CN105638651B - A kind of preparation method of atrophy bacillus wettable powder - Google Patents
A kind of preparation method of atrophy bacillus wettable powder Download PDFInfo
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- CN105638651B CN105638651B CN201510951083.7A CN201510951083A CN105638651B CN 105638651 B CN105638651 B CN 105638651B CN 201510951083 A CN201510951083 A CN 201510951083A CN 105638651 B CN105638651 B CN 105638651B
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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
- 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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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05C—NITROGENOUS FERTILISERS
- C05C11/00—Other nitrogenous fertilisers
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
Abstract
The present invention relates to biological pesticide technical field, specifically disclosed is a kind of wettable powder of atrophy bacillus containing gemma germinant and preparation method thereof, and the raw material of the wettable powder is made up of following composition by weight：40 70 parts of atrophy fermentation of bacillus liquid；Filler：20 30 parts of kaolin；Wetting agent：14 parts of neopelex；Dispersant：425 0.2 1 parts of Morwet D；Gemma germinant：0.1 1 parts of fructose, 0.1 1 parts of threonine, 2,6 0.1 0.5 parts of pyridinedicarboxylic acid calcium；Uv-protector：12 parts of trehalose, 0.2 1 parts of carboxymethyl cellulose；Drying protectant：15 parts of sucrose, 0.2 1 parts of sodium glutamate.Gemma speed of germinating is fast after atrophy bacillus wettable powder prepared by the present invention applies crop field, and germination rate is high, can significantly improve its prevention effect, can reduce gemma dosage, reduces production cost.
Description
Technical field
The present invention relates to microbial pesticide technical field, and in particular to a kind of atrophy gemma containing atrophy gemma germinant
Bacillus wettable powder and preparation method thereof.
Background technology
Atrophy bacillus (Bacillus atrophaeus, non-pathogenic bacteria, a kind of mutation of bacillus subtilis,
Solvable black colonies can be formed on culture medium containing carbohydrate, it is mainly isolated from soil.In agricultural, work
The fields such as industry, scientific research, medical treatment and health are all widely used.Atrophy bacillus can produce catalase, do not produce and connect
Catalase, VP reacting positives, hydrogen sulfide, indoles and dihydroxyacetone are not produced, be able to can be hydrolyzed by nitrate reduction into nitrite
Starch and casein.Atrophy bacillus optimum growth temperature scope：28~30℃；Maximum growth temperature range：50~55℃；It is minimum
Growth temperature range：5~10℃.Atrophy bacillus can be 4-9 in PH and be grown in 7%NaCl.Atrophy bacillus energy
Utilize the sour not aerogenesis of L-arabinose, D-Fructose, D-Glucose, mannitol, salicin, sucrose, trehalose and xylose production, it is impossible to
Utilize melibiose and lactose fermentation.Luo Huadong et al. is filtered out to the strong pathogenic bacterium biocontrol bacterial strain atrophy of colorado potato bug tool
Bacillus CPB072, CPB108, can degrade chitin, protein, can utilize sucrose, mannose ferment, it is impossible to carry out D- Portugals
Grape sugar fermentation, nitrate reduction can be sold hydrochlorate by CPB072 into Asia, but CPB108 can not be by nitrate reduction into nitrite.
1872, in the first division bacteria system established by Germany scientist Cohn, built according to ne ar feature
Bacillus (BaciUus) is stood.Bacillus is a kind of Gram-positive that can produce spherical or oval gemma
Bacterium, to the bad factor resistance such as arid, ultraviolet, low temperature, high temperature, distribution is wide, be present in water, soil, air, plant and
In animal intestinal tract.Equally, atrophy bacillus is distributed more widely in the environment.Han Song et al. is isolated in small bell tattooing portion organ
One plant of endogenetic bacteria HS-4, the bacterium are ultimately determined to atrophy bacillus.Xue Peng volume is from the Qinghai-Tibet of more than 4000 meters of mean sea level
Atrophy bacillus is separated in the soil sample of plateau.Fall et al. has found atrophy bacillus in root system of plant be present.
Biological control (biological control) is to utilize the correlation between biological species, and it is another to reach suppression
Or the method for another kind of harmful organism.Microorganism Biocontrol Effect, it can be held as a kind of important method of biological control, and one kind
The means of continuous control disease, receive much concern.Recently, people isolate atrophy bacillus inside soil and crops, gradually
Use it for controlling disease.
Wuzhong is red to being identified with 2 plants of atrophy bud pole bacteriums of salt tolerant growth-promoting and prophylaxis effect, and studies itself and its
Its 3 plants of bacterial strain mixed culture is acted on observing the preventive effect of mix bacterium agent.Apply rainbow and one plant is isolated and purified out from soil with Huang
The atrophy bacillus of bent clouds antagonistic activity, suppress aspergillus flavus growth result using the fermented supernatant fluid observation of the bacterial strain.
Shi Lei etc. is screened from a large amount of biocontrol microorganisms by flat board transparent circle method to be obtained one plant and has the thin of chitinase activity
Bacteria strain CAB-1, the bacterial strain show the various plants such as botrytis cinerea disease fungus stronger antagonistic activity.Pass through physiology
Biochemical, 16S rDNA and gyrB gene sequencings, atrophy bacillus (Bacillus is accredited as by bacterial strain CAB-1
atrophaeus).1 plant of the separation screening atrophy bacillus to aspergillus flavus with antagonism from soil such as Liu Ding
(BaciUm atrophaeus, further further investigation.From nutrient competition, directly parasitism, induction host resistance and antibiosis
Many aspects inquire into the action pathway that this plant of Antagonistic Fungi suppresses aspergillus flavus, and host resistance research is acted on and induced by nutrient competition
It was found that the cometabolism that may contain one or more of atrophy bacillus in atrophy bacillus nutrient solution sterile supernatant produces
Thing, there is antagonism aspergillus flavus.
Bacillus as biological prevention and control agent based on wettable powder, be although easy to store and transport by this formulation at present,
But many restrictive factors in production application be present.As administration wettable powder in major part using gemma as weigh product in
The unitary basis of active ingredient, if gemma can not be sprouted in time, application effect is then bad, as Wang Qian utilizes bacillus subtilis
When TR21 prevents and treats Brazilian any of several broadleaf plants droop, pre-stage test can be prevented and treated effectively using TR21 bacterial strains NB fermentation liquid irrigating roots and axil inoculation
Droop, but the pouring root that wettable powder is prepared using gemma is undesirable with axil effect, demonstrates after removing zymotic fluid
Pure gemma may be because that sprouting is difficult and declines preventive effect.
Bacillus is widely distributed in nature, and when external condition is adapted to growth, brood cell can be passed through by resting state
A series of complex biochemical reactions grow into trophosome and recover metabolism, and this process is referred to as sprouting
(germination).Gemma is sprouted and influenceed by exogenous nutrition material in nature, and this kind of nutriment is referred to as " sprouting
Agent ", usual nutrition induction gemma sprout the single effect for including amino acid, carbohydrate and fast cry of certain animals nucleosides, also include their collaboration
Effect (such as：AGFK；A：The asparagus fern phthalein amine of L mono-；G：Glucose；F：Fructose；K：Potassium ion).When nutriment touches gemma
When, a series of changes can occur for gemma inside,（1）, H inside gemma coreTenAnd Zn2+Discharge, cause gemma core internal pH by
Faintly acid rises to alkalescent；(2), Ca inside gemma core2+-- DPA a large amount of releases；(3), gemma core is hydrated, internal Ca2+--
DPA is substituted by water, and the heat resistance of gemma declines；(4), the gemma skin glycan layer hydrolysis of cortex, gemma swelling of nucleus；(5)、
Gemma core is further hydrated, and gemma core internal protein mobility enhancing, enzymatic activity is strengthened, and synthesizes ATP；(6), degraded SASP eggs
In vain, to discharge gemma DNA, and then the synthesis of RNA, protein, DNA in gemma growth course are guided.
After gemma and the gemma germinant for promoting to sprout mix within the several seconds, germinant is quickly across gemma clothing and cortex
The surface of intine is reached, is interacted with the receptor protein on intine, is excited gemma to enter germinating, connect
And start a succession of sprouting event, germination process afterwards will not stop because removing germinant.
At this stage, there is not also the technical scheme in atrophy bacillus wettable powder to disclose, do not have in atrophy gemma yet
The report of the research of gemma germinant is added in bacillus wettable powder.
Further, since the compatibility of the auxiliary agent such as different bacillus and wetting agent, dispersant, carrier is to all kinds of gemma bars
Bacterium adsorptivity is different, and the formula of existing bacillus wettable powder or preparation technology are not used to prepare addition bud
The atrophy bacillus wettable powder of spore germinant, the effective adsorbance of bacterium in wettable powder, survival rate, to harmful bacteria
The stability difference of prevention effect and wettable powder dosage is larger, and the preparation method of its whole wettable powder must give overall consideration to
Each carrier adsorption ability, to protective effect in the drying process of atrophy bacillus, wetting agent, dispersant is to wettable powder
Suspensibility, the influence of MEBO ribbon gauze, and comprehensive screening is carried out to gemma germinant just to access MEBO ribbon gauze short, suspend
Rate is high, and the gemma holding time is long, using rear gemma energy fast-germination, the good atrophy bacillus wettable powder of preventive effect.
The content of the invention
The technical problems to be solved by the invention are：For above-mentioned the deficiencies in the prior art, there is provided one kind is sprouted containing gemma
Atrophy bacillus wettable powder of agent and preparation method thereof is sent out, gemma germinant can make atrophy bacillus wettable powder
After incorporating water, atrophy gemma energy fast-germination, the germination rate of atrophy gemma is greatly improved, the atrophy bacillus that it is prepared is wettable
Property pulvis spore content be not less than 10,000,000,000 CFU/ grams, suspensibility is not less than 82%, and wetting time is not higher than 75S, finess disintegrating
400 mesh sieves are crossed, moisture is not higher than 5%, and preparation method is stable.
In order to achieve the above object, a kind of atrophy bacillus wettable powder is made up of following parts by weight：Atrophy gemma
Bacillus fermentation liquid 40-70 parts；Filler：Kaolin 20-30 parts；Wetting agent：Neopelex 1-4 parts；Dispersant：
Morwet D-425 0.2-1 parts；Gemma germinant：Fructose 0.1-1 parts, threonine 0.1-1 parts, 2, dipicolimic acid 2 calcium
0.1-0.5 parts；Uv-protector：Trehalose 1-2 parts, carboxymethyl cellulose 0.2-1 parts；Drying protectant：Sucrose 1-5 parts, paddy
Propylhomoserin sodium 0.2-1 parts.
A kind of preparation method of atrophy bacillus wettable powder, is made up of suddenly the following steps：
First, the preparation of atrophy fermentation of bacillus liquid
Culture presevation pipe is taken out, flat board is drawn with nutrient broth solid medium and is recovered, 30 DEG C are cultivated 48 hours.Flat
Picking single bacterium colony is seeded in 50 milliliters of nutrient broth mediums under plate, 30 DEG C of concussion and cultivates 24 hours in incubator.Kind
Son uses 5% inoculum concentration, the atrophy bacillus liquid culture medium being seeded in the big triangular flasks of 5L, liquid amount 2L, 30 DEG C of shakes
Culture 16-28 hours are swung, detect its bacterial concentration, bacterium solution content is more than 3,000,000,000/milliliter, you can as atrophy bacillus seed
Liquid；Atrophy bacillus seed liquor is seeded in 1000L fermentation tank, liquid amount be 600L fermentation medium, throughput
It was that liquid-gas ratio per minute is 1 for first 16 hours：0.8, throughput is 1 thereafter：1；24 hours are 30 DEG C before cultivation temperature, open and stir
Mix, rotating speed 160r/min, thereafter, temperature was improved 1 DEG C in every 2 hours, cultivate 26-32 hours, detect its spore content and exceed
10000000000 CFU/ml, you can terminate fermentation, as atrophy fermentation of bacillus liquid；
Wherein, the nutrient broth medium：Peptone 10g, beef extract 3g, sodium chloride 5g, water 1000mL, pH7.2, Gu
Agar 2% is added in body culture medium；Wherein, the atrophy bacillus liquid culture medium：Glucose 15g/L, the g/ of beef extract 10
L, the g/L of peptone 5, the g/L of fish meal 5, the g/L of magnesium sulfate 0.5, the g/L of manganese sulfate 0.5, the g/L of potassium dihydrogen phosphate 0.5, phosphoric acid hydrogen
The g/L of dipotassium 0.5, calcium chloride 0.5 g/L, pH7.2；
Wherein, described fermentation medium：Glucose 15g/L corn flour 10 g/L, the g/L of corn steep liquor 30, peptone 10
G/L, the g/L of magnesium sulfate 0.5, the g/L of manganese sulfate 0.5, the g/L of potassium dihydrogen phosphate 0.5, the g/L of dipotassium hydrogen phosphate 0.5, calcium chloride 0.5
G/L, calcium carbonate 5g/L, pH7.2；The condition of each medium sterilization is：0.10-0.15MPa, 121 DEG C sterilize 30 minutes；
2nd, by atrophy fermentation of bacillus liquid 40-70 parts；Filler：Kaolin 20-30 parts；Uv-protector：Trehalose 1-
2 parts, carboxymethyl cellulose 0.2-1 parts；Drying protectant：Sucrose 1-5 parts, sodium glutamate 0.2-1 parts；It is spray-dried after mixing,
Obtain atrophy bacillus pulvis；
3rd, by wetting agent：Neopelex 1-4 parts；Dispersant：Morwet D-425 0.2-1 parts；Gemma is sprouted
Send out agent：Fructose 0.1-1 parts, threonine 0.1-1 parts, 2, dipicolimic acid 2 calcium 0.1-0.5 parts；It is uniformly mixed so as to obtain atrophy bacillus
Auxiliary agent；
4th, the atrophy bacillus pulvis obtained by step 2 and step 3 and auxiliary agent are mixed and crossed and pass through air-flow crushing
Machine crushed 400 mesh sieves, detects its spore content and is not less than 10,000,000,000 cfu/g, suspensibility is not less than 82%, and wetting time is not higher than
75s, moisture are less than 5%, vacuum packaging, as atrophy bacillus wettable powder.
Wherein, atrophy bacillus wettable powder optimum weight part composition is：Atrophy fermentation of bacillus liquid 60
Part；Filler：28 parts of kaolin；Wetting agent：3 parts of neopelex；Dispersant：1 part of Morwet D-425；Gemma is sprouted
Send out agent：0.8 part of fructose, 0.4 part of threonine, 2,0.5 part of dipicolimic acid 2 calcium；Uv-protector：1.8 parts of trehalose, carboxylic first
0.8 part of base cellulose；Drying protectant：4 parts of sucrose, 0.8 part of sodium glutamate.
Gemma is sprouted and influenceed by exogenous nutrition material in nature, and this kind of nutriment is referred to as " germinant ", leads to
In the case of often, after the gemma germinant mixing of gemma and promotion sprouting within the several seconds, germinant is quickly across gemma clothing and skin
Layer reaches the surface of intine, is interacted with the receptor protein on intine, excites gemma to enter germinating,
Then a succession of sprouting event is started, germination process afterwards will not stop because removing germinant.Wettable powder of the present invention
In contain gemma germinant：Fructose, threonine, 2, dipicolimic acid 2 calcium；It is from glycosides propylhomoserin, alanine, valine, bright ammonia
Acid, isoleucine, serine, threonine, methionine, cystine, asparagine, glutamine, aspartic acid, glutamic acid, rely
Propylhomoserin, arginine, phenylalanine, tyrosine, tryptophan, histidine, proline, inosine, Dan Yin is carried out in 2,6 pyridine dicarboxyl calcium
Element experiment, and the germinant that there is obvious induced atrophy gemma to sprout that associativity is tested to obtain.Through overtesting, of the invention withers
Bacillus wettable powder contract with pressing 1：1000 ratio carries out wetting and is well mixed, and gemma germinant now can be fully
After being contacted with gemma, so as to promote it to sprout soon, through overtesting, after 30 minutes, gemma germination rate has reached more than 80%, and
Not plus gemma germinant atrophy gemma after 4 hours, its gemma germination rate has only reached 45% or so.
The present inventor passes through to carrier（Such as bentonite, talcum powder, kaolin, diatomite, white carbon, ground phosphate rock, lightweight carbon
Sour calcium）, auxiliary agent（Wetting agent and dispersant）（Polyvinyl alcohol, polyethylene glycol, CMC-Na, Tween-60, neopelex,
T-shaped lignin, lauryl sodium sulfate, Tea Saponin, Morwet EFW, Morwet D425, My100, D110, Arab's power glue,
2- naphthalene sulfonic acid formaldehyde polymer sodium salts, OPEO, diisooctyl Disodium sulfosuccinate, methylcellulose, wood
Quality sodium sulfonate）Filler（Diatomite, kaolin, bentonite, white carbon, precipitated calcium carbonate, talcum powder）, ultraviolet protective agent
（Ascorbic acid, dextrin, neopelex is Congo red, carboxymethyl cellulose, lecithin）, drying protectant（Defatted milk
Powder, sucrose, sodium glutamate, maltose, mannose, methylcellulose etc.）Screened, its every kind of auxiliary agent is to atrophy bacillus
Gemma survival, suspension, wettability tested, and atrophy bacillus is determined with orthogonal test finally by more wheel single factor tests
The formula of wettable powder.By checking, atrophy bacillus wettable powder of the invention preserves 1 year at 20-25 DEG C,
Its gemma survival rate also can reach 9,200,000,000 CFU/ grams.
The wettable powder of the present invention has adopted vacuum packaging, reduces contact of the gemma with air, reduces the machine of its sprouting
Rate.
The moisture of the wettable powder of the present invention is less than 5%, even if containing bud in the wettable powder of low water content
Spore germinant（Fructose, threonine, 2, dipicolimic acid 2 calcium）, to the gemma activity in its storage process also without obvious shadow
Ring, do not reduce its gemma in storage process.
Containing unimolecule amino acid such as threonines in wettable powder in the present invention, initial stage not only is being used as sprouting
Agent, and then can play a part of fertilizer efficiency as amino acid fertilizer after applying crop field.
The present invention compared with prior art, has advantages below and effect：
This is domestic first by the gemma germinant addition atrophy bacillus wettable agent progress industry of atrophy bacillus
Metaplasia is produced, and is greatly improved gemma and is sprouted speed and germination rate, is solved because gemma is sprouted slowly, is prevented caused by germination rate is low
Control the problem of effect is poor, and effect is unstable；
Directly prepared first using atrophy fermentation of bacillus liquid with the related wettable powder raw material such as carrier, filler
Wettable powder, reduce technological process, reduce the loss of atrophy bacillus gemma during wettable powder is prepared
Rate；
Providing a kind of spore content and be not less than 10,000,000,000 cfu/g, suspensibility is not less than 82%, and wetting time is not higher than 75s,
Moisture is less than 5%, the preparation method of vacuum-packed atrophy bacillus wettable powder.
Embodiment
Substantive distinguishing features of the present invention can be embodied from the following examples, but these embodiments are only used as explanation, without
It is to limit the invention.
Embodiment 1
A kind of atrophy bacillus wettable powder is made up of following parts by weight：60 parts of atrophy fermentation of bacillus liquid；Fill out
Material：28 parts of kaolin；Wetting agent：3 parts of neopelex；Dispersant：1 part of Morwet D-425；Gemma germinant：
0.8 part of fructose, 0.4 part of threonine, 2,0.5 part of dipicolimic acid 2 calcium；Uv-protector：1.8 parts of trehalose, carboxymethyl cellulose
0.8 part of element；Drying protectant：4 parts of sucrose, 0.8 part of sodium glutamate.
A kind of preparation method of atrophy bacillus wettable powder, is made up of suddenly the following steps：
First, the preparation of atrophy fermentation of bacillus liquid
Taking out culture presevation pipe, (atrophy bacillus Bacillus atrophaeus CICC 23590, are purchased in China
Research for Industrial Microbial Germ preservation administrative center, droop can be prevented and treated), draw flat board with nutrient broth solid medium and recovered,
30 DEG C are cultivated 48 hours.Picking single bacterium colony is seeded in 50 milliliters of nutrient broth mediums under flat board, 30 in incubator
DEG C concussion and cultivate 24 hours.Seed uses 5% inoculum concentration, the atrophy bacillus liquid culture being seeded in the big triangular flasks of 5L
Base, liquid amount 2L, 30 DEG C of concussion and cultivate 16-28 hours, detects its bacterial concentration, bacterium solution content is more than 3,000,000,000/milliliter, you can
As atrophy bacillus seed liquor；Atrophy bacillus seed liquor is seeded in 1000L fermentation tank, liquid amount 600L
Fermentation medium, throughput be first 16 hours be that liquid-gas ratio per minute is 1：0.8, throughput is 1 thereafter：1；Before cultivation temperature
24 hours are 30 DEG C, open stirring, rotating speed 160r/min, thereafter, temperature are improved into 1 DEG C in every 2 hours, cultivated 30 hours, detection
Its spore content is 11,200,000,000 CFU/ml, you can terminates fermentation, as atrophy fermentation of bacillus liquid；
Wherein, the nutrient broth medium：Peptone 10g, beef extract 3g, sodium chloride 5g, water 1000mL, pH7.2, Gu
Agar 2% is added in body culture medium；Wherein, the atrophy bacillus liquid culture medium：Glucose 15g/L, the g/ of beef extract 10
L, the g/L of peptone 5, the g/L of fish meal 5, the g/L of magnesium sulfate 0.5, the g/L of manganese sulfate 0.5, the g/L of potassium dihydrogen phosphate 0.5, phosphoric acid hydrogen
The g/L of dipotassium 0.5, calcium chloride 0.5 g/L, pH7.2；
Wherein, described fermentation medium：Glucose 15g/L corn flour 10 g/L, the g/L of corn steep liquor 30, peptone 10
G/L, the g/L of magnesium sulfate 0.5, the g/L of manganese sulfate 0.5, the g/L of potassium dihydrogen phosphate 0.5, the g/L of dipotassium hydrogen phosphate 0.5, calcium chloride 0.5
G/L, calcium carbonate 5g/L, pH7.2；
The condition of each medium sterilization is：0.10-0.15MPa, 121 DEG C sterilize 30 minutes；
2nd, by 60 parts of atrophy fermentation of bacillus liquid；Filler：28 parts of kaolin；Uv-protector：1.8 parts of trehalose, carboxylic
0.8 part of methylcellulose；Drying protectant：4 parts of sucrose, 0.8 part of sodium glutamate；It is spray-dried after mixing, obtains atrophy gemma
Bacillus pulvis；
3rd, by wetting agent：3 parts of neopelex；Dispersant：1 part of Morwet D-425；Gemma germinant：
0.8 part of fructose, 0.4 part of threonine, 2,0.5 part of dipicolimic acid 2 calcium；It is uniformly mixed so as to obtain the auxiliary agent of atrophy bacillus；
4th, the atrophy bacillus pulvis obtained by step 2 and step 3 and auxiliary agent are mixed and crossed and pass through air-flow crushing
Machine crushed 400 mesh sieves, and it is 12,400,000,000 cfu/g to detect its spore content, suspensibility 91%, wetting time 62s, moisture
For 3.55%, vacuum packaging, as atrophy bacillus wettable powder.
Embodiment 2
Application effect of the atrophy bacillus wettable powder on cucumber containing gemma germinant.
1. test site：Hunan Province Longhui County Tao Hong towns peasant household vegetables crop field.
2. test method：Divide 9th area, per the square meter of area 67, do three repetitions
Test group：The atrophy bacillus containing gemma germinant for preparing seedling with 200 times of embodiments 1 during transplanting is wettable
Property Dilution for powder liquid is stained with root 20 and divides kind, after field planting after 10 days, with the atrophy bacillus wettable powder containing gemma germinant
1000 times dilute liquid irrigating root 1 time, during cucumber growth, foliage-spray 1：500 times of wettable powders, 20 days 1 time, even spray 3 times；
Control group one：It is during transplanting that seedling is dilute with 200 times of atrophy bacillus wettable powders not containing gemma germinant
Release liquid and be stained with root 20 and divide kind, after field planting after 10 days, with 1000 times of the atrophy bacillus wettable powder not containing gemma germinant
Dilute liquid irrigating root 1 time, during cucumber growth, foliage-spray 1：500 times of atrophy bacillus not containing gemma germinant are wettable
Property pulvis, 20 days 1 time, even spray 3 times；
Control group two：With fresh water spraying and it is stained with root.
3. result of the test：After transplanting, it is found that test group is stained with the plant leaf ratio of root atrophy bacillus with control group one
Comparatively fresh, and growth potential is good, does not beat listless, is jetted through the plant of the atrophy bacillus wettable powder containing gemma germinant
Strain（Test group）Than clear water group（Control group two）6 days in advance existing flowers；The atrophy bacillus being jetted through not containing gemma germinant is wettable
The plant of property pulvis（Control group one）Than clear water group（Control group two）3 days in advance existing flowers；It is most all opened spend after observe
It was found that the bright-coloured more of the atrophy bacillus wettable powder ratio that the flowers are in blossom spray clear water are found, and the quality of flower is fine, color
Gorgeous, Hua great, flower, which looks, just to be compared " strong ", and from the point of view of the growth of whole plant, growth potential, which is significantly better than, is not jetted through gemma
Bacillus, during which used cucumber shows as that leaf color jade green, plant be neat and soil shows as loose, and cucumber is clear from mouthfeel
It is crisp, fragrant and sweet.And control group two（Clear water group）Then leaf color is yellowish green, plant is uneven.During actual survey production, add bud as can be seen from Table 1
The atrophy bacillus wettable powder of spore germinant（Test group）Compare clear water（Control group two）Volume increase 35.8%, not plus gemma
The atrophy bacillus wettable powder of germinant（Control group one）Compare clear water（Control group two）Volume increase about 14.6%, from withered
The preventing and treating situation of disease is seen, adds the atrophy bacillus wettable powder of gemma germinant（Test group）Prevention effect reach
86.01%, do not add the atrophy bacillus wettable powder of gemma germinant（Control group one）Prevention effect also reached 60%,
But the prevention effect or poor 26% or so of gemma germinant is relatively added.
Atrophy bacillus wettable powder application effect on cucumber of the table 1 containing gemma germinant
| Processing | Yield（kg） | Rate of growth (%) | Droop disease refers to | Preventive effect (%) |
| Test group | 1497 | 35.8 | 2.77 | 86.01 |
| Control group one | 1263 | 14.6 | 7.92 | 60 |
| Control group two | 1102 | 19.80 |
Claims (2)
1. a kind of atrophy bacillus wettable powder, it is characterised in that the atrophy bacillus wettable powder is by following
Parts by weight form：Atrophy fermentation of bacillus liquid 40-70 parts；Filler：Kaolin 20-30 parts；Wetting agent：DBSA
Sodium 1-4 parts；Dispersant：Morwet D-425 0.2-1 parts；Gemma germinant：Fructose 0.1-1 parts, threonine 0.1-1 parts, 2,
Dipicolimic acid 2 calcium 0.1-0.5 parts；Uv-protector：Trehalose 1-2 parts, carboxymethyl cellulose 0.2-1 parts；Dry-run protection
Agent：Sucrose 1-5 parts, sodium glutamate 0.2-1 parts；
The preparation method of the atrophy bacillus wettable powder is made up of suddenly the following steps：
First, the preparation of atrophy fermentation of bacillus liquid
Culture presevation pipe is taken out, flat board is drawn with nutrient broth solid medium and is recovered, 30 DEG C are cultivated 48 hours, under flat board
Picking single bacterium colony is seeded in 50 milliliters of nutrient broth mediums, and 30 DEG C of concussion and cultivates 24 hours in incubator, seed is adopted
With 5% inoculum concentration, the atrophy bacillus liquid culture medium being seeded in the big triangular flasks of 5L, liquid amount 2L, 30 DEG C of concussion trainings
16-28 hours are supported, detect its bacterial concentration, bacterium solution content is more than 3,000,000,000/milliliter, you can as atrophy bacillus seed liquor；
Atrophy bacillus seed liquor is seeded in 1000L fermentation tank, liquid amount is 600L fermentation medium, before throughput is
16 hours are that liquid-gas ratio per minute is 1：0.8, throughput is 1 thereafter：1；24 hours are 30 DEG C before cultivation temperature, open stirring, are turned
Speed is 160r/min, thereafter, temperature was improved into 1 DEG C in every 2 hours, cultivates 26-32 hours, detect its spore content more than 10,000,000,000
CFU/ml, you can terminate fermentation, as atrophy fermentation of bacillus liquid；
Wherein, the nutrient broth medium：Peptone 10g, beef extract 3g, sodium chloride 5g, water 1000mL, pH7.2, solid training
Support and agar 2% is added in base；Wherein, the atrophy bacillus liquid culture medium：Glucose 15g/L, the g/L of beef extract 10, egg
White peptone 5 g/L, the g/L of fish meal 5, the g/L of magnesium sulfate 0.5, the g/L of manganese sulfate 0.5, the g/L of potassium dihydrogen phosphate 0.5, dipotassium hydrogen phosphate
0.5 g/L, calcium chloride 0.5 g/L, pH7.2；
Wherein, described fermentation medium：Glucose 15g/L corn flour 10 g/L, the g/L of corn steep liquor 30, the g/L of peptone 10,
The g/L of magnesium sulfate 0.5, the g/L of manganese sulfate 0.5, the g/L of potassium dihydrogen phosphate 0.5, the g/L of dipotassium hydrogen phosphate 0.5, the g/ of calcium chloride 0.5
L, calcium carbonate 5g/L, pH7.2；
The condition of each medium sterilization is：0.10-0.15MPa, 121 DEG C sterilize 30 minutes；
2nd, by atrophy fermentation of bacillus liquid 40-70 parts；Filler：Kaolin 20-30 parts；Uv-protector：Trehalose 1-2 parts,
Carboxymethyl cellulose 0.2-1 parts；Drying protectant：Sucrose 1-5 parts, sodium glutamate 0.2-1 parts；It is spray-dried, obtains after mixing
Atrophy bacillus pulvis；
3rd, by wetting agent：Neopelex 1-4 parts；Dispersant：Morwet D-425 0.2-1 parts；Gemma is sprouted
Agent：Fructose 0.1-1 parts, threonine 0.1-1 parts, 2, dipicolimic acid 2 calcium 0.1-0.5 parts；It is uniformly mixed so as to obtain atrophy bacillus
Auxiliary agent；
4th, the atrophy bacillus pulvis obtained by step 2 and step 3 and auxiliary agent are mixed and crossed and pass through airslide disintegrating mill powder
The broken mesh sieve of mistake 400, detecting its spore content and be not less than 10,000,000,000 cfu/g, suspensibility is not less than 82%, and wetting time is not higher than 75s,
Moisture is less than 5%, vacuum packaging, as atrophy bacillus wettable powder；
The atrophy Bacillus is purchased from Chinese industrial Microbiological Culture Collection administrative center, numbering CICC
23950 。
2. a kind of atrophy bacillus wettable powder according to claim 1, its feature, is the atrophy gemma bar
Bacterium wettable powder is made up of following parts by weight：60 parts of atrophy fermentation of bacillus liquid；Filler：28 parts of kaolin；Wetting agent：Ten
3 parts of dialkyl benzene sulfonic acids sodium；Dispersant：1 part of Morwet D-425；Gemma germinant：0.8 part of fructose, 0.4 part of threonine,
2,0.5 part of dipicolimic acid 2 calcium；Uv-protector：1.8 parts of trehalose, 0.8 part of carboxymethyl cellulose；Drying protectant：Sugarcane
4 parts of sugar, 0.8 part of sodium glutamate.
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| CN102154178A (en) * | 2011-01-28 | 2011-08-17 | 广东省农业科学院植物保护研究所 | Bacillus brevis for preventing and treating hot pepper epidemic disease as well as preparation method and application of biological agent |
| CN103160450A (en) * | 2011-12-15 | 2013-06-19 | 河北农业大学 | Broad-spectrum and highly effective new Bacillus thuringiensis strain, its bacterial agent and application |
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