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Identification identification and application of heavy metal tolerant penicillium in coal gangue

A heavy metal and Penicillium technology, applied in fungi, microorganism-based methods, and the restoration of polluted soil, etc., can solve the problem of microorganisms unable to reproduce, and achieve the effect of strong resistance to heavy metal cadmium and strong tolerance.

Inactive Publication Date: 2014-07-16
CHINA UNIV OF MINING & TECH (BEIJING)
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although a variety of heavy metal-resistant fungi have been isolated, most microorganisms cannot reproduce due to the strong acid wastewater containing high concentrations of heavy metals produced by weathering and leaching of high-sulfur coal gangue. Bacteria have not been reported

Method used

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  • Identification identification and application of heavy metal tolerant penicillium in coal gangue
  • Identification identification and application of heavy metal tolerant penicillium in coal gangue
  • Identification identification and application of heavy metal tolerant penicillium in coal gangue

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Embodiment 1

[0014] Embodiment 1: the separation of Penicillium of the present invention.

[0015] 12 samples were randomly selected from the Xiejiaji coal mining area in Huainan to form a representative sample for enrichment and screening. Add 5g gangue sample into 45mL Martin's liquid medium, enrich and cultivate for 48 hours at 28°C and 180r / min, then take 2mL of enriched bacteria liquid and inoculate it with 100mg / L Cd 2+ cultured in the fresh medium of 200mg / L Cd 2+ cultured in liquid medium, and so on, increasing Cd step by step 2+ Concentration, at 300-5000mg / L Cd 2+ cultured in culture medium. Take 0.2mL1000mg / L Cd 2+ The bacterial solution was coated in 5000mg / L Cd 2+ Inverted on the solid medium of 28 ° C incubator for 120 h to isolate Cd-resistant strains. According to the preliminary observation of the colony morphology, the BJKD4 monoclonal strain was mold, and it was inoculated to 500~3500mg / L Cd 2+ The bacteria were domesticated and cultured in Chapeauer's medium, and...

Embodiment 2

[0019] Embodiment 2: 18S rDNA sequence determination of Penicillium BJKD4 bacterial strain of the present invention

[0020] 18S rDNA sequence analysis of the strain: the genome was extracted by CTAB (cetyltrimethylammonium bromide) method. DNA was amplified by polymerase chain reaction (PCR) using amplification primers: SSU817TTA GCA TGG AAT AAT RRAATA GGA and SSU1536AAT GCAATG CYC TAT CCC CA. The 50μL reaction system contains: 5μL of 10*PCR buffer, 0.25L of 10mmol / L Dntp, 0.5μL of 45pmol / μL primer, 0.5μL of 2.5U / μL Taq enzyme, 1μL of DNA template and 42.25μL of sterilized deionized water. PCR reaction conditions: denaturation at 94°C for 1 min, annealing at 50°C for 30 s, extension at 72°C for 90 s, 40 cycles, and final extension at 72°C for 5 min. 2 μL of the PCR reaction product was electrophoresed on 1% agarose gel. The DNA sequence was determined by Beijing Huada Gene Company, using BLAST ( http: / / www.ncbi.nlm.nih.gov / BLAST ) program for DNA sequence alignment.

[0...

Embodiment 3

[0022] Embodiment 3: the determination of the ability of penicillium of the present invention to resist heavy metals

[0023] 1. Tolerance of BJKD4 bacterial strains to heavy metal cadmium: inoculate the bacterial strains activated for 2 days into 0, 1, 3, 9, 18 or 36mmol / LCd 2+ In the solid Chapeau medium, cultured at 28°C for 16 days, and measured the colony diameter on the 3rd day.

[0024] The test results showed that the BJKD4 strain contained 0-36mmol / L Cd 2+ Grow in the Chapei solid medium for 16 days, the colony diameter increases with the prolongation of the culture days, and with the Cd 2+ decrease with increasing concentration ( Figure 4 ). In no Cd 2+ In the medium (control), the lag phase of bacterial colony growth is 0-4d, the logarithmic growth phase is 4-15d, and enters the stable phase at 16d; while adding heavy metal Cd 2+ (1-36mmol / L), the lag phase extends to the 6th day, and then enters the logarithmic phase, and reaches a stable phase at 14-15d. Co...

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Abstract

The invention discloses a heavy metal cadmium tolerant penicillium BJKD4 (Penicillium sp.) screened from coal gangue in a mining area. The strain is preserved in China General Microbiological Culture Collection Center (CGMCC) with a preservation number of CGMCC NO.6620. The Penicillium BJKD4 strain provided by the invention has strong tolerance to cadmium salts and compound pollution of heavy metals, and plays an important role in reclamation of coal gangue mountains and ecological restoration of soil heavy metal pollution.

Description

Technical field: [0001] The invention relates to the technical field of bioremediation for controlling heavy metal pollution, in particular to the separation and application of a heavy metal cadmium-resistant penicillium. Background technique: [0002] The weathering and leaching of high-sulfur coal gangue can produce a strong acidic leaching solution, resulting in the leaching of heavy metals. The acidic wastewater containing heavy metals flows along the runoff of the gangue pile and migrates to the surrounding soil and water bodies, resulting in the deterioration of the gangue pile and the surrounding soil. And water bodies are severely acidified and polluted by heavy metals, such as cadmium, copper, lead, zinc, etc. Some metals are indispensable elements for the life activities of organisms, but excessive heavy metals have potential toxic effects. Berny P et al. reported a series of serious harms caused by heavy metals to humans, animals, plants and microorganisms. The p...

Claims

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Application Information

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IPC IPC(8): C12N1/14A62D3/02B09C1/10C12R1/80A62D101/43
Inventor 张玉秀柴团耀李霞
Owner CHINA UNIV OF MINING & TECH (BEIJING)
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