CN109679863A - A kind of oil-polluted soils microorganism remediation reagent and preparation method thereof - Google Patents
A kind of oil-polluted soils microorganism remediation reagent and preparation method thereof Download PDFInfo
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- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
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Abstract
The invention discloses a kind of oil-polluted soils microorganism remediation reagents and preparation method thereof, including the following raw material: microorganism reagent: physics regulator: auxiliary material agent is 3.8-7L: 1.6-3kg: 1.5-2.8kg；Microorganism reagent includes following parts by volume: 10-15 parts of black-koji mould, 10-20 parts of pseudomonad, 10-15 parts of candida utili bacterium, 8-10 parts of sticky red rhodotorula；Living bacteria count is hundred million/milliliter of 1-2 in microorganism reagent；Physics regulator includes following raw material: 8-15 parts of charcoal, 8-15 parts of dolomite dust；Auxiliary material agent includes following raw material: 10-20 parts of sophorolipid, 2-3 parts of urea, 3-5 parts of glucose.The preparation method comprises the following steps: each microorganism fungus kind is activated respectively, is spread cultivation, urea is added after mixing and glucose remixes, stationary culture, dolomite and charcoal of the addition through high-temperature sterilization add sophorolipid and be uniformly mixed, and obtain finished product.The present invention uses specified raw material and specific preparation method, and the product of acquisition can safely and effectively degrade oil pollution in soil, improvement soil environment.
Description
Technical field
The present invention relates to technical field of biological remediation, more particularly to a kind of oil-polluted soils microorganism remediation
Reagent and preparation method thereof.
Background technique
The energy and industrial chemicals of the petroleum as a kind of preciousness, its exploitation and utilization have people's production and life
Important influence.But in exploitation and transportational process, can inevitably enter in the soil around oil field because of a variety of causes,
After it enters soil, the serious contaminated soil environment of meeting destroys soil texture, influences the permeability of soil, and will affect plant
Growth.
Therefore, how to provide it is a kind of rapidly and efficiently, the reagent of the remedying oil-polluted soils of safety and environmental protection be this field skill
The problem of art personnel's urgent need to resolve.
Summary of the invention
In view of this, the present invention provides a kind of oil-polluted soils microorganism remediation reagents and preparation method thereof.
To achieve the goals above, the present invention adopts the following technical scheme:
A kind of oil-polluted soils microorganism remediation reagent, including the following raw material: microorganism reagent, physics adjust reagent,
Auxiliary material reagent；
Microorganism reagent parts by volume: physics adjusts reagent parts by weight: auxiliary material reagent weight ratio is 3.8-7L: 1.6-
3kg:1.5-2.8kg；
Microorganism reagent includes the strain of following parts by volume: 10-15 parts of black-koji mould, 10-20 parts of pseudomonad produce protein vacation
Silk 10-15 parts of saccharomycete, 8-10 parts of sticky red rhodotorula；
The living bacteria count of any strain is hundred million/milliliter of 1-2 in microorganism reagent；
It includes following raw material that physics, which adjusts reagent: 8-15 parts of charcoal, 8-15 parts of dolomite dust；
Auxiliary material reagent includes the raw material of following parts by weight: 10-20 parts of sophorolipid, 2-3 parts of urea, 3-5 parts of glucose.
The beneficial effects of the present invention are: firstly, using the black-koji mould of special ratios, pseudomonad, production protein in the present invention
Candidiasis, sticky red rhodotorula can give full play to the collaboration between each bacterial strain as major microorganisms renovation agent raw material
Effect, promotes the growth and breeding of each strain, and the petroleum in the soil that can effectively degrade, and repairs by the soil of oil pollution
Environment；It also added sophorolipid, urea, glucose as auxiliary material reagent in the present invention simultaneously, urea and glucose can be micro-
Biology provides long-term nutritional ingredient, promotes the growth and breeding of microorganism, and sophorolipid can have as biosurfactant
The degradation for promoting the petroleum pollution in soil of effect；And charcoal and dolomite are additionally used in the present invention as physical modulated
Reagent is saved, wherein charcoal can adsorb the petroleum pollution in soil, also under conditions of not influencing microorganism vegetative activity
Can reduce the soil weight, increase soil aeration amount, increase soil moisture content, increase soil adsorption capacity and exchange activity,
The utilization rate for increasing nutrient, can also improve acid soil；And calcium and magnesium rich in dolomite, can not only it improve
The acidity of soil can also promote mineralization, improve soil and absorb and release effect to nutrient, and crop can also be promoted raw
It is long, improve crop yield.And the present invention is using based on microorganism remediation, it is physically improved supplemented by restorative procedure, no nuisance
Matter residual, oil-polluted soils microorganism remediation reagent repairing effect of the present invention is good, safety and environmental protection.
Further, pseudomonad is pseudomonas aeruginosa.
Further, the partial size of dolomite dust is 80-100 mesh.
Further, the partial size of charcoal is 60-80 mesh.
The invention also discloses a kind of preparation methods of oil-polluted soils microorganism remediation reagent, include the following steps:
Step (1): the physical modulated is weighed according to the weight ratio of above-mentioned oil-polluted soils microorganism remediation reagent
Save reagent and the auxiliary material reagent；
Step (2): black-koji mould, pseudomonad, candida utili bacterium, sticky red rhodotorula are activated respectively, expanded
Training；
Step (3): the strain liquid of each strain obtained after spreading cultivation is according to above-mentioned oil-polluted soils microorganism remediation
The parts by volume ratio of reagent is uniformly mixed, and obtains microorganism mixed liquor；
Step (4): the microorganism mixed liquor that step (3) are obtained is uniformly mixed, and urea is added and glucose is uniformly mixed,
Stationary culture 10-15h obtains microorganism remediation liquid；
Step (5): dolomite and charcoal are uniformly mixed, and through high-temperature sterilization, are cooled to room temperature, and are obtained physics and are adjusted examination
Agent finished product；
Step (6): by physics described in microorganism remediation liquid described in step (4) and step (5) adjust reagent finished product with
And auxiliary material reagent sophorolipid is uniformly mixed, and obtains oil-polluted soils microorganism remediation reagent finished product.
The beneficial effect of preparation method of the present invention is: the present invention is first activated and is cultivated to each bacterial strain, can be promoted
Bacterial strain increases rapidly, then obtains microorganism remediation liquid after being mixed, and can effectively improve the adaptability of bacterial strain, then
Microorganism remediation liquid and physics are adjusted into reagent finished product again and auxiliary material reagent mixes, it is ensured that bacterial strain is having powerful fit
Should and fertility, can more effectively play the synergistic effect between each raw material, enhance its effect.
Further, the activation step of black-koji mould described in step (1) are as follows: aspergillus niger is connected to improvement Martin's culture medium and is shaken
In bottle, shaken cultivation about 5-8 days under 25~30 DEG C of cultures；
The step that spreads cultivation of black-koji mould are as follows: the bacterium solution after activation is crossed on improvement Martin's agar slant, 25~30
DEG C stationary culture 5-7 days.
Further, in step (1) pseudomonad culture medium prescription are as follows: gelatin freeze: 15-18g/L, peptone 8-9g/L,
K2SO412-14g/L, MgCL21.5-1.6g/L, agar 16-20g/L, cetrimonium bronmide 03-0.4g/L；PH is 6-
8；
Specific training method are as follows: pseudomonad is inoculated on the culture medium, in 35-37 DEG C of culture 49-52h.
Further, candida utili bacterium described in step (1): activation medium glucose 20g/L, peptone 20g/L,
Yeast powder 10g/L, the water of surplus；
Minimal medium: glucose 30g/L, (NH4)2SO45g/L, yeast powder 3g/L, MgSO40.4g/L, K2HPO41.0g/
L, the water of surplus；
Specific training method: being first inoculated on activation medium, at 27-28 DEG C, sways culture 15-20h；
Again on minimal medium, continue stationary culture 24-26h at 30-32 DEG C.
Further, in step (1) sticky red rhodotorula culture medium are as follows: concentration of glucose 15g/L, yeast extract concentration be 2-
3g/L, malt extract concentration are 3-5g/L, and ammonium sulfate concentrations 2-3g/L, biphosphate potassium concn is 0.5-0.8g/L, seven hydrations
Magnesium sulfate concentration is 0.26-0.28g/L, the water of surplus；PH is 6-8；
Specific training method are as follows: stationary culture 6-9 days at 28-32 DEG C.
Further, high-temperature sterilization temperature described in step (4) is 130-150 DEG C.
It can be seen via above technical scheme that compared with prior art, the present disclosure provides a kind of oil pollution soil
Earth microorganism remediation reagent and preparation method thereof, the present invention use the microorganism of specific proportion, give full play to the kind of each bacterial strain
The interior synergistic effect between population, and the peripheral doses effect of charcoal, dolomite is combined, then using special component and match
The auxiliary reagent of ratio obtains efficient, safety and environmental protection oil-polluted soils microorganism remediation reagent, and preparation method of the present invention letter
It is single, low in cost.
Specific embodiment
The following is a clear and complete description of the technical scheme in the embodiments of the invention, it is clear that described embodiment
Only a part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention, the common skill in this field
Art personnel every other embodiment obtained without making creative work belongs to the model that the present invention protects
It encloses.
The embodiment of the invention discloses oil-polluted soils microorganism remediation reagents, including the following raw material: microorganism reagent,
Physics adjusts reagent, auxiliary material reagent；
Microorganism reagent parts by volume: physics adjusts reagent parts by weight: auxiliary material reagent weight ratio is 3.8-7L: 1.6-
3kg:1.5-2.8kg；
Microorganism reagent includes the strain of following parts by volume: 10-15 parts of black-koji mould, 10-20 parts of pseudomonad produce protein vacation
Silk 10-15 parts of saccharomycete, 8-10 parts of sticky red rhodotorula；
The living bacteria count of any strain is hundred million/milliliter of 1-2 in microorganism reagent；
It includes following raw material that physics, which adjusts reagent: 8-15 parts of charcoal, 8-15 parts of dolomite dust；
Auxiliary material reagent includes the raw material of following parts by weight: 10-20 parts of sophorolipid, 2-3 parts of urea, 3-5 parts of glucose.
Black-koji mould, pseudomonad, candida utili bacterium, sticky red rhodotorula are lived respectively in the embodiment of the present invention
Change, spread cultivation that the specific method is as follows:
The activation step of black-koji mould are as follows: aspergillus niger is connected in improvement Martin's culture media shaking vase, is shaken under 28 DEG C of cultures
Culture about 7 days is swung,
The step that spreads cultivation of black-koji mould are as follows: the bacterium solution after activation is crossed on improvement Martin's agar slant, it is quiet at 30 DEG C
Set culture 6 days；
Then aseptically, with transfer needle from agar medium bacterium colony picking strain, be put into test tube or conical flask
In fluid nutrient medium (improvement Martin's culture medium), slight oscillatory transfer needle is scattered in thallus in fluid nutrient medium.Institute of the present invention
Stating improvement Martin's culture medium is improvement Martin's culture medium prescription conventional in the prior art.
The culture medium prescription of pseudomonad are as follows: gelatin freezes: 18g, peptone 8g, K2SO414g, MgCL21.6g, agar 20g,
Cetrimonium bronmide 0.4g, PH 6, specific training method are as follows: pseudomonad is inoculated on the culture medium, at 35 DEG C
50h is cultivated,
Then aseptically, with transfer needle from agar medium bacterium colony picking strain, be put into test tube or conical flask
Fluid nutrient medium, the fluid nutrient medium is consistent with the culture medium prescription of pseudomonad among the above, and slight oscillatory transfer needle makes thallus
It is scattered in fluid nutrient medium.
The culture medium of sticky red rhodotorula are as follows: glucose 15g, yeast extract 3g, malt extract 5g, ammonium sulfate 2g, phosphorus
Acid dihydride potassium is 0.5g, bitter salt 0.28g, water 970ml, PH 6, specific training method are as follows: quiet at 28 DEG C
Set culture 7 days.
Embodiment 1
Oil-polluted soils microorganism remediation reagent of the present invention weighs raw material according to following parts by weight:
Microorganism reagent: black-koji mould 1L, pseudomonad 1L, candida utili bacterium 1.5L, sticky red rhodotorula 1L；
Physics adjusts reagent: partial size is that charcoal 0.8kg, the partial size of 80 mesh are the dolomite dust 1.5kg of 80 mesh；
Auxiliary material reagent: sophorolipid 1kg, urea 0.2kg, glucose 0.5kg.
The preparation method of above-mentioned oil-polluted soils microorganism remediation reagent, includes the following steps:
The charcoal that the dolomite that partial size is 80 mesh and partial size are 80 mesh is uniformly mixed by step (1), is gone out in 130 DEG C of high temperature
Bacterium is cooled to room temperature, and obtains physics and adjusts reagent finished product；
Step (2): above-mentioned black-koji mould, pseudomonad, candida utili bacterium and rhodotorula glutinis bacterium solution are mixed equal
It is even, urea is added and glucose is uniformly mixed, stationary culture 10h obtains microorganism remediation liquid；
Step (3): by step (2) microorganism remediation liquid and step (1) obtain physics adjust reagent finished product and
Sophorolipid is uniformly mixed, and obtains oil-polluted soils microorganism remediation reagent finished product.
Embodiment 2
Oil-polluted soils microorganism remediation reagent of the present invention weighs raw material according to following parts by weight:
Microorganism reagent: black-koji mould 1.5L, pseudomonas aeruginosa 2L, candida utili bacterium 1L, sticky red rhodotorula
0.8L；
Physics adjusts reagent: partial size is that charcoal 1.5kg, the partial size of 60 mesh are the dolomite dust 0.8kg of 100 mesh；
Auxiliary material reagent: sophorolipid 2kg, urea 0.3kg, glucose 0.3kg.
The preparation method of above-mentioned oil-polluted soils microorganism remediation reagent, includes the following steps:
The charcoal that the dolomite that partial size is 60 mesh and partial size are 100 mesh is uniformly mixed by step (1), in 150 DEG C of high temperature
Sterilizing, is cooled to room temperature, and obtains physics and adjusts reagent finished product；
Step (2): above-mentioned black-koji mould, pseudomonad, candida utili bacterium and rhodotorula glutinis bacterium solution are mixed equal
It is even, urea is added and glucose is uniformly mixed, stationary culture 13h obtains microorganism remediation liquid；
Step (3): by step (2) microorganism remediation liquid and step (1) obtain physics adjust reagent finished product and
Sophorolipid is uniformly mixed, and obtains oil-polluted soils microorganism remediation reagent finished product.
Embodiment 3
Oil-polluted soils microorganism remediation reagent of the present invention weighs raw material according to following parts by weight:
Microorganism reagent: black-koji mould 1.2L, pseudomonad 1.5L, candida utili bacterium 1.2L, sticky red rhodotorula
0.9L；
It includes following raw material that physics, which adjusts reagent: partial size is that charcoal 1kg, the partial size of 70 mesh are the white clouds of 90 mesh
Mountain flour 1kg；
Auxiliary material reagent includes the raw material of following parts by weight: sophorolipid 1.5kg, urea 0.25kg, glucose 0.4kg.
The preparation method of above-mentioned oil-polluted soils microorganism remediation reagent, includes the following steps:
The charcoal that the dolomite that partial size is 70 mesh and partial size are 90 mesh is uniformly mixed by step (1), is gone out in 140 DEG C of high temperature
Bacterium is cooled to room temperature, and obtains physics and adjusts reagent finished product；
Step (2): above-mentioned black-koji mould, pseudomonad, candida utili bacterium and rhodotorula glutinis bacterium solution are mixed equal
It is even, urea is added and glucose is uniformly mixed, stationary culture 15h obtains microorganism remediation liquid；
Step (3): by step (2) microorganism remediation liquid and step (1) obtain physics adjust reagent finished product and
Sophorolipid is uniformly mixed, and obtains oil-polluted soils microorganism remediation reagent finished product.
Product function test
1. carrying out functional trial using embodiment 1-3:
1.1 test indexes: the repairing effect of oil-polluted soils: petroleum residual quantity, the soil weight, soil are against corrosion in soil
Property, P in soil H value.
1.2 test methods: according to oil-polluted soils microorganism remediation reagent: land area 1kg: 10m2Ratio,
It is 100m that Shaanxi Province's Yan'an, which is chosen, by three pieces of areas are chosen in the experimental plot of oil pollution2Region, be respectively designated as dirt
Contaminate domain test area A, Polluted area test section B, Polluted area test section C；
Specific recovery test mode: Polluted area is measured respectively in morning 7:00 on June 15th, 2018,8:00,9:00 and is surveyed
It is against corrosion to try area A, Polluted area test section B, the oil pollution residue amount in the C soil of Polluted area test section, the soil weight, soil
Property, P in soil H value, record as soil conditions value before repairing；
Then the soil of three pieces of test sections is subjected to shallow cut to 20cm, by the oil pollution in the embodiment of the present invention 1,2,3
Edaphon repairs reagent finished product and is sprayed at Polluted area test section A, Polluted area test section B, Polluted area test respectively
In the pitting that area's C shallow cut goes out, then the soil that shallow cut goes out be crushed to partial size less than 15mm, then raked the soil level, so that broken
Soil after broken is uniformly mixed with the oil-polluted soils microorganism remediation reagent finished product for being sprayed at soil surface carries out Polluted Soil
Earth reparation.
In the corresponding measurement Polluted area test section A of September in 2018 morning 7:00 on the 15th, 8:00,9:00, contaminated area
Domain test area B, the oil pollution residue amount in the C soil of Polluted area test section, the soil weight, the erosion durability of soil, P in soil H value,
It records as soil conditions value after reparation；
2. measure object and method
The soil of 1.1 couples of Polluted area test section A-C that front and back is repaired using reagent in embodiment 1-3 carries out petroleum residual
The detection of amount.
1.11 using petroleum residual quantity in VIS-7220 type visible spectrophotometer measurement soil.
The soil of 1.2 couples of Polluted area test section A-C that front and back is repaired using reagent in embodiment 1-3 is carried out bulk density, resisted
The detection of corrosion.
1.21 pairs of soil using the Polluted area test section A-C before and after reagent reparation in embodiment 1-3 carries out bulk density and adopts
It is detected with core cutter method；
The soil of 1.22 couples of Polluted area test section A-C that front and back is repaired using reagent in embodiment 1-3 carries out corrosion stability
Detection method: grogs 50 of selection 0.3-0.5cm partial size are placed on the sieve of 0.3cm diameter in encroached water, remember every 30s
The grogs number for recording avalanche, continuously records 10min, calculates index against corrosion: index against corrosion=(total grogs-avalanche grogs)/total grogs
× 100%；
1.23 measure P in soil H value using national standard NY-T 1377-2007 method.
3. test result
The soil of 3.1 couples of Polluted area test section A-C that front and back is repaired using reagent in embodiment 1-3 carries out petroleum residual
Testing result it is as shown in table 1:
Table 1
| Petroleum concentration (g/kg) in soil | Before reparation | After reparation |
| Embodiment 1 is directed to test section A | 16.03 | 0.48 |
| Embodiment 2 is directed to test section B | 14.23 | 0.30 |
| Embodiment 3 is directed to test section C | 15.14 | 0.46 |
The soil of 3.2 couples of Polluted area test section A-C that front and back is repaired using reagent in embodiment 1-3 is carried out bulk density, resisted
Corrosion, the testing result of pH value are as shown in table 2- table 4:
Table 2- repairs the bulk density (g/cm of the soil of the Polluted area test section A-C of front and back using reagent in embodiment 1-333)
Variation
| Before reparation | After reparation | |
| Embodiment 1 | 1.62 | 1.34 |
| Embodiment 2 | 1.52 | 1.12 |
| Embodiment 3 | 1.58 | 1.24 |
Table 3- repairs the corrosion stability (%) of the soil of the Polluted area test section A-C of front and back using reagent in embodiment 1-3
Variation
| Before reparation | After reparation | |
| Embodiment 1 | 64 | 88 |
| Embodiment 2 | 66 | 78 |
| Embodiment 3 | 70 | 86 |
Table 4- repairs the pH value variation of the soil of the Polluted area test section A-C of front and back using reagent in embodiment 1-3
| Before reparation | After reparation | |
| Embodiment 1 | 6.5 | 7 |
| Embodiment 2 | 6.0 | 7.5 |
| Embodiment 3 | 6.3 | 7.2 |
By table 1-4 it is found that use oil-polluted soils microorganism remediation reagent of the present invention can effectively degrade in soil
Petroleum, and the structural and permeability of soil can also be enhanced, and then reduce the soil weight, and improve the acid of soil
Property, also effectively enhance the corrosion stability of soil.
The life characteristic of specified raw material and microorganism according to the present invention in the present invention, it is determined that reasonable weight ratio
And preparation method, and it is preparation method simple process of the present invention, low in cost, and the oil-polluted soils that the present invention obtains are micro-
Biological prosthetic reagent can effectively degrade the petroleum in soil, while effectively improve the soil permeability, against corrosion of soil
The physicochemical properties such as property, acidity.
Each embodiment in this specification is described in a progressive manner, the highlights of each of the examples are with other
The difference of embodiment, the same or similar parts in each embodiment may refer to each other.For device disclosed in embodiment
For, since it is corresponded to the methods disclosed in the examples, so being described relatively simple, related place is said referring to method part
It is bright.
The foregoing description of the disclosed embodiments enables those skilled in the art to implement or use the present invention.
Various modifications to these embodiments will be readily apparent to those skilled in the art, as defined herein
General Principle can be realized in other embodiments without departing from the spirit or scope of the present invention.Therefore, of the invention
It is not intended to be limited to the embodiments shown herein, and is to fit to and the principles and novel features disclosed herein phase one
The widest scope of cause.
Claims (10)
1. a kind of oil-polluted soils microorganism remediation reagent, which is characterized in that including the following raw material: microorganism reagent, physics
Adjust reagent, auxiliary material reagent；
The microorganism reagent parts by volume: physics adjusts reagent parts by weight: auxiliary material reagent weight ratio is 3.8-7L: 1.6-
3kg:1.5-2.8kg；
The microorganism reagent includes the strain of following parts by volume: 10-15 parts of black-koji mould, 10-20 parts of pseudomonad produce protein vacation
Silk 10-15 parts of saccharomycete, 8-10 parts of sticky red rhodotorula；
The living bacteria count of any strain is hundred million/milliliter of 1-2 in the microorganism reagent；
It includes following raw material that the physics, which adjusts reagent: 8-15 parts of charcoal, 8-15 parts of dolomite dust；
The auxiliary material reagent includes the raw material of following parts by weight: 10-20 parts of sophorolipid, 2-3 parts of urea, 3-5 parts of glucose.
2. a kind of oil-polluted soils microorganism remediation reagent according to claim 1, which is characterized in that the vacation unit cell
Bacterium is pseudomonas aeruginosa.
3. a kind of oil-polluted soils microorganism remediation reagent, the dolomite described in any one of -2 according to claim 1
The partial size of powder is 80-100 mesh.
4. a kind of oil-polluted soils microorganism remediation reagent according to any one of claim 1-3, the charcoal
Partial size be 60-80 mesh.
5. a kind of preparation method of oil-polluted soils microorganism remediation reagent, which comprises the steps of:
Step (1): claim according to the weight ratio of any one of the claim 1-3 oil-polluted soils microorganism remediation reagent
The physics is taken to adjust reagent and the auxiliary material reagent；
Step (2): black-koji mould, pseudomonad, candida utili bacterium, sticky red rhodotorula are activated respectively, spread cultivation；
Step (3): the strain liquid of each strain obtained after spreading cultivation is according to any one of the claim 1-3 oil pollution soil
The parts by volume ratio of earth microorganism remediation reagent is uniformly mixed, and obtains microorganism mixed liquor；
Step (4): the microorganism mixed liquor that step (3) are obtained is uniformly mixed, and urea is added and glucose is uniformly mixed, stands
10-15h is cultivated, microorganism remediation liquid is obtained；
Step (5): dolomite and charcoal are uniformly mixed, and through high-temperature sterilization, are cooled to room temperature, obtain physics adjust reagent at
Product；
Step (6): physics described in microorganism remediation liquid described in step (4) and step (5) is adjusted into reagent finished product and auxiliary
Expect that reagent sophorolipid is uniformly mixed, obtains oil-polluted soils microorganism remediation reagent finished product.
6. a kind of preparation method of oil-polluted soils microorganism remediation reagent according to claim 4, which is characterized in that
The activation step of black-koji mould described in the step (1) are as follows: aspergillus niger is connected in improvement Martin's culture media shaking vase, 25~
30 DEG C of cultures lower shaken cultivation about 5-8 days；
The step that spreads cultivation of the black-koji mould are as follows: the bacterium solution after activation is crossed on improvement Martin's agar slant, 25~30
DEG C stationary culture 5-7 days.
7. a kind of preparation method of oil-polluted soils microorganism remediation reagent according to claim 4, which is characterized in that
The culture medium prescription of pseudomonad described in the step (1) are as follows: gelatin freezes: 15-18g/L, peptone 8-9g/L, K2SO412-
14g/L, MgCL21.5-1.6g/L, agar 16-20g/L, cetrimonium bronmide 03-0.4g/L；PH is 6-8；
Specific training method are as follows: pseudomonad is inoculated on the culture medium, in 35-37 DEG C of culture 49-52h.
8. a kind of preparation method of oil-polluted soils microorganism remediation reagent according to claim 4, which is characterized in that
Candida utili bacterium described in the step (1): activation medium glucose 20g/L, peptone 20g/L, yeast powder 10g/
L, the water of surplus；
Minimal medium: glucose 30g/L, (NH4)2SO45g/L, yeast powder 3g/L, MgSO40.4g/L, K2HPO41.0g/L, it is remaining
The water of amount；
Specific training method: being first inoculated on activation medium, at 27-28 DEG C, sways culture 15-20h；
Again on minimal medium, continue stationary culture 24-26h at 30-32 DEG C.
9. a kind of preparation method of oil-polluted soils microorganism remediation reagent according to claim 4, which is characterized in that
The culture medium of sticky red rhodotorula described in the step (1) are as follows: concentration of glucose 15g/L, yeast extract concentration are 2-3g/L, wheat
Bud cream concentration is 3-5g/L, and ammonium sulfate concentrations 2-3g/L, biphosphate potassium concn is 0.5-0.8g/L, and bitter salt is dense
Degree is 0.26-0.28g/L, the water of surplus；PH is 6-8；
Specific training method are as follows: stationary culture 6-9 days at 28-32 DEG C.
10. a kind of preparation method of oil-polluted soils microorganism remediation reagent according to claim 4, feature exist
In high-temperature sterilization temperature described in the step (4) is 130-150 DEG C.
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