CN110257286A - One plant of bacillus amyloliquefaciens that can inhibit disease fungus - Google Patents
One plant of bacillus amyloliquefaciens that can inhibit disease fungus Download PDFInfo
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- CN110257286A CN110257286A CN201910531471.8A CN201910531471A CN110257286A CN 110257286 A CN110257286 A CN 110257286A CN 201910531471 A CN201910531471 A CN 201910531471A CN 110257286 A CN110257286 A CN 110257286A
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- 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
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- 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
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- C12N1/205—Bacterial isolates
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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Abstract
The invention discloses the bacillus amyloliquefaciens that one plant can inhibit disease fungus, its classification naming is bacillus amyloliquefaciens MT310 (Bacillus amyloliquefaciens MT310), it is preserved in China typical culture collection center, culture presevation number is CCTCC NO:M2019046, and the deposit date is on January 14th, 2019；The disease fungus is tobacco black shank bacterium, tobacco brown spot pathogen, Colletotrichum capsici and sorghum A. mali.
Description
Technical field
The present invention relates to the bacillus amyloliquefaciens that one plant can inhibit disease fungus.
Background technique
Tobacco black shank (Phytophthora nicotianae) is most destructive soil-borne disease in Guizhou Tobacco production
One of evil, nearly ten million yuan of the economic loss as caused by the disease, seriously affects the income of tobacco grower every year.At present in production
Chemical control is mostly used greatly, but primary medicament service life is longer, pathogen drug resistance increases year by year, and pesticide residue and environment are dirty
The negative effects such as dye also gradually increase.Therefore, carrying out tobacco diseases prevention and treatment using biological control is the hot fields studied at present.
Bacillus (Bacillus spp.) is most common microbe species in current biological control research, gemma bar
Bacterium has the prospect of industrialization, and mechanism of action multiplicity, production cost is low, and easy storage and transportation is not likely to produce drug resistance, to safety of human and livestock etc.
Advantage is the important directions of biological prevention and control agent exploitation.Native Bacillus separation strains excellent there are many character domestic and international at present
As living microorganism preparation viable commercial product application, development potentiality is huge.China earliest by China Agricultural University develop by
Yield increasing fungus made of the bacillus such as bacillus cereus (Bacillus cereus) and bacillus subtilis (B.subtilis)
Series of products have good volume increase, protection effect on rice, wheat, vegetables and more industrial crops.Gustafson company
The BioYield of research and development is by bacillus amyloliquefaciens (Bacillus amyloliquefaciens) FZB42 and bacillus subtilis
Fungicide made of (Bacillus subtilis) GB122 is mixed, has been applied to the prevention and treatment of plurality of plant diseases.
Based on above-mentioned basis, to reduce influence of the tobacco black shank to tobacco industry, a energy need to be researched and developed and be suitble to tobacco disease
Harmful biological bacteria preparation.
Summary of the invention
The technical problem to be solved in the present invention is to provide the bacillus amyloliquefaciens that one plant can inhibit disease fungus, can with gram
Take the deficiencies in the prior art.
Technical solution of the present invention:
One plant of bacillus amyloliquefaciens that can inhibit disease fungus, classification naming are bacillus amyloliquefaciens MT310
(Bacillus amyloliquefaciens MT310) is preserved in China typical culture collection center, culture presevation number
For CCTCC NO:M 2019046, the deposit date is on January 14th, 2019, address was Wuhan, China Wuhan University.
Can inhibit the bacillus amyloliquefaciens of disease fungus, the disease fungus be tobacco black shank bacterium, tobacco brown spot pathogen,
Colletotrichum capsici and sorghum A. mali.
Detailed description of the invention
Fig. 1 bacillus screens the antagonistic effect of tobacco black shank bacterium.
Fig. 2 bacterial strain MT310 biomembrane, protease, cellulase, thermophilic iron element and phosphate detection.
Fig. 3 bacterial strain MT310 16S rDNA phylogenetic trees.
The identification of Fig. 4 bacterial strain MT310 antimicrobial spectrum.
Its its classification naming of the microorganism of the application preservation is bacillus amyloliquefaciens MT310 (Bacillus
Amyloliquefaciens MT310), it is preserved in China typical culture collection center, culture presevation number is CCTCC NO:
M 2019046, the deposit date is on January 14th, 2019, address was Wuhan, China Wuhan University.
Specific embodiment
Embodiment 1, the acquisition of bacillus amyloliquefaciens MT310 and compliance test result
1 materials and methods
1.1 material
1.1.1 soil sample acquires
2017-2018 chooses the serious plot of tobacco black shank morbidity from Zunyi City, Guizhou Province Meitan County, acquires health-care cigarette
Same plot soil sample is mixed and calculates 1 part of soil sample by strain rhizosphere soil, 5 point samplings.Soil sample, soil are collected using sterile valve bag
Sampling instrument is carried out disinfection with 75% alcohol, and edaphon between different fields is avoided to mix.
1.1.2 culture medium
LB culture medium is used for the separation and preservation of bacillus；PDA culture medium is tested for plate opposite culture；Protease
Detection culture medium, cellulase detection culture medium, thermophilic iron element detection culture medium, phosphate detection culture medium are for detecting gemma
Whether bacillus generates protease, cellulase, thermophilic iron element and phosphate etc..
1.1.3 strains tested
Tobacco black shank bacterium (P.nicotianae) is provided by Plant Pathology teaching and research room, agricultural college, Guizhou University.
The separation of 1.2 rhizosphere bacillus and preservation
Using dilution-plate method, by soil dilution liquid, 30 min of water-bath is non-to kill the overwhelming majority in 85 DEG C of water-bath
Spore bacteria.The different single colonie of picking purifies bacterial strain and is stored in -80 DEG C of refrigerators.
The screening of 1.3 prevention and treatment tobacco black shank bacillus
The prevention and treatment apparent biocontrol bacterial strain of tobacco black shank effect is screened by tablet face-off method, is observed by primary dcreening operation and secondary screening
Inhibition zone whether is generated, and measures its bacteriostatic diameter using crossing method.Again by specific plate detected whether biomembrane,
The generation of protease and cellulase, wherein the best bacterial strain of 3 plants of effects is used for subsequent field experiment for selection.
Bacteriostasis rate (%)=[(control group colony diameter-processing group colony diameter)/processing group colony diameter] × 100.
The identification of 1.4 biocontrol bacterial strains is carried out by the bacterial strain of morphology, physiological and biochemical property and the screening of 16S rDNA gene pairs
Identification.
1.4.1 colony morphological observation
In inoculating strain to the test tube containing 5mL LB culture solution, 37 DEG C are shaken training overnight, and thalline were collected by centrifugation, thallus nothing
Bacterium water cleans 3 times, is finally resuspended with sterile water and is diluted to concentration 1 × 10810 μ L bacterium solutions are added dropwise in CFU/mL on LB plate,
37 DEG C be incubated overnight after observe colonial morphology.
1.4.2 physiological and biochemical property
According to " primary Jie Shi systematic bacteriology handbook ", the physio-biochemical characteristics of bacterial strain are measured.
1.4.3 molecular biology identification
After 37 DEG C of water-bath 1h processing thallus of 50mg/mL lysozyme, Biomiga bacterial genomes DNA extraction kit is used
It extracts.Using genomic DNA as template, PCR amplification is carried out using 16S rDNA universal primer 27F/1492R, PCR product is verified just
The raw work in sea is served after really carries out PCR product sequencing.Sequencing result carries out BLAST in ncbi database and compares analysis, and utilizes
CLUSTAL X software carries out the Neighbor-Joining algorithm structure in Multiple Sequence Alignment and phylogenetic analysis software MEGA6
Build phylogenetic tree.
1.5 field trials (only two plants of bacterium have been cooked crop field GUMT319 and GUMT323)
Experimental cultivar: cloud and mist 87.Test site: the Meitan County town Chao Le.Experimental field: fertility is uniform, plantation flue-cured tobacco in 2017
And balck shank morbidity plot.
Experimental design: the bacillus of screening is 1 processing, number MT310, with 30% first frost hymexazol water of medicament
Agent (middle peasant Li Hua (Tianjin) agricultural chemicals Co., Ltd) is positive control, and not carrying out any chemicals treatment is blank pair
According to, totally 5 processing, 3 repetitions, 30 plants of cigarettes of every cell, all random district's groups arrangements of processing.Chemicals treatment concentration and dosage reference
Medicament explanation, pouring root is applied when tobacco seedlings are transplanted, and is applied 2 times；Bacillus concentration is 108CFU/mL, 50mL bacterium solution pouring root are applied
With, co-administer 3 times, from tobacco seedlings transplant when pouring root 1 time, after every 7-10d pouring root 1 time.Transplanting on April 23rd, 2018, field plant
Training management method is consistent with locality, does not apply other fungicide.
1.6 field investigation
The field investigation time is chosen and carries out 3 times altogether after the onset of blank control starts to harvesting is preceding, and all processing cells are investigated
Cigarette strain balck shank disease incidence, and it is classified investigation disease severity.Tobacco black shank disease scale standard is according to GB/T 23222-
2008 carry out, and calculate the disease incidence and disease index of disease, data using DPS data processing system Duncan duncan's new multiple range method into
Row variance analysis.
2.2 complete technical solutions provided by the invention
The screening of 1 antagonism tobacco black shank bacterium bacillus
With tobacco black shank bacterium (P.nicotianae) be for trying pathogen, using tablet face-off method, through primary dcreening operation and secondary screening,
Obtain the preferable bacillus of antagonistic effect, number is MT310 (see Fig. 1).CK in Fig. 1: tobacco black shank bacterium
(Phytophthora nicotianae)；MT310 indicates the Bacillus strain with antagonistic effect screened.
2 bacterial strain biomembranes, protease and cellulase measurement
MT310 can produce more complicated biofilm structure, can produce protease and cellulase, it may be possible to which it inhibits true
One of mechanism of bacterium.It can produce phosphate and thermophilic iron element, promote plant growth related to bacillus.Bacillus is in plant
Root colonization simultaneously generates complicated biofilm structure, can effectively occupy root table Advantageous sites, limit the intrusion of pathogen mycelia,
And can induce plant and resistance is generated to pathogen, achieve the effect that disease-preventing and yield-increasing.
The identification of 3 biocontrol bacterial strains
MT310 can form more complicated colony structure (Fig. 2) on LB culture medium.The physio-biochemical characteristics of bacterial strain are shown in Table
1, MT310 is gram-positive bacterium, can decompose glucose, rhamnose and mannitol, generates catalase and nitric acid also
Protoenzyme, methyl red stained positive, the features such as liquefiable gelatin.By 16S rDNA gene sequencing, bacterial strain is constructed
The 16S rDNA phylogenetic trees of MT310, the results show that bacterial strain MT310 and bacillus amyloliquefaciens
(B.amyloliquefaciens) gather a branch (Fig. 3).Pass through morphology, physio-biochemical characteristics and 16S rDNA gene
Identify that GUMT310 is bacillus amyloliquefaciens (B.amyloliquefaciens).
1 bacterial strain MT310 physiological and biochemical property of table
In table 1+: positive reaction；: 4 field plot trial of negative reaction
Field diseases investigation is carried out when blank control cell tobacco black shank takes place.On the June 26th, 2018 of investigation the
It 1 time, investigates 1 time every 7-10d, investigates 3 times altogether.Table 2 is it is found that MT310 has certain preventive effect to tobacco black shank, but preventive effect is not
And first frost hymexazol.First frost hymexazol is for preventing and treating the primary medicament of tobacco black shank in current production, in this test
It is good to tobacco black shank control efficiency.
Field plot preventive effect of 22 bacillus of table to tobacco black shank
The identification of 5 biocontrol bacterial strain antimicrobial spectrums
From fig. 4, it can be seen that MT310 has wide spectrum antagonistic substance, to tobacco black shank bacterium, tobacco brown spot pathogen, capsicum anthrax
A variety of disease fungus fungistatic effects such as germ and sorghum A. mali are preferable.Show that this plant of bacterium has potential life to the above disease
Anti- effect.
Claims (2)
1. one plant of bacillus amyloliquefaciens that can inhibit disease fungus, classification naming is bacillus amyloliquefaciens MT310
(Bacillus amyloliquefaciens MT310) is preserved in China typical culture collection center, culture presevation number
For CCTCC NO:M 2019046, the deposit date is on January 14th, 2019, address was Wuhan, China Wuhan University.
2. the bacillus amyloliquefaciens according to claim 1 that disease fungus can be inhibited, it is characterised in that: the cause of disease
Fungi is tobacco black shank bacterium, tobacco brown spot pathogen, Colletotrichum capsici and sorghum A. mali.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110747208A (en) * | 2019-11-28 | 2020-02-04 | 海南大学 | Cassava nitrate reductase gene and construction and disease-resistant application of overexpression vector thereof |
| CN114250171A (en) * | 2021-12-08 | 2022-03-29 | 海南省农业科学院植物保护研究所(海南省农业科学院农产品质量安全与标准研究中心) | Bacillus amyloliquefaciens Yb-2 and separation method and application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104762223B (en) * | 2014-11-06 | 2018-05-08 | 河北省科学院生物研究所 | A kind of bacillus amyloliquefaciens BA-KA3 and its application |
| CN109504639A (en) * | 2018-12-29 | 2019-03-22 | 贵州大学 | Bacillus amyloliquefaciens GUMT319 and its application |
| CN109504638A (en) * | 2018-12-29 | 2019-03-22 | 贵州大学 | Bacillus amyloliquefaciens GUAL210 and its application |
| CN109762759A (en) * | 2018-12-29 | 2019-05-17 | 贵州大学 | Bacillus amyloliquefaciens GUAL203 and its application |
| CN109852564A (en) * | 2019-03-08 | 2019-06-07 | 贵州省烟草公司黔西南州公司 | Bacillus amyloliquefaciens 327 and its application |
-
2019
- 2019-06-19 CN CN201910531471.8A patent/CN110257286A/en not_active Withdrawn
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104762223B (en) * | 2014-11-06 | 2018-05-08 | 河北省科学院生物研究所 | A kind of bacillus amyloliquefaciens BA-KA3 and its application |
| CN109504639A (en) * | 2018-12-29 | 2019-03-22 | 贵州大学 | Bacillus amyloliquefaciens GUMT319 and its application |
| CN109504638A (en) * | 2018-12-29 | 2019-03-22 | 贵州大学 | Bacillus amyloliquefaciens GUAL210 and its application |
| CN109762759A (en) * | 2018-12-29 | 2019-05-17 | 贵州大学 | Bacillus amyloliquefaciens GUAL203 and its application |
| CN109852564A (en) * | 2019-03-08 | 2019-06-07 | 贵州省烟草公司黔西南州公司 | Bacillus amyloliquefaciens 327 and its application |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110747208A (en) * | 2019-11-28 | 2020-02-04 | 海南大学 | Cassava nitrate reductase gene and construction and disease-resistant application of overexpression vector thereof |
| CN110747208B (en) * | 2019-11-28 | 2022-04-22 | 海南大学 | Cassava nitrate reductase gene and construction and disease-resistant application of overexpression vector thereof |
| CN114250171A (en) * | 2021-12-08 | 2022-03-29 | 海南省农业科学院植物保护研究所(海南省农业科学院农产品质量安全与标准研究中心) | Bacillus amyloliquefaciens Yb-2 and separation method and application thereof |
| CN114250171B (en) * | 2021-12-08 | 2023-07-18 | 海南省农业科学院植物保护研究所(海南省农业科学院农产品质量安全与标准研究中心) | Bacillus amyloliquefaciens Yb-2 and separation method and application thereof |
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Application publication date: 20190920 |