CN106957809A - The biological synergetic restorative procedure of soil organic contamination - Google Patents
The biological synergetic restorative procedure of soil organic contamination Download PDFInfo
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- CN106957809A CN106957809A CN201710340828.5A CN201710340828A CN106957809A CN 106957809 A CN106957809 A CN 106957809A CN 201710340828 A CN201710340828 A CN 201710340828A CN 106957809 A CN106957809 A CN 106957809A
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- bacterial agent
- microbial bacterial
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- bacillus pumilus
- gordonia
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- 239000002689 soil Substances 0.000 title claims abstract description 28
- 238000000034 method Methods 0.000 title claims abstract description 17
- 238000011109 contamination Methods 0.000 title claims abstract description 15
- 230000002195 synergetic effect Effects 0.000 title claims abstract description 7
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 83
- 230000001580 bacterial effect Effects 0.000 claims abstract description 80
- 230000000813 microbial effect Effects 0.000 claims abstract description 72
- 241000194103 Bacillus pumilus Species 0.000 claims abstract description 51
- 241000203749 Gordonia bronchialis Species 0.000 claims abstract description 37
- 150000001335 aliphatic alkanes Chemical class 0.000 claims abstract description 19
- 235000013305 food Nutrition 0.000 claims abstract description 19
- 241000196324 Embryophyta Species 0.000 claims abstract description 17
- 241000589636 Xanthomonas campestris Species 0.000 claims abstract description 16
- 241001656677 Gordonia alkanivorans Species 0.000 claims abstract description 11
- 241000028625 Gordonia amicalis Species 0.000 claims abstract description 11
- 241000982689 Pseudoxanthomonas japonensis Species 0.000 claims abstract description 11
- 241000894006 Bacteria Species 0.000 claims description 19
- 239000002609 medium Substances 0.000 claims description 17
- 239000007788 liquid Substances 0.000 claims description 16
- 230000000694 effects Effects 0.000 claims description 13
- RTZKZFJDLAIYFH-UHFFFAOYSA-N ether Substances CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 11
- 239000001963 growth medium Substances 0.000 claims description 11
- 230000008635 plant growth Effects 0.000 claims description 11
- 239000004094 surface-active agent Substances 0.000 claims description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 238000004321 preservation Methods 0.000 claims description 4
- 238000012549 training Methods 0.000 claims description 4
- BWDBEAQIHAEVLV-UHFFFAOYSA-N 6-methylheptan-1-ol Chemical compound CC(C)CCCCCO BWDBEAQIHAEVLV-UHFFFAOYSA-N 0.000 claims description 3
- 229910019142 PO4 Inorganic materials 0.000 claims description 3
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 claims description 3
- 238000012258 culturing Methods 0.000 claims description 3
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 3
- 239000010452 phosphate Substances 0.000 claims description 3
- 238000012216 screening Methods 0.000 claims description 3
- 229910052938 sodium sulfate Inorganic materials 0.000 claims description 3
- 235000011152 sodium sulphate Nutrition 0.000 claims description 3
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims description 2
- 229910052698 phosphorus Inorganic materials 0.000 claims description 2
- 239000011574 phosphorus Substances 0.000 claims description 2
- 239000011734 sodium Substances 0.000 claims description 2
- 229910052708 sodium Inorganic materials 0.000 claims description 2
- BDHFUVZGWQCTTF-UHFFFAOYSA-M sulfonate Chemical compound [O-]S(=O)=O BDHFUVZGWQCTTF-UHFFFAOYSA-M 0.000 claims description 2
- DCXXMTOCNZCJGO-UHFFFAOYSA-N tristearoylglycerol Chemical compound CCCCCCCCCCCCCCCCCC(=O)OCC(OC(=O)CCCCCCCCCCCCCCCCC)COC(=O)CCCCCCCCCCCCCCCCC DCXXMTOCNZCJGO-UHFFFAOYSA-N 0.000 claims 2
- 150000004702 methyl esters Chemical class 0.000 claims 1
- 239000003209 petroleum derivative Substances 0.000 abstract description 19
- 244000005700 microbiome Species 0.000 abstract description 17
- 230000012010 growth Effects 0.000 abstract description 9
- 239000002068 microbial inoculum Substances 0.000 abstract description 9
- 238000000354 decomposition reaction Methods 0.000 abstract description 8
- 230000035784 germination Effects 0.000 abstract description 6
- 208000035143 Bacterial infection Diseases 0.000 abstract description 4
- 102000018997 Growth Hormone Human genes 0.000 abstract description 4
- 108010051696 Growth Hormone Proteins 0.000 abstract description 4
- 230000009286 beneficial effect Effects 0.000 abstract description 4
- 239000000122 growth hormone Substances 0.000 abstract description 4
- 239000003921 oil Substances 0.000 description 32
- 235000015097 nutrients Nutrition 0.000 description 23
- 230000000052 comparative effect Effects 0.000 description 10
- 230000015556 catabolic process Effects 0.000 description 9
- 238000006731 degradation reaction Methods 0.000 description 9
- 239000002131 composite material Substances 0.000 description 8
- 238000011081 inoculation Methods 0.000 description 8
- 239000004215 Carbon black (E152) Substances 0.000 description 6
- AFCARXCZXQIEQB-UHFFFAOYSA-N N-[3-oxo-3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propyl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C(CCNC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)N1CC2=C(CC1)NN=N2 AFCARXCZXQIEQB-UHFFFAOYSA-N 0.000 description 6
- 229930195733 hydrocarbon Natural products 0.000 description 6
- 150000002430 hydrocarbons Chemical class 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 239000000417 fungicide Substances 0.000 description 5
- 230000000855 fungicidal effect Effects 0.000 description 4
- 239000013049 sediment Substances 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 241000193830 Bacillus <bacterium> Species 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 241000219823 Medicago Species 0.000 description 2
- 235000017587 Medicago sativa ssp. sativa Nutrition 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 239000010779 crude oil Substances 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 239000004575 stone Substances 0.000 description 2
- 241000283725 Bos Species 0.000 description 1
- 241000726221 Gemma Species 0.000 description 1
- 241000635201 Pumilus Species 0.000 description 1
- 235000021355 Stearic acid Nutrition 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 241000219793 Trifolium Species 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 239000003344 environmental pollutant Substances 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 1
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 231100000719 pollutant Toxicity 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000005067 remediation Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000002269 spontaneous effect Effects 0.000 description 1
- 239000008117 stearic acid Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 239000002351 wastewater Substances 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
- B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
- B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
- B09C1/105—Reclamation of contaminated soil microbiologically, biologically or by using enzymes using fungi or plants
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
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- Environmental & Geological Engineering (AREA)
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- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Soil Sciences (AREA)
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Abstract
A kind of biological synergetic restorative procedure of soil organic contamination the invention discloses microbial bacterial agent and using the microbial inoculum, the microbial bacterial agent contains the active component of effective dose, the active component includes the bacillus pumilus (Bacillus pumilus) that deposit number is CGMCC NO.13208, preserving number is CGMCC No.4794 friendly Gordonia bronchialis (Gordonia amicalis), the false Xanthomonas campestris (Pseudoxanthomonas japonensis) of Japan that the food alkane Gordonia bronchialis (Gordonia alkanivorans) and preserving number that preserving number is CGMCC No.4796 are CGMCC No.4797；The method that the disclosure additionally provides the application of the microbial bacterial agent and its administers oil pollution, the microbial bacterial agent can secrete the germination and growth that growth hormone promotes plant, while can suppress the generation of bacterial diseases of plants using mentioned microorganism microbial inoculum；The microbial bacterial agent and plant can be mutually promoted growth for administering oil-polluted soils simultaneously, efficient-decomposition petroleum hydrocarbon, improvement oil pollution is more beneficial for.
Description
Technical field
This disclosure relates to organic contamination technical field of biological remediation, in particular it relates to a kind of microbial bacterial agent and its be used for
Administer the antimicrobial plant cooperative repair method of soil organic contamination.
Background technology
The world today, oil has become one of topmost energy of the mankind, and substantial amounts of oil and its processed goods enter to bury
Earth causes the oil pollution of soil, and harm is brought to the mankind or even whole biosphere, so that the environmental problem that becomes international.
It is biological prosthetic to be just increasingly subject to pay attention to as a kind of potential clean technologies efficiently, inexpensive in the research of oil pollution.
Biological prosthetic (Bioremediation) refers to by biological (particularly microorganism) catalytic degradation organic pollution, so as to repair
The process of the controlled or spontaneous progress of contaminated environment or the pollutant in elimination environment.
China was from 1975, successively to Jiaozhou Bay of Qingdao, the Bohai Sea, PORT OF XIAMEN, the Huanghai Sea and East Sea oil degradation microorganism
Quantity, distribution, species composition and influence degradation factors etc. investigated.In degraded of the influence microorganism to petroleum hydrocarbon
Principal element in, the physiological property of microorganism is to determine one of key factor of decomposing petroleum hydrocarbon ability.Separate at present
Obtained microbial strains are also relatively low to the adaptability of environmental condition, and petroleum hydrocarbon degradation in the environment is inefficient, therefore,
In the urgent need to developing a kind of biological synergetic reparation for the microbial bacterial agent and soil organic contamination for being capable of efficient degradation petroleum hydrocarbon
Method.
The content of the invention
The purpose of the disclosure is to provide a kind of strong microbial bacterial agent of degradation capability to petroleum hydrocarbon and the organic dirt of soil
The biological synergetic restorative procedure of dye, it can effectively handle oil pollution.
Present disclose provides a kind of microbial bacterial agent, the microbial bacterial agent contains the active component of effective dose, the work
Property composition comprising deposit number be CGMCC NO.13208 bacillus pumilus (Bacillus pumilus), preserving number be
CGMCC No.4794 friendly Gordonia bronchialis (Gordonia amicalis), the food alkane that preserving number is CGMCC No.4796
The false Xanthomonas campestris of Japan that Gordonia bronchialis (Gordonia alkanivorans) and preserving number are CGMCC No.4797
(Pseudoxanthomonas japonensis)。
The disclosure additionally provides application of the microbial bacterial agent as described above in organic contamination is administered.
The disclosure additionally provides application of the microbial bacterial agent as described above in plant growth is promoted.
The disclosure additionally provides a kind of method for administering oil pollution, soil or water of this method including the use of oil pollution
Body planting plants, are administered simultaneously microbial bacterial agent as described above.
The disclosure additionally provides a kind of bacillus pumilus, the guarantor of the bacillus pumilus (Bacillus pumilus)
It is CGMCC NO.13208 to hide numbering.
By above-mentioned technical proposal, between the microbial bacterial agent that the disclosure is provided and the association between microorganism and plant
Same-action contributes to efficient degradation petroleum hydrocarbon organic carbon macromolecular, and the microbial bacterial agent can secrete growth hormone promotion
The germination and growth of plant, while the generation of bacterial diseases of plants can be suppressed using mentioned microorganism microbial inoculum；Will be described micro-
Bacteria agent and plant can mutually promote growth applied to administering oil pollution simultaneously, be more beneficial for efficient-decomposition petroleum hydrocarbon,
Administer oil pollution.
Other feature and advantage of the disclosure will be described in detail in subsequent embodiment part.
Biomaterial preservation
Bacillus pumilus (Bacillus pumilus) is the inventor of the disclosure from oil-polluted soils in the disclosure
In isolated pure culture, its deposit number is CGMCC NO.13208, and preservation date is on October 27th, 2016, preservation
Unit is China Committee for Culture Collection of Microorganisms's common micro-organisms center, and address is BeiChen West Road, Chaoyang District, BeiJing City 1
Number institute 3, Classification And Nomenclature is bacillus pumilus (Bacillus pumilus).Wherein, preserving number is CGMCC No.4794's
Friendly Gordonia bronchialis (Gordonia amicalis), the food alkane Gordonia bronchialis that preserving number is CGMCC No.4796
The false Xanthomonas campestris of Japan that (Gordonia alkanivorans) and preserving number are CGMCC No.4797
The biological deposits information of (Pseudoxanthomonas japonensis) is in Publication No. CN102250770A patent
Disclosed in document.
Embodiment
Embodiment of this disclosure is described in detail below.It should be appreciated that described herein specific
Embodiment is merely to illustrate and explained the disclosure, is not limited to the disclosure.
Disclosure first aspect provides a kind of microbial bacterial agent, it is characterised in that the microbial bacterial agent contains effectively
The active component of amount, the active component includes the bacillus pumilus (Bacillus that deposit number is CGMCC NO.13208
Pumilus friendly Gordonia bronchialis (Gordonia amicalis) that), preserving number is CGMCC No.4794, preserving number are CGMCC
No.4796 food alkane Gordonia bronchialis (Gordonia alkanivorans) and preserving number is false for CGMCC No.4797 Japan
Xanthomonas campestris (Pseudoxanthomonas japonensis).
By above-mentioned technical proposal, the synergy between the microbial bacterial agent that the disclosure is provided contributes to efficient degradation stone
Petroleum hydrocarbon class organic carbon macromolecular, and the microbial bacterial agent can secrete the germination and growth that growth hormone promotes plant, simultaneously
The generation of bacterial diseases of plants can be suppressed using mentioned microorganism microbial inoculum；The microbial bacterial agent and plant are applied simultaneously
It can be mutually promoted growth in administering oil pollution, be more beneficial for efficient-decomposition petroleum hydrocarbon, improvement oil pollution.
According to the disclosure in a first aspect, in order that each bacterial strain can act synergistically, not to other in the microbial bacterial agent
The growth of bacterial strain produces suppression, the bacillus pumilus, friendly Gordonia bronchialis, the false yellow unit cell of food alkane Gordonia bronchialis and Japan
The quantity ratio of bacterium is 1：(0.15-7.5)：(0.05-5)：(0.05-5).
According to the disclosure in a first aspect, in order that the microbial bacterial agent reaches micro- life described in the purpose of fast onset effect
Number of viable in thing microbial inoculum needs to reach certain level, and the liquid of solid microbial or 1mL relative to 1g is micro-
Bacteria agent, total viable count is (0.1-2.5) × 1010It is individual；Preferably (0.5-1) × 1010It is individual.
According to the disclosure in a first aspect, the microbial bacterial agent also contains the culture medium for being used for keeping microbial activity；Institute
It can be various conventional solid mediums or fluid nutrient medium to state culture medium；Preferably, the culture medium is beef extract egg
At least one in white peptone culture medium, oil screening and culturing medium and PDA culture medium.For example, the microbial bacterial agent can be preserved
In solid beef-protein medium；The oil screening and culturing medium is suitable for the microbial bacteria of oil pollution at this to screen
Agent；PDA culture medium can be used during growth in order to activate, accelerate the microbial bacterial agent；While in order to keep micro- life
Thing microbial inoculum is applied to administer oil pollution initial stage with higher vigor, can use liquid beef-protein medium.
According to the disclosure in a first aspect, in order to accelerate the speed that the microbial bacterial agent decomposes petroleum hydrocarbon, the microorganism
Microbial inoculum also contains surfactant；On the basis of the gross weight of the microbial bacterial agent, the content of the surfactant is
0.1-0.5 weight %.
According to the disclosure in a first aspect, under preferable case, the surfactant is fatty alcohol-ether sodium sulfate, stearic acid first
At least one in ester APEO sodium sulfonate, Di-phosphorus pentonide and isooctanol ether phosphate.
Disclosure second aspect provides application of the microbial bacterial agent as described above in organic contamination is administered, described micro-
Bacteria agent is strong due to petroleum hydrocarbon capacity of decomposition, can be applied individually to any in improvement oil pollution.
The disclosure third aspect provides application of the described microbial bacterial agent in plant growth is promoted, the microorganism
Promotion plant growth can be used alone in microbial inoculum, its can secrete growth hormone IAA GA etc. promote the germination and growth of plant,
The generation of bacterial diseases of plants can be suppressed using mentioned microorganism microbial inoculum simultaneously.
The disclosure third aspect provide a kind of biological synergetic restorative procedure of soil organic contamination, this method including the use of
The soil or water body planting plants of organic contamination, are administered simultaneously microbial bacterial agent as described above；Preferably, the organic contamination
For oil pollution.The microbial bacterial agent and plant can be mutually promoted growth applied to administering oil pollution simultaneously, more had
Beneficial to efficient-decomposition petroleum hydrocarbon, oil pollution is administered.
Disclosure fourth aspect provides a kind of bacillus pumilus, the bacillus pumilus (Bacillus pumilus)
Deposit number be CGMCC NO.13208；The bacillus pumilus is screened by the disclosed invention people from oil-polluted soils
Obtain, the bacterial strain has very strong degradation capability to petroleum hydrocarbon.
The present invention is further described by the following examples.
Embodiment 1
Counted using the gross weight of the strain as 100%, by the short and small gemma that the deposit number is CGMCC NO.13208
Bacillus (Bacillus pumilus) is inoculated in 100mL liquid beef-protein medium；By the training after the inoculation
Support based on cultivated under conditions of 28 DEG C, 180 revs/min the concentration of bacillus pumilus in culture medium described in 1 angel for 2.5 ×
1010Individual/mL；Counted using the gross weight of the microbial bacterial agent as 100%, 0.1 weight % fat is added into the culture medium
Ether sulfate prepares the bacillus pumilus agent of the present embodiment.
Embodiment 2
Bacillus pumilus (Bacillus pumilus), preserving number by the deposit number for CGMCC NO.13208
For CGMCC No.4794 friendly Gordonia bronchialis (Gordonia amicalis), the food alkane that preserving number is CGMCC No.4796
The false Xanthomonas campestris of Japan that hydrocarbon Gordonia bronchialis (Gordonia alkanivorans) and preserving number are CGMCC No.4797
(Pseudoxanthomonas japonensis) is inoculated in 100mL liquid beef-protein medium respectively；By institute
State and cultivate 1 angel its bacterial concentration under conditions of the culture after inoculation is based on 28 DEG C, 180 revs/min and reach 1 × 109Individual/mL,
By the nutrient solution containing friendly Gordonia bronchialis described in the nutrient solution containing bacillus pumilus described in 9 volume %, 1.5 volume %,
Contain the false Xanthomonas campestris of Japan described in nutrient solution and 44.75 volume % containing food alkane Gordonia bronchialis described in 44.75 volume %
Nutrient solution well mixed be configured to 100mL composite bacteria agents；Total viable count in microbial bacterial agent described in 1mL is 0.1 × 1010
It is individual.
Embodiment 3
Bacillus pumilus (Bacillus pumilus), preserving number by the deposit number for CGMCC NO.13208
For CGMCC No.4794 friendly Gordonia bronchialis (Gordonia amicalis), the food alkane that preserving number is CGMCC No.4796
The false Xanthomonas campestris of Japan that hydrocarbon Gordonia bronchialis (Gordonia alkanivorans) and preserving number are CGMCC No.4797
(Pseudoxanthomonas japonensis) is inoculated in 100mL liquid beef-protein medium respectively；By institute
State and cultivate 1 angel its bacterial concentration under conditions of the culture after inoculation is based on 28 DEG C, 180 revs/min and reach 5 × 1010Individual/mL,
By the nutrient solution containing friendly Gordonia bronchialis described in the nutrient solution containing bacillus pumilus described in 5.8 volume %, 44 volume %,
Culture containing the false Xanthomonas campestris of Japan described in nutrient solution, 0.1 volume % containing food alkane Gordonia bronchialis described in 0.1 volume %
Liquid and the liquid beef-protein medium of surplus, which are well mixed, is configured to 100mL composite bacteria agents；Microbial bacterial agent described in 1mL
In total viable count be 2.5 × 1010It is individual.
Embodiment 4
Bacillus pumilus (Bacillus pumilus), preserving number by the deposit number for CGMCC NO.13208
For CGMCC No.4794 friendly Gordonia bronchialis (Gordonia amicalis), the food alkane that preserving number is CGMCC No.4796
The false Xanthomonas campestris of Japan that hydrocarbon Gordonia bronchialis (Gordonia alkanivorans) and preserving number are CGMCC No.4797
(Pseudoxanthomonas japonensis) is inoculated in 100mL liquid beef-protein medium respectively；By institute
State and cultivate 1 angel its bacterial concentration under conditions of the culture after inoculation is based on 28 DEG C, 180 revs/min and reach 1 × 1010Individual/mL,
By the training containing friendly Gordonia bronchialis described in the nutrient solution containing bacillus pumilus described in 6.25 volume %, 18.75 volume %
Contain the false yellow unit cell of Japan described in nutrient solution, 12.5 volume % containing food alkane Gordonia bronchialis described in nutrient solution, 12.5 volume %
The nutrient solution of bacterium and the liquid beef-protein medium of surplus, which are well mixed, is configured to 100mL composite bacteria agents；It is micro- described in 1mL
Total viable count in bacteria agent is 0.5 × 1010It is individual.
Embodiment 5
Bacillus pumilus (Bacillus pumilus), preserving number by the deposit number for CGMCC NO.13208
For CGMCC No.4794 friendly Gordonia bronchialis (Gordonia amicalis), the food alkane that preserving number is CGMCC No.4796
The false Xanthomonas campestris of Japan that hydrocarbon Gordonia bronchialis (Gordonia alkanivorans) and preserving number are CGMCC No.4797
(Pseudoxanthomonas japonensis) is inoculated in 100mL liquid beef-protein medium respectively；By institute
State and cultivate 1 angel its bacterial concentration under conditions of the culture after inoculation is based on 28 DEG C, 180 revs/min and reach 1 × 1010Individual/mL,
By the nutrient solution containing friendly Gordonia bronchialis described in the nutrient solution containing bacillus pumilus described in 25 volume %, 25 volume %,
Culture containing the false Xanthomonas campestris of Japan described in nutrient solution and 25 volume % containing food alkane Gordonia bronchialis described in 25 volume %
Liquid is well mixed to be configured to 100mL composite bacteria agents；Total viable count in microbial bacterial agent described in 1mL is 1 × 1010It is individual.
Embodiment 6
Bacillus pumilus (Bacillus pumilus), preserving number by the deposit number for CGMCC NO.13208
For CGMCC No.4794 friendly Gordonia bronchialis (Gordonia amicalis), the food alkane that preserving number is CGMCC No.4796
The false Xanthomonas campestris of Japan that hydrocarbon Gordonia bronchialis (Gordonia alkanivorans) and preserving number are CGMCC No.4797
(Pseudoxanthomonas japonensis) is inoculated in 100mL liquid beef-protein medium respectively；By institute
State and cultivate 1 angel its bacterial concentration under conditions of the culture after inoculation is based on 28 DEG C, 180 revs/min and reach 1010Individual/mL, by 25
Nutrient solution containing bacillus pumilus, the nutrient solution containing friendly Gordonia bronchialis described in 25 volume %, 25 bodies described in volume %
Nutrient solution containing the false Xanthomonas campestris of Japan described in nutrient solution and 25 volume % containing food alkane Gordonia bronchialis described in product % is mixed
Conjunction is uniformly configured to 100mL composite bacteria agents；Counted using the gross weight of the microbial bacterial agent as 100%, into the composite bacteria agent
The isooctanol ether phosphate for adding 0.5 weight % prepares the microbial bacterial agent of the present embodiment；In microbial bacterial agent described in 1mL
Total viable count be 1 × 1010It is individual.
Embodiment 7
Bacillus pumilus (Bacillus pumilus), preserving number by the deposit number for CGMCC NO.13208
For CGMCC No.4794 friendly Gordonia bronchialis (Gordonia amicalis), the food alkane that preserving number is CGMCC No.4796
The false Xanthomonas campestris of Japan that hydrocarbon Gordonia bronchialis (Gordonia alkanivorans) and preserving number are CGMCC No.4797
(Pseudoxanthomonas japonensis) is inoculated in 100mL liquid beef-protein medium respectively；By institute
State and cultivate 1 angel its bacterial concentration under conditions of the culture after inoculation is based on 28 DEG C, 180 revs/min and reach 1010Individual/more than mL,
By the training containing friendly Gordonia bronchialis described in the nutrient solution containing bacillus pumilus described in 6.25 volume %, 18.75 volume %
Contain the false yellow unit cell of Japan described in nutrient solution, 12.5 volume % containing food alkane Gordonia bronchialis described in nutrient solution, 12.5 volume %
The nutrient solution of bacterium and the liquid beef-protein medium of surplus, which are well mixed, is configured to 100mL composite bacteria agents；With described micro-
The gross weight of bacteria agent is 100% to count and 0.1 weight % fatty alcohol-ether sodium sulfate is added into the composite bacteria agent be prepared into
To the microbial bacterial agent of the present embodiment；Total viable count in microbial bacterial agent described in 1mL is 0.5 × 1010It is individual.
Comparative example 1
Method according to embodiment 1 prepares microbial bacterial agent, unlike, the deposit number is CGMCC NO.13208
Bacillus pumilus replaces with the bacillus pumilus for the numbering PDBP-2 that inventor voluntarily separates.
Comparative example 2
Method according to embodiment 3 prepares microbial bacterial agent, unlike, the deposit number is CGMCC NO.13208
Bacillus pumilus replaces with the bacillus pumilus for the numbering PDBP-3 that inventor voluntarily separates.
Testing example 1
The microbial bacterial agent that this testing example is used to determine in embodiment 1-7 and comparative example 1-2 decomposes the effect of petroleum hydrocarbon
Rate.
Oil-polluted soils sample, every part of each 10g (oil content 1.25g) are weighed, the pedotheque is to pick up from carat
Agate is according to the ground crude oil contaminated soil around the producing well in oil field, oil sediment and the long-term topsoil soil for stacking oil sediment according to 1：1：
The 1 mixed oil-polluted soils sample of ratio；The oil-polluted soils sample is separately added into equipped with 100mL liquid oxen
In the 250mL triangular flasks of meat extract peptone, while being inoculated with respectively micro- in the embodiment 1-7, comparative example 1 or comparative example 2
Biological each 3 bottles；Cultivate 5 days, detected in the culture medium under the conditions of the culture is based on into 28 DEG C, 180 revs/min after inoculation
Oil content (g), concrete outcome is shown in Table 1.
Table 1
Compared through embodiment 1-7 in table 1 with comparative example 1-2 as can be seen that the preserving number that contains that the disclosure is provided is CGMCC
The microbial bacterial agent of No.13208 bacillus pumilus has the effect for preferably decomposing petroleum hydrocarbon；Contain in embodiment 2-5
The complex micro organism fungicide of a variety of bacterial strains has higher petroleum hydrocarbon decomposition efficiency compared with Example 1；It is highly preferred that embodiment
Viable count described in 4-7 in complex micro organism fungicide is (0.5-1.0) × 1010Petroleum hydrocarbon decomposition efficiency is higher during individual/mL；Most
Preferably, there is more preferable stone when 0.1-0.5 weight % surfactant is added with microbial bacterial agent described in embodiment 6-7
Petroleum hydrocarbon discomposing effect.
Testing example 2
The microbial bacterial agent that this testing example is used to determine in embodiment 1-7 and comparative example 1-2 promotes plant growth
Efficiency.
Take every part of 100 alfalfa seeds to be put into the 100mm of 3 filter paper of place mat culture dish, contain after being diluted with sterilized water
The microbial bacterial agent processing for 10 volume % is measured, using sterilized water as control, keeps filter paper to infiltrate and cultivate 3 days at room temperature
After determine potentiality of seed, after 5 days determine germination percentage and root it is long, concrete outcome is shown in Table 2.
Table 2
Compared through embodiment 1-7 in table 2 with comparative example 1-2 as can be seen that the preserving number that contains that the disclosure is provided is CGMCC
The microbial bacterial agent of No.13208 bacillus pumilus has the effect for preferably promoting plant growth；Contain in embodiment 2-7
Having the complex micro organism fungicides of a variety of bacterial strains, effect more preferably, under preferable case, is combined described in embodiment 4-7 compared with Example 1
Viable count in microbial bacterial agent is (0.5-1.0) × 1010Promote the effect of plant growth more preferable during individual/mL.
Testing example 3
This testing example is the microbial bacterial agent and plant combined processing petroleum wastewater in embodiment 1-7 and comparative example 1-2
Contaminate the effect of soil.
Oil-polluted soils sample, every part of each 100g (oil content 5g) are weighed, the pedotheque is to pick up from a carat agate
According to the ground crude oil contaminated soil around the producing well in oil field, oil sediment and the long-term topsoil soil for stacking oil sediment according to 1：1：1
The mixed oil-polluted soils sample of ratio；The oil-polluted soils sample is layered in 100mm culture dish, taken
100 alfalfa seeds are respectively put into the culture dish of place mat petroleum-contaminated soil, are handled with 10 volume % microbial bacterial agent pouring, with nothing
Bacterium water is used as control；Potentiality of seed is determined after keeping oil-polluted soils to be infiltrated and cultivated 5 days at room temperature, after 10 days
Determine the resolution ratio of germination percentage, root length and the oil of clover.Concrete outcome is shown in Table 3.
Table 3
Compared through embodiment 1-7 in table 3 with comparative example 1-2 as can be seen that the preserving number that contains that the disclosure is provided is CGMCC
The microbial bacterial agent of No.13208 bacillus pumilus and plant combined improvement oil pollution effect are more preferable, while being conducive to planting
The growth of thing；The complex micro organism fungicide containing a variety of bacterial strains promotes plant growth, divided compared with Example 1 in embodiment 2-7
The effect for solving greasy dirt is more preferable；It is highly preferred that viable count in complex micro organism fungicide described in embodiment 4-7 for (0.5-1.0) ×
1010Promote the effect of plant growth, decomposition greasy dirt more preferable during individual/mL；Most preferably, in microbial bacterial agent described in embodiment 6-7
There is best oil discomposing effect during surfactant added with 0.1-0.5 weight %.
The preferred embodiment of the disclosure described in detail above, still, the disclosure are not limited in above-mentioned embodiment
Detail, in the range of the technology design of the disclosure, a variety of simple variants can be carried out with technical scheme of this disclosure, this
A little simple variants belong to the protection domain of the disclosure.
It is further to note that each particular technique feature described in above-mentioned embodiment, in not lance
In the case of shield, it can be combined by any suitable means.In order to avoid unnecessary repetition, the disclosure to it is various can
The combination of energy no longer separately illustrates.
In addition, can also be combined between a variety of embodiments of the disclosure, as long as it is without prejudice to originally
Disclosed thought, it should equally be considered as disclosure disclosure of that.
Claims (10)
1. a kind of microbial bacterial agent, it is characterised in that the microbial bacterial agent contains the active component of effective dose, the activity into
Subpackage is CGMCC containing the bacillus pumilus (Bacillus pumilus), preserving number that deposit number is CGMCC NO.13208
No.4794 friendly Gordonia bronchialis (Gordonia amicalis), the food alkane Ge Dengshi that preserving number is CGMCC No.4796
The false Xanthomonas campestris of Japan that bacterium (Gordonia alkanivorans) and preserving number are CGMCC No.4797
(Pseudoxanthomonas japonensis)。
2. microbial bacterial agent according to claim 1, wherein, the bacillus pumilus, friendly Gordonia bronchialis, food alkane
The quantity ratio of the false Xanthomonas campestris of Gordonia bronchialis and Japan is 1：(0.15-7.5)：(0.05-5)：(0.05-5).
3. the microbial bacterial agent according to claim 1 or 2, wherein, the liquid of microbial bacterial agent or 1mL relative to 1g
Body microbial bacterial agent, total viable count is (0.1-2.5) × 1010It is individual；Preferably (0.5-1) × 1010It is individual.
4. the microbial bacterial agent according to claim 1 or 2, wherein, the microbial bacterial agent, which also contains, to be used to keep micro-
The culture medium of bioactivity；Preferably, the culture medium is beef-protein medium, oil screening and culturing medium and PDA trainings
Support at least one in base.
5. microbial bacterial agent according to claim 1 or 2, wherein, the microbial bacterial agent also contains surfactant；With
On the basis of the gross weight of the microbial bacterial agent, the content of the surfactant is 0.1-0.5 weight %.
6. microbial bacterial agent according to claim 5, wherein, the surfactant is fatty alcohol-ether sodium sulfate, tristearin
At least one in sour methyl esters sodium sulfonate of ethoxylated, Di-phosphorus pentonide and isooctanol ether phosphate.
7. application of the microbial bacterial agent in organic contamination is administered in claim 1-6 described in any one.
8. application of the microbial bacterial agent in plant growth is promoted in claim 1-6 described in any one.
9. a kind of biological synergetic restorative procedure of soil organic contamination, it is characterised in that this method is including the use of organic contamination
Soil or water body planting plants, are administered simultaneously the microbial bacterial agent described in any one in claim 1-6；Preferably, it is described
Organic contamination is oil pollution.
10. a kind of bacillus pumilus, it is characterised in that the preservation of the bacillus pumilus (Bacillus pumilus) is compiled
Number be CGMCC NO.13208.
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| CN112159060A (en) * | 2020-09-29 | 2021-01-01 | 重庆大学 | Microorganism-combined plant-type composite deodorant for black-odor rivers and preparation method thereof |
| CN112400546A (en) * | 2021-01-22 | 2021-02-26 | 中国农业科学院农业环境与可持续发展研究所 | Ecological rod and application thereof |
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