Aerobic arsenic methylation bacterium SM-1 of cytophagaceae and application of arsenic methylation bacterium

A fibrous and methylation technology, applied to aerobic arsenic methylating bacteria SM-1 and its application fields, can solve the problems of difficult arsenic pollution control, no soil culture experiments, and no ability to volatilize arsenic. , to achieve the effect of reducing the harm

Inactive Publication Date: 2016-04-13
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these two strains of bacteria have been confirmed to have the ability to methylate arsenic, the researchers also identified a trivalent arsenic methyltransferase gene (arsM) from their respective bodies, but neither of these two strains has the ability to volatilize arsenic , neither of them can produce volatile arsine to remove arsenic from the soil, and neither of these two strains has carried out corresponding soil culture experiments to prove their effects in the soil ecological environment system, so it is difficult for them to be really applied to rice fields Treatment of Soil Arsenic Pollution

Method used

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  • Aerobic arsenic methylation bacterium SM-1 of cytophagaceae and application of arsenic methylation bacterium
  • Aerobic arsenic methylation bacterium SM-1 of cytophagaceae and application of arsenic methylation bacterium
  • Aerobic arsenic methylation bacterium SM-1 of cytophagaceae and application of arsenic methylation bacterium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Enrichment of soil samples and isolation, purification and preservation of arsenic methylating bacteria SM-1

[0026] Take 2 g of paddy soil from Shimen arsenic-contaminated areas and add it to 100 mL of enrichment medium. The composition of the enrichment medium is (g / L): ST10 -1 (0.5g peptone, 0.05g yeast extract), 0.5% glucose, 10 μMAs (III). The medium was placed in a shaker at 200 rpm and 30 °C for one week, and 5 mL of the enrichment solution was taken out after one week and transferred to a new 100 mL enriched culture set. The culture was continued under the same culture conditions. The above transfer process was repeated twice. Finally, 2 mL of the enriched solution was taken out for speciation analysis of arsenic. Before the samples were used for detection, they were centrifuged at 12,000 rpm for 2 min, and then passed through a 0.22 μM filter. High performance liquid chromatography-inductively coupled plasma mass spectrometer (HPLC-ICP-MS) was used...

Embodiment 2

[0027] Example 2: Identification of bacterial species

[0028] (1) Determination of physiological and biochemical properties:

[0029] Table 1. Physiological and biochemical properties of strain SM-1

[0030]

[0031] Note: + means available; ‐ means not available

[0032] (2) Molecular biological identification

[0033]Use the CTAB method to extract the total bacterial DNA, use the DNA as a template, and use the universal primers F27 and R1492 PCR to amplify the 16SrRNA gene of the strain, recover and purify the PCR product of about 1500bp, connect it with the carrier PMD18T and transform it into Escherichia coli In DH5α, positive clones were selected and sent to the sequencing company for sequencing, and the returned sequence was input into NCBI for Blast comparison with other 16SrRNA gene sequences. The sequence has been submitted to NCBIGenBank with accession number KT989310. Combined with the results of the above physiological and biochemical characteristics and 16...

Embodiment 3

[0034] Example 3: Detection of arsenic methylation and volatilization ability of bacterial strains under laboratory conditions

[0035] R 2 The components of medium A are (g / L): yeast powder 0.5g, tryptone 0.5g, casamino acid 0.5g, glucose 0.5g, soluble starch 0.5g, dipotassium hydrogen phosphate 0.3g, magnesium sulfate heptahydrate 0.05 g, sodium pyruvate 0.3g. Inoculate the strain SM-1 in 100mL of the above-mentioned liquid medium, and cultivate it on a shaker at 37°C and 200rpm for one day and night to make the OD of the bacteria 600 When the value reaches 1.0, 1 mL of the seed solution is taken out from the seed medium and added to a new 100 mL of R with 10 μM As(III). 2 In liquid culture medium A, the treatments were repeated three times, and the transfer solution was continued to be cultured under the same culture conditions as above, and a solution sample was taken every 4 hours. After the taken solution was centrifuged at 12000rpm for 2min, the supernatant was filter...

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Abstract

The invention discloses an aerobic arsenic methylation bacterium SM-1 of cytophagaceae and application of the arsenic methylation bacterium. A gram-negative bacterium, which is quite strong in arsenic methylation and volatilization capacities, is obtained through isolation and identification; and the strain belongs to a new genus and a new species of the cytophagaceae, which is named as arsenicibacter palustris SM-1 and preserved in China Center for Type Culture Collection on October 15, 2014 with preservation number of CCTCC M2014457. Compared with the prior art, the bacterium disclosed by the invention has the following beneficial effects: the provided bacterium strain is quite strong in arsenic methylation and volatilization capacities; and the strain is applicable to arsenic pollution remediation of paddy soil and is capable of finally relieving the damage from arsenic pollution in a paddy field.

Description

technical field [0001] The invention relates to the technical field of environmental microorganisms, in particular to an aerobic arsenic methylated bacterium SM-1 belonging to the family Cellophila and its application. Background technique [0002] Arsenic (As) is a toxic and harmful metalloid element, and also a terrible carcinogen. More than one million people around the world are poisoned by this harmful element. In recent decades, human production activities such as mining, chemical smelting, and burning of fossil fuels have exacerbated the spread and diffusion of arsenic in water, atmosphere, and soil environments. In Southeast Asia, a region where rice is the main diet, the situation of arsenic pollution in rice fields is increasing day by day. The high concentration of arsenic in paddy soil will not only affect the growth and yield of rice, but also enter the human body through the accumulation of rice, causing health hazards to people who eat it for a long time. G...

Claims

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Application Information

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IPC IPC(8): C12N1/20B09C1/10C12R1/01
CPCB09C1/10C12N1/205C12R2001/01
Inventor 赵方杰黄科
Owner NANJING AGRICULTURAL UNIVERSITY
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