CN109497050A - A kind of Paenibacillus polymyxa fermentation material and its preparation method and application - Google Patents
A kind of Paenibacillus polymyxa fermentation material and its preparation method and application Download PDFInfo
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- CN109497050A CN109497050A CN201811435372.1A CN201811435372A CN109497050A CN 109497050 A CN109497050 A CN 109497050A CN 201811435372 A CN201811435372 A CN 201811435372A CN 109497050 A CN109497050 A CN 109497050A
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- paenibacillus polymyxa
- sporinite
- fermentation material
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/02—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing liquids as carriers, diluents or solvents
- A01N25/04—Dispersions, emulsions, suspoemulsions, suspension concentrates or gels
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/26—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests in coated particulate form
- A01N25/28—Microcapsules or nanocapsules
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/10—Animals; Substances produced thereby or obtained therefrom
Abstract
The invention discloses a kind of Paenibacillus polymyxa fermentation materials and its preparation method and application.The preparation method of the fermentation material include: strain is successively activated, seed culture and fermented and cultured, after being centrifuged, being concentrated, obtain Paenibacillus polymyxa fermentation material；The fermentation material is the mixture of mycoderm and Paenibacillus polymyxa sporinite；The strain is Paenibacillus polymyxa (Paenibacillus poly myxa) ZJU-N.The present invention is by selecting the component type and dosage of specific Paenibacillus polymyxa, change seed culture medium and fermentation medium to ferment, it can obtain the tunning rich in mycoderm and Paenibacillus polymyxa sporinite, and it is used to prepare sporinite micro-capsule and sporinite microcapsule suspending agent, prevent and treat the banded sclerotial blight, false smut and bakanae disease of rice.
Description
Technical field
The present invention relates to biological control on crop disease technical field more particularly to a kind of Paenibacillus polymyxa fermentation materials
And its preparation method and application.
Background technique
Biological pesticide refers to: " being carried out using biological living or its metabolite to harmful organisms such as pest, germ, weeds
A kind of pesticidal preparations of prevention and treatment, or the biological products by biotechnology or bio-mimetic syntheses with pesticide function ".In pesticide
Biological pesticide concept is usually narrow sense in registration management, refers mainly to Natural Enemies of Insects, botanical pesticide, microbial pesticide and turns base
Because of biology.
A few days ago, though domestic chemical pesticide has played important function, and problem of environmental pollution is more and more prominent, biological pesticide
Green Development is the needs of grain security, ecological safety and Environmental security, while can be further reduced disease pest and weed drug resistance
Occur.
Paenibacillus polymyxa (Paenibacillus polymyxa) is a kind of sporiferous gram-positive bacterium, class
Bacillus, widely distributed, there is only soil, root system of plant and body surfaces, and are also that interior life common in plant is thin
Bacterium.Paenibacillus polymyxa has non-pathogenic to plant, while having diseases prevention and growth-promoting functions, is widely applied as biocontrol microorganisms
In agricultural production.Paenibacillus polymyxa quickly can be colonized and be moved in plant host, so as to compete with pathogen
Space and nutrition, it can also improve the disease resistance of host plant by inducing action, and be place by fixed nitrogen and Soluble phosphorus effect
Main plant provides ingredient, promotes plant growth.
Natural biological film (mycoderm): it is a variety of that Paenibacillus polymyxa can generate peptides, protein-based, ucleosides and phenols etc.
Antibacterial material can prevent and treat a variety of plant diseases as caused by pathogenetic bacteria, disease fungus, root-knot nematode, while can form viscosity
On the one hand mycoderm, being formed for mycoderm can be controlled by the growth and breeding for adjusting microbial cells, on the other hand can also be with
It is controlled by adjusting the secretion of exocellular polysaccharide in microorganism growth process, S layers of albumen.Mycoderm acts on bacterium or sporinite,
It is easy to be attached to various target body surfaces and favorably colonizes, environment resistance can be improved while being conducive to spore germination, in antibacterial work
Property etc. many-sided also play an important role.
Artificial membrane (liposome): can form multilayer micro-capsule when phosphatide is dispersed in water, and each layer is that phosphatide is double
Molecular layer is separated by water phase between each layer, and this phospholipid bilayer structure with similar biomembrane is known as liposome.Due to it
Structure be similar to cell, so also known as artificial membrane.Liposome has lasting effect as the carrier of drug.Liposome
As the storage cavern of drug, it can make to be encapsulated drug slow release, to make pharmaceutical preparation that there is controlled-release function.
The raw material for preparing liposome is mainly lipid material, using more based on vegetal soybean lecithin,
Predominantly unsaturated fatty acid can make liposome have lower phase transition temperature, there is good mobility and low viscous
Property, while soybean lecithin is as amophoteric surface active substance, non-toxic, is influenced by pH value, salinity smaller, and biology is simultaneous
Capacitive is good, and itself also has good physiological activity, and the saturating mucosal absorption of rush property drug can independently form micro emulsion.
Conventional liposome has the advantages that many uniquenesses, however, there are also certain as a kind of good carrier
Deficiency, over time, there is content infiltration equistability caused by flocculation, sedimentation, partial size increase is poor
Problem limits its application in medicine, pesticide industry.Many drugs are spraying application to field, peak value are reached in the short time, i.e.,
It is degraded.Since the pharmaceutical carrier stability of high molecular polymer is good, modified liposome surface is gone to it, passes through surface modification mistake
Liposome microbial medicine is wrapped up, the discharge and metabolism of drug can be delayed, while can effectively extend microorganism medicine
Residence time of the object in plant, to extend drug effect.On the other hand, it is good fixed to be able to maintain microorganism for modified liposome
Grow ability.
Polyethylene glycol (PEG) Coated Liposomes: PEG is the polyethers high polymer being made of ethoxy unit, is a kind of nothing
Malicious, without side-effects, environmental-friendly water soluble polymer.Liposome can be improved as adhesive in PEG-4000
The surface filming and solidification intensity of body, are conducive to the lasting effect of drug release in utricule, are also advantageous that in holding microorganism spore
The microenvironment of daughter sprouting vigor.Meanwhile PEG have it is cheap, be easy to get, be not only dissolved in water be but also dissolved in a variety of organic solvents, had solely
Special end group reactivity and modifiability.
Summary of the invention
The object of the present invention is to provide a kind of Paenibacillus polymyxa fermentation material and its preparation method and application, the systems
The fermentation material that Preparation Method obtains is rich in sporinite and mycoderm, can be used for preparing sporinite micro-capsule and sporinite microcapsule suspending agent, and
For preventing and treating the banded sclerotial blight, false smut and bakanae disease of rice.
Specific technical solution is as follows:
The present invention provides a kind of preparation methods of Paenibacillus polymyxa fermentation material, comprising: successively carries out the work of strain
Change, seed culture and fermented and cultured, after being centrifuged, being concentrated, obtain Paenibacillus polymyxa fermentation material；The fermentation material is mycoderm
With the mixture of Paenibacillus polymyxa sporinite；
The strain is named as Paenibacillus polymyxa (Paenibacillus polymyxa) ZJU-N, deposit number
Are as follows: CCTCC NO:M2018692, the deposit date is on October 19th, 2018；
The medium component of the seed culture are as follows: sucrose 10-15g/L, tryptone 5-10g/L, yeast extract 3-
5g/L, beef extract 1.5-3g/L, NaCl 8-10g/L, pH 7.0-7.2；
The medium component of the fermented and cultured are as follows: potato 220-250g/L, tryptone 5-8g/L, sucrose 10-
15g/L, yeast extract 1-5g/L, beef extract 0.5-1g/L, NaCl 1-5g/L, Ca (NO3)2·4H2O 0.5-2g/L,
Na2HPO4.12H2O 1.5-3g/L, pH7.0-7.2.
Paenibacillus polymyxa (Paenibacillus polymyxa) ZJU-N used in above-mentioned preparation method is rice
Multi-functional endophyte, parasitism in rice, can quickly occupy advantageous ecological niche, and can establish with host plant in rice
Symbiosis, while can effectively inhibit infecting for rice multiple diseases, disease incidence is reduced, the production of rice is significantly improved
Amount and safety, realize the healthy No-harmful apple orchard of rice.Paenibacillus polymyxa (Paenibacillus polymyxa)
ZJU-N also has the speed of growth fast, produces the strong feature of spore ability；Importantly, the bacterial strain can high yield mycoderm, after being conducive to
The preparation of continuous sporinite micro-capsule and suspending agent, improves the control efficiency of suspending agent.
The present invention carries out fermentation process using above-mentioned specific Paenibacillus polymyxa bacterial strain, and by changing seed culture
The component of the raw material type and other each nutrients in the source C and the source N, can make Paenibacillus polymyxa exist in base and fermentation medium
The trophosome of seed liquor and fermented and cultured bacterial strain early period exponentially type rapid growth, ensure that the number of the sporinite of later period abundance
Amount, as the rapidly depleting of nutrients, pH are increased in fermentation medium, the phase initially forms greatly Paenibacillus polymyxa after stabilization
The sporinite and mycoderm of amount.
Preferably, the condition of the actication of culture are as follows: cultivate 24~48h at 25~35 DEG C；It is further preferred that at 28~30 DEG C
Cultivate 36~48h.
Preferably, the condition of the seed culture are as follows: at 25~35 DEG C cultivate 12~for 24 hours；Revolving speed be 140~
200rpm；It is further preferred that cultivating 18-24h at 28~30 DEG C；Revolving speed is 150~180rpm.
Preferably, the condition of the fermented and cultured are as follows: cultivate 48~72h at 25~35 DEG C；Revolving speed be 250~
400rpm；It is further preferred that cultivating 48~72h at 28~30 DEG C；Revolving speed is 300~350rpm.
Paenibacillus polymyxa fermentation material made from above-mentioned preparation method, the how viscous class bud are utilized the present invention provides a kind of
The spore count of spore bacillus fermentation object is 1010-1011A/mL.
The present invention also provides a kind of sporinite micro-capsules, are calculated in mass percent, are made of following components:
100 parts of Paenibacillus polymyxa fermentation material；
1~10 part of polyethylene glycol；
1~20 part of soybean lecithin；
The Paenibacillus polymyxa fermentation material is as described above.
The present invention also provides a kind of sporinite microcapsule suspending agents, are calculated in mass percent, are made of following components:
The sporinite micro-capsule is as described above.
Further, the dispersing agent is at least one of polycarboxylate, organophosphorus ester and naphthalene sulfonate polymer.
Further, the emulsifier is polyester complexes, organophosphorus ester, EO/PO block polyether, castor oil polyoxy second
Alkene ether, in fatty alcohol polyoxyethylene ether, polyoxyethylene sorbitan fatty acid ester and sorbitan fatty acid ester at least
It is a kind of.
Further, the suspending agent is at least one of bentonite, white carbon black and aluminium-magnesium silicate.
The present invention also provides sporinite microcapsule suspending agents in preparation prevention and treatment rice sheath blight disease, false smut and bakanae disease of rice
Application in medicament.
Further, the amount of application of the sporinite microcapsule suspending agent is 30-300ml.
Compared with prior art, the invention has the following advantages:
(1) preparation method of the present invention is by selecting specific Paenibacillus polymyxa, changing seed culture medium and fermentation training
The component type and dosage for supporting base are fermented, and can be obtained the fermentation rich in mycoderm and Paenibacillus polymyxa sporinite and be produced
Object significantly improves the spore rate and mycoderm content of Paenibacillus polymyxa.
(2) sporinite micro-capsule provided by the invention and with sporinite micro-capsule prepare suspending agent have resistance of rainwater washing against and
Rice multiple diseases are all had good control efficiency, effectively can substitute or reduce chemical pesticide by the feature of lasting period length
It uses, and plant growth can be promoted, increase rice yield.
(3) Paenibacillus polymyxa zymotechnique provided by the invention can make the bacterial strain itself secretion generate a large amount of viscosity
Mycoderm can directly add using as natural biomembrane material, while with natural liposome biomembrane plant soybean lecithin
Work is formed by novel microcapsule through high molecular polymer PEG4000 modification, and processing technology is novel, green and pollution-free,
New thinking is provided for the preparation processing of biological pesticide.
(4) present invention successfully develops a kind of biomembrane itself secreted out of that can efficiently use and carrys out its viable spore of packing,
With environmentally protective biomaterial come further processing improvement forms novel microbial pesticide, the processing technology is in plant disease
New thinking is provided in the development effort of green prevention and control.
Detailed description of the invention
Fig. 1 be embodiment 1 in Paenibacillus polymyxa (ZJU-N) antagonism Rhizoctonia solani Kuhn (1), ustilaginoidea virens (2) and
The result figure of fusarium moniliforme (3)；
Wherein, A is control CK；B is Paenibacillus polymyxa (ZJU-N).
Fig. 2 is the installation for fermenting figure of Paenibacillus polymyxa in embodiment 2；
Fig. 3 is that the anti-rainwater of 3 miospore body microcapsule suspending agent of embodiment drenches map brushing；
Wherein, the spore liquid (right side) of microcapsule suspending agent it is not made including homemade sporinite microcapsule suspending agent (left side) and.
Specific embodiment
The invention will be further described combined with specific embodiments below, and what is be exemplified below is only specific implementation of the invention
Example, but protection scope of the present invention is not limited only to this.
Experimental method used in following embodiments is conventional method unless otherwise specified, material used, reagent
Deng being commercially available unless otherwise specified.
The indoor bioassay of 1 Paenibacillus polymyxa of embodiment
1, endophyte is separated from rice
From plant resources in Wenling, the rice bakanae disease of Jinhua Region, the withered morbidity field of line, the rice strain of picking health takes middle part up and down
Plant leaf diameter bar, is cut into the fritter of 5mm × 5mm, rinses 3 times, removes with 75% ethanol disinfection 1min or so, then with aqua sterilisa
Remaining disinfectant finally places it in and sucks extra moisture on sterile blotting paper, then puts into sterile mortar, is added
6mL BPS buffer is ground, and is taken 10,50,100 times of supernatant dilutions, is taken each 200 μ L of dilution to be coated on LB plate, verify simultaneously
Whether last time flushing water is coated on LB plate, examine tissue surface sterilization thorough, 30 DEG C of dark culture 1-2d, and records bacterium colony
Form, size and color.
The biological property of above-mentioned isolated Paenibacillus polymyxa are as follows: bacterium colony forms milky on NA culture medium
Bacterium colony, translucent, bacterium colony is smooth, mucus shape.Gram staining is the positive, and somatic cells are in the shape of a rod, mostly viscous under adverse environmental factor
Series bacillus can generate oval hypopus-gemma in expanding sporangium, and specification is 1.4-1.8 × 0.5-0.8 μm.
Above-mentioned isolated Paenibacillus polymyxa is subjected to culture presevation, preservation information is as follows: strain name: mostly viscous class
Bacillus (Paenibacillus polymyxa) ZJU-N；Depositary institution's title: China typical culture collection center；It protects
Hide unit address: Wuhan, China Wuhan University；Deposit number: CCTCC NO:M2018692；Preservation date: October 19 in 2018
Day.
2, inhibition of the indoor antagonistic experiment evaluation Paenibacillus polymyxa to rice sheath blight disease, false smut and bakanae disease of rice
Effect
Respectively about to rice banded sclerotial blight, false smut and the new activation culture of fusarium moniliforme plant weight of laboratory separation identification
3d, 10d and 5d or so, produce bacterium dish (diameter 0.5cm) on the active same circumference of strain growth, mycelia down and by its
It is transferred to PDA plate center, the filter paper being immersed in bacterium solution is taken out with sterilizing tweezers, is placed on from pathogen dish 2.5cm
It on plate, is compareed with sterile liquid filter paper, 4 repetitions are done in experiment, then culture dish are upside down in 25 DEG C of incubators, rice
The withered culture 4d of line, rice song culture 10d, bakanae disease culture 5d, taking-up see whether to be provided with apparent inhibiting effect.
As shown in Fig. 1 (A1 and B1), when the withered colony diameter of line for measuring CK control plate is 8.9cm, add Antagonistic Fungi
The withered bacterium of line in ZJU-N plate only long 2.2cm, inhibiting rate have reached 80%, illustrate Paenibacillus polymyxa ZJU-N to rice
Sheath blight fungus has apparent antagonistic effect.
As shown in Fig. 1 (A2 and B2), when the rice song colony diameter for measuring CK control plate is 3.6cm, add Antagonistic Fungi
Ustilaginoidea virens in ZJU-N plate is only 1.5cm, and inhibiting rate has reached 67%, illustrates Paenibacillus polymyxa ZJU-N to rice song
Germ has apparent antagonistic effect.
As shown in Fig. 1 (A3 and B3), when the bakanae disease of rice bacterium colony diameter for measuring CK control plate is 3.8cm, add short of money
Ustilaginoidea virens in antibacterial ZJU-N plate is 2.1cm, and inhibiting rate has reached 52%, illustrated Paenibacillus polymyxa ZJU-N to water
Rice Fusarium moniliforme Sheld has obvious antagonistic effect.
The zymotechnique of 2 Paenibacillus polymyxa of embodiment (ZJU-N)
The present embodiment fermentation culture medium used is shown in Table 1.
1 the present embodiment of table fermentation culture medium used summarizes
Equipment: the complete Zymolysis Equipment of Paenibacillus polymyxa (ZJU-N) liquid is purchased limited in Shanghai Chu Yi biotechnology
Company.
Zymotechnique process:
(1) actication of culture: one ring Paenibacillus polymyxa ZJU-N of picking will be inoculated in activation of crossing on solid plate
Plate be placed in 30 DEG C of incubators and cultivate 72h；
(2) seed culture: an independent clones of the Paenibacillus polymyxa of picking step (1) activation, which are inoculated in, to be equipped with
In the 250ml triangular flask of 100ml seed culture fluid, cultivated for 24 hours under conditions of temperature is 30 DEG C, revolving speed 160rpm；
(3) fermented and cultured: by cultured seed liquor with the 15L of 3% fermentation medium of the inoculum concentration access containing 13L
In fermentor, control fermentation jar temperature be 30 DEG C, the dissolved oxygen of revolving speed 300rpm, 80%-100%, after fermentation starts, often
It is sampled microscopy every 12h, until seeing the unified gemma of form (almost without thallus presence) under microscopy, then carries out gemma rate
Detection, observe viscous secretion biology mycoderm generation, finally determine fermentation terminate time and zymotechnique superiority and inferiority, finally
Obtain Paenibacillus polymyxa fermentation material.
Experiment is to optimize fermentation culture 13L, and 30 DEG C of temperature, the dissolved oxygen of revolving speed 300rpm, 80-100%, 3%
Inoculum concentration carry out the fermentation (such as Fig. 2) of 15L fermentor, determine under microscopy spore production rate 109A/mL, gemma rate up to 95% with
On, observe that a large amount of sticky mycoderms of secretion need 3-4 days.
The optimization of 3 zymotechnique of embodiment
The optimization that the present embodiment carries out fermentation medium: in basal medium (tryptone: 5g/L；Beef extract: 3g/
L；Yeast extract: 1g/L；NaCl:3g/L；Na2HPO4.12H2O:1.5g；Ca(NO3)2.4H2O:0.5g it is investigated on the basis of)
Several frequently seen carbon source (dosage of carbon source is same as Example 2) to Paenibacillus polymyxa viable count, produce gemma amount and mycoderm
Secrete the influence of situation.It is shown in Table 2.Remaining step is identical with embodiment 2.
Table 2: influence of the different carbon source to Paenibacillus polymyxa (ZJU-N) viable count, sporulation quantity and mycoderm secretory volume
Note: mycoderm secretory volume is more " +++ ", more " ++ ", less "+", no secretion "-"
By above-mentioned table 2 as it can be seen that potato and sucrose are more appropriate carbon sources, while after above-mentioned single factor experiment,
Further screening has been made to the concentration of carbon source, nitrogen source and inorganic salts, has obtained more appropriate ratio, i.e., shown in embodiment 2
Fermentation medium component and dosage, the result of viable count and gemma rate is shown in embodiment 2.
The preparation of the novel sporinite microcapsule suspending agent of 4 Paenibacillus polymyxa of embodiment and Field information 1, novel sporinite
The preparation of microcapsule suspending agent and anti-rain drop erosion measurement
(1) preparation of sporinite micro-capsule: taking solid-state high-purity soybean lecithin 10g, cholesterol dosage 5g, and chlorination is imitative after mixing
Impregnate dissolution, be evaporated under reduced pressure chloroform, lipid (i.e. the mixture of soybean lecithin and cholesterol) in bottle wall formed thin film after,
Paenibacillus polymyxa fermentation material (that is: the mixture of gemma and sticky mycoderm) prepared by 100ml embodiment 2 is added, then leads to
Rotating manipulation is received and distributed, multilamellar liposome is made, the PEG4000 that mass fraction is 2.5% is during which added, stirs evenly, after a period of time,
The sporinite biomembrane liposome (that is: sporinite micro-capsule) of PEG cladding can be obtained.
(2) preparation of sporinite microcapsule suspending agent: being calculated in mass percent, and takes sporinite micro-capsule 20% obtained, dispersion
Agent DS555 3%, Emulsifier O F-3463 12%, emulsifier 2,280 5%, suspending agent swelling organoclay 2%, mixing shearing, system
It is standby to obtain sporinite microcapsule suspending agent, it is used for field.
(3) resistance of rainwater washing against measures: spore is added dropwise in artificial membrane on glass slide, simulated rice plant surface wax layer respectively
Daughter microcapsule suspending agent (left side) and spore liquid (right, i.e., the how viscous class only obtained through the fermentation of embodiment 2 that microcapsule suspending agent is not made
Fermentation of bacillus object), Simulated rainwater uniformly washes away, and observes result (such as Fig. 3).
2, control efficiency and determination of yield of the field application sporinite microcapsule suspending agent to rice sheath blight disease
It is administered place: Jiaxing academy of agricultural sciences base Gu Tang time: Rise's boot period
Processing group 1: Paenibacillus polymyxa sporinite microcapsule suspending agent (the present embodiment first part preparation comprising mycoderm
Obtain), 300,000,000,000 CFU/ mus；
Processing group 2: the Paenibacillus polymyxa sporinite microcapsule suspending agent of sterile film, preparation method is: using except fermentation
The carbon source of culture medium is outside fructose, remaining content and the identical zymotechnique of embodiment 2 prepare fermentation material, then using implementation
The identical method of 4 first part of example prepares sporinite micro-capsule and suspending agent, and ensures sporinite quantity and 1 phase of processing group
Together；300000000000 CFU/ mus；
Processing group 3:24% thifluzamide SC, 20 milliliters/mu；
Processing group 4: it is not administered control.
Plot area 33m2, 4 repetitions.
Investigation method: every cell is sampled according to 5 points of diagonal line, every investigation 0.5m2, record total strain number, diseased plant number and disease
Series.
Control time: the 14th day after application.
Drug effect calculation method: it calculates as follows:
In formula: PT: control efficiency, unit are percentage (%)；CK1: disease index after blank district application；PT1: medicine
Disease index after the application of agent treatment region.
It surveys to produce and be carried out after rice is mature with species test, 6 points of every place's grab sample, every 0.25m2, add up to 1.5m2, investigate fringe
Number weighs and measures mass of 1000 kernel after drying (each repeating groups are average).
The comparison of 3 rice sheath blight disease field control effectiveness test result of table
Table 4 tests field and surveys production and species test result
As a result as shown in table 3, table 4, above-mentioned Paenibacillus polymyxa sporinite microcapsule suspending agent can effectively prevent rice
The generation of banded sclerotial blight, preventive effect are slightly below chemical agent thifluzamide, can be used for substituting or reducing the use of chemical pesticide, and slow down
The generation of drug resistance；From production and species test interpretation of result is surveyed, from microorganism formulation (that is: handling 1) is created than compareing volume increase 3.3%.
Claims (10)
1. a kind of preparation method of Paenibacillus polymyxa fermentation material characterized by comprising strain is successively activated,
Seed culture and fermented and cultured after being centrifuged, being concentrated, obtain Paenibacillus polymyxa fermentation material；The fermentation material be mycoderm and
The mixture of Paenibacillus polymyxa sporinite；
The strain is named as Paenibacillus polymyxa (Paenibacillus polymyxa) ZJU-N, deposit number are as follows:
CCTCC NO:M2018692, the deposit date is on October 19th, 2018；
The medium component of the seed culture are as follows: sucrose 10-15g/L, tryptone 5-10g/L, yeast extract 3-5g/L,
Beef extract 1.5-3g/L, NaCl 8-10g/L, pH 7.0-7.2；
The medium component of the fermented and cultured are as follows: potato 220-250g/L, tryptone 5-8g/L, sucrose 10-15g/L,
Yeast extract 1-5g/L, beef extract 0.5-1g/L, NaCl 1-5g/L, Ca (NO3)2·4H2O 0.5-2g/L,
Na2HPO4.12H2O 1.5-3g/L, pH7.0-7.2.
2. the preparation method of Paenibacillus polymyxa fermentation material as described in claim 1, which is characterized in that the actication of culture
Condition are as follows: at 25~35 DEG C cultivate 24~48h.
3. the preparation method of Paenibacillus polymyxa fermentation material as described in claim 1, which is characterized in that the seed culture
Condition are as follows: at 25~35 DEG C cultivate 12~for 24 hours；Revolving speed is 140~200rpm.
4. the preparation method of Paenibacillus polymyxa fermentation material as described in claim 1, which is characterized in that the fermented and cultured
Condition are as follows: at 25~35 DEG C cultivate 48~72h；Revolving speed is 250~400rpm.
5. it is a kind of using Paenibacillus polymyxa fermentation material made from the described in any item preparation methods of Claims 1 to 44, it is special
Sign is that the spore count of the Paenibacillus polymyxa fermentation material is 1010-1011A/mL.
6. a kind of sporinite micro-capsule, which is characterized in that in terms of weight fraction, be made of following components:
100 parts of Paenibacillus polymyxa fermentation material；
1~10 part of polyethylene glycol；
1~20 part of soybean lecithin；
The Paenibacillus polymyxa fermentation material is as claimed in claim 5.
7. a kind of sporinite microcapsule suspending agent, which is characterized in that be calculated in mass percent, be made of following components:
Remaining is water；
The sporinite micro-capsule is as claimed in claim 6.
8. sporinite microcapsule suspending agent as claimed in claim 7, which is characterized in that the dispersing agent is polycarboxylate, organic
At least one of phosphate and naphthalene sulfonate polymer；
The emulsifier is polyester complexes, organophosphorus ester, EO/PO block polyether, castor oil polyoxyethylene ether, poly alkyl alcohol
At least one of ethylene oxide ether, polyoxyethylene sorbitan fatty acid ester and sorbitan fatty acid ester；
The suspending agent is at least one of bentonite, white carbon black and aluminium-magnesium silicate.
9. if the described in any item sporinite microcapsule suspending agents of claim 6~8 are in prevention and treatment rice sheath blight disease, false smut and rice
Application in bakanae disease medicament.
10. as claimed in claim 9 application, which is characterized in that the amount of application of the sporinite microcapsule suspending agent be 30~
300ml。
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