CN110477023A - The biological agent and preparation method thereof for preventing and treating powdery mildew of strawberry - Google Patents
The biological agent and preparation method thereof for preventing and treating powdery mildew of strawberry Download PDFInfo
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- CN110477023A CN110477023A CN201910997697.7A CN201910997697A CN110477023A CN 110477023 A CN110477023 A CN 110477023A CN 201910997697 A CN201910997697 A CN 201910997697A CN 110477023 A CN110477023 A CN 110477023A
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- powdery mildew
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/08—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing solids as carriers or diluents
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/12—Powders or granules
- A01N25/14—Powders or granules wettable
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N61/00—Biocides, pest repellants or attractants, or plant growth regulators containing substances of unknown or undetermined composition, e.g. substances characterised only by the mode of action
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
Abstract
The present invention relates to a kind of biological agents and preparation method thereof for preventing and treating powdery mildew of strawberry, belong to disease control field.The biological agent is mainly grouped as by Harpin albumen, fermentation of bacillus subtilis bacterium solution and absorption carrier group, can not only prevention & protection powdery mildew of strawberry generation, and can cure protection powdery mildew of strawberry.Wherein component Harpin albumen can make plant generate disease-resistant, the degeneration-resistant effect of insect pest conjunction, the active material generated during bacillus subtilis thalli growth has apparent inhibiting effect to the conditioned pathogen of pathogenic bacteria or autogenous infection, conducive to the prevention and improvement of powdery mildew of strawberry.In addition, the biological agent can provide nutrition for crop, crop nutrient growth and reproductive growth are automatically adjusted, improves crop yield.Component Harpin albumen in the biological agent is a kind of highly resistance immune protein, nutrition can be provided for crop, and can promote plant growth, conducive to the raising of crop yield.
Description
Technical field
The present invention relates to a kind of biological agents and preparation method thereof for preventing and treating powdery mildew of strawberry, belong to disease prevention techniques
Field.
Background technique
Powdery mildew of strawberry is the environment that the common important disease of strawberry, especially strawberry are in low temperature and high relative humidity, powdery mildew
Incidence probability is very big, is generally possible to cause the strawberry field underproduction 20%~30%, severe one can the underproduction 50%, or even total crop failure seriously affects
The planting benefit at strawberry cultivating families.
Currently, the measure of prevention and treatment powdery mildew of strawberry has cultural control, mainly includes efficent rotation, selection disease-resistant variety, closes
Reason fertilising is rationally poured water, and main control measure is chemical prevention, based on the progress of chemical pesticide fungicide afterwards prevention and treatment, but
The a wide range of use of agriculture chemicals fungicide causes strawberry, Pesticide-Polluted Soil exceeded, affects the quality of strawberry, makes to food safety
At threat, also it is unfavorable for the long-term sustainable development of soil.
In view of above-mentioned status, it is very active to become current plant pathology educational circles using antagonistic microbe controlling plant diseases
One of research field.Applicant provides by studying for many years in terms of antagonistic microbe prevents and treats and utilizes high quick albumen and micro- life
The microbial pesticide that object collective effect is constituted, can effectively prevent powdery mildew of strawberry, can further nutrition plant, increase strawberry
It produces, and without side-effects to plant and soil.
Existing there are still agriculture chemicals fungicides to cause plant and Pesticide-Polluted Soil exceeded, and food safety is low, is unfavorable for soil
The deficiency of long-run development researches and develops a adjusting plant vegetative growth and reproductive growth hereby, increases the prevention and treatment strawberry of crop yield
Biological agent of powdery mildew and preparation method thereof is vital.
Summary of the invention
The technical problem to be solved by the present invention is overcoming the deficiencies in the prior art, it is white to provide a kind of prevention and treatment strawberry
The biological agent and preparation method thereof of powder disease.
The technical scheme to solve the above technical problems is that
It is a kind of prevent and treat powdery mildew of strawberry biological agent, mainly by Harpin albumen, bacillus subtilis bacillus fermentation bacterium solution and
Absorption carrier group is grouped as.
Preferably, the weight proportion of the component is protein 10~15 part Harpin, fermentation of bacillus subtilis bacterium solution 40
~60 parts, 20~30 parts of absorption carrier.
Preferably, the absorption carrier is precipitated calcium carbonate, seawood meal, diatomite, kaolin, bentonite or atlapulgite
At least one of.
A kind of preparation method of biological agent that preventing and treating powdery mildew of strawberry, includes the following steps,
Step 1 chooses Bacillus subtilis strain, activated, expansion culture and fermentation, obtains withered grass gemma after cell cracking
Bacillus fermentation bacterium solution；
Step 2 weighs Harpin albumen, fermentation of bacillus subtilis bacterium solution and absorption carrier according to formula, is uniformly mixed
Adsorption precipitation stands 0.5~1.0h, obtains mixing suspension；
Step 3 carries out low temperature spray drying by above-mentioned mixing suspension under conditions of 80 DEG C~120 DEG C, obtains drying
Primary biological agent；
Above-mentioned primary biological agent is carried out crushing operation by step 4, until the granularity of powderous preparations is less than 100 mesh, it is as biological
Formulation products；
Above-mentioned biological agent product is carried out packed in flexible pouchy by step 5.
Preferably, the actication of culture in the step 1 uses solid plate culture, the culture that solid plate culture uses
Base is PDA solid medium, and PDA solid medium includes that 200g potato, 20g glucose or sucrose make raw material, by potato
30min filtering is boiled in water, takes and 20g glucose or sucrose and 20g agar is added in filtered supernatant, and adds water fixed
Hold to 1000mL, 121 DEG C~125 DEG C of temperature, disinfecting action 0.5h is carried out under the conditions of the MPa of pressure 0.103MPa~0.168, and
24~48h is cultivated in 35 DEG C of constant incubator.
Preferably, the strain in the step 1 expands what culture was used using shaking flask Liquid Culture, shaking flask Liquid Culture
Culture medium is PDA liquid medium, and PDA liquid medium includes that 200g potato and 20g glucose or sucrose make raw material, will
Potato boils 30min filtering in water, 20g glucose or sucrose is added in filtered supernatant, and add water constant volume
To 1000mL, 121 DEG C~125 DEG C of Yu Wendu, disinfecting action 0.5h is carried out under the conditions of the MPa of pressure 0.103MPa~0.168, then
It is inoculated into PDA fluid nutrient medium from picking thallus on above-mentioned PDA solid medium, in revolving speed 200rpm~225rpm, temperature
24~48h of shaken cultivation under the conditions of 35 DEG C of degree, obtains fermentation seed liquid, and the inoculum concentration is 2%.
Potassium dihydrogen phosphate and 1000ml water, pH are 7.2~7.4, when spore forming rate reaches 80%, thallus fermentation process
It completes, stops fermentation.Preferably, the fermentation in the step 1 uses fermentation tank culture, by fermentation seed liquid with 2% inoculum concentration
It goes in fermentor and ferments, first go out under the conditions of 121 DEG C~125 DEG C of temperature, pressure 0.103MPa~0.168 MPa
Bacterium handle 1.0h, after ferment under the conditions of 35 DEG C of temperature, revolving speed 200rpm, ventilatory capacity 1:1, the fermentation tank culture medium includes
10g peptone, 5g yeast extract, 3.5g glucose, 5g sodium chloride, 0.5g
Preferably, the cell cracking in the step 1 is to the bacillus thallus after fermentation tank culture at 75 DEG C of temperature
15s~16s is heated at~90 DEG C, carries out high temperature, short time cracking, to be crushed somatic cells, obtains zymocyte liquid.
Preferably, the spray drying in the step 3 is carried out using spray dryer, and air inlet is required in spray dryer
Temperature is 150 DEG C~160 DEG C, intake 40000m3/ h~45000m3/ h, temperature of outgoing air are 70 DEG C~80 DEG C, and exhaust air rate is
40000m3/ h~45000m3/ h, mixing suspension through high speed centrifugal atomization device, rotating spraying at imperceptible misty liquid droplets, with
Hot-air parallel flow contact, rapid draing powdering object obtain dry primary biological agent.
Preferably, the biological agent of the prevention and treatment powdery mildew of strawberry is white in prevention and treatment powdery mildew of strawberry, uncinula necator, cucumber
The application of powder disease, watermelon powdery mildew disease or melon powdery mildew.
Preferably, the biological agent of the prevention and treatment powdery mildew of strawberry is diluted with water 600~1000 times, is sprayed on crop blade face
Application upper or that pouring root, leaching root or seed soaking are carried out to crop.
The beneficial effects of the present invention are: 1, disease control, the biological agent can not only prevention & protection powdery mildew of strawberry hair
It is raw, there is disease-resistant function, and protection powdery mildew of strawberry, the ability for having auxiliary to cure the disease can be cured.Component in the biological agent
Harpin albumen can make plant generate disease-resistant, the degeneration-resistant effect of insect pest conjunction, generate during bacillus subtilis thalli growth
Subtilin, polymyxins, nystatin, gramicidins isoreactivity substance are to the conditioned pathogens of pathogenic bacteria or autogenous infection
There is apparent inhibiting effect, conducive to the prevention and improvement of powdery mildew of strawberry；2, promote growth, increase crop yield, the biology system
Agent can provide nutrition for crop, automatically adjust crop nutrient growth and reproductive growth, improve the yield of crop.The biology system
Component Harpin albumen in agent is a kind of highly resistance immune protein, and nutrition can be provided for crop, and having can promote plant raw
Long effect, conducive to the raising of crop yield.
With embodiment
The principles and features of the present invention are described below, and the given examples are served only to explain the present invention, is not intended to limit this
The range of invention.
A kind of biological agent for preventing and treating powdery mildew of strawberry, mainly by Harpin albumen, bacillus subtilis bacillus fermentation bacterium
Liquid and absorption carrier group are grouped as.It is compounded by Harpin albumen with bacillus subtilis bacterium solution, in conjunction with the advantage of the two, not only
The generation that can prevent powdery mildew of strawberry has disease-resistant function, and can cure powdery mildew of strawberry, the ability for having auxiliary to cure the disease.Together
When, Harpin albumen can provide nutrition for crop, promote plant growth, improve crop yield, realize what a medicine was mostly used
The weight proportion unit of effect, the component in the application is used uniformly kg.
The weight proportion of the component be protein 10~15 part Harpin, 40~60 parts of fermentation of bacillus subtilis bacterium solution,
20~30 parts of absorption carrier.
The absorption carrier be precipitated calcium carbonate, seawood meal, diatomite, kaolin, bentonite or atlapulgite in extremely
Few one kind.
A kind of preparation method of biological agent that preventing and treating powdery mildew of strawberry, includes the following steps,
Step 1 chooses Bacillus subtilis strain, activated, expansion culture and fermentation, obtains withered grass gemma after cell cracking
Bacillus fermentation bacterium solution；
Step 2 weighs Harpin albumen, fermentation of bacillus subtilis bacterium solution and absorption carrier according to formula, is uniformly mixed
Adsorption precipitation stands 0.5~1.0h, obtains mixing suspension；
Step 3 carries out low temperature spray drying by above-mentioned mixing suspension under conditions of 80 DEG C~120 DEG C, obtains drying
Primary biological agent；
Above-mentioned primary biological agent is carried out crushing operation by step 4, until the granularity of powderous preparations is less than 100 mesh, it is as biological
Formulation products；
Above-mentioned biological agent product is carried out packed in flexible pouchy by step 5.
Actication of culture in the step 1 uses solid plate culture, and the culture medium that solid plate culture uses is PDA
Solid medium, PDA solid medium include that 200g potato, 20g glucose or sucrose make raw material, in water by potato
30min filtering is boiled, takes and 20g glucose or sucrose and 20g agar is added in filtered supernatant, and water is added to be settled to
1000mL, disinfecting action 0.5h is carried out under the conditions of the MPa of pressure 0.103MPa~0.168, and 35 by 121 DEG C~125 DEG C of temperature
DEG C constant incubator in cultivate 24~48h.
Strain in the step 1 expands culture and uses shaking flask Liquid Culture, the culture medium that shaking flask Liquid Culture uses for
PDA liquid medium, PDA liquid medium include that 200g potato and 20g glucose or sucrose make raw material, and potato is existed
30min filtering is boiled in water, 20g glucose or sucrose is added in filtered supernatant, and water is added to be settled to 1000mL,
Disinfecting action 0.5h is carried out under the conditions of 121 DEG C~125 DEG C of temperature, the MPa of pressure 0.103MPa~0.168, then from above-mentioned PDA
Picking thallus is inoculated into PDA fluid nutrient medium on solid medium, in 35 DEG C of revolving speed 200rpm~225rpm, temperature conditions
24~48h of lower shaken cultivation, obtains fermentation seed liquid, and the inoculum concentration is 2%.
Under the conditions of MPa carry out sterilization treatment 1.0h, after issued in 35 DEG C of temperature, revolving speed 200rpm, ventilatory capacity 1:1 condition
Ferment, the fermentation tank culture medium include 10g peptone, 5g yeast extract, 3.5g glucose, 5g sodium chloride, step 1 described in 0.5g
In fermentation use fermentation tank culture, fermentation seed liquid is gone in fermentor with 2% inoculum concentration and is fermented, first in temperature 121
DEG C~125 DEG C, the potassium dihydrogen phosphate of pressure 0.103MPa~0.168 and 1000ml water, pH is 7.2~7.4, works as spore forming rate
When reaching 80%, thallus fermentation process is completed, and stops fermentation.
Cell cracking in the step 1 be to the bacillus thallus after fermentation tank culture 75 DEG C of temperature~
15s~16s is heated at 90 DEG C, carries out high temperature, short time cracking, to be crushed somatic cells, obtains zymocyte liquid.
Spray drying in the step 3 is dried using spray dryer, and inlet air temperature is required in spray dryer
It is 150 DEG C~160 DEG C, intake 40000m3/ h~45000m3/ h, temperature of outgoing air are 70 DEG C~80 DEG C, and exhaust air rate is
40000m3/ h~45000m3/ h, mixing suspension through high speed centrifugal atomization device, rotating spraying at imperceptible misty liquid droplets, with
Hot-air parallel flow contact, rapid draing powdering object obtain dry primary biological agent.
The biological agent of the prevention and treatment powdery mildew of strawberry is in prevention and treatment powdery mildew of strawberry, uncinula necator, powdery mildew of cucumber, west
The application of melon powdery mildew disease or melon powdery mildew.
The biological agent of the prevention and treatment powdery mildew of strawberry is diluted with water 600~1000 times, and it is on crop blade face or right to be sprayed on
Crop carries out the application of pouring root, leaching root or seed soaking.
Embodiment 1
A kind of biological agent for preventing and treating powdery mildew of strawberry, mainly by Harpin protein 10 kg, bacillus subtilis bacillus fermentation bacterium
Liquid 50kg and precipitated calcium carbonate 25kg group are grouped as.
A kind of preparation method of biological agent that preventing and treating powdery mildew of strawberry, includes the following steps,
Step 1 chooses Bacillus subtilis strain, activated, expansion culture and fermentation, obtains withered grass gemma after cell cracking
Bacillus fermentation bacterium solution；
The actication of culture uses solid plate culture, and the culture medium that solid plate culture uses is PDA solid medium, PDA
Solid medium includes that 200g potato, 20g glucose or sucrose make raw material, and potato is boiled to 30min filtering in water,
It takes and 20g glucose or sucrose and 20g agar is added in filtered supernatant, and water is added to be settled to 1000mL, 123 DEG C of temperature,
Disinfecting action 0.5h is carried out under the conditions of pressure 0.132MPa, and cultivates 36h in 35 DEG C of constant incubator；
The strain expands culture and uses shaking flask Liquid Culture, the culture medium that shaking flask Liquid Culture uses for PDA liquid medium,
PDA liquid medium includes that 200g potato and 20g glucose or sucrose make raw material, and potato is boiled 30min in water
20g glucose or sucrose is added in filtering in filtered supernatant, and water is added to be settled to 1000mL, 123 DEG C of Yu Wendu, presses
Disinfecting action 0.5h is carried out under the conditions of strong 0.132 MPa, then is inoculated into PDA from picking thallus on above-mentioned PDA solid medium
In fluid nutrient medium, shaken cultivation 36h under the conditions of revolving speed 212rpm, 35 DEG C of temperature obtains fermentation seed liquid, the inoculum concentration
It is 2%；
The fermentation uses fermentation tank culture, and fermentation seed liquid is gone in fermentor with 2% inoculum concentration and is fermented, first in temperature
123 DEG C of degree carries out sterilization treatment 1.0h under the conditions of pressure 0.132MPa, after in 35 DEG C of temperature, revolving speed 200rpm, ventilatory capacity 1:1
Under the conditions of ferment, the fermentation tank culture medium includes 10g peptone, 5g yeast extract, 3.5g glucose, 5g sodium chloride, 0.5g phosphorus
Acid dihydride potassium and 1000ml water, pH 7.3, when spore forming rate reaches 80%, thallus fermentation process is completed, and stops fermentation；
The cell cracking is to heat 15.5s at 80 DEG C of temperature to the bacillus thallus after fermentation tank culture, is carried out
High temperature, short time cracking obtains zymocyte liquid to be crushed somatic cells；
Step 2 weighs Harpin protein 10 kg, fermentation of bacillus subtilis bacterium solution 50kg and precipitated calcium carbonate according to formula
25kg is uniformly mixed adsorption precipitation, stands 0.8h, and precipitated calcium carbonate is sufficiently enriched with, and obtains mixing suspension；
Step 3 carries out low temperature spray drying, requires in spray dryer by above-mentioned mixing suspension under conditions of 100 DEG C
Inlet air temperature is 155 DEG C, intake 43000m3/ h, temperature of outgoing air are 75 DEG C, exhaust air rate 43000m3/ h, mixing suspension
Through high speed centrifugal atomization device, rotating spraying is at imperceptible misty liquid droplets, with hot-air parallel flow contact, rapid draing powdering
Object obtains dry primary biological agent；
Above-mentioned primary biological agent is carried out crushing operation by step 4, until the granularity of powderous preparations is less than 100 mesh, it is as biological
Formulation products；
Above-mentioned biological agent product is carried out packed in flexible pouchy by step 5.
Embodiment 2
A kind of biological agent for preventing and treating powdery mildew of strawberry, mainly by Harpin protein 12 kg, bacillus subtilis bacillus fermentation bacterium
Liquid 60kg and precipitated calcium carbonate 30kg group are grouped as.
A kind of preparation method of biological agent that preventing and treating powdery mildew of strawberry, includes the following steps,
Step 1 chooses Bacillus subtilis strain, activated, expansion culture and fermentation, obtains withered grass gemma after cell cracking
Bacillus fermentation bacterium solution；
The actication of culture uses solid plate culture, and the culture medium that solid plate culture uses is PDA solid medium, PDA
Solid medium includes that 200g potato, 20g glucose or sucrose make raw material, and potato is boiled to 30min filtering in water,
It takes and 20g glucose or sucrose and 20g agar is added in filtered supernatant, and water is added to be settled to 1000mL, 125 DEG C of temperature,
Disinfecting action 0.5h is carried out under the conditions of pressure 0.168MPa, and cultivates 48h in 35 DEG C of constant incubator；
The strain expands culture and uses shaking flask Liquid Culture, the culture medium that shaking flask Liquid Culture uses for PDA liquid medium,
PDA liquid medium includes that 200g potato and 20g glucose or sucrose make raw material, and potato is boiled 30min in water
20g glucose or sucrose is added in filtering in filtered supernatant, and water is added to be settled to 1000mL, 125 DEG C of Yu Wendu, presses
Disinfecting action 0.5h is carried out under the conditions of strong 0.168 MPa, then is inoculated into PDA from picking thallus on above-mentioned PDA solid medium
In fluid nutrient medium, shaken cultivation 48h under the conditions of revolving speed 225rpm, 35 DEG C of temperature obtains fermentation seed liquid, the inoculum concentration
It is 2%；
The fermentation uses fermentation tank culture, and fermentation seed liquid is gone in fermentor with 2% inoculum concentration and is fermented, first in temperature
125 DEG C of degree carries out sterilization treatment 1.0h under the conditions of pressure 0.168MPa, after in 35 DEG C of temperature, revolving speed 200rpm, ventilatory capacity 1:1
Under the conditions of ferment, the fermentation tank culture medium includes 10g peptone, 5g yeast extract, 3.5g glucose, 5g sodium chloride, 0.5g phosphorus
Acid dihydride potassium and 1000ml water, pH 7.4, when spore forming rate reaches 80%, thallus fermentation process is completed, and stops fermentation；
The cell cracking is to heat 16s at 90 DEG C of temperature to the bacillus thallus after fermentation tank culture, is carried out high
Temperature cracks in short-term, to be crushed somatic cells, obtains zymocyte liquid；
Step 2 weighs Harpin protein 12 kg, fermentation of bacillus subtilis bacterium solution 60kg and precipitated calcium carbonate according to formula
30kg is uniformly mixed adsorption precipitation, stands 0.1h, and precipitated calcium carbonate is sufficiently enriched with, and obtains mixing suspension；
Step 3 carries out low temperature spray drying, requires in spray dryer by above-mentioned mixing suspension under conditions of 120 DEG C
Inlet air temperature is 160 DEG C, intake 45000m3/ h, temperature of outgoing air are 80 DEG C, exhaust air rate 45000m3/ h, mixing suspension
Through high speed centrifugal atomization device, rotating spraying is at imperceptible misty liquid droplets, with hot-air parallel flow contact, rapid draing powdering
Object obtains dry primary biological agent；
Above-mentioned primary biological agent is carried out crushing operation by step 4, until the granularity of powderous preparations is less than 100 mesh, it is as biological
Formulation products；
Above-mentioned biological agent product is carried out packed in flexible pouchy by step 5.
Embodiment 3
A kind of biological agent for preventing and treating powdery mildew of strawberry, mainly by Harpin protein 15 kg, bacillus subtilis bacillus fermentation bacterium
Liquid 40kg and precipitated calcium carbonate 20kg group are grouped as.
A kind of preparation method of biological agent that preventing and treating powdery mildew of strawberry, includes the following steps,
Step 1 chooses Bacillus subtilis strain, activated, expansion culture and fermentation, obtains withered grass gemma after cell cracking
Bacillus fermentation bacterium solution；
The actication of culture uses solid plate culture, and the culture medium that solid plate culture uses is PDA solid medium, PDA
Solid medium includes that 200g potato, 20g glucose or sucrose make raw material, and potato is boiled to 30min filtering in water,
It takes and 20g glucose or sucrose and 20g agar is added in filtered supernatant, and water is added to be settled to 1000mL, 121 DEG C of temperature,
Disinfecting action 0.5h is carried out under the conditions of pressure 0.103MPa, and is cultivated for 24 hours in 35 DEG C of constant incubator；
The strain expands culture and uses shaking flask Liquid Culture, the culture medium that shaking flask Liquid Culture uses for PDA liquid medium,
PDA liquid medium includes that 200g potato and 20g glucose or sucrose make raw material, and potato is boiled 30min in water
20g glucose or sucrose is added in filtering in filtered supernatant, and water is added to be settled to 1000mL, 121 DEG C of Yu Wendu, presses
Disinfecting action 0.5h is carried out under the conditions of strong 0.103MPa, then is inoculated into PDA liquid from picking thallus on above-mentioned PDA solid medium
In body culture medium, shaken cultivation for 24 hours, obtains fermentation seed liquid, the inoculum concentration is under the conditions of revolving speed 200rpm, 35 DEG C of temperature
2%；
The fermentation uses fermentation tank culture, and fermentation seed liquid is gone in fermentor with 2% inoculum concentration and is fermented, first in temperature
121 DEG C of degree carries out sterilization treatment 1.0h under the conditions of pressure 0.103MPa, after in 35 DEG C of temperature, revolving speed 200rpm, ventilatory capacity 1:1
Under the conditions of ferment, the fermentation tank culture medium includes 10g peptone, 5g yeast extract, 3.5g glucose, 5g sodium chloride, 0.5g phosphorus
Acid dihydride potassium and 1000ml water, pH 7.2, when spore forming rate reaches 80%, thallus fermentation process is completed, and stops fermentation；
The cell cracking is to heat 15s at 75 DEG C of temperature to the bacillus thallus after fermentation tank culture, is carried out high
Temperature cracks in short-term, to be crushed somatic cells, obtains zymocyte liquid；
Step 2 weighs Harpin protein 15 kg, fermentation of bacillus subtilis bacterium solution 40kg and precipitated calcium carbonate according to formula
20kg is uniformly mixed adsorption precipitation, stands 0.5h, and precipitated calcium carbonate is sufficiently enriched with, and obtains mixing suspension；
Step 3 carries out low temperature spray drying by above-mentioned mixing suspension under conditions of 80 DEG C, require in spray dryer into
Air temperature is 150 DEG C, intake 40000m3/ h, temperature of outgoing air are 70 DEG C, exhaust air rate 40000m3/ h, mixing suspension warp
High speed centrifugal atomization device, rotating spraying is at imperceptible misty liquid droplets, and hot-air parallel flow contact, rapid draing powdering object,
Obtain dry primary biological agent；
Above-mentioned primary biological agent is carried out crushing operation by step 4, until the granularity of powderous preparations is less than 100 mesh, it is as biological
Formulation products；
Above-mentioned biological agent product is carried out packed in flexible pouchy by step 5.
The biological agent of the prevention and treatment powdery mildew of strawberry of the present embodiment is in use, be diluted with water 600~1000 for biological agent
Again, it is sprayed on crop blade face after；Interval sprays 1 time for 7~10 days, and the prevention and treatment to crop powdery mildew can be realized, such as arid,
Low temperature, etc. the disastrous time, can optionally add 1~2 time and spray.The medical fluid after dilution can also be subjected to pouring root, leaching to crop
Root and seed soaking can also prevent crop powdery mildew.
Embodiment 4
It is right to examine the biological agent of prevention and treatment powdery mildew of strawberry of the invention in the control efficiency of powdery mildew of strawberry and increasing production function
The biological agent of prevention and treatment powdery mildew of strawberry and existing routine administration, clear water obtained by 1-3 of the embodiment of the present invention compare and analyze examination
It tests.
Test period: in September, 2017~2018 year May；Test site: Yantai City bar town town Zhu Liu Wang Cun.
For trying soil: soil types is brown earth, and irrigation condition is good.0-20cm soil sample is taken to chemically examine before test, soil is supported
Divide situation such as table 1.
1 experimental plot soil nutrient status table of table
| Time | Place | Organic matter (g/kg) | Alkali-hydrolyzable nitrogen (mg/kg) | Rapid available phosphorus (mg/kg) | Available potassium (mg/kg) | PH value |
| 2017 | Zhu Liu Wang Cun | 12.0 | 80.5 | 62.0 | 110.0 | 5.5 |
For examination crop varieties and planting type: for studying object strawberry, kind sweet tea is precious, is colonized within September 10th, 0.15 m of spacing in the rows, row spacing
0.75m, the high 0.40m in ridge, the long 9m in ridge, ridge face width 0.6 m, furrow 0.2m, double line width ridge, mu plant 11850 plants.
Test method: test sets 5 processing, 1,600 times of dilutions of biological agent of the prevention and treatment powdery mildew of strawberry of embodiment 1；
2, embodiment 2 prevents and treats 800 times of dilutions of biological agent of powdery mildew of strawberry；3, embodiment 3 prevents and treats the biology system of powdery mildew of strawberry
1000 times of dilutions of agent；4,50% carbendazim, 800 times of dilutions；5, clear water.The plant for selecting growing way similar with morbidity, selects altogether
5000 plants of strawberries are divided into five groups, every group 1000 plants, are handled respectively with above-mentioned 5 reagents the strawberry with powdery mildew.In
Powdery mildew of strawberry originates the phase and sprays for the first time, sprays every 7 days primary, and three times, each processing group is according to spraying 3kg for continuous spraying
Standard carry out.
Preventive effect calculates: 0 grade: disease-free；1 grade: lesion area accounts for 5% or less blade area；3 grades: lesion area accounts for blade face
Long-pending 6%-10%；5 grades: lesion area accounts for the 11%-25% of blade area；7 grades: lesion area accounts for the 26%-50% of blade area；9
Grade: lesion area accounts for 51% of blade area or more.Disease index and control efficiency is calculated as follows:
Disease index=∑ (the disease numbers of sheets at different levels × opposite value of series)/(investigation total number of sheets × 9) × 100
Control efficiency (%)=(control disease index-processing disease index)/control disease index × 100%
As a result with analysis
Influence of the different disposal reagent to strawberry biological character
Field observation, processing group 1-3 spray the strawberry of biological agent compared with the strawberry that other processing groups spray, and strawberry leaf color is dark green to be had
Gloss, growing way is vigorous, and powdery mildew cure rate is higher, and specific control efficiency is referring to the following table 2.
Table 2 prevents and treats prevention and treatment efficiency table of the biological agent to powdery mildew of strawberry of powdery mildew of strawberry
Conclusion (of pressure testing)
By data analysis in upper table, it is found that three times after spray, the biological agent that embodiment 1-3 prevents and treats powdery mildew of strawberry is white to strawberry
The prevention and treatment efficiency of powder disease reaches as high as 92.67%, and is superior to the 800 times of dilution medications of traditional 50% carbendazim, prevents and treats efficiency
It is 82.13%.It can be seen that the biological agent of present invention prevention and treatment powdery mildew of strawberry is sprayed, it is aobvious to the control efficiency of powdery mildew of strawberry
Enhancing is write, powdery mildew of strawberry disease incidence greatly reduces, thus, the present invention prevents and treats the biological agent of powdery mildew of strawberry in prevention and treatment strawberry
There is splendid control efficiency in terms of powdery mildew.
Influence of the above-mentioned processing group processing operation of statistic mixed-state to yield of strawberry.
Fruit started to harvest in 14 days 2 months, and each cell list is adopted, list receives cumulative production, and May 25, harvesting terminated.Yield knot
Fruit is shown in Table 3.
Table 3 prevents and treats influence table of the biological agent of powdery mildew of strawberry to yield of strawberry
Yield of strawberry result table
The foregoing is merely presently preferred embodiments of the present invention, is not intended to limit the invention, all in the spirit and principles in the present invention
Within, any modification, equivalent replacement, improvement and so on should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of biological agent for preventing and treating powdery mildew of strawberry, it is characterised in that: mainly sent out by Harpin albumen, bacillus subtilis
Yeast-like fungi liquid and absorption carrier group are grouped as.
2. the biological agent of prevention and treatment powdery mildew of strawberry according to claim 1, it is characterised in that: the weight of the component is matched
Than for protein 10~15 part Harpin, 40~60 parts of fermentation of bacillus subtilis bacterium solution, 20~30 parts of absorption carrier.
3. the biological agent of prevention and treatment powdery mildew of strawberry according to claim 1, it is characterised in that: the absorption carrier is light
At least one of matter calcium carbonate, seawood meal, diatomite, kaolin, bentonite or atlapulgite.
4. a kind of preparation method for the biological agent for preventing and treating powdery mildew of strawberry, it is characterised in that: include the following steps,
Step 1 chooses Bacillus subtilis strain, activated, expansion culture and fermentation, obtains withered grass gemma after cell cracking
Bacillus fermentation bacterium solution；
Step 2 weighs Harpin albumen, fermentation of bacillus subtilis bacterium solution and absorption carrier according to formula, is uniformly mixed
Adsorption precipitation stands 0.5~1.0h, obtains mixing suspension；
Step 3 carries out low temperature spray drying by above-mentioned mixing suspension under conditions of 80 DEG C~120 DEG C, obtains drying
Primary biological agent；
Above-mentioned primary biological agent is carried out crushing operation by step 4, until the granularity of powderous preparations is less than 100 mesh, it is as biological
Formulation products；
Above-mentioned biological agent product is carried out packed in flexible pouchy by step 5.
5. the preparation method of the biological agent of prevention and treatment powdery mildew of strawberry according to claim 4, it is characterised in that: the step
Actication of culture in rapid one uses solid plate culture, and the culture medium that solid plate culture uses is PDA solid medium, PDA
Solid medium includes that 200g potato, 20g glucose or sucrose make raw material, and potato is boiled to 30min filtering in water,
It takes and 20g glucose or sucrose and 20g agar is added in filtered supernatant, and water is added to be settled to 1000mL, 121 DEG C of temperature
~125 DEG C, disinfecting action 0.5h is carried out under the conditions of the MPa of pressure 0.103MPa~0.168, and in 35 DEG C of constant incubator
Cultivate 24~48h.
6. the preparation method of the biological agent of prevention and treatment powdery mildew of strawberry according to claim 4, it is characterised in that: the step
Strain in rapid one expands culture and uses shaking flask Liquid Culture, the culture medium that shaking flask Liquid Culture uses for PDA liquid medium,
PDA liquid medium includes that 200g potato and 20g glucose or sucrose make raw material, and potato is boiled 30min in water
20g glucose or sucrose is added in filtering in filtered supernatant, and water is added to be settled to 1000mL, and 121 DEG C of Yu Wendu~
125 DEG C, disinfecting action 0.5h is carried out under the conditions of the MPa of pressure 0.103MPa~0.168, then from above-mentioned PDA solid medium
Picking thallus is inoculated into PDA fluid nutrient medium, the shaken cultivation 24 under the conditions of revolving speed 200rpm~225rpm, 35 DEG C of temperature
~48h, obtains fermentation seed liquid, and the inoculum concentration is 2%.
7. the preparation method of the biological agent of prevention and treatment powdery mildew of strawberry according to claim 4, it is characterised in that: the step
Fermentation in rapid one uses fermentation tank culture, and fermentation seed liquid is gone in fermentor with 2% inoculum concentration and is fermented, first in temperature
121 DEG C~125 DEG C of degree, the potassium dihydrogen phosphate of pressure 0.103MPa~0.168 and 1000ml water, pH is 7.2~7.4, when gemma shape
When reaching 80% at rate, thallus fermentation process is completed, and stops fermentation, and the cell cracking in the step 1 is to by fermentor
Bacillus thallus after culture heats 15s~16s at 75 DEG C~90 DEG C of temperature, carries out high temperature, short time cracking, to be crushed bacterium
Body cell obtains zymocyte liquid.
8. under the conditions of MPa carry out sterilization treatment 1.0h, after ferment under the conditions of 35 DEG C of temperature, revolving speed 200rpm, ventilatory capacity 1:1,
The fermentation tank culture medium includes 10g peptone, 5g yeast extract, 3.5g glucose, 5g sodium chloride, 0.5g
The preparation method of the biological agent of prevention and treatment powdery mildew of strawberry according to claim 4, it is characterised in that: the step
Spray drying in three is dried using spray dryer, and it is 150 DEG C~160 DEG C that inlet air temperature is required in spray dryer,
Intake is 40000m3/ h~45000m3/ h, temperature of outgoing air are 70 DEG C~80 DEG C, exhaust air rate 40000m3/ h~45000m3/
H, mixing suspension is through high speed centrifugal atomization device, and rotating spraying is at imperceptible misty liquid droplets, and hot-air parallel flow contact, quickly
It is dried to powder, obtains dry primary biological agent.
9. the biological agent of prevention and treatment powdery mildew of strawberry according to claim 1 to 8, it is characterised in that: described anti-
Control the biological agent of powdery mildew of strawberry prevention and treatment powdery mildew of strawberry, uncinula necator, powdery mildew of cucumber, watermelon powdery mildew disease or
The application of melon powdery mildew.
10. the biological agent of prevention and treatment powdery mildew of strawberry according to claim 1 to 8, it is characterised in that: described
Prevention and treatment powdery mildew of strawberry biological agent be diluted with water 600~1000 times, be sprayed on crop blade face or to crop carry out pouring root,
Soak the application of root or seed soaking.
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