CN110343017A - Polyglutamic acid gel microspherical microbial inoculum and application thereof - Google Patents
Polyglutamic acid gel microspherical microbial inoculum and application thereof Download PDFInfo
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- CN110343017A CN110343017A CN201910788682.XA CN201910788682A CN110343017A CN 110343017 A CN110343017 A CN 110343017A CN 201910788682 A CN201910788682 A CN 201910788682A CN 110343017 A CN110343017 A CN 110343017A
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- polyglutamic acid
- microbial inoculum
- acid gel
- ball
- gel micro
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- 230000003449 preventive effect Effects 0.000 description 1
- 239000006041 probiotic Substances 0.000 description 1
- 235000018291 probiotics Nutrition 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 239000011814 protection agent Substances 0.000 description 1
- 238000005057 refrigeration Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 239000010865 sewage Substances 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 239000002688 soil aggregate Substances 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L sulfate group Chemical group S(=O)(=O)([O-])[O-] QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 230000003519 ventilatory effect Effects 0.000 description 1
- 230000003442 weekly effect Effects 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C21/00—Methods of fertilising, sowing or planting
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G31/00—Soilless cultivation, e.g. hydroponics
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
- A23K20/147—Polymeric derivatives, e.g. peptides or proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K40/00—Shaping or working-up of animal feeding-stuffs
- A23K40/30—Shaping or working-up of animal feeding-stuffs by encapsulating; by coating
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/03—Organic compounds
- A23L29/045—Organic compounds containing nitrogen as heteroatom
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/065—Microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
- A23P10/00—Shaping or working of foodstuffs characterised by the products
- A23P10/30—Encapsulation of particles, e.g. foodstuff additives
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
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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
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- 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
- C05G3/80—Soil conditioners
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- C05G5/00—Fertilisers characterised by their form
- C05G5/30—Layered or coated, e.g. dust-preventing coatings
- C05G5/35—Capsules, e.g. core-shell
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- 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
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- C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
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Abstract
The invention discloses a polyglutamic acid gel microspherical microbial inoculum, which takes microorganisms, gamma-polyglutamic acid modified by glycidyl methacrylate, a photoinitiator and water as disperse phases, hexadecane and span 80 as continuous phases and adopts a droplet microfluidic technology to prepare microspheres, wherein the size of the microspheres is between 100 and 500 mu m. The invention also discloses a preparation method and application of the polyglutamic acid gel microspherical microbial inoculum. The polyglutamic acid gel microspheres can slowly release the embedded microbial inoculum and prolong the effective period of the microbial inoculum with higher effective viable count, have high-activity biological function, are beneficial to colonization of the functional microbial inoculum in soil, can effectively prevent and treat various crop diseases, reduce the use amount of chemical pesticides and chemical fertilizers, effectively combine biological prevention and chemical prevention, make up for deficiencies, realize crop prevention and treatment, and save time and labor.
Description
Technical field
The present invention relates to microorganisms technical fields, and in particular to a kind of polyglutamic acid gel micro-ball microbial inoculum and its application.
Background technique
Soil is the foundation of agricultural planting, and soil issues are concerning national economy.Since reform and opening-up, with China's industry
The rapid development of modernization and Derived from Agricultural Modernization, the especially excessive use, sewage irrigation, continuous cropping management etc. of agriculture chemical
Cause the deterioration of agricultural land soil to be on the rise, and forms the gesture of accumulation year by year.Agricultural land soil deterioration is mainly reflected in soil plate
Knot, the soil salinization, soil flora imbalance, continuous cropping obstacle, heavy metal concentration increase etc..Agricultural land soil deterioration has become system
The about bottleneck problem of agricultural sustainable development.
Studies have shown that soil beneficial microbe can excite soil vitality by the proliferation and secretion activity substance of itself,
Increase soil aggregate structure, make loosing soil, alleviates excessive application chemical fertilizer to the pollution of soil and hardened, can also strongly divide
The microelements such as phosphorus, potassium, iron, silicon that solution, release deposit for many years, increase soil nutrient.Microbial bacterial agent is applied, can be improved soil
Earth beneficial microorganism quantity not only can increase the soil organism and available nutrient, lightened plate floor combined symptoms shape, moreover it is possible to improve root of the crop
Border soil environment inhibits harmful microbe growth, promotes absorption of the crop to nutrient, resists abiotic stress, enhances anti-heavy
Stubble ability, improves crop yield, promotes quality of agricultural product.Therefore, developing microbial manure becomes realization farmland soil modifying
Important promotion means.
China gradually welcome since the beginning of this century microbial manure exploitation upsurge, microbial manure research and development obtain it is considerable into
Exhibition.Although microbial manure market gets prosperous with every passing day, still there are some technical problems urgently to be resolved: 1. microbial activity protective agent
Research lacks, and microbial activity retention time and shelf life are short；2. the biocompatibility and dimensional effect of encapsulated activity protecting agent
Two key factors restrict the development of microbial manure novel form.And the selection and dosage form preparation of microbial activity protection agent material
Technique is the breach that the above problem solves.
Hydrogel material is because having the advantages such as high-moisture, ingredient are similar to extracellular matrix, biocompatibility is good, closely
It is used widely over year in cell and pharmaceutical carrier field.Cell is coated in hydrogel material, only cell does not provide
One three-dimensional bionical microenvironment, the damage that cell can also be avoided suffered in storage and transportational process.But because of traditional water
Gel component is mostly chemical macromolecule, and biocompatibility and degradability are poor, and the use of cross-linking process toxic cross-linking agents is big
The bioactivity of internal package cell is affected greatly；Polyglutamic acid (γ-PGA) is that microorganism utilizes Pidolidone and D-Glu
A kind of anionic polypeptides type polymer that monomer is polymerized by γ-amido bond is one kind of part microorganism itself secretion
Protect pod membrane.In recent years some, can be to avoid food, living thin the study found that γ-PGA is a kind of excellent cell-protecting
There is denaturalization phenomenon in freezing and course of defrosting in born of the same parents and bioactive substance.Bhat et al. is the study found that γ-PGA can be mentioned
Survival rate of the high three kinds of probiotics (L.paracasei, B.longum, B.breve) in freezing dry process.Meanwhile γ-
PGA is also a kind of efficient plant biological stimulin, can significantly improve crop utilization rate of fertilizer, enhances crop anti-adversity ability, mentions
High crop yield and quality.Molasses, and it is commonly called as tangerine water, there are the spies such as nutrition content is high, from a wealth of sources, biological safety is high
Property, this seminar early-stage study, which is shown, can efficiently maintain microbial bacterial agent active.Therefore, γ-PGA and molasses are microorganism guarantors
Protect the ideal chose of agent material.In addition, conventional hydrogels cell carrier size is big, it is unfavorable for the nutrition of all cells in hydrogel
The conveying of substance and metabolite, the cell inside hydrogel can be dead due to the factors such as anoxic, subalimentation.
In the cultivation of crop, various soil-borne diseases for example dead seedling, rotten, it is green it is withered, found withered, withered, root-rot, stem foot
It is rotten, damping off etc. and usually to perplex peasant, solution is mostly applying pesticides or microbial bacterial agent, and because state of the art limits
The defects of system, it is low all to there is effective bacterium number content in the microbial bacterial agent sold currently on the market, effective bacterium easy in inactivation, and after applying
It takes effect slow.It there is no the report with polyglutamic acid sustained-release micro-spheres imbedded microbe microbial inoculum at present, microbial bacterial agent microballoon is anti-
Control also having not been reported for above-mentioned agricultural disease.
Summary of the invention
Technical problem to be solved by the invention is to provide a kind of polyglutamic acid gel micro-ball microbial inoculums, to improve effective bacterium
The time-to-live of content and effective bacterium.
The present invention also technical problems to be solved are to provide the preparation method of above-mentioned polyglutamic acid gel micro-ball microbial inoculum.
The last technical problems to be solved of the present invention are to provide the application of above-mentioned polyglutamic acid gel micro-ball microbial inoculum.
In order to solve the above technical problems, The technical solution adopted by the invention is as follows:
A kind of polyglutamic acid gel micro-ball microbial inoculum, the γ-polyglutamic modified with microorganism, glycidyl methacrylate
Sour (PGA-GMA), photoinitiator and water are dispersed phase, using hexadecane and sorbester p17 as continuous phase, using the micro-fluidic skill of drop
Microballoon is made in art, and the Microsphere Size is between 100-500 μm.
Wherein, the microorganism is preferably A Si Penicillium notatum (P.asturianum), bacillus subtilis (Bacillus
Subtilis), bacillusmusilaginosiengineering (Bacillus mucilaginosus), bacillus amyloliquefaciens (Bacillus
) and any one or the combination of several of them in the general bacterium of camel thorn (Pantoeaalhagi) amyloliquefaciens.Described
Microorganism is more preferably A Si Penicillium notatum (P.asturianum) CGMCC No.17198 and (is recorded in Chinese patent
201910324342.1), bacillus subtilis (Bacillus subtilis) CCTCC NO:M 2016264 (is recorded in China
Patent 201610665832.4), bacillusmusilaginosiengineering (Bacillus mucilaginosus) CCTCC NO:M2016265
(being recorded in Chinese patent 201610665832.4), bacillus amyloliquefaciens (Bacillus amyloliquefaciens)
CCTCC NO:M 2016266 (being recorded in Chinese patent 201610665832.4), the general bacterium of camel thorn (Pantoeaalhagi)
Any one or the combination of several of them in CGMCC NO.15525 (being recorded in Chinese patent 201810815380.2).
It is used further to prepare microbial inoculum after mentioned microorganism fermented and cultured to stationary phase.Bacillus subtilis, gel-shaped gemma bar
Bacterium, bacillus amyloliquefaciens make thallus enter gemma form obtained by high density fermentation by refrigeration, and gemma state is bud
The dormant state of spore bacillus can survive several years in the state of not sprouting and to decades.A Si Penicillium notatum passes through solid state fermentation
Afterwards, brine, 4 layers of lens wiping paper filtering, obtains spore suspension.The general bacterium of camel thorn is after liquid fermentation through being separated by solid-liquid separation
Obtain thallus.
Wherein, the general bacterium fermentation culture conditions of camel thorn are as follows: the general bacterium of camel thorn, which is inoculated in LB culture medium, activates 12-24h,
Activation temperature is 25-35 DEG C, the general bacterium of the camel thorn of activation is inoculated in liquid fermentation medium, 30-35 DEG C of culture 36-50h
Obtain fermentation liquid；It is separated by solid-liquid separation to obtain thallus after fermentation.The fermentative medium formula is as follows: carbon source 10-100g/L, nitrogen source
1-30g/L, inorganic salts 0.01-50g/L, pH5.0-9.0.
Wherein, bacillus subtilis, bacillusmusilaginosiengineering or bacillus amyloliquefaciens fermentation culture conditions are as follows: will be oblique
The Promoting bacteria (bacillus subtilis, bacillusmusilaginosiengineering or bacillus amyloliquefaciens) that face saves is inoculated in activation medium
On, cultivate 18-24h；Strain on picking activation medium, is inoculated in fermentation medium, 30-35 DEG C of culture 12-24h, will
Fermentation liquid or fermentation concentrated liquor are placed in low-temperature treatment 35-55h in Cool Room 4 DEG C；It is separated by solid-liquid separation to obtain spore later.The activation training
Supporting base is LB culture medium, is formulated as follows: peptone 10g/L, yeast powder 5g/L, sodium chloride 10g/L, pH6.0-8.0, the solvent
For water.The formula of the fermentation medium is as follows: carbon source 10-100g/L, nitrogen source 1-40g/L, inorganic salts 0.01-50g/L, solvent
For water, pH5.0-9.0.
Wherein, the fermentation culture conditions of A Si Penicillium notatum are as follows: A Si Penicillium notatum is after solid state fermentation, physiology salt washing
It washs, 4 layers of lens wiping paper filtering obtain spore suspension.Solid-state fermentation culture medium is mixotroph and water, and the mixotrophism is
Wheat bran, tapioca starch, manioc waste, any one or a few in stalk, preferably wheat bran 75g, stalk 25g, water 80mL, culture medium
Sterilize the solid state fermentation in 20cm*40cm venting bags, and inoculum concentration is A Si mould spore suspension 1 × 10720mL, fermentation condition
It is 7 days, 28 DEG C.
The carbon source is glucose, sucrose, maltose, lactose, xylose, fructose, lactic acid, citric acid, glycerol, starch and sugar
Any one or the combination of several of them in honey；The nitrogen source be yeast powder, beef extract, peptone, yeast extract, corn pulp, beancake powder,
Cottonseed meal, urea, (NH4)2SO4、NH4Cl、(NH4)2HPO4And NH4NO3Middle any one or the combination of several of them；It is described inorganic
Salt is sulfate, phosphate, dihydric phosphate, any one or the combination of several of them in phosphoric acid hydrogen disalt and hydrochloride.
Wherein, the polyglutamic acid, molecular weight are 50kDa-3000kDa (preferably 1000kDa)；The polyglutamic
Acid can replace (such as polyglutamic acid sodium salt, polyglutamic acid sylvite) with polyglutamate；The Glycidyl methacrylate is sweet
The gamma-polyglutamic acid of grease modification is prepared as follows to obtain: gamma-polyglutamic acid being dissolved in the water, preferably deionization
In water, adjusting pH value is 4-6 (preferably 4.5), and glycidyl methacrylate is then added dropwise, is stirred at room temperature and is uniformly dissolved, and is dripped
The pH value of control system maintains 4-6 (preferably 4.5) during adding, and is added dropwise under 50-70 DEG C of (preferably 60 DEG C) water-bath
It is stirred to react 6-10 hours (preferably 8 hours).To after reaction, dialysed 3-8 days (preferably 5 days) using bag filter room temperature, most
After be freeze-dried to obtain the final product.Wherein, gamma-polyglutamic acid being dissolved in the water, the quality of water is 10~20 times of polyglutamic acid quality,
The carboxylate radical of gamma-polyglutamic acid and the molar ratio of glycidyl methacrylate are 1:1~1:5.
Wherein, the photoinitiator is phenyl (2,4,6- trimethylbenzoyl) phosphoric acid lithium salts.
In dispersed phase, the ratio that microorganism living bacteria count accounts for water is 107-1010Cfu/mL, methyl propenoic acid glycidyl
The ratio that the gamma-polyglutamic acid of ester modification accounts for water is 1-5g/100mL, and the ratio that photoinitiator accounts for water is 0.1-0.5g/100mL；
In continuous phase, volume ratio shared by sorbester p17 is 1%-10%, remaining is hexadecane.
When the microorganism is A Si Penicillium notatum (P.asturianum), living bacteria count is calculated with spore count, preferably
The ratio of Zhan Shui is 1 × 108-1×1010cfu/mL；When the microorganism is bacillus subtilis (Bacillus
Subtilis), bacillusmusilaginosiengineering (Bacillus mucilaginosus) or bacillus amyloliquefaciens (Bacillus
When amyloliquefaciens), living bacteria count is calculated with gemma, and the ratio for preferably accounting for water is 1 × 108-1×1010cfu/
mL.When the microorganism is the general bacterium of camel thorn (Pantoeaalhagi), living bacteria count is calculated with thallus, preferably accounts for water
Ratio is 1 × 108-1×1010cfu/mL。
Microbial suspension Yu Shuizhong is added methacrylic acid by the preparation method of above-mentioned polyglutamic acid gel micro-ball microbial inoculum
The gamma-polyglutamic acid PBS buffer solution and photoinitiator of ethylene oxidic ester modification, are uniformly mixed and are used as dispersed phase；By hexadecane and
Sorbester p17, which mixes, is used as continuous phase, and the drop of monodispersed submicron scale is made by drop microflow control technique, then solid by light
Change up to polyglutamic acid gel micro-ball microbial inoculum, the Microsphere Size is between 100-500 μm.
Wherein, the polyglutamic acid gel micro-ball microbial inoculum frozen dried that will be obtained.
Wherein, the drop microflow control technique, the design parameter of micro-fluidic chip are as follows: syringe inner diameter is 500-600 μ
M (preferably 550 μm), outer diameter are 900-1000 μm (preferably 960 μm), and injection-tube tip internal diameter is 30-50 μm (preferably 40 μm), outside
Diameter is 40-60 μm (preferably 50 μm)；Receive bore be 100-200 μm (preferably 150 μm), outer diameter be 900-1000 μm (preferably
960μm)；Square tube is 1mm；At 4-8 μ L/min (preferably 6 μ L/min), continuous phase flow control exists dispersed phase flow control
20-40 μ L/min (preferably 30 μ L/min).
Wherein, photo curing condition are as follows: the liquid collected by drop microflow control technique is placed under visible light source, solidifies 1-
3min。
In the polyglutamic acid gel micro-ball microbial inoculum finally prepared, living bacteria count >=6 × 108CFU/mL (preferably >=6 ×
109CFU/mL)。
Microflow control technique technology is accurately to manipulate and handle minute yardstick fluid in microchannel (tens of to hundreds of microns)
Technology, can not only be accurately controlled the size and structure of single emulsion droplet, while have high specific surface area, efficiently
Mass-and heat-transfer characteristic effectively prevents the heat generated in microcapsules forming process to the active influence of microbial inoculum.Based on this, this is specially
Benefit utilizes microflow control technique, intends promoting the degeneration-resistant mechanism of plant growth from natural environment biocontrol bacterial strain and secretion collaboration, with
Wall material based on γ-PGA, microbial bacterial agent are core material, imitative using drop microflow control technique building gamma-polyglutamic acid/microorganism
Raw core-shell microcapsule, by regulate and control microfluidic channel parameter prepare size uniformity, favorable dispersibility, stable structure micro- glue
Capsule bacterial manure, microbial activity in long-effective protection microcapsules solve microbial manure shelf life in production, preservation and application process
Short problem.
Application of the above-mentioned polyglutamic acid gel micro-ball microbial inoculum in fertilizer, feed or field of food is also in protection of the invention
Within the scope of.Preferably, polyglutamic acid gel micro-ball microbial inoculum is as fertilizer in plant disease-resistant growth-promoting, soil phosphorus decomposing, raising agriculture
Crop yield, the application increase soil fertility, enhance any one or a few in crops resistance.
The polyglutamic acid gel micro-ball microbial inoculum can be used cooperatively with conventional chemical fertilizers and pesticide, also can be used as bacterial manure
Directly apply；Polyglutamic acid gel micro-ball microbial inoculum is diluted with water 10-1000 times, by water planting, is sprayed or root irrigation farming
Object.Polyglutamic acid gel micro-ball microbial inoculum of the invention, has the function of high-activity biological, conducive to function stem in the soil quick
It colonizes and is sustained, various crop disease can be effectively prevented, reduce the usage amount of chemical pesticide and chemical fertilizer, be effectively combined life
Object prevention and treatment and chemical prevention, learn from other's strong points to offset one's weaknesses, and realize crop prevention and treatment one, time saving and laborsaving.
The crops be cereal crops (such as rice, wheat, corn etc.), greengrocery crop (such as green vegetables, tomato,
Capsicum etc.) or melon or fruit type crops (such as strawberry, grape, muskmelon, cucumber etc.).
The utility model has the advantages that compared with prior art, the present invention having the advantage that
(1) polyglutamic acid sustained-release micro-spheres provide pod membrane protection in polyglutamic acid gel micro-ball microbial inoculum for disease-resistant Promoting bacteria,
And bacillus is made to form biofilm system, reach a kind of stable equilibrium state, is at gemma state；Polyglutamic acid is by micro-
Biofermentation generates, and is formed by D-Glu and Pidolidone monomer by glutamy amine key homopolymerization, has nontoxic, biological drop
Solution and advantages of environment protection have the absorption and other effects for improving plant to nutrient.
(2) when polyglutamic acid gel micro-ball microbial inoculum is diluted application, polyglutamic acid can provide again for the growth of Promoting bacteria
Nutrition promotes the fast breeding of Promoting bacteria；The gel micro-ball of polyglutamic acid is conducive to keep the vigor of beneficial bacterium, substantially prolongs
The shelf life of polyglutamic acid gel micro-ball microbial inoculum makes the number of viable of the beneficial bacterium in polyglutamic acid gel micro-ball microbial inoculum at 24
>=6 × 10 are maintained in month9CFU/mL。
(3) five kinds of heretofore described microorganisms, crop root microbiologic population knot can be improved by being applied in crops
Structure increases quantity of useful microbe in soil, can improve soil texture, increase soil fertility, keep soil from packing together, Neng Gouti
High crop yield, the anti-adversity abilities such as enhancing crop salt resistance, disease-resistant, cold-resistant.
Detailed description of the invention
Fig. 1 polyglutamic acid gel micro-ball microbial inoculum prepares schematic diagram.
The polyglutamic acid gel micro-ball of microorganism is free of under Fig. 2 microscope.
The gel micro-ball of polyglutamic acid containing P.asturianum under Fig. 3 microscope.
Shadow of Fig. 4 different molecular weight polyglutamic acid to the gel micro-ball microbial inoculum slow-release time of polyglutamic acid containing P.asturianum
It rings.
Influence of tetra- kinds of Fig. 5 different embedding techniques to P.asturianum suspension spore liquid living bacteria count.
Tetra- kinds of Fig. 6 are changed with time using polyglutamic acid gel micro-ball embedding microbial inoculum living bacteria count.
Specific embodiment
According to following embodiments, the present invention may be better understood.However, as it will be easily appreciated by one skilled in the art that real
It applies content described in example and is merely to illustrate the present invention, without sheet described in detail in claims should will not be limited
Invention.
The design parameter for the micro-fluidic chip that following embodiment uses are as follows: syringe inner diameter is 550 μm, and outer diameter is 960 μm,
Injection-tube tip internal diameter is 40 μm, and outer diameter is 50 μm；Receiving bore is 150 μm, and outer diameter is 960 μm；Square tube is 1mm.
The preparation of 1 polyglutamic acid sustained-release micro-spheres of embodiment (being free of microorganism)
It takes 1g (7.8mmol carboxylate radical) gamma-polyglutamic acid (molecular weight 2000kDa) to be dissolved in 20mL deionized water, stirs
It after completely dissolution, is 4.5 with the pH value that dilute hydrochloric acid (0.1wt% concentration) adjusts gamma-polyglutamic acid solution.Then, it is slowly added dropwise
1.02ml (7.8mmol) glycidyl methacrylate, which is stirred at room temperature, to be uniformly dissolved, and uses dilute hydrochloric acid (0.1% during being added dropwise
Concentration) control reaction solution pH value maintain 4.5, be stirred to react under 60 DEG C of water-baths 8 hours.To after reaction, benefit
It is dialysed 5 days with the bag filter room temperature of 3500Da molecule interception, finally freeze-drying obtains the methacrylic acid contracting of white flock
The gamma-polyglutamic acid solid of water glyceride modification.
Sorbester p17 is added in hexadecane to be sufficiently stirred as continuous phase, wherein sorbester p17 accounts for continuous phase total volume
5%.It takes the gamma-polyglutamic acid solid of glycidyl methacrylate modification obtained to be dissolved in deionized water, and light is added and draws
Hair agent phenyl (2,4,6- trimethylbenzoyl) phosphoric acid lithium salts is sufficiently stirred as dispersed phase, glycidyl methacrylate
The accounting of the gamma-polyglutamic acid of modification is 2g/100mL, and the accounting of photoinitiator is 0.5g/100mL.
Two-phase is sucked in the identical syringe of model respectively, be separately fixed on the identical peristaltic pump of bench-types No. two, infused
It is connected between emitter and the pipeline of chip with PE pipe.Dispersed phase mechanical pump is opened, solution is slowly advanced micro-channel device, is adjusted
Continuous phase flow velocity is 20 μ L/min, and dispersed phase flow velocity is 4 μ L/min, it is ensured that stable microballoon is formed in pipeline.Drop is collected
Into centrifuge tube, with radiation of visible light 2-3min, until (Fig. 1-2) is fully cured in polyglutamic acid gel micro-ball.Microsphere Size is big
Generally at 400 μm.
The fermentation of 2 A Si Penicillium notatum of embodiment
Microbial bacterial agent of the present invention is prepared as follows:
Wherein, the fermentation culture conditions of A Si Penicillium notatum are as follows: A Si Penicillium notatum CGMCC No.17198 is sent out by solid-state
After ferment, brine, 4 layers of lens wiping paper filtering obtain spore suspension.Solid-state fermentation culture medium be mixotroph and
Water, the mixotrophism are wheat bran, tapioca starch, manioc waste, any one or a few in stalk, preferably wheat bran 75g, stalk
25g, water 80mL, medium sterilization solid state fermentation in 20cm*40cm venting bags, inoculum concentration are A Si mould spore suspension 1
×10720mL, fermentation condition are 7 days, 28 DEG C.Spore suspension is made with spore under sterile washing, through blood counting chamber meter
Number, spore suspension miospore number are 1 × 104~1 × 108CFU/mL。
The fermentation of 3 camel thorn of embodiment general bacterium, bacillus subtilis, bacillusmusilaginosiengineering, bacillus amyloliquefaciens
The general bacterium CGMCC NO.15525 of camel thorn that 4 DEG C of inclined-planes save is inoculated in eggplant shape culture bottle culture medium (to activate
Culture medium) on, 30 DEG C of culture 16h.Strain on picking activation medium, is inoculated in fermentation medium, 30 DEG C, cultivates 40h,
Fermentation liquid or fermentation concentrated liquor are placed in low-temperature treatment 48h in Cool Room 4 DEG C, is coated with and measures through plate, the general bacterium hair of camel thorn
Number of viable in zymotic fluid can reach 1 × 1010CFU/mL or more.Culture medium in the eggplant shape culture bottle is LB culture medium；Hair
Ferment nutrient media components are as follows: sucrose 50g/L, beef extract 3g/L, NaCl 8g/L, peptone 8g/L, pH 6.0-9.0.
Bacillus subtilis CCTCC NO:M 2016264, the bacillusmusilaginosiengineering CCTCCNO:M that 4 DEG C of inclined-planes are saved
2016265, bacillus amyloliquefaciens CCTCC NO:M 2016266 is activated, and it is (i.e. living to be inoculated in eggplant shape culture bottle culture medium respectively
Change culture medium) on, 30 DEG C of culture 18h.The strain grown is transferred in fermentation medium respectively, after 30 DEG C of cultures for 24 hours, will be sent out
Zymotic fluid is placed in low-temperature treatment 48h in Cool Room 4 DEG C, is coated with and measures through plate, and the number of viable in every kind of microbial fermentation solution is reachable
To 1 × 1010CFU/mL or more.Culture medium in the eggplant shape culture bottle is LB culture medium, fermentation medium component are as follows: glucose
50g/L, peptone 10g/L, (NH4)2SO40.05g/L, NaCl1g/L, MnSO4·4H2O 0.5g/L, KH2PO41.0g/L
PH is 6.0-9.0.
The ventilatory capacity of filtrated air is 0.6vvm during the fermented and cultured of above-described production tank, and mixing speed is
220rpm。
The preparation of the 4 gel micro-ball microbial inoculum of polyglutamic acid containing P.asturianum of embodiment
It is added what 2% embodiment 2 obtained in polyglutamic acid (molecular weight 1000kDa) solution containing photoinitiator
P.asturianum microbial inoculum, the ratio that microorganism living bacteria count accounts for water is 108Cfu/mL, glycidyl methacrylate are repaired
The ratio that the gamma-polyglutamic acid of decorations accounts for water is 3g/100mL, and the ratio that photoinitiator accounts for water is 0.3g/100mL；In continuous phase,
Volume ratio shared by sorbester p17 is 5%, remaining is hexadecane.Repetition prepares the operation of polyglutamic acid gel micro-ball, is made and contains
P.asturianum polyglutamic acid gel micro-ball microbial inoculum, wherein viable count is about 107cfu/mL.(Fig. 3).
5 different molecular weight polyglutamic acid of embodiment is to the gel micro-ball microbial inoculum slow-release time of polyglutamic acid containing P.asturianum
Influence
Polyglutamic acid molecular size range is a key factor for influencing microsphere sustained-release.Microbial inoculum preparation is embedded with embodiment 4
For technique, the difference is that select molecular weight be respectively 50kDa, 100kDa, 500kDa, 1000kDa, 2000kDa,
The polyglutamic acid of 3000kDa is made the P.asturianum microballoon that partial size is 200 μm, is delayed at 25 DEG C as microsphere supported
Release experiment and using plate count method.As a result see Fig. 4.
It is drawn a conclusion by experiment, polyglutamic acid molecular weight is smaller, and P.asturianum microballoon discharges living bacteria count more
Fastly；Molecular weight is bigger, and it is slower that microballoon discharges living bacteria count.The microsphere sustained-release time made of the polyglutamic acid of 50kDa molecular weight
Most short, complex micro organism fungicide sustained-release micro-spheres sustained release rate made of 1000kDa polyglutamic acid sustained-release micro-spheres can satisfy in not
It with the demand of crop and different stages of growth, applies the microbial inoculum and can discharge for first day and reach 30%, it is sufficient in crop soil
The middle dominant strain for establishing the bacterial strain, and can be sustained and reach 15 days.
6 different-grain diameter P.asturianum polyglutamic acid gel micro-ball microbial inoculum stability of embodiment compares
It is added what 2% embodiment 2 obtained in polyglutamic acid (molecular weight 1000kDa) solution containing photoinitiator
P.asturianum microbial inoculum, the ratio that microorganism living bacteria count accounts for water is 108cfu/mL, and glycidyl methacrylate is repaired
The ratio that the gamma-polyglutamic acid of decorations accounts for water is 3g/100mL, and the ratio that photoinitiator accounts for water is 0.5g/100mL；In continuous phase,
Volume ratio shared by sorbester p17 is 5%, remaining is hexadecane.
Two-phase is sucked in the identical syringe of model respectively, repeat the operation of polyglutamic acid gel micro-ball, preparation is different
The microballoon microbial inoculum of partial size, related process are as follows:
(1) continuous phase flow velocity is 30 μ L/min, and dispersed phase flow velocity is 3 μ L/min, and Microsphere Size is probably at 100 μm.
(2) continuous phase flow velocity is 30 μ L/min, and dispersed phase flow velocity is 6 μ L/min, and Microsphere Size is probably at 400 μm.
(3) continuous phase flow velocity is 30 μ L/min, and dispersed phase flow velocity is 15 μ L/min, and Microsphere Size is probably at 500 μm.
Two-phase is sucked in the identical syringe of model respectively, be separately turned on dispersed phase mechanical pump, solution is slowly advanced
Mixing duct adjusts continuous phase flow velocity and dispersed phase flow velocity, it is ensured that stable gel ball is formed in pipeline.By drop collect to from
In heart pipe, with radiation of visible light 5min, until polyglutamic acid gel micro-ball is fully cured.Particularly relevant technique is as follows:
(1) continuous phase flow velocity is 1mL/min, and dispersed phase flow velocity is 0.2mL/min, and gel ball size is probably in 300mm.
(2) continuous phase flow velocity is 1mL/min, and dispersed phase flow velocity is 0.1mL/min, and gel ball size is probably in 100mm.
Various sizes of gel ball is stood 3 months with physiological saline respectively, during which samples observation weekly, finds size
The gel ball of 100mm and 300mm 7 days there have been part reunite, by cell anyway dye, discovery size 100mm and
The P.asturianum embedded in the gel ball of 300mm accounts for 30% or more in the 28th day dead cell, and 100 μm, 400 μm and 500 μ
Gel micro-ball 3 months of m size remain stable ball-type capsule structure, and living cells is not less than 85%, utilizes micro-fluidic skill
100 μm of -500 μm of gel micro-ball stability of art preparation significantly increase.
The 7 gel micro-ball microbial inoculum of polyglutamic acid containing P.asturianum of embodiment can open preservation up to 24 months
Microbial inoculum used in the present embodiment is as follows:
(1) A Si Penicillium notatum suspension spore liquid in embodiment 2 is diluted to living bacteria count is 7 × 108Cfu/mL, as
Microbial inoculum 1；
(2) A Si Penicillium notatum suspension spore liquid in embodiment 2 is diluted to living bacteria count is 7 × 108Cfu/mL is used in combination
(Technology origin: [Chinese invention, Chinese invention authorization] CN201510565088.6 one kind is micro- for chitosan and sodium tripolyphosphate embedding
Biological compound fungi agent and its preparation method and application), as microbial inoculum 2；
(3) A Si Penicillium notatum suspension spore liquid in embodiment 2 is diluted to living bacteria count is 7 × 108Cfu/mL, with ten
Compound coagulating bath A embedding that sodium dialkyl sulfate, sodium tripolyphosphate and sodium alginate three are formed (Technical Reference:
A kind of preparation method of multilayer embedding microcapsules of CN201710892119.8), as microbial inoculum 3；
(4) microbial inoculum for preparing embodiment 4 is as microbial inoculum 4.
Above-mentioned 4 kinds of microbial inoculums are placed in 25 DEG C, is stored in the environment of relative humidity 40%, 1mL microbial inoculum was respectively taken every 2 months,
Dilution 108Times, it in applying solid LB culture medium, is placed under 32 DEG C of environment, cultivates 18-24h, obtained using colony counting method effective
Number of viable, continuous sampling 26 months.If three groups of parallel laboratory tests, living bacteria count is changed over time such as Fig. 5 institute in different microbial inoculums
Show.
As shown in Figure 5, microbial inoculum 1, microbial inoculum 2,3 living bacteria count >=6 × 10 of microbial inoculum7CFU/mL can maintain respectively 10 months,
The living bacteria count amount of 16 months, 18 months, microbial inoculum 4 reached >=6 × 10 before 24 months7The level of CFU/mL.Thus may be used
See, validity period of polyglutamic acid gel micro-ball microbial inoculum of this patent protection is 24 months, better than common sodium alginate and
Chitosan imbedded processing.
Influence of the 8 polyglutamic acid gel micro-ball of embodiment to different microbial inoculum validity periods
Microbial inoculum used in the present embodiment is as follows:
(1) 7 × 10 are diluted to the general bacterium of camel thorn in embodiment 3 in the present embodiment8Cfu/mL, referring to the side of embodiment 4
Method is embedded with polyglutamic acid gel micro-ball, as microbial inoculum a；
(2) 7 × 10 are diluted to bacillusmusilaginosiengineering in embodiment 3 in the present embodiment8Cfu/mL, referring to embodiment 4
Method embedded with polyglutamic acid gel micro-ball, as microbial inoculum b；
(3) 7 × 10 are diluted to bacillus amyloliquefaciens in embodiment 3 in the present embodiment8Cfu/mL, referring to embodiment 4
Method embedded with polyglutamic acid gel micro-ball, as microbial inoculum c；
(4) 7 × 10 are diluted to bacillus subtilis in embodiment 3 in the present embodiment8Cfu/mL, referring to embodiment 4
Method is embedded with polyglutamic acid gel micro-ball, as microbial inoculum d；
Above-mentioned 4 kinds of microbial inoculums are placed in 25 DEG C, is stored in the environment of relative humidity 40%, 1mL microbial inoculum was respectively taken every 2 months,
Dilution 108Times, it in applying solid LB culture medium, is placed under 32 DEG C of environment, cultivates 18-24h, obtained using colony counting method effective
Number of viable, continuous sampling 26 months.If three groups of parallel laboratory tests, living bacteria count is changed over time such as Fig. 6 institute in different microbial inoculums
Show.
It will be appreciated from fig. 6 that microbial inoculum a living bacteria count amount >=5 × 107CFU/mL is maintained 18 months, microbial inoculum b, microbial inoculum c, microbial inoculum d
Living bacteria count can reach 24 months standards and reach >=6 × 107The level of CFU/mL.It can be seen that polyglutamic acid
Gel micro-ball can effectively extend the holding time of the higher living bacteria count of microbial bacterial agent.
The 9 gel micro-ball microbial inoculum of polyglutamic acid containing P.asturianum of embodiment can reduce the application of chemical fertilizer
The polyglutamic acid gel micro-ball microbial inoculum containing P.asturianum in the present embodiment selects embodiment 4 to prepare microbial inoculum.
Using potting cucumber as experimental subjects, test sets 3 processing, 3 repetitions of each processing, wherein processing 1 is experimental control common fertilizer
Material, processing 2 are common fertilizer+polyglutamic acid gel micro-ball microbial inoculum, and processing 3 is to subtract the common fertilizer+polyglutamic for applying 40% mass
Acid gel microballoon microbial inoculum.The fertilizer applied is every kilogram of soil 150mg, and the dosage of polyglutamic acid gel micro-ball microbial inoculum is every kilogram
Native 0.2mL, the results are shown in Table 2.
Influence of the 2 polyglutamic acid gel micro-ball microbial inoculum of table to potting cucumber growth characteristic and yield
Polyglutamic acid gel micro-ball microbial inoculum in the present invention containing P.asturianum is mixed with fertilizer to be applied, and cucumber root can be promoted
The development in portion makes root thickening, increases, and has apparent promotion to make cucumber plant plant height, underground part fresh weight, overground part fresh weight
With total fresh weight can increase by 26.9%.When subtracting the fertilizer for applying 40% mass, still 11.5% can be improved to cucumber yield, this
Illustrate polyglutamic acid in the present invention, biological organic matter, growth promoting bacteria agent polyglutamic acid gel micro-ball microbial inoculum plant can be promoted raw
It is long, and effectively reduce the usage amount of chemical fertilizer.
The 10 gel micro-ball microbial inoculum of polyglutamic acid containing P.asturianum of embodiment can reduce the application of pesticide
The polyglutamic acid gel micro-ball microbial inoculum preparation containing P.asturianum in the present embodiment selects embodiment 4 to prepare bacterium
Agent.Using potting Xiaoqinling Nature Reserve as experimental subjects, test sets 3 processing, 3 repetitions of each processing, wherein processing 1 is test nitre sodium
Methyl α-naphthyl acetate, processing 2 are nitre sodium methyl α-naphthyl acetate+polyglutamic acid gel micro-ball microbial inoculum, and processing 3 is to subtract the nitre sodium naphthalene for applying 40% volume
Acetic acid+polyglutamic acid gel micro-ball microbial inoculum.The pesticide applied is that every kilogram of soil sprays 2000 times of liquid 50ml, polyglutamic acid gel
The dosage of microballoon microbial inoculum is every kilogram of soil 0.2mL, the results are shown in Table 3.
Influence of the 3 gel micro-ball microbial inoculum of polyglutamic acid containing P.asturianum of table to potting Xiaoqinling Nature Reserve growth characteristics and yield
Polyglutamic acid gel micro-ball microbial inoculum in the present invention containing P.asturianum is mixed with fertilizer to be applied, and Xiaoqinling Nature Reserve can be promoted
The development of root and blade increases root thickening, blade, all to Xiaoqinling Nature Reserve plant plant height, underground part fresh weight, overground part fresh weight
There is apparent facilitation, 26.9% can be increased production.It, still can be to Xiaoqinling Nature Reserve output increased when subtracting the fertilizer for applying 40% mass
13.2%, this illustrates that polyglutamic acid gel micro-ball microbial inoculum can promote plant growth in the present invention, and effectively reduces chemistry
The usage amount of pesticide.
The 11 significant controlling plant diseases of the gel micro-ball microbial inoculum of polyglutamic acid containing P.asturianum of embodiment
Polyglutamic acid gel micro-ball microbial inoculum in the present embodiment containing P.asturianum selects embodiment 4 to prepare microbial inoculum.With
Pot-strawberry is experimental subjects, and test sets four processing altogether, and each processing is arranged 3 in parallel.Processing T1 be control group, successively plus
Enter equal amount of distilled water, processing T2 is first to add equal amount of distilled water, Fusariun oxysporun 105/ mL spore suspension infects
Strawberry, processing T3 are first to apply equivalent polyglutamic acid gel micro-ball microbial inoculum dilution, and Fusariun is added after three weeks
oxysporun 105/ mL spore suspension infects strawberry, and processing T4 is only to add equivalent polyglutamic acid gel micro-ball microbial inoculum
Dilution, strawberry management are daily management, and calculate disease index and opposite control efficiency.Sick grade is as follows: 0: not falling ill；1: disease
The 0~25% of the total radical of radical Zhan；The 25~50% of 2: old complaint number Zhan total radical；The 50~75% of 3: old complaint number Zhan total radical；
The 75~100% of 4: old complaint number Zhan total radical.Each processing Strawberry Root Rot is investigated according to the grade scale of Strawberry Root Rot
The state of an illness calculates disease index.
Z, X and N respectively indicate disease index, each sick grade strain number and the total root segment number of investigation sample in formula；10 be highest disease grade
Strawberry Root Rot is calculated according to formulaWherein DIT2It is processing 2
Disease index, DIT3It is the disease index for handling 3
Using one-way analysis of variance, and carry out F inspection.It the results are shown in Table 4.
Preventive and therapeutic effect of the 4 gel micro-ball microbial inoculum of polyglutamic acid containing P.asturianum of table to Strawberry Root Rot
Polyglutamic acid gel micro-ball microbial inoculum in the present invention containing P.asturianum is mixed with fertilizer to be applied, and strawberry root is effectively inhibited
The disease incidence of maize ear rot, and can be improved the yield of strawberry.T3 is handled compared with handling T2, polyglutamic acid gel micro-ball microbial inoculum is effective
The illness rate for reducing strawberry is reduced to 4.4% from 45.3%.It compares from processing T4 with T1 as it can be seen that in normal strawberry
In, complex micro organism fungicide sustained-release micro-spheres also play the effect of diseases prevention, growth-promoting, and strawberry fresh weight is made to improve 42.1%.
Test result shows that the polyglutamic acid gel micro-ball microbial inoculum in the present invention not only can effectively prevention and control Strawberry Root Rot be done harm to, and
The yield of strawberry can also be significantly improved.
Claims (10)
1. a kind of polyglutamic acid gel micro-ball microbial inoculum, which is characterized in that modified with microorganism, glycidyl methacrylate
Gamma-polyglutamic acid, photoinitiator and water are dispersed phase, using hexadecane and sorbester p17 as continuous phase, using the micro-fluidic skill of drop
Microballoon is made in art, and the Microsphere Size is between 100-500 μm.
2. polyglutamic acid gel micro-ball microbial inoculum according to claim 1, which is characterized in that the microorganism is A Siqing
Mould (P.asturianum), bacillus subtilis (Bacillus subtilis), bacillusmusilaginosiengineering (Bacillus
Mucilaginosus), bacillus amyloliquefaciens (Bacillus amyloliquefaciens) and the general bacterium of camel thorn
(Pantoeaalhagi) any one or the combination of several of them in.
3. polyglutamic acid gel micro-ball microbial inoculum according to claim 1, which is characterized in that the polyglutamic acid, point
Son amount is 50kDa-3000kDa；The polyglutamic acid is replaced with polyglutamate；The glycidyl methacrylate
The gamma-polyglutamic acid of modification is prepared as follows to obtain: gamma-polyglutamic acid is dissolved in the water, adjusting pH value is 4-6,
Then glycidyl methacrylate is added dropwise, is stirred at room temperature and is uniformly dissolved, the pH value of control system maintains during dropwise addition
4-6 is added dropwise under 50-70 DEG C of water-bath and is stirred to react 6-10 hours.To after reaction, be dialysed using bag filter room temperature
It 3-8 days, is finally freeze-dried to obtain the final product.
4. polyglutamic acid gel micro-ball microbial inoculum according to claim 1, which is characterized in that the photoinitiator is phenyl
(2,4,6- trimethylbenzoyl) phosphoric acid lithium salts.
5. polyglutamic acid gel micro-ball microbial inoculum according to claim 1, which is characterized in that in dispersed phase, microorganism is effective
The ratio that viable count accounts for water is 107-1010The gamma-polyglutamic acid of cfu/mL, glycidyl methacrylate modification account for the ratio of water
Example is 1~5g/100mL, and the ratio that photoinitiator accounts for water is 0.1-0.5g/100mL；In continuous phase, volume ratio shared by sorbester p17
For 1%-10%, remaining is hexadecane.
6. the preparation method of polyglutamic acid gel micro-ball microbial inoculum described in claim 1, which is characterized in that by microbial suspension in
In water, the gamma-polyglutamic acid PBS buffer solution and photoinitiator of addition glycidyl methacrylate modification are uniformly mixed and make
For dispersed phase；Hexadecane and sorbester p17 are mixed and are used as continuous phase, monodispersed submicron ruler is made by drop microflow control technique
The drop of degree, then by photocuring up to polyglutamic acid gel micro-ball microbial inoculum, the Microsphere Size is between 100-500 μm.
7. preparation method according to claim 6, which is characterized in that the drop microflow control technique, micro-fluidic chip
Design parameter are as follows: syringe inner diameter be 500-600 μm, outer diameter be 900-1000 μm, injection-tube tip internal diameter be 30-50 μm,
Outer diameter is 40-60 μm；Receiving bore is 100-200 μm, and outer diameter is 900-1000 μm；Square tube is 1mm；Dispersed phase flow velocity
Control is in 4-8 μ L/min, and continuous phase flow control is in 20-40 μ L/min.
8. preparation method according to claim 6, which is characterized in that photo curing condition are as follows: the micro-fluidic skill of drop will be passed through
The liquid that art is collected is placed under visible light source, solidifies 1-3min.
9. application of the polyglutamic acid gel micro-ball microbial inoculum described in claim 1 in fertilizer, feed or field of food.
10. application according to claim 9, which is characterized in that polyglutamic acid gel micro-ball microbial inoculum is as fertilizer in plant
Disease-resistant growth-promoting, improves crop yield, increases soil fertility, enhances any one in crops resistance or several at soil phosphorus decomposing
The application of kind.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110922254A (en) * | 2019-12-20 | 2020-03-27 | 付洪斌 | Agricultural rare earth containing precipitation inhibitor and preparation method and application thereof |
| CN111248196A (en) * | 2020-03-26 | 2020-06-09 | 南京工业大学 | Cell membrane bionic agricultural microbial agent microcapsule and preparation method thereof |
| CN111777459A (en) * | 2020-07-28 | 2020-10-16 | 南京轩凯生物科技有限公司 | Composite synergist and preparation method and application thereof |
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Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102763684A (en) * | 2011-05-01 | 2012-11-07 | 华中农业大学 | Microcapsule preparation, preparation method and application |
| CN103373870A (en) * | 2012-04-12 | 2013-10-30 | 武汉瑞阳生物科技有限公司 | Composite microbial fertilizer and preparation method therefor |
| CN108147893A (en) * | 2017-12-28 | 2018-06-12 | 山东胜景旅游发展有限公司 | Alkaline land modifying agent prepared using bacteria agent and preparation method thereof |
| CN108191574A (en) * | 2018-02-28 | 2018-06-22 | 河北百禾丰化肥有限公司 | A kind of complex microorganism multielement coating enhanced fertilizer and its manufacturing method |
| CN108794187A (en) * | 2018-07-24 | 2018-11-13 | 南京轩凯生物科技有限公司 | A kind of microbial liquid microbial inoculum and its preparation method and application |
| CN109045354A (en) * | 2018-07-26 | 2018-12-21 | 华中科技大学 | A kind of formed in situ injection aquagel for bone-cartilage comprehensive repair |
| CN109516838A (en) * | 2019-01-30 | 2019-03-26 | 韩亚博 | A kind of water conservation Synergistic composite fertilizer and preparation method thereof |
| CN109957517A (en) * | 2019-04-22 | 2019-07-02 | 南京工业大学 | A kind of A Si Penicillium notatum and its microbial inoculum and application |
-
2019
- 2019-08-26 CN CN201910788682.XA patent/CN110343017A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102763684A (en) * | 2011-05-01 | 2012-11-07 | 华中农业大学 | Microcapsule preparation, preparation method and application |
| CN103373870A (en) * | 2012-04-12 | 2013-10-30 | 武汉瑞阳生物科技有限公司 | Composite microbial fertilizer and preparation method therefor |
| CN108147893A (en) * | 2017-12-28 | 2018-06-12 | 山东胜景旅游发展有限公司 | Alkaline land modifying agent prepared using bacteria agent and preparation method thereof |
| CN108191574A (en) * | 2018-02-28 | 2018-06-22 | 河北百禾丰化肥有限公司 | A kind of complex microorganism multielement coating enhanced fertilizer and its manufacturing method |
| CN108794187A (en) * | 2018-07-24 | 2018-11-13 | 南京轩凯生物科技有限公司 | A kind of microbial liquid microbial inoculum and its preparation method and application |
| CN109045354A (en) * | 2018-07-26 | 2018-12-21 | 华中科技大学 | A kind of formed in situ injection aquagel for bone-cartilage comprehensive repair |
| CN109516838A (en) * | 2019-01-30 | 2019-03-26 | 韩亚博 | A kind of water conservation Synergistic composite fertilizer and preparation method thereof |
| CN109957517A (en) * | 2019-04-22 | 2019-07-02 | 南京工业大学 | A kind of A Si Penicillium notatum and its microbial inoculum and application |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110922254A (en) * | 2019-12-20 | 2020-03-27 | 付洪斌 | Agricultural rare earth containing precipitation inhibitor and preparation method and application thereof |
| CN111248196A (en) * | 2020-03-26 | 2020-06-09 | 南京工业大学 | Cell membrane bionic agricultural microbial agent microcapsule and preparation method thereof |
| CN111777459A (en) * | 2020-07-28 | 2020-10-16 | 南京轩凯生物科技有限公司 | Composite synergist and preparation method and application thereof |
| CN112155124A (en) * | 2020-10-15 | 2021-01-01 | 南京轩凯生物科技股份有限公司 | Feed additive containing mixed nucleotide and preparation method thereof |
| CN113174387A (en) * | 2021-05-13 | 2021-07-27 | 浙江海洋大学 | Hydrogel immobilized microorganism preservation method |
| CN113748908A (en) * | 2021-08-17 | 2021-12-07 | 海南农佳佳科技发展有限公司 | Green planting method for pecans |
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