CN110511880A - A kind of resistance to uranium bacterium and its method for degradation treatment heavy metal accumulation plant - Google Patents
A kind of resistance to uranium bacterium and its method for degradation treatment heavy metal accumulation plant Download PDFInfo
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Abstract
The invention discloses a kind of resistance to uranium bacterium and its for the method for degradation treatment heavy metal accumulation plant, the bacterium is atrophy bacillus, LatinBacillus atrophaeus, it is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center CGMCC, deposit number 16080, preservation day 20180725.Resistance to uranium bacterium has stronger tolerance, the bad conditions such as resistant against high temperatures, radiation to extreme environment；Biodegrading process is simple, and removal rate is high.
Description
Technical field
The invention belongs to field of biotechnology, and in particular to a kind of resistance to uranium bacterium and its be used for degradation treatment heavy metal accumulation
The method of plant.
Background technique
Heavy metal material is always a kind of indispensable resource in the development of human industry's technology.With getting over now
Carry out the formation of more new high-tech industries, although producing various heavy alternative materials, the exploitation of heavy metal resources
Amount and demand are not reduced, and the exploitation of mankind's counterweight metallic mineral resources greatly increases instead.It is utilized in heavy metal resources
During, including mining area exploitation, tailing are stacked and ore refines and following process is dense using will cause local heavy metal
Height is spent, to have an impact to ambient enviroment, Polluted area can be gradually expanded with rainwater and basin.
According to incompletely statistics, before and after 2010, China's urban industry wastewater emission amount is up to 40,000,000,000 t/, wherein arranging
Strong toxic heavy metal out, such as Cd, Hg have reached 2700t or so, cause about 90% urban river, 75% lake,
50% underground water and 20% arable land receive different degrees of pollution.
The main source of heavy metal pollution is as caused by human activity, comprising: the generation of industrial refuse, fertilizer and pesticide
Abuse, sewage irrigation and atmospheric sedimentation etc..For heavy metal with its special chemical characteristic, it is lasting to have to the pollution of environment, biology
Property and bio-toxicity.Heavy metal can be directly changed soil water body chemical composition, directly or indirectly destroy ecologic structure, and pass through soil
Earth water body-crop system migration accumulation, and then influence human health.
With the development of present industrial and the utilization of nuclear energy, lead to a large amount of exploitations of uranium ore and various metallic ores, thus
Cause a large amount of heavy metal pollution.The exploitation and smelting of metallic ore are to cause the most important reason of Heavy-metal Polluted Environment, because
A large amount of heavy metal tailing and waste material can be generated during this, causes the pollution of water body and ground surface environment, and then influence ecology
Variation.Uranium is polluted, main uranium contaminated site specifically includes that ore extraction and smelts place, retired core in the world
Power station, nuclear weapon test field etc..The toxicity of uranium shows as chemical toxicity and radiotoxicity, and the lower α of the main emitted energy of uranium is penetrated
Line is to cause interior radiation.The long half time of uranium reaches more than one hundred million years, if not administered properly by the place that uranium pollutes, to the mankind and can move
The health of object generates potential, long-term harm, it is therefore necessary to repair by uranium contaminated land.In addition, uranium meeting association is a variety of
Heavy metal, such as lead, strontium, cadmium.Therefore, uranium Polluted area is mostly Compound Heavy Metals, so that pollution amelioration complex.
Although conventional physics and chemical method can be used to handle the heavy metal in water-soil medium, because its is at high cost,
Secondary pollution is easily caused, treatment process harm to the human body is big, thus is not easy to operate on the spot；It compares, it is biological prosthetic to show
More advantages.So far, the biological prosthetic research of Heavy-metal Polluted Environment is confined to carry out in laboratory more, only a small number of real
It tests and enters the crop field stage.
Phytoremediation technology is the research hotspot that field is repaired in current heavy metal pollution, however the weight in relation to generating after repairing
The research of metal enrichment plant disposal technology is opposite to be lacked.
But a large amount of heavy metal accumulation biomass can be generated using enriching plant repairing heavy metal pollution, if disposition is not
When can then cause secondary pollution.Blaylock etc. strengthens Indian mustard (Brassica juncea) using chelating agent EDTA and repairs
Pb contaminated soil, as a result, it has been found that overground part dry weight can be harvested and be up to 6t/hm for every 6 weeks2, the content of Pb is up to 10000-
15000mg/kg.The research such as Chen ciliate desert-grass (Pteris vittata L.) can be used as the super enrichment plant for repairing As contaminated soil
Object, fresh weight is up to 36 t/hm2.Important composition of the heavy metal accumulation biomass disposal technology as plant extract recovery technique
Part, the short slab of the technology are considered as the one of the major reasons for restricting phytoremediation engineering and commercial applications.Therefore, weight
Metal enrichment biomass safe disposal technology is one of the important scientific issues of phytoremediation technology urgent need to resolve from now on.
Component in plant is all largely cellulose, hemicellulose and lignin, in general biomass, cellulose
Ingredient is about between 40%-50%, and hemicellulose accounts for about 10%-30% and lignin is 20%-30%.And these objects
Matter is difficult to the utilization that is degraded by microorganisms again, wherein be most difficult to be utilized by microorganism is lignin.Utmostly, it most effectively degrades
Lignocellulosic is required at present to the efficient degradation for realizing biomass and a large amount volume reduction.
Summary of the invention
In view of the above-mentioned problems, the present invention provides a kind of resistance to uranium bacterium and its side for degradation treatment heavy metal accumulation plant
Method, resistance to uranium bacterium have stronger tolerance, the bad conditions such as resistant against high temperatures, radiation to extreme environment；Biodegrading process is simple,
Removal rate is high.
A kind of resistance to uranium bacterium for solving the above technical problem and its method for degradation treatment heavy metal accumulation plant,
Be characterized in that: the bacterium is bacillus subtilis, and Latin Bacillus subtilis is preserved in Chinese microorganism strain
Preservation administration committee common micro-organisms center CGMCC, deposit number 16080, preservation day 20180711, preservation address: Beijing
The institute 3 of city, North Star West Road, Chaoyang District 1.
The bacillus subtilis, original name Bacillus subtilis endophyticus (Bacillus subtilis
Subsp.Subtilis), there are stronger or strong tolerance, the bad conditions such as resistant against high temperatures, radiation to extreme environment.
The resistance to uranium bacterium is isolated from Uranium tailings；Slag sample is taken in uranium tailings pond, the educable dominant strain of picking,
With the liquid TGY Screening of Media that U (VI) concentration is 20-100mg/L and step by step mutagenesis is tamed, is identified as atrophy gemma bar
Bacterium.
The TGY culture medium prescription includes 1 part of glucose of mass parts, and 5 parts of tryptone, 3 parts of yeast powder, distilled water
1000 parts, adjust pH to 7.0-7.2.The growth temperature of resistance to arsenic bacterium is 35 DEG C.
The process of the strain domestication, comprising the following steps:
(1) preparation of bacteria suspension: logarithmic phase seed liquor is made in the bacterial strain for having cultivated 20h and prepares mutagenesis.
(2) ultraviolet mutagenesis: by shading around superclean bench before experiment, make light wave by ultraviolet light irradiation 30min when
Between after be in stable state, in empty plate be added 4mL culture to 20h bacterium solution and by it in the position apart from ultraviolet lamp 24cm
The irradiation for giving 50min, at regular intervals rotates plate.
(3) the spore suspension gradient dilution for finishing mutagenic treatment in gnotobasis is to 10-4,10-5,10-6 in respectively
Containing 20,50, be coated on the Congo red screening and culturing medium plate of 100mg U, under conditions of 35 DEG C (constant temperature) to by brown paper and
The bacterium solution that black polybag is handled well is cultivated after carrying out mark, and the bacterial strain filtered out is dominant strain.
The method of degradation treatment heavy metal accumulation plant in the present invention, comprising the following steps:
(1) heavy metal accumulation plant steam sterilizing；
(2) sterile solution of urea is added, while adding the bacteria suspension for the antimicrobial composition that inoculum concentration is 10%；
(3) it degrades, the time 30 days.
In the degradation process, 25-40 DEG C of temperature, initial pH value 6-7.5, enriching plant intercept length 1-2.5cm, initially
Water content 40-70%, urea additive amount 1-2.5g/kg；40 DEG C of temperature in prioritization scheme, initial pH value 7.5, enriching plant intercept
Length 1.5cm, initial water content 70%, urea additive amount 2.5g/kg.
The degradation process is reacted in pilot scale degraded pond, and pilot scale degraded pond includes being equipped with degraded pond, ventilation pipe, air
Pump, baffle and ventilation opening, be in degraded pond it is hollow, baffle is located in degraded pond and close to degraded pond bottom surface；Ventilation shaft is fixed
In baffle one side, ventilation opening is set to degraded pond top and close to baffle side；Air pump is set to outside degraded pond, and ventilation pipe passes through
Ventilation opening is connect with air pump；The length of pilot scale degraded pond described in prioritization scheme is respectively 480mm, 475mm, 225 mm；
Its volume is 53.956dm3；Advanced optimize the grid for being also equipped with that one layer of aperture side length is 1mm on baffle described in scheme
Item, to prevent experimental material from entering bottom reserved space from ventilation opening；Baffle described in prioritization scheme is from degraded pond bottom surface 15mm
Place, reserved space account for the 6.3% of degraded pond total volume.
Detailed description of the invention
With reference to the accompanying drawing and specific embodiment does more detailed description to the present invention:
Fig. 1 is growth curve chart of the bacterium without U environment
Fig. 2 is growth curve chart of the bacterium under 100mg/LU culture
Fig. 3 is total degradation rate figure of the bacterial strain to enrichment rye grass
Fig. 4 is bacterial strain respectively to the degradation efficiency figure of cellulose, hemicellulose and lignin in enrichment rye grass
Wherein, CK is control group, and A is atrophy bacillus
Specific embodiment
Invention is further explained With reference to embodiment:
Embodiment 1
A kind of resistance to uranium bacterium, bacterium are bacillus subtilis also known as atrophy bacillus, Latin Bacillus
Subtilis, is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center CGMCC, and deposit number is
16080, preservation day 20180711, preservation address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3.
Bacillus subtilis, original name Bacillus subtilis endophyticus (Bacillus subtilis
Subsp.Subtilis), there are stronger or strong tolerance, the bad conditions such as resistant against high temperatures, radiation to extreme environment.
Resistance to uranium bacterium is isolated from Uranium tailings；Slag sample is taken in uranium tailings pond, the educable dominant strain of picking uses U
(VI) concentration is the liquid TGY Screening of Media of 20mg/L and tames mutagenesis step by step, is identified as atrophy bacillus.
TGY culture medium prescription includes 1 part of glucose of mass parts, and 5 parts of tryptone, 3 parts of yeast powder, distilled water 1000
Part, adjust pH to 7.0.The growth temperature of resistance to arsenic bacterium is 35 DEG C.
The process of strain domestication, comprising the following steps:
(1) preparation of bacteria suspension: logarithmic phase seed liquor is made in the bacterial strain for having cultivated 20h and prepares mutagenesis.
(2) ultraviolet mutagenesis: by shading around superclean bench before experiment, make light wave by ultraviolet light irradiation 30min when
Between after be in stable state, in empty plate be added 4mL culture to 20h bacterium solution and by it in the position apart from ultraviolet lamp 24cm
The irradiation for giving 50min, at regular intervals rotates plate.
(3) the spore suspension gradient dilution for finishing mutagenic treatment in gnotobasis is to 10-4,10-5,10-6 in respectively
Containing 20,50, be coated on the Congo red screening and culturing medium plate of 100mg U, under conditions of 35 DEG C (constant temperature) to by brown paper and
The bacterium solution that black polybag is handled well is cultivated after carrying out mark, and the bacterial strain filtered out is dominant strain.
Embodiment 2
Content in other contents such as embodiment 1, resistance to uranium bacterium are isolated from Uranium tailings；Slag sample is taken in uranium tailings pond,
The educable dominant strain of picking with the liquid TGY Screening of Media that U (VI) concentration is 60mg/L and step by step tames mutagenesis, warp
It is accredited as atrophy bacillus.
TGY culture medium prescription includes 1 part of glucose of mass parts, and 5 parts of tryptone, 3 parts of yeast powder, distilled water 1000
Part, pH to 7.1 is adjusted, the growth temperature of resistance to arsenic bacterium is 35 DEG C.
The process of strain domestication, comprising the following steps:
(1) preparation of bacteria suspension: logarithmic phase seed liquor is made in the bacterial strain for having cultivated 20h and prepares mutagenesis.
(2) ultraviolet mutagenesis: by shading around superclean bench before experiment, make light wave by ultraviolet light irradiation 30min when
Between after be in stable state, in empty plate be added 4mL culture to 20h bacterium solution and by it in the position apart from ultraviolet lamp 24cm
The irradiation for giving 50min, at regular intervals rotates plate.
(3) the spore suspension gradient dilution for finishing mutagenic treatment in gnotobasis is to 10-4,10-5,10-6 in respectively
Containing 20,50, be coated on the Congo red screening and culturing medium plate of 100mg U, under conditions of 35 DEG C (constant temperature) to by brown paper and
The bacterium solution that black polybag is handled well is cultivated after carrying out mark, and the bacterial strain filtered out is dominant strain.
Embodiment 3
Content in other contents such as embodiment 1, resistance to uranium bacterium are isolated from Uranium tailings；Slag sample is taken in uranium tailings pond,
The educable dominant strain of picking with the liquid TGY Screening of Media that U (VI) concentration is 100mg/L and step by step tames mutagenesis, warp
It is accredited as atrophy bacillus.
TGY culture medium prescription includes 1 part of glucose of mass parts, and 5 parts of tryptone, 3 parts of yeast powder, distilled water 1000
Part, adjust pH to 7.2.The growth temperature of resistance to arsenic bacterium is 35 DEG C.
The process of strain domestication, comprising the following steps:
(1) preparation of bacteria suspension: logarithmic phase seed liquor is made in the bacterial strain for having cultivated 20h and prepares mutagenesis.
(2) ultraviolet mutagenesis: by shading around superclean bench before experiment, make light wave by ultraviolet light irradiation 30min when
Between after be in stable state, in empty plate be added 4mL culture to 20h bacterium solution and by it in the position apart from ultraviolet lamp 24cm
The irradiation for giving 50min, at regular intervals rotates plate.
(3) the spore suspension gradient dilution for finishing mutagenic treatment in gnotobasis is to 10-4,10-5,10-6 in respectively
Containing 20,50, be coated on the Congo red screening and culturing medium plate of 100mg U, under conditions of 35 DEG C (constant temperature) to by brown paper and
The bacterium solution that black polybag is handled well is cultivated after carrying out mark, and the bacterial strain filtered out is dominant strain.
Embodiment 4
The method of degradation treatment heavy metal accumulation plant, comprising the following steps:
(1) heavy metal accumulation plant steam sterilizing；
(2) sterile solution of urea is added, while adding the bacteria suspension for the antimicrobial composition that inoculum concentration is 10%；
(3) it degrades, the time 30 days.
In degradation process, 25 DEG C of temperature, initial pH value 6, enriching plant intercept length 1cm, initial water content 40%, urea
Additive amount 1g/kg；
Embodiment 5
The method of degradation treatment heavy metal accumulation plant, comprising the following steps:
(1) heavy metal accumulation plant steam sterilizing；
(2) sterile solution of urea is added, while adding the bacteria suspension for the antimicrobial composition that inoculum concentration is 10%；
(3) it degrades, the time 30 days.
In degradation process, 40 DEG C of temperature, initial pH value 7.5, enriching plant intercept length 2.5cm, initial water content
70%, urea additive amount 2.5g/kg；
Embodiment 6
The method of degradation treatment heavy metal accumulation plant, comprising the following steps:
(1) heavy metal accumulation plant steam sterilizing；
(2) sterile solution of urea is added, while adding the bacteria suspension for the antimicrobial composition that inoculum concentration is 10%；
(3) it degrades, the time 30 days.
In degradation process, 40 DEG C of temperature, initial pH value 7.5, enriching plant intercept length 1.5cm, initial water content
70%, urea additive amount 2.5g/kg.
Embodiment 7
The method of degradation treatment heavy metal accumulation plant, comprising the following steps:
(1) heavy metal accumulation plant steam sterilizing；
(2) sterile solution of urea is added, while adding the bacteria suspension for the antimicrobial composition that inoculum concentration is 10%；
(3) it degrades, the time 30 days.
In degradation process, 30 DEG C of temperature, initial pH value 7, enriching plant intercept length 2cm, initial water content 50%, urea
Additive amount 2g/kg；
Embodiment 9
The method of degradation treatment heavy metal accumulation plant, comprising the following steps:
(1) heavy metal accumulation plant steam sterilizing；
(2) sterile solution of urea is added, while adding the bacteria suspension for the antimicrobial composition that inoculum concentration is 10%；
(3) it degrades, the time 30 days.
In degradation process, 35 DEG C of temperature, initial pH value 6.5, enriching plant intercept length 1.5cm, initial water content
60%, urea additive amount 1.5g/kg；
Embodiment 8
The method of degradation treatment heavy metal accumulation plant, comprising the following steps:
(1) heavy metal accumulation plant steam sterilizing；
(2) sterile solution of urea is added, while adding the bacteria suspension for the antimicrobial composition that inoculum concentration is 10%；
(3) it degrades, the time 30 days.
In degradation process, 40 DEG C of temperature, initial pH value 7.5, enriching plant intercept length 1.5cm, initial water content 70%,
Urea additive amount 2.5g/kg.
Degradation process is reacted in pilot scale degraded pond, pilot scale degraded pond include equipped with degraded pond, ventilation pipe, air pump,
Baffle and ventilation opening, be in degraded pond it is hollow, baffle is located in degraded pond and close to degraded pond bottom surface；Ventilation shaft is fixed on gear
Plate one side, ventilation opening are set to degraded pond top and close to baffle sides；Air pump is set to outside degraded pond, and ventilation pipe passes through ventilation
Mouth is connect with air pump；
Pilot scale degraded pond length is respectively 480mm, 475mm, 225mm；Its volume is 53.956dm3；It is further excellent
Also be equipped on baffle described in change scheme one layer of aperture side length be 1mm grid item, with prevent experimental material from ventilation opening into
Enter bottom reserved space；Baffle described in prioritization scheme from degraded pond bottom surface 15mm at, reserved space accounts for degraded pond total volume
6.3%.
Test one
Degradation effect of the bacterial strain to uranium enrichment rye grass
Processing method: using 150mL triangular flask for container of degrading, and the heavy metal that 10g intercept length is 1.5cm drying is added
It is enriched with rye grass, the bacteria suspension that 90mL total amount is 10% is added in closing sterilizing, to add distilled water as control group, every group parallel 3
It is secondary, it is placed in 37 DEG C of incubators and cultivates 30d.After the degradation of 30d, degradation residue is taken out, is repeatedly washed on a small quantity with distilled water
It is limpid to cleaning solution to wash degradation residue, degrades cleaning container 3 times, filter paper filtering collects cleaning solution and measures its volume；Residue dries
Do to constant weight, weigh, degraded the weightlessness of front and back rye grass by measurement, be calculated different microorganisms degrade to rye grass it is total
Degradation rate；The content of beary metal for measuring rye grass degradation residue and raffinate by ICP-MS respectively, is calculated different microorganisms
To the leaching rate of heavy metal.The measurement of residue progress cellulose, hemicellulose and content of lignin.Total degradation rate and cellulose,
Following (the measurement side in later tests of hemicellulose, the measurement of the degradation rate of lignin and leaching rate of heavy metals and calculation method
Method is identical):
The measuring method of total degradation rate
According to the weightlessness before and after microbial degradation heavy metal accumulation rye grass, microorganism can be intuitively shown to a huge sum of money
Belong to enrichment rye grass degradation effect, by the above processing method processing after, degradation rate calculation formula such as:
The degradation rate (%) of Dr-- heavy metal accumulation rye grass；M0-- the initial dry biomass of heavy metal accumulation rye grass
(g)；Remaining dry biomass (g) after the degradation of M1-- heavy metal accumulation rye grass.
The measurement of content of cellulose uses anthrone colorimetry, and the measurement of hemicellulose level uses DNS colorimetric method, lignin
The measurement of content uses acetyl bromide colorimetric method.
The calculation method of cellulose, hemicellulose and Lignin degradation rate
It is fine respectively by the measurement of heavy metal accumulation rye grass degradation front and back cellulose, hemicellulose and content of lignin
It is as follows to tie up element, hemicellulose, the degradation rate calculation formula of lignin.
The degradation rate (%) of tri- kinds of substances of Dr--；C0-- the content (g/g) of three kinds of substances of rye grass before degrading；M0Before degradation
Sample quality (g)；C1-- the content (g/g) of three kinds of substances of rye grass after degradation；M1-- sample quality (g) after degradation.
The measuring method of leaching rate of heavy metals
For example above processing method of sample handling processes.Leaching rate of heavy metals calculation formula:
Lr-- leaching rate of heavy metals (%)；C1-- raffinate heavy metal concentration (mg/L) after degradation；V1-- raffinate volume after degradation
(L)；C2-- residue heavy metal concentration (mg/kg) after degradation；M1-- mass of residue (kg) after degradation.
Indices measurement result indicates that each index is respectively provided with 3 repetitions with mean+SD.Using
2003 software of Microsoft Excel carries out data statistics；Single factor test is carried out to determination data with DPS data processing system
Difference degree statisticallys analyze between variance analysis and the processing of LSD multiple comparative test, and orthogonal result uses extremum difference analysis；Using
9.0 software of Origin carries out Plot Work.
It is enriched with rye grass Initial Composition and Analysis of Heavy Metals
The Initial Composition and content of beary metal of rye grass the results are shown in Table 1.
Table 1 is enriched with rye grass Initial Composition and content of beary metal (dry weight)
Test carries out the volume reduction research of 30d degradation by a definite date to enrichment rye grass using microorganism, after enrichment rye grass degradation
Biomass weight-loss ratio can express the degradation volume reduction effect of microorganism；Measurement enrichment rye grass degradation front and back cellulose, hemicellulose
The changes of contents and degradation residue group variation of element and lignin, can analyze different microorganisms to heavy metal accumulation rye grass
The main matter of degradation investigates the degradation volume reduction effect of enrichment rye grass from another point of view；In addition, weight after enrichment rye grass degradation
The leaching rate of metal can effectively show heavy metal from solid biomass to the migration effect of degradation raffinate.
Total degradation rate of the bacterial strain to enrichment rye grass
Uranium is enriched with rye grass after sterilizing, is inoculated with degradation bacteria strains, the weightlessness being enriched with before and after rye grass according to strains for degrading,
Bacterial strain can intuitively be shown to the overall degradation effect of enrichment rye grass, such as Fig. 3.As can be seen from the figure microorganism is to richness
Collection rye grass single bacterium degradation effect overall degradation rate reaches 40% or more.
Bacterial strain is respectively to the degradation effect of the cellulose of enrichment rye grass, hemicellulose and lignin
By the enrichment rye grass degradation process of 30d, bacterial strain is to cellulose, hemicellulose and lignin in enrichment rye grass
Degradation effect such as Fig. 4.From the point of view of the weight-loss ratio of enrichment rye grass, bacterial strain is to the degradation rate of enrichment rye grass in 35%-60%
Between, wherein the degradation rate to hemicellulose reaches 55%.Lignin is a kind of amorphous high polymer of complexity, is difficult sour
It degrades with enzyme.Microorganism is poor compared to the degradation effect of cellulose and hemicellulose to the degradation effect of lignin.
Strains for degrading is enriched with the Leaching of Heavy Metals effect after rye grass
Purpose using microbiological treatment heavy metal accumulation rye grass has two o'clock；First is the degradation energy using microorganism
Power carries out volume reduction treatment to a large amount of enrichment rye grass；Second is the damage capability using microorganism to enrichment rye grass, so that
Heavy metal is separated from rye grass.Free liquid is moved to by biomass to heavy metal to obtain the microbial degradation of enrichment rye grass
The case where state environment, test carry out content of beary metal measurement for the degradation residue and raffinate of enrichment rye grass respectively, obtain micro-
Analysis of the biology to different leaching rate of heavy metals, such as table 2.
Table 2 is enriched with the leaching rate of heavy metals (%) after rye grass microbial degradation
(note: A, atrophy bacillus)
Bacillus subtilis is higher to the leaching rate of U it can be seen from upper table 2, the removal rate of Cr, Cd and Sr be 90% with
On.
Test two
Tolerance of the bacterial strain to uranium
The content of U is relatively high in heavy metal accumulation rye grass, using the 100mg/ being not much different with degradation environment U concentration
L is resistant to U concentration.Bacterium measures increment using spectrophotometry to the tolerance of U, such as Fig. 1 and Fig. 2.
Under the same conditions, the growth stationary phase of bacterium is in 48h or so.It can be seen from the figure that being not added with the control group of U
In, the growing way of bacterium can be with；In 100mg/L U environment, bacterial growth receives certain influence, but still has more good
Good growth vigor.
Basic principles and main features and advantages of the present invention of the invention, above-described embodiment has been shown and described above
It is merely illustrated the principles of the invention with described in specification, without departing from the spirit and scope of the present invention, the present invention
It will also have various changes and improvements, these changes and improvements are fallen in scope of the claimed invention.The present invention claims
The range of protection is defined by the appending claims and its equivalent thereof.
Claims (8)
1. a kind of resistance to uranium bacterium, it is characterised in that: the bacterium is atrophy bacillus, LatinBacillus subtilis
, it is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center CGMCC, deposit number 16080, preservation day
20180711。
2. a kind of resistance to uranium bacterium according to claim 1, it is characterised in that: atrophy bacillus has extreme environment
Strong tolerance, resistant against high temperatures, radiation bad condition.
3. a kind of resistance to uranium bacterium according to claim 1, it is characterised in that: it is isolated from Uranium tailings, in uranium tailings pond
Take slag sample, the educable dominant strain of picking, with the liquid TGY Screening of Media that U (VI) concentration is 20-100 mg/L and by
Grade domestication mutagenesis, is identified as atrophy bacillus.
4. a kind of resistance to uranium bacterium according to claim 3, it is characterised in that: the strain has a process of domestication, including with
Lower step:
(1) preparation of bacteria suspension: logarithmic phase seed liquor is made in the bacterial strain cultivated and prepares mutagenesis；
(2) it ultraviolet mutagenesis: by shading around superclean bench before experiment, is in light wave after ultraviolet light irradiation and stablizes shape
State is added 4 mL culture to the bacterium solution of 20 h in empty plate and gives it to 50 min in the position apart from 24 cm of ultraviolet lamp
Irradiation, plate is rotated at regular intervals；
(3) in gnotobasis by spore suspension gradient dilution that mutagenic treatment finishes to 10-4,10-5,10-6 in containing respectively
20, it 50, is coated on the Congo red screening and culturing medium plate of 100mg U, to by brown paper and black polybag under conditions of constant temperature
The bacterium solution handled well is cultivated after carrying out mark, and the bacterial strain filtered out is dominant strain.
5. a kind of resistance to uranium bacterium according to claim 3, it is characterised in that: the TGY culture medium includes the Portugal of mass parts
1 part of grape sugar, 3 parts of yeast powder, 1000 parts of distilled water, adjusts pH to 7.0-7.2 by 5 parts of tryptone.
6. a kind of method of the bacterial degradation of resistance to uranium processing heavy metal accumulation plant according to claim 7, it is characterised in that:
It is characterized by comprising following steps:
(1) heavy metal accumulation plant steam sterilizing；
(2) sterile solution of urea is added, while adding the bacteria suspension for the antimicrobial composition that inoculum concentration is 10%；
(3) it degrades, the time 30 days.
7. a kind of method of the bacterial degradation of resistance to uranium processing heavy metal accumulation plant according to claim 1, it is characterised in that:
In the degradation process, 25-40 DEG C of temperature, initial pH value 6-7.5, enriching plant intercept length 1-2.5 cm initially contains
Water 40-70%, urea additive amount 1-2.5 g/kg；40 DEG C of temperature in prioritization scheme, initial pH value 7.5, enriching plant intercept are long
Spend 1.5 cm, initial water content 70%, 2.5 g/kg of urea additive amount.
8. a kind of method of the bacterial degradation of resistance to uranium processing heavy metal accumulation plant according to claim 5, it is characterised in that:
The degradation process is reacted in pilot scale degraded pond, and pilot scale degraded pond includes being equipped with degraded pond, ventilation pipe, air pump, baffle
And ventilation opening, be in degraded pond it is hollow, baffle is located in degraded pond and close to degraded pond bottom surface；Ventilation shaft is fixed on baffle one
Side, ventilation opening are set to degraded pond top and close to baffle sides；Air pump be set to degraded pond outside, ventilation pipe pass through ventilation opening with
Air pump connection；The length of pilot scale degraded pond described in prioritization scheme is respectively 480 mm, 475 mm, 225 mm；Its volume
For 53.956 dm3；The grid item for being also equipped with that one layer of aperture side length is 1 mm on baffle described in scheme is advanced optimized, with
Prevent experimental material from entering bottom reserved space from ventilation opening；Baffle described in prioritization scheme from 15 mm of degraded pond bottom surface at, in advance
Spacing accounts for the 6.3% of degraded pond total volume.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110628759A (en) * | 2019-09-23 | 2019-12-31 | 西南科技大学 | Method for screening and enriching new germplasm of plant symbiotic bacteria by mutation of radiation-resistant wild arbuscular mycorrhizal fungi |
| CN111014283A (en) * | 2020-01-14 | 2020-04-17 | 南华大学 | Method for extracting uranium from plants through chemical-biological synergistic enhancement |
| CN112694982A (en) * | 2021-02-08 | 2021-04-23 | 中国科学院合肥物质科学研究院 | Candida utilis and application thereof |
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| CN115181701A (en) * | 2022-07-15 | 2022-10-14 | 有研资源环境技术研究院(北京)有限公司 | Phosphate solubilizing bacteria and U (VI) solidifying method thereof |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100143316A1 (en) * | 2008-12-05 | 2010-06-10 | Taiwan Agricultural Chemicals And Toxic Substances Research Institute, | Novel strain of bacillus amyloliquefaciens and its use |
| US20150147303A1 (en) * | 2008-12-05 | 2015-05-28 | Feng-Chia Hsieh | Novel strain of bacillus amyloliquefaciens and its use |
| WO2016022779A1 (en) * | 2014-08-06 | 2016-02-11 | Envera, Llc | Bacterial spore compositions for industrial uses |
| CN105950480A (en) * | 2016-05-17 | 2016-09-21 | 西南科技大学 | Pasture degradation volume reduction method |
| CN106944473A (en) * | 2017-03-30 | 2017-07-14 | 西南科技大学 | A kind of method that herbage microbial association repairs uranium contaminated soil |
| CN109055263A (en) * | 2018-08-13 | 2018-12-21 | 西南科技大学 | The antimicrobial composition and biodegrading process of degradation heavy metal accumulation plant |
| CN109082383A (en) * | 2018-08-13 | 2018-12-25 | 西南科技大学 | A kind of antimicrobial composition and biodegrading process of uranium enriching plant of degrading |
-
2018
- 2018-08-14 CN CN201810926046.4A patent/CN110511880B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100143316A1 (en) * | 2008-12-05 | 2010-06-10 | Taiwan Agricultural Chemicals And Toxic Substances Research Institute, | Novel strain of bacillus amyloliquefaciens and its use |
| US20150147303A1 (en) * | 2008-12-05 | 2015-05-28 | Feng-Chia Hsieh | Novel strain of bacillus amyloliquefaciens and its use |
| WO2016022779A1 (en) * | 2014-08-06 | 2016-02-11 | Envera, Llc | Bacterial spore compositions for industrial uses |
| CN105950480A (en) * | 2016-05-17 | 2016-09-21 | 西南科技大学 | Pasture degradation volume reduction method |
| CN106944473A (en) * | 2017-03-30 | 2017-07-14 | 西南科技大学 | A kind of method that herbage microbial association repairs uranium contaminated soil |
| CN109055263A (en) * | 2018-08-13 | 2018-12-21 | 西南科技大学 | The antimicrobial composition and biodegrading process of degradation heavy metal accumulation plant |
| CN109082383A (en) * | 2018-08-13 | 2018-12-25 | 西南科技大学 | A kind of antimicrobial composition and biodegrading process of uranium enriching plant of degrading |
Non-Patent Citations (19)
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110628759A (en) * | 2019-09-23 | 2019-12-31 | 西南科技大学 | Method for screening and enriching new germplasm of plant symbiotic bacteria by mutation of radiation-resistant wild arbuscular mycorrhizal fungi |
| CN111014283A (en) * | 2020-01-14 | 2020-04-17 | 南华大学 | Method for extracting uranium from plants through chemical-biological synergistic enhancement |
| CN112694982A (en) * | 2021-02-08 | 2021-04-23 | 中国科学院合肥物质科学研究院 | Candida utilis and application thereof |
| CN112694982B (en) * | 2021-02-08 | 2022-08-12 | 中国科学院合肥物质科学研究院 | Candida utilis and application thereof |
| CN113426828A (en) * | 2021-07-30 | 2021-09-24 | 西南科技大学 | Method for restoring strontium-polluted soil |
| CN113426828B (en) * | 2021-07-30 | 2022-11-11 | 西南科技大学 | Method for restoring strontium-polluted soil |
| CN115181701A (en) * | 2022-07-15 | 2022-10-14 | 有研资源环境技术研究院(北京)有限公司 | Phosphate solubilizing bacteria and U (VI) solidifying method thereof |
| CN115181701B (en) * | 2022-07-15 | 2023-10-27 | 有研资源环境技术研究院(北京)有限公司 | Phosphate-solubilizing bacterium and method for solidifying U (VI) by using same |
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