CN108795807B - A strain of Lysobacter SCSIO17111 that produces exopolysaccharide and can fix sand and its application - Google Patents
A strain of Lysobacter SCSIO17111 that produces exopolysaccharide and can fix sand and its application Download PDFInfo
- Publication number
- CN108795807B CN108795807B CN201810609500.3A CN201810609500A CN108795807B CN 108795807 B CN108795807 B CN 108795807B CN 201810609500 A CN201810609500 A CN 201810609500A CN 108795807 B CN108795807 B CN 108795807B
- Authority
- CN
- China
- Prior art keywords
- sand
- scsio
- lysobacter
- molten bacillus
- sand fixation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 239000004576 sand Substances 0.000 title claims abstract description 54
- 241000863031 Lysobacter Species 0.000 title abstract description 7
- 229920002444 Exopolysaccharide Polymers 0.000 title 1
- 239000002689 soil Substances 0.000 claims abstract description 24
- 206010039509 Scab Diseases 0.000 claims abstract description 9
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 39
- 241000894006 Bacteria Species 0.000 claims description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 14
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 12
- 241001248650 Lysobacter sp. Species 0.000 claims description 11
- 239000001963 growth medium Substances 0.000 claims description 11
- 239000011780 sodium chloride Substances 0.000 claims description 6
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 5
- 239000008103 glucose Substances 0.000 claims description 5
- 239000002904 solvent Substances 0.000 claims description 4
- 239000012137 tryptone Substances 0.000 claims description 4
- AIUDWMLXCFRVDR-UHFFFAOYSA-N dimethyl 2-(3-ethyl-3-methylpentyl)propanedioate Chemical compound CCC(C)(CC)CCC(C(=O)OC)C(=O)OC AIUDWMLXCFRVDR-UHFFFAOYSA-N 0.000 claims description 3
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 3
- 150000004676 glycans Chemical class 0.000 abstract description 18
- 229920001282 polysaccharide Polymers 0.000 abstract description 18
- 239000005017 polysaccharide Substances 0.000 abstract description 18
- 244000005700 microbiome Species 0.000 abstract description 15
- 230000000694 effects Effects 0.000 abstract description 14
- 238000000034 method Methods 0.000 abstract description 7
- 239000000126 substance Substances 0.000 abstract description 6
- 230000000813 microbial effect Effects 0.000 abstract description 3
- 239000002245 particle Substances 0.000 abstract description 2
- 238000010276 construction Methods 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 14
- 244000132059 Carica parviflora Species 0.000 description 12
- 235000014653 Carica parviflora Nutrition 0.000 description 12
- 230000001580 bacterial effect Effects 0.000 description 9
- 239000000523 sample Substances 0.000 description 9
- 241000196324 Embryophyta Species 0.000 description 8
- 241000192700 Cyanobacteria Species 0.000 description 6
- 230000015572 biosynthetic process Effects 0.000 description 6
- 239000008367 deionised water Substances 0.000 description 6
- 229910021641 deionized water Inorganic materials 0.000 description 6
- 238000011160 research Methods 0.000 description 6
- 241001086727 Lysobacter firmicutimachus Species 0.000 description 5
- 230000002776 aggregation Effects 0.000 description 5
- 238000004220 aggregation Methods 0.000 description 5
- 238000002835 absorbance Methods 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 239000000306 component Substances 0.000 description 3
- 238000002955 isolation Methods 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 230000001954 sterilising effect Effects 0.000 description 3
- 108020004465 16S ribosomal RNA Proteins 0.000 description 2
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 241000192656 Nostoc Species 0.000 description 2
- 241000192497 Oscillatoria Species 0.000 description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
- 241000192608 Phormidium Species 0.000 description 2
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 2
- 241000589636 Xanthomonas campestris Species 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- -1 are retained Substances 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 230000006866 deterioration Effects 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000001704 evaporation Methods 0.000 description 2
- 230000008020 evaporation Effects 0.000 description 2
- 239000012467 final product Substances 0.000 description 2
- 239000002054 inoculum Substances 0.000 description 2
- 229910052698 phosphorus Inorganic materials 0.000 description 2
- 239000011574 phosphorus Substances 0.000 description 2
- 239000011148 porous material Substances 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 230000002265 prevention Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000013049 sediment Substances 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 239000001117 sulphuric acid Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 241000186046 Actinomyces Species 0.000 description 1
- 201000004384 Alopecia Diseases 0.000 description 1
- 241000099686 Azotobacter sp. Species 0.000 description 1
- 241001037822 Bacillus bacterium Species 0.000 description 1
- 241000565779 Cylindrospermum Species 0.000 description 1
- 241000192128 Gammaproteobacteria Species 0.000 description 1
- 240000008917 Glycyrrhiza uralensis Species 0.000 description 1
- 235000000554 Glycyrrhiza uralensis Nutrition 0.000 description 1
- 101710094902 Legumin Proteins 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 235000008331 Pinus X rigitaeda Nutrition 0.000 description 1
- 235000011613 Pinus brutia Nutrition 0.000 description 1
- 241000018646 Pinus brutia Species 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- 241000192142 Proteobacteria Species 0.000 description 1
- 241000719329 Trentepohlia Species 0.000 description 1
- 235000009754 Vitis X bourquina Nutrition 0.000 description 1
- 235000012333 Vitis X labruscana Nutrition 0.000 description 1
- 240000006365 Vitis vinifera Species 0.000 description 1
- 235000014787 Vitis vinifera Nutrition 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 241001148470 aerobic bacillus Species 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000013065 commercial product Substances 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 229960000935 dehydrated alcohol Drugs 0.000 description 1
- NEKNNCABDXGBEN-UHFFFAOYSA-L disodium;4-(4-chloro-2-methylphenoxy)butanoate;4-(2,4-dichlorophenoxy)butanoate Chemical compound [Na+].[Na+].CC1=CC(Cl)=CC=C1OCCCC([O-])=O.[O-]C(=O)CCCOC1=CC=C(Cl)C=C1Cl NEKNNCABDXGBEN-UHFFFAOYSA-L 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000012533 medium component Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000002068 microbial inoculum Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000003020 moisturizing effect Effects 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 238000010899 nucleation Methods 0.000 description 1
- 230000010355 oscillation Effects 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 229920000573 polyethylene Polymers 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 239000003516 soil conditioner Substances 0.000 description 1
- 238000004162 soil erosion Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- 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
-
- 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
-
- 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
-
- E—FIXED CONSTRUCTIONS
- E02—HYDRAULIC ENGINEERING; FOUNDATIONS; SOIL SHIFTING
- E02D—FOUNDATIONS; EXCAVATIONS; EMBANKMENTS; UNDERGROUND OR UNDERWATER STRUCTURES
- E02D3/00—Improving or preserving soil or rock, e.g. preserving permafrost soil
Landscapes
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Medicinal Chemistry (AREA)
- Structural Engineering (AREA)
- Agronomy & Crop Science (AREA)
- Environmental & Geological Engineering (AREA)
- Soil Sciences (AREA)
- General Life Sciences & Earth Sciences (AREA)
- Mining & Mineral Resources (AREA)
- Paleontology (AREA)
- Civil Engineering (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明公开了一种产胞外多糖可固沙的溶杆菌SCSIO 17111及其应用。本发明的溶杆菌SCSIO 17111于2017年8月18日保藏于中国微生物菌种保藏管理委员会普通微生物中心(CGMCC)，地址：北京市朝阳区北辰西路1号院3号，中国科学院微生物研究所，保藏编号：CGMCC NO.14537。本发明所提供的溶杆菌SCSIO 17111可以使沙粒团聚并保持相对稳定的状态，从而起到固沙的效果，可以应用于防治干旱、半干旱地区沙漠化和岛礁生物土壤结皮的构建。相较于物理固沙、化学固沙和植被的培植，利用微生物结皮固沙作为新型固沙方式，具有适应性强、成本低、见效快等优势。
The invention discloses a lysobacter SCSIO 17111 that produces extracellular polysaccharide and can fix sand and its application. The lysobacteria SCSIO 17111 of the present invention was deposited on August 18, 2017 in the General Microbiology Center (CGMCC) of the China Microorganism Culture Collection Management Committee, address: No. 3, No. 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences , deposit number: CGMCC NO.14537. The lysobacteria SCSIO 17111 provided by the present invention can make the sand particles agglomerate and maintain a relatively stable state, thereby achieving the effect of sand fixation, and can be applied to prevent and control desertification in arid and semi-arid areas and the construction of biological soil crusts on islands and reefs. Compared with physical sand fixation, chemical sand fixation and vegetation cultivation, the use of microbial crust sand fixation as a new sand fixation method has the advantages of strong adaptability, low cost and quick effect.
Description
Technical field:
The invention belongs to microorganism fields, and in particular to one plant of extracellular polysaccharide has the molten bacillus SCSIO for ability of fixing the sand
17111 and its application.
Background technique:
Molten Bacillus (Lysobacter) belongs to Proteobacteria, γ-deformation Gammaproteobacteria, Xanthomonas campestris mesh, Xanthomonas campestris section.It is molten
Bacillus genus strain cell is in thin rod shape, and size is generally 0.2~0.5 × 1.0~15 (some can reach 70) μm, atrichia, tool
Coasting ability, it is flexible, it is Gram-negative, aerobic bacteria, G+C content is high in genomic DNA, reach 61.7%~
70.7%.Molten Bacillus bacterium colony has compared with strong tack, and color is generally cream-colored, pink or yellowish-brown.
Over nearly two, 30 years, as the size of population increases, city size constantly expands, the aggravation of economical activities of mankind, right
The soil of Semi-humid areas arid, semiarid and with Droughts occurs for the irrational utilization of land resource and destruction
It degenerates, i.e. " desertification " is also in land deterioration.In many environmental problems of the mankind, desertification be disaster the most serious it
One, global prone soil just with annual 5~70,000 square kilometres of speed expanded, there is 1,000,000,000 or more people, 40% or more land
Surface is influenced by desertification.
Arid, the expansion of semiarid zone desertification are increasingly severe, lead to land deterioration, soil fertility, two packing spaces
Property reduce, extreme environment and socio-economic factor exacerbate the degeneration in soil.The main method of preventing and fixing sand is that physics is solid at present
Husky, chemical sand-fixing and vegetation cultivation, but the cost is relatively high, and Arid&semi-arid area precipitation is insufficient, evaporation is strong, plants trees
The survival rate of afforestation is lower.In such a way that microorganism skinning fixes the sand as novel fix the sand, it is adaptable it is strong, at low cost, take effect
The advantages such as fast.Microorganism in the skinning of desert is both the important component of the participant that skinning is formed and skinning, has and increases
Strong soil aggregation stability improves surface soil water situation, prevents the effects of soil erosion.Existing research shows soil knot
Why skin can be formed, and the polysaccharose substance with microorganism and secreted by it has inseparable relationship.Bacterium is in soil
At the initial stage that skinning is formed, important function is played, a large amount of extracellular polymeric i.e. exocellular polysaccharide can be secreted in its metabolic process.
The cementing soil particle of polysaccharide material energy makes aggregate, and bonding is played in the formation and stabilization process of aggregate and is made
With.The research that related bacterium is fixed the sand is it has been reported that Pan etc. will be from Gurbantunggut Desert, xinjiang, china Biological Soil Crusts bottom point
The one plant of Oligotrophic bacteria separated out is made microbial inoculum and is sprayed on desert surface, can play and fix the sand and slow down soil water evaporation
Effect, to play certain improvement result to desert Environment.Zhang Xuemei etc. will separate pure from ancient capital Xi'an
Change the obtained azotobacteria (Azotobacter sp.) of one plant of extracellular polysaccharide and bacterium solution is made is sprayed at drift sand surface and forms
One layer of microorganism skinning has water conservation, the effect of keeping a full stand of seedings to the perennial arid desert plant such as epheday intermedia, Glycyrrhiza Uralensis.Wu etc. will
After from the microbionation isolated in the biological breadcrust of desert to field Shatian, the poly- of heterotrophic microorganism in topsoil have stimulated
Collection, bacterium, the quantity of actinomyces and total phosphorus, available nitrogen, available phosphorus contents dramatically increase, and have to soil and fix the sand, are retained, chemical defence
The effect corroded is learned, while being played the role of to the recovery of biological breadcrust effective.
At this stage, the microorganism for having been used for fixing the sand is concentrated mainly on Cyanophyta, mainly includes filamentous cyanobacteria, such as Nostoc
(Nostoc), Oscillatoria (Oscillatoria), column spore Trentepohlia (Cylindrospermum) and Phormidium (Phormidium)
Deng.Cyanobacteria has been had studied decades as the use of soil conditioner by spells, as people gradually recognize cyanobacteria
Importance in biological breadcrust forming process becomes the emphasis of concern again recently.Cyanobacteria is inoculated into sandy land surface, it can
Aggregate is formed by adhesive attraction of the algal filament to organic matters such as the Mechanical entanglements of the grains of sand and cell polysaccharide, is made of aggregate
Biological breadcrust can play sand fixation.Scientific research personnel gropes to research and develop on outer drift sand out of office the technical method for being inoculated with cyanobacteria, achieves
Certain achievement, the relevant technologies are applied to repair desertification habitat.Though molten bacillus can produce exocellular polysaccharide, have no at present to molten
The correlative study of the ability of fixing the sand of bacillus is reported.
Summary of the invention:
It separates and obtains from Chinese Hainan Province's Sansha City Nansha Islands Biological Soil Crusts the object of the present invention is to provide one plant
Extracellular polysaccharide and 17111 bacterial strain of molten bacillus (Lysobacter sp.) SCSIO that can fix the sand, to fix the sand and biological
Soil crust provides new microbial resources.
It is micro- that molten bacillus (Lysobacter sp.) SCSIO 17111 of the invention was preserved in China on August 18th, 2017
Biological inoculum preservation administration committee common micro-organisms center (CGMCC), address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Number, Institute of Microorganism, Academia Sinica, deposit number: CGMCC No.14537.
The present invention is according to phend-sulphuric acid (Phenol-SulphuricAcidMethod), using glucose standard curve,
17111 yield of extracellular polysaccharide of bacterial strain SCSIO is measured, its yield of extracellular polysaccharide is 0.08 ± 0.01mg/ml as the result is shown.With molten
The TSB culture solution of bacillus SCSIO 17111 is uniformly sprayed at partial size less than 1.25mm and is greater than the sterile coral sand of 0.20mm
When, by culture in 55 days, it is found that it can effectively promote the formation of skinning, the skinning of formation with a thickness of 6.00 ±
1.32mm；When culture solution is equably sprayed on sterile coral sand of the partial size less than 0.20mm, by culture in 55 days, formed
Skinning with a thickness of 2.33 ± 0.58mm.Although the skinning thickness of the latter is not so good as the former, the soil aggregation object of the latter is stablized
Property it is stronger, i.e., to soil aggregation object carry out dry screen when, the aggregate of the latter is able to maintain preferable stability, and the former is more fragile
It is broken.
Therefore, a second object of the present invention is to provide molten bacillus SCSIO 17111 to fix the sand and/or establish biological soil
Application in skinning.
It is preferred that application method is the bacterium solution for first cultivating molten bacillus SCSIO 17111, then bacterium solution is inoculated into what needs fixed the sand
Region.
Further preferably, the bacterium solution of the molten bacillus SCSIO 17111 of culture is with the molten bacillus of TSB culture medium culture
SCSIO 17111 obtains bacterium solution, the TSB culture medium are as follows: tryptone 17.0g/L, soy peptone 3.0g/L, grape
Sugared 2.5g/L, sodium chloride 5.0g/L, K2HPO42.5g/L, solvent are water, pH7.3 ± 0.2.
Molten bacillus SCSIO 17111 provided by the present invention can make the grains of sand reunite and maintain a relatively stable state, from
And the effect fixed the sand is played, it can be applied to the building of prevention and treatment arid, semiarid zone desertification and islands and reefs Biological Soil Crusts.
It fixes the sand compared to physics, the cultivation of chemical sand-fixing and vegetation, in such a way that microorganism skinning fixes the sand as novel fix the sand, has suitable
The advantages such as Ying Xingqiang, at low cost, quick.Present invention firstly discloses to molten bacillus fix the sand ability qualitative, quantitative research.
It is micro- that molten bacillus (Lysobacter sp.) SCSIO 17111 of the invention was preserved in China on August 18th, 2017
Biological inoculum preservation administration committee common micro-organisms center (CGMCC), address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Number, Institute of Microorganism, Academia Sinica, deposit number: CGMCC No.14537.
Detailed description of the invention:
Fig. 1 is the effect of fixing the sand that bacterial strain SCSIO 17111 is less than 1.25mm and the sterile coral sand greater than 0.2mm to partial size
Fruit.
Fig. 2 is the sand-fixing effect of sterile coral sand of the bacterial strain SCSIO 17111 to partial size less than 0.2mm.
Specific embodiment:
The following examples are further illustrations of the invention, rather than limiting the invention.
Embodiment 1: the separation of molten bacillus (Lysobacter sp.) SCSIO 17111
1. sample acquires
Biological Soil Crusts sample picks up from South China Sea islands and reefs in November, 2016.After sample acquisition, it is quickly charged with sterile
Polyethylene sampler bag in, dry in the shade, be stored in room temperature.
2. isolation medium
Isolation medium be TSA (Tryptose soy agar) culture medium: commercial product (BD), tryptone 15.0g/L, greatly
Legumin peptone 5.0g/L, sodium chloride 5.0g/L, agar 15.0g/L, solvent is water, pH7.3 ± 0.2.Its preparation method is will be upper
It states component to be uniformly mixed by its content, adjusts pH value to pH7.3 ± 0.2, sterilization to obtain the final product.
3. the separation screening of bacterial strain
Skinning sample is fully ground and is mixed, 2g is then taken to be placed in the conical flask equipped with 18ml aseptic deionized water
In, be placed with some sterilized small beades in conical flask, 30 DEG C, 200r/min oscillation shake up 30min, taken after standing 1min
The sample suspension of 1ml carries out gradient dilution, take respectively 10 times of 200 μ l stostes and dilution, 100 times, 1000 times, 10000 times,
100000 times, 1000000 times of sample be coated on isolation medium, each processing sets 3 repetitions.30 DEG C are cultivated 2-4 days, root
According to the form of bacterium colony, single colonie is chosen from plate, and carries out four ride purifying on fresh culture medium.Thus it separates pure
Change and obtains bacterial strain SCSIO 17111.
Gene is carried out using EasyPure Bacteria Genomic DNAKit (Quan Shijin) to bacterial strain SCSIO 17111
Group DNA extracting, then expands 16S rRNA genetic fragment with universal primer 27F/1492R and is sequenced, sequence such as SEQ ID
Shown in No.1.Pass through the online comparison of the website EZBioCloud to 16S rRNA gene order, finds nearest with its affiliation
Be bacterial strain PB-6250T, the entitled Lysobacterfirmicutimachus of strain, similarity 99.78%.Therefore, this hair
Bright bacterial strain SCSIO 17111 belongs to molten bacillus Lysobacter and belongs to, and is named as molten bacillus (Lysobacter sp.)
SCSIO 17111, the bacterium were preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms on August 18th, 2017
The heart (CGMCC), address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica, deposit number:
CGMCC No.14537。
Molten bacillus (Lysobacter sp.) SCSIO 17111 of the invention with it is in the prior art
Lysobacterfirmicutimachus PB-6250TWith many differences, such as Lysobacterfirmicutimachus
PB-6250TIt does not grow at 42 DEG C, and can be grown at 10 DEG C without record, but molten bacillus (Lysobacter of the invention
Sp.) SCSIO 17111 can be grown at 42 DEG C and 10 DEG C, while molten bacillus (Lysobacter sp.) SCSIO of the invention
17111 optimum growth temperature is 28-32 DEG C, and Lysobacterfirmicutimachus PB-6250TThe most suitable growth temperature
Degree is 25-28 DEG C.L.firmicutimachus PB-6250TIt is only capable of growing when NaCl concentration (w/v) is less than or equal to 0.5%,
And molten bacillus (Lysobacter sp.) SCSIO 17111 of the invention can be grown when NaCl concentration (w/v) is 1%；In addition,
Have no that Lysobacter belongs to known kind of sand-fixing effect related introduction at present, and this new strains of the invention have sand-fixing effect,
Therefore molten bacillus (Lysobacter sp.) SCSIO 17111 of the invention is the new strain that Lysobacter belongs to.
Embodiment 2: the measurement of molten 17111 yield of extracellular polysaccharide of bacillus SCSIO
Molten bacillus SCSIO 17111 is inoculated in TSB culture medium (BD), medium component is tryptone 17.0g/L,
Soy peptone 3.0g/L, glucose 2.5g/L, sodium chloride 5.0g/L, K2HPO42.5g/L, solvent are water, pH7.3 ± 0.2,
Preparation method is to be uniformly mixed mentioned component by its content, adjusts pH value, then sterilizes again to obtain the final product.30 DEG C, 180r/min condition
Under, cultivate 45h.Culture solution is removed into thallus through 11603g centrifugation 20min, then supernatant is passed through to 0.22 μm of filter membrane mistake again
Filter, the sufficiently remaining microbial cell of removal.Then filtrate is poured into test tube, 95% second of volume fraction of 4 times of volumes is added
Alcohol solution, 4 DEG C of precipitates overnights.Then mixed liquor is centrifuged 20min through 2057g, removes supernatant, collect sediment.It is obtained
Sediment successively washed through acetone, dehydrated alcohol, precipitating after washing and then add the trichlorine that volume fraction is 80%
Acetic acid 4ml, for removing isolating protein.Finally liquid is removed, puts the precipitate in 60 DEG C of dry 30h in baking oven, is obtained extracellular
Polysaccharide sample.
Measurement of the polysaccharide content is carried out to exocellular polysaccharide sample using phend-sulphuric acid.Monosaccharide standard curve is made first:
Accurately weighing glucose (analysis is pure), 10mg in 50ml beaker, adds deionized water sufficiently to dissolve, and aqueous solution is poured into 250ml
In volumetric flask, add deionized water constant volume, draws 0.2ml, 0.3ml, 0.4ml, 0.5ml, 0.6ml, 0.7ml, 0.8ml respectively in examination
Guan Zhong, each moisturizing to 1.0ml.Then the 6% phenol 0.5ml and concentrated sulfuric acid (analysis is pure, 95.5%) 2.5ml is added, stands
10min shakes up, and is placed at room temperature for after 20min and surveys absorbance in 490nm with spectrophotometer, with 1.0ml deionized water by same aobvious
Color operation is blank control.Abscissa is glucose quality, and ordinate is absorbance value, draws standard curve.It then will preparation
The exocellular polysaccharide sample of molten bacillus SCSIO 17111 be redissolved in the deionized water isometric with former strain cultured solution, system
At solution.It is put into test tube from 0.8ml is pipetted in sample solution, then plus water is mended to 1.0ml.Then 6% phenol 0.5ml is added
And the concentrated sulfuric acid (analysis is pure, 95.5%) 2.5ml, 10min is stood, is shaken up, is existed after being placed at room temperature for 20min using spectrophotometer
Absorbance is surveyed under 490nm, same color operation is pressed as blank control using 1.0ml deionized water, measures absorbance, according to standard song
Line computation goes out polyoses content.Testing result shows that the molten TSB culture medium culture 45h of bacillus SCSIO 17111, exocellular polysaccharide are produced
Amount is 0.08 ± 0.01mg/ml.
Embodiment 3: molten bacillus SCSIO 17111 fixes the sand the measurement of ability
Coral sand is successively crossed into 16 mesh (sieve pore 1.25mm) and 80 mesh (sieve pore 0.20mm) sifter device, partial size is obtained respectively and is less than
It, is loaded on different culture dishes by 1.25mm and the coral sand of coral sand and partial size less than 0.20mm for being greater than 0.20mm respectively
In, every kind of specification is sandy to do 3 repetitions, then by its 121 DEG C sterilizing 25min, is placed in baking oven and dries and mix to pine for 60 DEG C
It dissipates uniform.Molten bacillus SCSIO 17111 is inoculated in TSB culture medium (with embodiment 2), 30 DEG C, under the conditions of 180r/min, vibration
After swinging culture 48h, by bacterium solution loaded in sterile small watering can, uniformly it is sprayed on coral sand, is with the TSB culture medium of sterilizing
Blank control.In 30 DEG C, after culture 55 days, the thickness of skinning is measured.Testing result shows molten bacillus SCSIO 17111
The formation that can effectively promote skinning, when culture solution is uniformly sprayed at partial size less than 1.25mm and is greater than the sterile of 0.20mm
When on coral sand, the skinning of formation is with a thickness of 6.00 ± 1.32mm (Fig. 1)；It is less than when culture solution is uniformly sprayed at partial size
When on the sterile coral sand of 0.20mm, the skinning of formation is with a thickness of 2.33 ± 0.58mm (Fig. 2).Although the skinning thickness of the latter is not
Such as the former, but the soil aggregation object stability of the latter is stronger, i.e., when carrying out dry screen to soil aggregation object, the aggregate energy of the latter
Preferable stability is kept, and the former is more easily broken.And after the TSB culture medium as control is sprayed onto coral sand, two kinds of partial sizes
The coral sand of size is all without skinning.It can be seen that molten bacillus SCSIO 17111 provided by the present invention can make the grains of sand reunite
And maintain a relatively stable state, to play the effect fixed the sand, can be applied to prevention and treatment arid, semiarid zone desertification and
The building of islands and reefs Biological Soil Crusts.It fixes the sand compared to physics, the cultivation of chemical sand-fixing and vegetation, is fixed the sand using microorganism skinning
As novel mode of fixing the sand, the adaptable advantages such as strong, at low cost, quick.
Sequence table
<110>Chinese Academy of Science Nanhai Ocean Research Institute
The molten bacillus SCSIO 17111 and its application that<120>one plants of extracellular polysaccharide can fix the sand
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1390
<212> DNA
<213>molten bacillus SCSIO 17111 (Lysobacter sp.)
<400> 1
atgcagtcga acggcagcac agaggagctt gctccttggg tggcgagtgg cggacgggtg 60
aggaatacgt cggaatctgc ctatttgtgg gggataacgt agggaaactt acgctaatac 120
cgcatacgac ctacgggtga aagtggggga ccgcaaggcc tcacgcagat agatgagccg 180
acgtcggatt agctagttgg cggggtaaag gcccaccaag gcgacgatcc gtagctggtc 240
tgagaggatg atcagccaca ctggaactga gacacggtcc agactcctac gggaggcagc 300
agtggggaat attggacaat gggcgcaagc ctgatccagc catgccgcgt gtgtgaagaa 360
ggccttcggg ttgtaaagca cttttgtccg gaaagaaaag cttagggtta ataaccttga 420
gtcatgacgg taccggaaga ataagcaccg gctaacttcg tgccagcagc cgcggtaata 480
cgaagggtgc aagcgttact cggaattact gggcgtaaag cgtgcgtagg tggtttgtta 540
agtctgatgt gaaagccctg ggctcaacct gggaatggca ttggaaactg gcttactaga 600
gtgcggtaga gggtagcgga attcccggtg tagcagtgaa atgcgtagat atcgggagga 660
acatccgtgg cgaaggcggc tacctggacc agcactgaca ctgaggcacg aaagcgtggg 720
gagcaaacag gattagatac cctggtagtc cacgccctaa acgatgcgaa ctggatgttg 780
ggggcaactt ggccctcagt atcgaagcta acgcgttaag ttcgccgcct gggaagtacg 840
gtcgcaagac tgaaactcaa aggaattgac gggggcccgc acaagcggtg gagtatgtgg 900
tttaattcga tgcaacgcgc agaaccttac ctggccttga catccacgga actttccaga 960
gatggattgg tgccttcggg aaccgtgaga caggtgctgc atggctgtcg tcagctcgtg 1020
tcgtgagatg ttgggttaag tcccgcaacg agcgcaaccc ttgtccttag ttgccagcac 1080
gtaatggtgg gaactctaag gagaccgccg gtgacaaacc ggaggaaggt ggggatgacg 1140
tcaagtcatc atggccctta cggccagggc tacacacgta ctacaatggt agggacagag 1200
ggctgcaaac ccgcgagggc aagccaatcc cagaaaccct atctcagtcc ggattggagt 1260
ctgcaactcg actccatgaa gtcggaatcg ctagtaatcg cagatcagca ttgctgcggt 1320
gaatacgttc ccgggccttg tacacaccgc ccgtcacacc atgggagttt gttgcaccag 1380
aagcaggtag 1390
Claims (4)
1. molten bacillus (Lysobacter sp.) SCSIO 17111, deposit number: CGMCC No.14537.
2. molten bacillus SCSIO 17111 described in claim 1 is fixing the sand and/or is establishing the application in Biological Soil Crusts.
3. application according to claim 2, which is characterized in that be the bacterium solution for first cultivating molten bacillus SCSIO 17111, then
Bacterium solution is inoculated into the region for needing to fix the sand.
4. application according to claim 3, which is characterized in that the bacterium solution of the described molten bacillus SCSIO 17111 of culture is
With the molten bacillus SCSIO 17111 of TSB culture medium culture, bacterium solution, the TSB culture medium are obtained are as follows: tryptone 17.0g/L,
Soy peptone 3.0g/L, glucose 2.5g/L, sodium chloride 5.0g/L, K2HPO42.5g/L, solvent are water, pH7.3 ± 0.2.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810609500.3A CN108795807B (en) | 2018-06-13 | 2018-06-13 | A strain of Lysobacter SCSIO17111 that produces exopolysaccharide and can fix sand and its application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810609500.3A CN108795807B (en) | 2018-06-13 | 2018-06-13 | A strain of Lysobacter SCSIO17111 that produces exopolysaccharide and can fix sand and its application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108795807A CN108795807A (en) | 2018-11-13 |
| CN108795807B true CN108795807B (en) | 2019-06-21 |
Family
ID=64085768
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810609500.3A Active CN108795807B (en) | 2018-06-13 | 2018-06-13 | A strain of Lysobacter SCSIO17111 that produces exopolysaccharide and can fix sand and its application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108795807B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111117892A (en) * | 2020-01-17 | 2020-05-08 | 北京大学 | Auxenochlorella sp. BSC-01 and its application |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113287382B (en) * | 2021-05-20 | 2022-08-05 | 中国科学院南海海洋研究所 | Algal fungus skinning cultivation method suitable for calcareous sandy soil of island reef of south China sea coral |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103667136A (en) * | 2013-12-10 | 2014-03-26 | 南京农业大学 | Separation and identification of lysobacter antibioticus OH13 capable of antagonizing plant pathogenic bacteria |
| CN105886424A (en) * | 2016-03-02 | 2016-08-24 | 云南瑞升烟草技术（集团）有限公司 | Lysobacter capsici having erysiphe cichoracearum resistance and application of lysobacter capsici |
| CN106434440A (en) * | 2016-09-21 | 2017-02-22 | 云南农业大学 | Lysobacter capsici ZST1-2 and application thereof |
| CN106434439A (en) * | 2016-09-21 | 2017-02-22 | 云南农业大学 | Lysobacter enzymogenes 1-T-1-4 and application thereof |
| CN107099484A (en) * | 2017-06-26 | 2017-08-29 | 西南大学 | One plant of molten bacillus of producing enzyme and its application |
-
2018
- 2018-06-13 CN CN201810609500.3A patent/CN108795807B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103667136A (en) * | 2013-12-10 | 2014-03-26 | 南京农业大学 | Separation and identification of lysobacter antibioticus OH13 capable of antagonizing plant pathogenic bacteria |
| CN105886424A (en) * | 2016-03-02 | 2016-08-24 | 云南瑞升烟草技术（集团）有限公司 | Lysobacter capsici having erysiphe cichoracearum resistance and application of lysobacter capsici |
| CN106434440A (en) * | 2016-09-21 | 2017-02-22 | 云南农业大学 | Lysobacter capsici ZST1-2 and application thereof |
| CN106434439A (en) * | 2016-09-21 | 2017-02-22 | 云南农业大学 | Lysobacter enzymogenes 1-T-1-4 and application thereof |
| CN107099484A (en) * | 2017-06-26 | 2017-08-29 | 西南大学 | One plant of molten bacillus of producing enzyme and its application |
Non-Patent Citations (1)
| Title |
|---|
| 溶杆菌属及其在植物病害防治中的研究进展;姬广海;《云南农业大学学报》;20111231;第26卷(第1期);124-130 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111117892A (en) * | 2020-01-17 | 2020-05-08 | 北京大学 | Auxenochlorella sp. BSC-01 and its application |
| CN111117892B (en) * | 2020-01-17 | 2020-09-08 | 北京大学 | Green alga Auxenochlorella sp.BSC-01 and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108795807A (en) | 2018-11-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US20200063109A1 (en) | Phage composition and use thereof in inactiving antibiotic resistance pathogenic bacteria | |
| CN106754515B (en) | Screening method of growth promoting strains for improving salt tolerance of crops | |
| CN113373094B (en) | Brevibacterium fritolerans SDB5 and application thereof in promoting plant growth | |
| CN107090419B (en) | Bacillus amyloliquefaciens and its application | |
| CN104560827B (en) | A kind of biocontrol actinomycetes bacterial strain for preventing and treating tobacco bacterial wilt and its application | |
| CN103667132A (en) | Bacillus cereus and application thereof to improve plant drought-resistant capability | |
| CN111187743B (en) | Bacillus thuringiensis strain YN-2-2 and application thereof | |
| CN108795807B (en) | A strain of Lysobacter SCSIO17111 that produces exopolysaccharide and can fix sand and its application | |
| CN109706125B (en) | Bacillus cereus phage composition and application thereof | |
| CN112438277B (en) | Xylariaceae fungus sj18 and application thereof in prevention and treatment of pine wood nematodes | |
| CN110846262B (en) | A Serratia marcescens strain SZ201 and its application | |
| CN110358695A (en) | A kind of drought resisting growth promoting bacteria agent and its screening technique, liquid bacterial agent | |
| CN111004736B (en) | A kind of Bacillus megaterium and its application of degrading pyrethroid insecticides | |
| CN110408567B (en) | A Salt-Alkali-tolerant Methylotrophic Bacillus and its Viable Preparation and Application | |
| CN116333928A (en) | Separation and application of suaeda salsa rhizosphere growth-promoting bacteria | |
| CN114480179B (en) | Bacteria for separating saline-alkali soil and application thereof | |
| CN116083241A (en) | Separation and purification method and application of plant rhizosphere growth promoting bacteria | |
| CN116179398A (en) | Bacillus belicus JSBV55, biocontrol microbial inoculum, preparation method and application thereof, and control method of garlic purple spot | |
| CN109182194A (en) | One plant of Yang Ling rhizobium for promoting coronule flower growth and its cultural method and application | |
| CN105087450B (en) | The exocellular polysaccharide of one plant of marine bacteria and its generation | |
| CN115305222B (en) | A kind of Bacillus strain and its application | |
| CN117887638A (en) | Tricholoma giganteum growth-promoting microorganism SLTmB04 strain and application thereof | |
| CN105695355A (en) | Method for preparing bacterium-algae cells through two-wheel coculturing and flocculation method | |
| CN114525223B (en) | A Pseudomonas putida and its application in degrading potato starch wastewater | |
| CN114058550A (en) | Growth-promoting synthetic flora for efficient colonization of tobacco root surface and screening method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |