CN106282067A - Multifunctional agricultural complex micro organism fungicide and microbial ecological agent and application - Google Patents
Multifunctional agricultural complex micro organism fungicide and microbial ecological agent and application Download PDFInfo
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- CN106282067A CN106282067A CN201610718598.7A CN201610718598A CN106282067A CN 106282067 A CN106282067 A CN 106282067A CN 201610718598 A CN201610718598 A CN 201610718598A CN 106282067 A CN106282067 A CN 106282067A
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- 239000003795 chemical substances by application Substances 0.000 title claims abstract description 103
- 230000000813 microbial effect Effects 0.000 title claims abstract description 100
- 244000005700 microbiome Species 0.000 title claims abstract description 51
- 239000000417 fungicide Substances 0.000 title claims abstract description 38
- 230000000855 fungicidal effect Effects 0.000 title claims abstract description 37
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims abstract description 104
- 241000894006 Bacteria Species 0.000 claims abstract description 45
- 241000193755 Bacillus cereus Species 0.000 claims abstract description 39
- 241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 35
- 241000194108 Bacillus licheniformis Species 0.000 claims abstract description 34
- 241000193738 Bacillus anthracis Species 0.000 claims abstract description 29
- 241000196324 Embryophyta Species 0.000 claims abstract description 25
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- 238000002360 preparation method Methods 0.000 claims description 16
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- 230000002265 prevention Effects 0.000 claims description 7
- 230000001580 bacterial effect Effects 0.000 claims description 6
- 239000002068 microbial inoculum Substances 0.000 claims description 5
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- 239000002131 composite material Substances 0.000 claims description 4
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- 239000012530 fluid Substances 0.000 description 4
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- 239000000463 material Substances 0.000 description 4
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- 239000001888 Peptone Substances 0.000 description 2
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- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
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- VTYYLEPIZMXCLO-UHFFFAOYSA-L calcium carbonate Substances [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
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- LGZXYFMMLRYXLK-UHFFFAOYSA-N mercury(2+);sulfide Chemical compound [S-2].[Hg+2] LGZXYFMMLRYXLK-UHFFFAOYSA-N 0.000 description 2
- 239000003960 organic solvent Substances 0.000 description 2
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- 241001310537 Acetobacter orientalis Species 0.000 description 1
- 239000005995 Aluminium silicate Substances 0.000 description 1
- 241000976738 Bacillus aryabhattai Species 0.000 description 1
- 241000278457 Bacillus siamensis Species 0.000 description 1
- 240000007124 Brassica oleracea Species 0.000 description 1
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- 235000005755 Vigna unguiculata ssp. sesquipedalis Nutrition 0.000 description 1
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- 230000007850 degeneration Effects 0.000 description 1
- GUJOJGAPFQRJSV-UHFFFAOYSA-N dialuminum;dioxosilane;oxygen(2-);hydrate Chemical compound O.[O-2].[O-2].[O-2].[Al+3].[Al+3].O=[Si]=O.O=[Si]=O.O=[Si]=O.O=[Si]=O GUJOJGAPFQRJSV-UHFFFAOYSA-N 0.000 description 1
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- NLYAJNPCOHFWQQ-UHFFFAOYSA-N kaolin Chemical compound O.O.O=[Al]O[Si](=O)O[Si](=O)O[Al]=O NLYAJNPCOHFWQQ-UHFFFAOYSA-N 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 235000019713 millet Nutrition 0.000 description 1
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- 235000010446 mineral oil Nutrition 0.000 description 1
- 230000002073 mitogenetic effect Effects 0.000 description 1
- 229910052901 montmorillonite Inorganic materials 0.000 description 1
- DIOQZVSQGTUSAI-UHFFFAOYSA-N n-butylhexane Natural products CCCCCCCCCC DIOQZVSQGTUSAI-UHFFFAOYSA-N 0.000 description 1
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- 239000003002 pH adjusting agent Substances 0.000 description 1
- 239000010695 polyglycol Substances 0.000 description 1
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- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
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- 235000012222 talc Nutrition 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 239000000052 vinegar Substances 0.000 description 1
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Classifications
-
- 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
-
- 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
-
- 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
- A01N63/10—Animals; Substances produced thereby or obtained therefrom
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- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Virology (AREA)
- Chemical & Material Sciences (AREA)
- Genetics & Genomics (AREA)
- Pest Control & Pesticides (AREA)
- Dentistry (AREA)
- Agronomy & Crop Science (AREA)
- Environmental Sciences (AREA)
- Organic Chemistry (AREA)
- Plant Pathology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses multifunctional agricultural complex micro organism fungicide and microbial ecological agent and application.Agricultural complex micro organism fungicide provided by the present invention, its active component is made up of A Yebo plurispore bacillus, Bacillus licheniformis, Lyceum bacillus cereus and east acetic acid bacteria.Agricultural microbial ecological agent provided by the present invention, its active component is to cultivate, with fermentation medium, the culture that the agricultural complex micro organism fungicide of preceding claim obtains.Above-mentioned agricultural complex micro organism fungicide and above-mentioned agricultural microbial ecological agent can be used for preventing and/or treat plant anthrax, can be used for promoting crop yield.
Description
Technical field
The present invention relates in agricultural and microorganism field multifunctional agricultural complex micro organism fungicide and microbial ecological agent with should
With.
Background technology
China is vast in territory, and weather conditions are complicated, and the fruit tree of plantation is of a great variety, the fruit tree that cultivated area is bigger have Fructus Persicae,
Fructus Mali pumilae etc..Fructus Persicae and apple disease kind are the most more.Peach tree anthrax, is the important fruit disease of peach tree.The master of peach tree anthrax
Endangering is fruit, it is possible to infect children's tip and blade.Young fruit is caught an illness to grow and is stopped, and really face crineous, it is residual that atrophy hardens into mummy
Stay on branch.After Fruit, attacked fruit face is just in filbert water stain shape scab, and the rear brown that reddens that expands, scab depression has bright
Aobvious concentric wheel stripe shape wrinkle, produces Chinese red clayey granule point during high humidity, the soft corruption of last sick fruit comes off or forms mummy residual
On branch.Young sprout is caught an illness, and cave in scab in oblong brown, and sick tip lateral thrust is withered time serious.Blade catch an illness generation light
Circular or the irregular shape taupe scab of brown, it produces Chinese red to black grain point.Withered the coming off of rear scab wears gift, young sprout
Top vane atrophy is sagging, indulges and is rolled into tubulose.
Along with the public is more and more higher to the requirement of " pollution-free food ", " pollution-free food ", market to pollution-free, can naturally drop
The ecological agricultural chemical demand solved the most gradually increases.From the point of view of the development of pesticide science, on pesticide manufacturing uses " back to nature "
And " Harmless " of pesticide, it is the inexorable trend of society and Development of Natural Science, therefore new bio wide-spectrum bactericide has
Wide development prospect.
Summary of the invention
The technical problem to be solved is how to prevent and/or treat plant anthrax and/or promote that crop increases
Produce.
In order to solve above technical problem, the invention provides agricultural complex micro organism fungicide.
Agricultural complex micro organism fungicide provided by the present invention, its active component is by A Yebo plurispore bacillus, lichens
Bacillus cereus, Lyceum bacillus cereus and east acetic acid bacteria composition.
In above-mentioned agricultural complex micro organism fungicide, described A Yebo plurispore bacillus is A Yebo plurispore bacillus ACCC
19947, described Bacillus licheniformis is Bacillus licheniformis ACCC 19946, and described Lyceum bacillus cereus is Lyceum bacillus cereus
ACCC 19948, described east acetic acid bacteria is east acetic acid bacteria ACCC 19949.
In above-mentioned agricultural complex micro organism fungicide, A Yebo plurispore bacillus, Bacillus licheniformis, Lyceum bacillus cereus and
The colony-forming units ratio of east acetic acid bacteria can be (2.5-4): 1:(1-2): (3-4.5).
In above-mentioned agricultural complex micro organism fungicide, A Yebo plurispore bacillus, Bacillus licheniformis, Lyceum bacillus cereus and
The colony-forming units ratio of east acetic acid bacteria can be 3.25:1:1.5:3.75.
Above-mentioned agricultural complex micro organism fungicide can be used for preventing and/or treat plant anthrax, can be used for promoting that crop increases
Produce.
Described A Yebo plurispore bar in above-mentioned agricultural complex micro organism fungicide or described agricultural complex micro organism fungicide
Any one application following of bacterium, Bacillus licheniformis, Lyceum bacillus cereus and east acetic acid bacteria falls within the protection model of the present invention
Enclose:
1) application in preparation plant Anthracnose Pathogen bacteria inhibitor;
2) application in prevention and/or treatment plant anthrax;
3) application in promoting crop yield;
4) application in preparing following agricultural microbial ecological agent.
In order to solve above technical problem, present invention also offers agricultural microbial ecological agent.
Agricultural microbial ecological agent provided by the present invention, its active component is to cultivate described agricultural multiple with fermentation medium
Close the culture (product in culture vessel) that microbial bacterial agent obtains.
In above-mentioned agricultural microbial ecological agent, A Yebo plurispore bacillus, Bacillus licheniformis, Lyceum bacillus cereus and east
The colony-forming units of acetic acid bacteria is than being (10-30): 1:(1-5): (30-50) or 18:1:1.6:40.
In above-mentioned agricultural microbial ecological agent, described fermentation medium can be the molasses water solution of variable concentrations, such as 6% (matter
Amount percentage composition) molasses water solution.
Molasses are the side-products of sugar industry.The general brix of molasses is 80-90Bx, containing sugar more than 50%.
In above-mentioned agricultural microbial ecological agent, described fermentation medium cultivates what described agricultural complex micro organism fungicide used
Cultivation temperature can be 20-30 DEG C.
Above-mentioned agricultural microbial ecological agent can be used for preventing and/or treat plant anthrax, can be used for promoting crop yield.
Any one application following of above-mentioned agricultural microbial ecological agent falls within protection scope of the present invention:
1) application in preparation plant Anthracnose Pathogen bacteria inhibitor;
2) application in prevention and/or treatment plant anthrax;
3) application in promoting crop yield.
Above, described plant can be rosaceous plant, and described crop can be Theaceae crop, solanaceous crops, legume
Or crop in cruciferae.
Described rosaceous plant can be Fructus Persicae, and described Theaceae crop can be tea, and described solanaceous crops can be Fructus Capsici, described pulse family
Crop can be Semen vignae sinensis, and described crop in cruciferae can be Caulis et Folium Brassicae capitatae.
Above-mentioned agricultural complex micro organism fungicide and above-mentioned agricultural microbial ecological agent may also include carrier, and preferably pesticide field is normal
And be inert carrier biologically.Described carrier can be solid carrier or liquid-carrier.Described solid carrier is ore deposit
Thing material, vegetable material or macromolecular compound;Described mineral material is clay, Talcum, Kaolin, montmorillonite, white carbon, boiling
At least one in stone, Silicon stone and kieselguhr;Described vegetable material is at least one in Semen Maydis powder, Semen Glycines powder and starch;Described
Macromolecular compound is polyvinyl alcohol and/or polyglycols.Described liquid-carrier is organic solvent, vegetable oil, mineral oil or water;Institute
Stating organic solvent is decane and/or dodecane.In described microbial inoculum, described active component can with the living cells being cultured, work carefully
Presented in the mixture of the fermentation liquid of born of the same parents, the filtrate of cell culture or cell and filtrate.Described living cells can be mitogenetic
Spore, chlamydospore, mycelia or containing conidium and the mycelial form of mycelia, preferably conidium or chlamydospore
Form.The dosage form of described compositions can be multiple dosage form, such as liquor, Emulsion, suspending agent, powder, granule, wettable powder
Or water dispersible granules.
As required, above-mentioned agricultural complex micro organism fungicide and above-mentioned agricultural microbial ecological agent also can add surface activity
Agent (such as polysorbas20, Tween 80 etc.), binding agent, stabilizer (such as antioxidant), pH adjusting agent etc..
It is demonstrated experimentally that cultivate, with fermentation medium, the agricultural microbial ecological agent pair that described agricultural complex micro organism fungicide obtains
The preventive effect of peach tree anthrax reaches 92.39%, and the therapeutic effect of peach tree anthrax is reached 83.65%, the increasing to Folium Camelliae sinensis
Productivity reaches 12%, the rate of growth difference 20% and 15% to Fructus Capsici, Semen vignae sinensis, makes cabbage head diameter increase 4-6cm.
Detailed description of the invention
Being further described in detail the present invention below in conjunction with detailed description of the invention, the embodiment be given is only for explaining
The bright present invention rather than in order to limit the scope of the present invention.Experimental technique in following embodiment, if no special instructions, is
Conventional method.Material used in following embodiment, reagent etc., if no special instructions, the most commercially obtain.
A Yebo plurispore bacillus ACCC 19947 in following embodiment is A Yebo plurispore bacillus (Bacillus
Aryabhattai) R002, is concealed in China Committee for Culture Collection of Microorganisms's agricultural microorganism on August 1st, 2016
ACCC, address: No.12 ,zhongguancun south street,Haidian District, Beijing, Chinese Academy of Agricultural Sciences's agricultural resource and agricultural (are called for short in center
Research on zoning institute, postcode 100081), from this collection day, the public can be micro-from China Committee for Culture Collection of Microorganisms's agricultural
Bio-Centers obtains this bacterial strain.
Bacillus licheniformis ACCC 19946 in following embodiment is Bacillus licheniformis (Bacillus
Licheniformis) R001, is concealed in China Committee for Culture Collection of Microorganisms's micro-life of agricultural on August 1st, 2016
ACCC, address: No.12 ,zhongguancun south street,Haidian District, Beijing, Chinese Academy of Agricultural Sciences's agricultural resource and agriculture (are called for short in thing center
Industry Research on zoning institute, postcode 100081), from this collection day, the public can be from China Committee for Culture Collection of Microorganisms's agricultural
Microorganism center obtains this bacterial strain.
Lyceum bacillus cereus ACCC 19948 in following embodiment is Lyceum bacillus cereus (Bacillus
Siamensis) R003, is concealed in China Committee for Culture Collection of Microorganisms's agricultural microorganism on August 1st, 2016
The heart (is called for short ACCC, address: No.12 ,zhongguancun south street,Haidian District, Beijing, Chinese Academy of Agricultural Sciences's agricultural resource and farming region
Draw institute, postcode 100081), from this collection day, the public can be from China Committee for Culture Collection of Microorganisms's micro-life of agricultural
Thing center obtains this bacterial strain.
East acetic acid bacteria ACCC 19949 in following embodiment is east acetic acid bacteria (Acetobacter
Orientalis) R004, is concealed in China Committee for Culture Collection of Microorganisms's agricultural microorganism on August 1st, 2016
ACCC, address: No.12 ,zhongguancun south street,Haidian District, Beijing, Chinese Academy of Agricultural Sciences's agricultural resource and agricultural (are called for short in center
Research on zoning institute, postcode 100081), from this collection day, the public can be micro-from China Committee for Culture Collection of Microorganisms's agricultural
Bio-Centers obtains this bacterial strain.
A Yebo plurispore bacillus ACCC 19947, Bacillus licheniformis ACCC 19946 and Lyceum bacillus cereus ACCC
19948 use nutrient culture medium culturing.Nutrient culture medium is as follows:
Nutrient solid medium: peptone 5g, NaCl 5g, meat extract 3g, agar 15g distilled water is settled to
1000ml, pH7.0.Sterilizing 20 minutes under the conditions of 121 DEG C.
Nutrient fluid medium: peptone 5g, NaCl 5g, meat extract 3g, is settled to 1000ml with distilled water,
pH7.0.Sterilizing 20 minutes under the conditions of 121 DEG C.
East acetic acid bacteria ACCC 19949 uses acetic acid bacteria culture medium culturing, and acetic acid bacteria culture medium is as follows:
Acetic acid bacteria solid medium: glucose 100g, yeast extract 10g, CaCO320g, agar 15g, use distilled water constant volume
To 1000ml, pH6.8.Sterilizing 20 minutes under the conditions of 121 DEG C.
Acetic acid bacteria fluid medium is: glucose 100g, yeast extract 10g, CaCO320g, is settled to distilled water
1000ml, pH6.8.Sterilizing 20 minutes under the conditions of 121 DEG C.
Embodiment 1, the preparation of multifunctional agricultural complex micro organism fungicide
1, dormancy actication of culture
By the A Yebo plurispore bacillus ACCC 19947 of resting state, Bacillus licheniformis ACCC 19946, Lyceum bud
Spore bacillus ACCC 19948 and east acetic acid bacteria ACCC 19949 is respectively connected to test tube slant activation;A Yebo plurispore bacillus
ACCC 19947, Bacillus licheniformis ACCC 19946 and Lyceum bacillus cereus ACCC 19948 use nutrient solid culture
Base is cultivated, cultivation temperature 28-30 DEG C, cultivation 24-48 hour;East acetic acid bacteria ACCC 19949 uses acetic acid bacteria solid medium
Cultivate, cultivation temperature 28-30 DEG C, cultivate 24-48 hour continuously.
2, the preparation of multifunctional agricultural complex micro organism fungicide
This step provides two kinds of multifunctional agricultural complex micro organism fungicides, respectively multifunctional agricultural composite microbial bacteria
Agent A and multifunctional agricultural complex micro organism fungicide B.The active component of multifunctional agricultural complex micro organism fungicide A is by A Yeboduo
Bacillus cereus ACCC 19947, Bacillus licheniformis ACCC 19946, Lyceum bacillus cereus ACCC 19948 and east acetic acid bacteria
ACCC 19949 forms.In this multifunctional agricultural complex micro organism fungicide A, A Yebo plurispore bacillus ACCC 19947, lichens
Bacillus cereus ACCC 19946, Lyceum bacillus cereus ACCC 19948 and the colony-forming units of east acetic acid bacteria ACCC 19949
(cfu) ratio is 13:4:6:15.The active component of multifunctional agricultural complex micro organism fungicide B is by A Yebo plurispore bacillus ACCC
19947, Bacillus licheniformis ACCC 19946 and Lyceum bacillus cereus ACCC 19948 forms.This multifunctional agricultural composite microbial
In thing microbial inoculum B, A Yebo plurispore bacillus ACCC 19947, Bacillus licheniformis ACCC 19946 and Lyceum bacillus cereus ACCC
Colony-forming units (cfu) ratio of 19948 is 13:4:6.
Concrete preparation method is as follows:
A Yebo plurispore bacillus ACCC 19947 after step 1 is activated access equipped with sterilizing after nutrient liquid
The shaking flask (equipped with 100mL culture medium, rotating speed is 180rpm to 500ml triangular flask) of culture medium, Bacillus licheniformis ACCC 19946
(500ml triangular flask equipped with 100mL culture medium, rotating speed is to access the shaking flask equipped with the nutrient fluid medium after sterilizing
Shaking flask (the 500ml of the nutrient fluid medium after 180rpm), Lyceum bacillus cereus ACCC 19948 accesses equipped with sterilizing
Triangular flask is equipped with 100mL culture medium, and rotating speed is 180rpm), east acetic acid bacteria ACCC 19949 access equipped with sterilizing after acetic acid
The shaking flask (equipped with 100mL culture medium, rotating speed is 180rpm to 500ml triangular flask) of bacteria liquid culture medium, is then placed in constant temperature oscillation
Device is cultivated, temperature 28-30 DEG C, cultivation 15-48 hour, respectively obtains the liquid of A Yebo plurispore bacillus ACCC 19947
Strain, the liquid spawn of Bacillus licheniformis ACCC 19946, the liquid spawn of Lyceum bacillus cereus ACCC 19948, east vinegar
The liquid spawn of acid bacterium ACCC 19949.A Yebo polygerm in the liquid spawn of this A Yebo plurispore bacillus ACCC 19947
Spore bacillus ACCC 19947 content is 13 × 108Cfu/ml, the ground in the liquid spawn of this Bacillus licheniformis ACCC 19946
Clothing bacillus cereus ACCC 19946 content is 4 × 108Cfu/ml, this Lyceum bacillus cereus ACCC 19948 liquid spawn in
Lyceum bacillus cereus ACCC 19948 content is 6 × 108Cfu/ml, east acetic acid bacteria ACCC 19949 liquid spawn in east
Side's acetic acid bacteria ACCC 19949 content is 15 × 108cfu/ml。
By liquid spawn, the liquid of Bacillus licheniformis ACCC 19946 of above-mentioned A Yebo plurispore bacillus ACCC 19947
The liquid spawn of body strain, the liquid spawn of Lyceum bacillus cereus ACCC 19948 and east acetic acid bacteria ACCC 19949 according to etc.
Volume mixture obtains multifunctional agricultural complex micro organism fungicide A.In this multifunctional agricultural complex micro organism fungicide A, A Yeboduo
Bacillus cereus ACCC 19947, Bacillus licheniformis ACCC 19946, Lyceum bacillus cereus ACCC 19948 and east acetic acid bacteria
Colony-forming units (cfu) ratio of ACCC 19949 is 13:4:6:15.
By liquid spawn, the liquid of Bacillus licheniformis ACCC 19946 of above-mentioned A Yebo plurispore bacillus ACCC 19947
The liquid spawn of body strain and Lyceum bacillus cereus ACCC 19948 is mixed to get multifunctional agricultural composite microbial according to equal-volume
Thing microbial inoculum B.In this multifunctional agricultural complex micro organism fungicide B, A Yebo plurispore bacillus ACCC 19947, Bacillus licheniformis
Colony-forming units (cfu) ratio of ACCC 19946 and Lyceum bacillus cereus ACCC 19948 is 13:4:6.
Embodiment 2, the preparation of multifunctional agricultural microbial ecological agent
Present embodiments provide two kinds of multifunctional agricultural microbial ecological agent, be respectively multifunctional agricultural microbial ecological agent A and
Multifunctional agricultural microbial ecological agent B.
1, the preparation of multifunctional agricultural microbial ecological agent A
In 100 liters of seed tanks, 50 liters of concentration of addition are the molasses water solution of 6% (weight/mass percentage composition), after sterilizing, and will
The multifunctional agricultural complex micro organism fungicide A of embodiment 1 accesses in these 100 liters of seed tanks, and inoculum concentration is 0.8%-1%, ventilation
Amount is 0.5 (V/V min) (ratio that the volume of air being passed through fermentation tank per minute is amassed with fermentation cylinder for fermentation liquid), rotating speed
For 200rpm, cultivate 24 hours under the conditions of 30 DEG C, obtain seed liquor.
In 1000 liters of fermentation tanks, 500 liters of concentration of addition are the molasses water solution of 6% (weight/mass percentage composition), after sterilizing,
Seed liquor is accessed in 1000 liters of fermentation tanks and fermented, and inoculum concentration is that 10% (seed liquor is culture volume in fermentation tank
10%), ventilation ratio ventilation is 0.5 (V/V min) (volume of air being passed through fermentation tank per minute and fermentation cylinder for fermentation liquid
The ratio of volume), rotating speed 200rpm, control pH7.0-7.2,30 DEG C cultivate 24 hours, add volume be liquid in fermentation tank
The molasses of the 4% of volume carry out feed supplement for the first time, and uncomfortable pH after feed supplement for the first time is complete, the constant continuation of temperature, rotating speed, ventilation is sent out
Ferment adds volume after within 18 hours, arriving stable phase be that in fermentation tank, the molasses of the 5% of liquid volume carry out secondary feed supplement, and secondary is mended
Having expected to adjust pH to 4.5-5.0, other parameter constant continues to ferment and reaches stable phase in 18 hours, by the bacterium solution of this stable phase and aseptic
Water is mixed to get multifunctional agricultural microbial ecological agent A, and wherein bacterium solution is 1:4 with the volume ratio of sterilized water.This multifunctional agricultural is micro-
In ecological agent A, A Yebo plurispore bacillus ACCC 19947 content is 9 × 108Cfu/mL, Lyceum bacillus cereus ACCC
19948 content are 8.0 × 107Cfu/mL, Bacillus licheniformis ACCC 19946 content is 5.0 × 107Cfu/mL, east acetic acid
Bacterium ACCC 19949 content is 2 × 109cfu/mL.In multifunctional agricultural microbial ecological agent A, A Yebo plurispore bacillus ACCC
19947, Bacillus licheniformis ACCC 19946, Lyceum bacillus cereus ACCC 19948 and the bacterium of east acetic acid bacteria ACCC 19949
Falling to being formed unit (cfu) ratio is 18:1:1.6:40.
2, the preparation of multifunctional agricultural microbial ecological agent B
Multifunctional agricultural complex micro organism fungicide A by the embodiment 1 in step 1 replaces with the multi-functional agriculture of embodiment 1
Using complex micro organism fungicide B, other is constant, obtains multifunctional agricultural microbial ecological agent B.In multifunctional agricultural microbial ecological agent B,
A Yebo plurispore bacillus ACCC 19947 content is 9 × 108Cfu/mL, Lyceum bacillus cereus ACCC 19948 content is 8.0
×107Cfu/mL, Bacillus licheniformis ACCC 19946 content is 5.0 × 107cfu/mL.Multifunctional agricultural microbial ecological agent B
In, A Yebo plurispore bacillus ACCC 19947, Bacillus licheniformis ACCC 19946 and Lyceum bacillus cereus ACCC 19948
Colony-forming units than for 18:1:1.6.
Embodiment 3, multifunctional agricultural microbial ecological agent are to the prevention of peach tree anthrax and treatment
Use 200 times of diluents and the multifunctional agricultural of embodiment 2 of the multifunctional agricultural microbial ecological agent A of embodiment 2
200 times of diluents of microbial ecological agent B carry out prevention and the treatment of peach tree anthrax.The multifunctional agricultural Tiny ecosystem of embodiment 2
200 times of diluents of preparation A are, with sterilized water, the multifunctional agricultural microbial ecological agent A of embodiment 2 is diluted 200 times of liquid obtained
Body.200 times of diluents of the multifunctional agricultural microbial ecological agent B of embodiment 2 are by the multifunctional agricultural of embodiment 2 with sterilized water
Microbial ecological agent B dilutes 200 times of liquid obtained.
1, the prevention of peach tree anthrax
In a peach garden of the Shandong Province that all can be attacked in various degree by anthrax every year, select the age of tree to be 6 years 300
The peach tree that growth conditions is similar (kind is honey peach), is randomly divided into three groups, experimental group A, experimental group B and matched group, often group
100.Every peach tree of experimental group A sprays an embodiment 2 before rudiment in spring to tree body multifunctional agricultural (at the beginning of 3 months) is micro-
200 times of diluents of ecological agent A, spend the 200 of the front multifunctional agricultural microbial ecological agent A that tree body sprays an embodiment 2
Times diluent, sprays 200 times of diluents of the multifunctional agricultural microbial ecological agent A of an embodiment 2 to tree body after spending, just fruit
Phase sprays 200 times of diluents of the multifunctional agricultural microbial ecological agent A of an embodiment 2, sprays once real in fruit expanding period
Execute 200 times of diluents of the multifunctional agricultural microbial ecological agent A of example 2.Experimental group B is except by embodiment 2 multi-functional in experimental group A
200 times of diluents of agricultural microbial ecological agent A replace with 200 times of the multifunctional agricultural microbial ecological agent B of the embodiment 2 of equivalent
Outside diluent, other operation is the most identical.Matched group is except by the multifunctional agricultural microbial ecological agent A of embodiment 2 in experimental group A
200 times of diluents replace with outside the sterilized water of equivalent, and other operation is the most identical.Spray 7 days " Invest, Then Investigate " morbidity feelings the last time
Condition, record diseased plant number and state of an illness rank, calculate disease index and preventive effect.Preventive effect %=(matched group disease index-experimental group state of an illness
Index)/matched group disease index × 100, disease index=[∑ (diseased plant numbers at different levels × representative values at different levels)/total strain number × 5] ×
100。
Peach tree anthrax grade scale is as follows:
0 grade: anosis;
1 grade: have a small amount of scab, lesion area accounts for fruit and leaf area less than 1/16;
2 grades: more scab, lesion area accounts for 1/16 to 1/8 (not including 1/16) of fruit and leaf area;
3 grades: lesion area accounts for 1/8 to 1/4 (not including 1/8) of fruit and leaf area;
4 grades: lesion area accounts for 1/4 to 1/2 (not including 1/4) of fruit and leaf area;
5 grades: lesion area accounts for the 1/2 to 3/4 of fruit and leaf area.
The preventive effect of different disposal group peach tree anthrax is as shown in table 1, shows the multifunctional agricultural Tiny ecosystem of embodiment 2
Preparation A reaches 92.39% to the preventive effect of peach tree anthrax, and the multifunctional agricultural microbial ecological agent B of embodiment 2 is to peach tree charcoal
The preventive effect of cellulitis disease reaches 62.24%.
Table 1, the preventive effect of peach tree anthrax
Process | Disease index | Preventive effect |
Matched group | 24.31 | |
Experimental group A | 1.85 | 92.39% |
Experimental group B | 9.18 | 62.24% |
2, the treatment of peach tree anthrax
In a peach garden of the Shandong Province that all can be attacked in various degree by anthrax every year, select the age of tree to be 4 years 90
Growth conditions is similar, the sick level of anthrax is the first fruit phase peach tree (kind is Prunus persicanucipersica Schneider) of 1 grade, is randomly divided into three groups, experimental group
A, experimental group B and matched group, often group 30.The multifunctional agricultural that experimental group A sprayed an embodiment 2 every 7 days to tree body is micro-
200 times of diluents of ecological agent A, spray 5 times altogether.Experimental group B is except by the micro-life of the multifunctional agricultural of embodiment 2 in experimental group A
200 times of diluents of state preparation A replace with 200 times of diluents of the multifunctional agricultural microbial ecological agent B of the embodiment 2 of equivalent
Outward, other operation is the most identical.Matched group is except 200 times of dilutions by the multifunctional agricultural microbial ecological agent A of embodiment in experimental group 2
Liquid replaces with outside the sterilized water of equivalent, and other operation is the most identical.Spray 7 days " Invest, Then Investigate " incidences the last time, calculate disease
Feelings index and curative effect.The computational methods of disease index are with step 1.(before treatment, after disease index-treatment, the state of an illness refers to curative effect %=
Number) front disease index × 100 of/treatment.
The grade scale of peach tree anthrax is with step 1.
The therapeutic effect of different disposal group peach tree anthrax is as shown in table 2, shows the multifunctional agricultural Tiny ecosystem of embodiment 2
Preparation A reaches 83.65% to the therapeutic effect of peach tree anthrax, and the multifunctional agricultural microbial ecological agent B of embodiment 2 is to peach tree charcoal
Cellulitis disease does not has therapeutical effect.
Table 2, the therapeutic effect of peach tree anthrax
Process | Disease index before treatment | Disease index after treatment | Curative effect |
Matched group | 20 | 40.58 | |
Experimental group A | 20 | 3.27 | 83.65% |
Experimental group B | 20 | 26.75 |
It is demonstrated experimentally that the multifunctional agricultural microbial ecological agent A of embodiment 2 can not only the most pre-preventing peach anthrax, also
Peach tree anthrax can be effectively treated.
Embodiment 4, multifunctional agricultural microbial ecological agent promote Camellia sinensis volume increase
Use 300 times of diluents and the multifunctional agricultural of embodiment 2 of the multifunctional agricultural microbial ecological agent A of embodiment 2
300 times of diluents of microbial ecological agent B carry out following experiment.300 times of the multifunctional agricultural microbial ecological agent A of embodiment 2 dilute
Releasing liquid is, with sterilized water, the multifunctional agricultural microbial ecological agent A of embodiment 2 is diluted 300 times of liquid obtained.More than embodiment 2
300 times of diluents of function agricultural microbial ecological agent B are by dilute for the multifunctional agricultural microbial ecological agent B of embodiment 2 with sterilized water
Release 300 times of liquid obtained.
In the tea place of District, Xinyang Area, Henan Province, select above-mentioned Xinyang 10, Fuding DABAI and Pingyang spy early these three XINYANGMAOJIAN product
The Camellia sinensis planted, 90 of the Camellia sinensis that the age of tree is 5 years that each kind equal growth selection state is similar, it is randomly divided into three groups, experiment
Group A, experimental group B and matched group, often group 30.Experimental group A sprays the multi-functional of an embodiment 2 when sprouting sprouts to tree body
300 times of diluents of agricultural microbial ecological agent A, many merits that 10 days spray an embodiment 2 to tree body every time before gathering later
300 times of diluents of the agricultural microbial ecological agent A of energy.Experimental group B is except by the multifunctional agricultural Tiny ecosystem of embodiment 2 in experimental group A
300 times of diluents of preparation A replace with outside 300 times of diluents of multifunctional agricultural microbial ecological agent B of the embodiment 2 of equivalent,
Other operation is the most identical.Matched group is except 300 times of diluents by the multifunctional agricultural microbial ecological agent A of embodiment in experimental group 2
Replacing with outside the sterilized water of equivalent, other operation is the most identical.After Folium Camelliae sinensis is gathered, add up each kind experimental group and matched group
Tea yield, add up rate of growth.Rate of growth=(experimental group yield-matched group yield)/matched group yield.Result such as table 3 institute
Show, show that the multifunctional agricultural microbial ecological agent A of embodiment 2 all has good volume increase to the tea yield of the Camellia sinensis of different cultivars
Effect, average rate of growth is 12%.The Folium Camelliae sinensis of the Camellia sinensis of different cultivars is produced by the multifunctional agricultural microbial ecological agent B of embodiment 2
It is 5% that amount all has effect of increasing production, average rate of growth.The multifunctional agricultural microbial ecological agent A of embodiment 2 to improve tea leaf quality,
Improve millet paste concentration and flavour has good effect.After using, spring tea sprouts early, and bud head is many, holding property good, and harvesting batch is many,
Raising spring tea Famous High-quality Tea ratio and economic benefit are increased extremely advantageous.
Table 3, the Folium Camelliae sinensis rate of growth of each process group
Embodiment 5, multifunctional agricultural microbial ecological agent promote vegetable increase production
The 300 times of diluents using the multifunctional agricultural microbial ecological agent A of embodiment 2 carry out following experiment.Embodiment 2
300 times of diluents of multifunctional agricultural microbial ecological agent A are by the multifunctional agricultural microbial ecological agent A of embodiment 2 with sterilized water
Dilute 300 times of liquid obtained.
The multifunctional agricultural microbial ecological agent A of embodiment 2 is used to enter Fructus Capsici, Semen vignae sinensis and Caulis et Folium Brassicae capitatae in the excellent extraordinary industry of Hebei favour
Row test, all achieves obvious effect of increasing production.
Fructus Capsici test uses randomized block design, is randomly provided 2 treatment regions, and each treatment region sets three repetitions.At 2
Reason district is respectively control treatment district and multifunctional agricultural microbial ecological agent A treatment region (experiment treatment region).The area of each community
It it is 3 mu.Multifunctional agricultural microbial ecological agent A treatment region, after Hot Pepper Seedling transplant survival, sprays an embodiment more than 2 to plant
300 times of diluents of function agricultural microbial ecological agent A, the multifunctional agricultural that plant sprays an embodiment 2 in the setting phase is micro-
300 times of diluents of ecological agent A, spray the multifunctional agricultural microbial ecological agent A's of an embodiment 2 in the best fruiting period to plant
300 times of diluents.Control treatment district is except by micro-for the multifunctional agricultural of embodiment 2 in multifunctional agricultural microbial ecological agent A treatment region
300 times of diluents of ecological agent A replace with outside the sterilized water of equivalent, and other operation is the most identical.After Fructus Capsici is gathered, statistics
Experiment treatment region and the yield of hot pepper in control treatment district, add up rate of growth.Rate of growth=(experiment treatment region yield-control treatment
District's yield)/control treatment district yield.
Result shows, compared with Fructus Capsici control treatment district, and the rate of growth of multifunctional agricultural microbial ecological agent A treatment region Fructus Capsici
It is 20%.
Semen vignae sinensis (kind is asparagus bean) test uses randomized block design, is randomly provided 2 treatment regions, and each treatment region sets
Three repetitions.2 treatment regions are respectively control treatment district and multifunctional agricultural microbial ecological agent A treatment region (experiment treatment region).
The area of each community is 3 mu.Plant is sprayed by multifunctional agricultural microbial ecological agent A treatment region in seedling stage of Semen vignae sinensis once implements
300 times of diluents of the multifunctional agricultural microbial ecological agent A of example 2, enter seat pod after date at Semen vignae sinensis and plant spray twice enforcement
300 times of diluents of the multifunctional agricultural microbial ecological agent A of example 2, this is spaced 10 days between spraying for twice.Control treatment district removes will
In multifunctional agricultural microbial ecological agent A treatment region, 300 times of diluents of the multifunctional agricultural microbial ecological agent A of embodiment 2 are replaced
Outside for the sterilized water of equivalent, other operation is the most identical.After Semen vignae sinensis is gathered, statistical experiment treatment region and the cowpea in control treatment district
Bean yield, adds up rate of growth.Rate of growth=(experiment treatment region yield-control treatment district yield)/control treatment district yield.
Result shows, compared with control treatment district, multifunctional agricultural microbial ecological agent A treatment region Semen vignae sinensis rate of growth is
15%.Multifunctional agricultural microbial ecological agent A can delay Semen vignae sinensis old and feeble, increases the yield of the fresh Semen vignae sinensis of secondary recovery.
Caulis et Folium Brassicae capitatae (in kind sweet No. 11) test uses randomized block design, is randomly provided 2 treatment regions, and each treatment region sets
Three repetitions.2 treatment regions are respectively control treatment district and multifunctional agricultural microbial ecological agent A treatment region (experiment treatment region).
The area of each community is 1 mu.Plant, after cabbage seedling transplant survival, is sprayed by multifunctional agricultural microbial ecological agent A treatment region
300 times of diluents of the multifunctional agricultural microbial ecological agent A of embodiment 2, spray an embodiment 2 in the rosette state to plant
300 times of diluents of multifunctional agricultural microbial ecological agent A, in the heading stage plant sprayed the multi-functional agriculture of an embodiment 2
With 300 times of diluents of microbial ecological agent A.Control treatment district is except by embodiment 2 in multifunctional agricultural microbial ecological agent A treatment region
300 times of diluents of multifunctional agricultural microbial ecological agent A replace with outside the sterilized water of equivalent, other operation is the most identical.Result
Show the cabbage head diameter big 4-of cabbage head diameter than comparison treatment region of multifunctional agricultural microbial ecological agent A treatment region
6cm.Leaf is multiple, Ye great, leaf thick, the effect of Bao Xin great to illustrate have rush to leaf vegetables by multifunctional agricultural microbial ecological agent A.
It is demonstrated experimentally that enzyme system material in crop is synthesized by the multifunctional agricultural microbial ecological agent A of embodiment 2, activity increases
Add and all have regulation effect, crop is had growth-promoting degeneration-resistant, immunity in strong sprout, spend more, many real, many, improve yield, improve quality it
Effect.
Claims (10)
- The most agricultural complex micro organism fungicide, its active component is by A Yebo plurispore bacillus, Bacillus licheniformis, Lyceum spore Bacillus and east acetic acid bacteria composition.
- Agricultural complex micro organism fungicide the most according to claim 1, it is characterised in that: described A Yebo plurispore bacillus is A Yebo plurispore bacillus ACCC 19947, described Bacillus licheniformis is Bacillus licheniformis ACCC 19946, described Lyceum Bacillus cereus is Lyceum bacillus cereus ACCC 19948, and described east acetic acid bacteria is east acetic acid bacteria ACCC 19949.
- Agricultural complex micro organism fungicide the most according to claim 1 and 2, it is characterised in that: described agricultural complex microorganism In microbial inoculum, the colony-forming units ratio of A Yebo plurispore bacillus, Bacillus licheniformis, Lyceum bacillus cereus and east acetic acid bacteria For (2.5-4): 1:(1-2): (3-4.5).
- 4. according to the agricultural complex micro organism fungicide described in claim 1 or 2 or 3, it is characterised in that: described agricultural composite microbial In thing microbial inoculum, A Yebo plurispore bacillus, Bacillus licheniformis, Lyceum bacillus cereus and the colony-forming units of east acetic acid bacteria Ratio is 3.25:1:1.5:3.75.
- 5. arbitrary described agricultural compound in arbitrary described agricultural complex micro organism fungicide or claim 1-4 in claim 1-4 Under described A Yebo plurispore bacillus, Bacillus licheniformis, Lyceum bacillus cereus and east acetic acid bacteria in microbial bacterial agent State any one to apply:1) application in preparation plant Anthracnose Pathogen bacteria inhibitor;2) application in prevention and/or treatment plant anthrax;3) application in promoting crop yield;4) application in arbitrary described agricultural microbial ecological agent in preparation claim 7-9.
- Application the most according to claim 5, it is characterised in that: described plant is rosaceous plant, and described crop is Theaceae Crop, solanaceous crops, legume or crop in cruciferae.
- The most agricultural microbial ecological agent, its active component is to cultivate in claim 1-4 arbitrary described agricultural with fermentation medium The culture that complex micro organism fungicide obtains.
- Agricultural microbial ecological agent the most according to claim 7, it is characterised in that: in described agricultural microbial ecological agent, described The colony-forming units of A Yebo plurispore bacillus, Bacillus licheniformis, Lyceum bacillus cereus and east acetic acid bacteria is than being (10- 30): 1:(1-5): (30-50) or 18:1:1.6:40.
- 9. any one application following of the agricultural microbial ecological agent described in claim 7 or 8:1) application in preparation plant Anthracnose Pathogen bacteria inhibitor;2) application in prevention and/or treatment plant anthrax;3) application in promoting crop yield.
- Application the most according to claim 9, it is characterised in that: described plant is rosaceous plant, and described crop is Theaceae Crop, solanaceous crops, legume or crop in cruciferae.
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