CN108220199A - A kind of preparation method of complex microbial inoculum - Google Patents
A kind of preparation method of complex microbial inoculum Download PDFInfo
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Abstract
The invention discloses a kind of preparation method of complex microbial inoculum, the specific steps are：Bacterial activity culture is obtained into activated spawn；The strain activated, which is inoculated into respectively in fluid nutrient medium, cultivates to obtain seed liquor, seed liquor is inoculated in cultivated in liquid medium after, concentration, dry conidia powder and fungal spore powder；It is uniformly mixed after first microbial inoculum carrier is sterilized with conidia powder, fungal spore powder, sterile Aquaponic is added in, adds sodium alginate and polyvinyl alcohol, rear constant speed is stirred evenly and is added drop-wise in sterile calcium chloride and sterile boric acid mixed solution, complex microbial inoculum is lyophilized to obtain in crosslinking curing.It has the beneficial effect that：The preparation method production cost of complex microbial inoculum of the present invention is low, the adsorbance and adsorption capacity of absorption carrier can be improved, each microorganism being capable of mutual gain in complex microbial inoculum obtained, there is efficient, Broadspectrum pest control, promote plant growth, improve crop yield and quality.
Description
Technical field
The present invention relates to agricultural biological technical fields, are especially for a kind of preparation side of complex microbial inoculum
Method.
Background technology
Biocontrol of plant disease be exactly using beneficial organism antagonism, destroy pathogen and（Or）Promoted using beneficial biology
Disease resistance is generated into plant growth, induction plant, so as to reach a measure of control disease purpose.In other words, pass through tune
The microorganism growing environment of host plant is saved, the purpose of diseases prevention disease-control is realized by the regulation and control of Tiny ecosystem effect.Plant disease
The generalized concept of biological control refers to the live body and its metabolite using beneficial organism, and the narrow sense of biocontrol of plant disease is general
The research contents for then referring to the live plant biological control of diseases using beneficial organism read：The microorganism that selecting has antagonism presses down
Pathogenic microorganism processed；Inhibit pathogenic microorganism using a series of natural antagonistic substances of microorganism, animals and plants etc. itself generation；
Disease resistance is generated using microorganism or antagonistic substance induction plant body and achievees the purpose that prevent pathogen.Most important one anti-
The biocontrol bacteria type for controlling plant disease is bacterium, and no matter biocontrol strains existing for nature itself or people use
The biocontrol strains that technological means screening obtains, biocontrol microorganisms be mainly characterized by class see fast, the inverse property of more, reproduction speed by force, to life
State is environmental-friendly, colonization ability is strong, generates antibacterial substance, promotes plant growth.It is micro- using more plant disease biological and ecological methods to prevent plant disease, pests, and erosion at present
Biology mainly has bacillus, pseudomonas, soil Pseudomonas and some other bacterium.The mechanism of biological control is profit
With between microbial species or in kind antibiosis, competition, hyperparasite, bacteriolysis or induced by microbial secondary metabolite
Plant generates disease resistance etc., to inhibit the survival of certain pathogens and activity.It is it is a kind of to environmental ecology close friend, it is sustainable
Field Plants pest control method more and more obtains various countries and plants recognizing and paying attention to for sick worker, obtained significant progress.
The prior art such as Authorization Notice No. is the Chinese invention patent of CN103651582 B, and it is compound to disclose a kind of wide spectrum
Microbial bactericide and preparation method thereof, it is by following 3 kinds of microorganism groups into former composite spore powder：Bacillus thuringiensis
(Bacillus thuringiensis) GSICC 32807, bacillus amyloliquefaciens (Bacillus amyloliquefaciens) GSICC 32826 and pythium oligandrum (Pythiumoligandrum)GSICC 61615.By two kinds
Bacterial spore powder and a kind of fungal spore powder are according to 1:1:1 weight ratio mixing, so as to which broad-spectrum composite microbial fungicide be made
Product.The broad-spectrum composite microbial fungicide product can be colonized rapidly plant rhizosphere, by with pathogen competition infect site,
The effects that hyperparasitism, secretion auxin and promotion plant nutrient absorb, significantly prevention, antagonism simultaneously kill the micro- life of various crop
Object disease；Have effects that promote long raw crop, raising yield and improving quality simultaneously.But the microbial bactericide uses the longevity
It orders short.
Invention content
The purpose of the present invention is to provide a kind of preparation method of complex microbial inoculum, the preparation method production costs
Low, low energy consumption, can improve the adsorbance and adsorption capacity of absorption carrier, each microbe species in complex microbial inoculum obtained
Collocation is reasonable, can mutual gain, have the function of efficiently, Broadspectrum pest control, promote plant growth, raising crop yield
And quality.
The problem of present invention in background technology for mentioning, the technical solution taken is：
A kind of preparation method of complex microbial inoculum, including actication of culture, conidia powder prepares, is the preparation of fungal spore powder, micro-
It is prepared by biological compound fungi agent, the specific steps are：
Actication of culture：Strain is inoculated on solid medium respectively, the active culture 20-30h under conditions of 25-30 DEG C, i.e.,
Activated spawn is obtained, it is spare, since strain is preserved in the relatively low strain library of temperature for a long time, to live before use
Change；
It is prepared by conidia powder：It is 5-10% by the bacillus amyloliquefaciens activated, bacillus polymyxa, withered grass gemma by inoculum concentration
Bacillus is inoculated into fluid nutrient medium respectively, the shaking table culture under conditions of temperature is 25-30 DEG C, rotating speed is 150-200r/min
20-30h obtains seed liquor, and then seed liquor is inoculated in liquid medium for 1.8-2.4% by inoculum concentration, is then in temperature
25-30 DEG C, rotating speed be shaking table culture 2-3d under conditions of 150-200r/min, obtain liquid bacterial suspension, three kinds of bacteriums are hanged
Supernatant liquid is prepared into conidia powder after concentrating low-temperature is dried, containing living bacteria count >=10,000,000,000/g；
The preparation of fungal spore powder：The pythium oligandrum activated is inoculated into fluid nutrient medium for 5-10% by inoculum concentration, in temperature
Spend for 25-30 DEG C, rotating speed be 150-200r/min under conditions of shaking table culture 20-30h obtain seed liquor, be then by inoculum concentration
Seed liquor is inoculated in liquid medium by 1.8-2.4%, then in the item that temperature is 25-30 DEG C, rotating speed is 150-200r/min
Shaking table culture 2-3d under part obtains liquid bacterial suspension, and three kinds of bacterial suspensions prepare fungi after concentrating low-temperature is dried
Into conidia powder, containing living bacteria count >=10,000,000,000/g；
It is prepared by complex microbial inoculum：First microbial inoculum carrier is sterilized, then is uniformly mixed with conidia powder, fungal spore powder, Ran Houzai
Sterile water, and the shaken cultivation 40-60min under the gnotobasis that rotating speed is 90-130r/min, temperature is 25-30 DEG C are added in, then
Sodium alginate and polyvinyl alcohol are added in, magnetic agitation is uniform under 120-180r/min obtains mixed liquor, by mixed liquor in 30-
Constant speed is added drop-wise in sterile calcium chloride and sterile boric acid mixed solution in 40min, the crosslinking curing under 30-35 DEG C of temperature condition
15-25h obtains gel ball, at -4 to 0 DEG C freeze-drying to moisture≤5% to get complex microbial inoculum, it is spare, on
It is 0.4-0.5 times of microbial bacterial agent weight to state sodium alginate, 2-3 times for microbial bacterial agent weight of polyvinyl alcohol, above-mentioned nothing
Contain 0-0.05mM perfluoro caprylic acids and 0-0.02mM 1,12- dodecanediol, the perfluoro caprylic acid and 1,12- dodecane in bacterium water
The addition of glycol can make starch chain length shorten and occur to be destroyed, be broken, degrade, and change the chain length of micropore starch, improve
The specific surface area and pore volume of micropore starch, and then improve it and adsorb the performance of microbial inoculum, while enable to micropore starch and bacterium
There are certain chemical bond power between agent, and then adsorption capacity of the micropore starch to microbial inoculum is reinforced, improve the sustained release of micropore starch
Performance, the final service life for improving complex microbial inoculum by sodium alginate and the crosslinking curing of chitosan, form capsule
The basic porous structure of carrier, the microbial survival time that can not only extend complex microbial inoculum is long, and product is stablized, and
Carbon source progress that extraneous microorganism under the composite bacteria agent and open condition that embed in film discharge using membrane body enable centainly
Proliferation and maintenance normal physiological biochemical activity so that the microorganism in capsule is proliferated after discharging in environment.
Preferably, conidia powder and the weight ratio of fungal spore powder are 1:0.4-0.6, bacillus amyloliquefaciens in conidia powder,
Bacillus polymyxa, bacillus subtilis weight ratio be 1:0.7-1.1:0.3-0.6, each micro- life in the complex microbial inoculum
Species collocation is reasonable, can mutual gain, lepidopterous insect pest can not only be prevented, reduce pest and sting to bite providing for pathogen invasion
" convenient channel ", and can prevent mould etc. by pythium spp, Rhizoctonia solani Kuhn, sickle-like bacteria, the pathogen of Botrytis cinerea, bread mold and column spore
Plant disease caused by pathogen can also secrete high-temperature resistant antibacterial substance, protease and auxin etc. in addition, have efficiently, extensively
The effect of the prevention and control of plant diseases, pest control is composed, while there is promoting root growth vernalization effect, plant growth can be promoted, improve crop yield and quality.
Preferably, microbial inoculum carrier and the weight ratio of microbial inoculum are 1:4-5, microbial inoculum carrier are micropore starch or bentonite or Hai Pao
Mountain flour or bamboo charcoal powder or zeolite powder, more preferably micropore starch, the microbial inoculum carrier bacterium absorption property is good, is conducive to obtain preferable
Complex microbial inoculum capsule hardness, cohesiveness and elastic recovery, the envelop rate of complex microbial inoculum, starch can be improved
Adsorption capacity can not rapidly release make complex microbial inoculum even if in swelling, it is preferably slow so as to reach
Release effect.
Preferably, the concentration of sterile calcium chloride solution and sterile boric acid solution is 3-5%, volume ratio 1:0.67-
0.82, the cross-linking reaction of α-L- guluronic acids and calcium chloride on sodium alginate is rapid, is conducive to the molding of encystation wall material, and easily
The higher gel of hardness is formed, polyvinyl alcohol can form firm chemical bond with acid reaction, and stability is good in water environment, together
When bamboo charcoal or micropore starch on-OH take part in coordination poly-merization between polyvinyl alcohol and boric acid so that polyvinyl alcohol chain obtains
Support force is obtained to be fixed on bamboo charcoal or micropore starch particle surface, increases capsule hardness.
Preferably, the application method of complex microbial inoculum is：Ditch spread spreads manuer in holes, pesticide-clay mixture, seed soaking, root dipping, sprays, pouring root.
The complex microbial inoculum can be colonized rapidly in plant rhizosphere to be grown and is formed " protective cover ", to prevent invading for root disease fungus
Dye, while with generating the substances such as antibacterial material, protease and auxin during pathogen interaction and hyperparasite, it can be significantly pre-
Anti-, antagonism directly kills crops multiple diseases, using the complex microbial inoculum can to avoid with reduction chemical bactericide
Dosage, reduce pollution of the chemical agent to environment, reduce the drug resistance of pathogen, and then reduce the input cost of disease, warp
Ji environmental protection, the complex microbial inoculum service life is long, and in root, use can continue 3 months or more, and being sprayed in leaf portion can be with
Continue 7-15 days, reduce times for spraying and working feature, lasting period are long.
Compared with prior art, the advantage of the invention is that：1）The preparation method production of complex microbial inoculum of the present invention
At low cost, low energy consumption, easily operated, is easy to industrialized production, has larger development potentiality；2）The preparation method can improve
The adsorbance and adsorption capacity of absorption carrier, improve the sustained release performance of complex microbial inoculum, finally improve complex microbial inoculum
Service life, and can so that capsule in microorganism be proliferated after discharging in environment；3）The complex microbial inoculum
In the collocation of each microbe species it is reasonable, can mutual gain, there is efficient, Broadspectrum pest control, while there is rush
Root vernalization acts on, and can promote plant growth, improve crop yield and quality；4）Using the complex microbial inoculum can to avoid with
The dosage of chemical bactericide is reduced, reduces pollution of the chemical agent to environment, reduces the drug resistance of pathogen, and then reduces disease
Input cost, it is economic and environment-friendly.
Specific embodiment
Below by embodiment, the invention will be further described：
Embodiment 1：
A kind of preparation method of complex microbial inoculum, including actication of culture, conidia powder prepares, is the preparation of fungal spore powder, micro-
It is prepared by biological compound fungi agent, the specific steps are：
1）Actication of culture：Strain is inoculated on solid medium respectively, under conditions of 30 DEG C active culture 20h to get to
Activated spawn, it is spare, since strain is preserved in the relatively low strain library of temperature for a long time, to be activated before use；
2）It is prepared by conidia powder：By inoculum concentration for 10% by the bacillus amyloliquefaciens activated, bacillus polymyxa, withered grass gemma
Bacillus is inoculated into fluid nutrient medium respectively, and shaking table culture 20h must be planted under conditions of temperature is 25 DEG C, rotating speed is 200r/min
Then seed liquor is inoculated in liquid medium by inoculum concentration for 2.4% by sub- liquid, then temperature is 25 DEG C, rotating speed is
Shaking table culture 2d under conditions of 200r/min, obtains liquid bacterial suspension, and three kinds of bacterial suspensions are dried by concentrating low-temperature
Afterwards, conidia powder is prepared into, containing living bacteria count >=10,000,000,000/g；
3）The preparation of fungal spore powder：The pythium oligandrum activated is inoculated into fluid nutrient medium for 10% by inoculum concentration, in temperature
Spend for 25 DEG C, rotating speed be 200r/min under conditions of shaking table culture 20h obtain seed liquor, then by inoculum concentration for 2.4% by seed liquor
It is inoculated in liquid medium, then the shaking table culture 2d under conditions of temperature is 25 DEG C, rotating speed is 200r/min, obtains liquid
Bacterial suspension, three kinds of bacterial suspensions prepare fungi into conidia powder after concentrating low-temperature is dried, containing living bacteria count >=
10000000000/g；
4）It is prepared by complex microbial inoculum：First microbial inoculum carrier is sterilized, then is uniformly mixed with conidia powder, fungal spore powder, then
Sterile water, and the shaken cultivation 60min under the gnotobasis that rotating speed is 130r/min, temperature is 25 DEG C are added, adds sea
Mosanom and polyvinyl alcohol, magnetic agitation is uniform under 120r/min obtains mixed liquor, and by mixed liquor, constant speed is added drop-wise in 40min
In sterile calcium chloride and sterile boric acid mixed solution, crosslinking curing 25h obtains gel ball under 30 DEG C of temperature condition, then at -4
Freeze-drying is to moisture≤5% to get complex microbial inoculum at DEG C, and spare, above-mentioned sodium alginate is microbial bacterial agent weight
0.5 times, polyvinyl alcohol is 2 times of microbial bacterial agent weight, passes through sodium alginate and the crosslinking curing of chitosan, forms capsule
The basic porous structure of carrier, the microbial survival time that can not only extend complex microbial inoculum is long, and product is stablized, and
Carbon source progress that extraneous microorganism under the composite bacteria agent and open condition that embed in film discharge using membrane body enable centainly
Proliferation and maintenance normal physiological biochemical activity so that the microorganism in capsule is proliferated after discharging in environment.
Above-mentioned conidia powder and the weight ratio of fungal spore powder are 1:0.6, bacillus amyloliquefaciens, mostly viscous gemma in conidia powder
Bacillus, bacillus subtilis weight ratio be 1:0.7:0.6, each microbe species collocation is reasonable in the complex microbial inoculum,
Can mutual gain, can not only prevent lepidopterous insect pest, reduce pest and " the convenient channel " bitten and provided for pathogen invasion is provided, and
And the plant as caused by pythium spp, Rhizoctonia solani Kuhn, sickle-like bacteria, the pathogen of Botrytis cinerea, bread mold and the pathogens such as column spore is mould can be prevented
Object disease can also secrete high-temperature resistant antibacterial substance, protease and auxin etc. in addition, have efficient, Broadspectrum pest control
Effect, while there is promoting root growth vernalization effect, plant growth can be promoted, improve crop yield and quality.
Above-mentioned microbial inoculum carrier and the weight ratio of microbial inoculum are 1:4, microbial inoculum carrier is micropore starch or bentonite or sepiolite powder
Or bamboo charcoal powder or zeolite powder, more preferably micropore starch, the microbial inoculum carrier bacterium absorption property is good, is conducive to obtain preferably micro-
Biological compound fungi agent capsules hardness, cohesiveness and elastic recovery can improve the envelop rate of complex microbial inoculum, the suction of starch
Attached ability can not be released rapidly making complex microbial inoculum even if in swelling, so as to reach better sustained release effect
Fruit.
The concentration of above-mentioned sterile calcium chloride solution and sterile boric acid solution is 5%, volume ratio 1:0.67, alginic acid
The cross-linking reaction of α-L- guluronic acids and calcium chloride on sodium is rapid, be conducive to encystation wall material molding, and easily formed hardness compared with
High gel, polyvinyl alcohol can form firm chemical bond with acid reaction, and stability is good in water environment, while bamboo charcoal or micro-
- OH on the starch of hole takes part in the coordination poly-merization between polyvinyl alcohol and boric acid so that polyvinyl alcohol chain is able to be fixed on bamboo
Charcoal or micropore starch particle surface increase capsule hardness so as to obtain support force.
The application method of mentioned microorganism composite bacteria agent is：Ditch spread spreads manuer in holes, pesticide-clay mixture, seed soaking, root dipping, sprays, pouring root.It should
Complex microbial inoculum can be colonized rapidly in plant rhizosphere to be grown and is formed " protective cover ", to prevent invading for root disease fungus
Dye, while with generating the substances such as antibacterial material, protease and auxin during pathogen interaction and hyperparasite, it can be significantly pre-
Anti-, antagonism directly kills crops multiple diseases, using the complex microbial inoculum can to avoid with reduction chemical bactericide
Dosage, reduce pollution of the chemical agent to environment, reduce the drug resistance of pathogen, and then reduce the input cost of disease, warp
Ji environmental protection, the complex microbial inoculum service life is long, and in root, use can continue 3 months or more, and being sprayed in leaf portion can be with
Continue 7-15 days, reduce times for spraying and working feature, lasting period are long.
Embodiment 2：
A kind of preparation method of complex microbial inoculum, including actication of culture, conidia powder prepares, is the preparation of fungal spore powder, micro-
It is prepared by biological compound fungi agent, the specific steps are：
1）Actication of culture：Strain is inoculated on solid medium respectively, under conditions of 25 DEG C active culture 30h to get to
Activated spawn, it is spare；
2）It is prepared by conidia powder：By inoculum concentration for 5% by the bacillus amyloliquefaciens activated, bacillus polymyxa, bacillus subtilis
Bacterium is inoculated into fluid nutrient medium respectively, and shaking table culture 30h obtains seed under conditions of temperature is 30 DEG C, rotating speed is 150r/min
Then seed liquor is inoculated in liquid medium by inoculum concentration for 1.8% by liquid, then temperature be 30 DEG C, rotating speed 150r/
Shaking table culture 3d under conditions of min obtains liquid bacterial suspension, and three kinds of bacterial suspensions are after concentrating low-temperature is dried, system
It is standby into conidia powder, containing living bacteria count >=10,000,000,000/g；
3）The preparation of fungal spore powder：The pythium oligandrum activated is inoculated into fluid nutrient medium for 5% by inoculum concentration, in temperature
Spend for 30 DEG C, rotating speed be 150r/min under conditions of shaking table culture 30h obtain seed liquor, then by inoculum concentration for 1.8% by seed liquor
It is inoculated in liquid medium, then the shaking table culture 3d under conditions of temperature is 30 DEG C, rotating speed is 150r/min, obtains liquid
Bacterial suspension, three kinds of bacterial suspensions prepare fungi into conidia powder after concentrating low-temperature is dried, containing living bacteria count >=
10000000000/g；
4）It is prepared by complex microbial inoculum：First microbial inoculum carrier is sterilized, then is uniformly mixed with conidia powder, fungal spore powder, then
Sterile water, and the shaken cultivation 40min under the gnotobasis that rotating speed is 90r/min, temperature is 30 DEG C are added, adds seaweed
Sour sodium and polyvinyl alcohol, magnetic agitation is uniform under 180r/min obtains mixed liquor, and by mixed liquor, constant speed is added drop-wise to nothing in 30min
In bacterium calcium chloride and sterile boric acid mixed solution, crosslinking curing 15h obtains gel ball under 35 DEG C of temperature condition, then at 0 DEG C
Lower freeze-drying is to moisture≤5% to get complex microbial inoculum, and spare, above-mentioned sodium alginate is microbial bacterial agent weight
0.4 times, polyvinyl alcohol is 3 times of microbial bacterial agent weight, and 0.02mM 1,12- dodecanediols are contained in sterile water；Above-mentioned spore
The weight ratio of sub- powder and fungal spore powder is 1:0.4, bacillus amyloliquefaciens, bacillus polymyxa, withered grass gemma in conidia powder
The weight ratio of bacillus is 1:1.1:0.3；Microbial inoculum carrier and the weight ratio of microbial inoculum are 1:5, microbial inoculum carrier is micropore starch；Sterile chlorine
The concentration for changing calcium solution and sterile boric acid solution is 5%, volume ratio 1:0.67.
The application method of mentioned microorganism composite bacteria agent is：Ditch spread.
Embodiment 3：
A kind of preparation method of complex microbial inoculum, including actication of culture, conidia powder prepares, is the preparation of fungal spore powder, micro-
It is prepared by biological compound fungi agent, the specific steps are：
1）Actication of culture：Strain is inoculated on solid medium respectively, under conditions of 28 DEG C active culture 25h to get to
Activated spawn, it is spare；
2）It is prepared by conidia powder：By inoculum concentration for 8% by the bacillus amyloliquefaciens activated, bacillus polymyxa, bacillus subtilis
Bacterium is inoculated into fluid nutrient medium respectively, and shaking table culture 25h obtains seed under conditions of temperature is 28 DEG C, rotating speed is 180r/min
Then seed liquor is inoculated in liquid medium by inoculum concentration for 2.0% by liquid, then temperature be 28 DEG C, rotating speed 180r/
Shaking table culture 2.5d under conditions of min obtains liquid bacterial suspension, three kinds of bacterial suspensions after concentrating low-temperature is dried,
Conidia powder is prepared into, containing living bacteria count >=10,000,000,000/g；
3）The preparation of fungal spore powder：The pythium oligandrum activated is inoculated into fluid nutrient medium for 8% by inoculum concentration, in temperature
Spend for 28 DEG C, rotating speed be 180r/min under conditions of shaking table culture 25h obtain seed liquor, then by inoculum concentration for 2.0% by seed liquor
It is inoculated in liquid medium, then the shaking table culture 2.5d under conditions of temperature is 28 DEG C, rotating speed is 180r/min, obtains liquid
Body bacterial suspension, three kinds of bacterial suspensions prepare fungi into conidia powder after concentrating low-temperature is dried, containing living bacteria count >=
10000000000/g；
4）It is prepared by complex microbial inoculum：First microbial inoculum carrier is sterilized, then is uniformly mixed with conidia powder, fungal spore powder, then
Sterile water, and the shaken cultivation 50min under the gnotobasis that rotating speed is 110r/min, temperature is 28 DEG C are added, adds sea
Mosanom and polyvinyl alcohol, magnetic agitation is uniform under 150r/min obtains mixed liquor, and by mixed liquor, constant speed is added drop-wise in 35min
In sterile calcium chloride and sterile boric acid mixed solution, crosslinking curing 20h obtains gel ball under 32 DEG C of temperature condition, then at -2
Freeze-drying is to moisture≤5% to get complex microbial inoculum at DEG C, and spare, above-mentioned sodium alginate is microbial bacterial agent weight
0.45 times, polyvinyl alcohol is 2.5 times of microbial bacterial agent weight, and 0.02mM perfluoro caprylic acids and 0.01mM are contained in sterile water
1,12- dodecanediols；Above-mentioned conidia powder and the weight ratio of fungal spore powder are 1:0.5, bacillus amyloliquefaciens in conidia powder,
Bacillus polymyxa, bacillus subtilis weight ratio be 1:0.9:0.5；Microbial inoculum carrier and the weight ratio of microbial inoculum are 1:4.5, bacterium
Agent carrier is micropore starch；The concentration of sterile calcium chloride solution and sterile boric acid solution is 4%, volume ratio 1:0.75.
The application method of mentioned microorganism composite bacteria agent is：Seed soaking.
Routine operation in the operating procedure of the present invention is well known to those skilled in the art, herein without repeating.
Technical scheme of the present invention is described in detail in embodiment described above, it should be understood that the above is only
For specific embodiments of the present invention, it is not intended to restrict the invention, all any modifications made in the spirit of the present invention,
Supplement or similar fashion replacement etc., should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of preparation method of complex microbial inoculum, including actication of culture, conidia powder prepare, the preparation of fungal spore powder,
It is prepared by complex microbial inoculum, it is characterised in that：The complex microbial inoculum preparation process is：First microbial inoculum carrier is sterilized,
It is uniformly mixed again with conidia powder, fungal spore powder, then adds sterile water, shaken cultivation adds sodium alginate and poly- second
Enol, magnetic agitation is uniform to obtain mixed liquor, and mixed liquor constant speed is added drop-wise in sterile calcium chloride and sterile boric acid mixed solution, is handed over
Connection curing obtains gel ball, is lyophilized to get complex microbial inoculum.
2. a kind of preparation method of complex microbial inoculum according to claim 1, it is characterised in that：The conidia powder and
The weight ratio of fungal spore powder is 1:0.4-0.6, bacillus amyloliquefaciens, bacillus polymyxa, bacillus subtilis in conidia powder
The weight ratio of bacterium is 1:0.7-1.1:0.3-0.6.
3. a kind of preparation method of complex microbial inoculum according to claim 1, it is characterised in that：The microbial inoculum carrier
Weight ratio with microbial inoculum is 1:4-5, microbial inoculum carrier are micropore starch or bentonite or sepiolite powder or bamboo charcoal powder or zeolite powder.
4. a kind of preparation method of complex microbial inoculum according to claim 1, it is characterised in that：The shaken cultivation
Rotating speed be 90-130r/min, temperature is 25-30 DEG C, time 40-60min.
5. a kind of preparation method of complex microbial inoculum according to claim 1, it is characterised in that：In the sterile water
Contain 0-0.05mM perfluoro caprylic acids and 0-0.02mM 1,12- dodecanediols.
6. a kind of preparation method of complex microbial inoculum according to claim 1, it is characterised in that：The sodium alginate
It it is 0.4-0.5 times of microbial bacterial agent weight, polyvinyl alcohol is 2-3 times of microbial bacterial agent weight.
7. a kind of preparation method of complex microbial inoculum according to claim 1, it is characterised in that：The sterile chlorination
The concentration of calcium solution and sterile boric acid solution is 3-5%, volume ratio 1:0.67-0.82, the crosslinking curing temperature are
30-35 DEG C, time 15-25h.
8. a kind of preparation method of complex microbial inoculum according to claim 1, it is characterised in that：The conidia powder system
It is for step：The bacillus amyloliquefaciens activated, bacillus polymyxa, bacillus subtilis are divided for 5-10% by inoculum concentration
It is not inoculated into fluid nutrient medium, the shaking table culture 20-30h under conditions of temperature is 25-30 DEG C, rotating speed is 150-200r/min
Seed liquor is obtained, then seed liquor is inoculated in liquid medium for 1.8-2.4% by inoculum concentration, is then 25-30 in temperature
DEG C, rotating speed be shaking table culture 2-3d under conditions of 150-200r/min, obtain liquid bacterial suspension, three kinds of bacterial suspensions warps
After crossing concentration low temperature drying, conidia powder is prepared into, containing living bacteria count >=10,000,000,000/g.
9. a kind of preparation method of complex microbial inoculum according to claim 1, it is characterised in that：The fungal spore
The preparation process of powder is：The pythium oligandrum activated is inoculated into fluid nutrient medium for 5-10% by inoculum concentration, is in temperature
25-30 DEG C, rotating speed obtains seed liquor for shaking table culture 20-30h under conditions of 150-200r/min, be then 1.8- by inoculum concentration
2.4% is inoculated in seed liquor in liquid medium, then under conditions of temperature is 25-30 DEG C, rotating speed is 150-200r/min
Shaking table culture 2-3d obtains liquid bacterial suspension, and three kinds of bacterial suspensions prepare fungi into spore after concentrating low-temperature is dried
Sub- powder, containing living bacteria count >=10,000,000,000/g.
10. according to a kind of preparation method of complex microbial inoculum of claim 1 to 9 any one of them, it is characterised in that：Institute
The application method for stating complex microbial inoculum is：Ditch spread spreads manuer in holes, pesticide-clay mixture, seed soaking, root dipping, sprays, pouring root.
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| CN108904849A (en) * | 2018-09-07 | 2018-11-30 | 浙江九田环保科技有限公司 | Microbial deodorant and its manufacturing method |
| CN109042638A (en) * | 2018-08-08 | 2018-12-21 | 长沙昱旻信息科技有限公司 | A kind of controlled release bacillus subtilis microbial agent |
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