CN108641981A - One plant of Biocontrol Bacillus identification and application - Google Patents
One plant of Biocontrol Bacillus identification and application Download PDFInfo
- Publication number
- CN108641981A CN108641981A CN201810470743.3A CN201810470743A CN108641981A CN 108641981 A CN108641981 A CN 108641981A CN 201810470743 A CN201810470743 A CN 201810470743A CN 108641981 A CN108641981 A CN 108641981A
- Authority
- CN
- China
- Prior art keywords
- bacillus
- biocontrol
- yjc
- seq
- wheat
- 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
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/689—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for bacteria
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Analytical Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Agronomy & Crop Science (AREA)
- Pest Control & Pesticides (AREA)
- Plant Pathology (AREA)
- Dentistry (AREA)
- Environmental Sciences (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses one plant of Biocontrol Bacillus identification and applications, belong to field of agricultural microbial technology.The Biocontrol Bacillus of the present invention is Bei Laisi bacillus (Bacillus velezensis) YJC 9, which was preserved in Wuhan University's China typical culture collection center on 04 11st, 2018, and deposit number is CCTCC NO：M 2018196；The biocontrol microorganisms can prevent wheat sharp eyespot, wheat scab, sclerotinia sclerotiorum, grey mould fruit rot of strawberry, Fusarium solani, Kidney bean droop, tobacco brown spot pathogen, it is a kind of biocontrol bacteria with resistance of wide spectrum, has a good application prospect in agricultural disease prevention.
Description
Technical field
The invention belongs to field of agricultural microbial technology, relate generally to one plant of Bei Laisi bacillus and its Rapid identification side
Method and application in prevention wheat sharp eyespot etc..
Background technology
Wheat is China's staple food crop, and annual cultivated area is between hundred million mu of 3-4, and total output is more than 100,000,000 tons, position
It ranks the first in the world.In recent years, as wheat area constantly expands, commodity globalization, Cultivar replacing slow down, agricultural cultivating mode and
The factors such as high-yield culture technique popularization, wheat sharp eyespot is in China's wheat planting district occurrence degree getting worse, especially with wheat master
The producing region North China Plain, the Yangtze river basin and the ground such as river is expensive occur frequently and in aggravating trend year by year, when serious sick seedling up to 50% with
On, production loss 30-40%, occurring area has as many as tens million of mus, and very big influence is caused safely to the grain-production in China.
Wheat sharp eyespot is a kind of soil-borne disease, and the disease is mainly by soil-borne disease fungal pathogens Rhizoctonia cerealis (Rhizoctonia
Cerealis) cause with Rhizoctonia solani Kuhn (Rhizoctonia solani), in each stage of wheat entire breeding time
It causes damages, the symptoms such as withered rotten bud, sick seedling, the rotten stem of flower stalk, withered strain dead ears occurs in wheat after being infected.The evil of being critically ill is extensive,
In addition to endangering wheat, barley, rye, sorghum, rice, corn, cotton, tobacco, soybean, peanut, pakchoi, sesame can be also endangered
Etc. multiple kinds of crops.
Currently, chemical agent is still to prevent the most effective means of wheat sharp eyespot in production, mainly by dressing seed and
Systemic fungicide sprays to solve the problems, such as.But the long-term application of chemical agent, drawback and problem are increasingly prominent, and pathogen goes out
The risk that drug resistance is not only further exacerbated by chemical pesticide dosage is showed, and chemical agent causes environmental pollution and soil hardening
Acidification, and expection has also been well beyond to the harm of human health.Therefore, screen and study the environmentally friendly life of high activity
Anti- microbial inoculum prevents biology or abiotic stress, be solve chemical pesticide dosage for a long time remain high, Practice for Pesticide Residue in Agricultural Products
Effective way and China ensure grain quality safety, ecological environment security and reduce pollution of area source and protection bio-diversity
Effective way, be to realize the effective research direction such as " double to subtract ".
Invention content
In view of the deficienciess of the prior art, first of the present invention is designed to provide one plant to diseases such as Rhizoctonia cerealis
Opportunistic pathogen has the biocontrol microorganisms of strong antagonism.
In order to achieve the object of the present invention, inventor passes through many experiments and unremitting effort, obtains one plant of biological and ecological methods to prevent plant disease, pests, and erosion finally
Bacillus, the bacterial strain are isolated and purified from wheat healthy plant rhizosphere soil and are obtained, according to its colony morphology characteristic, 16srDNA,
GryA and rpoB Molecular Identifications and Phylogenetic analysis are quick and precisely identified as Bei Laisi bacillus (Bacillus
Velezensis), which has good inhibiting effect to diseases such as wheat sharp eyespots.
The isolated Bei Laisi bacillus of the present invention from it has been reported that Bei Laisi bacillus unlike, this hair
Bright is a kind of wide spectrum biocontrol microorganisms detached from healthy wheat rhizosphere soil, has to various agricultural pathogenic bacteria and inhibits to make well
With, including rhizoctonia cerealis, fusarium graminearum, Sclerotinia sclerotiorum, Botrytis cinerea germ, Fusarium Solani, Kidney bean
Wilt, tobacco brown spot pathogen.
Isolated Biocontrol Bacillus is named as Bei Laisi bacillus (Bacillus by the present inventor
Velezensis) YJC-9, the bacterial strain were preserved in Hongshan District Wuhan University of Wuhan City of Hubei China province on 04 11st, 2018
China typical culture collection center, deposit number are CCTCC NO：M 2018196.
It should be noted that Bei Laisi bacillus (Bacillus velezensis) YJC-9 that the present invention is isolated
With following morphological feature (Fig. 6)：
Morphological feature：Bei Laisi bacillus YJC-9 is milky, translucent, the smooth of the edge rule, thalline secretion
Can be in that cloud is spread, under the microscope, Gram's staining is the positive to oil, and rod-shaped, thalline is got together in catenation, and gemma is ellipse
Circle, middle life.
In addition, second object of the present invention is to provide the application of one plant of Biocontrol Bacillus, i.e., above-mentioned biological and ecological methods to prevent plant disease, pests, and erosion bud
Applications of spore bacillus Bei Laisi bacillus (Bacillus velezensis) YJC-9 in preventing agricultural disease.On and
State application of the Biocontrol Bacillus in the biological pesticide and/or biological organic fertilizer for preparing prevention agricultural disease.
Preferably, the agricultural disease include wheat sharp eyespot, wheat scab, sclerotinia sclerotiorum, grey mould fruit rot of strawberry,
Fusarium solani, Kidney bean droop and tobacco brown spot pathogen.
Finally, third object of the present invention is to provide the rapid identification method of one plant of Biocontrol Bacillus, this method
It is carried out by 16srDNA combinations gryA and rpoB gene pairs Bei Laisi bacillus (Bacillus velezensis) YJC-9
PCR amplification is identified；
Wherein 16srDNA primers are：27bf 5’-AGAGTTTGATCCTGGCTCAG-3’；
1492br 5’-GGTTACCTTGTTACGACTT-3’；
GyrA primers are：gyrA-42f*5’-CAGTCAGGAAATGCGTACGTCCTT-3’；
gyrA-1066r*5’-CAAGGTAATGCTCCAGGCATTGCT-3’；
RpoB primers are：rpoB-2292f 5’-GACGTGGGATGGCTACAACT-3’；
rpoB-3354r 5’-ATTGTCGCCTTTAACGATGG-3’；
PCR conditions are：94℃3min；94 DEG C of 45s, 55 DEG C of 30s, 72 DEG C of 2min, 35 cycles；72 DEG C extension 5min, 4 DEG C
Stop.
Compared with prior art, advantages of the present invention and marked improvement are：
1, Bei Laisi bacillus YJC-9 has different degrees of suppression to 7 kinds of disease fungus including Rhizoctonia cerealis
Effect processed is a kind of biocontrol bacteria with resistance of wide spectrum, is had a good application prospect in agricultural disease prevention.
2, Bei Laisi bacillus YJC-9 pot experiments are relatively anti-to inhibiting wheat sharp eyespot to have good control effect
Effect reaches 86.15%, and disease alleviates 65.67%, can be applied to the prevention of wheat sharp eyespot, is conducive to the grain in China --- and it is small
The safety in production of wheat.
Description of the drawings
Fig. 1 is Bei Laisi bacillus YJC-9 to rhizoctonia cerealis tablet fungistatic effect, and the left side is Bei Laisi gemma bars
For bacterium YJC-9 to rhizoctonia cerealis tablet fungistatic effect, the right is control.
Fig. 2 is the 16S rDNA gene order phylogenetic trees of Bei Laisi bacillus YJC-9.
Fig. 3 is the gryA gene order phylogenetic trees of Bei Laisi bacillus YJC-9.
Fig. 4 is the rpoB gene order systematic evolution trees of Bei Laisi bacillus YJC-9.
Fig. 5 is the inhibition to 6 kinds of disease fungus of Bei Laisi bacillus YJC-9, and wherein A, C, E, G, I, K distinguishes
For YJC-9 to Fusarium graminearum, Sclerotinia sclerotiorum, Botrytis cinerea,
Fusarium solani, Fusarium oxysporum, Alternaria alternata antagonistic effects；B, D, F, H, J, L according to
Secondary is above-mentioned control pathogen.
Fig. 6 is the morphological feature of Bei Laisi bacillus YJC-9, and left figure is that Bei Laisi bacillus YJC-9 is trained in NA
The colonial morphology grown on base is supported, right figure is the form of oily microscopic observation.
Specific implementation mode
The following is specific embodiments of the present invention, is described in detail to technical scheme of the present invention, but is not limited to
Protection scope of the present invention.
Embodiment 1：The separation of bacterial strain and antagonism detection
The health Shiyan City towns Liu Liping wife's parents' home river village school wheat rhizosphere soil 10g is collected, is put after being ground with sterilized mortar
Enter and shake suspension 10min in the triangular flask equipped with 90ml aseptic deionized waters, absorption 1ml suspensions, which are put into, from triangular flask is equipped with
Continue to shake suspension 5min in the sterile tube of 9ml aseptic deionized waters, suspension is then pressed into 10 times of further diluted concentrations of gradient
It is 10-4、10-5、10-6Three concentration.The suspension of these three concentration is put into baking oven and is arranged 80 DEG C, 15min selects can to produce
Sprout the bacterium of spore.
The liquid of three concentration is drawn 0.1ml and is put into respectively and has added the 1/10TSA tablets of ring acetimide (50 μ g/ml)
On smeared with spreading rod uniform, each concentration respectively applies three tablets.The tablet coated is put into 30 DEG C of cultures in constant incubator
3d.Rhizoctonia cerealis covers with 9mm culture dishes after PDA culture medium culture 5 days, with 5mm sterilization punchers in cultured cause of disease
Punched on bacterium culture medium, bacteria cake chosen into new PDA plate center, the bacterial clump for then selecting form color different away from
It is inoculated with from bacteria cake 3cm, opposite culture is carried out in PDA culture medium, the PDA plate to be only inoculated with sheath blight fungus is control, sieve
Choosing has the preferable bacterium of antagonistic activity (Fig. 1), is named as YJC-9.
Embodiment 2：16srDNA, gryA and rpoB Rapid identification of bacterial strain YJC-9
It is found in recent research, Bacillus genes similarity is all very high, these species include bacillus subtilis
(Bacillus subtilis, Cohn, 1872), bacillus amyloliquefaciens (Bacillus amyloliquefaciens,
Priest et al., 1987), atrophy bacillus (Bacillus atrophaeus, Nakamura, 1989), Mo Haiwei buds
Spore bacillus (Bacillusmojavensis, Roberts et al., 1994), Bacillus sonorensis (Palmisano
Et al., 2001), Te Jila bacillus (Bacillus tequilensis, Gatson et al., 2006) and methyl battalion
Support type bacillus (BacillusvallismortisRoberts et al., 1996) and Bei Laisi bacillus
(Bacillus velezensisEt al., 2005a) etc., the 16srDNA similarities of these bacillus are all
99% or more.Research has shown that particularly important come detection and identification bacterial species with Pulsed-field gel electrophoresis method, gryA is bud
The most fast gene of evolution degree in spore Bacillus phyletic evolution, rpoB genes are also answered in the classification of bacillus age identification extensively
With.Therefore, very necessary to kind is belonged to by 16srDNA combination gryA and rpoB gene pairs YJC-9 Rapid identifications.
Picking single bacterium colony after bacterial strain YJC-9 scribing line cultures is put into the triangular flask equipped with NB fluid nutrient mediums first, so
Triangular flask is put into shaking table 28 DEG C afterwards, 170rmin-1Culture is for 24 hours.Using YJC-9 bacterium solutions as template, 16srDNA is respectively adopted
Primer (27bf5 '-AGAGTTTGATCCTGGCTCAG-3 '；5 '-GGTTA CCTTGTTACGACTT-3 ' of 1492br), gyrA draws
Object (gyrA-42f*5 '-CAGTCAGGAAATGCGTACGTCCTT-3 ', gyrA-1066r*5 '-
CAAGGTAATGCTCCAGGCATTGCT-3 '), rpoB primers (rpoB-2292f 5 '-GACGTGGGATGGCTACAACT-3 ',
RpoB-3354r 5 '-ATTGTCGCCTTTAACGATGG-3 ') carry out PCR amplification.
PCR conditions are：94℃3min；94 DEG C of 45s, 55 DEG C of 30s, 72 DEG C of 2min, 35 cycles；72 DEG C extension 5min, 4 DEG C
Stop.PCR product send Wuhan Qing Ke Bioisystech Co., Ltd to be sequenced, and is used in combination MEGA5.0 to carry out Phylogenetic analysis, as a result
See Fig. 2, Fig. 3, Fig. 4, global alignment as a result, it has been found that bacterial strain YJC-9 and Bacillus velezensis (EU138632.1) and
Bacillus velezensis (EU138839.1) similitude highest, it may be determined that bacterial strain YJC-9 is
Bacillusvelezensis, and it is officially named Bei Laisi bacillus (Bacillus velezensis YJC-9), the bacterium
Strain was preserved on 04 11st, 2018 in Hongshan District Wuhan University of Wuhan City of Hubei China province China typical culture collection
The heart, deposit number are CCTCC NO：M 2018196.
Embodiment 3：YJC-9 detects other plant pathogen inhibition
It will identify the plant pathogenic fungi rhizoctonia cerealis (Rhizoctonia cerealis), small of preservation in this laboratory
Wheat gibberellic hypha (Fusarium graminearum), Sclerotinia sclerotiorum (Sclerotinia sclerotiorum), strawberry
Ash arrhizus bacteria (Botrytis cinerea), Fusarium Solani (Fusarium solani), Kidney bean wilt
After (Fusarium oxysporum), tobacco brown spot pathogen (Alternaria alternata) reactivate, beaten with 5mm is sterile
Bacteria cake, is inoculated in PDA culture medium center, YJC-9 bacterial strains is inoculated at bacteria cake 3cm, to be only inoculated with disease by hole device punching respectively
The PDA culture medium of opportunistic pathogen is control, and antibacterial bandwidth of the YJC-9 Antagonistic Fungis to these types of pathogen is recorded after 7d, calculates growth suppression
Rate processed, the results are shown in Table 1 and Fig. 5.
Inhibiting effect of the table 1YJC-9 to disease fungus
From table 1 it follows that B.velezensis YJC-9 have a variety of pathogens different degrees of inhibition,
10mm is wherein reached to rhizoctonia cerealis, Sclerotinia sclerotiorum, Botrytis cinerea germ and the antibacterial bandwidth of Kidney bean wilt
More than, rhizoctonia cerealis, fusarium graminearum, Sclerotinia sclerotiorum, Botrytis cinerea germ and Kidney bean wilt are grown
Inhibiting rate reaches 60% or more, is a kind of biocontrol bacteria with broad-spectrum sterilization effect.
Embodiment 4：YJC-9 inducing wheats generate resistance to prevent wheat sharp eyespot
Picking YJC-9 bacterial strain single bacterium colonies are put into NB culture mediums, 28 DEG C of shake culture 60h, with centrifuge 5000rmin-1, 10min is centrifuged, collects thalline, then use MgSO4Buffer solution dilutes bacterial concentration to 1 × 107Cfu/ml chooses healthy wheat
Seed first uses 1.2%NaOCl solution disinfection 5min, then with after 70% ethanol disinfection 5min, then is cleaned three times with aqua sterilisa, most
Wheat seed is put into the culture dish of aseptic filter paper afterwards, appropriate amounts of sterilized water is added, 30 DEG C of light culture 3d carry out vernalization.It will germination
Consistent seedling replanting enters in the native basin of (121 DEG C, the 20min) matrix that sterilizes equipped with 150g, is then added in every basin soil
15mlYJC-9 bacteria suspensions after growing 7d, after the rhizoctonia cerealis in PDA culture medium is punched with 3mm aseptic card punch, are chosen
It takes bacteria cake to be tightly attached to wheat basal part of stem to be inoculated with and fixed.To be only inoculated with rhizoctonia cerealis, it is not added with the small of bacterium bacteria suspension
Wheat is control treatment.Per basin 10 plants of seedling, processing and control respectively there are three repeating, while need to spray daily sterile water to wheat into
Row moisturizing, having small water droplet freely to be fallen along stalk to spray blade is advisable.After 10d, processing group and control group morbidity are recorded in observation
Situation (table 2).Wheat sharp eyespot severity and disease index are calculated with reference to the People's Republic of China's agricultural industry criteria：Wheat line
Blight observes and predicts investigation specifications NY/T 614-2002.
Table 2YJC-9 is to wheat sharp eyespot potting resistant proof
From Table 2, it can be seen that YJC-9 bacterial strains are playing good control effect, compared with the control, phase to banded sclerotial blight
86.15% is reached to preventive effect, disease alleviates 65.67%.
Sequence table
<110>Academy of Agricultural Sciences of Shiyan City
<120>One plant of Biocontrol Bacillus and its application
<160> 6
<170> SIPOSequenceListing 1.0
<210> 1
<211> 20
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 1
agagtttgat cctggctcag 20
<210> 2
<211> 19
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 2
ggttaccttg ttacgactt 19
<210> 3
<211> 24
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 3
cagtcaggaa atgcgtacgt cctt 24
<210> 4
<211> 24
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 4
caaggtaatg ctccaggcat tgct 24
<210> 5
<211> 20
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 5
gacgtgggat ggctacaact 20
<210> 6
<211> 20
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 6
attgtcgcct ttaacgatgg 20
Claims (6)
1. one plant of Biocontrol Bacillus, it is characterised in that：The Biocontrol Bacillus is Bei Laisi bacillus (Bacillus
Velezensis) YJC-9, preserving number are CCTCC NO：M 2018196.
2. application of the Biocontrol Bacillus described in claim 1 in preventing agricultural disease.
3. application of the Biocontrol Bacillus as claimed in claim 2 in preventing agricultural disease, it is characterised in that：The agriculture
Industry disease include wheat sharp eyespot, wheat scab, sclerotinia sclerotiorum, grey mould fruit rot of strawberry, Fusarium solani, Kidney bean droop and
Alternaria alternate.
4. Biocontrol Bacillus described in claim 1 is in the biological pesticide and/or biological organic fertilizer for preparing prevention agricultural disease
In application.
5. Biocontrol Bacillus as claimed in claim 4 is in the biological pesticide and/or biological organic for preparing prevention agricultural disease
Application in fertilizer, it is characterised in that：The agricultural disease includes wheat sharp eyespot, wheat scab, sclerotinia sclerotiorum, strawberry
Gray mold, Fusarium solani, Kidney bean droop and Alternaria alternate.
6. a kind of identification method of Biocontrol Bacillus as described in claim 1, it is characterised in that：This method passes through
16srDNA combinations gryA and rpoB gene pairs Bei Laisi bacillus (Bacillus velezensis) YJC-9 carries out PCR expansions
Increase identification；Wherein, the primer pair of 16srDNA such as SEQ ID NO：1 and SEQ ID NO：Shown in 2, the primer pair such as SEQ of gyrA
ID NO：3 and SEQ ID NO：Shown in 4, the primer pair such as SEQ ID NO of rpoB：5 and SEQ ID NO：Shown in 6, PCR conditions
For：94℃3min；94 DEG C of 45s, 55 DEG C of 30s, 72 DEG C of 2min, 35 cycles；72 DEG C of extension 5min, 4 DEG C of stoppings.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810470743.3A CN108641981B (en) | 2018-05-17 | 2018-05-17 | Biocontrol bacillus identification and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810470743.3A CN108641981B (en) | 2018-05-17 | 2018-05-17 | Biocontrol bacillus identification and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108641981A true CN108641981A (en) | 2018-10-12 |
| CN108641981B CN108641981B (en) | 2020-10-02 |
Family
ID=63756211
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810470743.3A Expired - Fee Related CN108641981B (en) | 2018-05-17 | 2018-05-17 | Biocontrol bacillus identification and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108641981B (en) |
Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109554317A (en) * | 2018-12-29 | 2019-04-02 | 陕西省微生物研究所 | Bei Laisi bacillus and its application in wheat sharp eyespot disease prevention growth-promoting |
| CN110283742A (en) * | 2019-06-17 | 2019-09-27 | 北京农业生物技术研究中心 | The Bei Laisi bacillus of one plant of broad-spectrum disease resistance and its application |
| CN111100809A (en) * | 2019-12-17 | 2020-05-05 | 新疆农业科学院核技术生物技术研究所（新疆维吾尔自治区生物技术研究中心） | Bacillus mojavensis for preventing and treating root rot of cotton in seedling stage and application thereof |
| CN111534460A (en) * | 2020-04-28 | 2020-08-14 | 湖北大学 | Preparation method and application of bacillus beiLeisi capable of efficiently antagonizing fusarium graminearum |
| CN111747789A (en) * | 2020-07-08 | 2020-10-09 | 云南省烟草公司曲靖市公司 | Biological foliar fertilizer with functions of preventing and treating powdery mildew and alternaria alternate, and preparation method and application thereof |
| CN111748498A (en) * | 2020-07-08 | 2020-10-09 | 云南省烟草公司曲靖市公司 | Bacillus compound microbial inoculum with functions of preventing and treating powdery mildew and brown spot as well as preparation method and application thereof |
| CN111887261A (en) * | 2020-09-01 | 2020-11-06 | 天津开发区坤禾生物技术有限公司 | Application of Bacillus belgii in prevention and treatment of sunflower sclerotium disease |
| CN112322515A (en) * | 2019-07-16 | 2021-02-05 | 浙江泛亚生物医药股份有限公司 | Novel strain of Bacillus belgii and application thereof |
| CN112980721A (en) * | 2021-01-29 | 2021-06-18 | 青岛农业大学 | Bacillus belgii and biocontrol preparation and application thereof |
| CN113151051A (en) * | 2021-03-15 | 2021-07-23 | 哈尔滨师范大学 | Bacillus belgii and application thereof |
| CN113980836A (en) * | 2021-09-23 | 2022-01-28 | 中化化肥有限公司临沂农业研发中心 | Bacillus belgii and application thereof in prevention and treatment of strawberry root rot |
| CN114107280A (en) * | 2021-11-22 | 2022-03-01 | 江苏科技大学 | Method for improving physiological and biochemical activities of strains through electrical stimulation and application of method |
| CN114196565A (en) * | 2021-10-14 | 2022-03-18 | 河南农业大学 | Bacillus atrophaeus XZRKZ-23 and application thereof |
| CN114369556A (en) * | 2022-02-10 | 2022-04-19 | 成都市农林科学院 | Bacillus, biocontrol microbial inoculum prepared from bacillus and application of biocontrol microbial inoculum |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008113873A1 (en) * | 2007-03-19 | 2008-09-25 | Probelte, Sa | A pure culture of strain ah2 of the bacillus velezensis species and a product for the biological control of phytopathogenic fungi |
| CN102703342A (en) * | 2012-04-25 | 2012-10-03 | 四川农业大学 | Bacillus velezensis ZJ20 strain and liquid preparations thereof |
| CN107446847A (en) * | 2017-08-14 | 2017-12-08 | 云南农业大学 | One plant of Bei Laisi bacillus GT11 and its application |
| CN107964514A (en) * | 2017-09-08 | 2018-04-27 | 华夏春秋科技发展（北京）有限责任公司 | A kind of Bei Laisi bacillus and its application on plant |
| CN108004185A (en) * | 2018-01-09 | 2018-05-08 | 中国农业科学院植物保护研究所 | One plant of tool diseases prevention, growth-promoting, drought resisting function plant endogenesis Bei Laisi bacillus and its application |
-
2018
- 2018-05-17 CN CN201810470743.3A patent/CN108641981B/en not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008113873A1 (en) * | 2007-03-19 | 2008-09-25 | Probelte, Sa | A pure culture of strain ah2 of the bacillus velezensis species and a product for the biological control of phytopathogenic fungi |
| CN102703342A (en) * | 2012-04-25 | 2012-10-03 | 四川农业大学 | Bacillus velezensis ZJ20 strain and liquid preparations thereof |
| CN107446847A (en) * | 2017-08-14 | 2017-12-08 | 云南农业大学 | One plant of Bei Laisi bacillus GT11 and its application |
| CN107964514A (en) * | 2017-09-08 | 2018-04-27 | 华夏春秋科技发展（北京）有限责任公司 | A kind of Bei Laisi bacillus and its application on plant |
| CN108004185A (en) * | 2018-01-09 | 2018-05-08 | 中国农业科学院植物保护研究所 | One plant of tool diseases prevention, growth-promoting, drought resisting function plant endogenesis Bei Laisi bacillus and its application |
Non-Patent Citations (1)
| Title |
|---|
| 蔡高磊等: "贝莱斯芽孢杆菌(Bacillus velezensis)研究进展 ", 《北方园艺》 * |
Cited By (19)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109554317A (en) * | 2018-12-29 | 2019-04-02 | 陕西省微生物研究所 | Bei Laisi bacillus and its application in wheat sharp eyespot disease prevention growth-promoting |
| CN110283742A (en) * | 2019-06-17 | 2019-09-27 | 北京农业生物技术研究中心 | The Bei Laisi bacillus of one plant of broad-spectrum disease resistance and its application |
| CN112322515A (en) * | 2019-07-16 | 2021-02-05 | 浙江泛亚生物医药股份有限公司 | Novel strain of Bacillus belgii and application thereof |
| CN111100809B (en) * | 2019-12-17 | 2023-06-06 | 新疆农业科学院核技术生物技术研究所（新疆维吾尔自治区生物技术研究中心） | Bacillus mojavensis for preventing and treating root rot of cotton at seedling stage and application thereof |
| CN111100809A (en) * | 2019-12-17 | 2020-05-05 | 新疆农业科学院核技术生物技术研究所（新疆维吾尔自治区生物技术研究中心） | Bacillus mojavensis for preventing and treating root rot of cotton in seedling stage and application thereof |
| CN111534460A (en) * | 2020-04-28 | 2020-08-14 | 湖北大学 | Preparation method and application of bacillus beiLeisi capable of efficiently antagonizing fusarium graminearum |
| CN111747789A (en) * | 2020-07-08 | 2020-10-09 | 云南省烟草公司曲靖市公司 | Biological foliar fertilizer with functions of preventing and treating powdery mildew and alternaria alternate, and preparation method and application thereof |
| CN111748498A (en) * | 2020-07-08 | 2020-10-09 | 云南省烟草公司曲靖市公司 | Bacillus compound microbial inoculum with functions of preventing and treating powdery mildew and brown spot as well as preparation method and application thereof |
| CN111887261A (en) * | 2020-09-01 | 2020-11-06 | 天津开发区坤禾生物技术有限公司 | Application of Bacillus belgii in prevention and treatment of sunflower sclerotium disease |
| CN112980721A (en) * | 2021-01-29 | 2021-06-18 | 青岛农业大学 | Bacillus belgii and biocontrol preparation and application thereof |
| CN112980721B (en) * | 2021-01-29 | 2022-06-10 | 青岛农业大学 | Bacillus belgii and biocontrol preparation and application thereof |
| CN113151051A (en) * | 2021-03-15 | 2021-07-23 | 哈尔滨师范大学 | Bacillus belgii and application thereof |
| CN113980836A (en) * | 2021-09-23 | 2022-01-28 | 中化化肥有限公司临沂农业研发中心 | Bacillus belgii and application thereof in prevention and treatment of strawberry root rot |
| CN113980836B (en) * | 2021-09-23 | 2022-09-09 | 中化化肥有限公司临沂农业研发中心 | Bacillus belgii and application thereof in prevention and treatment of strawberry root rot |
| CN114196565A (en) * | 2021-10-14 | 2022-03-18 | 河南农业大学 | Bacillus atrophaeus XZRKZ-23 and application thereof |
| CN114196565B (en) * | 2021-10-14 | 2024-03-01 | 河南农业大学 | Bacillus atrophaeus XZRKZ-23 and application thereof |
| CN114107280A (en) * | 2021-11-22 | 2022-03-01 | 江苏科技大学 | Method for improving physiological and biochemical activities of strains through electrical stimulation and application of method |
| CN114369556A (en) * | 2022-02-10 | 2022-04-19 | 成都市农林科学院 | Bacillus, biocontrol microbial inoculum prepared from bacillus and application of biocontrol microbial inoculum |
| CN114369556B (en) * | 2022-02-10 | 2023-07-04 | 成都市农林科学院 | Bacillus, biocontrol microbial agent prepared from bacillus and application of biocontrol microbial agent |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108641981B (en) | 2020-10-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108641981A (en) | One plant of Biocontrol Bacillus identification and application | |
| CN104498386B (en) | The preparation method and application of raw Bacillus amyloliquefaciens strain SZ23 and zymotic fluid in wild jujube | |
| CN105219681B (en) | A kind of bacillus amyloliquefaciens Bacillus amyloliquefaciens D2WM and preparation method and application | |
| CN109022315A (en) | The biological and ecological methods to prevent plant disease, pests, and erosion bacillus of one plant of broad-spectrum disease resistance and its application | |
| CN105543132B (en) | A kind of Methylotrophic bacillus YB-F7 and its application in controlling plant diseases | |
| CN104762223A (en) | Bacillus amyloliquefaciense BA-KA3 and application thereof | |
| CN103756931A (en) | Paenibacillus kribbensis and its application | |
| CN102433282A (en) | Bacillus subtilis NB12, as well as culture method and application thereof | |
| CN105441366B (en) | Methylotrophic bacillus ZBL-1 is applied in preventing cotton verticillium wilt | |
| CN106479934B (en) | A kind of ash arrhizus bacteria antagonistic strain and its screening technique and application | |
| CN109022320A (en) | Siam bacillus and its application in root rot and southern blight prevention and treatment | |
| CN114410481B (en) | Trichoderma asperellum and application thereof | |
| CN106635870A (en) | Bacillus and application thereof | |
| CN107099474A (en) | A kind of Pseudomonas chlororaphis and its application with broad spectrum antibiotic activity | |
| CN105018385A (en) | Bacillus amyloliquefaciens and application thereof | |
| CN105018386A (en) | Bacillus methylotrophicus and application thereof | |
| CN104651260A (en) | Brevibacillus brevis BBC-3 and application thereof as well as preparation method of microbial inoculum of brevibacillus brevis | |
| CN105018391B (en) | A kind of dead paddy bacillus NBIF 001 and fermentation process and application | |
| CN104974965A (en) | Bacillus subtilis JN005 and application thereof in prevention and control of rice blast | |
| CN106591157A (en) | Aspergillus tubingensis with disease prevention and growth promoting functions as well as preparation and application of aspergillus tubingensis metabolites | |
| CN106282049B (en) | Te Jila bacillus JN369 and its application | |
| CN103695360B (en) | The separation method of a kind of ocean bacillus amyloliquefaciens 1A and purposes | |
| CN111424004B (en) | Pseudomonas linusii and application thereof | |
| CN104789509B (en) | Raw bacillus pumilus and its application in one plant of bark of eucommia | |
| CN106701619A (en) | High-density fermentation method for bacillus amyloliquefaciens and preparation method of microbial agent of bacillus amyloliquefaciens |
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 | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20201002 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |