CN103409346B - A kind of heavy metal resistance Pectinatus and application thereof - Google Patents
A kind of heavy metal resistance Pectinatus and application thereof Download PDFInfo
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- CN103409346B CN103409346B CN201310312329.7A CN201310312329A CN103409346B CN 103409346 B CN103409346 B CN 103409346B CN 201310312329 A CN201310312329 A CN 201310312329A CN 103409346 B CN103409346 B CN 103409346B
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- heavy metal
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Abstract
Do you the invention discloses the bacterial strain Pectobacterium that a kind of heavy metal is resistance? sp.NP22.Does is this bacterial strain at the preserving number at China Committee for Culture Collection of Microorganisms's common micro-organisms center: CGMCC? NO.7794; The 16S of this bacterial strain? is rDNA sequence as SEQ? ID? described in NO.1.This bacterial strain can tolerate Cd
2+, Mn
2+, Zn
2+, Fe
2+, Cu
2+etc. various heavy, especially to Cd
2+, Mn
2+, Zn
2+tolerable concentration is very high, reaches Cd respectively
2+2000mg/L, Mn
2+3200mg/L, Zn
2+3200mg/L.Can prepare heavy metal absorbent by after actication of culture, heavy metal cadmium maximal absorptive capacity reaches 113mg/g.This bacterium can be used for making heavy metal adsorption, in heavy metal reparation and recovery, have using value.
Description
Technical field
The invention belongs to biological chemical field, be specifically related to a kind of heavy metal resistance Pectinatus and application thereof.
Background technology
Along with the development of human society industrial economy and the increase of the biochemical articles for use of agricultural, the problem of heavy metal contamination is day by day sharp-pointed, one of great environmental problem becoming global concern.Heavy metal contamination can cause agricultural land soil Fertility Degeneration, and quality of agricultural product reduces, and accelerates ambient water quality deterioration, and by food chain, finally has influence on human health.Cadmium is the heavy metal that a kind of toxicity is very large, and its compound also mostly belongs to toxicant.Cadmium is had many uses, and pigment, alloy, plating, battery, metallurgy etc. all will use cadmium.Along with the development of modern industrial and agricultural production, the use of the discharge of " three wastes ", sewage irrigation and agricultural chemicals, weedicide and chemical fertilizer gets more and more, the content of Cadmium in Soil significantly increases, Cd pollution problem in soil-plant-environmental system is increasingly severe, thus makes the security of agricultural-food be subject to serious threat.Just because of more such reasons, international and the domestic research for heavy metal contamination and control thereof has become the emphasis and hot research field that environment, chemistry, life science etc. intersect, and related application Journal of Sex Research lays particular emphasis on the aspects such as source that analysis heavy metal pollutes, prophylactico-therapeutic measures and environment remediation.
Administer heavy metal contamination both at home and abroad at present, mainly adopt the physico-chemical process that ion-exchange, chemical precipitation, solvent extraction etc. are traditional.Owing to needing complicated facilities and equipment and destroying soil layer construction, the cost of investment of physical chemistry repairing method is higher, and the matrix character that they also can affect this life of soil even can spoiled soil diversity of organism, so should not use for the contaminated soil of big area.But bioremediation technology is pulled within the concern scope in people face gradually because of its exclusive advantage.Biological restoration refers to that all take biology as the technology that main body is curbed environmental pollution.It comprises the pollutent utilized in animal, plant and antimicrobial absorbent, conversion, degraded soil and water, the concentration of pollutent in environment is reduced, or be other pollution-free materials by contamination transform, and make pollutent stabilization, avoid it to spread in environment.In Study on Environmental Protection, microorganism remediation also becomes the interest place of Many researchers.Research for microorganism heavy metal resistance and enrichment utilizes microorganism remediation heavy metal contamination good approach.Because the research of this respect both can provide technical support for microorganism remediation heavy metal contamination, also can promote that people understand the mechanism of microorganism heavy metal resistance, thus carry out environment remediation and ecological protection better.The application and development utilizing Pectinatus to carry out heavy metal adsorption has no other people report, and this patent provides a strain heavy metal resistance Pectinatus and the application in heavy metal adsorption thereof.
Summary of the invention
The object of the present invention is to provide a kind of heavy metal resistance Pectinatus and application thereof.Can be used for heavy metal absorbent by after actication of culture, heavy metal cadmium maximal absorptive capacity reaches 113mg/L.This bacterium can be used for making heavy metal absorbent, in heavy metal reparation and recovery, have using value.
Technical scheme of the present invention is: a kind of heavy metal resistance Pectinatus, and described bacterial strain at the preserving number at China Committee for Culture Collection of Microorganisms's common micro-organisms center is: CGMCCNO.7794; Preservation date on June 21st, 2013, Classification And Nomenclature is Pectobacteriumsp., and bacterial strain 16SrDNA sequence is as described in SEQIDNO.1.
The preparation method of above-mentioned heavy metal resistance Pectinatus is:
(1) sample separation from soil, sample adds in 100mLLB liquid nutrient medium, in 250mL Erlenmeyer flask, cultivates 48h and coats on LB solid plate, obtain bacterial strain with 120r/min rotating speed in 28 DEG C; Described LB liquid nutrient medium component is: containing peptone 10g in every 1L distilled water, yeast powder 5g, NaCl10g; Described LB solid plate component is: containing peptone 10g, yeast powder 5g, NaCl10g agar powder 12g in every 1L distilled water;
(2) by colony inoculation in containing cadmium screening culture medium, in incubator, cultivate 2d, screening Cadmium resistance bacterium, substratum used is: containing 50mg/L, peptone 10g, yeast powder 5g in every 1L substratum, NaCl10g, agar powder 12g, supply volume to 1L with distilled water;
(3) cadmium concentration in substratum is improved constantly until Cd
2+concentration is 2000mg/L, obtains the bacterial strain of resistance to cadmium.
(4) to bacterial 16 S rDNA gene clone and sequential analysis; Upstream primer sequence for cloning bacteria 16SrDNA gene is: AGAGTTTGATCCTGGCTCAG, and downstream primer sequence is: GGTTACCTTGTTACGACTT.
The invention has the beneficial effects as follows: with containing Cd
2+concentration is the screening culture medium of 2000mg/L, screens and identify heavy metal resistance Pectinatus NP22 of the present invention from soil.Can be used for heavy metal absorbent by after actication of culture, heavy metal cadmium maximal absorptive capacity reaches 113mg/g.This bacterium can be used for making heavy metal absorbent, in heavy metal reparation and recovery, have using value.
Accompanying drawing explanation
Fig. 1 is the bacterium colony picture of Pectinatus NP22.
Fig. 2 is the agarose gel electrophoresis figure of bacterial 16 S rDNA.(left side is Marker:D2000, and a size is 2000bp, 1000bp, 750bp, 500bp, 250bp, 100bp from top to bottom; The right is 16SrDNA).
Embodiment
1, material
(1) bacterial strain
Bacterial strain Pectobacteriumsp.NP22 (CGMCCNO.7794), described bacterial strain Pectobacteriumsp.NP22 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preserving number: CGMCCNO.7794.
(2) plant and instrument
American AB I company PCR amplification instrument, U.S. BIO-RAD company GelDocXR+ gel imaging system, Germany SIGMA company Sigma3-16pk table-type high-speed refrigerated centrifuge, SANYO GS Electronics Co., Ltd. Ultralow Temperature Freezer, Changzhou Guohua Electric Appliance Co., Ltd.'s low speed centrifuge, Shanghai Bo Xun Industrial Co., Ltd. YXQ-LS-75 vertical pressure steam sterilization pan, Shanghai Yue Ping scientific instrument company limited FA2004 electronic balance, upper Nereid grand experimental installation company limited DNP-9162 type electro-heating standing-temperature cultivator, Beijing Liuyi Instrument Factory DYY-12 type electrophoresis apparatus, Shanghai Precision Scientific Apparatus Co., Ltd 7230G visible spectrophotometer.
2, a kind of screening of heavy metal resistance Pectinatus
(1) sample separation from soil, sample adds in the LB liquid nutrient medium of 100mL, in 250mL Erlenmeyer flask, cultivates 48h and coats on LB solid plate, obtain bacterial strain with 120r/min rotating speed in 28 DEG C; Described LB liquid nutrient medium component is: containing peptone 10g in every 1L distilled water, yeast powder 5g, NaCl10g; Described LB solid plate component is: containing peptone 10g, yeast powder 5g, NaCl10g agar powder 12g in every 1L distilled water;
(2) by colony inoculation in containing cadmium screening culture medium, in incubator, cultivate 2d, screening Cadmium resistance bacterium, substratum used is: containing 50mg/L, peptone 10g, yeast powder 5g in every 1L substratum, NaCl10g, agar powder 12g, supply volume to 1L with distilled water;
(3) improve constantly cadmium concentration in substratum until Cd2+ concentration is 2000mg/L, screening obtains the bacterial strain of resistance to cadmium.Fig. 1 is shown in by the colonial morphology photo of bacterial strain.
3, identification of strains analysis
(1) gramstaining
Bacterium is coated with on slide glass; First dye: drip Viola crystallina to Bacterial stain 1-2min, water-wash away dye liquor; Mordant dyeing: wash away residual water with iodine liquid, and use iodine staining 1min, water-wash away dye liquor; Decolouring: suck the residual water on slide with filter paper, tilts slide, under white background, adds the ethanol decolorization of 95% with dropper stream, until the ethanol flowed out is without purple, washes (bleaching time is generally 20-30s) immediately; Redye: redye about 2min with sarranine liquid, washing; Microscopy: after drying, observes with oily mirror.It is gram-positive microorganism that thalline is dyed to hepatic, and what be dyed to redness is Gram-negative bacteria.
(2) bacterial strain 16SrDNA sequence and classification
Get 1mL bacterium liquid with the centrifugal 1min of 10000 × g, precipitation thalline; Abandon supernatant, add 1mL0.9%NaCl solution by pellet resuspended, the centrifugal 3min of 10000 × g; Abandon supernatant, add 450 μ L water, resuspension, add 50 μ L20%SDS, piping and druming evenly gently, 75 DEG C of water-bath 5min (becoming clarification to bacterium liquid); Phenol by 500 μ L3:1: chloroform adds and is wherein used for extracted protein, the centrifugal 5mi of 10000 × g; By the centrifuge tube of new for supernatant liquor to 1.5mL, supply 500 μ L, add the phenol of 500 μ L3:1: chloroform is extracting again; Draw supernatant and supply 500 μ L to new 1.5mL centrifuge tube, add 500 μ L chloroforms, mix gently, the centrifugal 5min of 10000 × g; The 1.5mL centrifuge tube that Aspirate supernatant to is new, adds to 500 μ L, adds the 3MNaAc of 1/10th volumes, then add the Virahol of same volume, put upside down several times, centrifugal 5min under 10000 × g condition; By supernatant liquor sucking-off, add 1mL70% ethanol for washing and precipitating, under 10000 × g condition, centrifugal 5min, removes supernatant, drying at room temperature; Add 50 μ LTE buffered soln dissolution precipitations, get 2 μ L electrophoresis to detect DNA;
To bacterial 16 S rDNA gene clone and sequential analysis; Upstream primer sequence for cloning bacteria 16SrDNA gene is: AGAGTTTGATCCTGGCTCAG, and downstream primer sequence is: GGTTACCTTGTTACGACTT; PCR reactions steps is: 1. 95 DEG C of denaturation 5min, 2. 95 DEG C of sex change 30s, 3. 54 DEG C of annealing 60s, and 4. 72 DEG C extend 90s, repeating step 2-4,30 circulations, and 5. 72 DEG C extend 10min." Shanghai Sangon Biological Engineering Technology And Service Co., Ltd " is sent by 16SrDNA to check order.Use blast program to carry out sequence alignment in GenBank, analyze strain classification status, and the Evolvement of Analysis and Screening bacterial strain and Sequences similar bacterial strain.Adopt pcr amplification to obtain 16SrDNA electrophoresis result and see Fig. 2.
4, bacterial strain heavy metal maximum tolerated concentration
Different concns Mn is added respectively in LB substratum
2+, Zn
2+, Fe
2+, Cu
2+substratum, the detection substratum of preparation respective concentration, analyzes the resistance of bacterial strain heavy metal.
Table 1NP22 heavy metal resistance cartogram
5, the preparation of bacterial strain heavy metal absorbent and adsorption effect
To can be used for after actication of culture preparing heavy metal absorbent, preparation method be: be inoculated in LB liquid medium by described bacterial strain from-80 DEG C of refrigerators, cultivates 16h with 120r/m rotating speed in 28 DEG C; The bacterial classification grown is transferred in new LB liquid medium with the amount that percent by volume is 5%, cultivates 16h with 120r/min rotating speed in 28 DEG C, obtain the bacterium liquid of logarithmic phase.With the centrifugal 10min of 6000r/min, supernatant discarded, collects thalline; The deionized water wash thalline of use sterilizing 3 times, is dried to constant weight by thalline, is prepared into heavy metal absorbent in 75 DEG C of baking ovens.
Adsorption effect: described heavy metal absorbent being added concentration is 2000mg/LCd
2+, pH7 solution in, at 28 DEG C of absorption 60min, collected by centrifugation supernatant, detects in solution and remains Cd
2+, detected result shows, and described heavy metal absorbent is to Cd
2+maximal absorptive capacity reaches 113mg/g.This bacterium can be used for making heavy metal absorbent, in heavy metal reparation and recovery, have using value.
Claims (2)
- Pectinatus that 1. a heavy metal is resistance ( pectobacteriumsp) NP22, it is characterized in that: described bacterial strain at the preserving number at China Committee for Culture Collection of Microorganisms's common micro-organisms center is: CGMCCNO.7794; The 16SrDNA sequence of bacterial strain is as described in SEQIDNO.1.
- Pectinatus that 2. a heavy metal is resistance ( pectobacteriumsp) application of NP22, it is characterized in that: be used for heavy metal absorbent by after actication of culture, heavy metal cadmium maximal absorptive capacity reaches 113mg/g.
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| CN106834159B (en) * | 2016-11-24 | 2019-08-16 | 华南农业大学 | One plant of HS233 bacterial strain and its application in resistance to cadmium and/or the effective cadmium content of reduction |
| CN107815428B (en) * | 2017-11-14 | 2020-06-09 | 四川农业大学 | Cadmium-removing rhizobium KG2, microbial inoculum containing rhizobium and application of microbial inoculum |
| CN109652346B (en) * | 2019-02-19 | 2020-11-10 | 北京本农科技发展有限公司 | Application of zinc adsorption strain and culture method of zinc adsorption strain |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102533616A (en) * | 2012-02-23 | 2012-07-04 | 东华大学 | Ralstonia pickettii highly resistant to cadmium and extraction method and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102533616A (en) * | 2012-02-23 | 2012-07-04 | 东华大学 | Ralstonia pickettii highly resistant to cadmium and extraction method and application thereof |
Non-Patent Citations (5)
| Title |
|---|
| Chromate reduction and 16S rRNA identification of bacteria isolated from a Cr(VI)-contaminated site;Pattanapipitpaisal P等;《App Microbiol Biotechnol》;20011231 * |
| Pectobacterium sp. NP25 16S ribosomal RNA gene, partial sequence;等;《Genebank》;20100118 * |
| 一株高抗镉细菌KCd1的分离鉴定及其抗性基因初步研究;曾艳等;《四川农业大学学报》;20100331;第28卷(第1期) * |
| 抗重金属微生物的筛选及其抗镉机理和镉吸附特性研究;曾晓希;《中国博士学位论文全文数据库》;20101231 * |
| 牛蒡根际可培养细菌多样性和镉耐性初步分析;侯进慧等;《生物技术通报》;20121231(第8期);158-162 * |
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