CN107338211A - One bacillus amyloliquefaciens and its application in forest leaf spot is prevented and treated - Google Patents
One bacillus amyloliquefaciens and its application in forest leaf spot is prevented and treated Download PDFInfo
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- CN107338211A CN107338211A CN201710805526.0A CN201710805526A CN107338211A CN 107338211 A CN107338211 A CN 107338211A CN 201710805526 A CN201710805526 A CN 201710805526A CN 107338211 A CN107338211 A CN 107338211A
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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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Abstract
Application the invention discloses a bacillus amyloliquefaciens and its in forest leaf spot is prevented and treated.The bacterial strain is JRNW0001 1, has been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and deposit number is CGMCC No.13749, and preservation date is on March 10th, 2017.Leaf spot significant effect of the bacillus amyloliquefaciens of the present invention to preventing and treating forest especially ginkgo and beech tree, chemical pesticide is reduced to use, the microbial bacterial agent that bacterial strain JRNW0001 1 is produced can significantly prevent and treat the leaf spot of ginkgo and beech tree, and condition of culture is simple, easily preserve, it is easy to industrialized production, there is good development prospect.
Description
Technical field
The invention belongs to microbial technology field, more particularly to a bacillus amyloliquefaciens and its in preventing and treating forest tikka
Application in disease.
Background technology
Plant disease is one of principal element for influenceing production of forestry, however, the preventing and treating to plant disease is mainly at present
Using chemical agent, after chemical agent long-term use, can to earth zone to remain, pollute, harmful organism develops immunity to drugs,
Injure non-target organism and even destroy the problem of ecological balance etc. is a series of serious, promote people to seek a kind of to the mankind and environment
New plant pest management strategy that is harmless and having good prevention effect.As the public pacifies to chemical pesticide in food and environment
The common concern of full property, makes full use of beneficial microbe controlling plant diseases to be taken seriously increasingly.Especially into 21 century
Since, national governments and the common people more pay attention to the development and application of biological pesticide.Also the development of microbial pesticide and open in China
Hair is classified as state key tackling of key scientific and technical problems and hi-tech development plan.Biological pesticide includes botanical pesticide, and microbial pesticide is micro-
Biological secondary metabolites (antibiotic) etc., for preventing and treating various insects and disease, because it is small, safe to use etc. excellent with endangering
Point, it has also become one of most concerned problem in agricultural sustainable development, it has also become study hotspot.Plant disease biological and ecological methods to prevent plant disease, pests, and erosion for many years is micro-
Biology has been directed to the populations such as fungi, actinomyces, bacterium or even virus (bacteriophage).At present, it is main using more biocontrol bacteria
There are bacillus, Pseudomonas alba, soil radiation bacillus etc..Bacillus amyloliquefaciens (Bacillus
Amyloliquefaciens be) a kind of mesophilous aerobic sporiferous Gram-positive bacillus, can produce it is heat-resisting,
The endospore of drought-enduring, uvioresistant and organic solvent, the scope of its suppression phytopathogen is very wide, including root disease, branch
Fruit disease after dry disease, leaf, flower portion disease and harvest, is a kind of preferable Biocontrol microorganism.
Beech tree, ginkgo are intolerant tree species, the environment of suitable warm slightly humidity, and the two has higher medicinal valency
Value.Bark, the leaf of beech tree can be used as medicine, and have the effect of clearing heat and preventing abortion, and cure mainly flu, headache, stomach real heat, dysentery
Disease, hypogastric pain in pregnancy, anasarca, children's bloody flux, acute conjunctivitis.Ginkgo leaf, food containing gingko almond can be used for palpitation and severe palpitation, deficiency syndrome of the lung cough
The illnesss such as asthma, have beneficial lung qi, control cough and asthma, stop-band worm, reduce just, the dietary function such as flat chapped wrinkle, shield blood vessel, increase CBF and
Medical effect.Due to the medicinal characteristic of beech tree and ginkgo, the chemical agriculture of the unusable high poison of preventing and treating to its pest and disease damage is determined
Medicine.Therefore, it is the task of top priority to find a kind of economic, safe and effective prophylactico-therapeutic measures, and biological control be can yet be regarded as important nondominant hand
Section, wherein biocontrol bacteria plays very important effect in biocontrol of plant disease, and its main advantage is：(1) species
With it is large number of, reproduction speed is fast；Most bacterium colonazition inside and outside plant is strong, largely exists inside and outside plant；(2) it is right
The mode of action variation of pathogen, including competition, antagonism, parasitism, inducing plant resistance generation etc.；(3) can manually cultivate,
It is easily operated, it is easy to production application；(4) some bacterium bacterial strains controlling disease and can not only can promote the growth of plant,
Increase the yield of crop.
Preventing and treating to beech tree, ginkgo leaf spot at present there is no preferable biological and ecological methods to prevent plant disease, pests, and erosion medicament, if biological pesticide or most can be used
Measure few use agricultural chemicals effectively to prevent and treat beech tree, ginkgo leaf spot, residues of pesticides will be reduced to a certain extent, carried significantly
Its high pharmaceutical safety.From the angle preserved the ecological environment with food security, forest production needs low toxicity, pollution less,
Biological agent safe to the human body reaches the purpose being used as medicine safely to control disease, and this also complies with the sustainable development war of forestry
Slightly.
The content of the invention
Goal of the invention：There is the solution starch of antagonism the invention provides one plant to beech tree, ginkgo Leaf blotch pathogeny
Bacillus JRNW0001-1.It is a further object of the present invention to provide the microbial bacterial agent of bacterial strain production and its application, the bacterium
Microbial bacterial agent prepared by strain is remarkably improved beech tree, the ability of ginkgo disease-resistanting leaf spot.
Technical scheme：To achieve the above object, a bacillus amyloliquefaciens (Bacillus as described herein
Amyloliquefaciens), the entitled JRNW0001-1 of bacterial strain, it is general to be preserved in China Committee for Culture Collection of Microorganisms
Logical microorganism center, deposit number is CGMCC No.13749, and preservation date is on March 10th, 2017.Address：Beijing is exposed to the sun
No. 3 Institute of Microorganism, Academia Sinica of institute of area North Star West Road 1, postcode 100101.The bacterial strain is in October, 2014 in Jiangsu
Separation obtains on the disease plant in each forest farm in Jurong City of province, and there is the bacterial strain significant antagonism to make to beech tree, ginkgo leaf spot
With.Bacterial strain JRNW0001-1 Main Biologicals are：For gram-positive bacteria, white, translucent, thalline is shaft-like, bacterium
Fall a diameter of 3-5mm, it is circular, neat in edge, smooth moistening, slightly swell.
Applications of the bacterial strain JRNW0001-1 of the present invention in forest leaf spot is prevented and treated.
Applications of the bacterial strain JRNW0001-1 of the present invention in preventing and treating beech tree or ginkgo leaf spot.
The microbial bacterial agent of bacterial strain JRNW0001-1 productions of the present invention.
The preparation method of the microbial bacterial agent of bacterial strain JRNW0001-1 productions of the present invention, comprises the following steps：
(1) bacillus amyloliquefaciens (Bacillus amyloliquefaciens) JRNW0001-1 is inoculated in fermentation training
Support in base；
(2) the fermented and cultured 40-48 hours at 28-30 DEG C, obtained zymotic fluid is as preventing and treating beech tree and ginkgo leaf spot
Microbial bacterial agent.
Wherein, in step (1), the fermentation medium includes following parts by weight of component：900-1000 parts distilled water, 3-5
Part beef extract, 8-10 parts peptone and 4-5 part sodium chloride.The Medium's PH Value is 6.0-7.0.
The microbial bacterial agent of bacterial strain JRNW0001-1 productions of the present invention is in preventing and treating beech tree and ginkgo leaf spot
Using.
Wherein specific applying step is：By bacillus amyloliquefaciens (Bacillus amyloliquefaciens)
JRNW0001-1 is inoculated in fermentation medium, and fermented and cultured 40 hours at 28-30 DEG C, obtained zymotic fluid is as microorganism
Microbial inoculum, uniformly it is sprayed on ginkgo and beech tree table face.Microbial bacterial agent stoste concentration is 2m L/ strains, is used after 10 times of liquid dilutions
In preventing and treating ginkgo and schneider zelkova leaf pinta, prevention effect is good, and average preventive effect is up to 82.18%.
Beneficial effect：Compared with prior art, it is an advantage of the invention that：
(1) bacillus amyloliquefaciens (Bacillus amyloliquefaciens) JRNW0001-1 is to a variety of trees cause of diseases
Bacterium is inhibited, and very important effect is played in biocontrol bacteria is to forest-crop disease biological control, reduces chemical pesticide
Use, be easy to production application, the especially preventing and treating to fungus-caused beech tree, ginkgo leaf spot significant, ensure
Pharmaceutical safety.
(2) bacillus amyloliquefaciens (Bacillus amyloliquefaciens) JRNW0001-1 condition of culture it is simple,
Easily preserve, be easy to industrialized production, there is good development prospect.
Brief description of the drawings
Fig. 1 is bacterial strain JRNW0001-1 of the present invention to schneider zelkova leaf pinta pathogen alternaria brassica (Alternaria
Brassicae indoor flat plate face-off effect).
Fig. 2 is bacterial strain JRNW0001-1 of the present invention to ginkgo Leaf blotch pathogeny grape spore cup fungi (Botryotinia
Fuckeliana indoor flat plate face-off effect).
Fig. 3 is prevention effect figures of the bacterial strain JRNW0001-1 of the present invention to ginkgo leaf spot.
Fig. 4 is prevention effect figures of the bacterial strain JRNW0001-1 of the present invention to schneider zelkova leaf pinta.
Embodiment
Embodiment 1
Separation obtains a bacillus amyloliquefaciens strain JRNW0001- on the disease plant in each forest farm in Jurong City of Jiangsu Province
1, the bacterial strain is cultivated on the PAD flat boards, and bacterial strain JRNW0001-1 Main Biologicals are：For gram-positive bacteria,
White, translucent, thalline is shaft-like, colony diameter 3-5mm, circular, neat in edge, smooth moistening, slightly swells.The bacterial strain is sent
Friendship is located at BeiJing, China, China Committee for Culture Collection of Microorganisms's common micro-organisms of Institute of Microorganism, Academia Sinica
Center (abbreviation CGMCC) preservation, deposit number are CGMCC No.13749, and preservation date is on March 10th, 2017.It is identified,
The bacterial strain is bacillus amyloliquefaciens, is named as JRNW0001-1.
Embodiment 2
The microbial bacterial agent of bacillus amyloliquefaciens JRNW0001-1 productions：By 4g beef extracts, 8 parts of peptones and 4.5g parts
Sodium chloride is dissolved in 1000ml distilled water, and regulation pH is 6.0, is stirred, and sterilizes, obtains fermentation medium, will solve starch gemma bar
Bacterium (Bacillus amyloliquefaciens) JRNW0001-1 is inoculated in fermentation medium, training of being fermented at 28-30 DEG C
Support 40-48 hours, obtained zymotic fluid, as microbial bacterial agent.
Embodiment 3
The microbial bacterial agent of bacillus amyloliquefaciens (Bacillus amyloliquefaciens) JRNW0001-1 productions
In the effect analysis of preventing and treating beech tree and ginkgo leaf spot.
(1) laboratory test
By with the bacillus amyloliquefaciens of LB slant activations (Bacillus amyloliquefaciens) JRNW0001-1
Bacterial strain is symmetrically inoculated in the both sides of PDA plate, then with diameter 5mm card punch respectively by schneider zelkova leaf pinta pathogen rape chain lattice
The pathogen grape spore cup fungi (Botryotinia fuckeliana) of spore (Alternaria brassicae), ginkgo leaf spot
PDA plate culture break into bacteria cake, be inoculated in flat board center, 28 ± 2 DEG C culture 10 days observation fungistatic effect, measure solution starch
Bacillus (Bacillus amyloliquefaciens) JRNW0001-1 is to schneider zelkova leaf pinta pathogen alternaria brassica
(Alternaria brassicae), ginkgo Leaf blotch pathogeny grape spore cup fungi (Botryotinia fuckeliana) are put down
Plate bacteriostasis, control is used as by the use of the PDA plate for singly connecing pathogen.
Experiment effect as shown in Figure 1, Figure 2, shown in table 1.A in Fig. 1 is bacterial strain JRNW0001-1 to schneider zelkova leaf pinta pathogen
The indoor flat plate face-off result of alternaria brassica (Alternaria brassicae), the B in Fig. 1 is schneider zelkova leaf pinta pathogen
Alternaria brassica (Alternaria brassicae) single inoculation figure, it can be seen that bacterial strain JRNW0001-1 is to alternaria brassica
Growth has significant inhibition.
A in Fig. 2 is bacterial strain JRNW0001-1 to ginkgo Leaf blotch pathogeny grape spore cup fungi (Botryotinia
Fuckeliana indoor flat plate face-off result), the B in Fig. 2 is ginkgo Leaf blotch pathogeny grape spore cup fungi
(Botryotinia fuckeliana) single inoculation figure, it can be seen that bacterial strain JRNW0001-1 of the present invention is to grape spore cup fungi
Growth has significant inhibition.
As can be seen from the above results, bacillus amyloliquefaciens (Bacillus amyloliquefaciens)
JRNW0001-1 has obvious antagonism to 2 kinds of pathogens.
Inhibitions of the JRNW0001-1 of table 1 to pathogen
(2) application effect determines
Germ spore suspension is made in schneider zelkova leaf pinta pathogen alternaria brassica (Alternaria brassicae),
Spore concentration is 2 × 105Individual/ml；Ginkgo Leaf blotch pathogeny grape spore cup fungi (Botryotinia fuckeliana) is made
Germ spore suspension, spore concentration are 2 × 105Individual/ml.
Artificial infection experiment is carried out in greenhouse.At the top of the beech tree tree crown that growth period is 1 year, manually manufactured with needle point method
Wound, in the spore suspension 2ml/ strains of the surface sprinkling pathogen alternaria brassica of manufacture wound；Then sprinkling solution starch bud
Spore bacillus JRNW0001-1 microbial bacterial agents, microbial bacterial agent dosage are 2ml/ strains, are used after 10 times of liquid dilutions.To be only inoculated with disease
Opportunistic pathogen, be not inoculated with bacillus amyloliquefaciens JRNW0001-1 microbial bacterial agents is used as control (CK), every kind of processing for examination plant
It is 100 plants.After artificial infection in 5 days, relative humidity 60% is kept in greenhouse, in favor of pathogen and the micro- lifes of JRNW0001-1
The survival of thing microbial inoculum and smoothly intrusion are inoculated with plant.
Artificial infection processing is carried out to ginkgo plant using with above identical method.
The occurring degree of leaf spot, the statistics incidence of disease, disease index are monitored after inoculation daily.
Diseased plant grade scale：Mainly there is Lesion number and diameter according to inoculation blade tow sides in severity Scaling, just fixed
Grade scale is：
0：Normally；
Ⅰ：Tow sides have a small amount of scab；
Ⅱ：Tow sides have scab, and onset area accounts for the blade gross area below 1/4；
Ⅲ：Tow sides have scab, and onset area accounts for the 1/4-1/2 of the blade gross area；
Ⅳ：Tow sides have scab, and onset area accounts for blade gross area 1/2-3/4；
Ⅴ：Tow sides have scab, and onset area accounts for the blade gross area more than 3/4.
(total strain number × highest level represents number to disease-sensitive index=∑ (the diseased plant numbers at different levels × level represents numerical value) × 100/
Value)；
Susceptible gene=(susceptible strain tree/inoculation strain tree) × 100%.
It is Fig. 3, shown respectively in the 5th, 10,15,20,25,30 day statistics susceptible gene and disease-sensitive index after artificial infection
Bacterial strain JRNW0001-1 microbial bacterial agents of the present invention are to the prevention effect of ginkgo leaf spot, the bacterial strain of the present invention shown in Fig. 4
Prevention effect of the JRNW0001-1 microbial bacterial agents to schneider zelkova leaf pinta, it can be seen that with the time after artificial infection pathogen
Extension, be inoculated with the ginkgoes of JRNW0001-1 microbial bacterial agents, beech tree and do not connect at the control of RNW0001-1 microbial bacterial agents
In reason, the susceptible gene and disease-sensitive index of plant leaf spot all progressively increase, but the processing for connecing RNW0001-1 microbial bacterial agents is notable
Less than the control treatment for not connecing JRNW0001-1 microbial bacterial agents, show to be inoculated with RNW0001-1 microbial bacterial agents to ginkgo, beech tree
Leaf spot have preferable preventive effect.
Claims (8)
1. a bacillus amyloliquefaciens (Bacillus amyloliquefaciens), the entitled JRNW0001-1 of bacterial strain, have been protected
It is hidden in China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number is CGMCC No.13749, preservation
Date is on March 10th, 2017.
2. applications of the bacterial strain JRNW0001-1 in forest leaf spot is prevented and treated described in claim 1.
3. application according to claim 2, it is characterised in that the forest is beech tree or ginkgo.
A kind of 4. microbial bacterial agent that bacterial strain JRNW0001-1 using described in claim 1 is produced.
5. the preparation method of the microbial bacterial agent described in a kind of claim 4, it is characterised in that comprise the following steps：
(1) bacillus amyloliquefaciens (Bacillus amyloliquefaciens) JRNW0001-1 is inoculated in fermentation medium
In；
(2) the fermented and cultured 40-48 hours at 28-30 DEG C, obtained zymotic fluid as preventing and treating beech tree and ginkgo leaf spot it is micro-
Bacteria agent.
6. preparation method according to claim 5, it is characterised in that in step (1), the fermentation medium includes following
Parts by weight of component：900-1000 parts distilled water, 3-5 parts beef extract, 8-10 parts peptone and 4-5 part sodium chloride.
7. preparation method according to claim 6, it is characterised in that the Medium's PH Value is 6.0-7.0.
8. application of the microbial bacterial agent as claimed in claim 4 in preventing and treating beech tree or ginkgo leaf spot.
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| CN201710805526.0A CN107338211A (en) | 2017-09-08 | 2017-09-08 | One bacillus amyloliquefaciens and its application in forest leaf spot is prevented and treated |
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| CN201710805526.0A CN107338211A (en) | 2017-09-08 | 2017-09-08 | One bacillus amyloliquefaciens and its application in forest leaf spot is prevented and treated |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113201478A (en) * | 2021-05-27 | 2021-08-03 | 广西民族大学 | Siamese bacillus Gxun-36 secreting iron carrier and application thereof |
| CN115125159A (en) * | 2022-04-24 | 2022-09-30 | 沈阳农业大学 | Bacillus amyloliquefaciens and microbial inoculum thereof, and preparation method and application of microbial inoculum |
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| CN103695356A (en) * | 2013-12-24 | 2014-04-02 | 河北省科学院生物研究所 | Bacillus amyloliquefaciens, microbial inoculant and application thereof |
| CN105219681A (en) * | 2015-11-04 | 2016-01-06 | 湖北工程学院 | A kind of bacillus amyloliquefaciens Bacillus amyloliquefaciens D2WM and preparation method and application |
| CN105316264A (en) * | 2015-12-04 | 2016-02-10 | 北京农学院 | Bacillus amyloliquefaciens and application thereof |
| CN105316265A (en) * | 2015-12-04 | 2016-02-10 | 北京农学院 | Bacillus amyloliquefaciens used for biocontrol and application thereof |
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2017
- 2017-09-08 CN CN201710805526.0A patent/CN107338211A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103695356A (en) * | 2013-12-24 | 2014-04-02 | 河北省科学院生物研究所 | Bacillus amyloliquefaciens, microbial inoculant and application thereof |
| CN105219681A (en) * | 2015-11-04 | 2016-01-06 | 湖北工程学院 | A kind of bacillus amyloliquefaciens Bacillus amyloliquefaciens D2WM and preparation method and application |
| CN105316264A (en) * | 2015-12-04 | 2016-02-10 | 北京农学院 | Bacillus amyloliquefaciens and application thereof |
| CN105316265A (en) * | 2015-12-04 | 2016-02-10 | 北京农学院 | Bacillus amyloliquefaciens used for biocontrol and application thereof |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113201478A (en) * | 2021-05-27 | 2021-08-03 | 广西民族大学 | Siamese bacillus Gxun-36 secreting iron carrier and application thereof |
| CN115125159A (en) * | 2022-04-24 | 2022-09-30 | 沈阳农业大学 | Bacillus amyloliquefaciens and microbial inoculum thereof, and preparation method and application of microbial inoculum |
| CN115125159B (en) * | 2022-04-24 | 2023-05-23 | 沈阳农业大学 | Bacillus amyloliquefaciens, microbial inoculum thereof, preparation method and application of microbial inoculum |
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Application publication date: 20171110 |
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