CN106867946A - A kind of method for preparing insecticide through microbial fermentation as raw material with Celastrus angulatus - Google Patents
A kind of method for preparing insecticide through microbial fermentation as raw material with Celastrus angulatus Download PDFInfo
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- CN106867946A CN106867946A CN201710243532.1A CN201710243532A CN106867946A CN 106867946 A CN106867946 A CN 106867946A CN 201710243532 A CN201710243532 A CN 201710243532A CN 106867946 A CN106867946 A CN 106867946A
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- celastrus angulatus
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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
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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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- Pest Control & Pesticides (AREA)
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Abstract
The invention provides a kind of technology for preparing biological source insecticide.Its process characteristic is that, with Celastrus angulatus as raw material, using being stored in China typical culture collection center, deposit number is：CCTCCNO：Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) H47 of M2013283 and commercially available bacillus pumilus and commercially available Dipel mixed fermentation.Raw material adds strain 100mL after suitably smashing, and add water 10L, 25 DEG C of -30 DEG C of 3 12d of fermentation, and finished product is obtained after filtering.Preparation method of the present invention and during non-pollution safety reliability, fermentation time cycle is short, process procedure is few, simple to operation, can reduce investment outlay and operating cost.Compared with traditional chemical medicament, its tool green safety, the advantage such as environment-friendly, and Promoting bacteria also has preferable disinsection prophylaxis and growth-promoting functions in itself, gathers around and has broad application prospects.
Description
Technical field
The present invention relates to solve one plant of bacillus amyloliquefacienses (Bacillusamyloliquefaciens) H47 and short and small bud
Spore bacillus and bacillus thuringiensis microbiological fermentation Celastrus angulatus prepare the production method of plant insecticide, belong to microorganism and fermentation
Technical field.
Background technology
Celastrus angulatus (Celastrus angulatus) is Celastraceae celastraceae plantses, and it is rattan shape shrub；Sprig often has
4-6 vertical ribs, the close life of hole skin, circle to ellipse, white, axillary bud oval shape, 2-4mm long.Ye great, nearly keratin, rectangular wealthy ellipse
Shape, wealthy avette, circle, 7-17cm long, 5-13cm wide, tip circle are wealthy, and center tool tip, lateral vein 5-7 pairs is substantially dashed forward on blade face
Rise, two sides is smooth or is leaner than on the master arteries and veins of blade back and has pubescence；Petiole 1.5-3cm long；Stipule is thread, caducous.Poly- umbrella circular cone
Inflorescence basidixed, bottom branch is longer than top branch, and slightly in tower taper, 10-20cm long, rachis and rachilla are smooth or by rust
Undercoat；Pedicel is shorter, and joint is at top；Calyx valvate is arranged, and triangle is about 1.2mm, nearly full edge to avette；Petal
Rectangle, is about 2mm, and about 1.2mm wide, edge is irregular；Floral disc meat, shallow discoid or plate-like, 5 shallow split；Stamen raw floral disc
Under, 3mm is about, staminodium is about 1mm in female flower；Gynoecium 3-4mm long, ovary is spherical, and column cap contrary flexure is moved back in male flower
Change gynoecium and be about 1.2mm.Capsule is closely spherical, diameter 8-10mm；Seed ellipticity, 3.5-5.5mm long, diameter 1.5-3mm.Florescence
The 5-5 months.It is distributed widely in the hills and mountain area of China the Yellow River, the Yangtze river basin.
The root skin and stem skin of Celastrus angulatus contain various strength insecticidal constituents, separate identification from root skin or seed at present
Go out dozens of noval chemical compound, particularly obtain 4 crystallizations from kind of oil, be i.e. Celastrus angulatus ester I- IV, 5 pure days obtained from root skin
Right product, i.e. Celastrus angulatus I-V.Active insecticidal components in these Celastrus angulatus are referred to as Celastrus angulatus；Wherein Celastrus angulatus I
There are Antifeedant Effects to insect, Celastrus angulatus II, III have stomach toxicity toxic action, bitter skin to insects such as black cutworm, lopper worms
Rattan toxin IV there is Selective anesthesia to act on insect, and Celangulin V acts on the digestive system of insect, and intestines intestinal wall is thin in destruction
The plasma membrane and inner membrance body fluid of born of the same parents enter alimentary canal through intestinal wall cell, cause insect to be suffered from vomiting and diarrhoea, dehydration and it is dead, thus be used as
Prepare botanical pesticide.
Plant produces in evolution has the secondary metabolites of protective effect, and these materials can often resist other lifes
The infringement of thing.Typically easily degraded with plant time plant insecticide of the metabolin as main active, will not to soil with
Water source forms persistence influence；Plant insecticide is general relatively low to fish, bird, mammal and people's toxicity, uses safety：Plant
Insecticide has certain selection index system to natural enemy, it is possible to control the rampant again of insect pest.Above-mentioned advantage causes that plant source is killed
Worm agent turns into replacement chemical pesticide control insect and selects well.Using suitable plant growth-promoting bacteria fermentation Celastrus angulatus, do not only have
Beneficial to the extraction of physiological activator, it is also less prone to cause the inactivation of active material, Promoting bacteria also has preferable disinsection prophylaxis in itself
And growth-promoting functions.Be conducive to improving plant growth and enhancing insecticidal effect, be conducive to environmental protection and Celastrus angulatus plant in itself
Effectively utilize.
The content of the invention
In view of the foregoing defects the prior art has, the present invention was raw material using Celastrus angulatus plant, with June 24th, 2013
China typical culture collection center is stored in, deposit number is：CCTCCNO：The bacillus amyloliquefaciens of M2013283
(bacterial strain is hereinafter referred to as (Bacillus amyloliquefaciens) H47：Bacillus amyloliquefaciens
H47CCTCCM2013283) and commercially available bacillus pumilus and commercially available Dipel fermentation prepare plant insecticide, favorably
In plant growth and enhancing insecticidal effect is improved, be conducive to environmental protection.
Fermentation technique scheme of the invention：
1) strain prepares：By bacillus amyloliquefaciens H47CCTCCM2013283 and commercially available bacillus pumilus, commercially available Soviet Union
Cloud gold bacillus is inoculated in seed culture medium respectively, in 30 DEG C, 250r/min concussion and cultivates 12h；The seed culture based component is pressed
G/L is calculated as：Peptone 10, yeast extract 5, NaCl 10, remaining composition is water, and 121 DEG C of autoclaving 15min adjust pH to 7.0.
Seed is done in mixing by a certain percentage.
2) zymotic fluid prepares：From superior in quality Celastrus angulatus, rinsed with water, 70% -75% alcohol-pickled 3min, used
Water is cleaned, stand-by after being blended through pulper.
3) fermentation ullage disappears and air duct sterilization：Fermentation ullage is carried out by instructions book to disappear.
4) zymotic fluid disappears in fact：Step 2 gained Celastrus angulatus is put into fermentation tank, supplement running water to final volume is 10L, is pressed
Instructions book carries out real elimination bacterium.
5) inoculated and cultured：When fermentation jar temperature is down to 37 DEG C, by step 1 gained seed by volume 1% inoculum concentration,
It is seeded in the fermentation tank equipped with Celastrus angulatus, is cultivated 3-12 days under the conditions of 25-30 DEG C, the throughput of technological requirement is set and is protected
Tank pressure is held, each other culture parameters cultures are set.
6) discharge：Culture is finished, and releases zymotic fluid, and filtering, zymotic fluid obtained by after filtering is insecticide finished product.
Beneficial effects of the present invention：
A kind of method of biological source insecticide is obtained the invention provides fermentation Celastrus angulatus.Preparation method of the present invention and process
Middle non-pollution safety reliability, fermentation time cycle is short, process procedure is few, simple to operation, can reduce investment outlay and operating cost.With
Traditional chemical medicament is compared, its tool green safety, the advantage such as environment-friendly, and Promoting bacteria also have in itself preferable disinsection prophylaxis and
Growth-promoting functions, gather around and have broad application prospects.
Specific implementation method：
With reference to embodiment, the present invention is described further.Following examples are only several specific realities of the invention
Example is applied, but design concept of the invention is not limited thereto, all changes for carrying out unsubstantiality to the present invention using this design,
The behavior for invading the scope of the present invention should be belonged to.
Embodiment 1
1) strain prepares：By bacillus amyloliquefaciens H47CCTCCM2013283 and commercially available bacillus pumilus, commercially available Soviet Union
Cloud gold bacillus is inoculated in seed culture medium respectively, in 30 DEG C, 250r/min concussion and cultivates 12h；The seed culture based component is pressed
G/L is calculated as：Peptone 10, yeast extract 5, NaCl 10, remaining composition is water, and 121 DEG C of autoclaving 15min adjust pH to 7.0.
By 1:1:Seed is done in 1 mixing.
2) zymotic fluid prepares：From superior in quality Celastrus angulatus, rinsed with water, 70% -75% alcohol-pickled 3min, used
Water is cleaned, stand-by after being blended through pulper.
3) fermentation ullage disappears and air duct sterilization：Fermentation ullage is carried out by instructions book to disappear.
4) zymotic fluid disappears in fact：Step 2 gained Celastrus angulatus is put into fermentation tank, supplement running water to final volume is 10L, is pressed
Instructions book carries out real elimination bacterium.
5) inoculated and cultured：When fermentation jar temperature is down to 37 DEG C, by step 1 gained seed by volume 1% inoculum concentration,
It is seeded in the fermentation tank equipped with Celastrus angulatus, is cultivated 3 days under the conditions of 25 DEG C, the throughput of technological requirement is set and tank pressure is kept,
Each other culture parameters cultures are set.
6) discharge：Culture is finished, and releases zymotic fluid, and filtering, zymotic fluid obtained by after filtering is insecticide finished product.
Embodiment 2
1) strain prepares：By bacillus amyloliquefaciens H47CCTCCM2013283 and commercially available bacillus pumilus, commercially available Soviet Union
Cloud gold bacillus is inoculated in seed culture medium respectively, in 30 DEG C, 250r/min concussion and cultivates 12h；The seed culture based component is pressed
G/L is calculated as：Peptone 10, yeast extract 5, NaCl 10, remaining composition is water, and 121 DEG C of autoclaving 15min adjust pH to 7.0.
By 1:1:Seed is done in 1 mixing.
2) zymotic fluid prepares：From superior in quality Celastrus angulatus, rinsed with water, 70% -75% alcohol-pickled 3min, used
Water is cleaned, stand-by after being blended through pulper.
3) fermentation ullage disappears and air duct sterilization：Fermentation ullage is carried out by instructions book to disappear.
4) zymotic fluid disappears in fact：Step 2 gained Celastrus angulatus is put into fermentation tank, supplement running water to final volume is 10L, is pressed
Instructions book carries out real elimination bacterium.
5) inoculated and cultured：When fermentation jar temperature is down to 37 DEG C, by step 1 gained seed by volume 1% inoculum concentration,
It is seeded in the fermentation tank equipped with Celastrus angulatus, is cultivated 6 days under the conditions of 27 DEG C, the throughput of technological requirement is set and tank pressure is kept,
Each other culture parameters cultures are set.
6) discharge：Culture is finished, and releases zymotic fluid, and filtering, zymotic fluid obtained by after filtering is insecticide finished product.
Embodiment 3
1) strain prepares：By bacillus amyloliquefaciens H47CCTCCM2013283 and commercially available bacillus pumilus, commercially available Soviet Union
Cloud gold bacillus is inoculated in seed culture medium respectively, in 30 DEG C, 250r/min concussion and cultivates 12h；The seed culture based component is pressed
G/L is calculated as：Peptone 10, yeast extract 5, NaCl 10, remaining composition is water, and 121 DEG C of autoclaving 15min adjust pH to 7.0.
By 1:1:Seed is done in 1 mixing.
2) zymotic fluid prepares：From superior in quality Celastrus angulatus, rinsed with water, 70% -75% alcohol-pickled 3min, used
Water is cleaned, stand-by after being blended through pulper.
3) fermentation ullage disappears and air duct sterilization：Fermentation ullage is carried out by instructions book to disappear.
4) zymotic fluid disappears in fact：Step 2 gained Celastrus angulatus is put into fermentation tank, supplement running water to final volume is 10L, is pressed
Instructions book carries out real elimination bacterium.
5) inoculated and cultured：When fermentation jar temperature is down to 37 DEG C, by step 1 gained seed by volume 1% inoculum concentration,
It is seeded in the fermentation tank equipped with Celastrus angulatus, is cultivated 9 days under the conditions of 30 DEG C, the throughput of technological requirement is set and tank pressure is kept,
Each other culture parameters cultures are set.
6) discharge：Culture is finished, and releases zymotic fluid, and filtering, zymotic fluid obtained by after filtering is insecticide finished product.
Claims (7)
1. a kind of method that Celastrus angulatus that ferments prepares biological source insecticide.It is characterized in that with Celastrus angulatus as raw material, addition strain hair
Ferment prepares insecticide, and the method is comprised the following steps：
1) strain prepares：By bacillus amyloliquefaciens H47CCTCCM2013283 and commercially available bacillus pumilus, commercially available Su Yun gold
Bacillus is inoculated in seed culture medium respectively, in 30 DEG C, 250r/min concussion and cultivates 12h；The seed culture based component is based on g/L
For：Peptone 10, yeast extract 5, NaCl 10, remaining composition is water, and 121 DEG C of autoclaving 15min adjust pH to 7.0.By one
Seed is done in certainty ratio mixing.
2) zymotic fluid prepares：From superior in quality Celastrus angulatus, rinsed with water, 70% -75% alcohol-pickled 3min is washed with water
Only, it is stand-by after being blended through pulper.
3) fermentation ullage disappears and air duct sterilization：Fermentation ullage is carried out by instructions book to disappear.
4) zymotic fluid disappears in fact：Step 2 gained Celastrus angulatus is put into fermentation tank, supplement running water to final volume is 10L, by correlation
Specification carries out real elimination bacterium.
5) inoculated and cultured：When fermentation jar temperature is down to 37 DEG C, by step 1 gained seed by volume 1% inoculum concentration, inoculation
Into the fermentation tank equipped with Celastrus angulatus, cultivated 3-12 days under the conditions of 25-30 DEG C, the throughput of technological requirement is set and tank is kept
Pressure, sets each other culture parameters cultures.
6) discharge：Culture is finished, and releases zymotic fluid, and filtering, zymotic fluid obtained by after filtering is insecticide finished product.
2. the method that fermentation Celastrus angulatus according to claim 1 prepares biological source insecticide, it is characterized in that：The strain list
As fermentation seed after solely seed culture medium culture 12h mixes by a certain percentage.
3. the method that fermentation Celastrus angulatus according to claim 1 prepares biological source insecticide, it is characterized in that：Strain (Xie Dian
Afnyloliquefaciens H47CCTCCM2013283 and commercially available bacillus pumilus, commercially available Dipel) use seed culture based formulas
It is (g/L)：Peptone 10, yeast extract 5, NaCl 10, remaining composition is water, and 121 DEG C of autoclaving 15min adjust pH to 7.0.
4. the method that fermentation Celastrus angulatus according to claim 1 prepares biological source insecticide, it is characterized in that：Celastrus angulatus treatment
Method is：From superior in quality Celastrus angulatus, rinsed with water, 70% -75% alcohol-pickled 3min is washed, through pulper
Blend rear stand-by.
5. the method that fermentation Celastrus angulatus according to claim 1 prepares biological source insecticide, it is characterized in that：The addition of strain
Measure is the 1% of fermentation system.
6. the method that fermentation Celastrus angulatus according to claim 1 prepares biological source insecticide, it is characterized in that：Raw material and strain
It is well mixed, 25-30 DEG C of fermentation temperature, fermentation time 3-12d.
7. the method that fermentation Celastrus angulatus according to claim 1 prepares biological source insecticide, it is characterized in that：Fermentation time week
Phase is short, and process procedure is few, and the insecticide of preferable disinsection prophylaxis and growth-promoting functions can be obtained after fermentation.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108464317A (en) * | 2018-03-15 | 2018-08-31 | 郝之奎 | A kind of method that peppermint prepares mosquito-repellent with lemongrass fermentation |
| CN108513993A (en) * | 2018-03-15 | 2018-09-11 | 郝之奎 | A method of preparing insecticide by fermenting raw materials of the root of Beijing euphorbia |
| CN110200028A (en) * | 2019-06-05 | 2019-09-06 | 广西天浩农业发展有限公司 | A kind of cane planting insecticide and preparation method thereof |
| CN112514917A (en) * | 2020-12-03 | 2021-03-19 | 黑龙江沪生农业发展有限公司 | Composite microbial pesticide and preparation method thereof |
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| CN1452869A (en) * | 2002-04-26 | 2003-11-05 | 郭立卿 | Prepn of wide-spectrum biopesticide of Chinese herbal medicine |
| CN101124916A (en) * | 2007-08-06 | 2008-02-20 | 大连奥德植保药业有限公司 | Biological pesticide mixed with plant insecticide sterilization component and amino-oligosaccharin and preparation method thereof |
| CN101697740A (en) * | 2009-04-04 | 2010-04-28 | 冯子龙 | Novel traditional Chinese medicine pesticide and production technology thereof |
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2017
- 2017-04-14 CN CN201710243532.1A patent/CN106867946A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1452869A (en) * | 2002-04-26 | 2003-11-05 | 郭立卿 | Prepn of wide-spectrum biopesticide of Chinese herbal medicine |
| CN101124916A (en) * | 2007-08-06 | 2008-02-20 | 大连奥德植保药业有限公司 | Biological pesticide mixed with plant insecticide sterilization component and amino-oligosaccharin and preparation method thereof |
| CN101697740A (en) * | 2009-04-04 | 2010-04-28 | 冯子龙 | Novel traditional Chinese medicine pesticide and production technology thereof |
Non-Patent Citations (1)
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| 李衍方 等: "杀虫植物苦皮藤的研究新进展", 《农药》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108464317A (en) * | 2018-03-15 | 2018-08-31 | 郝之奎 | A kind of method that peppermint prepares mosquito-repellent with lemongrass fermentation |
| CN108513993A (en) * | 2018-03-15 | 2018-09-11 | 郝之奎 | A method of preparing insecticide by fermenting raw materials of the root of Beijing euphorbia |
| CN110200028A (en) * | 2019-06-05 | 2019-09-06 | 广西天浩农业发展有限公司 | A kind of cane planting insecticide and preparation method thereof |
| CN112514917A (en) * | 2020-12-03 | 2021-03-19 | 黑龙江沪生农业发展有限公司 | Composite microbial pesticide and preparation method thereof |
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