CN109082391A - A kind of controlled release agricultural microbial inoculum - Google Patents

A kind of controlled release agricultural microbial inoculum Download PDF

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Publication number
CN109082391A
CN109082391A CN201810899582.XA CN201810899582A CN109082391A CN 109082391 A CN109082391 A CN 109082391A CN 201810899582 A CN201810899582 A CN 201810899582A CN 109082391 A CN109082391 A CN 109082391A
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microorganism
microbial inoculum
product
binder
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CN109082391B (en
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温岚
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Changsha Yuzhang Information Technology Co Ltd
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Changsha Yuzhang Information Technology Co Ltd
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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
    • 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
    • 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
    • A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
    • 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
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor

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  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Mycology (AREA)
  • Medicinal Chemistry (AREA)
  • Pest Control & Pesticides (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Environmental Sciences (AREA)
  • Agronomy & Crop Science (AREA)
  • Biomedical Technology (AREA)
  • Plant Pathology (AREA)
  • Dentistry (AREA)
  • Botany (AREA)
  • Fertilizers (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
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Abstract

The present invention provides a kind of controlled release agricultural microbial inoculum, the microbial inoculum includes microorganism, binder, dopamine or its salt, the high-polymer molecule.The present invention is by being attached to the surface that agricultural uses microbial inoculum particle for dopamine, then the high-polymer molecule of tool environmental sensitivity is coated on dopamine surface, it can be according to the variation of environmental condition to prepare one kind, and the burst size for the viable bacteria wrapped up in it is automatically adjusted, it is a kind of novel wisdom microbial inoculum that microbial inoculum utilization efficiency can be improved.Controlled release agricultural microbial inoculum of the present invention, can be released effectively viable bacteria, so as to inhibit harmful microbe movable, improve the disease-resistant anti-adversity ability of crop, the improvement soil fertility of sustainability.Controlled release agricultural microbial inoculum of the present invention, preparation process is simple and environmentally-friendly, does not use organic solvent, is suitable for industrial mass production application.

Description

A kind of controlled release agricultural microbial inoculum
Technical field
The invention belongs to the development technique fields of agriculture microbial inoculum, and in particular to a kind of controlled release agricultural microbial inoculum.
Background technique
The application of microbial bacterial agent can be such that Soil Microorganism amount dramatically increases, and the activity of these increased microorganisms Can promote the enhancing of soil enzyme activities and soil slightly solubility mineral nutrients can be made to be decomposed and be discharged, at the same time this Biology can also secrete plant hormone slightly, to promote plant growth.
The use of microbial bacterial agent greatly reduces the dosage of chemical fertilizer and reduces pollution, and can be by between microorganism Or the interaction between plant enhances the disease-resistant and anti-adversity ability of plant, this is for improving crops quality and yield, extension The picking time of agricultural product and plants important in inhibiting.Microbial bacterial agent is safe and convenient to use, and cheap not dirty Environment is contaminated, is a kind of following emerging preparation for replacing chemical fertilizer.
Summary of the invention
The present invention provides a kind of controlled release agricultural microbial inoculum, the present invention is by being attached to agricultural microbial inoculum particle for dopamine Surface, then dopamine surface coat tool environmental sensitivity the high-polymer molecule, so that preparing one kind can be according to environment The variation of condition, and the burst size for the viable bacteria wrapped up in it is automatically adjusted, it is a kind of novel intelligence that microbial inoculum utilization efficiency can be improved Intelligent microbial inoculum.
It is an object of the present invention to provide a kind of microbial inoculum, the microbial inoculum include microorganism, binder, dopamine or its Salt, the high-polymer molecule.
Specifically, the microbial inoculum further includes following 1) -4) described at least one:
1) binder includes that the nutriment of its metabolism or breeding can be provided for the microorganism；Specifically, the bonding Agent includes starch and/or water；
2) microorganism includes the microorganism beneficial to plant growth；Specifically, the microorganism includes bacillus subtilis Bacterium, saccharomycete and/or lactic acid bacteria；Again specifically, when the microorganism includes bacillus subtilis, saccharomycete and lactic acid bacteria, Bacillus subtilis, saccharomycete, lactic acid bacteria mass ratio be 1: 1: 1；Specifically, the plant includes crops；
3) the high-polymer molecule includes temperature sensitive the high-polymer molecule；Specifically, the high-polymer molecule includes PNIPAm poly- Close object；
4) dopamine or its salt are covered on the surface of binder and microorganism, and the high-polymer molecule is covered on described The surface of dopamine or its salt；Specifically, the binder includes that can provide the nutriment of its metabolism or breeding for the microorganism； Again specifically, the binder includes starch and/or water；Specifically, the microorganism includes bacillus subtilis, saccharomycete And/or lactic acid bacteria；Again specifically, when the binder includes starch and water, when the microorganism include bacillus subtilis, When saccharomycete and lactic acid bacteria, bacillus subtilis, saccharomycete, lactic acid bacteria, starch and water mass ratio be 2: 2: 2: 2: 1.
Specifically, the microbial inoculum is prepared by the following method:
After microorganism and binder mixing granulation, in the Tris-HCl buffer of pH value 8~8.5, with hydrochloric acid DOPA Amine is stirred at room temperature 2h or more together, after suction filtration, place the product in reaction vessel, adds ultrapure water, leads to N2, it is added into solution high Monomer, crosslinking agent, the catalyst of polymers molecule are protected from light and continue logical N2, stir at room temperature, reaction vessel be then transferred to ice bath Middle stirring is added initiator, is transferred in closed mold after stirring, polymerize for 24 hours at room temperature, product is crushed to obtain the final product；
Specifically, by mass, in the method, described microorganism 1-10 parts, 1-10 parts of binder, Dopamine hydrochloride 1- 10 parts, 0.8-200 parts of 10-500 parts of monomer, the crosslinking agent of the high-polymer molecule；
Again specifically, by mass, in the method, 7.5 parts of the microorganism, 2.5 parts of binder, Dopamine hydrochloride 1 Part, 1-20 parts of 12-60 parts of monomer, the crosslinking agent of the high-polymer molecule；
Specifically, the binder includes starch and/or water；Specifically, the microorganism includes bacillus subtilis, ferment Female bacterium and/or lactic acid bacteria；Again specifically, when the binder includes starch and water, when the microorganism includes withered grass gemma When bacillus, saccharomycete and lactic acid bacteria, bacillus subtilis, saccharomycete, lactic acid bacteria, starch and water mass ratio be 2: 2: 2: 2: 1.
Of the invention a further object is provides a kind of preparation method of microbial inoculum, which comprises by microorganism and glues After tying agent mixing granulation, in the Tris-HCl buffer of pH value 8~8.5,2h or more is stirred at room temperature together with Dopamine hydrochloride, After suction filtration, place the product in reaction vessel, add ultrapure water, leads to N2, monomer, the crosslinking of the high-polymer molecule are added into solution Agent, catalyst are protected from light and continue logical N2, it stirs at room temperature, then reaction vessel is transferred in ice bath and is stirred, initiator is added, It is transferred in closed mold after stirring, polymerize at room temperature for 24 hours, product is crushed to obtain the final product.
Specifically, the method also includes following 1) -6) described at least one:
1) binder includes that the nutriment of its metabolism or breeding can be provided for the microorganism；Specifically, the bonding Agent includes starch and/or water；
2) microorganism includes the microorganism beneficial to plant growth；Specifically, the microorganism includes bacillus subtilis Bacterium, saccharomycete and/or lactic acid bacteria；Again specifically, when the microorganism includes bacillus subtilis, saccharomycete and lactic acid bacteria, Bacillus subtilis, saccharomycete, lactic acid bacteria mass ratio be 1: 1: 1；Specifically, the plant includes crops；
3) the high-polymer molecule includes temperature sensitive the high-polymer molecule；Specifically, the high-polymer molecule includes PNIPAm poly- Close object；
4) by mass, in the method, described microorganism 1-10 parts, 1-10 parts of binder, 1-10 parts of Dopamine hydrochloride, 0.8-200 parts of 10-500 parts of monomer, the crosslinking agent of the high-polymer molecule；
Specifically, by mass, in the method, 7.5 parts of the microorganism, 2.5 parts of binder, 1 part of Dopamine hydrochloride, 1-20 parts of 12-60 parts of monomer, the crosslinking agent of the high-polymer molecule；
Again specifically, the binder includes starch and/or water；Specifically, the microorganism include bacillus subtilis, Saccharomycete and/or lactic acid bacteria；Again specifically, when the binder includes starch and water, when the microorganism includes withered grass bud When spore bacillus, saccharomycete and lactic acid bacteria, bacillus subtilis, saccharomycete, lactic acid bacteria, starch and water mass ratio be 2: 2: 2: 2: 1；
5) granulation includes that High Speed Stirring Machine granulation, 30-37 DEG C of fluidized drying is added in microorganism and binder To water content 3-5%, cross 20 meshes to obtain the final product；
6) more with hydrochloric acid in the Tris-HCl buffer of pH value 8~8.5 by after microorganism and binder mixing granulation Bar amine is stirred at room temperature 2h or more together, after suction filtration, place the product in reaction vessel, adds ultrapure water, leads to N2, it is added into solution Monomer NIPAM, crosslinking agent and catalyst, dosage are respectively that 50ml ultrapure water, NIPAM 11.3g/ is added in product described in every 1-5g 100g H2O, crosslinking agent clay 0.8-4g/100gH28 μ L/100gH of O, catalyst TEMED2O；It is protected from light and continues logical N2, at room temperature 15min is stirred, then reaction vessel is transferred in ice bath and stirs 5min, initiator KPS is added, KPS dosage is 2g/100g H2O is transferred in closed mold after stirring 30s, polymerize at room temperature for 24 hours.
Of the invention a further object is provides the present invention microbial inoculum that any the method is prepared.
It is also another object of the present invention to provide the applications of any microbial inoculum of the present invention.
Specifically, the application includes following 1) -4) described at least one:
1) sustained release or controlled release product as the bacterium of useful plants growth；
2) product as useful plants growth；
3) as Ecological Distribution of Soil Microorganisms or the adjusting product of nutrition condition；
4) as the sustained release of fertilizer or controlled release product.
The plant includes crops.
Specifically, the sustained release or controlled release are temperature when the high-polymer molecule in the microbial inoculum includes PNIPAm polymer Spend the sustained release or controlled release of response mechanism；Specifically, when temperature is lower than the LCST of PNIPAm polymer, the burst size of the product Increase, when temperature is higher than the LCST of PNIPAm polymer, the burst size of the product is reduced.
Final object of the present invention is to provide the application of any the method for the present invention.
Specifically, the application includes following 1) -4) described at least one:
1) application in the product or its Related product that preparation has the bacterium of sustained release or the growth of controlled release useful plants；
2) application in the product or its Related product that preparation has useful plants growth；
3) there is the application in the product or its Related product for adjusting Ecological Distribution of Soil Microorganisms or nutrition condition in preparation；
4) application in the product or its Related product that preparation has sustained release or release and release control fertilizer.
The plant includes crops.
Specifically, the sustained release or controlled release are temperature when the high-polymer molecule in the method includes PNIPAm polymer Spend the sustained release or controlled release of response mechanism；Specifically, when temperature is lower than the LCST of PNIPAm polymer, the burst size of the product Increase, when temperature is higher than the LCST of PNIPAm polymer, the burst size of the product is reduced.
One kind provided by the invention, has at least the following advantages:
(1) controlled release agricultural microbial inoculum provided by the invention, can be released effectively viable bacteria, so as to improve the nutritional condition of crop With nutrient supply situation, inhibits harmful microbe activity, improve the disease-resistant anti-adversity ability of crop, the improvement soil of sustainability Fertility state.
(2) controlled release agricultural microbial inoculum provided by the invention, preparation process is simple and environmentally-friendly, does not use organic solvent, is suitble to In industrial mass production application.
(3) in a specific embodiment, the present invention is pelletized after being mixed using starch with viable bacteria, and gained particle is water-soluble Property is poor, is able to maintain aggregate state in water, is conducive to the cladding of dopamine or high polymer；In addition, the starch in particle can be made For nutriment, be conducive to improve number of viable, viable bacteria also helps intragranular disintegration release to the degradation of starch simultaneously and lives Bacterium need not add additional disintegrating agent in pelletization.
(4) in a specific embodiment, the present invention utilizes the temperature sensitive of agricultural microbial inoculum SiC p surface plating The LCST of PNIPAm polymer is near 32 °, and when temperature is lower than LCST, PNIPAM strand has stronger hydrophily, table It is now the ball of string structure of stretching, extension, is easy release viable bacteria；When temperature rises on LCST, PNIPAM brush is in collapsing shape State, so that the agricultural of multiple layer polymer coating is reduced with microbial inoculum particulate permeability, to slow down rate of release, to reach according to ring The purpose of the variation intelligent control microbial inoculum performance in border, improves the utilization efficiency of microbial inoculum.
(5) controlled release agricultural microbial inoculum provided by the invention, in a specific embodiment, temperature sensitive PNIPAm polymer It is biodegradable, problem of environmental pollution brought by a large amount of applications of fertilizer is avoided, ecological ideas sustainable development is met It is required that having huge reality and commercial application value.
Specific embodiment
Made in following embodiments, experimental method is conventional method unless otherwise specified.
Material as used in the following examples etc., is commercially available unless otherwise specified.
Following embodiments and its illustrate for explanation and understanding the present invention, do not constitute to improper limit of the invention It is fixed.
The preparation of embodiment 1, temperature-responsive controlled release agricultural microbial inoculum
(1) the agricultural preparation of microbial inoculum granule
It is 2: 2: 2: 2: 1 bacillus subtilis by mass ratio, saccharomycete, lactic acid bacteria, starch, water addition high-speed stirring mixing Grain mechanism grain, 30-37 DEG C of fluidized drying to water content 3-5% cross 20 meshes up to agricultural microbial inoculum granule.
(2) agricultural microbial inoculum granule surface package attachment dopamine
It weighs 1.5g Tris to be placed in a beaker, 900mL deionized water about is added, with the HCl of 6M by Tris-HCl solution PH is adjusted to 8~8.5；The agricultural of above-mentioned preparation microbial inoculum granule 50g is added in above-mentioned Tris-HCl solution, and solution is determined Hold to 1L；5g Dopamine hydrochloride is added, 2h or more is stirred at room temperature, filter surface package is attached with the agricultural microbial inoculum of dopamine Particle.
(3) cladding of temperature sensitive the high-polymer molecule material
It takes the surface of the above-mentioned preparation of 1-5g to wrap up the agricultural microbial inoculum particle for being attached with dopamine, is placed in the reaction of 100ml In container, add ultrapure water 50ml.Logical N20.5h removes dissolved oxygen therein, and monomer NIPAM, crosslinking agent are added into solution and urges Agent, specific dosage are respectively NIPAM 11.3g/100g H2O, crosslinking agent clay 0.8-4g/100g H2O, catalyst TEMED 8μL/100gH2O is protected from light under tinfoil cladding and continues logical N2, 15min is quickly stirred at room temperature.Then reaction vessel is transferred to 5min is slowly stirred in ice bath, initiator KPS is added, and (KPS dosage is 2g/100g H2O), it is transferred quickly to after stirring 30s close In die closing tool, it polymerize at room temperature for 24 hours.Product centrifuge washing in ultrapure water removes unreacted monomer and impurity.Finally will Product crushes, and obtains surface and coat the agricultural of temperature sensitive the high-polymer molecule with microbial inoculum particle using bacterium to get temperature-responsive controlled release agricultural Agent.
The viable bacteria release experiment of embodiment 2, temperature-responsive controlled release agricultural microbial inoculum
The temperature-responsive controlled release agricultural microbial inoculum 18g prepared according to 1 the method for embodiment is taken, is equally divided into 9 parts, respectively 10 days, 1 month and 3 months are placed in 15 DEG C, 25 DEG C, 35 DEG C of environment, are then detected under different temperatures different standing times, The viable bacteria burst size of temperature-responsive controlled release agricultural microbial inoculum.
Plate detection method detects viable bacteria and discharges quantity measuring method:
1) it is configured to solution according to the ratio that the mass volume ratio of sample and sterile water after placement is 1: 10, it will be described molten Liquid filtered 3 micron membrane filters, with filter off do not discharge viable bacteria surface package be attached with dopamine agricultural microbial inoculum particle and Surface coats the agricultural microbial inoculum of temperature sensitive the high-polymer molecule；
2) collect filtrate, be added bead, stand 20min, on rotary shaker 200r/min vibrate 30min to get Mother liquor bacteria suspension；
3) it draws the above-mentioned mother liquor bacteria suspension of 5.0ml respectively with Sterile pipette to be added in 45ml sterile water, is by volume 1 : 10 ratio is serially diluted, and a series of dilution bacteria suspensions of different extension rates are respectively obtained；
4) each sample takes 3 dilutions for being continuously suitable for, and draws different dilution bacteria suspensions respectively with Sterile pipette 0.1ml is added on solid agar medium, and the bacteria suspension of different dilutions is uniformly applied to solid with sterile glass scraper respectively On agar medium；Each dilution is repeated 3 times, while making blank control with sterile water, is trained in 37 DEG C of constant incubator Support 12-36h；
5) bacterium colony count and sample in viable count purpose calculate: according to national standards the method in GB20287-2006 into Row.
Testing result is as shown in table 1.Table 1 the result shows that, the temperature-responsive control being prepared according to 1 the method for embodiment Releasing agriculture microbial inoculum according to the difference of temperature, can adjust the rate of release of viable bacteria of its package, when temperature is lower than 32 DEG C of LCST When, temperature is lower, and the burst size of viable bacteria is bigger；When temperature is higher than 32 DEG C of LCST, the burst size of viable bacteria sharply declines.This The experimental results showed that temperature-responsive controlled release agricultural provided by the invention is very suitable to the fertilising of early rice and winter wheat with microbial inoculum.And The summer of high temperature, the viable bacteria that temperature-responsive controlled release agricultural microbial inoculum provided by the invention can automatically save its package are not lost, mention The utilization efficiency of high microbial inoculum.
In addition, same temperature, different standing times, the burst size of viable bacteria in sample, thus it is speculated that should by bacterium metabolic activity and The consumption of nutriment is related, if the sample is placed in field, the quantity of viable bacteria should be greater than quantity shown in table 1, because Be conducive to the metabolism breeding of bacterium for field conditions.
Table 1
Embodiments of the present invention above described embodiment only expresses, the description thereof is more specific and detailed, but can not Therefore limitations on the scope of the patent of the present invention are interpreted as, as long as skill obtained in the form of equivalent substitutions or equivalent transformations Art scheme should all be fallen within the scope and spirit of the invention.

Claims (10)

1. a kind of microbial inoculum, which is characterized in that the microbial inoculum includes microorganism, binder, dopamine or its salt, the high-polymer molecule.
2. microbial inoculum according to claim 1, which is characterized in that the microbial inoculum further includes following 1) -4) described at least It is a kind of:
1) binder includes that the nutriment of its metabolism or breeding can be provided for the microorganism；Specifically, the binder packet Include starch and/or water；
2) microorganism includes the microorganism beneficial to plant growth；Specifically, the microorganism includes bacillus subtilis, ferment Female bacterium and/or lactic acid bacteria；Again specifically, when the microorganism includes bacillus subtilis, saccharomycete and lactic acid bacteria, withered grass bud Spore bacillus, saccharomycete, lactic acid bacteria mass ratio be 1: 1: 1；
3) the high-polymer molecule includes temperature sensitive the high-polymer molecule；Specifically, the high-polymer molecule includes PNIPAm polymer；
4) dopamine or its salt are covered on the surface of binder and microorganism, and the high-polymer molecule is covered on the DOPA The surface of amine or its salt.
3. according to claim 1 and/or microbial inoculum described in 2, which is characterized in that the microbial inoculum is prepared by the following method:
After microorganism and binder mixing granulation, in the Tris-HCl buffer of pH value 8~8.5, with Dopamine hydrochloride one It rises and 2h or more is stirred at room temperature, after suction filtration, place the product in reaction vessel, add ultrapure water, lead to N2, high polymer is added into solution Monomer, crosslinking agent, the catalyst of molecule are protected from light and continue logical N2, stir at room temperature, then reaction vessel be transferred in ice bath and is stirred It mixes, initiator is added, is transferred in closed mold after stirring, polymerize at room temperature for 24 hours, product is crushed to obtain the final product；
Specifically, by mass, in the method, described microorganism 1-10 parts, 1-10 parts of binder, Dopamine hydrochloride 1-10 Part, 0.8200 part of 10500 parts of monomer, the crosslinking agent of the high-polymer molecule；
It is 7.5 parts of the microorganism, 2.5 parts of binder, 1 part of Dopamine hydrochloride, high in the method again specifically, by mass 1-20 parts of 12-60 parts of monomer, the crosslinking agent of polymers molecule；
Specifically, the binder includes starch and/or water；Specifically, the microorganism includes bacillus subtilis, saccharomycete And/or lactic acid bacteria；Again specifically, when the binder includes starch and water, when the microorganism include bacillus subtilis, When saccharomycete and lactic acid bacteria, bacillus subtilis, saccharomycete, lactic acid bacteria, starch and water mass ratio be 2: 2: 2: 2: 1.
4. a kind of preparation method of microbial inoculum, which is characterized in that the described method includes: by after microorganism and binder mixing granulation, In the Tris-HCl buffer of pH value 8~8.5, it is stirred at room temperature 2h or more together with Dopamine hydrochloride, after suction filtration, product is set In reaction vessel, add ultrapure water, leads to N2, into solution be added the high-polymer molecule monomer, crosslinking agent, catalyst, be protected from light after Continuous logical N2, stir at room temperature, then reaction vessel be transferred in ice bath and is stirred, initiator is added, is transferred to after stirring closed In mold, polymerize at room temperature for 24 hours, product is crushed to obtain the final product.
5. according to the method described in claim 4, it is characterized in that, the method also includes following 1) -6) described at least It is a kind of:
1) binder includes that the nutriment of its metabolism or breeding can be provided for the microorganism；Specifically, the binder packet Include starch and/or water；
2) microorganism includes the microorganism beneficial to plant growth；Specifically, the microorganism includes bacillus subtilis, ferment Female bacterium and/or lactic acid bacteria；Again specifically, when the microorganism includes bacillus subtilis, saccharomycete and lactic acid bacteria, withered grass bud Spore bacillus, saccharomycete, lactic acid bacteria mass ratio be 1: 1: 1；
3) the high-polymer molecule includes temperature sensitive the high-polymer molecule；Specifically, the high-polymer molecule includes PNIPAm polymer；
4) by mass, in the method, described microorganism 1-10 parts, 1-10 parts of binder, 1-10 parts of Dopamine hydrochloride, Gao Ju 0.8-200 parts of 10-500 parts of monomer, the crosslinking agent of object molecule；
Specifically, by mass, in the method, 7.5 parts of the microorganism, 2.5 parts of binder, 1 part of Dopamine hydrochloride, Gao Ju 1-20 parts of 12-60 parts of monomer, the crosslinking agent of object molecule；
Again specifically, the binder includes starch and/or water；Specifically, the microorganism includes bacillus subtilis, yeast Bacterium and/or lactic acid bacteria；Again specifically, when the binder includes starch and water, when the microorganism includes bacillus subtilis When bacterium, saccharomycete and lactic acid bacteria, bacillus subtilis, saccharomycete, lactic acid bacteria, starch and water mass ratio be 2: 2: 2: 2: 1；
5) granulation includes that microorganism and binder are added to High Speed Stirring Machine granulation, and 30-37 DEG C of fluidized drying is to containing Water 3-5% crosses 20 meshes to obtain the final product；
6) by after microorganism and binder mixing granulation, in the Tris-HCl buffer of pH value 8~8.5, with Dopamine hydrochloride It is stirred at room temperature 2h or more together, after suction filtration, place the product in reaction vessel, adds ultrapure water, lead to N2, monomer is added into solution NIPAM, crosslinking agent and catalyst, dosage are respectively that 50ml ultrapure water, NIPAM 11.3g/100g is added in product described in every 1-5g H2O, crosslinking agent clay 0.8-4g/100gH28 μ L/100gH of O, catalyst TEMED2O；It is protected from light and continues logical N2, stir at room temperature Then reaction vessel is transferred in ice bath and stirs 5min by 15min, initiator KPS is added, and KPS dosage is 2g/100g H2O, It is transferred in closed mold, polymerize at room temperature for 24 hours after stirring 30s.
6. the microbial inoculum being prepared by claim 4 and/or 5 the methods.
7. the application of microbial inoculum described in any microbial inoculum of claim 1-3 and/or claim 6.
8. application according to claim 7, which is characterized in that the application includes following 1) -4) described at least one Kind:
1) sustained release or controlled release product as the bacterium of useful plants growth；
2) product as useful plants growth；
3) as Ecological Distribution of Soil Microorganisms or the adjusting product of nutrition condition；
4) as the sustained release of fertilizer or controlled release product.
9. the application of claim 4 and/or 5 the methods.
10. application according to claim 9, which is characterized in that the application includes following 1) -4) described at least one Kind:
1) application in the product or its Related product that preparation has the bacterium of sustained release or the growth of controlled release useful plants；
2) application in the product or its Related product that preparation has useful plants growth；
3) there is the application in the product or its Related product for adjusting Ecological Distribution of Soil Microorganisms or nutrition condition in preparation；
4) application in the product or its Related product that preparation has sustained release or release and release control fertilizer.
CN201810899582.XA 2018-08-08 2018-08-08 Controlled-release agricultural microbial inoculum Active CN109082391B (en)

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