CN107217016B - One plant has the endophytic Bacillus bacterial strain ZY122 for inhibiting rice sheath blight disease and its application - Google Patents

One plant has the endophytic Bacillus bacterial strain ZY122 for inhibiting rice sheath blight disease and its application Download PDF

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CN107217016B
CN107217016B CN201710309388.7A CN201710309388A CN107217016B CN 107217016 B CN107217016 B CN 107217016B CN 201710309388 A CN201710309388 A CN 201710309388A CN 107217016 B CN107217016 B CN 107217016B
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bacillus
sheath blight
rice
blight disease
rice sheath
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CN107217016A (en
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谭志远
黄淑芬
郜晨
谭习羽
彭桂香
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South China Agricultural University
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South China Agricultural University
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    • 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
    • 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
    • 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

Abstract

The present invention relates to one plant to have the endophytic Bacillus bacterial strain ZY122 (Bacillus vanilleaZY122) for inhibiting rice sheath blight disease and its application, the bacterial strain is preserved in Guangdong Province's Culture Collection (GDMCC), deposit number GDMCCNo.60161 on April 11st, 2017.Bacillus vanilleaZY122 provided by the invention can be used for the biological control of rice sheath blight disease, to the bacteriostasis rate of the pathogen thanatephorus cucumeris(frank) donk Thanatephoruscucumeris of rice sheath blight disease up to 81.9%, control efficiency is up to 70.0% or more.The present invention carries out separation screening using rice sheath blight disease pathogen as controlling object, to Antagonistic Endophytic strain present in healthy plant rice, lays the foundation for the biological control of rice sheath blight disease.The reasonable application of Bacillus strain ZY122 provided by the invention can reduce environmental pollution caused by chemical agent; be conducive to protecting ecology balance; very big raising and facilitation can be played with volume increase is increased income to the artificial cultivation of rice, there is certain research significance.

Description

One plant have inhibit rice sheath blight disease endophytic Bacillus bacterial strain ZY122 and its Using
Technical field
The present invention relates to technical field of plant disease biological control, and in particular, to one plant has inhibition rice sheath blight disease Endophytic Bacillus bacterial strain ZY122 and its application.
Background technique
Rice is the staple food grain of global about 50% population as one of big staple food grain in the world three.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 world's Monitoring of Paddy Rice Plant Area at present, China Rice Cropping face are 31,000,000 hectares, account for about generation The 20% of boundary's cultivated area；The rice total output in China ranks first in the world, and per unit area yield occupies the world the 5th.Rice is mainly distributed in China The producing region Yu Liu great is respectively as follows: South China Double Cropping rice and makees band；East China, Central China list double rice cropping band；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 increasing, how to ensure that the balance of eating rice production and consumption proposes severe challenge to the development of China's rice industry. In China under the influence of freshwater resources anxiety, labor shortage and the factors such as the production cost increases, Rice Cropping increases grain and increases income Contradiction becomes increasingly conspicuous, and along with global climate and ecological environment are worsening, causes the various pest and disease damages of rice to present increased Situation.
Rice sheath blight disease is one of the Major Diseases in current Rice Production, which is by thanatephorus cucumeris(frank) donkThanatephoruscucumerisIt is caused, thanatephorus cucumeris(frank) donkThanatephoruscucumerisBelong to Basidiomycotina fungi, Occurrence scope is wide, harm is serious, prevention and treatment is difficult.The disease prevents 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 prevention and treatment of rice sheath blight disease, also there is emerging biological Control Technology.The country is related to rice line There are about 400 remainders for the patented technology of blight, and wherein the relevant patent of chemical agent accounts for about 60%；Biotechnology accounts for about 30%；Chemistry The patented technology combined with both biological controls accounts for about 10%.
Bacillus has the characteristics that breeding is quick, metabolism is fast universally present in nature；The important spy of the category bacterium Property be can generate to unfavorable conditions have special resistance gemma.They exist in soil, in plant tissue, water and sky It is closely related with the conversion of nature substance, 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 released, synthesizes the substances such as a variety of organic acids, enzyme, physiological activity and other a variety of appearances The nutrient being easily utilized promotes plant growth.With the two packing spaces sexual involution and alkalization of soils of soil, bacillus can be It survives and grows in adverse circumstance, provide various enzymes for soil and increase the diversity of edaphon.Endophyte of plant is certain Stage or whole stages move in fungi or bacterium 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 mentioned to plant For needed nutrient matter and some hormones, have to the growth and development, the invasion of resistance disease pest and poor environment etc. of plant extensive Biological action can also enhance Genes For Plant Tolerance adverse circumstance, the ability of disease and insect resistance, have become at present potential micro- in biological control Biological pesticide, yield increasing fungus are used as potential biological and ecological methods to prevent plant disease, pests, and erosion carrier bacterium.
Endophyte exists in plant, and can colonize and conduct for a long time in plant, not vulnerable to external environmental condition It influencing, the bioactive substance of the slave endophyte of plant separation of studies have shown that 51% is the compound not found in the past, this To there is very great economic significance to the exploitation of the frontier in antibacterial substance source.Screening can have pathogen short of money The bacillus of anti-effect is in terms of the biological control of disease, promotion plant growth, improving with important Meaning.
Summary of the invention
Have it is an object of the invention to overcome the deficiencies of the prior art and provide one plant and inhibits to sprout in rice sheath blight disease Spore bacillus strain ZY122, endophytic Bacillus bacterial strain ZY122 provided by the invention can be used for inhibiting rice sheath blight disease, to water The pathogen thanatephorus cucumeris(frank) donk of the sheath and culm blight of riceThanatephoruscucumerisBacteriostasis rate up to 81.9%, control efficiency is reachable 70.0% or more.
It is anti-in the biology of rice sheath blight disease that another object of the present invention is to provide above-mentioned endophytic Bacillus bacterial strain ZY122 Application in controlling.
Another object of the present invention is to provide endophytic Bacillus bacterial strain ZY122 to prepare the life for inhibiting rice sheath blight disease Application in object preparation.
Another object of the present invention is to provide a kind of with the biological agent for inhibiting rice sheath blight disease.
To achieve the above object, the present invention adopts the following technical scheme:
One plant has the endophytic Bacillus bacterial strain ZY122 (Bacillus for inhibiting rice sheath blight disease VanilleaZY122), the bacterial strain is preserved in Guangdong Province's Culture Collection (GDMCC) on April 11st, 2017, Preservation address is Xianlie Middle Road, Guangzhou City 100, deposit number GDMCCNo.60161.
Heretofore described bacterial strain contains the 16SrDNA gene order as shown in SEQ1.
Bacillus of the inventionBacillus vanilleaZY122 colony morphology characteristic: the bacterial strain is on LB culture medium The speed of growth is very fast (37 DEG C, 20h), is in greyish white shape, is mucus type bacterial strain, and cell is sub-translucent, in (long 1.3~1.8um wide 0.4 ~0.7um) it is rod-shaped, there is protrusion among bacterium colony, gemma, peritrichous can be generated.
Bacillus of the inventionBacillus vanilleaThe physio-biochemical characteristics of ZY122:
Gram-positive bacteria, amphimicrobian, pH growth scope pH4.0~8.0(most preferably 5.0~7.0), resistance to NaCl can be (most preferably 1~4%) can be grown under the conditions of 0~8% (W/V), being suitable for growth temperature section is 20~45 DEG C (most preferably 28~7 DEG C), the G+C content of DNA is 46.4mol%.Produce ammonia test, acetyl formaldehyde Methanol determination is the positive；Urinate enzyme test, methyl red Measurement, catalase test, citrate utilization test are feminine gender.Gentiobiose, α-D-lactose, β-methyl-can efficiently be utilized D-glucoside, glycogen, D- xylose are then given birth to as carbon source using arabinose, melibiose, trehalose and turanose as carbon source It is long slow.Bacterial strain all has tolerance to 50 μ g/mL ampicillins, 100 μ g/mL streptomysins, 300 μ g/mL tetracyclines；And it is right Kalamycin, gentamicin, chloramphenicol, neomycin, erythromycin poor resistance, low concentration (5 μ g/mL) these types of antibiotic just It can inhibit its growth.
Bacillus provided by the inventionBacillus vanilleaZY122 is through 16SrDNA sequence alignment (EZBiocloud database) withBacillusvanilleaThe homology of XY18 (T) bacillus is 99.42%；The bacterial strain 16SrDNA sequence is as shown in sequence table.
Bacillus of the present inventionBacillus vanilleaThe determination of the molecular classification status of ZY122:
It is expanded using bacteria 16 S rRNA genes universal primer 25f and 1492r, PCR product is directly subjected to sequence survey It is fixed.The DNA sequence dna of acquisition input EZBiocloud is compared, primarily determine category of the bacterial strain of the invention in taxology, The position of kind.As a result, it has been found that bacillus of the invention belongs to bacillusBacillus, withBacillusvanilleaThe homology of XY18 (T) bacillus is 99.42%, but the sequence of the two is not quite identical, has 0.58% difference shows that the two is interior different bacterial strain of the same race.In conjunction with above-mentioned physio-biochemical characteristics, 16SrDNA sequence point Analysis, bacillus of the inventionBacillus vanilleaZY122 should belong to bacillusBacillus。
Bacillus of the inventionBacillus vanilleaZY122 can be used for the biological control of rice sheath blight disease.More Body, the Bacillus strain ZY122 can be applied to inhibit rice sheath blight disease pathogen thanatephorus cucumeris(frank) donkThanatephoruscucumeris.BacillusBacillus vanilleaZY122 dies 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 diseaseThanatephoruscucumerisControl efficiency is up to 70.0% or more.
Application of the above-mentioned endophytic Bacillus bacterial strain ZY122 in the biological agent that preparation inhibits rice sheath blight disease is also at this Within the protection scope of invention.
It is a kind of with the biological agent for inhibiting rice sheath blight disease, containing in described in claim 1 in the biological agent Raw Bacillus strain ZY122.
Compared with prior art, the invention has the following beneficial effects:
The present invention is right using the pathogen thanatephorus cucumeris(frank) donk of rice sheath blight disease as controlling object by using selective medium Antagonistic Endophytic present in the long-term unmanured wild rice plants of health is grown on to be separated, purified, obtain one plant it is short of money The bacillus of the water resistant sheath and culm blight of rice is laid a good foundation for the biological control of rice sheath blight disease.Endogenous spore provided by the invention The reasonable application of bacillus strain ZY122 can reduce chemical agent pollution on the environment, be conducive to protecting ecology balance, right The cultivation of rice and increase income volume increase can play very big raising and facilitation, have certain research significance.
Detailed description of the invention
Fig. 1 is the bacillus that embodiment 1 providesBacillus vanilleaZY122 dilution spread on LB culture medium Bacterium colony picture；
Fig. 2 is the pathogen thanatephorus cucumeris(frank) donk of rice sheath blight diseaseThanatephoruscucumerisControl group is cultivated in PDA Growth figure on base；
Fig. 3 is the bacillus that embodiment 1 providesBacillus vanilleaZY122 is to pathogen thanatephorus cucumeris(frank) donkThanatephoruscucumerisAntagonistic effect figure；
Fig. 4 is the bacillus that embodiment 1 providesBacillus vanilleaZY122 complete genome DNA and 16SrDNA PCR product 1% agarose gel electrophoresis figure.
Specific embodiment
Below with reference to chart, further description of the technical solution of the present invention, but this explanation does not do any form side The restriction in face, all modifying or equivalently replacing the technical solution of the present invention, without departing from the essence of technical solution of the present invention Mind and range, should all cover within the protection scope of the present invention.
The separation of 1 bacillus vanilleaZY122 of embodiment, screening and chemotaxonomy status Determine that the biocontrol bacterial strain bacillus vanilleaZY122 provided by the embodiment for inhibiting rice sheath blight disease is preserved in Guangdong Province's Culture Collection (GDMCC), preservation address Xianlie Middle Road, Guangzhou City 100, the deposit date is 2017 April 11, deposit number GDMCCNo.60161.
The bacillus of present embodimentBacillusZY122 derives from the wild rice field in Guangxi Wuzhou, using selective training It supports base and endogenetic bacteria separation acquisition is carried out to wild rice healthy plant, the specific method is as follows:
(1) separation of wild rice endophyte
The wild rice material of acquisition is cleaned with distilled water, suitable, stem, leaf texture is chosen, is cut into sterile scissors Fragment shape, is put into sterile petri dish, impregnates 3min with 75% ethyl alcohol, 3min is then impregnated in 2%NaClO solution, take out It uses aseptic water washing 3 times, each 5min afterwards, the sterile water of last time cleaning is coated with LB, and as control test, whether material disappears It is malicious thorough.Sterile water is added in sterile mortar and is ground into homogenate, after ten times of homogenate dilution, is drawn respectively with liquid-transfering gun On 100uL to sterile LB solid plate, 37 DEG C of constant incubator cultures are put into after smearing uniformly, observe the growing state of thallus, Lawns good with oese picking growing way and that form is different, carry out secondary culture repeatedly with plate streak, until the face of bacterium colony Color, shape, size, quality are as transparency.Finally by simple dyeing (dyeing of carbolic acid azaleine) and sediments microscope inspection (oil Mirror), its form is further looked at, equivalent width uniform with length, staining conditions are unified for bacterial strain purification standard.After purification Bacterial strain saves for -20 DEG C and -80 DEG C respectively in 15% glycerol.
(2) primary dcreening operation of endophyte antagonism rice banded sclerotial blight pathogen
Primary dcreening operation is carried out to wild rice endophyte antagonism banded sclerotial blight disease fungus using tablet face-off method.By rice sheath blight disease disease It is 10 that concentration, which is made, with sterile water in opportunistic pathogen8/mL-1Bacteria suspension, take 20uL plate to be coated in PDA culture medium, be then coated with PDA plate on mark signature line separate 4 regions, the heart is accessed with sterile toothpick be purified on a small quantity respectively in each area Wild rice endophyte is put in 28 DEG C of constant incubators and cultivates, the presence or absence of inhibition zone and size are observed and recorded after 5 days, and altogether repeatedly 3 It is secondary.It selects the maximum 5 plants of endophytes of inhibition zone width and carries out secondary screening.
(3) secondary screening of endophyte antagonism rice banded sclerotial blight pathogen
Secondary screening is carried out to endophyte antagonizing pathogenic fungi using tablet face-off method.By rice sheath blight disease pathogen sterile water It is 10 that concentration, which is made,8/mL-1Bacteria suspension, then 20uL is taken to be coated in PDA culture medium, the mark signature line on having been coated with plate 4 regions are separated, each regional center accesses preliminary screening with sterile toothpick respectively and comes out the endophyte for having antagonism, if nothing Bacterium water is control, is repeated 3 times.It is placed under 28 DEG C of constant temperatures and cultivates, fungistatic effect is observed after 5 days, measure inhibition zone size.Choosing One plant of best bacterium of effect out, saves backup.
Bacteriostasis rate (%)=(processing group antibacterial circle diameter-control group antibacterial circle diameter/processing group antibacterial circle diameter) × 100%
Culture medium prescription applied in the present embodiment is as follows:
LB culture medium: yeast extract 5g, bacteriological peptone 10g, Nacl5g, Agar20g, sterile water 1L, PH=7.0.
PDA culture medium: peeled potatoes 200g, glucose 20g, Agar20g, sterile water 1L, natural PH.
Fig. 1 is bacillus provided in this embodimentBacillus vanilleaZY122 dilution spread on LB culture medium Bacterium colony picture, left figure be the bacterium colony back side shine, right figure be bacterium colony front shine.Bacillus of the invention can be observed by Fig. 1Bacillus vanilleaZY122 colony morphology characteristic: bacterial strain speed of growth on LB culture medium is very fast (37 DEG C, 20h), It is mucus type bacterial strain in greyish white shape, cell is sub-translucent, and it is rod-shaped in (long 1.3~1.8um wide, 0.4~0.7um), have among bacterium colony Protrusion can generate 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 are thanatephorus cucumeris(frank) donk front photograph, and right figure is thanatephorus cucumeris(frank) donk back side photograph；Fig. 3 is bacillus VanilleaZY122 to the fungistatic effect figure of the pathogen thanatephorus cucumeris(frank) donk Thanatephoruscucumeris of rice sheath blight disease, Left figure is that antibacterial front is shone, and right figure is antibacterial back side photograph.BacillusBacillusPathogen melon of the ZY122 to rice sheath blight disease Die lead fungiThanatephoruscucumerisBacteriostasis rate up to 81.9%.
(4) bacillusBacillus vanilleaThe determination of the chemotaxonomy status of ZY122
Picking antagonistic BacillusBacillus vanilleaZY122 is inoculated in LB solid medium, is collected after 12h Thallus simultaneously extracts strain gene group DNA, with the size of 1% agarose electrophoresis validating DNA group segment.Fig. 4 is provided in this embodiment BacillusBacillus vanilleaThe agarose gel electrophoresis of the PCR product 1% of ZY122 complete genome DNA and 16SrDNA Figure, wherein left figure is 1% agarose gel electrophoresis figure of complete genome DNA, and right figure is the agar of the PCR product 1% of 16Sr DNA Sugared 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 greater than 10000bp； The electrophoretic band of PCR product in right figure is single clear, and fragment length is about 1500bp or so.
Using genomic DNA as template, using bacterial 16 S rDNA gene universal primer: 5 '-AGAGTTTGATC- of upstream primer 5 '-AAGGAGGTGATCCAGCC-3 of CTGGCTCAG-3 ' and downstream primer；
PCR reaction system: 10 × PCRbuffer2.5 μ L, dNTP2.0 μ L, each 0.5 μ L of upstream and downstream primer, DNA profiling 1.0 μ L, 0.2 μ L of Taq DNA polymerase mend distilled water to 25 μ L.PCR reaction condition: 94 DEG C of initial denaturation 5min；94 DEG C of denaturation 45s, 55 DEG C of annealing 45s, 72 DEG C of extension 1.5min, cyclic amplification 30 times: last 72 DEG C of extensions 10min.With 1% agarose electrophoresis Verify the size of PCR product DNA fragmentation, as shown in Figure 4, bacillusBacillus vanilleaZY122 DNA PCR product It is expanded by gel electrophoresis to single band, band is clear, and size is about 1500bp.Beijing is entrusted to hold up section's new industry PCR product Bioisystech Co., Ltd is sequenced, and the DNA sequence dna input GenBank of acquisition carries out Blast comparison, primarily determines the present invention BacillusBacillus vanilleaThe position of genus and species of the ZY122 bacterial strain in taxology.As a result, it has been found that the bud Spore bacillusBacillus vanilleaZY122, withBacillusvanilleaThe homology of XY18 (T) bacillus is 99.42%, it is of the invention in conjunction with above-mentioned morphological feature, physio-biochemical characteristics, 16SrDNA Phylogenetic Analysis and Literature Consult BacillusBacillus vanilleaZY122 should belong to bacillusBacillus。
Sequencing result is as follows:
GCTATAATGCAGTCGAGCGGACAGATGGGAGCTTGCTCCCTGATGTTAGCGGCGGACGGGTGAGTAACA CGTGGGTAACCTGCCTGTAAGACTGGGATAACTCCGGGAAACCGGGGCTAATACCGGATGGTTGTCTGAACCGCATG GTTCAGACATAAAAGGTGGCTTCGGCTACCACTTACAGATGGACCCGCGGCGCATTAGCTAGTTGGTGAGGTAACGG CTCACCAAGGCGACGATGCGTAGCCGACCTGAGAGGGTGATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCT ACGGGAGGCAGCAGTAGGGAATCTTCCGCAATGGACGAAAGTCTGACGGAGCAACGCCGCGTGAGTGATGAAGGTTT TCGGATCGTAAAGCTCTGTTGTTAGGGAAGAACAAGTGCCGTTCAAATAGGGCGGCACCTTGACGGTACCTAACCAG AAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGAATTATTGGGCGTAA AGGGCTCGCAGGCGGTTTCTTAAGTCTGATGTGAAAGCCCCCGGCTCAACCGGGGAGGGTCATTGGAAACTGGGGAA CTTGAGTGCAGAAGAGGAGAGTGGAATTCCACGTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCG AAGGCGACTCTCTGGTCTGTAACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGT CCACGCCGTAAACGATGAGTGCTAAGTGTTAGGGGGTTTCCGCCCCTTAGTGCTGCAGCTAACGCATTAAGCACTCC GCCTGGGGAGTACGGTCGCAAGACTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTT AATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCTCTGACAATCCTAGAGATAGGACGTCCCCTTCGGG GGCAGAGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAA CCCTTGATCTTAGTTGCCAGCATTCAGTTGGGCACTCTAAGGTGACTGCCGGTGACAAACCGGAGGAAGGTGGGGAT GACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGACAGAACAAAGGGCAGCGAAACCG CGAGGTTAAGCCAATCCCACAAATCTGTTCTCAGTTCGGATCGCAGTCTGCAACTCGACTGCGTGAAGCTGGAATCG CTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAGAGT TTGTAACACCCGAAGTCGGTGAGGTAACCTTTAGGAGCCAGCCGCCGAAG
Pathogen in the present embodiment is the thanatephorus cucumeris(frank) donk isolated from rice basal part of stemThanatephoruscucumeris。
Application of the 2 bacillus vanilleaZY122 of embodiment 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, Control efficiency experiment is as follows:
Three experimental groups are set and carry out potted plant experiments, respectively healthy control group, susceptible control group and prevention and treatment group, every group 20 Strain rice.Use thanatephorus cucumeris(frank) donkThanatephoruscucumerisBeing configured to concentration is 1 × 108The spore suspension of a/mL.It will After rice carries out surface sterilization, 20 μ L sterile waters, susceptible control group and each note of prevention and treatment group are injected to healthy control group with needle point method It is inoculated in flowerpot after entering 20 μ L thanatephorus cucumeris(frank) donk suspension.And the culture of 3mL antagonistic Bacillus is poured into prevention and treatment group plant root (concentration is 0.6 × 10 to liquid7~1 × 109cfu/L).Guarantee that the rice of each experimental group is grown at shady and cool ventilation, carries out daily It pours, observes and records rice pathogenesis situation after 15d.
Control efficiency=(susceptible group of plant number-control group disease plant number-prevention and treatment group disease plant number)/inoculation plant number × 100%.
1 bacillus of tableBacillus vanilleaZY122 inhibits thanatephorus cucumeris(frank) donkThanatephoruscucumeris Test
According to experimental result it is found that bacillusBacillus vanilleaZY122 is to thanatephorus cucumeris(frank) donkThanatephoruscucumerisControl efficiency up to 70.0% or more.
SEQUENCE LISTING
<110>Agricultural University Of South China
<120>one plants have the endophytic Bacillus bacterial strain ZY122 for inhibiting rice sheath blight disease and its application
<130> 1
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1428
<212> DNA
<213>16srDNA of bacillus vanilleaZY122
<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 (5)

1. one plant inhibition rice sheath blight disease endophytic Bacillus (Bacillus vanillea) bacterial strain ZY122, feature exists In the bacterial strain is preserved in Guangdong Province's Culture Collection on April 11st, 2017, and deposit number is GDMCC No: 60161。
2. application of the endophytic Bacillus bacterial strain ZY122 in the biological control of rice sheath blight disease described in claim 1.
3. applying according to claim 2, which is characterized in that the application is that Bacillus strain ZY122 is inhibiting rice Banded sclerotial blight pathogen thanatephorus cucumeris(frank) donkThanatephorus cucumerisIn application.
4. the answering in the biological agent that preparation inhibits rice sheath blight disease of endophytic Bacillus bacterial strain ZY122 described in claim 1 With.
5. a kind of biological agent for inhibiting rice sheath blight disease, which is characterized in that containing described in claim 1 in the biological agent Endophytic Bacillus bacterial strain ZY122.
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CN110628681B (en) * 2019-10-11 2021-04-30 沈阳农业大学 Bacillus altitudinis strain and application thereof in rice sheath blight
CN110878268B (en) * 2019-11-26 2021-07-20 湖南新九方科技有限公司 Bacillus megaterium for resisting rice sheath blight and application thereof

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