CN106967754A - A kind of technology that insecticide is prepared by fermenting raw materials of kuh-seng - Google Patents
A kind of technology that insecticide is prepared by fermenting raw materials of kuh-seng Download PDFInfo
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- CN106967754A CN106967754A CN201710243507.3A CN201710243507A CN106967754A CN 106967754 A CN106967754 A CN 106967754A CN 201710243507 A CN201710243507 A CN 201710243507A CN 106967754 A CN106967754 A CN 106967754A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
- C12P1/04—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using bacteria
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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
- 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
- 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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- 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
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
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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
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
- A01N65/08—Magnoliopsida [dicotyledons]
- A01N65/20—Fabaceae or Leguminosae [Pea or Legume family], e.g. pea, lentil, soybean, clover, acacia, honey locust, derris or millettia
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- 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
- C12N1/205—Bacterial isolates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
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- General Engineering & Computer Science (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Pest Control & Pesticides (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
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- Tropical Medicine & Parasitology (AREA)
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Abstract
The invention provides a kind of method that kuh-seng that ferments prepares insecticide, its process characteristic is using kuh-seng as primary raw material, it is aided with house refuse, starchy material and water, using commercially available bacillus pumilus, Su Yun bacillus and bacillus amyloliquefaciens H47 as fermentation strain, according to kuh-seng：House refuse：Starchy material:Pure water=2~5:1~2:0.01:3~7 ratio and the inoculation amount of system 2~4%, fermentation temperature are 25~35 DEG C, and rotating speed is 100~200rpm, keeps pH=5~8,6~9d of fermentation time.Complete fermentation of the present invention, makes a variety of enzymes therein, vitamin, amino acid and trace element fully discharge, product is improved the drug effect and its function of insecticide.And do not influence the physiological activator of botanical pesticide again.
Description
Technical field
A kind of method that microbial fermentation prepares insecticide, is related to and is conciliate with commercially available bacillus pumilus, Su Yun bacillus
Bacillus amyloliquefacienses H47 freeze-dried powders, the method that insecticide is prepared by fermenting raw materials of kuh-seng belongs to bioengineering and fermentation technique
Field.
Background technology
Kuh-seng (Sophora flavescens) is pulse family kuh-seng platymiscium, fall leaves fruticuli, high 1.5-3m.Root cylinder
Shape, crust yellow-white.Stem is upright, multi-branched, has longitudinal furrow；Sprout is become hairless afterwards by thin hair.Imparipinnate leaf, long 20-
25cm, alternate；Leaflet 15-29, blade lanceolar is to wire lanceolar, and long 3-4cm, wide 1.2-2cm, tip is tapering, base portion circle,
There are short handle, full edge, the close life patch pubescence in the back side；Stipule is linear.Raceme basidixed, long 15-20cm, by undercoat, bract is linear；
Calyx mitriform, flat, long 6-7mm, 5 shallow split；Corolla butterfly, yellowish white；Vexil cochlear, ala earless is isometric with fossil fragments；It is male
Stamen 10, filigree separation；Ovary handle is by fine, soft fur, and column cap is circular.Pod is linear, and tip has long beak, is not ftractureed when ripe, long 5-8cm.
Micro- contracting of hanging between seed, in unconspicuous beading, sparsely grow pubescence.3-7, seed, subsphaeroidal, black.The month at florescence 6-7, fruiting period
The 7-9 months.All parts of the country are distributed in, are born in sand ground or endroit thick grass and small stream limes marginis.Kuh-seng contains many secondary metabolism productions
Thing, kuh-seng ether extract and alcohol extract have stronger bacteriostasis to staphylococcus aureus；Kuh-seng flooding agent to trichophyton,
Trichophyton concentricum, Trichophyton schoenleini, Du small gemma tinea bacterium of big belly difficult to understand etc. have inhibitory action.Matrine can benumb nerve center, make
Polypide protein coagulating, closes polypide stomata, insect is choked to death.To the cabbage caterpillar on various crops, aphid, red spider etc.
Insect has obvious prevention effect.But botanical pesticide use organic solvent or decoct extract, 1) be unfavorable for energy-conservation also have compared with
Big environmental risk；2) stability of influence active material and loss active material；3) increase cost causes development and application difficult
Larger, action effect is slow, therefore brings huge financial difficulties to the research and development and production of plant insecticide.Using suitable
Plant growth-promoting bacteria fermentation kuh-seng, not only contribute to the extraction of physiological activator, be also less prone to cause the inactivation of active material, promote
Raw bacterium also has preferable disinsection prophylaxis and growth-promoting functions in itself.Be conducive to improving plant growth and strengthen insecticidal effect.Be conducive to
The effective utilization of environmental protection and kuh-seng plant in itself.
The content of the invention
The present invention is carried out fermentation by microorganism and is produced Antioxidative Factors and plant growth-promoting bacteria using the affluent resources of kuh-seng
Producing the multiclass such as enzyme, alcohols and aromatic substance has adopted metabolite to improve the drug effect and its function of insecticide, the present invention
Possess fermentation time short, harmful microorganism grow it is few even without, it is to avoid due to the virus crossed lengthy fermentation and produced, with
And the advantages of the physiological activators such as lipopeptid can be produced, so as to improve effect of disinsection prophylaxis and the growth-promoting functions of plant growth.
Inventive formulation and usage：
(1) selected material：It is fermentation main material to select the measured kuh-seng of fresh, matter.
(2) processing of material：First rinsed with running water, a century of Wuxi a century Shang Shui drinks Co., Ltd offer is provided
Still water clear water is washed twice, is finally invaded and steeped 120 seconds with 0.9% NaCl solution again, drained and standby.
(3) Spawn incubation：The century-old still water provided using Wuxi a century Shang Shui drinks Co., Ltd prepares LB culture mediums, will
Original preserves bacterial strain activation, cultivates the bacterium solution obtained for about 48 hours standby；
(4) inoculation fermentation：According to inoculative proportion 2~4%, commercially available bacillus pumilus, Su Yun bacillus are conciliate into starch
Bacillus H47 accesses fermenation raw liquid according to a certain percentage, and each bacteria concentration is 106CFU/ml, ferment 3~9d, pH value 5.0~
8.0,25 DEG C~35 DEG C of temperature.
(5) post-processing makes：Coarse filtration, tentatively screens out bulky grain residue, it is to avoid kuh-seng, which is put to easily cause long, to become sour, mistake
Filter condition：Sieves number is 20~32 mesh.Thin filtering：Small particle residue is further filtered, liquid is presented limpider
State, filter condition：Sieves number is 80~100 mesh, has both been able to the insecticide of kuh-seng fermentation raw material.
The preparation method of step (1) raw material comprises the following steps：
(11) instrument, utensil used in are intended to sterilizing, without greasy.
(12) kuh-seng is cleaned with pure water, then drained with the saline sook of pure water or 0.9%.
(13) kuh-seng is placed in sterilized container again, it is rear to add house refuse and starchy material and pure water, its
Volume ratio kuh-seng in middle fermenation raw liquid：House refuse：Starchy material:Pure water=2~5:1~2:0.01:3~7, produce hardship
Join fermenation raw liquid.
The beneficial effect of invention：
This is not only contributed to the extraction of physiological activator, is kept activity using suitable plant growth-promoting bacteria fermentation kuh-seng
The stability of material, Promoting bacteria can be inhibited in itself by dominant strain other miscellaneous bacterias grow so as to have preferable disinsection prophylaxis and
Growth-promoting functions.Be conducive to improving plant growth and strengthen insecticidal effect.Be conducive to environmental protection and kuh-seng plant in itself effective
Utilize.Improve the economic value of insecticide.
Embodiment
Below in conjunction with the embodiment of the present invention, technical scheme is carried out to describe clear, perfectly, it is clear that institute
The embodiment of description is only a part of embodiment of the invention, rather than whole embodiments.Based on the embodiment in the present invention,
The other embodiment that those of ordinary skill in the art are obtained on the premise of creative work is not made, belongs to the present invention
The scope of protection.
Embodiment 1
A kind of method that insecticide is prepared by fermenting raw materials of kuh-seng, comprises the following steps：
(1) the selection measured kuh-seng of fresh, matter is as fermentation primary raw material.
(2) pure water clear water is washed twice, is finally soaked, then cold sterilization drains, filled with 0.9% NaCl solution again
Enter in the cleaned container of pure water, probably account for the 20% of volume, add the house refuse that accounts for total liquid volume 5% and 1%
Starchy material is that fermenation raw liquid is standby.
(3) LB culture mediums are prepared, original is preserved into bacterial strain activation, the zymophyte that 80r/min shaken cultivations are obtained for about 48 hours
Liquid is standby；
(4) commercially available bacillus pumilus, Su Yun bacillus and bacillus amyloliquefaciens H47 are pressed according to inoculative proportion 3%
Deng than taking access fermenation raw liquid, 100r/min shaken cultivations fermentation 6d, pH value 7.0,25 DEG C of temperature respectively.
(5) coarse filtration is first carried out, sieves number is 20~32 mesh.Then thin filtering, sieves number is 80~100
Mesh, produces the insecticide that kuh-seng is fermentation raw material.
Embodiment 2
A kind of method that insecticide is prepared by fermenting raw materials of kuh-seng, comprises the following steps：
(1) the selection measured kuh-seng of fresh, matter is as fermentation primary raw material.
(2) pure water clear water is washed twice, is finally soaked, then cold sterilization drains, filled with 0.9% NaCl solution again
Enter in the cleaned container of pure water, probably account for the 30% of volume, add the house refuse that accounts for total liquid volume 10% and 1%
Starchy material is that fermenation raw liquid is standby.
(3) LB culture mediums are prepared, original is preserved into bacterial strain activation, the zymophyte that 100r/min shaken cultivations are obtained for about 48 hours
Liquid is standby；
(4) according to different vaccination ratio (commercially available bacillus pumilus：Su Yun bacillus：Bacillus amyloliquefaciens=2：1：1),
It is 4% commercially available bacillus pumilus by inoculum concentration：2% Su Yun bacillus：2% bacillus amyloliquefaciens H47 takes access to ferment respectively
Stoste, 120r/min oscillation and fermentation 6d, pH value 7.0,25 DEG C of temperature.
(5) coarse filtration is first carried out, sieves number is 20~32 mesh.Then thin filtering, sieves number is 80~100
Mesh, produces the insecticide that kuh-seng is fermentation raw material.
Embodiment 3
A kind of method that insecticide is prepared by fermenting raw materials of kuh-seng, comprises the following steps：
(1) the selection measured kuh-seng of fresh, matter is as fermentation primary raw material.
(2) pure water clear water is washed twice, is finally soaked, then cold sterilization drains, filled with 0.9% NaCl solution again
Enter in the cleaned container of pure water, probably account for the 40% of volume, add the house refuse that accounts for total liquid volume 15% and 1%
Starchy material is that fermenation raw liquid is standby.
(3) LB culture mediums are prepared, original is preserved into bacterial strain activation, the zymophyte that 120r/min shaken cultivations are obtained for about 48 hours
Liquid is standby；
(4) according to different inoculative proportion (commercially available bacillus pumilus：Su Yun bacillus：Bacillus amyloliquefaciens=1：2：
1), access is taken to send out respectively the commercially available bacillus pumilus of inoculum concentration 2%, 4% Su Yun bacillus and 2% bacillus amyloliquefaciens H47
Ferment stoste, 150r/min oscillation and fermentation 6d, pH value 7.0,25 DEG C of temperature.
(5) coarse filtration is first carried out, sieves number is 20~32 mesh.Then thin filtering, sieves number is 80~100
Mesh, produces the insecticide that kuh-seng is fermentation raw material.
Embodiment 4
A kind of method that insecticide is prepared by fermenting raw materials of kuh-seng, comprises the following steps：
(1) the selection measured kuh-seng of fresh, matter is as fermentation primary raw material.
(2) pure water clear water is washed twice, is finally soaked, then cold sterilization drains, filled with 0.9% NaCl solution again
Enter in the cleaned container of pure water, probably account for the 50% of volume, add the house refuse that accounts for total liquid volume 20% and 1%
Starchy material is that fermenation raw liquid is standby.
(3) LB culture mediums are prepared, original is preserved into bacterial strain activation, the zymophyte that 150r/min shaken cultivations are obtained for about 48 hours
Liquid is standby；
(4) according to different inoculative proportion (commercially available bacillus pumilus：Su Yun bacillus：Bacillus amyloliquefaciens=1：1：
2) it is, that 2% commercially available bacillus pumilus, 2% Su Yun bacillus and 4% bacillus amyloliquefaciens H47 take access respectively by inoculum concentration
Fermenation raw liquid, 200r/min oscillation and fermentation 6d, pH value 7.0,25 DEG C of temperature.
Coarse filtration is first carried out, sieves number is 20~32 mesh.Then thin filtering, sieves number is 80~100 mesh,
Produce the insecticide that kuh-seng is fermentation raw material.
Kuh-seng in the present invention contains abundant cometabolism material and kuh-seng ether extract and alcohol extract etc. to be had to plant
There are growth-promoting functions and drug effect is substantially, but be not limited only to this, the fresh weight or dry weight of kuh-seng according to actual conditions, can be determined
Addition or selection starchy material and domestic garbage mixing together, as long as insecticide reasonable mixture ratio, desinsection can be reached
The drug effect of agent can just play obvious effect.
The selection of other fermenation raw liquid and proportioning, connect bacterium amount and be also not necessarily limited to the combination that embodiment is lifted, can be according to actual feelings
Condition selects different combinations.
The preferred embodiments of the present invention are the foregoing is only, are not intended to limit the invention, all thinkings in the present invention
Within principle, any modification, equivalent substitution and improvements done etc. should be included in the scope of the protection.
Claims (9)
1. a kind of technology that insecticide is prepared by fermenting raw materials of kuh-seng, is characterized in using the measured kuh-seng of matter as main proferment
Material, adds starch and house refuse, using commercially available bacillus pumilus, Su Yun bacillus and bacillus amyloliquefaciens H47, production system
For insecticide, its technique is：
1) surface contaminants are cleaned with running water from the fresh kuh-seng without lesion, reuses Wuxi a century Shang Shui drinks Co., Ltd
The century-old still water clear water of offer is washed twice, is finally invaded and steeped 120 seconds with 0.9% NaCl solution again, drained, be put into tank, by body
Product is than being prince's-feather：House refuse：Starchy material:Pure water=4:1~2:0.01:4~5 are configured to fermenation raw liquid.
2) LB culture mediums are prepared, original is preserved into bacterial strain activation, the bacterium solution obtained for about 48 hours is cultivated standby；Again by commercially available short and small bud
Spore bacillus, Su Yun bacillus and bacillus amyloliquefaciens H47 access fermenation raw liquid according to a certain percentage
3) in fermentation system, the strain of system 4% is added, fermentation temperature is 25~35 DEG C, and rotating speed is 100~200rpm, is kept
PH=5~8,6~9d of fermentation time.
4) zymotic fluid after the completion of) fermenting is first through coarse filtration, and sieves number is 20~32 mesh；Thin filtering, sieves number again
For 80~100 mesh, you can prepare the insecticide using prince's-feather as raw material.
2. the method that a kind of kuh-seng that ferments according to claim 1 prepares insecticide, it is characterized in that：According to inoculative proportion
4%, commercially available bacillus pumilus, Su Yun bacillus and bacillus amyloliquefaciens H47 grade ratios are accessed into fermenation raw liquid, each bacteria concentration is
106CFU/ml, rotating speed is 100~200rpm, and ferment 3~9d, pH value 5.0~8.0,25 DEG C~35 DEG C of temperature.
3. the method that a kind of kuh-seng that ferments according to claim 1 prepares insecticide, it is characterized in that：The LB culture mediums of preparation
For：Tryptone 10g, yeast extract 5g, sodium chloride 10g, constant volume to 1L adjust pH=7.0.
4. the method that a kind of kuh-seng that ferments according to claim 1 prepares insecticide, it is characterized in that：Fermenation raw liquid volume ratio
For prince's-feather：House refuse：Starchy material:Pure water=4:1~2:0.01:4~5.
5. the method that a kind of kuh-seng that ferments according to claim 1 prepares insecticide, it is characterized in that：Fermentation strain is used
It is CCTCC M's 2013283 to be stored in the deposit number of China typical culture collection center on June 24th, 2013
Bacillus amyloliquefaciens H47 Bacillus amyloliquefaciens strain is fermentation strain.
6. the method that a kind of kuh-seng that ferments according to claim 1 prepares insecticide, it is characterized in that：The addition of strain is
The 4% of system.
7. the method that a kind of kuh-seng that ferments according to claim 1 prepares insecticide, it is characterized in that：The processing method of raw material
To clean surface contaminants from the fresh kuh-seng running water without lesion, bubble is invaded with 0.9% NaCl solution 120 seconds, drained standby
With.
8. the method that a kind of kuh-seng that ferments according to claim 1 prepares insecticide, it is characterized in that：Raw material and strain, life
Rubbish and starchy material living are well mixed, and fermentation temperature is 25~35 DEG C, 100~200rpm of rotating speed, keep pH=5~8, hair
6~9d of ferment time.
9. the method that a kind of kuh-seng that ferments according to claim 1 prepares insecticide, it is characterized in that：Hair after the completion of fermentation
Zymotic fluid is first through coarse filtration, and sieves number is 20~32 mesh；Thin filtering again, sieves number is 80~100 mesh.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109329323A (en) * | 2018-10-31 | 2019-02-15 | 百士泰（武汉）生物科技有限公司 | A kind of draft ferment insecticide and its preparation method and application |
| CN109430300A (en) * | 2018-11-21 | 2019-03-08 | 河南科技学院 | A kind of microbial fermentation pesticide and its preparation method and application |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1265834A (en) * | 1999-03-07 | 2000-09-13 | 宛吉斌 | Process for preparing natural plant biological biocide and biological fertilizer |
| CN101697740A (en) * | 2009-04-04 | 2010-04-28 | 冯子龙 | Novel traditional Chinese medicine pesticide and production technology thereof |
| CN103451128A (en) * | 2013-07-12 | 2013-12-18 | 江南大学 | Bacillus amyloliquefaciens and application thereof in preventing and controlling peach bleeding disease |
-
2017
- 2017-04-14 CN CN201710243507.3A patent/CN106967754A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1265834A (en) * | 1999-03-07 | 2000-09-13 | 宛吉斌 | Process for preparing natural plant biological biocide and biological fertilizer |
| CN101697740A (en) * | 2009-04-04 | 2010-04-28 | 冯子龙 | Novel traditional Chinese medicine pesticide and production technology thereof |
| CN103451128A (en) * | 2013-07-12 | 2013-12-18 | 江南大学 | Bacillus amyloliquefaciens and application thereof in preventing and controlling peach bleeding disease |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109329323A (en) * | 2018-10-31 | 2019-02-15 | 百士泰（武汉）生物科技有限公司 | A kind of draft ferment insecticide and its preparation method and application |
| CN109430300A (en) * | 2018-11-21 | 2019-03-08 | 河南科技学院 | A kind of microbial fermentation pesticide and its preparation method and application |
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