CN109022332A - One plant of biological and ecological methods to prevent plant disease, pests, and erosion Bei Laisi bacillus and its application in prevention and treatment cucumber powdery mildew's pathogen - Google Patents
One plant of biological and ecological methods to prevent plant disease, pests, and erosion Bei Laisi bacillus and its application in prevention and treatment cucumber powdery mildew's pathogen Download PDFInfo
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- CN109022332A CN109022332A CN201811068345.5A CN201811068345A CN109022332A CN 109022332 A CN109022332 A CN 109022332A CN 201811068345 A CN201811068345 A CN 201811068345A CN 109022332 A CN109022332 A CN 109022332A
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Abstract
It is TB1501 the present invention relates to one plant of Bei Laisi bacillus strain (Bacillus velezensis) Strain Designation, which is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation registration number are as follows: CGMCC No.16069；The preservation time are as follows: on July 6th, 2018.Bacterial strain significant effect in terms of preventing and treating cucumber powdery mildew's pathogen.Typical efficiency test is as the result is shown: TB1501 bacterial strain plays good control efficiency during prevent and treat cucumber powdery mildew's pathogen, and preventive effect is up to 85% or more, almost suitable with pharmaceutical preparations Flutriafol better than chemical agent Tebuconazole.
Description
Technical field
The present invention relates to plant disease technical fields, and in particular to one plant of biological and ecological methods to prevent plant disease, pests, and erosion Bei Laisi bacillus TB1501 and
Its application in prevention and treatment cucumber powdery mildew's pathogen.
Background technique
Powdery mildew is commonly called as " white hair disease ", is one of most important plant disease in current China, in addition to coniferous tree, each
Kind of cereal crops, industrial crops, gourd, fruit and vegetable, ornamental plant have generation, and have a very wide distribution, and each growing area is all at home
There is generation.In the north of China greenhouse ground family crop using cucumber as representative, powdery mildew harm is particularly acute, and is easily caused larger
Economic loss (up to 50% or more).Rapidly due to the disease short incubation period, morbidity, it must frequently be administered in production to carry out prevention and control, with
For powdery mildew of cucumber, it must spray at the season in spring and autumn of onset peak usual 7-10 days primary, to cause serious pesticide
Residue problem not only directly results in serious economic loss and threatens people's health, in some instances it may even be possible to serious society be caused to be asked
Topic.The strategy for controlling the quantity of phytopathogen using beneficial microbe in nature or its metabolite or mitigating its harm claims
For biocontrol of plant disease.Bacillus has resistance strong, and breeding is fast, in the strong characteristic of plant rhizosphere colonization ability, is
The important sources of microbial bactericide.Currently, Bacillus subtillis, bacillus licheniformis, bacillus amyloliquefaciens and waxy
Bacillus etc. is the main source of China's microbial bactericide.Under the overall background of " medicine fertilizer is double to be subtracted " that country advocates, safety,
Environmental-friendly biological pesticide is alternative or partially substitutes the use of chemical agent, to reduce the usage amount of chemical pesticide.Cause
This reinforces the exploitation of biological pesticide and study on mechanism is the urgent task of current plant protection field researcher, has important
Science and Social Ecology meaning.
Summary of the invention
To achieve the above object, the invention discloses one plant of Bei Laisi bacillus strain (Bacillus velezensis)
Strain Designation is TB1501, it is characterised in that the bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms
Center, preservation registration number are as follows: CGMCC No. 16069；The preservation time are as follows: on July 6th, 2018.
Bei Laisi bacillus TB1501 is screened by biocontrol of plant disease research department of Tianjin Plant Protection Institute
Obtain, TB1501 by Institute of Microorganism, Academia Sinica be accredited as one plant of Bei Laisi bacillus (Bacillus velezensis), and it is stored in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation registration number are as follows:
CGMCC No. 16069；The preservation time are as follows: on July 6th, 2018.China Committee for Culture Collection of Microorganisms's commonly micro- life
Object centre address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica, postcode: 100101.
The strain of Bei Laisi bacillus (Bacillus velezensis) be named as TB1501 physio-biochemical characteristics it is as follows:
Bei Laisi bacillus TB1501 is in solid LB media (beef extract powder 5g/L, sodium chloride 10g/L, tryptone
10g/L, 15 ~ 20 g/L of agar) on bacterium colony be creamy white, shape is approximate circle, and edge is irregular, and there is fold on surface, and thallus is in
Rod-shaped, gemma is closely oval.During practical application, it is contemplated that it may need the reasons such as transport, it is necessary to be expanded
The form of microbial bacterial agent is made to expand its application range in big culture.
Thus the microbial bacteria of the metabolite containing bacterial strain as described above and/or the bacterial strain is also claimed in the present invention
Agent.Preferably, microbial bacterial agent as described above, the microbial bacterial agent are liquid bacterial agent.And in the microbial inoculum viable bacteria and/
Or quantity >=50 of living spores~6,000,000,000/g.
The present invention further discloses the preparation method of the microbial bacterial agent containing Bei Laisi Bacillus strain, features
It is to carry out by following step:
(1) activate TB1501 bacterial strain: one ring TB1501 bacterium colony of picking is seeded in 5mL LB liquid medium, 37 DEG C, 160rpm
Shake culture overnight, is made TB1501 seed liquor；
(2) solid LB media is prepared: 10g/L tryptone, 10g/L sodium chloride, 5g/L beef extract powder, 15-20g/L fine jade
Cosmetics, deionized water dissolving, 121 DEG C, moist heat sterilization 30min；
(3) LB liquid medium is prepared: 10g/L tryptone, 10g/L sodium chloride, 5g/L beef extract powder, deionization are water-soluble
Solution, 121 DEG C, moist heat sterilization 30min；
(4) seed liquor that will be prepared, 1/1000 is inoculated in 100 milliliters of liquid LB culture mediums, 160rpm, cultivates 48-72h,
The TB1501 fermentation liquid as prepared.
The present invention further disclose the strain of Bei Laisi bacillus (Bacillus velezensis) it is named as TB1501
Application in prevention and treatment cucumber powdery mildew's pathogen.The bacterium can preferably prevent and treat cucumber powdery mildew's pathogen, have good Biocontrol Potential.
Fig. 1 is colonial morphology of the TB1501 on solid LB media, and Fig. 2 is the thalli morphology that TB1501 is observed under oil mirror.Allusion quotation
The efficiency test of type is as the result is shown: TB1501 bacterial strain plays good prevention and treatment effect during preventing and treating cucumber powdery mildew's pathogen
Fruit, preventive effect are up to 85% or more, are better than chemical agent Tebuconazole, almost suitable with pharmaceutical preparations Flutriafol.
The main physical and chemical of the strain preparation of Bei Laisi bacillus prepared by the present embodiment, as follows:
Bei Laisi bacillus of the present invention strain (Bacillus velezensis) it is named as the analysis of effective component method of TB1501
Are as follows:
Using bacterium colony colony counting method, it will quantitatively be coated on training LB solid after the dilution of bacteria suspension certain multiple and support in base, to shellfish Lay
The strain of this bacillus grows bacterium colony in media surface, Bei Laisi bacillus strain clump count is counted, to measure the unit of sample
Weight living spores number.
The toxicology of Bei Laisi bacillus strain of the present invention:
According to China's pesticide toxicity grading criteria, 100,000,000,000 Bei Laisi bacillus strain preparations belong to less toxic fungicide.To rabbit eyes
There is slight stimulation, is weak sensitizer to cavy to rabbit no skin irritation.
The invention discloses one plant of Bei Laisi bacillus (Bacillus velezensis) bacterial strain and prior art phase
It is had the active effect that than possessed
(1) Bei Laisi bacillus TB1501 bacterial strain belongs to bacillus, and liquid bacterial agent preparation method is simple.
(2) Bei Laisi bacillus TB1501 bacterial strain application and popularization value in terms of powdery mildew of cucumber prevention and treatment is higher.
(3) application of Bei Laisi bacillus TB1501 bacterial strain can reduce the use of pesticide, in protection and rehabilitating soil
There is positive meaning in terms of micro-ecological environment.
Detailed description of the invention
Fig. 1 is colonial morphology of the TB1501 on solid LB media；
Fig. 2 is the thalli morphology that TB1501 is observed under oil mirror；
Fig. 3 is TB1501 and Flutriafol, Tebuconazole preventive effect compare.
Specific embodiment
The present invention is described below by specific embodiment.Unless stated otherwise, technological means used in the present invention
It is method known in those skilled in the art.In addition, embodiment is interpreted as illustrative, it is not intended to limit the present invention
Range, the spirit and scope of the invention are limited only by the claims that follow.To those skilled in the art, without departing substantially from this
Under the premise of invention spirit and scope, to the various changes or change of material component and dosage progress in these embodiments
It belongs to the scope of protection of the present invention.The raw materials used in the present invention and reagent are commercially available.
Embodiment 1
(1) Bei Laisi bacillus TB1501 is screened by biocontrol of plant disease research department of Tianjin Plant Protection Institute
Arrive, TB1501 by Institute of Microorganism, Academia Sinica be accredited as one plant of Bei Laisi bacillus (Bacillus velezensis), and it is stored in China Committee for Culture Collection of Microorganisms's common micro-organisms center, the preservation time are as follows:
On July 6th, 2018；Preservation registration number are as follows: CGMCC No. 16069.
(2) screening technique of Bei Laisi bacillus TB1501 bacterial strain:
TB1501 bacterial strain is isolated from vegetables rhizosphere soil, sampling spot: Tianjin Wuqing District.
(1) 1 gram of soil is weighed, is put into 50 mL triangular flasks, sterile tap water 9 mL, 100rpm is added, concussion mixes
Soil bacteria suspension is made in 30min；
(2) soil sample bacteria suspension is put into 85 DEG C of water-bath 15min, rear Aspirate supernatant is diluted 100 times, 1000 times；
(3) by 100 times, 1000 times of supernatant dilution applying solid LB plates (10g/L tryptone, 10g/L sodium chloride, 5g/L
Beef extract powder, 15g/L agar powder), 37 DEG C are incubated overnight；
(4) Bacillus colonies on plate are chosen, save storage in case raw survey uses.
Powdery mildew of cucumber biocontrol microorganisms primary dcreening operation:
Laboratory has the biocontrol microorganisms library (these biocontrol microorganisms are used as experiment, externally can be with obtainable post-free) of oneself, storage capacity 5000
Strain prevents and treats preferable biocontrol microorganisms to screen cucumber powdery mildew's pathogen.
(1) one ring biocontrol microorganisms of picking access 5 mL LB liquid mediums (10g/L tryptone, 10g/L sodium chloride, 5g/L
Beef extract powder), 37 DEG C, 150rpm is incubated overnight and biological and ecological methods to prevent plant disease, pests, and erosion fermented liquid is made；
(2) growth conditions are chosen, health cucumber leaf of the same size sprays × 100 biological and ecological methods to prevent plant disease, pests, and erosion fermented liquids, inoculation is yellow two days later
Melon Powdery Mildew (preservation of this laboratory) spore suspension (105A spore/mL), it is investigated after 10 days.Investigation result is shown
TB1501 bacterial strain prevention and treatment powdery mildew of cucumber effect is best, reaches 75%, saves TB1501 as scientific research bacterial strain.
Embodiment 2
The preparation of TB1501 fermentation liquid:
(1) TB1501 bacterial strain is activated.One ring TB1501 bacterium colony of picking, is seeded in 5mL LB liquid medium, 37 DEG C, 160rpm
Shake culture overnight, is made TB1501 seed liquor.
(2) solid LB media is prepared: 10g/L tryptone, 10g/L sodium chloride, 5g/L beef extract powder, 15g/L fine jade
Cosmetics, deionized water dissolving, 121 DEG C, moist heat sterilization 30min.
(3) LB liquid medium is prepared: 10g/L tryptone, 10g/L sodium chloride, 5g/L beef extract powder, deionized water
Dissolution, 121 DEG C, moist heat sterilization 30min.
(4) seed liquor that will be prepared, 1/1000 is inoculated in 100 milliliters of liquid LB culture mediums, 160rpm, cultivates 48-
72h, the TB1501 fermentation liquid as prepared.
Embodiment 3
TB1501 prevents and treats powdery mildew of cucumber test:
General × 100TB1501 fermentation liquid dilution, Tebuconazole (being provided by Tianjin Hanbang Plant Protection Product Co., Ltd.),
Flutriafol (being provided by Tianjin Hanbang Plant Protection Product Co., Ltd.) while the cucumber for spraying Powdery Mildew early stage
Blade, setting clear water compare, and spray latter all investigation prevention effects.Efficiency test the results show that TB1501 bacterial strain in anti-Yellow River harnessing
Good control efficiency is played during melon Powdery Mildew, preventive effect is up to 85%, better than chemical agent Tebuconazole (by Tianjin
Han Bang plant protection product Co., Ltd of city provides), almost with pharmaceutical preparations Flutriafol (by Tianjin Chinese nation plant protection product
Co., Ltd provides) quite.
Embodiment 4
Open country powdery mildew of cucumber controlling experiment:
General × 100TB1501 fermentation liquid dilution, Tebuconazole (being provided by Tianjin Hanbang Plant Protection Product Co., Ltd.),
Flutriafol (being provided by Tianjin Hanbang Plant Protection Product Co., Ltd.) while the open country for spraying Powdery Mildew early stage
Cucumber leaves, spray latter all investigation prevention effects.Efficiency test the results show that TB1501 bacterial strain preventive effect up to 80.65%, chemistry
Medicament Flutriafol preventive effect is up to 80.62%, and chemical agent Tebuconazole preventive effect is up to 72.34%.
Embodiment 5
The test of cucumber in greenhouse controlling powdery mildew:
General × 100TB1501 fermentation liquid dilution, Tebuconazole (being provided by Tianjin Hanbang Plant Protection Product Co., Ltd.),
Flutriafol (being provided by Tianjin Hanbang Plant Protection Product Co., Ltd.) while the facility for spraying Powdery Mildew early stage
Cucumber leaves, spray latter all investigation prevention effects.Efficiency test the results show that TB1501 bacterial strain preventive effect up to 85.67%, chemistry
Medicament Flutriafol preventive effect is up to 86.34%, and chemical agent Tebuconazole preventive effect is up to 77.34%.
Claims (6)
1. one plant of Bei Laisi bacillus strain (Bacillus velezensis) Strain Designation be TB1501, it is characterised in that should
Bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation registration number are as follows: CGMCC No.
16069；The preservation time are as follows: on July 6th, 2018.
2. the microbial bacterial agent containing the metabolite for requiring 1 bacterial strain and/or the bacterial strain.
3. microbial bacterial agent as claimed in claim 2, it is characterised in that the microbial bacterial agent is liquid bacterial agent, and in the microbial inoculum
Quantity >=50 of middle viable bacteria and/or living spores~6,000,000,000/g.
4. the preparation method of microbial bacterial agent as claimed in claim 3, it is characterised in that carried out by following step:
(1) activate TB1501 bacterial strain: one ring TB1501 bacterium colony of picking is seeded in 5mL LB liquid medium, 37 DEG C, 160rpm
Shake culture overnight, is made TB1501 seed liquor；
(2) solid LB media is prepared: 10g/L tryptone, 10g/L sodium chloride, 5g/L beef extract powder, 15-20g/L fine jade
Cosmetics, deionized water dissolving, 121 DEG C, moist heat sterilization 30min；
(3) LB liquid medium is prepared: 10g/L tryptone, 10g/L sodium chloride, 5g/L beef extract powder, deionization are water-soluble
Solution, 121 DEG C, moist heat sterilization 30min；
(4) seed liquor that will be prepared, 1/1000 is inoculated in 100 milliliters of liquid LB culture mediums, 160rpm, cultivates 48-72h,
The TB1501 fermentation liquid as prepared.
5. one plant of Bei Laisi bacillus described in claim 1 strain (Bacillus velezensis) Strain Designation is
TB1501, preservation registration number are as follows: application of the CGMCC No. 16069 in prevention and treatment cucumber powdery mildew's pathogen.
6. application of the microbial bacterial agent described in claim 2 in prevention and treatment cucumber powdery mildew's pathogen.
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| CN201811068345.5A CN109022332A (en) | 2018-09-13 | 2018-09-13 | One plant of biological and ecological methods to prevent plant disease, pests, and erosion Bei Laisi bacillus and its application in prevention and treatment cucumber powdery mildew's pathogen |
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Cited By (9)
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| CN110295129A (en) * | 2019-07-23 | 2019-10-01 | 山东省农药科学研究院 | A kind of biocontrol microorganisms and application for preventing and treating gray mold of cucumber and powdery mildew |
| CN110628851A (en) * | 2019-10-16 | 2019-12-31 | 天津市植物保护研究所 | Method for preparing secondary metabolite surfactant of biocontrol bacillus |
| CN110699275A (en) * | 2019-09-03 | 2020-01-17 | 山西省农业科学院农产品贮藏保鲜研究所 | Bacterial strain for antagonizing botrytis cinerea pathogenic bacteria after fruit and vegetable picking and application thereof |
| CN111763629A (en) * | 2019-03-27 | 2020-10-13 | 北京奥沃伽科技有限公司 | Bacillus belgii and application thereof |
| CN113755358A (en) * | 2020-11-30 | 2021-12-07 | 山西农业大学 | Bacillus belgii ZJ1 strain and preparation and use method of fermentation liquor |
| CN114107112A (en) * | 2021-11-29 | 2022-03-01 | 辽宁省蚕业科学研究所 | Biocontrol strain for preventing and treating tussah empty carcass disease and application method thereof |
| CN114410692A (en) * | 2022-01-04 | 2022-04-29 | 海南大学 | Preparation method of n-butanol extract of biocontrol strain HN-2 and application of n-butanol extract in preventing and treating powdery mildew of rubber |
| CN114525227A (en) * | 2022-03-15 | 2022-05-24 | 武汉楚强生物科技有限公司 | Microbial agent containing Bacillus belgii CY30 and application thereof |
| CN114958648A (en) * | 2022-04-06 | 2022-08-30 | 陕西嘉益蓝德生物工程有限公司 | Microbial agent and preparation method thereof |
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Cited By (14)
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| CN111763629B (en) * | 2019-03-27 | 2022-09-20 | 北京奥沃伽科技有限公司 | Bacillus belgii and application thereof |
| CN111763629A (en) * | 2019-03-27 | 2020-10-13 | 北京奥沃伽科技有限公司 | Bacillus belgii and application thereof |
| CN110295129A (en) * | 2019-07-23 | 2019-10-01 | 山东省农药科学研究院 | A kind of biocontrol microorganisms and application for preventing and treating gray mold of cucumber and powdery mildew |
| CN110699275A (en) * | 2019-09-03 | 2020-01-17 | 山西省农业科学院农产品贮藏保鲜研究所 | Bacterial strain for antagonizing botrytis cinerea pathogenic bacteria after fruit and vegetable picking and application thereof |
| CN110699275B (en) * | 2019-09-03 | 2022-11-01 | 山西省农业科学院农产品贮藏保鲜研究所 | Bacterial strain for antagonizing botrytis cinerea pathogenic bacteria after fruit and vegetable picking and application thereof |
| CN110628851A (en) * | 2019-10-16 | 2019-12-31 | 天津市植物保护研究所 | Method for preparing secondary metabolite surfactant of biocontrol bacillus |
| CN113755358A (en) * | 2020-11-30 | 2021-12-07 | 山西农业大学 | Bacillus belgii ZJ1 strain and preparation and use method of fermentation liquor |
| CN113755358B (en) * | 2020-11-30 | 2023-07-04 | 山西农业大学 | Bacillus bailii ZJ1 strain and fermentation broth preparation and use methods |
| CN114107112A (en) * | 2021-11-29 | 2022-03-01 | 辽宁省蚕业科学研究所 | Biocontrol strain for preventing and treating tussah empty carcass disease and application method thereof |
| CN114410692A (en) * | 2022-01-04 | 2022-04-29 | 海南大学 | Preparation method of n-butanol extract of biocontrol strain HN-2 and application of n-butanol extract in preventing and treating powdery mildew of rubber |
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