CN104560837B - A kind of bacillus amyloliquefaciens and its application - Google Patents
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Abstract
The present invention relates to functional microorganism triage techniques field, a kind of Bacillus amyloliquefaciens strain is specifically provided, deposit number is CCTCC NO：M2015075.The bacillus amyloliquefaciens can significantly inhibit the growth and breeding of the pathogens such as the single softgel shell bacterium of garment or robe made of feathers grass, botrytis, Fusarium oxysporum and Rhizoctonia solani Kuhn, effectively prevent and treat the plant diseases such as powdery mildew of strawberry, gray mold of cucumber, effect is notable.Bacillus amyloliquefaciens of the present invention can be applied to the prevention and treatment of plant disease separately as biocontrol agent, can also have bacillus, mould of bacteriostasis etc. to be combined into complex microorganism preparations with other, efficiency of preventing and treating to plant diseases such as powdery mildew, gray mold, brown spot, fruit rot reaches 85% 90%, has a extensive future.
Description
Technical field
The present invention relates to functional microorganism triage techniques field, and in particular to a kind of bacillus amyloliquefaciens and its in plant
Application in disease control.
Background technology
It is on the increase with floristic, the continuous variation of plant culture mode, plant disease is on the rise, and passes
The chemical prevention and control method of system has many drawbacks, such as：A large amount of use of chemical pesticide causes serious environmental pollution, agriculture
The residues of pesticides of product are also increasingly severe, and the germ resistance to the action of a drug produces speed, and also a series of increasingly faster etc. lived to people is harmful to
Influence.In low-carbon economy, environmental protection economical society is advocated now, seek that human lives can be protected, but it is fertile new
Technology extremely adapts to the development of today's society.
Biological control method is sustainable control and the Unpolluting management that harmful organism is reached using bio-diversity, is met
The development and requirement of today's society, have broad application prospects.Wherein, microbial control mainly uses beneficial micro- life
Thing, by interbiotic Competition, antibiosis, parasitization, bacteriolysis and induction of resistance etc., to suppress some cause of diseases
The survival and activity of thing, i.e., using in the ecosystem it is various it is biological between interdependence, the ecological phenomenon that mutually restricts and certain
A little biological characteristicses, so that an a kind of or class is biological or its Product inhibiton is another or another kind of biology.In short, worm is controlled with bacterium,
Cured the disease with bacterium, it is advantageous in that aboundresources, selective strong, noresidue, pollution-free, cost, low and anti-simultaneous control increase production increasing
Receive, keep ecological balance, play long-acting effect.So exploitation microbial pesticide has broad application prospects.(Zhang Li, Zhang Shu
Pretty young woman；2010)
It has now been found that, the microorganism available for biological control mainly includes fungi, bacterium, actinomyces and saccharomycete etc.
Microbial population.But, reaction of the Biocontrol microorganism to agricultural chemicals is sensitive, and species is relatively simple, and antimicrobial spectrum is narrower, and on plant
It is more difficult to colonize, and biocontrol effect is unstable, all limits its extensive use in plant field.Therefore, further exploitation wide spectrum resists
The stable microbial resources of bacterium, biocontrol effect are extremely urgent.
The content of the invention
Application it is an object of the invention to provide a bacillus amyloliquefaciens and its in control of plant disease.The solution
Bacillus amyloliquefacienses are screened from rhizosphere of strawberry soil, to the single softgel shell bacterium of garment or robe made of feathers grass, Fusarium oxysporum, Rhizoctonia solani Kuhn, grape spore
The pathogens such as bacterium have very strong rejection ability, can effectively prevent and treat powdery mildew, droop, brown spot, fruit rot, gray mold etc. often
See plant disease, effect is notable.
One aspect of the present invention provides a kind of bacillus amyloliquefaciens VLD-3 (Bacillus amyloliquefaciens
VLD-3), the China typical culture collection center for being preserved in Wuhan, China Wuhan University on January 30th, 2015, its preservation
Number be CCTCC NO：M2015075.
Another aspect of the present invention provides a kind of microorganism formulation, includes above-mentioned bacillus amyloliquefaciens VLD-3.
The microorganism formulation also includes bacillus subtilis, bacillus megaterium, bacillus pumilus, strong gemma bar
Any one or two or more combination in bacterium, bacillus cereus, bacillusmusilaginosiengineering and aspergillus niger.
Bacillus amyloliquefaciens VLD-3 viable bacteria amount is at least 10 in the microorganism formulation7CFU/ml。
Present invention also offers application of the mentioned microorganism preparation in control of plant disease.
The plant disease includes any one in powdery mildew, gray mold, droop, brown spot and fruit rot.
Beneficial effect
The bacillus amyloliquefaciens VLD-3 that the present invention is screened can significantly inhibit the single softgel shell bacterium of garment or robe made of feathers grass, botrytis, point
The growth and breeding of the pathogen such as fusarium oxysporum and Rhizoctonia solani Kuhn.The bacterial strain can effectively prevent the generation of powdery mildew of strawberry, real
The disease index of group strawberry is tested well below control group, and when bacillus amyloliquefaciens VLD-3 bacterium solutions are diluted to 700 times, it is real
The disease index for testing group strawberry is still below 8%, prevents and treats efficiency high up to 91.8%；In addition, the bacterial strain is heavier to powdery mildew epidemic situation
Strawberry and the heavier cucumber of gray mold epidemic situation have a significant therapeutic effect, the bacillus amyloliquefaciens VLD- after 50 times of dilutions
3 bacterium solutions can make the powdery mildew disease index of strawberry be reduced to 14.2%, and the gray mold disease index of cucumber is reduced to 13.2%, and
The disease index of control group is generally higher than 96%, achieves unexpected technique effect.Solution starch gemma of the present invention
Bacillus VLD-3 can be applied to the prevention and treatment of plant disease separately as biocontrol agent, can also have antibacterial work with other
Bacillus, mould etc. are combined into composite bacteria agent, to plant diseases such as plant powdery mildew, gray mold, brown spot, fruit rot
Efficiency of preventing and treating reach 85%-90%, have a extensive future.
Embodiment
The present invention is expanded on further with reference to specific embodiment.For specific method or material used in embodiment
Material, those skilled in the art can be carried out conventional replacement according to existing technology and selected on the basis of the technology of the present invention thinking
Select, be not limited solely to the specific record of the embodiment of the present invention.
Equipment and reagent selected by the present invention can selected from it is commercially available any one.
The separation screening of the bacterial strain of embodiment 1 and identification
1. bacillus primary dcreening operation
The strawberry rhizosphere soil 10g of Qingdao City Laoshan District Bei Zhai planting bases is gathered, is put into and fills 40ml aqua sterilisas
Conical flask in；After concussion is mixed, supernatant is transferred to new sterilizing test tubes；By test tube through 80 DEG C of water-bath 20min processing,
Kill non-spore vegetative cells；Above-mentioned solution is diluted after 20 times, 100 μ l is drawn and is coated on nutrient broth medium, 37 DEG C
Cultivate 12h.After the different bacterium colony of form is purified into single bacterium colony through plate streaking, nutrient broth inclined-plane, 37 DEG C of cultures are moved into
12h, 4 DEG C of refrigerators are saved backup.
Using above-mentioned method for screening and separating, 23 bacillus are obtained altogether.
2. bacteriostatic experiment
(1) prepared by bacillus bacterium solution：
23 isolated bacillus are inoculated in nutrient broth medium respectively, 37 DEG C, 200rpm cultures
12h；(2) prepared by pathogen：
The fungus block (Φ 7mm) of Fusarium oxysporum or Rhizoctonia solani Kuhn (being given by Qingdao Agricultural University) is connected to PDA solids
Culture medium central, 30 DEG C of culture 7d；
(3) flat board antagonism method primary dcreening operation：
In the center inoculation Fusarium oxysporum of PDA solid mediums or Rhizoctonia solani Kuhn fungus block (Φ 7mm)；Then, away from
Culture dish center 25mm symmetrical both sides point connects the above-mentioned bacillus bacterium solution screened, per bacillus set 3 it is parallel.Will
The flat board of pathogen and bacillus is put into after 30 DEG C of incubators, 72h it has been observed that having the week of 8 bacillus in inoculation
Enclose to form obvious antibacterial band, VLD-1, VLD-2, VLD-3 ... ..., VLD-8 are named as respectively.
(4) diffusion method secondary screening is punched：
PDA culture medium temperature after sterilizing is maintained at 55 DEG C, pathogen (sharp spore reaping hook is then gently rinsed with sterilized water
Bacterium or Rhizoctonia solani Kuhn) inclined-plane obtains pathogen spore suspension, and take in appropriate addition PDA culture medium, plate is down flat after mixing.Treat
After solidification, that is, it is about 10 that cause of disease bacterial content, which is made,7Cfu/ml PDA solid mediums.With Oxford cup away from culture dish center
25mm symmetrical both sides respectively make a hole (Φ 7mm), and add 150 μ l bacillus (VLD-1, VLD-2, VLD- to be measured
3 ... ..., VLD-8) bacterium solution in hole, set per bacillus 3 it is parallel.Flat board is put into after 30 DEG C of incubators, 72h and surveyed
Measure the diameter of inhibition zone.As a result show, the antibacterial circle diameter for adding the flat board of bacillus VLD-3 bacterium solutions is maximum, so as to
Illustrate, VLD-3 fungistatic effect is most strong in 8 bacillus that the present invention is screened.
3. bacterial strain is identified
Bacillus VLD-3 is subjected to molecular biology identification, its 16s rRNA sequence is measured, and pass through NCBI BLAST
Compare, it is bacillus amyloliquefaciens (Bacillus amyloliquefaciens) finally to confirm the bacterial strain, is named as solution
Bacillus amyloliquefacienses VLD-3 (Bacillus amyloliquefaciens VLD-3), and on January 30th, 2015 is preserved in
The China typical culture collection center of Wuhan Wuhan University of state, deposit number is CCTCC NO：M2015075..
The bacillus amyloliquefaciens VLD-3 bacteriostatic experiments of embodiment 2
By the single softgel shell bacterium of garment or robe made of feathers grass, botrytis, 4 kinds of pathogen fungus blocks of Fusarium oxysporum and Rhizoctonia solani Kuhn (Φ 10mm)
It is respectively placed in PDA solid cultures plate (Φ 90mm) center；With the side of a diameter of 60mm sterile blank plate (Φ 60mm)
It is 10 that edge, which dips viable bacteria concentration,9After cfu/ml bacillus amyloliquefaciens VLD-3 bacterium solutions, in above-mentioned PDA solid cultures plate
Cause of disease fungus block around carry out annular inoculation.Blank control group is then that annular inoculation is carried out with sterilized water.PDA after inoculation is consolidated
Body culture dish is placed in 30 DEG C of incubators, respectively when cultivating 66h, 90h, 120h and 190h, measures the diameter of cause of disease fungus block,
Bacteriostasis rate is calculated using below equation, concrete outcome is shown in Table 1.
Bacteriostasis rate (%)=(1- (treatment group pathogen spot diameter/control group pathogen spot diameter)) × 100
Fungistatic effects of the bacillus amyloliquefaciens VLD-3 of table 1 to pathogen
Can be seen that the bacillus amyloliquefaciens VLD-3 of the invention screened from the experimental result of table 1 can significantly inhibit plumage
The single softgel shell bacterium of clothing grass, botrytis, the breeding of Fusarium oxysporum and Rhizoctonia solani Kuhn, and bacteriostasis rate reaches substantially when cultivating 120h
To peak value, bacteriostasis rate is generally higher than 55%, and effect is notable.
Preventive effects of the bacillus amyloliquefaciens VLD-3 of embodiment 3 to powdery mildew of strawberry
1st, prepared by bacterium solution
Bacillus amyloliquefaciens VLD-3 is inoculated in liquid nutrient broth culture medium, 37 DEG C, 200rpm culture 12h are obtained
Viable bacteria content is 108-109Cfu/ml bacterium solution.
2nd, place is tested：Qingdao City Chengyang starts the strawberry greenhouse of planting base；
3rd, application method
1) experimental group：Three treatment groups, each ridge strawberry for the treatment of group 10, sets up guarantor between each treatment group by totally 1800
Shield row；
Come into bloom in strawberry, bacillus amyloliquefaciens VLD-3 bacterium solutions are diluted with water 300, after 500,700 times, respectively
It is uniform to be sprayed at Strawberry Leaves anyway on face and stem；Sprinkling interval 7 days, is used in conjunction 3 times every time；Last time has sprayed bacterium solution 10 days
Afterwards, the incidence of powdery mildew of strawberry is checked.
2) control group：Using the nutrient broth medium without bacterium, other operations and experimental group all same.
4th, interpretation of result
Disease scale standard：0 grade of disease-free spot；1 grade：Lesion area accounts for less than the 1/4 of leaf area；2 grades：Lesion area accounts for leaf
The 1/4-2/4 of area；3 grades：Lesion area accounts for the 2/4-3/4 of leaf area；4 grades：Lesion area accounts for more than the 3/4 of leaf area.
Disease index=[∑ (the sick level value × level morbidity number of sheets)/(the sick level value of investigation total number of sheets × highest)] × 100%
Prevent and treat calculation formula=[1- (disease index of disease index/control of processing)] × 100% of efficiency.
Experimental data is analyzed with SPSS statistical softwares, as a result as shown in table 2：
Preventive effects of the bacillus amyloliquefaciens VLD-3 of table 2 to powdery mildew of strawberry
Can be seen that the bacillus amyloliquefaciens VLD-3 of the invention screened from the data of table 2 can effectively suppress pathogen
The growth and breeding of (the mainly single softgel shell bacterium of garment or robe made of feathers grass), prevents the generation of powdery mildew of strawberry, the disease index of experimental group strawberry is remote
Far below control group, and when bacillus amyloliquefaciens VLD-3 bacterium solutions are diluted to 700 times, the disease index of experimental group strawberry is still
Less than 8%, efficiency high is prevented and treated up to 91.8%, unexpected preventive effect is achieved.
Therapeutic effects of the bacillus amyloliquefaciens VLD-3 of embodiment 4 to powdery mildew of strawberry
1st, prepared by bacterium solution
Bacillus amyloliquefaciens VLD-3 is inoculated in liquid nutrient broth culture medium, 37 DEG C, 200rpm culture 12h are obtained
Viable bacteria content is 108-109Cfu/ml bacterium solution.
2nd, place is tested：The strawberry greenhouse of Qingdao City Jimo crossdrift town planting base；
3rd, application method
1) experimental group：Three treatment groups, each ridge strawberry for the treatment of group 10, sets up guarantor between each treatment group by totally 1800
Shield row；
When powdery mildew of strawberry epidemic situation is heavier (every plant sick leaf rate up to more than 50%), bacillus amyloliquefaciens VLD-3 is added
Water dilution 50,100, after 200 times, Strawberry Leaves are uniformly sprayed at respectively anyway on face and stem, are 1L per ridge fountain height；Spray every time
Spill interval 5 days, be used in conjunction 5 times；After last time has sprayed bacterium solution 7 days, the treatment of powdery mildew of strawberry is checked.
2) control group：Using the nutrient broth medium without bacterium, other operations and experimental group all same.
According to interpretation of result method described in embodiment 3, experimental data is analyzed with SPSS statistical softwares, as a result such as table
Shown in 3：
Therapeutic effects of the bacillus amyloliquefaciens VLD-3 of table 3 to powdery mildew of strawberry
Can be seen that the bacillus amyloliquefaciens VLD-3 of the invention screened from the data of table 3 can effectively kill pathogen
(the mainly single softgel shell bacterium of garment or robe made of feathers grass), the strawberry heavier to powdery mildew epidemic situation has significant therapeutic effect, the solution after 50 times of dilutions
Bacillus amyloliquefacienses VLD-3 bacterium solutions can make the powdery mildew disease index of strawberry be reduced to less than 15%, and the state of an illness of control group refers to
Number is up to 96.6%, achieves unexpected technique effect.
The therapeutic effect of the gray mold of cucumber of VLD-3 pairs of 5 bacillus amyloliquefaciens of embodiment
1st, prepared by bacterium solution
Bacillus amyloliquefaciens VLD-3 is inoculated in liquid nutrient broth culture medium, 37 DEG C, 200rpm culture 12h are obtained
Viable bacteria content is 108-109Cfu/ml bacterium solution.
2nd, place is tested：The cucumber booth in Yantai City Haiyang vegetable cultivation base；
3rd, application method
1) experimental group：Three treatment groups, each ridge strawberry for the treatment of group 10, sets up protection between each treatment group by totally 360
OK；
When gray mold of cucumber epidemic situation is heavier (every plant sick leaf rate up to more than 50%), bacillus amyloliquefaciens VLD-3 is added
Water dilution 50,100, after 200 times, cucumber leaves are uniformly sprayed at respectively anyway on face and melon stem, are 2L per ridge fountain height；Every time
Sprinkling interval 5 days, is used in conjunction 5 times；After last time has sprayed bacterium solution 6 days, the treatment of powdery mildew of strawberry is checked.
2) control group：Using the nutrient broth medium without bacterium, other operations and experimental group all same.
According to interpretation of result method described in embodiment 3, experimental data is analyzed with SPSS statistical softwares, as a result such as table
Shown in 4：
Therapeutic effects of the bacillus amyloliquefaciens VLD-3 of table 4 to gray mold of cucumber
Can be seen that the bacillus amyloliquefaciens VLD-3 of the invention screened from the data of table 4 can effectively kill pathogen
(mainly botrytis), the cucumber heavier to gray mold epidemic situation has significant therapeutic effect, the solution starch bud after 50 times of dilutions
Spore bacillus VLD-3 bacterium solutions can make the gray mold disease index of cucumber be reduced to 13.2%, prevent and treat efficiency high and reach 86.5%, treatment
Effect is obvious.
Further, applicant is had found by many experiments, and in addition to plant powdery mildew and gray mold, the present invention is screened
Bacillus amyloliquefaciens VLD-3 also to by sickle-like bacteria and the microbial plant droop of miliary damping-off, brown spot and fruit rot
Deng there is obvious antagonism, 85%-95% can be reached by preventing and treating efficiency.
Bacillus amyloliquefaciens VLD-3 of the present invention can separately as biocontrol agent be applied to plant disease prevention and
Treatment, also can with bacillus subtilis (Bacillus subtilis), bacillus megaterium (Bacillus megaterium),
Bacillus pumilus (Bacillus pumilus), bacillus firmus (Bacillus firmus), bacillus cereus
(Bacillus cereus), bacillusmusilaginosiengineering (Bacillus mucilaginosus) and aspergillus niger (Aspergillus
Niger any one or two or more the combination in), prepares complex microorganism preparations.The complex microorganism system
Agent reaches 85%-90% to the efficiency of preventing and treating of the plant diseases such as plant powdery mildew, gray mold, droop, brown spot, fruit rot,
Have a extensive future.
Claims (6)
1. a kind of bacillus amyloliquefaciens, it is characterised in that the preserving number of described bacillus amyloliquefaciens is CCTCC NO：
M2015075。
2. a kind of microorganism formulation, it is characterised in that described microorganism formulation includes the solution starch gemma described in claim 1
Bacillus.
3. microorganism formulation as claimed in claim 2, it is characterised in that described microorganism formulation includes bacillus subtilis
Bacterium, bacillus megaterium, bacillus pumilus, bacillus firmus, bacillus cereus, bacillusmusilaginosiengineering and aspergillus niger
In any one or two or more combination.
4. microorganism formulation as claimed in claim 2 or claim 3, it is characterised in that claim 1 institute in described microorganism formulation
The viable bacteria amount for the bacillus amyloliquefaciens stated is at least 107 CFU/ml。
5. application of the microorganism formulation described in Claims 2 or 3 in control of plant disease.
6. application as claimed in claim 5, it is characterised in that described plant disease be powdery mildew, gray mold, droop,
Any one in brown spot and fruit rot.
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| CN105028483B (en) * | 2015-07-27 | 2018-01-30 | 河北省农林科学院植物保护研究所 | Composite bactericide and its application containing bacillus subtilis BAB 1 and Boscalid |
| CN105532735A (en) * | 2015-11-21 | 2016-05-04 | 安徽省农业科学院植物保护与农产品质量安全研究所 | Bactericidal and insecticidal composition containing Bacillus amyloliquefaciens and matrine |
| CN106701618B (en) * | 2016-12-09 | 2020-04-14 | 武汉科诺生物科技股份有限公司 | Bacillus amyloliquefaciens capable of resisting powdery mildew of melon crops and producing spores at high yield |
| CN106967632A (en) * | 2017-02-23 | 2017-07-21 | 南京农业大学 | It is a kind of efficiently to prevent and treat various crop gray mold and the microorganism mixture of capsicum resistance be lifted |
| CN107043719B (en) * | 2017-03-16 | 2019-12-10 | 河北省农林科学院植物保护研究所 | bacillus amyloliquefaciens HMB28353 and application thereof |
| CN107043720B (en) * | 2017-03-16 | 2019-12-10 | 河北省农林科学院植物保护研究所 | Bacillus amyloliquefaciens HMB28388 and application thereof |
| CN107047618A (en) * | 2017-05-02 | 2017-08-18 | 陕西康禾立丰生物科技药业有限公司 | A kind of bactericidal composition containing bacillus amyloliquefaciens |
| CN108991019A (en) * | 2018-07-06 | 2018-12-14 | 东北农业大学 | The biocontrol agent composition for preventing and treating graw mold of tomato and leaf mold |
| CN108977374A (en) * | 2018-07-06 | 2018-12-11 | 东北农业大学 | The compounding antimicrobial composition for preventing and treating graw mold of tomato and leaf mold |
| CN108998395B (en) * | 2018-08-24 | 2021-05-18 | 浙江省农业科学院 | Bacillus amyloliquefaciens and application thereof |
| CN109294947A (en) * | 2018-10-08 | 2019-02-01 | 山东蔚蓝绿源生物科技有限公司 | A kind of separation screening and zymotechnique of the bacillus amyloliquefaciens for powdery mildew |
| CN109355233B (en) * | 2018-12-04 | 2021-04-09 | 沈阳化工研究院有限公司 | Bacillus amyloliquefaciens and application thereof |
| CN109554319B (en) * | 2019-01-31 | 2021-03-16 | 中国科学院微生物研究所 | Bacillus strain and application thereof |
| CN110724648B (en) * | 2019-08-30 | 2021-07-20 | 山东蔚蓝生物科技有限公司 | Bacillus amyloliquefaciens strain and application thereof |
| CN114437965B (en) * | 2021-12-31 | 2022-12-20 | 河北科技师范学院 | Peanut rhizosphere microbial strain and method for improving and repairing soil by using biological bacterial fertilizer thereof |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1952116A (en) * | 2006-04-18 | 2007-04-25 | 兰州大学 | Bacillusamyloliquefaciens strain and application thereof |
| CN101555459A (en) * | 2009-03-31 | 2009-10-14 | 江苏丘陵地区镇江农业科学研究所 | Strain SB177 for controlling strawberry replant diseases and preparation thereof |
| CN102816713A (en) * | 2012-06-12 | 2012-12-12 | 江苏农林职业技术学院 | Bacillus amyloliquefaciens and its application |
| CN103087951A (en) * | 2013-01-14 | 2013-05-08 | 北京市农林科学院 | Bacillus amyloliquefaciens and application thereof |
| CN103173397A (en) * | 2013-04-02 | 2013-06-26 | 山东省农业科学院农产品研究所 | Broad-spectrum antibacterial bacillus amyloliquefaciens strain and application thereof |
| CN103602606A (en) * | 2013-06-24 | 2014-02-26 | 江苏农林职业技术学院 | Bacillus amyloliquefaciens and application thereof in preventing and treating of plant fruit rot diseases |
| CN103642727A (en) * | 2013-12-02 | 2014-03-19 | 北京市农林科学院 | Bacillus amyloliquefaciens inhibiting fungi and application thereof |
-
2015
- 2015-02-02 CN CN201510054680.XA patent/CN104560837B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1952116A (en) * | 2006-04-18 | 2007-04-25 | 兰州大学 | Bacillusamyloliquefaciens strain and application thereof |
| CN101555459A (en) * | 2009-03-31 | 2009-10-14 | 江苏丘陵地区镇江农业科学研究所 | Strain SB177 for controlling strawberry replant diseases and preparation thereof |
| CN102816713A (en) * | 2012-06-12 | 2012-12-12 | 江苏农林职业技术学院 | Bacillus amyloliquefaciens and its application |
| CN103087951A (en) * | 2013-01-14 | 2013-05-08 | 北京市农林科学院 | Bacillus amyloliquefaciens and application thereof |
| CN103173397A (en) * | 2013-04-02 | 2013-06-26 | 山东省农业科学院农产品研究所 | Broad-spectrum antibacterial bacillus amyloliquefaciens strain and application thereof |
| CN103602606A (en) * | 2013-06-24 | 2014-02-26 | 江苏农林职业技术学院 | Bacillus amyloliquefaciens and application thereof in preventing and treating of plant fruit rot diseases |
| CN103642727A (en) * | 2013-12-02 | 2014-03-19 | 北京市农林科学院 | Bacillus amyloliquefaciens inhibiting fungi and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| 一株新的拮抗细菌SL19及其抑菌活性物质;刘莲娜等;《微生物学通报》;20110820;第38卷(第8期);1199-1206 * |
| 大棚草莓白粉病及其防治;黄根元等;《植物保护》;20010630;第27卷(第3期);46-47 * |
| 解淀粉芽孢杆菌LJ1 诱导黄瓜抗白粉病的研究;谷医林等;《农药学学报》;20131231;第15卷(第3期);293-298 * |
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