CN106119168A - One strain has rich phosphorus, degrading organic phosphor and the Serratieae of suppression plant pathogenic fungi - Google Patents
One strain has rich phosphorus, degrading organic phosphor and the Serratieae of suppression plant pathogenic fungi Download PDFInfo
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- CN106119168A CN106119168A CN201610580077.XA CN201610580077A CN106119168A CN 106119168 A CN106119168 A CN 106119168A CN 201610580077 A CN201610580077 A CN 201610580077A CN 106119168 A CN106119168 A CN 106119168A
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- serratieae
- serratia
- mew06
- pathogenic fungi
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- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 title claims abstract description 48
- 229910052698 phosphorus Inorganic materials 0.000 title claims abstract description 37
- 239000011574 phosphorus Substances 0.000 title claims abstract description 37
- 244000000004 fungal plant pathogen Species 0.000 title claims abstract description 14
- 230000001629 suppression Effects 0.000 title claims abstract description 14
- 230000000593 degrading effect Effects 0.000 title claims abstract description 13
- 230000001580 bacterial effect Effects 0.000 claims abstract description 26
- 239000005562 Glyphosate Substances 0.000 claims abstract description 19
- MCWXGJITAZMZEV-UHFFFAOYSA-N dimethoate Chemical compound CNC(=O)CSP(=S)(OC)OC MCWXGJITAZMZEV-UHFFFAOYSA-N 0.000 claims abstract description 19
- XDDAORKBJWWYJS-UHFFFAOYSA-N glyphosate Chemical compound OC(=O)CNCP(O)(O)=O XDDAORKBJWWYJS-UHFFFAOYSA-N 0.000 claims abstract description 19
- 229940097068 glyphosate Drugs 0.000 claims abstract description 19
- 241000894006 Bacteria Species 0.000 claims description 27
- 239000004576 sand Substances 0.000 claims description 3
- 230000015556 catabolic process Effects 0.000 abstract description 21
- 238000006731 degradation reaction Methods 0.000 abstract description 21
- 239000003987 organophosphate pesticide Substances 0.000 abstract description 11
- 230000000813 microbial effect Effects 0.000 abstract description 5
- 238000005516 engineering process Methods 0.000 abstract description 3
- 244000053095 fungal pathogen Species 0.000 abstract description 2
- 230000002401 inhibitory effect Effects 0.000 abstract description 2
- 241000607714 Serratia sp. Species 0.000 description 20
- 239000000575 pesticide Substances 0.000 description 16
- 238000000034 method Methods 0.000 description 13
- 241000123650 Botrytis cinerea Species 0.000 description 8
- 238000000855 fermentation Methods 0.000 description 8
- 230000004151 fermentation Effects 0.000 description 8
- 239000001963 growth medium Substances 0.000 description 8
- 239000002609 medium Substances 0.000 description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 8
- 241000607715 Serratia marcescens Species 0.000 description 7
- 108020004465 16S ribosomal RNA Proteins 0.000 description 6
- 208000031481 Pathologic Constriction Diseases 0.000 description 6
- 210000001215 vagina Anatomy 0.000 description 6
- 239000002351 wastewater Substances 0.000 description 6
- 230000000694 effects Effects 0.000 description 5
- 239000002054 inoculum Substances 0.000 description 5
- 244000052769 pathogen Species 0.000 description 5
- 230000001717 pathogenic effect Effects 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 241000832823 Serratia nematodiphila Species 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 238000005070 sampling Methods 0.000 description 4
- 210000000582 semen Anatomy 0.000 description 4
- 238000010186 staining Methods 0.000 description 4
- 238000013456 study Methods 0.000 description 4
- 241000813090 Rhizoctonia solani Species 0.000 description 3
- 230000003321 amplification Effects 0.000 description 3
- 238000004737 colorimetric analysis Methods 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 229910052750 molybdenum Inorganic materials 0.000 description 3
- 239000011733 molybdenum Substances 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- 239000002689 soil Substances 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- ZOKXTWBITQBERF-UHFFFAOYSA-N Molybdenum Chemical compound [Mo] ZOKXTWBITQBERF-UHFFFAOYSA-N 0.000 description 2
- 240000007594 Oryza sativa Species 0.000 description 2
- 235000007164 Oryza sativa Nutrition 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- XYFCBTPGUUZFHI-UHFFFAOYSA-N Phosphine Chemical compound P XYFCBTPGUUZFHI-UHFFFAOYSA-N 0.000 description 2
- 241000607720 Serratia Species 0.000 description 2
- 241000609666 Tuber aestivum Species 0.000 description 2
- 241000607479 Yersinia pestis Species 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 229960004373 acetylcholine Drugs 0.000 description 2
- 239000003905 agrochemical Substances 0.000 description 2
- 230000000843 anti-fungal effect Effects 0.000 description 2
- 229940121375 antifungal agent Drugs 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 239000012531 culture fluid Substances 0.000 description 2
- 239000003344 environmental pollutant Substances 0.000 description 2
- 229940088598 enzyme Drugs 0.000 description 2
- 210000003495 flagella Anatomy 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 230000002538 fungal effect Effects 0.000 description 2
- 230000028644 hyphal growth Effects 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 230000001788 irregular Effects 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 239000005645 nematicide Substances 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 231100000719 pollutant Toxicity 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 1
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 1
- 241000588624 Acinetobacter calcoaceticus Species 0.000 description 1
- 241001530056 Athelia rolfsii Species 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000186146 Brevibacterium Species 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 244000025254 Cannabis sativa Species 0.000 description 1
- 102000012286 Chitinases Human genes 0.000 description 1
- 108010022172 Chitinases Proteins 0.000 description 1
- 102000003914 Cholinesterases Human genes 0.000 description 1
- 108090000322 Cholinesterases Proteins 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 108090000371 Esterases Proteins 0.000 description 1
- 239000007836 KH2PO4 Substances 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000588772 Morganella morganii Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 102100021079 Ornithine decarboxylase Human genes 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 230000000895 acaricidal effect Effects 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- OIPILFWXSMYKGL-UHFFFAOYSA-N acetylcholine Chemical compound CC(=O)OCC[N+](C)(C)C OIPILFWXSMYKGL-UHFFFAOYSA-N 0.000 description 1
- -1 acetylcholine ester Chemical class 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 229910052787 antimony Inorganic materials 0.000 description 1
- WATWJIUSRGPENY-UHFFFAOYSA-N antimony atom Chemical compound [Sb] WATWJIUSRGPENY-UHFFFAOYSA-N 0.000 description 1
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 description 1
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 1
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 1
- 238000010170 biological method Methods 0.000 description 1
- 230000033558 biomineral tissue development Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 229940048961 cholinesterase Drugs 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000004064 dysfunction Effects 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 235000014103 egg white Nutrition 0.000 description 1
- 210000000969 egg white Anatomy 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 229910052564 epsomite Inorganic materials 0.000 description 1
- 238000012851 eutrophication Methods 0.000 description 1
- 238000009313 farming Methods 0.000 description 1
- 239000003337 fertilizer Substances 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 230000002363 herbicidal effect Effects 0.000 description 1
- 239000004009 herbicide Substances 0.000 description 1
- BHEPBYXIRTUNPN-UHFFFAOYSA-N hydridophosphorus(.) (triplet) Chemical compound [PH] BHEPBYXIRTUNPN-UHFFFAOYSA-N 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000000749 insecticidal effect Effects 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 229940076266 morganella morganii Drugs 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000003986 organophosphate insecticide Substances 0.000 description 1
- 238000011056 performance test Methods 0.000 description 1
- 230000000361 pesticidal effect Effects 0.000 description 1
- 150000003014 phosphoric acid esters Chemical group 0.000 description 1
- 229910000073 phosphorus hydride Inorganic materials 0.000 description 1
- 238000013081 phylogenetic analysis Methods 0.000 description 1
- 238000000053 physical method Methods 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000009418 renovation Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000004062 sedimentation Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- RYYWUUFWQRZTIU-UHFFFAOYSA-K thiophosphate Chemical compound [O-]P([O-])([O-])=S RYYWUUFWQRZTIU-UHFFFAOYSA-K 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000033 toxigenic Toxicity 0.000 description 1
- 230000001551 toxigenic effect Effects 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
Classifications
-
- 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
- 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
- C12R2001/425—Serratia
Abstract
The invention discloses a strain and there is rich phosphorus, degrading organic phosphor and the Serratieae of suppression plant pathogenic fungi, belong to microbial technology field.This bacterial strainSerratiaSp. MEW06 is preserved in China typical culture collection center, deposit number is CCTCC NO. M 2015779, the Serratieae of the present invention has the rich phosphorus ability of transitory efficient, can be used for preparing bio-feritlizer, also the pathogenic fungi of crop is had inhibitory effect, can be applicable to microbial control, simultaneously, this bacterial strain has good Degradation to organophosphorus pesticide glyphosate and Rogor, can effectively remove organophosphorus pesticide pollution, and the Serratieae of the present invention is with a wide range of applications.
Description
Technical field
The invention belongs to microbial technology field, be specifically related to Serratieae MEW06 and this bacterial strain thereof in rich phosphorus, degraded
Application in terms of organophosphor and suppression plant pathogenic fungi.
Background technology
Organophosphorus pesticide is the big classification of in current pesticide, accounts for about the 80% of Pesticidal products, because it is to preventing and treating agricultural
Disease pest and weed have economy, efficiently, the feature such as quickly, produce the most in a large number and be widely used.But, due to organic
The expanding production of phosphorus pesticide chemical factory, the wastewater flow rate discharged in its production process is big, and COD content is high, and pollutant component is complicated,
Toxicity is big, difficult for biological degradation, and environment causes harm greatly.And can spread from applying area to surrounding after the applications of pesticide,
In soil, accumulation migrates, and enters water body or plant etc., and by the enrichment of food chain, human security is produced hidden danger.Organophosphor
Pesticide mostly is phosphoric acid ester or group thiophosphate, can suppress cholinesterase activity so that it is cannot decompose phatidylcholine, cause god
Through dysfunction, severe patient may cause death；Secondly, part organophosphorus pesticide is become CO by microbial mineralization in environment2And nothing
After machine phosphorus, cause content of inorganic phosphorus in water body to raise, cause the eutrophication of water body.The uncontrolled production of pesticide chemical factory and
Pesticide is irrational to be widely used, and causes increasingly severe consequence to the mankind and ecological environment, and the pollution problem of pesticide is
Become the focus of global concern.Scientist has carried out the work of many correlational studyes both at home and abroad, has had the degradation technique of pesticide relatively
Great development, including various physics, chemistry, biological method etc..High with traditional physical method and chemical method processing cost, may
Produce new pollutant to compare, the biological renovation method utilizing microorganism and biological product thereof to carry out degradation of contaminant have nontoxic,
The advantage such as noresidue, non-secondary pollution, be eliminate and removing toxic substances agricultural chemicals waste water a kind of safely, effectively, cheap method.
Rogor is interior absorption organophosphor Insecticidal and acaricidal agent, it is adaptable to preventing and treating sucking pest, and purposes is wide, yield is big,
But Rogor is volatile to be gathered in soil to air, water body and sedimentation, pollute farming animals fishing side-product, and pass through food chain transport
To human body, people is produced harm.Glyphosate is the steriland herbicide of a kind of high-efficiency low-toxicity, inner sucting conduction type, owing to not having
Selectivity, its range of application the most progressively expands.But in glyphosate production wastewater, chemical residual is big, and environmental pollution is the tightest
Weight.Therefore, the bioremediation of research Rogor and glyphosate seems extremely necessary.
The Rogor degradation bacteria being separated at present mainly has acinetobacter calcoaceticus, Bacillus cercus, Pseudomonas aeruginosa, spore bar
Bacterium etc.；Glyphosate degradation bacterium mainly has pseudomonas, morganella morganii etc.；Existing pesticide degradation bacteria is happy to organophosphorus insecticide
The degradation efficiency of fruit and glyphosate is low, bacterial strain function singleness, and actual application is limited.The Serratieae of the present invention can be with efficient degradation
Glyphosate and Rogor, it is possible to absorption inorganic phosphorus in wastewater, the improvement being applied to pollute environment has important reality meaning
Justice and application prospect.Currently, with respect to serratia marcescens high yield chitinase and the red non-dispersive pigment-spirit bar of secretion thereof
Rhzomorph has antibacterial, regulates immunologic function, presses down the functions such as cancer and report more both at home and abroad, but have Rogor and glyphosate degradation effect
Serratieae have no report.
Summary of the invention
It is an object of the invention to the practical problem according to current organophosphorus pesticide pollution and needs, it is provided that a strain has richness
Phosphorus, degrading organic phosphor pesticides and the Serratieae of suppression plant pathogenic fungi.
The present invention has rich phosphorus, degrading organic phosphor pesticides and the Serratieae Serratia of suppression plant pathogenic fungi
Sp.MEW06 is in China typical culture collection center preservation (Wuhan, China Wuhan University), and deposit number is CCTCC NO.M
2015779, preservation date is December in 2015 25.
Bacterium colony and the thalline of bacterial strain of the present invention are characterized as: cultivating at LB solid medium, bacterium colony is cerise, surface light
Sliding, moistening, opaque, edge is irregular, and size is 0.9-1.2 μ m 1.5-2.0 μm, and during 28 DEG C of cultivations, bacterium solution presents scarlet
Color, 37 DEG C are not developed the color when cultivating.Observe and spore staining, flagella staining, capsule stain and Gram’s staining through SEM, send out
Existing MEW06 bacterial strain is subsphaeroidal brevibacterium, do not produce spore, peritrichous, without pod membrane, Gram-negative.
The molecular biological variety identification method of bacterial strain of the present invention is: with Serratia sp.MEW06 genome as template, uses
Universal primer (27F and 1492R), amplification 16S rDNA sequence (see sequence table), MEW06 16S rDNA PCR extension increasing sequence warp
NCBI Blast carries out homology comparison analysis, find bacterial strain MEW06 and Serratia marcescens (GenBank:
KT894728) sequence similarity is 99%, with the sequence of Serratia nematodiphila (GenBank:KC172028)
Similarity is also 99%.Serratia sp.MEW06 16S rDNA sequence evolve on and serratia marcescens (Serratia
Marcescens) close with addicted to nematicide Serratieae (Serratia nematodiphila).
What the present invention provided has rich phosphorus, degrading organic phosphor pesticides and suppression plant pathogenic fungi Serratieae by domestication
Method obtains, and comprises the following steps: gathers Sand lake and the water sample of periphery thereof, takes 5-10mL and be inoculated in LB culture fluid, enrichment
Cultivate；Then bacteria suspension is inoculated in the fermentation medium containing pesticide, passage and attenuation 5 times by the inoculum concentration (V/V) of 5%-10%
After, line LB solid medium, according to the generation situation of bacterium colony on flat board, the well-grown bacterium colony of picking, be inoculated in containing agriculture
The fermentation medium of medicine, uses acetylcholine ester enzyme inhibition rate method (GB/T5009.199-2003) to measure organophosphorus pesticide and contains
Amount, filters out the bacterial strain Serratia sp.MEW06 higher to degradation of pesticide efficiency, and by molybdenum blue colorimetric method, flat board stands facing each other
Method measures rich phosphorus ability and suppression plant pathogenic fungi ability respectively.
The bacterial strain Serratia sp.MEW06 of separation screening is carried out performance test:
A. the mensuration of strains for degrading organophosphorus pesticide ability
For inquiring into the aimed strain Serratia sp.MEW06 degradation capability to organophosphorus pesticide, choose agricultural at present raw
Organophosphorus pesticide Rogor conventional on product and glyphosate are as target pesticide.By screening Serratia sp.MEW06 after purification
Bacterial strain, activated overnight, regulate bacterium solution OD600=1.0, by the inoculum concentration of 4% be forwarded to containing 50mg/L, 100mg/L, 150mg/L,
200mg/L glyphosate or the fermentation culture of Rogor, 150rpm, 28 DEG C of cultivations, in 1d, 3d, 5d, 7d sampling and measuring degradation rate.
B. the mensuration of bacterial strain richness phosphorus ability
By screening Serratia sp.MEW06 bacterial strain after purification, the inoculum concentration by 2% is inoculated in phosphorus content respectively and is
In the PAM culture medium of 10mg/L, 20mg/L, in 0h, 6h, 12h, 24h sampling and measuring poly-phosphorus rate.It is centrifuged 10min through 6000r/min
After, take supernatant and measure content of inorganic phosphorus (GB 11893-89-molybdenum antimony resistance colorimetric method), with the enrichment culture liquid before access bacterial strain
Inorganic phosphorus concentration compare, calculate poly-phosphorus rate.
The mensuration of c.MEW06 bacterial strain antifungal ability
Take the eugonic sharp knife Fusariumsp of a diameter of 5mm, Fructus Mali pumilae wheel stricture of vagina bacterium, Semen arachidis hypogaeae leaf mold, Rhizoctonia solani, Fructus Fragariae Ananssae
Grey mold and the pathogen of Botrytis cinerea truffle are inoculated in the central authorities of PDA plate, cultivate 2d for 28 DEG C.Put in the position of distance mycelia edge 2cm
Put sterilized filter paper (D=6mm), bacterium solution and the PBS (blank) of 60 μ L are dropped on filter paper respectively.28 DEG C of continuation
After cultivating 2 days, observe the test sample suppression situation to Fungal hyphal growth.
Compared with prior art, present invention have the advantage that
1, the bacterial strain Serratia sp.MEW06 of the present invention is strong to the degradation capability of Rogor, glyphosate, and does not cause solution
Middle phosphorus content raises, and can grow under extremely low nutrient environment, and resistance ability is high, therefore can be applicable to the biology of agricultural chemicals waste water
Processing, this bacterial strain has and presses down pathogenic fungi ability simultaneously, to sharp knife Fusariumsp, Fructus Mali pumilae wheel stricture of vagina bacterium, Semen arachidis hypogaeae leaf mold, rice banded sclerotial blight
Bacterium, Botrytis cinerea and the pathogen of Botrytis cinerea have preferably inhibition, and Serratia sp.MEW06 can be at Biological control, micro-
The field such as bio-feritlizer, water harnessing is applied, and has wide market prospect.
2, this bacterial strain Serratia sp.MEW06 amplification culture method is simple, and toxigenic capacity is low, can with large-scale culture,
It is prepared as in bio-feritlizer or microbial bacterial agent degraded soil residual pesticide or removes phosphorous chemical industry enterprise organophosphor and Phos
Pollute.
Accompanying drawing explanation
Fig. 1 is Serratia sp.MEW06 colonial morphology (A), spore staining (B) and scanning electron microscope (C) figure.
Fig. 2 is organic phosphorus degrading rate ((A): Rogor, (B): grass is sweet over time of Serratia sp.MEW06
Phosphine).
Fig. 3 is Serratia sp.MEW06 antifungal lab diagram, (A) sharp knife Fusariumsp；(B) Fructus Mali pumilae wheel stricture of vagina bacterium；(C) flower
Leave is mould；(D) Rhizoctonia solani；(E) Botrytis cinerea；(F) the pathogen of Botrytis cinerea；A (), (c), (e), (g), (i) and (k) are blank right
According to PBS；(b), (d), (f), (h), (j) and (l) MEW06 fermentation liquid.
It is embodied as case
The following is the specific embodiment of the present invention, technical scheme is described further, but the present invention's is interior
Hold the scope being not limited solely to described in embodiment, every change without departing substantially from present inventive concept or equivalent replacement and be included in this
Within the protection domain of invention.
Embodiment 1
The water sample gathering Sand lake and periphery thereof is added to LB culture medium, 28 DEG C of 150r/min enrichment culture 24h.By upper
State bacteria suspension and be inoculated in the fermentation medium containing glyphosate 200mg/L, 28 DEG C of 150r/min enrichments by the inoculum concentration of 5%-10%
After cultivating 3d, being forwarded to same medium, continue under the same conditions to cultivate, during passage and attenuation, glyphosate content progressively carries
Height arrives as 1000mg/L；The bacterium solution passing on 5 times is lined LB solid medium, 28 DEG C, cultivates 24-48h, according in culture medium
The generation situation of bacterium colony, the well-grown colony inoculation of picking in the LB culture fluid (diluting 5 times) containing glyphosate 50mg/L, 28 DEG C
150r/min cultivates 72h, uses acetylcholine enzyme inhibition rate method (GB/T5009.199-2003) to measure organophosphorus pesticide content,
Calculate the bacterial strain degradation rate to organophosphorus pesticide；And by molybdenum blue colorimetric method, flat board face-off method measures rich phosphorus ability respectively and presses down
Plant pathogenic fungi ability processed, obtains bacterial strain MEW06.With MEW06 genome as template, use universal primer (27F and
1492R), amplification 16S rDNA sequence (see sequence table).MEW0616S rDNA PCR extension increasing sequence carries out same through NCBI Blast
Source comparison is analyzed, and finds that the sequence similarity of bacterial strain MEW06 and Serratia marcescens (GenBank:KT894728) is
99%, also it is 99% with the sequence similarity of Serratia nematodiphila (GenBank:KC172028).Use MEGA6
Carry out phylogenetic analysis understand, Serratia sp.MEW06 16S rDNA sequence evolve on and serratia marcescens
(Serratia marcescens) is close with addicted to nematicide Serratieae (Serratia nematodiphila).
Bacterial strain Serratia sp.MEW06 of the present invention is G-, amphimicrobian, in subsphaeroidal rod-short, does not produce spore, Zhousheng
Flagellum, without pod membrane.On LB flat board, cultivating 24h for 28 DEG C, bacterium colony is cerise, convex, smooth surface, moistening, opaque, limit
Edge is irregular, and size is 0.9-1.2 μ m 1.5-2.0 μm.During 28 DEG C of cultivations, bacterium solution presents cerise, and 37 DEG C do not show when cultivating
Color, result is shown in Fig. 1.Physiological biochemical property is: ODC Ornithine decarboxylase, esterase and DNA enzymatic are positive, and arabinose fermentation feminine gender.
Combining form, physiological and biochemical index and 16S rDNA sequence analysis (see sequence table), be initially identified as Serratia
Bacterium.
Case study on implementation 2
By the Serratia sp.MEW06 bacterial strain of purification, after activated overnight, regulate bacterium solution OD600=1.0, by 4% connect
The amount of kind is forwarded in the fermentation culture containing finite concentration content glyphosate or Rogor cultivate, and degrades in 1d, 3d, 5d sampling and measuring
Rate.Fermentative medium formula is as follows: peptone 2g/L, yeast extract 1g/L, NaCl 2g/L, pH7.0-7.2.Result shows,
Serratia sp.MEW06 is fabulous to the degradation effect of Rogor, when processing low concentration Rogor (50mg/L), cultivates 5d, supernatant
Being substantially not detectable Rogor concentration, degradation rate nearly reaches 100%.Process high concentration Rogor (200mg/L), cultivate 7d
After, measure degradation rate and reach 90.12%.MEW06 processes glyphosate ability and is weaker than Rogor, but under 50mg/L glyphosate concentration,
Cultivating 7d, degradation rate is about 95%.To sum up, for the Rogor of variable concentrations, the Serratieae of present embodiment all has preferably
Degradation effect, for low concentration glyphosate, the Serratieae of present embodiment has preferable degradation effect, and result is shown in Fig. 2.
Case study on implementation 3
Serratieae list colony inoculation activated overnight in LB culture medium in Example 1, LB culture medium prescription is as follows: egg
White peptone 10g/L, yeast extract 5g/L, NaCl 10g/L, pH7.0-7.2, the inoculum concentration by 2% is inoculated in phosphorus content respectively
(in terms of P) as 10mg/L, in the PAM culture medium of 20mg/L, in 0,12,24,48,72h sampling and measuring poly-phosphorus rate, PAM culture medium
Formula is: CH3COONa 5g/L, NH4Cl2g/L, KH2PO40.022～0.0878g/L, MgSO4·7H2O 0.5g/L, CaCl2
0.2g/L, deionized water 1000mL, pH7.0～7.2.Result is as shown in table 1, when in solution, phosphorus content is 10mg/L, through 24h
Enrichment, solution has been substantially not detectable content of inorganic phosphorus, rich phosphorus rate reaches 99.94%, and in solution, phosphorus content is 20mg/L,
Through the enrichment of 24h, rich phosphorus rate reaches 63.79%, and concrete outcome is shown in Table 1.The Serratieae of present embodiment can be the highest
Imitate poly-phosphorus, be expected to be applied to the process of the preparation of biologic phosphorus fertilizer and phosphorus-containing wastewater.
Table 1 Serratia sp.MEW06 poly-phosphorus rate (%) when PAM culture medium culturing changes over
Case study on implementation 4
Take the eugonic sharp knife Fusariumsp of a diameter of 5mm, Fructus Mali pumilae wheel stricture of vagina bacterium, Semen arachidis hypogaeae leaf mold, Rhizoctonia solani, Fructus Fragariae Ananssae
Grey mold and the pathogen of Botrytis cinerea truffle are inoculated in the central authorities of PDA flat board, cultivate 2d for 28 DEG C.Put in the position of distance mycelia edge 2cm
Put the filter paper (D=6mm) of sterilizing, bacterium solution and the PBS (blank) of 60 μ L are dropped on filter paper respectively.28 DEG C are continued training
After supporting 2 days, observe the test sample suppression situation to Fungal hyphal growth.From figure 3, it can be seen that Serratia sp.MEW06
Fermentation liquid can substantially suppress sharp knife Fusariumsp, Fructus Mali pumilae wheel stricture of vagina bacterium, Semen arachidis hypogaeae leaf mold and the growth of Botrytis cinerea mycelia, to Oryza sativa L.
The withered bacterium of stricture of vagina and the pathogen of Botrytis cinerea have certain inhibitory action.
<110>Hubei University
<120>one strains have rich phosphorus, degrading organic phosphor and the Serratieae of suppression plant pathogenic fungi
<160>1
<210>1
<211>1400
<212>DNA
<213>Serratieae (Serratia sp.)
<400>1
gtcgagcggt agcacagggg gagcttgctc cccgggtgac gagcggcgga cgggtgagta 60
atgtctggga aactgcctga tggaggggga taactactgg aaacggtagc taataccgca 120
taacgtcgca agaccaaaga gggggacctt cgggcctctt gccatcagat gtgcccagat 180
gggattagct agtaggtggg gtaatggctc acctaggcga cgatccctag ctggtctgag 240
aggatgacca gccacactgg aactgagaca cggtccagac tcctacggga ggcagcagtg 300
gggaatattg cacaatgggc gcaagcctga tgcagccatg ccgcgtgtgt gaagaaggcc 360
ttcgggttgt aaagcacttt cagcgaggag gaaggtggtg agcttaatac gttcatcaat 420
tgacgttact cgcagaagaa gcaccggcta actccgtgcc agcagccgcg gtaatacgga 480
gggtgcaagc gttaatcgga attactgggc gtaaagcgca cgcaggcggt ttgttaagtc 540
agatgtgaaa tccccgggct aacctgggaa ctgcatttga aactggcaag ctagagtctc 600
gtagaggggg gtagaattcc aggtgtagcg gtgaaatgcg tagagatctg gaggaatacc 660
ggtggcgaag gcggccccct ggacgaagac tgacgctcag gtgcgaaagc gtggggagca 720
aacaggatta gataccctgg tagtccacgc tgtaaacgat gtcgatttgg aggttgtgcc 780
cttgaggcgt ggcttccgga gctaacgcgt taaatcgacc gcctggggag tacggccgca 840
aggttaaaac tcaaatgaat tgacgggggc ccgcacaagc ggtggagcat gtggtttaat 900
tcgatgcaac gcgaagaacc ttacctactc ttgacatcca gagaactttc cagagatgga 960
ttggtgcctt cgggaactct gagacaggtg ctgcatggct gtcgtcagct cgtgttgtga 1020
aatgttgggt taagtcccgc aacgagcgca acccttatcc tttgttgcca gcggttcggc 1080
cgggaactca aaggagactg ccagtgataa actggaggaa ggtggggatg acgtcaagtc 1140
atcatggccc ttacgagtag ggctacacac gtgctacaat ggcgtataca aagagaagcg 1200
acctcgcgag agcaagcgga cctcataaag tacgtcgtag tccggattgg agtctgcaac 1260
tcgactccat gaagtcggaa tcgctagtaa tcgtagatca gaatgctacg gtgaatacgt 1320
tcccgggcct tgtacacacc gcccgtcaca ccatgggagt gggttgcaaa agaagtaggt 1380
agcttaacct tcgggagggc 1400
Claims (3)
1. a strain has rich phosphorus, degrading organic phosphor and the Serratieae of suppression plant pathogenic fungi, it is characterised in that: this sand Lei Shi
Bacterium is SerratiaSerratiaSp. MEW06, December in 2015 is preserved in China typical culture collection center on the 25th and protects
Hiding, deposit number is CCTCC NO. M 2015779.
A strain the most as claimed in claim 1 has rich phosphorus, degrading organic phosphor and the Serratieae of suppression plant pathogenic fungi,
It is characterized in that: this bacterial strainSerratiaSp. MEW06 is used for being enriched with Phos and/or degrading organic phosphor and/or preventing and treating plant
Pathogenic fungi.
A strain the most as claimed in claim 2 has rich phosphorus, degrading organic phosphor and the Serratieae of suppression plant pathogenic fungi,
It is characterized in that: this bacterial strainSerratiaSp. MEW06 degradable Rogor, glyphosate.
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| CN114908008A (en) * | 2022-05-07 | 2022-08-16 | 龙江环保集团股份有限公司 | Serratia marcescens strain and application thereof |
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| CN103122330A (en) * | 2012-12-31 | 2013-05-29 | 陈秀蓉 | Alpine grassland pasture endogenous serratia plymuthica strain GH010 and application thereof as well as microbial agent prepared from same and preparation method |
| CN104087544A (en) * | 2014-06-19 | 2014-10-08 | 徐州工程学院 | Engineering bacterium capable of degrading organophosphorus pesticides, and construction method and application thereof |
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| CN102676439A (en) * | 2012-06-26 | 2012-09-19 | 四川大学 | Serratia strain and method for disposing methidathion pesticide residues on surfaces of soil and fruits/vegetables |
| CN103122330A (en) * | 2012-12-31 | 2013-05-29 | 陈秀蓉 | Alpine grassland pasture endogenous serratia plymuthica strain GH010 and application thereof as well as microbial agent prepared from same and preparation method |
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