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Bacillus megaterium and application thereof in treating heavy metal-glyphosate complex pollution

A technology of Bacillus megaterium and heavy metals, applied in the field of microorganisms, can solve the problems of ineffective treatment of heavy metal-glyphosate complex pollution and achieve good degradation effect

Active Publication Date: 2017-08-18
QINGDAO AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Aiming at the above-mentioned problem that the prior art cannot effectively deal with heavy metal-glyphosate complex pollution, the present invention aims to provide a heavy metal-resistant Bacillus megaterium that efficiently degrades glyphosate wastewater, and utilizes this Bacillus megaterium to degrade the water body A method for glyphosate pollutants to solve the problem of microbial degradation of glyphosate in heavy metal polluted environments

Method used

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  • Bacillus megaterium and application thereof in treating heavy metal-glyphosate complex pollution
  • Bacillus megaterium and application thereof in treating heavy metal-glyphosate complex pollution
  • Bacillus megaterium and application thereof in treating heavy metal-glyphosate complex pollution

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Effect test

Embodiment 1

[0038] Embodiment 1, the screening of Bacillus megaterium PP84

[0039] (1) Isolation and purification of strains

[0040]1. Enrichment of bacterial strains: Collect 2.5g of farmland soil samples that have been applied with glyphosate for a long time, place them in a shake flask containing 60mL enrichment medium, add 30mL of 100mM glyphosate and 10mL of 5mM CuCl2 solution prepared by sterile water , placed in a shaker, 30 ° C, 180 rpm, constant temperature shaking culture for 4 days.

[0041] 2. Acclimatization of bacterial strains: under aseptic conditions, add the bacterial solution to the inorganic salt basic liquid medium with glyphosate as the only carbon source at an inoculum size of 10%. Increased to 500mM, from 5mM, 10mM to 15mM. The total volume of the culture solution is 100mL, placed in a shaker, 30°C, 180rpm, constant temperature shaking culture, 4d is one acclimatization cycle, and three cycles of acclimatization are carried out.

[0042] 3. Purification of str...

Embodiment 2

[0049] Example 2 Tolerance and degradability of bacterial strain PP84 to glyphosate

[0050] Under sterile conditions, the purified strain PP84 was pre-cultured in a basal salt medium containing 200mM glyphosate for 24 hours (OD ≈ 1.0), and the culture solution was inoculated into the basal salt medium with an inoculum volume fraction of 5%. , adding different concentrations (100, 250 and 500 mM) of glyphosate as a carbon source, respectively, 30 ° C, 180 rpm, shaking culture for 4 days. Samples were taken every 6 hours to determine the growth of bacteria and the degradation rate of glyphosate. Use an ultraviolet spectrophotometer to measure the OD value of the bacteria at a wavelength of 600 nm, and draw a growth curve. The concentration of glyphosate in the supernatant was determined by nitrosylation ultraviolet spectrophotometry.

[0051] Calculation method of degradation rate: degradation rate (%)=(1-C1 / C0)×100. C1: Glyphosate residual concentration in degrading bacteri...

Embodiment 3

[0054] Degradation of glyphosate by bacterial strain PP84 under different pHs in embodiment 3

[0055] Under sterile conditions, the purified strain PP84 was pre-cultured in a basal salt medium containing 200mM glyphosate for 24h (OD ≈ 1.0), and the culture solution was inoculated to a pH of 5.0 according to an inoculum volume fraction of 5%. 6.0, 7.0, 8.0, 9.0 In the inorganic salt medium with 250mM glyphosate as the sole carbon source, 30°C, 180rpm, constant temperature shaking culture for 56h, the degradation rate of glyphosate was measured. The determination of the glyphosate degradation rate is the same as that of the implementation case 2.

[0056] Depend on Image 6 It can be seen that the strain PP84 grows in a wide pH range in the culture medium with glyphosate as the sole carbon source, grows well in the range between pH 4.0 and pH 9.0, and has good tolerance to glyphosate. Degradation. The optimum pH range for strain PP84 to degrade glyphosate was pH5.0-8.0, and ...

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Abstract

The invention provides a Bacillus megaterium Bacillus megaterium PP84. Deposition unit: China microorganism preservation management committee general microbiological center (CGMCC); address: Institute of Microbiology, No. 3, academy, No. 1, Beichen West Road, Chaoyang district, Beijing, China; deposition date: July 20, 2016; deposition number: CGMCC No. 12798. The Bacillus megaterium Bacillus megaterium PP84 has the advantages of having the capacity of using glyphosate as carbon and nitrogen sources; having the resistance capacity on heavy metals; being capable of decomposing and assimilating glyphosate, and adsorbing the heavy metals simultaneously; achieving the purpose of degrading the glyphosate in the environment.

Description

technical field [0001] The invention belongs to the field of microorganisms, and in particular relates to a Bacillus megaterium strain with resistance to heavy metals and its degradation of glyphosate in a heavy metal polluted environment. Background technique [0002] Glyphosate (N-phosphonic acid methyl-glycine) is a systemic broad-spectrum herbicide, which is widely used in agriculture because of its high efficiency and remarkable effect. However, studies in recent years have shown that long-term application of glyphosate will cause inestimable impacts on non-target organisms and the environment, especially on aquatic organisms and soil microorganisms. In addition, with the development of modern industry and agriculture, more and more heavy metals have entered the environment, and the frequent use of sewage irrigation, pesticides and fertilizers containing heavy metals has further deteriorated the environment. These production methods may lead to heavy metal-glyphosate c...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34C12R1/11C02F101/38C02F101/20C02F101/22
CPCC02F3/34C02F2101/20C02F2101/22C02F2101/306C02F2101/38C12N1/205C12R2001/11
Inventor 宋宁宁王芳丽王凯荣刘君李绍静贾存珍
Owner QINGDAO AGRI UNIV
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