 0.3g·L -1 CaCl 2 ·H 2 O, 19.5g L -1 MgSO 4 , 5g·L -1 MnSO 4 ·H 2 O, 1g L -1 FeSO 4 ·7H 2 O;

[0026] Solution C: 0.1g·L -1 Yeast extract.

[0027] Liquid synthetic medium (LSM): 77.5mL A solution + 10mL B solution + 910mL LC solution + 2g biphenyl, pH7.2.

[0028] Solid synthetic medium (SSM): 77.5mL A solution + 10mL B solution + 910mL LC solution + 2g biphenyl + 15g agar, pH7.2.

[0029] 2) LB medium:

[0030] Liquid LB medium (LLB): 10g L -1 Peptone, 5g·L -1 Yeast extract, 10g·L -1 NaCl, pH 7.2.

[0031] Solid LB medium (SLB): 1000 mL LLB + 15 g agar, pH 7.2.

Embodiment 2

[0032] Embodiment 2, the acquisition of bacterial strain

[0033] 1) Screening: collect 1g of clover rhizosphere soil not polluted by PCBs, break it up with 10mL sterile water, take 1mL of the suspension in 100mL of LSM, and culture it at 28°C and 150rpm for 5-7d; take 1mL of the bacterial suspension in In the new 100mLSM, transfer 5 times;

[0034] 2) Isolation: Take 200 μL of the bacterial suspension from the last subculture and spread it in SSM, and culture it at 30°C for 4 days to obtain a single colony;

[0035] 3) Purification: Pick a single colony and streak it on the SLB medium for 3 times to obtain pure strains;

[0036] 4) Obtaining: the pure strain in 3) is made into a bacterial suspension, and spread on an SSM plate. Cultivate it statically at 28°C, select a strain that grows fast and well, and name it SYC01, and store it on the SLB slope.

Embodiment 3

[0037] Embodiment 3, the identification of bacterial strain

[0038] 1. Morphological and physiological and biochemical characteristics

[0039] The strain SYC01 was inoculated on the SSM plate and cultured statically for 4 days at 28°C. The morphological characteristics of the colonies and cells were observed, and physiological and biochemical experiments were carried out.

[0040] The observed and experimental results are as follows:

[0041]1) Colony characteristics: The diameter of the colony is 0.5-1.0mm, the colony is round, the surface is smooth and moist, the edges are neat, slightly raised, and yellow.

[0042] 2) Cell morphological characteristics: the cells are rod-shaped with blunt apex; Gram staining is negative; the cell size is 0.5-0.6 μm×1.0-2.6 μm.

[0043] 3) Physiological and biochemical characteristics:

[0044]

[0045] 2. PCR amplification and sequence determination of the 16SrDNA gene of strain SYC01

[0046] Preparation of SYC01 total genomic DNA...

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Abstract

The invention relates to a strain having a symbiotic relationship with clover and degrading polychlorinated biphenyls, and a screening method thereof. The above bacterium is Pseudomonas sp. named as SYC01, and is preserved in China General Microbiological Culture Collection Center on May 7, 2015 with the preservation number of CGMCC No.10784. The strain is from microbes having a symbiotic relationship with rhizosphere of clover, and can degrade polychlorinated biphenyls (PCBs). The application of the Pseudomonas sp. having a symbiotic relationship with clover in degradation of the PCBs has not been reported. The strain has a symbiotic relationship with plant and plays a positive role in elimination and reduction of the PCBs in soil, prevention of harms of the PCBs to environment and organisms. The invention also provides a method for separating and screening the SYC01 strain from soil at non-PCBs pollution points and rhizosphere of clover through adopting biphenyls as the only carbon source and energy.

Description

technical field [0001] The present invention particularly relates to a strain of Pseudomonas ( Pseudomonas sp. ) SYC01 and its screening method. Background technique [0002] Polychlorinatedbiphenyls (PCBs) are in Fe 3+ Under the catalysis of the catalyst, the artificially synthesized chlorine-containing organic compounds with biphenyl as the matrix have a total of 209 homologues due to the different substitution positions of chlorine. As a chemical product with stable physical and chemical properties, it has been widely used in many industrial production fields in Europe, Asia, North America and other places. The total global production has exceeded 1.5 million tons, and many of them have been exposed to the environment. [0003] PCBs in the natural environment exhibit multiple sided characteristics. First, due to the presence of chlorine atoms, PCBs are resistant to biodegradation, with a half-life of several years or even decades in soil alone. Secondly, with the atmo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20B09C1/10C12R1/38
Inventor 胡星杜丽婷余应新牛莉莉胡雪峰刘建勇
Owner SHANGHAI UNIV
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