CN102757921A - Bacillus cereus and application thereof - Google Patents
Bacillus cereus and application thereof Download PDFInfo
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- CN102757921A CN102757921A CN2012102643783A CN201210264378A CN102757921A CN 102757921 A CN102757921 A CN 102757921A CN 2012102643783 A CN2012102643783 A CN 2012102643783A CN 201210264378 A CN201210264378 A CN 201210264378A CN 102757921 A CN102757921 A CN 102757921A
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Abstract
The invention discloses a bacillus cereus Bacillus cereus 4033. The conservation number of the bacillus cereus is CGMCCNO.6074. The invention also discloses the application of the bacillus cereus in degrading oil pollutant. The bacillus cereus is derived from soil, so that the strain rapidly becomes dominant strain after entering into the soil, occupies a certain ecological niche, effectively promotes degradation of oil hydrocarbon, optimizes the soil microbe system, improves the physical behavior of soil, enhances the biological activity of soil, and restores hardened and degraded soil. The bacillus cereus is a dominant bacteria which is selected by using crude oil as the sole carbon source, the property of crude oil can be changed in the process of growth and multiplication, components of crude oil can be changed, the functions of emulsifying, wetting and dispersing crude oil are realized, and the oil pollutant in the soil can be degraded in a short time.
Description
Technical field
The invention belongs to technical field of bioengineering, be specifically related to a kind of bacillus cereus and application thereof.
Background technology
The mikrobe recovery technique of oil-polluted soils; Obtained large development in recent ten years abroad; But the petroleum hydrocarbon class pollutant degradation speed is slow; Cause governance process longer perdurability, be an outstanding difficult point of this technology always, therefore seeks the research focus that the highly effective petroleum degradation bacteria is current oil-polluted soils mikrobe recovery technique.
Summary of the invention
Technical problem to be solved by this invention is the deficiency to above-mentioned prior art, and a kind of bacillus cereus that can be used for the degraded oil pollutent is provided.This bacterial strain is to be the dominant strain that sole carbon source screens with crude oil, and bacterial strain can change the character of crude oil in the growth and breeding process, and the component of crude oil is changed, and plays emulsification, wetting and disperse the effect of crude oil.
For solving the problems of the technologies described above; The technical scheme that the present invention adopts is: a kind of bacillus cereus; It is characterized in that said bacillus cereus is Bacillus cereus 4033, deposit number CGMCCNo.6074; Preservation day is on May 3rd, 2012, and depositary institution is China Committee for Culture Collection of Microorganisms common micro-organisms center.
1, this bacterial strain has following character:
Morphological specificity: bacterium colony is white in color smooth, moistening round bacterium, diameter 1mm～2mm; Cell is shaft-like, cell dia 1.0 μ m～1.5 μ m.
Physiological and biochemical property: gram positive bacterium.
2, the screening method of this bacterial strain is:
Step 1, enrichment: will be from the Xi'an Weiyang District Chang An University Wei Shui school district Shaanxi Province's underground water and the testing ground soil petroleum pollution 10000mg/kg of ecotope ERC repair the soil sample of gathering in red clover rhizosphere reparation district, the 10cm left and right sides, district and take by weighing 10.0g with sterile manner; Insert in 100.0mL oil/inorganic salt liquid substratum, place 28 ± 1 ℃, 130r/min constant temperature shaking table enrichment culture 7d; Draw the 5mL nutrient solution then and transfer again in fresh oil/inorganic salt liquid substratum, carry out 3 enrichments, domestication again, each 7d; Consisting of of said oil/inorganic salt liquid substratum: NaNO
30.15%, (NH
4) SO
40.15%, K
2HPO
40.1%, MgSO
47H
2O 0.05%, and KCl 0.05%, FeSO
47H
2O 0.001%, CaCl
20.0002%, zero(ppm) water 1000mL, crude oil (prolong oil dehydrating crude oil) 0.5% more than is mass percent, medium pH value 7.0～7.5, the 105KPa 20min～25min that sterilizes;
Step 2, screening and purifying: the enrichment culture liquid after the enrichment culture in the step 1 is pressed the decimal dilution method dilution, get 10
-4, 10
-5, 10
-63 each 0.1mL of dilution nutrient solution coat on oil/inorganic salt solid plate substratum; Place and cultivate 48h in 37 ± 1 ℃ of constant incubators, choose the single bacterium colony of advantage of different colours and form, the separation of on oil/inorganic salt solid plate substratum, repeatedly ruling; Obtain the consistent purifying bacterial strain of form with purifying; The purifying bacterial strain is stored in the enterprising row of beef extract-peptone solid medium further screens, activation, obtains bacterial strain, for use; Consisting of of said oil/inorganic salt solid plate substratum: NaNO
30.15%, (NH
4) SO
40.15%, K
2HPO
40.1%, MgSO
47H
2O 0.05%, and KCl 0.05%, FeSO
47H
2O 0.001%, CaCl
20.0002%, agar 2%, zero(ppm) water 1000mL, crude oil (prolong oil dehydrating crude oil) 0.5% more than is mass percent, medium pH value 7.0～7.5, the 105KPa 20min～25min that sterilizes;
Step 3, multiple sieve: the bacterial strain that activation in the step 2 is good is inoculated in 100mL respectively and contains in the inorganic salt liquid substratum of 1wt% n-dodecane and 1wt% hexanaphthene; 28 ± 1 ℃, the cultivation of 130r/min shaking table; After the obvious muddiness of nutrient solution; Getting the lmL nutrient solution is inoculated in the inorganic salt solid medium that contains 1wt% n-dodecane and 1wt% hexanaphthene and cultivates; And then the inoculation of cultivating cultivated in 100mL contains the inorganic salt liquid substratum of 1wt% n-dodecane and 1wt% hexanaphthene, so repeat 3 times, preserve the bacterial strain that finally still can make the inorganic salt liquid substratum change muddiness that contains n-dodecane and hexanaphthene.
3, the authentication method of this bacterial strain is:
Step 1, extract total DNA:
101, get incubated overnight bacterial strain (at the most 2 * 10
9Individual), the centrifugal 1min of 10000r/min collects thalline, add 180 μ L bacteriolyze solution (the 20mg/mL N,O-Diacetylmuramidase, 20mMTris-HCl pH8,2.5mMEDTA, 1%TritonX-100), resuspended bacterium liquid, 37 ℃ of water-bath 30min～60min; In resuspended bacterium liquid, add 20 μ L Proteinase K solution (10mg/mL) then, the mixing that fully vibrates, 56 ℃ of water-bath 30min are to the complete cracking of cell; Then add 200 μ L solution B D, the mixing that fully vibrates, 70 ℃ of water-bath 10min; Add 200 μ L absolute ethyl alcohols again, mixing fully vibrates;
102, adsorption column is put into collection tube, all add in the adsorption column with the bacterium liquid of pipettor after with vibration mixing in 101, leave standstill 2min, the centrifugal 3min of 12000r/min outwells filtrating; In adsorption column, add 500 μ L solution PW, the centrifugal 1min of 10000r/min outwells filtrating; In adsorption column, add 500 μ L rinsing solution Wash buffer, the centrifugal 1min of 10000r/min outwells filtrating; Adsorption column is put into collection tube,, remove residual rinsing liquid Washbuffer in the centrifugal 2min of 12000r/min;
103, take out adsorption column, put into the centrifuge tube of a new 1.5mL, add the elutriant Elution buffer of 50 μ L preheatings (60 ℃), leave standstill 3min, the centrifugal 1min of 10000r/min room temperature collects dna solution; Adopting concentration is that 2% agarose detects the total DNA that extracts, and clip size is about 13kb;
The pcr amplification of step 2,16SrDNA: with the total DNA that extracts in the step 1 is that template is carried out pcr amplification, the PCR reaction system: dna profiling 10pmol, upstream primer (10 μ M) 1 μ l, downstream primer (10 μ M) 1 μ l; DNTP (10Mm) 1 μ l; 10 * Taq reaction Buffer, 5 μ l; Taq (5U/ μ l) 0.25 μ l; Add water to 50 μ l; PCR reaction conditions: preparatory 98 ℃ of 5min of sex change; The 95 ℃ of 35s that circulate, 55 ℃ of 35s, 72 ℃ of 30s, 35 circulations, last 72 ℃ are extended 8min, 4 ℃ of preservations; The primer sequence of said pcr amplification is:
Upstream primer (SEQ ID NO.1): 5'-CAGAGTTTGATCCTGGCT-3';
Downstream primer (SEQ ID NO.2): 5'-AGGAGGTGATCCAGCCGCA-3';
Step 3, bacterial strain 16SrDNA sequential analysis and comparison: the product (about clip size 1.4kb) to pcr amplification in the step 2 check order (assist accomplishing) by Sangon Biotech (Shanghai) Co., Ltd.; The 16SrDNA sequence of the bacillus cereus of having delivered among 16SrDNA portion gene sequence that obtains (SEQ ID NO.3) and the GenBank (Bacillus cereus) is carried out homology relatively, and similarity reaches 100%.
Comprehensive Physiology and biochemistry is identified, 16SrDNA sequential analysis qualification result, identifies that bacterial strain of the present invention is a bacillus cereus.
The present invention also provides the above-mentioned application that is used for bacillus cereus at the degraded oil pollutent.
The present invention compared with prior art has the following advantages:
1, bacillus cereus of the present invention comes from soil; Therefore this bacterial strain becomes dominant microflora after getting into soil very soon, occupies certain niche, effectively promotes the degraded of petroleum hydrocarbon; Optimize the soil microorganisms system; Improve the physical behavior of soil, strengthen the biological activity of soil, repair the soil that hardens and degenerate.
2, bacillus cereus of the present invention is to be the dominant strain that sole carbon source screens with crude oil, and bacterial strain can change the character of crude oil in the growth and breeding process, and the component of crude oil is changed, and plays emulsification, wetting and disperse the effect of crude oil.
3, the bacillus cereus of the present invention petroleum pollution in the soil of can degrading at short notice.
Through embodiment, technical scheme of the present invention is done further detailed description below.
Embodiment
The screening of bacterial strain:
Step 1, enrichment: will be from the Xi'an Weiyang District Chang An University Wei Shui school district Shaanxi Province's underground water and the testing ground soil petroleum pollution 10000mg/kg of ecotope ERC repair the soil sample of gathering in red clover rhizosphere reparation district, the 10cm left and right sides, district and take by weighing 10.0g with sterile manner; Insert in 100.0mL oil/inorganic salt liquid substratum, place 28 ± 1 ℃, 130r/min constant temperature shaking table enrichment culture 7d; Draw the 5mL nutrient solution then and transfer again in fresh oil/inorganic salt liquid substratum, carry out 3 enrichments, domestication again, each 7d; Consisting of of said oil/inorganic salt liquid substratum: NaNO
30.15%, (NH
4) SO
40.15%, K
2HPO
40.1%, MgSO
47H
2O 0.05%, and KCl 0.05%, FeSO
47H
2O 0.001%, CaCl
20.0002%, zero(ppm) water 1000mL, crude oil (prolong oil dehydrating crude oil) 0.5% more than is mass percent, medium pH value 7.0～7.5, the 105KPa 20min～25min that sterilizes;
Step 2, screening and purifying: the enrichment culture liquid after the enrichment culture in the step 1 is pressed the decimal dilution method dilution, get 10
-4, 10
-5, 10
-63 each 0.1mL of dilution nutrient solution coat on oil/inorganic salt solid plate substratum; Place and cultivate 48h in 37 ± 1 ℃ of constant incubators, choose the single bacterium colony of advantage of different colours and form, the separation of on oil/inorganic salt solid plate substratum, repeatedly ruling; Obtain the consistent purifying bacterial strain of form with purifying; The purifying bacterial strain is stored in the enterprising row of beef extract-peptone solid medium further screens, activation, obtains bacterial strain, for use; Consisting of of said oil/inorganic salt solid plate substratum: NaNO
30.15%, (NH
4) SO
40.15%, K
2HPO
40.1%, MgSO
47H
2O 0.05%, and KCl 0.05%, FeSO
47H
2O 0.001%, CaCl
20.0002%, agar 2%, zero(ppm) water 1000mL, crude oil (prolong oil dehydrating crude oil) 0.5% more than is mass percent, medium pH value 7.0～7.5, the 105KPa 20min～25min that sterilizes;
Step 3, multiple sieve: the bacterial strain that activation in the step 2 is good is inoculated in 100mL respectively and contains in the inorganic salt liquid substratum of 1wt% n-dodecane and 1wt% hexanaphthene; 28 ± 1, the 130r/min shaking table is cultivated; After the obvious muddiness of nutrient solution; Get the lmL nutrient solution be inoculated into contain the 1wt% n-dodecane and with the inorganic salt solid medium of 1wt% hexanaphthene in cultivate; And then the inoculation of cultivating cultivated in 100mL contains the inorganic salt liquid substratum of 1wt% n-dodecane and 1wt% hexanaphthene, so repeat 3 times, preserve the bacterial strain that finally still can make the inorganic salt liquid substratum change muddiness that contains n-dodecane and hexanaphthene.
The evaluation of bacterial strain:
Step 1, extract total DNA:
101, get incubated overnight bacterial strain (at the most 2 * 10
9Individual), the centrifugal 1min of 10000r/min collects thalline, add 180 μ L bacteriolyze solution (the 20mg/mL N,O-Diacetylmuramidase, 20mMTris-HCl pH8,2.5mMEDTA, 1%TritonX-100), resuspended bacterium liquid, 37 ℃ of water-bath 30min～60min; In resuspended bacterium liquid, add 20 μ L Proteinase K solution (10mg/mL) then, the mixing that fully vibrates, 56 ℃ of water-bath 30min are to the complete cracking of cell; Then add 200 μ L solution B D, the mixing that fully vibrates, 70 ℃ of water-bath 10min; Add 200 μ L absolute ethyl alcohols again, mixing fully vibrates;
102, adsorption column is put into collection tube, all add in the adsorption column with the bacterium liquid of pipettor after with vibration mixing in 101, leave standstill 2min, the centrifugal 3min of 12000r/min outwells filtrating; In adsorption column, add 500 μ L solution PW, the centrifugal 1min of 10000r/min outwells filtrating; In adsorption column, add 500 μ L rinsing solution Wash buffer, the centrifugal 1min of 10000r/min outwells filtrating; Adsorption column is put into collection tube, in the centrifugal 2min of 12000r/min, the rinsing liquid Washbuffer that leaves away residual;
103, take out adsorption column, put into the centrifuge tube of a new 1.5mL, add the elutriant Elution buffer of 50 μ L preheatings (60 ℃), leave standstill 3min, the centrifugal 1min of 10000r/min room temperature collects dna solution; Adopting concentration is that 2% agarose detects the total DNA that extracts, and clip size is about 13kb;
The pcr amplification of step 2,16SrDNA: with the total DNA that extracts in the step 1 is that template is carried out pcr amplification, the PCR reaction system: dna profiling 10pmol, upstream primer (10 μ M) 1 μ l, downstream primer (10 μ M) 1 μ l; DNTP (10Mm) 1 μ l; 10 * Taq reaction Buffer, 5 μ l; Taq (5U/ μ l) 0.25 μ l; Add water to 50 μ l; PCR reaction conditions: preparatory 98 ℃ of 5min of sex change; The 95 ℃ of 35s that circulate, 55 ℃ of 35s, 72 ℃ of 30s, 35 circulations, last 72 ℃ are extended 8min, 4 ℃ of preservations; The primer sequence of said pcr amplification is:
Upstream primer (SEQ ID NO.1): 5'-CAGAGTTTGATCCTGGCT-3';
Downstream primer (SEQ ID NO.2): 5'-AGGAGGTGATCCAGCCGCA-3';
Step 3, bacterial strain 16SrDNA sequential analysis and comparison: the product (about clip size 1.4kb) to pcr amplification in the step 2 check order (assist accomplishing) by Sangon Biotech (Shanghai) Co., Ltd.; The 16SrDNA sequence of the bacillus cereus of having delivered among 16SrDNA portion gene sequence that obtains (SEQ ID NO.3) and the GenBank (Bacillus cereus) is carried out homology relatively, and similarity reaches 100%.
Comprehensive Physiology and biochemistry is identified, 16SrDNA sequential analysis qualification result, identifies that bacterial strain of the present invention is a bacillus cereus, and called after bacillus cereus Bacillus cereus 4033, deposit number CGMCC No.6074.
The effect test of bacillus cereus of the present invention:
1, bacillus cereus is tested the degradation capability of crude oil:
Bacillus cereus of the present invention is inoculated in the inorganic salt liquid substratum that contains crude oil, in 28 ± 1 ℃, 130rpm fermentation culture 7 days, substratum is formed: NaNO
30.15%, (NH
4) SO
40.15%, K
2HPO
40.1%, MgSO
47H
2O 0.05%, and KCl 0.05%, FeSO
47H
2O 0.001%, CaCl
20.0002%, crude oil 1%, zero(ppm) water 1000mL more than is mass percent, and pH 7.0～7.5, sterilization 20min～25min; Compare with the substratum of not inoculating bacillus cereus, the color of fermented liquid is obviously deepened, and the crude oil in the fermented liquid of inoculation bacillus cereus has the advantages that not hang bottle.This explanation, bacillus cereus of the present invention are the sole carbon source growth with crude oil, have changed the character of crude oil, make the component of crude oil that variation take place.Crude oil after the inoculation bacillus cereus fermentation is examined under a microscope, found to have in the crude oil bacterium liquid to exist, bacterium exists on the water-oil interface, and is carbon source with oil, conforms to produce oil substances, thereby plays emulsification, wetting and disperse the effect of crude oil.
2, bacillus cereus is tested the degraded of substratum PetroChina Company Limited.:
Bacillus cereus of the present invention is inoculated in the inorganic salt liquid substratum that contains crude oil, in 28 ± 1 ℃, 130rpm fermentation culture 7 days, the inorganic salt liquid substratum that contains crude oil is formed: NaNO
30.15%, (NH
4) SO
40.15%, K
2HPO
40.1%, MgSO
47H
2O 0.05%, KCl0.05%g, FeSO
47H
2O 0.001%, CaCl
20.0002%, crude oil 1%, zero(ppm) water 1000mL more than is mass percent, and pH 7.0～7.5, sterilization 20min～25min; Getting the 10mL fermented liquid then respectively is inoculated in the fresh inorganic salt liquid substratum that contains crude oil of 100mL (substratum form the same), contains the inorganic salt liquid substratum of n-dodecane and contain in the inorganic salt liquid substratum of hexanaphthene; In 28 ± 1 ℃; 130rpm fermentation culture 7 days, the inorganic salt liquid substratum that contains n-dodecane is formed: NaNO
30.15%, (NH
4) SO
40.15%, K
2HPO
40.1%, MgSO
47H
2O0.05%, KCl 0.05%g, FeSO
47H
2O 0.001%, CaCl
20.0002%, n-dodecane 1%, zero(ppm) water 1000mL more than is mass percent, and pH 7.0～7.5, sterilization 20min～25min; The inorganic salt liquid substratum that contains hexanaphthene is formed: NaNO
30.15%, (NH
4) SO
40.15%, K
2HPO
40.1%, MgSO
47H
2O 0.05%, KCl 0.05%g, FeSO
47H
2O 0.001%, CaCl
20.0002%, hexanaphthene 1%, zero(ppm) water 1000mL more than is mass percent, pH7.0～7.5, sterilization 20min～25min; With nonvaccinated three kinds of liquid nutrient mediums is blank; In 28 ± 1 ℃; The 130rpm fermentation culture was taken a sample in 7 days, detected the fermented liquid of inoculation bacillus cereus and total petroleum hydrocarbon (TPH) degradation rate, straight-chain paraffin degradation rate and the naphthenic hydrocarbon degradation rate of blank respectively, and the result sees table 1.
The oil degradation rate and the function thereof of table 1 bacterial strain
| The TPH degradation rate | The straight-chain paraffin degradation rate | The naphthenic hydrocarbon degradation rate | |
| 4033 bacterial strains | 38.5% | 43.8% | 14.7% |
| Blank | 9.8% | 12.2% | 10.7% |
From table 1, can find out; The degradation rate of total petroleum hydrocarbon has improved 28.7% than blank in the presence of bacillus cereus of the present invention; The degradation rate of straight-chain paraffin and naphthenic hydrocarbon all has a more substantial increase, and bacillus cereus of the present invention can effectively degrade straight-chain paraffin and naphthenic hydrocarbon are described.
3, bacillus cereus is to the degraded situation of soil PetroChina Company Limited.
Bacillus cereus of the present invention is inoculated in 37 ± 1 ℃ of activation 36h in the beef extract-peptone solid medium; Bacterial classification after the activation is added in the different dustiness soil according to 5% (mass percent) ratio; All add 5% wheat bran in the soil; The ratio of C in the soil: N: P is 100: 5: 1, and N, P source are respectively NH
4NO
3And K
2HPO
4, the dustiness of soil PetroChina Company Limited. is respectively 0.3%, 0.7% and 1.5%, and keeping soil moisture content is 20%～25%, is blank with the soil that does not add bacterial classification.Through 21 days degraded, the degradation rate result of TPH (total petroleum hydrocarbon) was as shown in table 2.
The oil degradation rate of table 2 soil petroleum pollution bacterial strain
| The petroleum pollution degree | 0.3% | 0.7% | 1.5% |
| 4033 bacterial strains | 39.8% | 40.2% | 38.1% |
| Blank | 15.6% | 12.2% | 10.7% |
Degraded through bacillus cereus; TPH under 3 kinds of petroleum pollution degree conditions in the soil all has tangible reduction; Improved 24.2%, 28.0% and 27.4% than blank respectively, can find out that bacillus cereus can reach degradation effect preferably at soil a middle or short term.
In sum; Bacillus cereus of the present invention comes from soil, so becomes dominant microflora very soon behind this bacterial strain entering soil, occupies certain niche; Can degrade at short notice petroleum pollution in the soil; Effectively promote the degraded of petroleum hydrocarbon, optimize the soil microorganisms system, improve the physical behavior of soil.
The above; It only is preferred embodiment of the present invention; Be not that the present invention is done any restriction, every according to inventing technical spirit to any simple modification, change and equivalent structure variation that above embodiment did, all still belong in the protection domain of technical scheme of the present invention.
Claims (2)
1. a bacillus cereus is characterized in that, said bacillus cereus is Bacillus cereus 4033, deposit number CGMCC No.6074.
2. the application of bacillus cereus in the degraded oil pollutent according to claim 1.
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| CN 201210264378 CN102757921B (en) | 2012-07-27 | 2012-07-27 | Bacillus cereus and application thereof |
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| CN102757921B CN102757921B (en) | 2013-10-16 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103834588A (en) * | 2014-01-07 | 2014-06-04 | 南京农业大学 | Bacillus cereus stain with polycyclic aromatic hydrocarbon degradation capability and application thereof |
| CN110468082A (en) * | 2019-09-19 | 2019-11-19 | 四川农业大学 | A kind of A Shi bacillus OCB-6 and its application |
| CN114657099A (en) * | 2022-04-01 | 2022-06-24 | 福州大学 | Petroleum hydrocarbon degrading strain and screening and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1374399A (en) * | 2002-04-11 | 2002-10-16 | 中国科学院武汉病毒研究所 | Waxy bacillus and its prepn |
| CN1417325A (en) * | 2002-12-14 | 2003-05-14 | 山西省农业科学院植物保护研究所 | Waxy bacillus strain and microbial bactericide producing process with the strain |
| CN1580241A (en) * | 2004-05-17 | 2005-02-16 | 大庆油田有限责任公司 | Bacterium for degrading petroleum and it use |
-
2012
- 2012-07-27 CN CN 201210264378 patent/CN102757921B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1374399A (en) * | 2002-04-11 | 2002-10-16 | 中国科学院武汉病毒研究所 | Waxy bacillus and its prepn |
| CN1417325A (en) * | 2002-12-14 | 2003-05-14 | 山西省农业科学院植物保护研究所 | Waxy bacillus strain and microbial bactericide producing process with the strain |
| CN1580241A (en) * | 2004-05-17 | 2005-02-16 | 大庆油田有限责任公司 | Bacterium for degrading petroleum and it use |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103834588A (en) * | 2014-01-07 | 2014-06-04 | 南京农业大学 | Bacillus cereus stain with polycyclic aromatic hydrocarbon degradation capability and application thereof |
| CN103834588B (en) * | 2014-01-07 | 2016-04-13 | 南京农业大学 | One strain has bacillus cereus and the application thereof of degrading polycyclic aromatic hydrocarbons ability |
| CN110468082A (en) * | 2019-09-19 | 2019-11-19 | 四川农业大学 | A kind of A Shi bacillus OCB-6 and its application |
| CN114657099A (en) * | 2022-04-01 | 2022-06-24 | 福州大学 | Petroleum hydrocarbon degrading strain and screening and application thereof |
| CN114657099B (en) * | 2022-04-01 | 2023-02-21 | 福州大学 | Petroleum hydrocarbon degrading strain and screening and application thereof |
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| Publication number | Publication date |
|---|---|
| CN102757921B (en) | 2013-10-16 |
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