CN106086154A - Carbon nanomaterial regulation and control lawn composting substrate Fungal identification is used to analyze method - Google Patents
Carbon nanomaterial regulation and control lawn composting substrate Fungal identification is used to analyze method Download PDFInfo
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Abstract
The invention discloses a kind of carbon nanomaterial regulation and control lawn composting substrate Fungal identification and analyze method, with consumer garbage compost and soil mixture for lawn matrix material, to mix four kinds of different carbon nanomaterials wherein as process group, cultivated by lawn plant, substrate can be cultivated after research interpolation different carbon nanomaterial a period of time the change of fungus monoid, and analyzed the classification position that can cultivate fungus in substrate by molecule sequencing technologies.By carbon nanomaterial being added to the understanding of the change of fungus monoid after turf establishment system, provide technical support for carbon nanomaterial fungus monoid regulation and control application in garbage compost lawn matrix.
Description
Technical field
The invention belongs to environmental protection technical field, relate to a kind of employing carbon nanomaterial regulation and control lawn composting substrate fungus
Identification and analysis method.
Background technology
Carbon nanomaterial is an at least material less than 100 nm in three dimensional structure.Carbon nanomaterial has size
Little, specific surface area is big, and surface can be high, and the feature that surface atom ratio is big shows at aspects such as mechanics, optics, calorifics, electricity
Excellent properties.There is a lot of micro-interface in nano-particle, various interfacial reactions can be strengthened by it, dirty at heavy metal-polluted soil
Dye and sewage purification aspect play obvious action.
Carbon is the highest at nature content, the most extremely important for Organic substance and life entity.1985, scientist sent out
Show by 60 former molecular C of C60.CNT and Graphene etc. are found the most in succession subsequently.2004, Geim et al.
Successfully prepare single-layer graphene.Found have along with Graphene successively in the special nature in optical, electrical, thermal and magnetic afterwards
The research closing material with carbon element enters again a brand-new field.
Along with science and technology, being continuously increased of the fast development of modern industry and population, in the world a lot of countries all in
Reveal the trend that water body, soil and atmospheric pollution are the most serious.Report is pointed out, the river of China 50% and the lake of more than 80%
Pool has had been subjected to pollute, and many lakes of III class water standard China of surface water have not reached.The arable land in the whole nation
In have 80,000,000 hm2More than receive pollution in various degree, and distinct issues be exactly organic pollution and
The pollution of heavy metal, this situation the most seriously constrains the enforcement of Sustainable Development Strategy of China, has even threatened compatriots
Health.
Although traditional river system repair technology such as cuts dirt, Sediment Dredging etc. temporarily can play some effects, but all controls
Mark does not effects a permanent cure, and biological restoration limiting factor has a lot, such as ambient temperature and Acidity of Aikalinity etc., and this allows for biological restoration visitor
Significant limitation is there is in sight.As traditional contaminated soil remediation method, soil moved in improve the original method, leaching method waste time and energy, and
Well effect is not had for large-area reparation.Compared with traditional environment restoring method, nano material is used for weight
Metallic prosthetic, the absorbability superpower because of it and the specific surface area of super large, the shortcoming that the most there is not traditional method, also show
High remediation efficiency.Additionally atmospheric pollution aspect situation is also SO that is the severeest, that exceed standard in air2, CO and NOxIn the moment
Threaten our healthy.Nano material has splendid catalytic efficiency, even can be catalyzed the most non-incident reaction
New method and thinking is provided for solving generation atmosphere polluting problem.Therefore, utilize nano material solve water body and soil with
And the pollution problem of air is increasingly worth people to pay close attention to.And the research of carbon nanomaterial is the most promising neck
Territory.
Research in recent years finds that CNT can effective heavy metal ion in adsorbed water body.The experiment table of Li etc.
Bright CNT can lead ion in adsorbed water body, and have the biggest dependency with the pH value of solution.Along with solution ph
Increase, the absorbability of lead ion is increased by CNT the most therewith.Langmuir and Freundlich model can solve
Release this adsorption process.Subsequently, Li etc. are investigated the CNT of the different mode of oxidizing process adsorption effect to cadmium ion
Relatively, finding after oxidation processes, the specific surface area of CNT dramatically increases, and the quantity of surface functional group also has substantially increasing
Many.Specifically, CNT is through KMnO4Adsorption effect after oxidation is apparently higher than H2O2And HNO3Oxidation effectiveness.Separately
Outward, next Li et al. has carried out again the experiment of carbon nanotube adsorption copper ion, have studied the CNT of different shape to lead
The absorption of ion is compared, the CNT Adsorption Kinetics to lead ion and the situation of desorption, and discusses carbon
Nanotube is to lead ion, copper ion and cadmium ion competitive Adsorption situation in the same solution.Research finds, CNT pair
It is Pb to the absorption order of three of the above different ions in solution2+＞ Cu2+＞ Cd2+, wherein to lead ion (Pb2+), copper ion
(Cu2+) absorption can with Langmuir equation explain.
Li Yanhui etc. utilize HNO3The CNT of oxidation processes has been carried out Pb2+Adsorption experiment.Research finds HNO3
Carbon nano tube surface after oxidation processes is long-pending to be significantly increased, and additionally the functional group such as-OH ,-CO ,-COOH has been introduced in carbon and receives
The surface of mitron.This can be remarkably reinforced carbon nanotube adsorption Pb2+Active area and active force, thus improve CNT
To Pb2+Adsorbance.Result shows, adsorbance raises along with the rising of temperature, and this explanation CNT is to Pb2+Absorption
It it is an exothermic process.Regeneration test shows, adsorption effect dramatically increases with the increase of pH value.When pH as little as 2, carbon nanometer
The Pb of absorption on pipe2+Desorption rate is to 85%, thus provides theory depend on for realizing recycling of carbon nanotube adsorption material
According to.
In addition, Graphene and graphene oxide for Organic Pollution and are revealed the environmental pollution caused by oil
Also there is the strongest repair.After Graphene modification to the adsorption effect of pollutant in environment more preferably, and adsorbance increase,
Performance is more stable.Chen et al.]The three-dimensional porous graphene oxide film prepared shows huge in terms of selecting absorption
Big potentiality.The weight of its absorption machine oil can reach 37 times of own wt, and the weight of absorbing organic solvent is then more than self
26 times of weight, its adsorption effect compares flake graphite alkene and foamy graphite alkene is much higher.In addition, this porous oxidation
The character of graphene film is sufficiently stable, recycles after the adsorbent on available hexane removal surface.Higher absorption
Amount and longer recycling number of times make this three-dimensional porous graphene oxide film organics removal and cleaning oil pollution
Aspect shows great application prospect.
Above-mentioned analysis shows, the carbon nanomaterial such as Graphene and CNT environmental area application mainly soil,
Water body and atmospheric pollution reparation aspect, and apply as regulator, especially carbon nanomaterial is at compost lawn base
As microorganism adjusting control agent in matter, research there is no document report at present.
Along with the junk produced in China's economic growth and the quickening of urbanization speed, urban life and production process
Present the trend dramatically increased.Meanwhile, the supporting process of house refuse is the most complete, and a large amount of rubbish are exposed to city
In, environment is caused the biggest impact.Composting is urban garbage disposal and resource compares mode easily,
This can not only digest the house refuse in city, alleviates the urban environment pressure that house refuse is brought, and also produces simultaneously
The supply agricultural productions of a large amount of compost fertilizer.
Containing nutrient necessary to a large amount of plant growings in house refuse, therefore consumer garbage compostization is processed also
Compost is used for the effective way that agricultural production is its recycling.
Research finds to add appropriate garbage compost to be effectively increased crop yield in the middle of soil.Because of soil physical chemistry
Matter and agrotype are different, and its amplification there is also significant difference.Zhou Dezhi etc.]After have studied interpolation garbage compost, yellowish soil,
The wheat yield change of plantation in moisture soil and red soil.Result shows, after adding appropriate garbage compost, crop all has volume increase effect
Really, wherein red soil effect is best (crop yield 46%).He Liyuan etc.[43]Research finds, the yield of Plantula Brassicae chinensis is along with rubbish heap
The raising of fertile amount of application and increase, and difference reaches significant level compared with the control.Additionally, the using not only to making produce of compost
Amount has an impact, and also increase significantly effect to the quality of crop.The research of square pavilion et al. finds, adds garbage compost to palm fibre
In red soil, in the seed of soil crop Brassica campestris L, the content of protein significantly raises, and protein contained in soybean kernel is obvious
Increase.
But, people also began to recognize and did not contain only nutrient substance in garbage compost in the last few years, simultaneously possibly together with one
Quantitative heavy metal, if chronic administration can increase the content of heavy metal in soil.Therefore, compost is used for agricultural production may
A series of food-safety problem can be brought.Visible, avoiding food chain, to realize the recycling of garbage compost significant.No
Only such, for preventing compost utilization from causing heavy metal pollution of soil, even bring threat to ecological environment, must while application
The problem that heavy metal is repaired must be considered.
Traditional sod production is typically to utilize high-quality soil.The physical property of soil can be had a huge impact by this,
Fertility is caused to decline.Additionally during sod production, the topsoil soils of 2 more than cm can be pulled away, and fertile topsoil soils enters city
Ecosystem, and after sod production several times, agricultural land soil will become more and more barrenr, so that finally cannot enter again
Row agricultural production, this is waste great to land resources.If turf establishment is using consumer garbage compost as primary structure,
Municipal refuse can not only be digested, additionally it is possible to alleviate turf production and bring the destruction of topsoil soils.
Nanotechnology development in recent years is very rapid, all shows wide application at aspects such as material, information, environment
Prospect.Although there being some to study theoretical research, show that nano material can grow the shadow produced in various degree to microorganism
Ring, but apply its regulating and controlling effect, also there is no correlation technique report.
Fungus belongs to eukaryotic microorganisms, and widely, the multiformity of fungus is for biosphere for the distribution in nature
Stable state and balance play an important role, and fungus also provides substantial amounts of living resources for the mankind simultaneously.In terms of Biomass,
Fungus in soil account for part and parcel.Wherein having kind more than 200 is conditioned pathogen.Fungus can be decomposed in the middle of soil
Organic form humus, provide nutrient for plant, the growth of supply soil animal, and fungus be also Indices of Ecological
Important criterion.Farmland, meadow, forest land etc. are local, and we can easily find the figure of fungus, but due to its institute
The environment at place is different, and under different Environmental Factors, in soil, different rule all can occur in the bacterium colony composition of fungus, distribution
Rule.Research finds that fungus plays function miscellaneous in the environment, such as explains cellulose, chelated metal ions etc.,
These functions also make to be in fungal ecology environment the status of tool foot weight.
Summary of the invention
Carbon nanomaterial just has the heavy metal in the biggest range of application, adsorbed water body and soil because of its special structure
It it is the important application of one of which.This technology, from substrate microorganism angle, with consumer garbage compost and soil mixture is
Lawn matrix material, to mix four kinds of different carbon nanomaterials wherein as process group, is cultivated by lawn plant, and research adds
Substrate can be cultivated after adding different carbon nanomaterial a period of time the change of fungus monoid, and analyze base by molecule sequencing technologies
Matter can be cultivated the classification position of fungus.By carbon nanomaterial being added to the change of fungus monoid after turf establishment system
Understanding, provides technical support for carbon nanomaterial fungus monoid regulation and control application in garbage compost lawn matrix.
The invention discloses following technology contents for achieving the above object:
A kind of method using carbon nanomaterial regulation and control lawn composting substrate Fungal identification to analyze, it is characterised in that by following step
Suddenly carry out:
(1) experiment material
Experiment soil takes from Tianjin Normal University's campus 0-20 cm topsoil, and fundamental property is: water content 19.4%, pH
7.27, electrical conductivity 2250 S/cm, phosphorus organic, complete, full nitrogen are respectively 52.3,3.75,2.15 g/kg.
Experiment consumer garbage compost is from Tianjin little Dian destructor plant.PH is 7.49, electrical conductivity 2300 S/cm,
Phosphorus organic, complete, full nitrogen are respectively 132,6.81,25.1 g/kg.Heavy metal Cr, Cu, Pb, Zn, Cd content is respectively 703,
341、217、677、5.01 mg/kg。
Being purchased from Qiu Shi carbon black plant, Tianjin for examination nano carbon black (CB), particle diameter is 20 ~ 70 nm, and specific surface area is 1200 m2/
G, pH value is 7.0, utilizes KMnO before experiment4To its modification.
Graphene (G) microplate is purchased from Nanjing Ji Cang nanosecond science and technology company limited, and structure is black foil shape, microplate size
0.5-20 um, thickness 5-25 nm, specific surface area 40-60 m2/ g, density about 2.25 g/cm3, electrical conductivity 8000-10000 S/
M, phosphorus content > 99.5%.
Graphene oxide (GO) is purchased from Suzhou Heng Qiu nanometer company, for black or brownish yellow powder, average thickness 3.4-7
Nm, lamella diameter 10-50 μm, specific surface area 100-300 m2/ g, purity > 90%.
CNT (CNT) is purchased from Beijing and wins space high-tech new material technology company limited, diameter 20-40 nm, length
10-30 um ,-COOH content 1.43%, purity>90wt%, ashes<8wt%, specific surface area>110 m2/ g, conduction: > 102 s/
cm。
(2) experimental technique:
Measure soil moisture content after fetching earth immediately, be calculated the ratio of soil dry weight and weight in wet base.Proportionally calculate to obtain dry weight
Wet soil weight corresponding to 1500 g.Claim the wet soil of corresponding weight mixes with 30 g garbage composts, stir, loading internal diameter
In the plastic flowerpot of high 20 cm of 15 cm, fill 15 basins altogether.
Experiment sets 5 process, Graphene (G), graphene oxide (GO), CNT (CNT), modified Nano white carbon black altogether
(CB) weigh the ratio of 1% in dry ground to be added separately in mixed-matrix, with the process without carbon nanomaterial for comparison (CK),
Fully mix after adding carbon nanomaterial, every basin sowing tall fescue seed 5g.During cultivation, temperature is 19 ~ 27 DEG C, the wettest
Degree is 60 ~ 72%, and every day keeps the skin wet to soil, makes soil moisture be maintained at the 70% of water capacity.After cultivating 130d, will
In basin, 0-5cm deep soil takes out mixing, crosses 60 mesh sieves, and the preservation of-20 DEG C of refrigerator, for microorganism separation and analysis of accounts.
The present invention open employing carbon nanomaterial regulation and control lawn composting substrate Fungal identification analysis method that added further exists
Determine the application in terms of the composition of fungus colony.The composition of described fungus colony therein includes: Penicillium(Penicillium), trichoderma(Trichoderma), chaetomium globosum belong to(Arthrobacter), Leptosphaeria(Leptosphaeria), Fusarium(Fusarium), Eurotium (Aspergillus), the raw red shell Pseudomonas of light color(Bionectria ochroleuca)。
Nano-sized carbon Graphene (G) of the present invention, graphene oxide (GO), CNT (CNT), modified Nano white carbon black
(CB).Modified Nano white carbon black (CB), utilizes KMnO before experiment4Its modification is referred to: weigh nano-sized carbon 10 g in 250
In mL conical flask, add 100 mL0.03 mol L-1KMnO4Solution, after standing 10 min, is put on general-purpose electric heater boiling
Rise backflow 1 h.After cooling, repeatedly rinse with deionized water, make solution no longer muddy and pH is stable.It is transferred to beaker, 110 DEG C of bars
Dry to constant weight under part.
More detailed description of the present invention is as follows:
1 develops materials and methods
1.1 experiment material
Experiment soil takes from Tianjin Normal University's campus 0-20 cm topsoil, and fundamental property is: water content 19.4%, pH
7.27, electrical conductivity 2250 S/cm, phosphorus organic, complete, full nitrogen are respectively 52.3,3.75,2.15 g/kg.
Experiment consumer garbage compost is from Tianjin little Dian destructor plant.PH is 7.49, electrical conductivity 2300 S/cm,
Phosphorus organic, complete, full nitrogen are respectively 132,6.81,25.1 g/kg.Heavy metal Cr, Cu, Pb, Zn, Cd content is respectively 703,
341、217、677、5.01 mg/kg。
Being purchased from Qiu Shi carbon black plant, Tianjin for examination nano carbon black (CB), particle diameter is 20 ~ 70 nm, and specific surface area is 1200 m2/
G, pH value is 7.0, utilizes KMnO before experiment4To its modification.
Graphene (G) microplate is purchased from Nanjing Ji Cang nanosecond science and technology company limited, and structure is black foil shape, microplate size
0.5-20 um, thickness 5-25 nm, specific surface area 40-60 m2/ g, density about 2.25 g/cm3, electrical conductivity 8000-10000 S/
M, phosphorus content > 99.5%.
Graphene oxide (GO) is purchased from Suzhou Heng Qiu nanometer company, for black or brownish yellow powder, average thickness 3.4-7
Nm, lamella diameter 10-50 μm, specific surface area 100-300 m2/ g, purity > 90%.
CNT (CNT) is purchased from Beijing and wins space high-tech new material technology company limited, diameter 20-40 nm, length
10-30 um ,-COOH content 1.43%, purity>90wt%, ashes<8wt%, specific surface area>110 m2/ g, conduction: > 102 s/
cm。
Technology design
Measure soil moisture content after fetching earth immediately, be calculated the ratio of soil dry weight and weight in wet base.Proportionally calculate to obtain dry weight
Wet soil weight corresponding to 1500 g.Claim the wet soil of corresponding weight mixes with 30 g garbage composts, stir, loading internal diameter
In the plastic flowerpot of high 20 cm of 15 cm, fill 15 basins altogether.
Experiment sets 5 process, Graphene (G), graphene oxide (GO), CNT (CNT), modified Nano white carbon black altogether
(CB) weigh the ratio of 1% in dry ground to be added separately in mixed-matrix, with the process without carbon nanomaterial for comparison (CK),
Each process repeats for three times.Fully mix after adding carbon nanomaterial, every basin sowing tall fescue seed 5g.During cultivation, temperature is
19 ~ 27 DEG C, relative humidity is 60 ~ 72%, and every day keeps the skin wet to soil, makes soil moisture be maintained at water capacity
70%.After cultivating 130d, 0-5cm deep soil in basin is taken out mixing, cross 60 mesh sieves, the preservation of-20 DEG C of refrigerator, divide for microorganism
From with analysis of accounts.
Method of production
1.3.1 fungi culture medium compound method
Cultivating fungus rose bengal medium, concrete formula is peptone 5g, glucose 10g, potassium dihydrogen phosphate 1g, sulfur
Acid magnesium (MgSO4·7H2O) 0.5g, agar 20g, rose-bengal solution 3.3mL, the distilled water 1000mL of 1%.121 DEG C of sterilizings
20 minutes, room temperature added 1% streptomycin 3ml when drying in the air to 50-60 DEG C, fully mix, and is poured into culture dish afterwards, standby.Strepto-
The addition of element can suppress the growth of fungus in culture medium effectively.
The determination of fungal culture optimum concentration
Instruct according to Microbiology Experiment and in the past cultivation experience primarily determined that optimum concentration that fungal culture observes was 10-2-
10-3Between.Take the matrix soil in 0.5g matched group to pour in the conical flask equipped with 50ml sterilized water, make 10-2The soil of concentration
Earth leachate, sealing is placed on shaking table, vibrates 20 min with 150 rpm.Vibration stands 10 S after stopping.Draw l mL
Soil extract moves in test tube and adds 9 mL sterilized water, is diluted 10 times, makes 10-3The soil extract of concentration, fills
Divide mixing.Take 200ul soil extract even spread respectively to LB culture medium, culture dish is inverted into the constant temperature training of 28 degree
Supporting in case, lucifuge is cultivated.Continuous Observation fungus growing state, counts after its suitable size quantity is stable.Preliminary experiment
Found that 10-2This concentration is the optimum concentration that in the composting substrate of lawn, fungal culture is observed.When incubation time is two days
Suitable counting.
Fungus plate count method
Experiment uses the fungus in dilution plate rubbing method isolation medium, and specific operation process is: weigh 0.5 g lawn compost base
Matter is poured in the conical flask equipped with 50 mL sterilized water, is placed on shaking table conical flask sealing with 150 rpm vibrations 20
Min, obtains 10-2The soil suspension of concentration.The earth suspension 200ul that fetches earth is expelled on solid rose bengal medium and is coated with
Cloth is uniform, cultivates 2d in culture dish is inverted in 28 DEG C of constant incubators afterwards, selects clump count in the range of 30-300
Flat board carry out separate counting, and press colony characteristics sort out, number.With plate streak, the fungus being separated to is purified,
It is placed in 4 DEG C of refrigerators saving backup.
The molecular biological variety identification method of fungus
The fungus obtaining every kind of pure culture in culture dish utilizes Ezup pillar genome DNA extraction test kit (the raw work in Shanghai) to carry
Take fungal genomic DNA, test by test kit operating process.Gained DNA is used for fungus 18SrDNA PCR amplification real
Test.Amplimer
ITS1 (5 '-TCCGTAGGTGAACCTGCGG-3 '), ITS4 (5 '-TCCTCCGCTTATTGATATGC-3 '),
PCR reaction condition is: 94 DEG C of denaturations 2min, 94 DEG C of degeneration 30s, and 59 DEG C of annealing 30s, 72 DEG C extend 90s, and totally 35 are followed
Ring, last 72 DEG C extend 7min[100].Amplification system is following (50 μ L): primer 1 (27F) 2 μ L；Primer 2 (1492R) 2 μ L；
10x Buffer(Mg2+) 5μL；dNTP 1μL；Taq enzyme 1 μ L；DNA profiling 10 μ L；ddH2O 29μL.Amplified production is each
Take 7 L to mix homogeneously with 2 L 6 × Glycerol DNA loading buffer respectively, take 7 L mixed liquor point samples, carry out
1.2% agarose gel electrophoresis, after EB dyeing, observes, it was observed that by institute after bright object band in ultraviolet gel imaging system
Obtain PCR primer and transfer to Beijing Hua Da genome company purification and order-checking.
Fungus 18SrDNA sequence analysis method
Sequencing result is through NCBI Blast (http: ∥ www.ncbi.nlm.nih.gov) comparison analysis, it is thus achieved that similarity is
The 18SrDNA sequence of five high bacterial strains.ClustalX is used to carry out multi-contrast, carry out phylogeny with MEGA software and divide
Analysis, uses Neighbor-join method phylogenetic tree construction, and number (Bootstrap) of bootstrapping is 1000.Homology journey according to sequence
Degree primarily determines that bacterial strain to be identified status on taxonomy.Cultivation is obtained bacterial strain and the type strain in ncbi database
Compare, if similarity reaches more than 96%, then the bacterial strain that can cultivation be obtained and type strain are classified as a genus, also
Just arrive, by cultivating identification of strains, the level belonged to.
Development results is analyzed
The 18SrDNA that can cultivate fungus in 2.1 substrate analyzes
Through cultivation counting and the kind of fungus in the composting substrate of lawn are divided and sorted out, pick out the fungus of different characteristic
Bacterium colony.
By the PCR to different fungus 18SrDNA, obtain a plurality of amplified fragments, and by with DNA Marker's
Position versus, finds that its band meets the feature of fungus, send the result after survey to show as follows:
This experiment obtains 19 ordered sequences altogether, by the sequence that obtains by Blast program and Gen-Bank data base
Knowing that sequence carries out similarity comparison, its classification information, Similarity Percent and GenBank accession number are shown in Table 1.
Table 1 substrate can be cultivated the 18SrDNA sequence similarity analysis of fungus
By sequence similarity analysis find bacterial strain 1 and bacterial strain 12 be Penicillium (Penicillium), bacterial strain 2,7,9,10,
13,15,16,18 be trichoderma (Trichoderma), bacterial strain 3 for chaetomium globosum belong to (Arthrobacter), bacterial strain 4 is little
Mycosphaerella (Leptosphaeria), bacterial strain 5,6,11 be Fusarium (Fusarium), bacterial strain 8,14,17 is Eurotium
(Aspergillus), bacterial strain 19 be the raw red shell Pseudomonas of light color (Bionectria ochroleuca).Above bacterial strain and pattern bacterium
The similarity of strain all reaches more than 96%, and Reliability ratio is higher.
The sequencing result of fungal bacterial strains different for isolated 19 strain forms is carried out Blast analysis and multiple ratio pair, profit
With MEGA5.1 constructing system cladogram based on 18SrDNA homology.
Develop conclusion
This technology binding molecule sequencing technologies can be cultivated fungus monoid in the composting substrate of lawn after carbon nanomaterial carry out adding
Identification and analysis, main research is as follows: isolated 19 fungal strain altogether, shows this 19 fungal strain through 18SrDNA sequencing result
It is belonging respectively to Penicillium(Penicillium), trichoderma(Trichoderma), chaetomium globosum belong to(Arthrobacter)、
Leptosphaeria(Leptosphaeria), Fusarium(Fusarium), Eurotium (Aspergillus), the raw red shell of light color
Pseudomonas(Bionectria ochroleuca).Total number of fungi and diversity indices can be cultivated without significant difference, but add nanometer
After material, the composition of fungus colony but there occurs change.
Accompanying drawing illustrates:
Fig. 1 is that it is carried out the electrophoretogram after 18SrDNA amplification；
Detailed description of the invention:
The present invention is described below by specific embodiment.Unless stated otherwise, technological means used in the present invention is
Method known in those skilled in the art.It addition, embodiment is interpreted as illustrative, and the model of the unrestricted present invention
Enclosing, the spirit and scope of the invention are limited only by the claims that follow.To those skilled in the art, without departing substantially from this
On the premise of bright spirit and scope, the various changes or the change that carry out the material component in these embodiments and consumption also belong to
In protection scope of the present invention.The present invention is raw materials used, reagent is commercially available.
Embodiment 1
(1) experiment material
Experiment soil takes from Tianjin Normal University campus 10 cm topsoil, and fundamental property is: water content 19.4%, pH
7.27, electrical conductivity 2250 S/cm, phosphorus organic, complete, full nitrogen are respectively 52.3,3.75,2.15 g/kg.
Experiment consumer garbage compost is from Tianjin little Dian destructor plant.PH is 7.49, electrical conductivity 2300 S/cm,
Phosphorus organic, complete, full nitrogen are respectively 132,6.81,25.1 g/kg.Heavy metal Cr, Cu, Pb, Zn, Cd content is respectively 703,
341、217、677、5.01 mg/kg。
Being purchased from Qiu Shi carbon black plant, Tianjin for examination nano carbon black (CB), particle diameter is 20 nm, and specific surface area is 1200 m2/ g,
PH value is 7.0, utilizes KMnO before experiment4To its modification.
Graphene (G) microplate is purchased from Nanjing Ji Cang nanosecond science and technology company limited, and structure is black foil shape, microplate size
0.5 um, thickness 5 nm, specific surface area 40 m2/ g, density about 2.25 g/cm3, electrical conductivity 8000 S/m, phosphorus content > 99.5%.
Graphene oxide (GO) is purchased from Suzhou Heng Qiu nanometer company, for black or brownish yellow powder, average thickness 3.4-7
Nm, lamella diameter 10 μm, specific surface area 100 m2/ g, purity > 90%.
CNT (CNT) is purchased from Beijing and wins space high-tech new material technology company limited, diameter 20 nm, length
10um ,-COOH content 1.43%, purity>90wt%, ashes<8wt%, specific surface area>110 m2/ g, conduction: > 102 s/cm.
(2) experimental technique:
Measure soil moisture content after fetching earth immediately, be calculated the ratio of soil dry weight and weight in wet base.Proportionally calculate to obtain dry weight
Wet soil weight corresponding to 1500 g.Claim the wet soil of corresponding weight mixes with 30 g garbage composts, stir, loading internal diameter
In the plastic flowerpot of high 20 cm of 15 cm, fill 15 basins altogether.
Experiment sets 5 process, Graphene (G), graphene oxide (GO), CNT (CNT), modified Nano white carbon black altogether
(CB) weigh the ratio of 1% in dry ground to be added separately in mixed-matrix, with the process without carbon nanomaterial for comparison (CK),
Fully mix after adding carbon nanomaterial, every basin sowing tall fescue seed 5g.During cultivation, temperature is 19 DEG C, and relative humidity is
60%, every day keeps the skin wet to soil, makes soil moisture be maintained at the 70% of water capacity.After cultivating 130d, by deep for 1cm in basin
Degree soil takes out mixing, crosses 60 mesh sieves, and the preservation of-20 DEG C of refrigerator, for microorganism separation and analysis of accounts.
Embodiment 2
(1) experiment material
Experiment soil takes from Tianjin Normal University campus 20 cm topsoil, and fundamental property is: water content 19.4%, pH
7.27, electrical conductivity 2250 S/cm, phosphorus organic, complete, full nitrogen are respectively 52.3,3.75,2.15 g/kg.
Experiment consumer garbage compost is from Tianjin little Dian destructor plant.PH is 7.49, electrical conductivity 2300 S/cm,
Phosphorus organic, complete, full nitrogen are respectively 132,6.81,25.1 g/kg.Heavy metal Cr, Cu, Pb, Zn, Cd content is respectively 703,
341、217、677、5.01 mg/kg。
Being purchased from Qiu Shi carbon black plant, Tianjin for examination nano carbon black (CB), particle diameter is 70 nm, and specific surface area is 1200 m2/ g,
PH value is 7.0, utilizes KMnO before experiment4To its modification.
Graphene (G) microplate is purchased from Nanjing Ji Cang nanosecond science and technology company limited, and structure is black foil shape, microplate size 20
Um, thickness 25 nm, specific surface area 60 m2/ g, density about 2.25 g/cm3, electrical conductivity 10000 S/m, phosphorus content > 99.5%.
Graphene oxide (GO) is purchased from Suzhou Heng Qiu nanometer company, for black or brownish yellow powder, average thickness 7 nm, sheet
Layer diameter 50 μm, specific surface area 100-300 m2/ g, purity > 90%.
CNT (CNT) is purchased from Beijing and wins space high-tech new material technology company limited, diameter 40 nm, length 30
Um ,-COOH content 1.43%, purity>90wt%, ashes<8wt%, specific surface area>110 m2/ g, conduction: > 102 s/cm.
(2) experimental technique:
Measure soil moisture content after fetching earth immediately, be calculated the ratio of soil dry weight and weight in wet base.Proportionally calculate to obtain dry weight
Wet soil weight corresponding to 1500 g.Claim the wet soil of corresponding weight mixes with 30 g garbage composts, stir, loading internal diameter
In the plastic flowerpot of high 20 cm of 15 cm, fill 15 basins altogether.
Experiment sets 5 process, Graphene (G), graphene oxide (GO), CNT (CNT), modified Nano white carbon black altogether
(CB) weigh the ratio of 1% in dry ground to be added separately in mixed-matrix, with the process without carbon nanomaterial for comparison (CK),
Fully mix after adding carbon nanomaterial, every basin sowing tall fescue seed 5g.During cultivation, temperature is 27 DEG C, and relative humidity is
72%, every day keeps the skin wet to soil, makes soil moisture be maintained at the 70% of water capacity.After cultivating 130d, by deep for 5cm in basin
Degree soil takes out mixing, crosses 60 mesh sieves, and the preservation of-20 DEG C of refrigerator, for microorganism separation and analysis of accounts.
SEQUENCE LISTING
<110>Tianjin Normal University
<120>carbon nanomaterial regulation and control lawn composting substrate Fungal identification is used to analyze method
<160> 2
<170> PatentIn version 3.5
<210> 1
<211> 19
<212> DNA
<213>artificial sequence
<400> 1
tccgtaggtg aacctgcgg 19
<210> 2
<211> 20
<212> DNA
<213>artificial sequence
<400> 2
tcctccgctt attgatatgc 20
Claims (4)
1. one kind uses the method that carbon nanomaterial regulation and control lawn composting substrate Fungal identification is analyzed, it is characterised in that by following
Step is carried out:
(1) experiment material
Experiment soil takes 0-20 cm topsoil；
Being 20 ~ 70 nm for examination nano carbon black particle diameter, specific surface area is 1200 m2/ g, pH value is 7.0, utilizes KMnO before experiment4
To its modification；
Graphene microchip size 0.5-20 um, thickness 5-25 nm, specific surface area 40-60 m2/ g, density about 2.25 g/cm3, electricity
Conductance 8000-10000 S/m, phosphorus content > 99.5%；
Graphene oxide average thickness 3.4-7 nm, lamella diameter 10-50 μm, specific surface area 100-300 m2/ g, purity > 90%；
Carbon nanotube diameter 20-40 nm, length 10-30 um ,-COOH content 1.43%, purity>90wt%, ashes<8wt%, than
Surface area > 110 m2/ g, conduction: > 102 s/cm；
(2) experimental technique:
Measure soil moisture content after fetching earth immediately, be calculated the ratio of soil dry weight and weight in wet base, proportionally calculate to obtain dry weight
Wet soil weight corresponding to 1500 g, claim the wet soil of corresponding weight mixes with 30 g garbage composts, stir, loading internal diameter
In the plastic flowerpot of high 20 cm of 15 cm, fill 15 basins altogether；
Experiment sets 5 process altogether: Graphene (G), graphene oxide (GO), CNT (CNT), modified Nano white carbon black (CB) are pressed
Dry ground weighs the ratio of 1% and is separately added into, and fully mixes, be right with the process without carbon nanomaterial after adding carbon nanomaterial
According to (CK), every basin sowing tall fescue seed 5g, during cultivation, temperature is 19 ~ 27 DEG C, and relative humidity is 60 ~ 72%, and every day is given
Soil keeps the skin wet, and makes soil moisture be maintained at the 70% of water capacity, after cultivating 130d, is taken by 0-5cm deep soil in basin
Going out mixing, cross 60 mesh sieves, the preservation of-20 DEG C of refrigerator, for microorganism separation and analysis of accounts.
2. the method described in claim 1, wherein the fundamental property of soil is: water content 19.4%, pH 7.27, electrical conductivity 2250
S/cm, phosphorus organic, complete, full nitrogen are respectively 52.3,3.75,2.15 g/kg；
Experiment consumer garbage compost is from Tianjin little Dian destructor plant, and pH is 7.49, and electrical conductivity 2300 S/cm is organic
Matter, full phosphorus, full nitrogen are respectively 132,6.81,25.1 g/kg;
Heavy metal Cr, Cu, Pb, Zn, Cd content are respectively 703,341,217,677,5.01 mg/kg；
Nano carbon black, particle diameter is 20 ~ 70 nm, and specific surface area is 1200 m2/ g, pH value is 7.0, utilizes KMnO before experiment4Right
Its modification；
Graphene microchip size 0.5-20 um, thickness 5-25 nm, specific surface area 40-60 m2/ g, density about 2.25 g/cm3, electricity
Conductance 8000-10000 S/m, phosphorus content > 99.5%；
Graphene oxide average thickness 3.4-7 nm, lamella diameter 10-50 μm, specific surface area 100-300 m2/ g, purity > 90%；
CNT, diameter 20-40 nm, length 10-30 um ,-COOH content 1.43%, purity>90wt%, ashes<8wt%, than
Surface area > 110 m2/ g, conduction: > 102 s/cm.
3. use carbon nanomaterial regulation and control lawn composting substrate Fungal identification to analyze method described in claim 1 and determine fungus bacterium
Application in terms of the composition fallen.
4. the application described in claim 3, the composition of wherein said fungus colony includes: Penicillium(Penicillium), wood
Mould Pseudomonas(Trichoderma), chaetomium globosum belong to(Arthrobacter), Leptosphaeria(Leptosphaeria), Fusarium spp.
Belong to(Fusarium), Eurotium (Aspergillus), the raw red shell Pseudomonas of light color(Bionectria ochroleuca)。
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103814744A (en) * | 2014-03-11 | 2014-05-28 | 天津师范大学 | Regulation and control method for improving diversity of nanocarbon to soil nematodes |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103814744A (en) * | 2014-03-11 | 2014-05-28 | 天津师范大学 | Regulation and control method for improving diversity of nanocarbon to soil nematodes |
Non-Patent Citations (3)
| Title |
|---|
| 刘文娟: ""纳米胶囊型农药和碳纳米材料对土壤微生物的生物效应"", 《中国博士学位论文全文数据库 工程科技Ⅰ辑》 * |
| 刘玉真: ""改性纳米黑碳的土壤环境行为及其环境效应研究"", 《中国博士学位论文全文数据库 工程科技Ⅰ辑》 * |
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