CN109169713A - A kind of Bei Laisi bacillus ZSY-1 wettable powder and preparation method and application - Google Patents
A kind of Bei Laisi bacillus ZSY-1 wettable powder and preparation method and application Download PDFInfo
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- CN109169713A CN109169713A CN201811056222.XA CN201811056222A CN109169713A CN 109169713 A CN109169713 A CN 109169713A CN 201811056222 A CN201811056222 A CN 201811056222A CN 109169713 A CN109169713 A CN 109169713A
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- 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
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- 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
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- A01N25/14—Powders or granules wettable
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Abstract
The invention discloses a kind of Bei Laisi bacillus ZSY-1 wettable powder and preparation method and application, are made of the raw material of following weight percent: Bei Laisi bacillus ZSY-1 fermentation liquid 70%, dispersing agent 3-10%, wetting agent 2-4%, stabilizer 2%, surplus are carrier wheat bran；It is that carrier wheat bran is uniformly mixed by Bei Laisi bacillus ZSY-1 fermentation liquid, dispersing agent, wetting agent, stabilizer, surplus, after freeze-dried machine is dry, crushing in pulverizer makes its fineness be less than or equal to 44 μm, is sealed up to Bei Laisi bacillus ZSY-1 wettable powder.Compared with prior art, in tomato gray mould disease occurs for this bacillus ZSY-1 wettable powder of Lay of the invention early period or initial stage uses, and can play with good prevention and treatment.
Description
Technical field
The present invention relates to tomato gray mould and early epidemic disease prevention and control field, especially a kind of Bei Laisi bacillus ZSY-1 can
Wet powder and preparation method and application.
Background technique
Tomato gray mould and early epidemic disease as 2 kinds of worldwide diseases during tomato cultivation, each cultivation area every year all
There is generation, not only there is great influence to tomato yield and quality, but also fresh-keeping there is to tomato postharvest storage and also important shadow
Ring, thus in recent years numerous researchers it is adopted before and postharvest disease have conducted extensive research.Research direction relates generally to chemistry
Prevention and treatment, physical control, resistant variety breeding, biological control and drug resistance and pests occurrence rule etc., wherein with chemical prevention and resisting
Pharmacological property research is more, is secondly biological control, and obtain certain progress.Although recently as chemical pesticide residual, drug resistance
And the appearance of the problems such as phytotoxicity, biological pesticide become new development direction, by vast plant protection work person numerous studies, and find a variety of
Plant extracts and Biocontrol microorganism have good control efficiency to tomato gray mould and early epidemic disease, but these researchs at present are also all
Be concentrated mainly on desk research and bacterial strain or its active metabolite control efficiency research, and study discovery part bacterial strain or activity at
Divide significant with Field information difference on effect indoors.In addition, active bacterial strain or number of plant and formulation development and Field information
Studying quantity, there is also larger gaps, especially studied in terms of endophytic bacterium prevents and treats tomato gray mould and early epidemic disease also compared with
For weakness.In addition, it is more with tomato gray mould disease research in the 2 kinds of biological control of diseases researchs of tomato early epidemic and grey mold, and kind
Eggplant early epidemic disease infects the disease based on blade as a kind of, equally has a great influence to Development of Tomato, and with
The discovery of resistance problems brings difficulty to its chemical prevention, therefore related endophytic bacterium prevents and treats tomato in later period research
Grey mold and early epidemic disease and by production practices apply for the purpose of active bacterial strain Field Screening, preparation processing and field control effect
Fruit research also needs to further strengthen.
Summary of the invention
The invention aims to solve deficiency present in prior art problem, a kind of Bei Laisi bacillus is provided
ZSY-1 wettable powder and preparation method and application.
In order to achieve the above objectives, the present invention is implemented according to following technical scheme:
A kind of Bei Laisi bacillus ZSY-1 wettable powder, is made: Bei Laisi bud of the raw material of following weight percent
Spore bacillus ZSY-1 fermentation liquid 70%, dispersing agent 3-10%, wetting agent 2-4%, stabilizer 2%, surplus are carrier wheat bran；Wherein,
The Bei Laisi bacillus ZSY-1 is deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation
Number are as follows: CGMCC No.16230, the deposit date is on August 8th, 2018.
Further, the dispersing agent are as follows: the sodium carboxymethylcellulose or weight percent that weight percent is 6% are 8%
Sodium lignin sulfonate or weight percent be 3% dispersing agent MF or weight percent be 10% three sodium hexametaphosphates or
The calcium lignosulfonate that weight percent is 8%.
Further, the wetting agent are as follows: the infiltration that the Nekal BX or weight percent that weight percent is 4% are 2%
The Silok-2235 that the Silok-7110 or weight percent that agent T or weight percent are 2% are 2%.
Further, the stabilizer is sodium carboxymethylcellulose.
In addition, the present invention also provides a kind of preparation method of Bei Laisi bacillus ZSY-1 wettable powder, including it is following
Step:
1) 20 μ L Bei Laisi bacillus ZSY-1 of -80 DEG C of preservations are taken to be coated on fresh NA plate, and in 26 DEG C of constant temperature
Activation culture for 24 hours, the Bei Laisi bacillus ZSY-1 after being activated；
2) the Bei Laisi bacillus ZSY-1 after taking 1 ring to activate using oese, is seeded to fresh BPY without fermented liquid
In, in 160r/min, 28 DEG C of fermented and cultured 5d, microscopy spore content is sent out after its resistance gemma rate up to termination after 90% or more
Ferment obtains Bei Laisi bacillus ZSY-1 fermentation liquid；
3) by carrier wheat bran, so that its fineness is less than or equal to 44 μm through pulverizer crushing, be loaded on 500mL indigo plant lid reagent bottle
Afterwards in autoclave sterilization pot sterilizing 30min, it is placed in 50 DEG C of baking ovens and is dried for standby；
4) Bei Laisi bacillus ZSY-1 fermentation liquid 70%, dispersing agent 3-10%, wetting agent 2- are taken according to weight percent
4%, stabilizer 2%, surplus are carrier wheat bran, are then uniformly mixed, and after freeze-dried machine is dry, crushing in pulverizer makes
Its fineness is less than or equal to 44 μm, is sealed up to Bei Laisi bacillus ZSY-1 wettable powder.
Furthermore the present invention also provides a kind of application of Bei Laisi bacillus ZSY-1 wettable powder, the Bei Laisi buds
Spore bacillus ZSY-1 wettable powder sterile water is used to prevent and treat botrytis cinerea and early blight of tomato after diluting 300-500 times
Bacterium.
Compared with prior art, this bacillus ZSY-1 wettable powder of Lay of the invention prevents and treats tomato gray mould disease
Optimum diluting multiple is 500 times, and early blight of tomato evil prevention and treatment optimum diluting multiple is 300 times, anti-under optimum diluting multiple
Effect is respectively 87.37% and 79.01%, and best field medication extension rate is 300 times, can prevent and treat 2 kinds of disease (grey mold simultaneously
Germ and early epidemic germ), and generated without phytotoxicity.This bacillus ZSY-1 wettable powder of Lay of the invention is in graw mold of tomato
Early period occurs for evil or initial stage uses, and can play with good prevention and treatment.
Specific embodiment
To make the objectives, technical solutions, and advantages of the present invention more comprehensible, with reference to embodiments, to the present invention into
Row is further to be described in detail.Described herein the specific embodiments are only for explaining the present invention, is not used to limit invention.
Embodiment 1
A kind of Bei Laisi bacillus ZSY-1 wettable powder, is made: Bei Laisi bud of the raw material of following weight percent
Spore bacillus ZSY-1 fermentation liquid 70%, sodium carboxymethylcellulose 6%, Nekal BX 4%, stabilizer sodium carboxymethylcellulose 2%,
Surplus is carrier wheat bran；Wherein, the Bei Laisi bacillus ZSY-1 is deposited in China Committee for Culture Collection of Microorganisms
Common micro-organisms center, deposit number are as follows: CGMCC No.16230, the deposit date is on August 8th, 2018.
In addition, the present invention also provides a kind of preparation method of Bei Laisi bacillus ZSY-1 wettable powder, including it is following
Step:
1) 20 μ L Bei Laisi bacillus ZSY-1 of -80 DEG C of preservations are taken to be coated on fresh NA plate, and in 26 DEG C of constant temperature
Activation culture for 24 hours, the Bei Laisi bacillus ZSY-1 after being activated；
2) the Bei Laisi bacillus ZSY-1 after taking 1 ring to activate using oese, is seeded to fresh BPY without fermented liquid
In, in 160r/min, 28 DEG C of fermented and cultured 5d, microscopy spore content is sent out after its resistance gemma rate up to termination after 90% or more
Ferment obtains Bei Laisi bacillus ZSY-1 fermentation liquid；
3) by carrier wheat bran, so that its fineness is less than or equal to 44 μm through pulverizer crushing, be loaded on 500mL indigo plant lid reagent bottle
Afterwards in autoclave sterilization pot sterilizing 30min, it is placed in 50 DEG C of baking ovens and is dried for standby；
4) Bei Laisi bacillus ZSY-1 fermentation liquid 70% is taken according to weight percent, sodium carboxymethylcellulose 6%, draw
Opening powder BX 4%, stabilizer sodium carboxymethylcellulose 2%, surplus is carrier wheat bran, is then uniformly mixed, and freeze-dried machine is dry
After dry, crushing in pulverizer makes its fineness be less than or equal to 44 μm, is sealed wettable up to Bei Laisi bacillus ZSY-1
Property pulvis.
Embodiment 2
A kind of Bei Laisi bacillus ZSY-1 wettable powder, is made: Bei Laisi bud of the raw material of following weight percent
It is spore bacillus ZSY-1 fermentation liquid 70%, sodium lignin sulfonate 8%, penetrant t 2%, stabilizer sodium carboxymethylcellulose 2%, remaining
Amount is carrier wheat bran；Wherein, it is general to be deposited in China Committee for Culture Collection of Microorganisms by the Bei Laisi bacillus ZSY-1
Logical microorganism center, deposit number are as follows: CGMCC No.16230, the deposit date is on August 8th, 2018.
In addition, the present invention also provides a kind of preparation method of Bei Laisi bacillus ZSY-1 wettable powder, including it is following
Step:
1) 20 μ L Bei Laisi bacillus ZSY-1 of -80 DEG C of preservations are taken to be coated on fresh NA plate, and in 26 DEG C of constant temperature
Activation culture for 24 hours, the Bei Laisi bacillus ZSY-1 after being activated；
2) the Bei Laisi bacillus ZSY-1 after taking 1 ring to activate using oese, is seeded to fresh BPY without fermented liquid
In, in 160r/min, 28 DEG C of fermented and cultured 5d, microscopy spore content is sent out after its resistance gemma rate up to termination after 90% or more
Ferment obtains Bei Laisi bacillus ZSY-1 fermentation liquid；
3) by carrier wheat bran, so that its fineness is less than or equal to 44 μm through pulverizer crushing, be loaded on 500mL indigo plant lid reagent bottle
Afterwards in autoclave sterilization pot sterilizing 30min, it is placed in 50 DEG C of baking ovens and is dried for standby；
4) Bei Laisi bacillus ZSY-1 fermentation liquid 70%, sodium lignin sulfonate 8%, infiltration are taken according to weight percent
Agent T 2%, stabilizer sodium carboxymethylcellulose 2%, surplus are carrier wheat bran, are then uniformly mixed, and freeze-dried machine is dry
Afterwards, crushing in pulverizer makes its fineness be less than or equal to 44 μm, is sealed up to Bei Laisi bacillus ZSY-1 wettable
Pulvis.
Embodiment 3
A kind of Bei Laisi bacillus ZSY-1 wettable powder, is made: Bei Laisi bud of the raw material of following weight percent
It is spore bacillus ZSY-1 fermentation liquid 70%, dispersing agent MF 3%, Silok-71102%, stabilizer sodium carboxymethylcellulose 2%, remaining
Amount is carrier wheat bran；Wherein, it is general to be deposited in China Committee for Culture Collection of Microorganisms by the Bei Laisi bacillus ZSY-1
Logical microorganism center, deposit number are as follows: CGMCC No.16230, the deposit date is on August 8th, 2018.
In addition, the present invention also provides a kind of preparation method of Bei Laisi bacillus ZSY-1 wettable powder, including it is following
Step:
1) 20 μ L Bei Laisi bacillus ZSY-1 of -80 DEG C of preservations are taken to be coated on fresh NA plate, and in 26 DEG C of constant temperature
Activation culture for 24 hours, the Bei Laisi bacillus ZSY-1 after being activated；
2) the Bei Laisi bacillus ZSY-1 after taking 1 ring to activate using oese, is seeded to fresh BPY without fermented liquid
In, in 160r/min, 28 DEG C of fermented and cultured 5d, microscopy spore content is sent out after its resistance gemma rate up to termination after 90% or more
Ferment obtains Bei Laisi bacillus ZSY-1 fermentation liquid；
3) by carrier wheat bran, so that its fineness is less than or equal to 44 μm through pulverizer crushing, be loaded on 500mL indigo plant lid reagent bottle
Afterwards in autoclave sterilization pot sterilizing 30min, it is placed in 50 DEG C of baking ovens and is dried for standby；
4) Bei Laisi bacillus ZSY-1 fermentation liquid 70%, dispersing agent MF 3%, Silok- are taken according to weight percent
71102%, stabilizer sodium carboxymethylcellulose 2%, surplus are carrier wheat bran, are then uniformly mixed, and freeze-dried machine is dry
Afterwards, crushing in pulverizer makes its fineness be less than or equal to 44 μm, is sealed up to Bei Laisi bacillus ZSY-1 wettable
Pulvis.
Embodiment 4
A kind of Bei Laisi bacillus ZSY-1 wettable powder, is made: Bei Laisi bud of the raw material of following weight percent
Spore bacillus ZSY-1 fermentation liquid 70%, three sodium hexametaphosphates 10%, Silok-2235 2%, stabilizer sodium carboxymethylcellulose
2%, surplus is carrier wheat bran；Wherein, the Bei Laisi bacillus ZSY-1 is deposited in Chinese microorganism strain preservation management committee
Member's meeting common micro-organisms center, deposit number are as follows: CGMCC No.16230, the deposit date is on August 8th, 2018.
In addition, the present invention also provides a kind of preparation method of Bei Laisi bacillus ZSY-1 wettable powder, including it is following
Step:
1) 20 μ L Bei Laisi bacillus ZSY-1 of -80 DEG C of preservations are taken to be coated on fresh NA plate, and in 26 DEG C of constant temperature
Activation culture for 24 hours, the Bei Laisi bacillus ZSY-1 after being activated；
2) the Bei Laisi bacillus ZSY-1 after taking 1 ring to activate using oese, is seeded to fresh BPY without fermented liquid
In, in 160r/min, 28 DEG C of fermented and cultured 5d, microscopy spore content is sent out after its resistance gemma rate up to termination after 90% or more
Ferment obtains Bei Laisi bacillus ZSY-1 fermentation liquid；
3) by carrier wheat bran, so that its fineness is less than or equal to 44 μm through pulverizer crushing, be loaded on 500mL indigo plant lid reagent bottle
Afterwards in autoclave sterilization pot sterilizing 30min, it is placed in 50 DEG C of baking ovens and is dried for standby；
4) according to weight percent take Bei Laisi bacillus ZSY-1 fermentation liquid 70%, three sodium hexametaphosphates 10%,
Silok-2235 2%, stabilizer sodium carboxymethylcellulose 2%, surplus are carrier wheat bran, are then uniformly mixed, freeze-dried
After machine is dry, crushing in pulverizer makes its fineness be less than or equal to 44 μm, is sealed up to Bei Laisi bacillus ZSY-1
Wettable powder.
Embodiment 5
A kind of Bei Laisi bacillus ZSY-1 wettable powder, is made: Bei Laisi bud of the raw material of following weight percent
It is spore bacillus ZSY-1 fermentation liquid 70%, calcium lignosulfonate 8%, Nekal BX 4%, stabilizer sodium carboxymethylcellulose 2%, remaining
Amount is carrier wheat bran；Wherein, it is general to be deposited in China Committee for Culture Collection of Microorganisms by the Bei Laisi bacillus ZSY-1
Logical microorganism center, deposit number are as follows: CGMCC No.16230, the deposit date is on August 8th, 2018.
In addition, the present invention also provides a kind of preparation method of Bei Laisi bacillus ZSY-1 wettable powder, including it is following
Step:
1) 20 μ L Bei Laisi bacillus ZSY-1 of -80 DEG C of preservations are taken to be coated on fresh NA plate, and in 26 DEG C of constant temperature
Activation culture for 24 hours, the Bei Laisi bacillus ZSY-1 after being activated；
2) the Bei Laisi bacillus ZSY-1 after taking 1 ring to activate using oese, is seeded to fresh BPY without fermented liquid
In, in 160r/min, 28 DEG C of fermented and cultured 5d, microscopy spore content is sent out after its resistance gemma rate up to termination after 90% or more
Ferment obtains Bei Laisi bacillus ZSY-1 fermentation liquid；
3) by carrier wheat bran, so that its fineness is less than or equal to 44 μm through pulverizer crushing, be loaded on 500mL indigo plant lid reagent bottle
Afterwards in autoclave sterilization pot sterilizing 30min, it is placed in 50 DEG C of baking ovens and is dried for standby；
4) Bei Laisi bacillus ZSY-1 fermentation liquid 70% is taken according to weight percent, calcium lignosulfonate 8%, pull open
Powder BX 4%, stabilizer sodium carboxymethylcellulose 2%, surplus are carrier wheat bran, are then uniformly mixed, and freeze-dried machine is dry
Afterwards, crushing in pulverizer makes its fineness be less than or equal to 44 μm, is sealed up to Bei Laisi bacillus ZSY-1 wettable
Pulvis.
Embodiment 6
A kind of Bei Laisi bacillus ZSY-1 wettable powder, is made: Bei Laisi bud of the raw material of following weight percent
Spore bacillus ZSY-1 fermentation liquid 70%, sodium carboxymethylcellulose 6%, penetrant t 2%, stabilizer sodium carboxymethylcellulose 2%,
Surplus is carrier wheat bran；Wherein, the Bei Laisi bacillus ZSY-1 is deposited in China Committee for Culture Collection of Microorganisms
Common micro-organisms center, deposit number are as follows: CGMCC No.16230, the deposit date is on August 8th, 2018.
In addition, the present invention also provides a kind of preparation method of Bei Laisi bacillus ZSY-1 wettable powder, including it is following
Step:
1) 20 μ L Bei Laisi bacillus ZSY-1 of -80 DEG C of preservations are taken to be coated on fresh NA plate, and in 26 DEG C of constant temperature
Activation culture for 24 hours, the Bei Laisi bacillus ZSY-1 after being activated；
2) the Bei Laisi bacillus ZSY-1 after taking 1 ring to activate using oese, is seeded to fresh BPY without fermented liquid
In, in 160r/min, 28 DEG C of fermented and cultured 5d, microscopy spore content is sent out after its resistance gemma rate up to termination after 90% or more
Ferment obtains Bei Laisi bacillus ZSY-1 fermentation liquid；
3) by carrier wheat bran, so that its fineness is less than or equal to 44 μm through pulverizer crushing, be loaded on 500mL indigo plant lid reagent bottle
Afterwards in autoclave sterilization pot sterilizing 30min, it is placed in 50 DEG C of baking ovens and is dried for standby；
4) Bei Laisi bacillus ZSY-1 fermentation liquid 70% is taken according to weight percent, sodium carboxymethylcellulose 6%, seep
Saturating agent T 2%, stabilizer sodium carboxymethylcellulose 2%, surplus are carrier wheat bran, are then uniformly mixed, freeze-dried machine is dry
After dry, crushing in pulverizer makes its fineness be less than or equal to 44 μm, is sealed wettable up to Bei Laisi bacillus ZSY-1
Property pulvis.
Further, Bei Laisi bacillus ZSY-1 wettable powder storage-stable is measured:
Bei Laisi bacillus ZSY-1 wettable powder is dispensed, every bag of net weight 10g, and protected in a cool and dry place
It deposits.
Storage-stable measurement
1. light durability
Bei Laisi bacillus ZSY-1 wettable powder prevents and treats biological agent as a kind of tomato field diseases, in natural light
Ultraviolet light its activity may be generated it is certain influence, therefore this test is in obtained Bei Laisi bacillus ZSY-1 wettable
After pulvis, its stability is measured under natural lighting again.
After obtained Bei Laisi bacillus ZSY-1 wettable powder on March 8th, 2016, on March 15th, 2016 extremely
On April 14th, 2016, daily from 9 illumination to 4 points, continuous illumination 30d, during which every 5d to its viable count and bacteriostatic activity into
Row detection, so that it is determined that the light durability of Bei Laisi bacillus ZSY-1 wettable powder.
2. room temperature storage stability
Bei Laisi bacillus ZSY-1 wettable powder is prepared, and in room temperature storage after packing, during which every January to it
Viable count and bacteriostatic activity are detected, continuous detection 12 months, to determine Bei Laisi bacillus ZSY-1 wettable powder
Storage-stable.
It is found after the light durability to Bei Laisi bacillus ZSY-1 wettable powder detects, this is wettable
Property pulvis there is good light durability, viable count declines unobvious (referring to table 1) after outdoor placement 30d in valve bag.
It finds that the decline of its bacteriostatic activity is also unobvious after measuring bacteriostatic activity using tablet face-off method, illustrates Bei Laisi bacillus ZSY-
1 wettable powder has preferable light durability, and storing property is good, lays a good foundation for the holding of its field efficacy.
Influence of the different light application times of table 1 to Bei Laisi bacillus ZSY-1 wettable powder stability
It is sent out after being measured using storage-stable of the room temperature storage to Bei Laisi bacillus ZSY-1 wettable powder
Existing, number of viable decline is unobvious after bacterial strain ZSY-1 wettable powder room temperature storage 1 year, only has dropped compared with storage 1 month
4.85%, illustrate that bacterial strain ZSY-1 wettable powder has preferable storage-stable；In terms of bacteriostatic activity, plate pair is used
It stands erect after method is detected using tomato early blight bacterium as target and finds, bacteriostatic activity decline is unobvious, bacteriostatic activity after storage 1 year
Remain at 86.19% (referring to table 2).In addition, though wettable powder bacteriostatic activity extends bacteriostatic activity with storage time
There is slightly decline, but fall is the same as, without linear dependence, this non-linear relation may be with antibacterial work between storage time
Property detection when test error it is related, illustrate that bacterial strain ZSY-1 can still keep preferable bacteriostatic activity after room temperature storage 1 year, be its
Later period commercialization is laid a good foundation.
2 Bei Laisi bacillus ZSY-1 wettable powder room temperature storage stability of table
Further, Bei Laisi bacillus ZSY-1 wettable powder to the tomato of B.cinerea and A. solani from
Body control effect testing:
It chooses tamato fruit similar in size and maturity and uses wound inocalation method, measure Bei Laisi bacillus ZSY-1
To the living body fungistatic effect of 2 kinds of cause of diseases of B.cinerea and A.solani, 3 fruits of every processing, each test is repeated 3 times.Stream
Journey is as follows: (1) surface sterilization: 75% ethyl alcohol impregnate 5min, 3% liquor natrii hypochloritis impregnate 5min, aseptic water washing 3 times, nothing
Bacterium filter paper is used for follow-up test after drying；(2) prepared by wound: diameter being used gently to punch for the punch of 5mm on tomato surface
Diameter is the hole of 5mm, and prevents skin crack；(3) Bei Laisi bacillus ZSY-1 wettable powder is inoculated with: wettable powder
Agent draws 150 μ L bacteria suspensions after sterile water dilutes 10 times, using pipettor, is seeded to fruit wound, is pair with sterile water
According to using transfer needle picking diameter for the disease fungus fungus block of 5mm, and be seeded to fruit after liquid is fully absorbed by fruit
Wound；(4) it cultivates: after inoculation, each fruit being placed in sterile porcelain dish, and be put into 2 pieces in blank space and speckle with the de- of sterile water
Rouge cotton is used for moisturizing.It is finally placed on using preservative film sealing after cultivating 7d in 26 DEG C of constant incubators and measures lesion area, and
Calculate preventive effect.
Lesion area (cm2)=π R2(R: fruit surface scab radius cm)；
Preventive effect %=(control lesion area-processing lesion area) × 100/ control lesion area.
Using botrytis cinerea and tomato early blight bacterium as target, using punching inocalation method to Bei Laisi bacillus ZSY-
1 wettable powder is found after being measured on tamato fruit to the fungistatic effect of 2 kinds of disease fungus: Bei Laisi bacillus
ZSY-1 wettable powder all has preferable in vitro preventive effect to botrytis cinerea and tomato early blight bacterium, and bacteriostasis rate is respectively
91.42% and 86.34% (table 3).In addition, tamato fruit, which is inoculated with 2 kinds of cause of disease back edges, there is water stain shape putrefactive phenomenon,
In it is the most obvious with inoculating tomato grey mold cause of disease, illustrate 2 kinds except can be in addition to tomato is endangered in field or during tomato postharvest storage
2 kinds of pathogens.
3 Bei Laisi bacillus ZSY-1 wettable powder of table prevents botrytis cinerea and the in vitro of tomato early blight bacterium
Effect
Further, early to tomato tomato gray mould disease and tomato for determining Bei Laisi bacillus ZSY-1 wettable powder
The control efficiency and optimum diluting multiple of epidemic disease evil, choose size and the close tamato fruit of maturity, right using punching inocalation method
Bei Laisi bacillus ZSY-1 wettable powder prevents and treats effect to the tamato fruit of 2 kinds of tomato diseases in vitro under different extension rates
Fruit is measured, the specific steps are as follows: 1. surface sterilization: tamato fruit impregnates 5min through 3%NaClO and 75% ethyl alcohol respectively,
Sterile water wash 3 times, aseptic filter paper is used for follow-up test after drying surface；2. punching: the punch for the use of diameter being 5mm,
The aperture for taking diameter to be 5mm is beaten on tomato surface, is used for inoculating tomato ash arrhizus bacteria, tomato early blight bacterium and Bei Laisi gemma bar
Bacterium ZSY-1 wettable powder；3. wettable powder dilution agent: wettable powder is diluted respectively using sterile water 0 times, 100 times,
300 times, 500 times and 1000 times, using equivalent without Bei Laisi bacillus ZSY-1 blank formulation as compare；4. inoculation: point
It does not draw the 120 μ L of wettable powder dilution under different extension rates to be added in the wound that diameter is 5mm, to its liquid
After infiltration completely, using transfer needle by diameter is the botrytis cinerea of 5mm and tomato early blight bacterium pure culture biscuits involvng inoculation into wound；
5. culture: finishing and be placed in sterile tray in inoculation, be put into the sterile absorbent progress moisturizing for speckling with sterile water, and use preservative film
It is closed at 25 DEG C after being protected from light culture 7d, calculates the diseases prevention of Bei Laisi bacillus ZSY-1 wettable powder difference extension rate
Effect.
Using tomato isolated test to Bei Laisi bacillus ZSY-1 wettable powder in early blight of tomato evil and tomato ash
Extension rate during mould disease control is found after being measured, and under different extension rates, Bei Laisi bacillus ZSY-1 can
Wet powder is to 2 kinds of disease-controlling effect significant differences.It is 100 times in extension rate, bacterial strain ZSY-1 wettable powder preventive effect is same
Stoste is not significant compared to difference；Although being that the control efficiency of bacterial strain ZSY-1 wettable powder goes out when extension rate increases to 300 times
Now decline to a certain degree, and the significant difference compared with 100 times and 0 times, but still preferable control efficiency can be maintained, to early blight of tomato
The preventive effect of bacterium and botrytis cinerea is respectively 79.01% and 88.44%；When extension rate is 500 times, bacterial strain ZSY-1 is wettable
Property pulvis larger difference is then shown to the control efficiency of 2 kinds of diseases, at this time to early blight of tomato evil control efficiency decline compared with
To be obvious, control efficiency is reduced to 76.31%, and then unobvious to the decline of the control efficiency of tomato gray mould disease, is also positively retained at
87.37%, illustrate that the bacterial strain ZSY-1 extension rate required when preventing and treating 2 kinds of diseases is different, in early blight of tomato evil prevention and treatment
It is advisable for 300 times with dilution, and is advisable in the prevention and treatment of tomato gray mould disease with diluting 500 times；When extension rate reaches 1000 times,
Occur largely declining to the control efficiency of 2 kinds of diseases, preventive effect is below 50%.Therefore, in prevention and treatment early blight of tomato evil
When select dilution 300 times as optimum diluting multiple；500 times of extension rates are selected when preventing and treating tomato gray mould disease；And same
When 2 kinds of diseases of prevention and treatment when then select 300 times of extension rates, so the dilution of Bei Laisi bacillus ZSY-1 wettable powder times
Number is 300~500 times (table 4).
The different extension rate Bei Laisi bacillus ZSY-1 wettable powders of table 4 prevent tomato early epidemic and grey mold disease
Control effect
In turn, the drug effect of Bei Laisi bacillus ZSY-1 wettable powder Field information is measured, specific as follows:
Reagent agent
Treatment agent: Bei Laisi bacillus ZSY-1 wettable powder (active constituent content: 400,000,000 living spores/gram)
Comparison medicament: 50% procymidone wettable powder (Shandong Xin Xing pesticide Co., Ltd)
For studying object
Tomato in plastic greenhouse, kind are as follows: powder general, nova 101, hat difficult to understand No. 6 and hundred million hard powder are won in agriculture
Experimental design
Processing design: 300 times of Bei Laisi bacillus wettable powder (A) and 500 times of dilutions (B)；Comparison medicament
600 times of dilutions (C) of 50% procymidone and total 4 processing of clear water blank control (D).
Quarter Design: small Division, every cell 15m2, every place are carried out to test greenhouse using single factor test RANDOMIZED BLOCK DESIGN
Manage 4 repetitions (table 5).
5 graw mold of tomato field efficacy RANDOMIZED BLOCK DESIGN of table
| District's groups 1 | C | B | C | C |
| District's groups 2 | A | D | B | A |
| District's groups 3 | D | C | D | D |
| District's groups 4 | B | A | A | B |
Experimental condition and application method
It is experimental field respectively the village Xiangfen County Chen Guo tomato species growing area, the Xinjiang County village great Nie tomato species growing area, Taigu County Yang Yi
Tomato species growing area and Xinjiang County north Zhang Zhen head west village village tomato species growing area, and each experimental field nearly 3 have graw mold of tomato every year.
Experimental field administration time is respectively on April 8th, 2016, on July 6th, 2016, on October 12nd, 2016 and in May, 2017 for the first time
9, the medication interval time was 7d, and continuous use 3 times, tomato is carried out by normal management during test.
Investigation content and method
(1) safety observations
2 kinds of Bei Laisi bacillus ZSY-1 wettable powder processing are observed during medication, and to tomato growth, whether there is or not phytotoxicity to show
As.
(2) preventive effect is investigated
This test carries out 2 disease survey altogether, is prevented after the investigation of state of an illness radix and last application 10d before being respectively administered
Effect investigation.Each cell is all made of 5 point samplings and is sampled, and every randomly selects 3 plants, investigates whole blades and the fruit state of an illness refers to
Number, calculates preventive effect, and using the Deng Kenshi duncan's new multiple range method in SPSS19.0 software under the level of P < 0.05 to test data into
Row statistical analysis.
Graw mold of tomato grade scale (as unit of blade) are as follows:
0 grade: disease-free spot；
1 grade: single blade has scab 3；
3 grades: single blade has scab 4~6；
5 grades: single blade has scab 7~10；
7 grades: single blade has scab 11~20, and part is intensive in blocks；
9 grades: single blade has scab intensively to account for blade area a quarter or more.
Fruit gray mold stage division:
0 grade: disease-free spot；
1 grade: the morbidity of residual petal or column cap morbidity；
3 grades: sepal rots or column cap morbidity spreads to areola portion；
5 grades: areola portion has infiltration spot without mould layer；
7 grades: areola portion has infiltration spot to have mould layer but unexpanded to other positions；
9 grades: areola portion has infiltration spot to have mould layer and have been expanded to other positions.
(3) preventive effect calculation method
Disease index=[(Σ (disease leaf (fruit) numbers at different levels × opposite value of series)/(investigating total leaf (fruit) number × 9)] × 100
Control efficiency (%)=(1-CK1 × T1/ (CK2 × T0)) × 100
In formula: CK1, CK2 are respectively that blank district is administered forward and backward disease index；T0, T1 are respectively chemicals treatment
Area is administered forward and backward disease index.
It is measured by the drug effect to Bei Laisi bacillus ZSY-1 wettable powder Field information, obtains measurement knot
Fruit are as follows:
1) tomato crop safety
Field efficacy is carried out in 300 times and 500 times dilutions of Bei Laisi bacillus ZSY-1 wettable powder during test
While test, follow-up observation also is carried out to its tomato crop safety, medicament pacifies crop after field spray use
Entirely, each trial zone do not occur suppressed tomato growth, yellow leaf, leaf rolling and plant wither here etc. bad phenomenons, explanation
Bacterial strain ZSY-1 wettable powder is to tomato crop safety.
2) tomato gray mould disease field efficacy
It is with 50% procymidone wettable powder of chemical agent in 4 different tomato species growing areas using RANDOMIZED BLOCK DESIGN
Comparison medicament, using clear water as blank control, 300 times of dilutions to Bei Laisi bacillus ZSY-1 wettable powder and 500 times
Dilution finds that the dilution of bacterial strain ZSY-1 wettable powder difference is again after being measured to the field control effect of graw mold of tomato
There is some difference for preventive effect of the number between different growing areas, and the control efficiency of 300 times of dilution is better than 500 times, and in great Nie
Village's growing area and the control efficiency to head west on the growing area blade of the village are generally better than fruit, and then show in old Guo and Yang city growing area
It is better than blade control efficiency for fruit control efficiency；In terms of blade preventive effect, same to comparison medicament (50% procymidone wettable powder
Agent) it compares, each 300 dilution blade control efficiency of tomato species growing area Bei Laisi bacillus ZSY-1 wettable powder is the same as control
Medicament is compared to there is some difference, except the ZSY-1300 times of dilution preventive effect of village area Bei Laisi bacillus that head west
(77.76%) same to comparison medicament (80.22%) is outside, and the preventive effect of remaining tomato species growing area bacterial strain ZSY-1 wettable powder is then
Significantly lower than comparison medicament；In terms of fruit field efficacy, 300 times of dilutions of bacterial strain ZSY-1 wettable powder are removed in Yang Yi kind
Outside growing area (61.36%) same to comparison medicament (70.73%) significant difference, remaining each growing area preventive effect is close poor with comparison medicament
It is different not significant.But when extension rate is 500, control efficiency is then significantly lower than comparison medicament, and significant difference；In addition, to phase
With bacterial strain ZSY-1 wettable powder under extension rate blade preventive effect and fruit preventive effect compare after find, except the village kind that heads west
Outside growing area, remaining each growing area shows as fruit preventive effect better than blade preventive effect.Illustrate Bei Laisi bacillus ZSY-1 wettable
The best field medication extension rate of pulvis is 300 times.
It finds after analyzing disease index before medication, is averaged disease index highest with positive city's medicine front vane and fruit,
Respectively 1.58 and 7.15, it is secondly the village great Nie, respectively 0.96 and 5.89, and it is minimum with average index before the village medicine that heads west, point
It Wei 0.75 and 4.66.Disease index finds that the low regional medicament of disease index is anti-after carrying out Conjoint Analysis with effect of chemical control
Effect is universal preferable, illustrate Bei Laisi bacillus ZSY-1 wettable powder at the initial stage of a disease medication when control efficiency it is preferable, have
Preferable prevention effect, referring to table 6.
Field efficacy of the 6 Bei Laisi bacillus ZSY-1 wettable powder of table to graw mold of tomato
Summary, this bacillus ZSY-1 wettable powder of Lay are 500 to tomato gray mould disease prevention and treatment optimum diluting multiple
Times, early blight of tomato evil prevention and treatment optimum diluting multiple is 300 times, and the preventive effect under optimum diluting multiple is respectively 87.37% He
79.01%, best field medication extension rate is 300 times, can prevent and treat 2 kinds of diseases (ash arrhizus bacteria and early epidemic germ) simultaneously,
And it is generated without phytotoxicity.The lower area of disease index before medicine, for blade field efficacy up to 77.76%, fruit drug effect is reachable
74.68%, it is suitable with 50% procymidone drug effect of comparison medicament (80.22% and 76.55%)；Disease index is significantly before medicine
Declining to a certain degree then occur in area, preventive effect, and drug effect is only 60% or so.Illustrate, wettable powder made from the bacterial strain is in tomato
Early period occurs for grey mold disease or initial stage uses, and can play with good prevention and treatment.
The limitation that technical solution of the present invention is not limited to the above specific embodiments, it is all to do according to the technique and scheme of the present invention
Technology deformation out, falls within the scope of protection of the present invention.
Claims (6)
1. a kind of Bei Laisi bacillus ZSY-1 wettable powder, which is characterized in that by the raw material system of following weight percent
At: Bei Laisi bacillus ZSY-1 fermentation liquid 70%, dispersing agent 3-10%, wetting agent 2-4%, stabilizer 2%, surplus are to carry
Body wheat bran；Wherein, the Bei Laisi bacillus ZSY-1 is deposited in China Committee for Culture Collection of Microorganisms's commonly micro- life
Object center, deposit number are as follows: CGMCC No.16230, the deposit date is on August 8th, 2018.
2. Bei Laisi bacillus ZSY-1 wettable powder according to claim 1, it is characterised in that: the dispersing agent
Are as follows: the sodium lignin sulfonate or weight percent that the sodium carboxymethylcellulose or weight percent that weight percent is 6% are 8%
The lignin sulfonic acid that three sodium hexametaphosphates or weight percent that dispersing agent MF or weight percent for 3% are 10% are 8%
Calcium.
3. Bei Laisi bacillus ZSY-1 wettable powder according to claim 1, it is characterised in that: the wetting agent
Are as follows: the penetrant t or weight percent that the Nekal BX or weight percent that weight percent is 4% are 2% are 2%
The Silok-2235 that Silok-7110 or weight percent are 2%.
4. Bei Laisi bacillus ZSY-1 wettable powder according to claim 1, it is characterised in that: the stabilizer
For sodium carboxymethylcellulose.
5. a kind of preparation method of Bei Laisi bacillus ZSY-1 wettable powder as described in claim 1, feature exist
In, comprising the following steps:
1) 20 μ L Bei Laisi bacillus ZSY-1 of -80 DEG C of preservations are taken to be coated on fresh NA plate, and in 26 DEG C of constant temperature activation trainings
It supports for 24 hours, the Bei Laisi bacillus ZSY-1 after being activated；
2) the Bei Laisi bacillus ZSY-1 after taking 1 ring to activate using oese, is seeded in fresh BPY without fermented liquid, in
160r/min, 28 DEG C of fermented and cultured 5d, microscopy spore content are obtained after its resistance gemma rate up to termination fermentation after 90% or more
To Bei Laisi bacillus ZSY-1 fermentation liquid；
3) by carrier wheat bran, so that its fineness is less than or equal to 44 μm through pulverizer crushing, loaded on after 500mL indigo plant lid reagent bottle in
High temperature and pressure (121 DEG C, 20min) autoclave sterilizing 30min, is placed in 50 DEG C of baking ovens and is dried for standby；
4) according to weight percent take Bei Laisi bacillus ZSY-1 fermentation liquid 70%, dispersing agent 3-10%, wetting agent 2-4%,
Stabilizer 2%, surplus are carrier wheat bran, are then uniformly mixed, and after freeze-dried machine is dry, crushing in pulverizer keeps it thin
Degree is less than or equal to 44 μm, is sealed up to Bei Laisi bacillus ZSY-1 wettable powder.
6. a kind of application of Bei Laisi bacillus ZSY-1 wettable powder as described in claim 1, it is characterised in that: institute
State Bei Laisi bacillus ZSY-1 wettable powder sterile water dilute 300-500 times after for prevent and treat botrytis cinerea with
Tomato early blight bacterium.
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