CN107217016A - One plant has the endophytic Bacillus bacterial strain ZY122 for suppressing rice sheath blight disease and its application - Google Patents
One plant has the endophytic Bacillus bacterial strain ZY122 for suppressing rice sheath blight disease and its application Download PDFInfo
- Publication number
- CN107217016A CN107217016A CN201710309388.7A CN201710309388A CN107217016A CN 107217016 A CN107217016 A CN 107217016A CN 201710309388 A CN201710309388 A CN 201710309388A CN 107217016 A CN107217016 A CN 107217016A
- Authority
- CN
- China
- Prior art keywords
- bacillus
- bacterial strain
- sheath blight
- blight disease
- rice sheath
- 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.)
- Granted
Links
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
- C12R2001/07—Bacillus
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/10—Animals; Substances produced thereby or obtained therefrom
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Virology (AREA)
- Environmental Sciences (AREA)
- Pest Control & Pesticides (AREA)
- Dentistry (AREA)
- Plant Pathology (AREA)
- Agronomy & Crop Science (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biomedical Technology (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
There is the endophytic Bacillus bacterial strain ZY122 (Bacillus vanilleaZY122) for suppressing rice sheath blight disease and its application the present invention relates to one plant, the bacterial strain was preserved in Guangdong Province's Culture Collection (GDMCC) on April 11st, 2017, and deposit number is GDMCCNo.60161.The bacillus vanilleaZY122 that the present invention is provided can be used for the biological control of rice sheath blight disease, to the pathogen thanatephorus cucumeris(frank) donk Thanatephoruscucumeris of rice sheath blight disease bacteriostasis rate up to 81.9%, prevention effect is up to more than 70.0%.The present invention carries out separation screening to the strain of Antagonistic Endophytic present in healthy plant paddy rice, is that the biological control of rice sheath blight disease lays the foundation using rice sheath blight disease pathogen as controlling object.The reasonable application for the Bacillus strain ZY122 that the present invention is provided can reduce the environmental pollution that chemical agent is caused; protecting ecology is conducive to balance; to the artificial cultivation of paddy rice and increase income volume increase and can play very big raising and facilitation, with certain Research Significance.
Description
Technical field
The present invention relates to technical field of plant disease biological control, in particular it relates to which one plant has and suppresses rice sheath blight disease
Endophytic Bacillus bacterial strain ZY122 and its application.
Background technology
Paddy rice, as one of big staple food grain in the world three, is the staple food grain of global about 50% population.Its main integrated distribution the torrid zone or
Semi-tropical areas of high temperature and rainfall, such as China Yangtze river basin, the Delta of the Pearl River and the Sichuan Basin, South East Asia Mainland etc., wherein nearly 90%
Originate in Asia.About 155,000,000 hectares of current world Monitoring of Paddy Rice Plant Area, China's Rice Cropping face is 31,000,000 hectares, accounts for generation
The 20% of boundary's cultivated area；The paddy rice total output of China ranks first in the world, and per unit area yield occupies the world the 5th.Paddy rice is mainly distributed in China
In six big producing regions, it is respectively：South China Double Cropping rice makees band；East China, the single double rice cropping band in Central China；Northwest drying of rice makees band；North China is single
Season, rice made band；Northeast early rice makees band and southwest plateau rice makees band.As population constantly increases, demand of the China to paddy
Also it is increasing, how to ensure that the development balanced to China's rice industry of eating rice production and consumption proposes severe challenge.
Under the influence of the factors such as China's freshwater resources anxiety, labor shortage and production cost increase, Rice Cropping increases grain with increasing income
Contradiction, along with global climate and ecological environment go from bad to worse, causes the various pest and disease damages of paddy rice to be presented increased
Situation.
Rice sheath blight disease is one of Major Diseases in current Rice Production, and the disease is by thanatephorus cucumeris(frank) donkThanatephoruscucumerisIt is caused, thanatephorus cucumeris(frank) donkThanatephoruscucumerisBelong to Basidiomycotina fungi,
Occurrence scope is wide, harm is serious, preventing and treating is difficult.The disease prevents paddy rice from earing, or the not plump paddy of heading is more, and grain declines again.It is right
In the existing traditional chemical agent of preventing and treating of rice sheath blight disease, also there is emerging biological Control Technology.The country is related to paddy rice line
The patented technology of rot there are about 400 remainders, and the wherein related patent of chemical agent accounts for 60%；Biotechnology accounts for 30%；Chemistry
The patented technology combined with both biological controls accounts for 10%.
Bacillus is universally present in nature, with the characteristics of breeding is quick, metabolism is fast；The important spy of the category bacterium
Property is can to produce the gemma to unfavorable conditions with special resistance.They exist in interior soil, plant tissue, water and sky
It is closely related with the conversion of nature material, soil fertility, environmental sanitation in gas.They are strong to organic matter decomposition power simultaneously, increase
While growing, the amylase of high activity can be disengaged, the materials such as a variety of organic acids, enzyme, physiologically active, and other a variety of appearances is synthesized
The nutrient being easily utilized, promotes plant growth.With the two packing spaces sexual involution and alkalization of soils of soil, bacillus can be
Survive and grow in adverse circumstance, the diversity of various enzymes and increase edaphon is provided for soil.Endophyte of plant is certain
Fungi or bacterium that stage or whole stages are moved in inside the tissue and organ of health plant.It is prevalent in higher plant
In, woody, herbaceous plant has endogenetic bacteria in monocotyledon and dicotyledon.Endophyte can not only be carried to plant
For needed nutrient matter and some hormones, to the growing of plant, resist disease pest invasion and attack and poor environment etc. have it is extensive
Biological action, can also strengthen Genes For Plant Tolerance adverse circumstance, the ability of disease and insect resistance, turn into potential micro- in biological control at present
Biological pesticide, yield increasing fungus are used as potential biological and ecological methods to prevent plant disease, pests, and erosion carrier bacterium.
Endophyte is present in plant, can colonize and conduct for a long time in plant, be difficult by external environmental condition
Influence, the compound that the bioactive substance separated from endophyte of plant for having studies have shown that 51% is not found before being, this
The exploitation for the frontier originated to antibacterial substance is by with very great economic implications.Screening can have short of money to pathogen
The bacillus of anti-effect has important in terms of the biological control, promotion plant growth, improvement ecological environment of soil of disease
Meaning.
The content of the invention
Have it is an object of the invention to overcome the deficiencies of the prior art and provide one plant and suppress to sprout in rice sheath blight disease
Spore bacillus strain ZY122, the endophytic Bacillus bacterial strain ZY122 that the present invention is provided can be used for suppressing rice sheath blight disease, and it is to water
The pathogen thanatephorus cucumeris(frank) donk of the sheath and culm blight of riceThanatephoruscucumerisBacteriostasis rate up to 81.9%, prevention effect is reachable
More than 70.0%.
Another object of the present invention is to provide above-mentioned endophytic Bacillus bacterial strain ZY122 in the biology of rice sheath blight disease to prevent
Application in controlling.
Another object of the present invention is to provide endophytic Bacillus bacterial strain ZY122 to prepare the life for suppressing rice sheath blight disease
Application in thing preparation.
It is a kind of with the biological agent for suppressing rice sheath blight disease another object of the present invention is to provide.
To achieve the above object, the present invention is adopted the following technical scheme that：
One plant has the endophytic Bacillus bacterial strain ZY122 (Bacillus vanilleaZY122) for suppressing rice sheath blight disease, institute
State bacterial strain and be preserved in Guangdong Province's Culture Collection (GDMCC) on April 11st, 2017, preservation address is Guangzhou
Martyr Road 100, deposit number is GDMCCNo.60161.
Heretofore described bacterial strain contains the 16SrDNA gene orders as shown in SEQ1.
The bacillus of the present inventionBacillus vanilleaZY122 colony morphology characteristics：The bacterial strain is on LB culture mediums
The speed of growth is very fast（37 DEG C, 20h）, it is mucus type bacterial strain in greyish white shape, cell is sub-translucent, is in（Long 1.3~1.8um is wide by 0.4
~0.7um）It is shaft-like, there is projection in the middle of bacterium colony, gemma, peritrichous can be produced.
The bacillus of the present inventionBacillus vanilleaZY122 physio-biochemical characteristics：
Gram-positive bacteria, amphimicrobian, pH growth scopes pH4.0~8.0（Most preferably 5.0~7.0）, resistance to NaCl can 0~
It can be grown under the conditions of 8% (W/V)（Most preferably 1~4%）, it is 20~45 DEG C to be suitable for growth temperature interval（Most preferably 28~7 DEG C）,
DNA G+C contents are 46.4mol%.It is the positive to produce ammonia test, acetyl formaldehyde Methanol determination；Urinate enzyme test, methyl red determine,
Catalase test, citrate utilization test are feminine gender.Gentiobiose, α-D-lactose, β-methyl-D-grape can efficiently be utilized
Glucosides, glycogen, D- xyloses then grow slow as carbon source using arabinose, melibiose, trehalose and turanose as carbon source
Slowly.Bacterial strain is respectively provided with tolerance to 50 μ g/mL ampicillins, 100 μ g/mL streptomysins, 300 μ g/mL tetracyclines；And to OK a karaoke club
Mycin, gentamicin, chloramphenicol, neomycin, erythromycin poor resistance, low concentration（5μg/mL）This several antibiotic just can press down
Make its growth.
The bacillus that the present invention is providedBacillus vanilleaZY122 is through 16SrDNA sequence alignments
(EZBiocloud databases) withBacillusvanilleaThe homology of XY18 (T) bacillus is 99.42%；The bacterial strain
16SrDNA sequences are as shown in sequence table.
Bacillus of the present inventionBacillus vanilleaThe determination of ZY122 molecular classification status：
Expanded using bacteria 16 S rRNA genes universal primer 25f and 1492r, PCR primer is directly subjected to sequencing.
The DNA sequence dna input EZBiocloud of acquisition is compared, genus and species of the bacterial strain of the present invention in taxology are primarily determined that
Position.As a result find that the bacillus of the present invention belongs to bacillusBacillus, withBacillusvanilleaXY18(T)
The homology of bacillus is 99.42%, but the sequence of the two is not quite identical, the difference for having 0.58%, and it is of the same race to show the two
Interior different bacterial strain.With reference to above-mentioned physio-biochemical characteristics, 16SrDNA sequence analyses, bacillus of the inventionBacillus vanilleaZY122 should belong to bacillusBacillus。
The bacillus of the present inventionBacillus vanilleaZY122 can be used for the biological control of rice sheath blight disease.More
Body, the Bacillus strain ZY122 can be applied to suppress rice sheath blight disease pathogen thanatephorus cucumeris(frank) donkThanatephoruscucumeris.BacillusBacillus vanilleaZY122 dies to the pathogen melon of rice sheath blight disease
Lead fungiThanatephoruscucumerisBacteriostasis rate is up to 81.9%, to the pathogen thanatephorus cucumeris(frank) donk of rice sheath blight diseaseThanatephoruscucumerisPrevention effect is up to more than 70.0%.
Applications of the above-mentioned endophytic Bacillus bacterial strain ZY122 in the biological agent for suppressing rice sheath blight disease is prepared is also at this
Within the protection domain of invention.
It is a kind of to have in the biological agent for suppressing rice sheath blight disease, the biological agent containing interior described in claim 1
Raw Bacillus strain ZY122.
Compared with prior art, the present invention has the advantages that：
The present invention is by using selective medium, and the pathogen thanatephorus cucumeris(frank) donk using rice sheath blight disease is controlling object, to growth
The Antagonistic Endophytic present in long-term unmanured healthy wild rice plants is separated, purified, and obtains one plant of antagonism water
The bacillus of the sheath and culm blight of rice, is that the biological control of rice sheath blight disease is laid a good foundation.The endophytic Bacillus that the present invention is provided
Bacterial strain ZY122 reasonable application can reduce chemical agent pollution on the environment, be conducive to protecting ecology to balance, to paddy rice
Cultivation and increase income volume increase and can play very big raising and facilitation, with certain Research Significance.
Brief description of the drawings
Fig. 1 is the bacillus that embodiment 1 is providedBacillus vanilleaZY122 dilution spreads on LB culture mediums
Bacterium colony picture；
Fig. 2 is the pathogen thanatephorus cucumeris(frank) donk of rice sheath blight diseaseThanatephoruscucumerisControl group is in PDA culture medium
Growth figure；
Fig. 3 is the bacillus that embodiment 1 is providedBacillus vanilleaZY122 is to pathogen thanatephorus cucumeris(frank) donkThanatephoruscucumerisAntagonistic effect figure；
Fig. 4 is the bacillus that embodiment 1 is providedBacillus vanilleaThe PCR of ZY122 complete genome DNAs and 16SrDNA
The agarose gel electrophoresis figure of product 1%.
Embodiment
Technical scheme is further described with reference to chart, but this explanation does not do any form side
The restriction in face, it is every technical solution of the present invention to be modified or equivalent substitution, without departing from the essence of technical solution of the present invention
God and scope, all should cover in protection scope of the present invention.
The bacillus vanilleaZY122 of embodiment 1 separation, screening and chemotaxonomy status
Determine that the biocontrol bacterial strain bacillus vanilleaZY122 for the suppression rice sheath blight disease that this implementation is provided is preserved in
Guangdong Province's Culture Collection (GDMCC), preservation address Xianlie Middle Road, Guangzhou City 100, preservation date is 2017
April 11, deposit number is GDMCCNo.60161.
The bacillus of present embodimentBacillusZY122 derives from the wild rice field in Guangxi Wuzhou, using selective training
Support base and endogenetic bacteria separation acquisition is carried out to wild rice healthy plant, specific method is as follows：
（1）The separation of wild rice endophyte
The wild rice material of collection is cleaned with distilled water, appropriate root, stem, leaf texture is chosen, fragment is cut into sterile scissors
Shape, is put into sterile petri dish, soaks 3min with 75% ethanol, is used after then soaking 3min, taking-up in 2%NaClO solution
Aseptic water washing 3 times, each 5min, the sterilized water coating LB of last time cleaning, as control test, whether materials disinfection is thorough
Bottom.Sterilized water is added in sterile mortar and grinds to form homogenate, after ten times of homogenate dilution, 100uL is drawn respectively with liquid-transfering gun
On to sterile LB solid plates, 37 DEG C of constant incubator cultures are put into after smearing uniformly, the growing state of thalline are observed, with inoculation
Ring picking growing way is good and the different lawn of form, and Secondary Culture repeatedly is carried out with plate streak, until color, the shape of bacterium colony
Shape, size, quality are as transparency.Finally by simple dyeing（Carbolic acid azaleine is dyed）And sediments microscope inspection（Oil mirror）,
Its form is further looked at, so that length is homogeneous, width is consistent, staining conditions are unified for bacterial strain purification standard.Bacterial strain after purification
- 20 DEG C and -80 DEG C preserve respectively in 15% glycerine.
（2）The primary dcreening operation of endophyte antagonism rice banded sclerotial blight pathogen
Method is stood facing each other to wild rice endophyte antagonism banded sclerotial blight disease fungus progress primary dcreening operation using flat board.By rice sheath blight disease pathogen
It is 10 that concentration is made of sterilized water8/mL-1Bacteria suspension, take 20uL flat boards to be coated in PDA culture medium, then in the PDA of coating
Mark signature line separates 4 regions on flat board, in each area the heart accessed respectively with sterile toothpick it is a small amount of be purified into it is wild
Rice endophyte, is put in 28 DEG C of constant incubators and cultivates, the presence or absence of observed and recorded inhibition zone and size, are repeated 3 times altogether after 5 days.
Select 5 plants of maximum endophytes of inhibition zone width and carry out secondary screening.
（3）The secondary screening of endophyte antagonism rice banded sclerotial blight pathogen
Method is stood facing each other to endophyte antagonizing pathogenic fungi progress secondary screening using flat board.Rice sheath blight disease pathogen is made of sterilized water
Concentration is 108/mL-1Bacteria suspension, then take 20uL to be coated with PDA culture medium, 4 separated having been coated with mark signature line on flat board
Individual region, each regional center accesses the endophyte that preliminary screening comes out antagonism with sterile toothpick respectively, if sterilized water
For control, it is repeated 3 times.It is placed under 28 DEG C of constant temperatures and cultivates, fungistatic effect is observed after 5 days, measures inhibition zone size.Select effect
One plant of really best bacterium, saves backup.
Bacteriostasis rate (%)=(treatment group antibacterial circle diameter-control group antibacterial circle diameter/treatment group antibacterial circle diameter) ×
100%
Culture medium prescription applied in the present embodiment is as follows：
LB culture mediums：Yeast extract 5g, bacteriological peptone 10g, Nacl5g, Agar20g, sterilized water 1L, PH=7.0.
PDA culture medium：Peeled potatoes 200g, glucose 20g, Agar20g, sterilized water 1L, natural PH.
The bacillus that Fig. 1 provides for the present embodimentBacillus vanilleaZY122 dilution spreads on LB culture mediums
Bacterium colony picture, left figure be the bacterium colony back side shine, right figure be bacterium colony front shine.The bacillus of the present invention can be observed by Fig. 1Bacillus vanilleaZY122 colony morphology characteristics：Bacterial strain speed of growth on LB culture mediums is very fast（37 DEG C, 20h）,
It is mucus type bacterial strain in greyish white shape, cell is sub-translucent, is in（Wide 0.4~the 0.7um of long 1.3~1.8um）It is shaft-like, have in the middle of bacterium colony
Projection, can produce gemma, peritrichous.
Fig. 2 is the pathogen thanatephorus cucumeris(frank) donk of rice sheath blight diseaseThanatephoruscucumerisControl group is cultivated in PDA
Growth figure on base, left figure is that thanatephorus cucumeris(frank) donk front is shone, and right figure is shone for the thanatephorus cucumeris(frank) donk back side；Fig. 3 is bacillus
VanilleaZY122 to the pathogen thanatephorus cucumeris(frank) donk Thanatephoruscucumeris of rice sheath blight disease fungistatic effect figure,
Left figure is shone for antibacterial front, and right figure is shone for the antibacterial back side.BacillusBacillusPathogen melons of the ZY122 to rice sheath blight disease
Die lead fungiThanatephoruscucumerisBacteriostasis rate up to 81.9%.
（4）BacillusBacillus vanilleaThe determination of ZY122 chemotaxonomy status
Picking antagonistic BacillusBacillus vanilleaZY122 is inoculated in LB solid mediums, and thalline is collected after 12h
And strain gene group DNA is extracted, with the size of 1% agarose electrophoresis validating DNA pack section.The gemma that Fig. 4 provides for the present embodiment
BacillusBacillus vanilleaThe agarose gel electrophoresis figure of ZY122 complete genome DNAs and 16SrDNA PCR primer 1%,
Wherein left figure is 1% agarose gel electrophoresis figure of complete genome DNA, and right figure is coagulated for the agarose of 16Sr DNA PCR primer 1%
Gel electrophoresis figure.As shown in Figure 4：The electrophoretic band of full-length genome is single clear in left figure, and full-length genome is more than 10000bp；Right figure
In PCR primer electrophoretic band it is single clear, fragment length is about 1500bp or so.
Using genomic DNA as template, using bacterial 16 S rDNA gene universal primers：- the AGAGTTTGATC- of sense primer 5 '
CTGGCTCAG-3 ' and-the AAGGAGGTGATCCAGCC-3 of anti-sense primer 5 '；
PCR reaction systems：10 × PCRbuffer2.5 μ L, dNTP2.0 μ L, each 0.5 μ L of upstream and downstream primer, the μ L of DNA profiling 1.0,
The μ L of Taq DNA polymerase 0.2, mend distilled water to 25 μ L.PCR reaction conditions：94 DEG C of pre-degeneration 5min；94 DEG C are denatured 45s, 55 DEG C
Anneal 45s, 72 DEG C of extension 1.5min, cyclic amplification 30 times：Last 72 DEG C of extensions 10min.With the checking PCR productions of 1% agarose electrophoresis
The size of thing DNA fragmentation, as shown in Figure 4, bacillusBacillus vanilleaZY122 DNA PCR primers pass through gel
Single band is arrived in electrophoresis amplification, and band is clear, and size is about 1500bp.Beijing is entrusted to hold up section's new industry biotechnology PCR primer
Co., Ltd is sequenced, and the DNA sequence dna input GenBank of acquisition carries out Blast comparisons, primarily determines that the gemma bar of the present invention
BacteriumBacillus vanilleaThe position of genus and species of the ZY122 bacterial strains in taxology.As a result described bacillus is foundBacillus vanilleaZY122, withBacillusvanilleaThe homology of XY18 (T) bacillus is 99.42%, with reference to
Above-mentioned morphological feature, physio-biochemical characteristics, 16SrDNA Phylogenetic Analysis and Literature Consult, bacillus of the inventionBacillus vanilleaZY122 should belong to bacillusBacillus。
Sequencing result is as follows：
GCTATAATGCAGTCGAGCGGACAGATGGGAGCTTGCTCCCTGATGTTAGCGGCGGACGGGTGAGTAACACGTG
GGTAACCTGCCTGTAAGACTGGGATAACTCCGGGAAACCGGGGCTAATACCGGATGGTTGTCTGAACCGCATGGTTC
AGACATAAAAGGTGGCTTCGGCTACCACTTACAGATGGACCCGCGGCGCATTAGCTAGTTGGTGAGGTAACGGCTCA
CCAAGGCGACGATGCGTAGCCGACCTGAGAGGGTGATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGG
GAGGCAGCAGTAGGGAATCTTCCGCAATGGACGAAAGTCTGACGGAGCAACGCCGCGTGAGTGATGAAGGTTTTCGG
ATCGTAAAGCTCTGTTGTTAGGGAAGAACAAGTGCCGTTCAAATAGGGCGGCACCTTGACGGTACCTAACCAGAAAG
CCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGAATTATTGGGCGTAAAGGG
CTCGCAGGCGGTTTCTTAAGTCTGATGTGAAAGCCCCCGGCTCAACCGGGGAGGGTCATTGGAAACTGGGGAACTTG
AGTGCAGAAGAGGAGAGTGGAATTCCACGTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGG
CGACTCTCTGGTCTGTAACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCAC
GCCGTAAACGATGAGTGCTAAGTGTTAGGGGGTTTCCGCCCCTTAGTGCTGCAGCTAACGCATTAAGCACTCCGCCT
GGGGAGTACGGTCGCAAGACTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATT
CGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCTCTGACAATCCTAGAGATAGGACGTCCCCTTCGGGGGCA
GAGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCT
TGATCTTAGTTGCCAGCATTCAGTTGGGCACTCTAAGGTGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACG
TCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGACAGAACAAAGGGCAGCGAAACCGCGAG
GTTAAGCCAATCCCACAAATCTGTTCTCAGTTCGGATCGCAGTCTGCAACTCGACTGCGTGAAGCTGGAATCGCTAG
TAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAGAGTTTGT
AACACCCGAAGTCGGTGAGGTAACCTTTAGGAGCCAGCCGCCGAAG
Pathogen in the present embodiment is from the isolated thanatephorus cucumeris(frank) donk of rice stem base portionThanatephoruscucumeris。
Applications of the bacillus vanilleaZY122 of embodiment 2 in the biological control of rice sheath blight disease
Bacillus in present embodimentBacillus vanilleaZY122 can be used for the biological control of rice sheath blight disease, preventing and treating
Effect experiment is as follows：
Set three experimental groups to carry out potted plant experiments, respectively healthy control group, susceptible control group and prevent and treat group, every group of 20 plants of water
Rice.Use thanatephorus cucumeris(frank) donkThanatephoruscucumerisIt is 1 × 10 to be configured to concentration8Individual/mL spore suspension.By paddy rice
Carry out after surface sterilization, 20 μ L sterilized waters, 20 μ of susceptible control group and each injection of preventing and treating group are injected to healthy control group with needle point method
It is inoculated in after L thanatephorus cucumeris(frank) donk suspension in flowerpot.And preventing and treating group plant root pouring 3mL antagonistic Bacillus nutrient solutions is (dense
Spend for 0.6 × 107~1 × 109cfu/L).Ensure that the paddy rice of each experimental group grows at shady and cool ventilation, poured daily,
Observed after 15d and record rice pathogenesis situation.
Prevention effect=(susceptible group of plant number-control group disease plant number-preventing and treating group disease plant number)/inoculation plant number
× 100%.
The bacillus of table 1Bacillus vanilleaZY122 suppresses thanatephorus cucumeris(frank) donkThanatephoruscucumeris
Test
Understood according to experimental result, bacillusBacillus vanilleaZY122 is to thanatephorus cucumeris(frank) donkThanatephoruscucumerisPrevention effect up to more than 70.0%.
SEQUENCE LISTING
<110>Agricultural University Of South China
<120>One plant has the endophytic Bacillus bacterial strain ZY122 for suppressing rice sheath blight disease and its application
<130> 1
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1428
<212> DNA
<213>Bacillus vanilleaZY122 16srDNA
<400> 1
gctataatgc agtcgagcgg acagatggga gcttgctccc tgatgttagc ggcggacggg 60
tgagtaacac gtgggtaacc tgcctgtaag actgggataa ctccgggaaa ccggggctaa 120
taccggatgg ttgtctgaac cgcatggttc agacataaaa ggtggcttcg gctaccactt 180
acagatggac ccgcggcgca ttagctagtt ggtgaggtaa cggctcacca aggcgacgat 240
gcgtagccga cctgagaggg tgatcggcca cactgggact gagacacggc ccagactcct 300
acgggaggca gcagtaggga atcttccgca atggacgaaa gtctgacgga gcaacgccgc 360
gtgagtgatg aaggttttcg gatcgtaaag ctctgttgtt agggaagaac aagtgccgtt 420
caaatagggc ggcaccttga cggtacctaa ccagaaagcc acggctaact acgtgccagc 480
agccgcggta atacgtaggt ggcaagcgtt gtccggaatt attgggcgta aagggctcgc 540
aggcggtttc ttaagtctga tgtgaaagcc cccggctcaa ccggggaggg tcattggaaa 600
ctggggaact tgagtgcaga agaggagagt ggaattccac gtgtagcggt gaaatgcgta 660
gagatgtgga ggaacaccag tggcgaaggc gactctctgg tctgtaactg acgctgagga 720
gcgaaagcgt ggggagcgaa caggattaga taccctggta gtccacgccg taaacgatga 780
gtgctaagtg ttagggggtt tccgcccctt agtgctgcag ctaacgcatt aagcactccg 840
cctggggagt acggtcgcaa gactgaaact caaaggaatt gacgggggcc cgcacaagcg 900
gtggagcatg tggtttaatt cgaagcaacg cgaagaacct taccaggtct tgacatcctc 960
tgacaatcct agagatagga cgtccccttc gggggcagag tgacaggtgg tgcatggttg 1020
tcgtcagctc gtgtcgtgag atgttgggtt aagtcccgca acgagcgcaa cccttgatct 1080
tagttgccag cattcagttg ggcactctaa ggtgactgcc ggtgacaaac cggaggaagg 1140
tggggatgac gtcaaatcat catgcccctt atgacctggg ctacacacgt gctacaatgg 1200
acagaacaaa gggcagcgaa accgcgaggt taagccaatc ccacaaatct gttctcagtt 1260
cggatcgcag tctgcaactc gactgcgtga agctggaatc gctagtaatc gcggatcagc 1320
atgccgcggt gaatacgttc ccgggccttg tacacaccgc ccgtcacacc acgagagttt 1380
gtaacacccg aagtcggtga ggtaaccttt aggagccagc cgccgaag 1428
Claims (8)
1. one plant has the endophytic Bacillus bacterial strain ZY122 (Bacillus vanilleaZY122) for suppressing rice sheath blight disease,
Characterized in that, the bacterial strain was preserved in Guangdong Province's Culture Collection (GDMCC), preservation on April 11st, 2017
Numbering is GDMCC No.60161.
2. endophytic Bacillus bacterial strain ZY122 according to claim 1, it is characterised in that the bacterial strain contains such as SEQ1 institutes
The 16SrDNA gene orders shown.
3. endophytic Bacillus bacterial strain ZY122 according to claim 1, it is characterised in that the bacterial strain is Gram-positive
Bacterium, amphimicrobian, are in greyish white shape on LB culture mediums, are mucus type bacterial strain, cell is sub-translucent, in a length of 1.3~1.8 μm, width
It is shaft-like for 0.4~0.7 μm；There is projection in the middle of bacterium colony, gemma, peritrichous can be produced.
4. endophytic Bacillus bacterial strain ZY122 according to claim 1, it is characterised in that the bacterial strain with gentiobiose, α-
One or more in D-lactose, β-methyl-D-glucoside, glycogen or D- xyloses are carbon source.
5. applications of the endophytic Bacillus bacterial strain ZY122 in the biological control of rice sheath blight disease described in claim 1.
6. apply according to claim 5, it is characterised in that the application is that Bacillus strain ZY122 is suppressing paddy rice
Application in banded sclerotial blight pathogen thanatephorus cucumeris(frank) donk Thanatephoruscucumeris.
7. the answering in the biological agent for suppressing rice sheath blight disease is prepared of endophytic Bacillus bacterial strain ZY122 described in claim 1
With.
8. a kind of have the biological agent for suppressing rice sheath blight disease, it is characterised in that is wanted in the biological agent containing having the right
Seek endophytic Bacillus bacterial strain ZY122 described in 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710309388.7A CN107217016B (en) | 2017-05-04 | 2017-05-04 | One plant has the endophytic Bacillus bacterial strain ZY122 for inhibiting rice sheath blight disease and its application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710309388.7A CN107217016B (en) | 2017-05-04 | 2017-05-04 | One plant has the endophytic Bacillus bacterial strain ZY122 for inhibiting rice sheath blight disease and its application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107217016A true CN107217016A (en) | 2017-09-29 |
| CN107217016B CN107217016B (en) | 2019-10-29 |
Family
ID=59944660
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710309388.7A Active CN107217016B (en) | 2017-05-04 | 2017-05-04 | One plant has the endophytic Bacillus bacterial strain ZY122 for inhibiting rice sheath blight disease and its application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107217016B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110628681A (en) * | 2019-10-11 | 2019-12-31 | 沈阳农业大学 | Bacillus altitudinis strain and application thereof in rice sheath blight |
| CN110878268A (en) * | 2019-11-26 | 2020-03-13 | 湖南新九方科技有限公司 | Bacillus megaterium for resisting rice sheath blight and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105695366A (en) * | 2016-04-05 | 2016-06-22 | 中国农业科学院植物保护研究所 | Bacillus strain and application thereof |
| CN105907683A (en) * | 2016-06-02 | 2016-08-31 | 四川省农业科学院植物保护研究所 | Bacillus amyloliquefaciens cd5bc and application thereof |
-
2017
- 2017-05-04 CN CN201710309388.7A patent/CN107217016B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105695366A (en) * | 2016-04-05 | 2016-06-22 | 中国农业科学院植物保护研究所 | Bacillus strain and application thereof |
| CN105907683A (en) * | 2016-06-02 | 2016-08-31 | 四川省农业科学院植物保护研究所 | Bacillus amyloliquefaciens cd5bc and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| 农倩: "水稻内生细菌对水稻纹枯病的防治作用研究", 《万方学位论文数据库》 * |
| 黎起秦等: "广西水稻内生细菌的动态分布及其对水稻纹枯病菌的拮抗作用", 《中国生物防治》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110628681A (en) * | 2019-10-11 | 2019-12-31 | 沈阳农业大学 | Bacillus altitudinis strain and application thereof in rice sheath blight |
| CN110628681B (en) * | 2019-10-11 | 2021-04-30 | 沈阳农业大学 | Bacillus altitudinis strain and application thereof in rice sheath blight |
| CN110878268A (en) * | 2019-11-26 | 2020-03-13 | 湖南新九方科技有限公司 | Bacillus megaterium for resisting rice sheath blight and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107217016B (en) | 2019-10-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106754557B (en) | Bacillus subtilis YBM-4 and application thereof in preventing and treating tobacco black shank and promoting growth | |
| CN107779420B (en) | Two-strain endogenous Bacillus belgii for antagonizing tobacco bacterial wilt and application thereof | |
| CN112899171B (en) | Rapex leucocytochys LL210, application thereof and biocontrol microbial inoculum | |
| CN106434409B (en) | The rice endogeny rayungus OsiLf-2 of one plant of external efficiently antagonism Pyricularia oryzae | |
| CN102220246B (en) | Blueberry mycorrhizal fungi (coprinus micaceus) and separation method and application thereof | |
| CN106148232B (en) | The bacterium bacterial strain and its application of one plant of antagonism plant pathogenetic bacteria | |
| CN107136122B (en) | Biocontrol microbial inoculum for preventing and treating potato late blight | |
| CN110669691B (en) | Bacillus megaterium for preventing and treating plant nematode diseases and application thereof | |
| CN113717901A (en) | Bacillus belgii and application thereof in prevention and treatment of various vegetable diseases | |
| CN108641989B (en) | Bacillus methylotrophicus and application thereof | |
| CN103109870B (en) | Application of fermentation supernatant liquid of pseudomonas | |
| CN113846039A (en) | Bacillus belgii and application thereof | |
| CN114381379B (en) | Adhesive film fungus strain TP-8 with dendrobium seedling tillering improving capability and application thereof | |
| CN110172423B (en) | Bacillus belgii and application thereof in preventing and controlling root-knot nematodes | |
| CN109749953B (en) | Bacillus cereus, microbial inoculum and preparation method and application thereof | |
| CN103981101B (en) | A kind of DSE bacterial strain and the application in sugarcane production thereof | |
| CN110317747A (en) | A kind of bacillus amyloliquefaciens JT68 and its application in prevention and treatment tea anthracnose | |
| CN107217016B (en) | One plant has the endophytic Bacillus bacterial strain ZY122 for inhibiting rice sheath blight disease and its application | |
| CN110172424B (en) | Bacillus methylotrophicus and application thereof in preventing and controlling root-knot nematodes | |
| CN114395486B (en) | Adhesive film fungus strain TP-3 with high growth promoting capability of dendrobium and application thereof | |
| CN114480171B (en) | Rhizobium sojae strain and application thereof | |
| CN104126508A (en) | Rapid mycorrhization method of orchid aseptic seedlings | |
| CN105420167B (en) | A kind of Bacillus cercus and its application | |
| CN114507618A (en) | Turkey mycorrhiza strain TP-11 with capacity of promoting growth of new leaves of dendrobium and application thereof | |
| CN106167767A (en) | The endogenetic fungus L 14 of preventing and treating banana blight and application 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 |